Feasibility study on a perceived fatigue prediction dependent power control for an electrically assisted bicycle.

Several types of electric motor assists have been developed, as a result, it is important to control muscular fatigue on-site in terms of health promotion and motor rehabilitation. Predicting the perceived fatigue by several biosignal-related variables with the multiple regression model and polynomial approximation, we try to propose a self control design for the electrically assisted bicycle (EAB). We also determine the meaningful muscles during pedaling by muscle synergies in relation to the motion maturity. In field experiments, prediction of ongoing perceived physical fatigue could have the potential of suitable control of EAB.

Identification of the clonal complexes of Staphylococcus aureus strains by determination of the conservation patterns of small genomic islets.

AIMS: To investigate the clonality of Staphylococcus aureus isolates, it is important to identify their clonal complexes (CCs) with multilocus sequence typing (MLST). However, it is expensive to carry out MLST analyses for many isolates. The aim of this study, therefore, was to develop a cost-effective method to identify CCs by determining the conservation pattern of 'small genomic islets' (SGIs). SGIs are nonconserved regions between strains and have single or multiple open-reading frames (ORFs). METHODS AND RESULTS: The whole-genome sequences of nine strains were compared in order to select 16 SGIs. The conservation patterns of the 16 SGIs (islet patterns) were investigated in 136 S. aureus isolates, which were classified into 21 CCs. The islet patterns (IPs) exhibited a one-to-one correspondence with the CCs, except for isolates belonging to CC1, CC5 and CC8. The IPs typical of strains belonging to CC1, CC5 and CC8 differed between those of sequence type 1 (ST1) and ST188 (CC1), ST5 and ST6 (CC5) and ST8 and ST239 (CC8). SIGNIFICANCE AND IMPACT OF THE STUDY: The CCs of many isolates can be identified in an easy and inexpensive manner by detecting these 16 SGIs. Emergent clones, particularly methicillin-resistant ones, can be identified by examining numerous islets by IP analysis.

Improving the thermal stability of lactate oxidase by directed evolution.

Lactate oxidase is used in biosensors to measure the concentration of lactate in the blood and other body fluids. Increasing the thermostability of lactate oxidase can significantly prolong the lifetime of these biosensors. We have previously obtained a variant of lactate oxidase from Aerococcus viridans with two mutations (E160G/V198I) that is significantly more thermostable than the wild-type enzyme. Here we have attempted to further improve the thermostability of E160G/V198I lactate oxidase using directed evolution. We made a mutant lactate oxidase gene library by applying error-prone PCR and DNA shuffling, and screened for thermostable mutant lactate oxidase using a plate-based assay. After three rounds of screening we obtained a thermostable mutant lactate oxidase, which has six mutations (E160G/V198I/G36S/T103S/A232S/F277Y). The half-life of this lactate oxidase at 70 degrees C was about 2 times that of E160G/V198I and about 36 times that of the wild-type enzyme. The amino acid mutation process suggests that the combined neutral mutations are important in protein evolution.

The measles virus hemagglutinin downregulates the cellular receptor SLAM (CD150).

Signaling lymphocyte activation molecule (SLAM), a cellular receptor for measles virus, was downregulated from the surface of cells infected with either the Edmonston or wild-type KA strain of measles virus. Transfection of the expression plasmid encoding the Edmonston or KA hemagglutinin, but not the fusion protein, induced downregulation of SLAM in not only cells expressing the envelope protein on the surface, but those not expressing it. After cocultivation with cells expressing the hemagglutinin, SLAM-expressing cells also exhibited downregulation of SLAM. Thus, the measles virus hemagglutinin can induce downregulation of SLAM in cells either expressing or coming in contact with it.

A case of spindle cell carcinoma of the breast--long survival achieved by multiple surgical treatment.

Spindle cell carcinoma of the breast was formerly called carcinosarcoma, and is relatively rare. We report a case of spindle cell carcinoma of the breast. The patient was treated with multiple surgeries and achieved long-term survival. The patient was a 52-year-old woman, in whom small induration developed at the areola of the nipple of the right breast. The lesion was resected, and benign tumor was diagnosed pathologically. Four years later, she had recurrence at the scar, and a typical mastectomy was performed. A tumor developed again 5 years later; the lesional focus was at the scar of the right chest wall and invasion of the ribs and the sternum was noted. The sternum and the right costal cartilage of ribs 3-9 were dissected together. The right chest wall was reconstructed and adjuvant radiation therapy performed. Four years after this operation, tumor recurred near the scar and chest wall resection including part of the pericardial cavity and the left lung was performed. However, 6 months later, invasion of the mediastinum, heart and lung were noted. The patient died 16 years after the first surgery. Dermatofibrosarcoma protuberance of the breast was diagnosed at the second operation. However, the diagnosis was changed to spindle cell carcinoma of the breast following immunohistochemical studies. Spindle cell carcinoma of the breast is rare, and definitive histopathological diagnosis is often difficult. When spindle cell carcinoma is suspected, comprehensive diagnostic studies including immunohistochemical examinations should be performed. Even in case with multiple recurrences correctly performed operations may contribute to prolongation of survival.

Position of immobilization after dislocation of the glenohumeral joint. A study with use of magnetic resonance imaging.

BACKGROUND: Glenohumeral dislocations often recur, probably because a Bankart lesion does not heal sufficiently during the period of immobilization. Using magnetic resonance imaging, we assessed the position of the Bankart lesion, with the arm in internal and external rotation, in shoulders that had had a dislocation. METHODS: Coaptation of a Bankart lesion was examined with use of magnetic resonance imaging, with the arm held at the side of the trunk and positioned first in internal rotation (mean, 29 degrees) and then in external rotation (mean, 35 degrees), in nineteen shoulders. Six shoulders (six patients) had had an initial anterior dislocation, and thirteen shoulders (twelve patients) had had recurrent anterior dislocation. Fast-spin-echo T2-weighted axial images were made when the dislocation had occurred less than two weeks earlier, and spin-echo T1-weighted axial images after intra-articular injection of gadolinium-diethylenetriamine pentaacetic acid were made when the dislocation had occurred more than two weeks earlier. Separation and displacement of the anteroinferior portion of the labrum from the glenoid rim were measured on the axial images, and coaptation of the anterior part of the capsule to the glenoid neck was assessed by measurement of the detached area, opening angle, and detached length. RESULTS: Separation and displacement of the labrum were both significantly less (p = 0.0047 and p = 0.0017, respectively) when the arm was in external rotation than when it was in internal rotation. The detached area and the opening angle of the anteroinferior portion of the capsule were both significantly smaller (p = 0.0003 and p < 0.0001, respectively), and the detached length was significantly shorter (p < 0.0001) with the arm in external rotation. CONCLUSION: Immobilization of the arm in external rotation better approximates the Bankart lesion to the glenoid neck than does the conventional position of internal rotation.

Measles viruses on throat swabs from measles patients use signaling lymphocytic activation molecule (CDw150) but not CD46 as a cellular receptor.

Both CD46 and signaling lymphocytic activation molecule (SLAM) have been shown to act as cellular receptors for measles virus (MV). The viruses on throat swabs from nine patients with measles in Japan were titrated on Vero cells stably expressing human SLAM. Samples from all but two patients produced numerous plaques on SLAM-expressing Vero cells, whereas none produced any plaques on Vero cells endogenously expressing CD46. The Edmonston strain of MV, which can use either CD46 or SLAM as a receptor, produced comparable titers on these two types of cells. The results strongly suggest that the viruses in the bodies of measles patients use SLAM but probably not CD46 as a cellular receptor.

V domain of human SLAM (CDw150) is essential for its function as a measles virus receptor.

Human signaling lymphocytic activation molecule (SLAM; also known as CDw150) has been shown to be a cellular receptor for measles virus (MV). Chinese hamster ovary cells transfected with a mouse SLAM cDNA were not susceptible to MV and the vesicular stomatitis virus pseudotype bearing MV envelope proteins alone, indicating that mouse SLAM cannot act as an MV receptor. To determine the functional domain of the receptor, we tested the abilities of several chimeric SLAM proteins to function as MV receptors. The ectodomain of SLAM comprises the two immunoglobulin superfamily domains (V and C2). Various chimeric transmembrane proteins possessing the V domain of human SLAM were able to act as MV receptors, whereas a chimera consisting of human SLAM containing the mouse V domain instead of the human V domain no longer acted as a receptor. To examine the interaction between SLAM and MV envelope proteins, recombinant soluble forms of SLAM were produced. The soluble molecules possessing the V domain of human SLAM were shown to bind to cells expressing the MV hemagglutinin (H) protein but not to cells expressing the MV fusion protein or irrelevant envelope proteins. These results indicate that the V domain of human SLAM is necessary and sufficient to interact with the MV H protein and allow MV entry.

Latent herpes simplex virus-1 infection in SCID mice transferred with immune CD4+T cells: a new model for latency.

In C.B-17 severe combined immunodeficiency (SCID) mice, corneal challenge with herpes simplex virus-1 (HSV-1) KOS strain usually leads to fatal encephalitis. With the transfer of T cells from immunized BALB/c mice, these SCID mice developed a latent HSV-1 infection. In order to determine the responsible T cell subset, fractionated immune T cells were transferred. Those SCID mice transferred with immune CD4+T cell-enriched fraction developed latent HSV-1 infection in their trigeminal ganglia. Their splenocytes had an increased percentage of CD4+T cells and showed a proliferative response against HSV-1. The transfer of CD8+T cells increased survival in the acute infection, but their engraftment seemed less needed for latency than that of CD4+T cells. Mice that received immune serum survived without developing latent HSV-1 infection. Some latently infected SCID mice had anti-HSV antibodies while others did not, indicating that the engraftment of antibody-producing B cells was not required for latency. Thus, immune CD4+T cells were important for the survival of SCID mice with latent HSV-1 infection. This animal model should be useful for investigation of latency/reactivation of HSV-1.

Virus entry is a major determinant of cell tropism of Edmonston and wild-type strains of measles virus as revealed by vesicular stomatitis virus pseudotypes bearing their envelope proteins.

The Edmonston strain of measles virus (MV) that utilizes the human CD46 as the cellular receptor produced cytopathic effects (CPE) in all of the primate cell lines examined. In contrast, the wild-type MV strains isolated in a marmoset B-cell line B95a (the KA and Ichinose strains) replicated and produced CPE in some but not all of the primate lymphoid cell lines. To determine the mechanism underlying this difference in cell tropism, we used a recently developed recombinant vesicular stomatitis virus (VSV) containing as a reporter the green fluorescent protein gene in lieu of the VSV G protein gene (VSVDeltaG*). MV glycoproteins were efficiently incorporated into VSVDeltaG*, producing the VSV pseudotypes. VSVDeltaG* complemented with VSV G protein efficiently infected all of the cell lines tested. The VSV pseudotype bearing the Edmonston hemagglutinin (H) and fusion (F) protein (VSVDeltaG*-EdHF) infected all cell lines in which the Edmonston strain caused CPE, including the rodent cell lines to which the human CD46 gene was stably transfected. The pseudotype bearing the wild-type KA H protein and Edmonston F protein (VSVDeltaG*-KAHF) infected all lymphoid cell lines in which the wild-type MV strains caused CPE as efficiently as VSVDeltaG*-EdHF, but it did not infect any of the cell lines resistant to infection with the KA strain. The results indicate that the difference in cell tropism between these MV strains was largely determined by virus entry, in which the H proteins of respective MV strains play a decisive role.

Magnesium(II) and zinc(II)-protoporphyrin IX's stabilize the lowest oxygen affinity state of human hemoglobin even more strongly than deoxyheme.

Studies of oxygen equilibrium properties of Mg(II)-Fe(II) and Zn(II)-Fe(II) hybrid hemoglobins (i.e. alpha2(Fe)beta2(M) and alpha2(M)beta2(Fe); M=Mg(II), Zn(II) (neither of these closed-shell metal ions binds oxygen or carbon monoxide)) are reported along with the X-ray crystal structures of alpha2(Fe)beta2(Mg) with and without CO bound. We found that Mg(II)-Fe(II) hybrids resemble Zn(II)-Fe(II) hybrids very closely in oxygen equilibrium properties. The Fe(II)-subunits in these hybrids bind oxygen with very low affinities, and the effect of allosteric effectors, such as proton and/or inositol hexaphosphate, is relatively small. We also found a striking similarity in spectrophotometric properties between Mg(II)-Fe(II) and Zn(II)-Fe(II) hybrids, particularly, the large spectral changes that occur specifically in the metal-containing beta subunits upon the R-T transition of the hybrids. In crystals, both alpha2(Fe)beta2(Mg) and alpha2(Fe-CO)beta2(Mg) adopt the quaternary structure of deoxyhemoglobin. These results, combined with the re-evaluation of the oxygen equilibrium properties of normal hemoglobin, low-affinity mutants, and metal substituted hybrids, point to a general tendency of human hemoglobin that when the association equilibrium constant of hemoglobin for the first binding oxygen molecule (K1) approaches 0.004 mmHg(-1), the cooperativity as well as the effect of allosteric effectors is virtually abolished. This is indicative of the existence of a distinct thermodynamic state which determines the lowest oxygen affinity of human hemoglobin. Moreover, excellent agreement between the reported oxygen affinity of deoxyhemoglobin in crystals and the lowest affinity in solution leads us to propose that the classical T structure of deoxyhemoglobin in the crystals represents the lowest affinity state in solution. We also survey the oxygen equilibrium properties of various metal-substituted hybrid hemoglobins studied over the past 20 years in our laboratory. The bulk of these data are consistent with the Perutz's trigger mechanism, in that the affinity of a metal hybrid is determined by the ionic radius of the metal, and also by the steric effect of the distal ligand, if present. However, there remains a fundamental contradiction among the oxygen equilibrium properties of the beta substituted hybrid hemoglobins.

Amino acid substitutions at position 481 differently affect the ability of the measles virus hemagglutinin to induce cell fusion in monkey and marmoset cells co-expressing the fusion protein.

The hemagglutinin (H) protein of the measles virus (MV) Edmonston strain induced cell fusion in Cos (monkey) and B95a (marmoset) cells, when co-expressed with the fusion (F) protein, whereas the H protein of the wild-type KA strain induced fusion in B95a cells, but not in Cos cells. Asparagine residue at position 481 of the KA H protein was replaced by various amino acids through site-directed mutagenesis. Substitution with tyrosine, which was found at position 481 of the Edmonston H protein, enabled the mutant KA H protein (N481Y) to induce cell fusion in Cos cells co-expressing the F protein, which could be completely blocked by anti-CD46 antibody. This mutant, however, did not cause CD46 downregulation, unlike the Edmonston H protein. The other H protein mutants (N481S, N481T, N481D, N481H, N481F) did not produce syncytia in Cos cells. On the other hand, all of the mutants retained the ability to induce cell fusion in B95a cells. Thus, while tyrosine at position 481 was indispensable for the MV H protein's interaction with CD46, the residue at this position does not appear to be critically involved in the interaction with the receptor for wild-type strains present on B95a cells.

Enhanced IFN-gamma production in vitro by CD8+ T cells in hemophiliacs with AIDS as demonstrated on the single-cell level.

We examined T cells from HIV-infected hemophiliacs under antiviral therapy, on the single-cell level, for cytokine production in vitro in response to stimulation. The percentage of IL-2-producing cells was markedly decreased among both CD3+CD8- and CD3+CD8+ cells, while the proportion of IFN-gamma-producing cells was preserved among CD3+CD8- cells and increased among CD3+CD8+ cells in HIV+ subjects, compared with HIV-uninfected controls. The increase in IFN-gamma-producing CD8+ cells was accounted for by the expansion of CD8+CD28- cells among total CD8+ T cells and by the higher percentage of IFN-gamma-expressing cells among both CD8+CD28+ and CD8+CD28- cells in HIV+ individuals, compared with controls. The enhanced IFN-gamma production in CD8+ cells from individuals in the advanced phase of HIV infection might implicate the host's response to chronic viral infection or senescence of host CD8+ cells.

Humeral attachment of the supraspinatus and infraspinatus tendons: an anatomic study.

We investigated the anatomic relationship of the supraspinatus (SSP) and infraspinatus (ISP) tendons to the three facets of the greater tuberosity. After removing the superficial layer of the cuff to expose the tendon fibers in 10 embalmed shoulders, the cuff tendon attachment to the facets was examined, and the location of attachment was measured in reference to (1) the anterior margin of the greater tuberosity and (2) the superior margin of the sulcus (anatomic neck without cartilage). The SSP tendon attached to the superior facet and the superior half of the middle facet. The ISP tendon attached to the entire middle facet, covering a portion of the SSP tendon. Thus, the anterior half of the superior cuff tendon (12.6 +/- 1.1 mm) was composed of only the SSP tendon, whereas the posterior half (9.8 +/- 3.2 mm) was composed of both the SSP and ISP tendons. The sulcus was located not at the SSP-ISP interval but slightly posterior to the posterior margin of the SSP tendon (4.3 +/- 2.4 mm). We conclude that (1) there is an overlap between the SSP and ISP tendons identifiable by the facets or the distance from the anterior greater tuberosity and (2) the sulcus is located slightly posterior to the posterior margin of the SSP tendon.

Plasma levels of cyclic GMP, immune parameters and depressive status during interferon therapy. A prospective study in Japan.

In this report, we investigated the relationship between depressive symptoms and plasma interferon (IFN)-alpha-like immunoreactivity, cyclic GMP (cGMP) and soluble interleukin-2 receptor (sIL-2R) levels during IFN therapy. An altered mood state was observed in 5 of 26 patients. IFN-alpha-like immunoreactivity in the depressed group tended to be elevated. cGMP levels of depressed patients were significantly greater than those of control subjects before and 6 weeks after IFN therapy. However, sIL-2R levels were not different between the two groups. These results suggest that a number of patients suffered from depression during IFN therapy and that patients had greater concentrations of cGMP levels.

Isokinetic strength after tears of the supraspinatus tendon.

We measured the isokinetic strength of abduction, adduction, internal rotation, and external rotation in ten patients with full-thickness tears of the supraspinatus and ten with partial-thickness tears. The measurements were repeated after intra-articular or intrabursal injection of local anaesthetic. Pain blocks produced significant increases in strength in both full and partial-thickness tears. After the block, the strength in full-thickness tears compared with the opposite side was 67% to 81% in abduction and 67% to 78% in external rotation, both significantly smaller than those on the uninvolved side (p = 0.0064, p = 0.0170). In partial-thickness tears the strength after the block ranged from 82% to 111%, with no significant differences between the involved and uninvolved sides. The decreases in strength of 19% to 33% in abduction and 22% to 33% in external rotation after full-thickness tears appear to represent the contribution of supraspinatus to the strength of the shoulder.

Pathogenicity of glycoprotein C-deficient herpes simplex virus 1 strain TN-1 which encodes truncated glycoprotein C.

A clinical isolate of herpes simplex virus 1 (TN-1) from a stromal keratitis patient was found to be defective in the glycoprotein C (gC) gene (UL44), thus resulting in the production of truncated gC upon infection. To study the pathogenetic role of truncated gC, we prepared a recombinant LTN-8 derived from TN-1 with deletions of the 1.5 kilobase pairs of the gC gene including the initiation codon. A penetration assay revealed LTN-8 to be less efficient in its penetration ability than TN-1, the laboratory strain KOS and RTN-1-20-3, a recombinant derived from TN-1 with the KOS gC gene. The penetration of LTN-8 was facilitated by the addition of TN-1-infected culture medium. TN-1 virus preparations had no hemagglutinating activity. However, the animals infected with TN-1 did develop hemagglutination inhibition (HI) antibodies. The LTN-8-infected animals did not develop HI antibodies. The pathogenicity in BALB/c mice following either corneal, intraperitoneal or intracerebral inoculation did not significantly differ among TN-1, RTN-1-20-3 or LTN-8. Our results indicate that truncated gC was sufficient for the induction of HI antibodies and was also able to facilitate penetration in vitro. Although truncated gC might be a virulence factor acting as a decoy, both truncated gC and intact gC had little effect on the outcome following intracerebral, intraperitoneal or corneal inoculation.

Suppression of infectious virus spread and corneal opacification by the combined use of recombinant interferon beta and interleukin-10 following corneal infection with herpes simplex virus-1 in mice.

The effects of Interleukin-10 (IL-10) and recombinant murine interferon-beta (rMuIFN-beta) on experimental corneal herpes simplex virus 1 (HSV-1) inoculation in BALB/c mice were examined. The mice were inoculated with the HSV-1 strain KOS at their corneas after abrasion. IL-10 was then administered topically once a day for 10 days beginning 2 days post inoculation, while rMuIFN-beta was administered once a day for 10 days beginning 1 day post inoculation. The local viral growth in the inoculated eyes and trigeminal ganglia was reduced in the rMuIFN-beta-treated mice but not in the IL-10-treated mice. In the mice treated with both rMuIFN-beta and IL-10, the degree of both the local viral growth and corneal opacification decreased. The establishment of HSV-1 latency in the trigeminal ganglia was partially prevented by rMuIFN-beta treatment but not by IL-10 treatment. The combined use of the cytokines resulted in both the suppression of viral spread and the prevention of corneal inflammation induced by HSV-1 infection.

Tactile-audio diagram for blind persons.

We propose a tactile-audio diagram, represented by a tactile pattern and linguistic information, as a new media for the blind. We have developed a vision substitution system which enables blind persons to read and write a diagram in the form of a tactile-audio diagram. The system consists of a personal computer, a tactile display with digitizer, an auditory information input/output device, and a command keyboard. The features of the system are: 1) the use of multimodality, i.e., tactile and auditory senses are used for perceiving spatial and linguistic information and 2) two-way communication, i.e., the system enables a blind person to read and write a diagram. This paper presents a tactile-audio diagram and an implemented prototype system based on this theory, along with a discussion and methods for enhanced representations.

Successful foscarnet therapy for mucocutaneous infection with herpes simplex virus in a recipient after unrelated bone marrow transplantation.

A 36-year-old Japanese man who received an unrelated bone marrow transplant (BMT) developed severe mucocutaneous infection with herpes simplex virus (HSV) type 1 during oral acyclovir prophylaxis. The lesions progressed despite treatment with intravenous acyclovir and vidarabine. The HSV isolates were sensitive acyclovir, vidarabine and foscarnet in vitro, but peripheral CD3- or CD19-positive cells were barely detectable even 4 months after transplant. A 12-day course of treatment with foscarnet led to a rapid improvement. Foscarnet therapy should be considered for all severe HSV infections following BMT, regardless of whether or not the HSV isolates are sensitive to acyclovir.