Gait posture estimation using wearable acceleration and gyro sensors

A method for gait analysis using wearable acceleration sensors and gyro sensors is proposed in this work. The volunteers wore sensor units that included a tri-axis acceleration sensor and three single axis gyro sensors. The angular velocity data measured by the gyro sensors were used to estimate the translational acceleration in the gait analysis. The translational acceleration was then subtracted from the acceleration sensor measurements to obtain the gravitational acceleration, giving the orientation of the lower limb segments. Segment orientation along with body measurements were used to obtain the positions of hip, knee, and ankle joints to create stick figure models of the volunteers. This method can measure the three-dimensional positions of joint centers of the hip, knee, and ankle during movement. Experiments were carried out on the normal gait of three healthy volunteers. As a result, the flexion–extension (F–E) and the adduction–abduction (A–A) joint angles of the hips and the flexion–extension (F–E) joint angles of the knees were calculated and compared with a camera motion capture system. The correlation coefficients were above 0.88 for the hip F–E, higher than 0.72 for the hip A–A, better than 0.92 for the knee F–E. A moving stick figure model of each volunteer was created to visually confirm the walking posture. Further, the knee and ankle joint trajectories in the horizontal plane showed that the left and right legs were bilaterally symmetric.     

Molecular and genetic characterization of the <Emphasis Type="Italic">S</Emphasis> locus in <Emphasis Type="Italic">Hordeum bulbosum</Emphasis> L., a wild self-incompatible species related to cultivated barley

Gametophytic self-incompatibility (GSI) in the grasses is controlled by a distinct two-locus genetic system governed by the multiallelic loci S and Z. We have employed diploid Hordeum bulbosum as a model species for identifying the self-incompatibility (SI) genes and for elucidating the molecular mechanisms of the two-locus SI system in the grasses. In this study, we attempted to identify S haplotype-specific cDNAs expressed in pistils and anthers at the flowering stage in H. bulbosum, using the AFLP-based mRNA fingerprinting (AMF, also called cDNA-AFLP) technique. We used the AMF-derived DNA clones as markers for fine mapping of the S locus, and found that the locus resided in a chromosomal region displaying remarkable suppression of recombination, encompassing a large physical region. Furthermore, we identified three AMF-derived markers displaying complete linkage to the S locus, although they showed no significant homology with genes of known functions. Two of these markers showed expression patterns that were specific to the reproductive organs (pistil or anther), suggesting that they could be potential candidates for the S gene.     

Inhibitory effect of intracerebroventricularly-administered [D-Arg(2), beta-Ala(4)]-dermorphin (1-4) on gastrointestinal transit

The inhibitory effect of intracerebroventricularly-administered [D-Arg(2), beta-Ala(4)]-dermorphin (1-4) (TAPA), a highly selective mu(1)-opioid receptor agonist, on mouse gastrointestinal transit was compared with that of morphine and [D-Ala(2), N-methyl-Phe(4), Gly(5)-ol]-enkephalin (DAMGO). When administered intracerebroventricularly 5 min before the oral injection of charcoal meal, TAPA (10-100 pmol), morphine (0.25-4 nmol), and DAMGO (20-80 pmol) dose-dependently inhibited gastrointestinal transit of charcoal. The inhibitory effect of each mu-opioid receptor agonist was completely antagonized by naloxone, a nonselective opioid receptor antagonist. The inhibitory effects of morphine and DAMGO were significantly antagonized by both beta-funaltrexamine, a selective mu-opioid receptor antagonist, and naloxonazine, a selective mu(1)-opioid receptor antagonist. In contrast, the inhibitory effect of TAPA was not affected at all by beta-funaltrexamine, naloxonazine, nor-binaltorphimine (a selective kappa-opioid receptor antagonist), or naltrindole (a selective delta-opioid receptor antagonist). These results suggest that the inhibitory effect of TAPA on gastrointestinal transit may be mediated through an opioid receptor mechanism different from that of morphine and DAMGO.     

Mutational analyses of restriction endonuclease-HindIII mutant E86K with higher activity and altered specificity

We have performed mutational analyses of restriction endonuclease HindIII in order to identify the amino acid residues responsible for enzyme activity. Four of the seven HindIII mutants, which had His-tag sequences at the N-termini, were expressed in Escherichia coli, and purified to homogeneity. The His-tag sequence did not affect enzyme activity, whereas it hindered binding of the DNA probe in gel retardation assays. A mutant E86K in which Lys was substituted for Glu at residue 86 exhibited high endonuclease activity. Gel retardation assays showed high affinity of this mutant to the DNA probe. Surprisingly, in the presence of a transition metal, Mo(2+) or Mn(2+), the E86K mutant cleaved substrate DNA at a site other than HindIII. Substitution of Glu for Val at residue 106 (V106E), and Asn for Lys at residue 125 (K125N) resulted in a decrease in both endonucleolytic and DNA binding activities of the enzyme. Furthermore, substitution of Leu for Asp at residue 108 (D108L) abolished both HindIII endonuclease and DNA binding activities. CD spectra of the wild type and the two mutants, E86K and D108L, were similar to each other, suggesting that there was little change in conformation as a result of the mutations. These results account for the notion that Asp108 could be directly involved in HindIII catalytic function, and that the substitution at residue 86 may bring about new interactions between DNA and cations.     

Nuclear localization of Japanese encephalitis virus core protein enhances viral replication

Japanese encephalitis virus (JEV) core protein was detected in both the nucleoli and cytoplasm of mammalian and insect cell lines infected with JEV or transfected with the expression plasmid of the core protein. Mutation analysis revealed that Gly(42) and Pro(43) in the core protein are essential for the nuclear and nucleolar localization. A mutant M4243 virus in which both Gly(42) and Pro(43) were replaced by Ala was recovered by plasmid-based reverse genetics. In C6/36 mosquito cells, the M4243 virus exhibited RNA replication and protein synthesis comparable to wild-type JEV, whereas propagation in Vero cells was impaired. The mutant core protein was detected in the cytoplasm but not in the nucleus of either C6/36 or Vero cell lines infected with the M4243 virus. The impaired propagation of M4243 in mammalian cells was recovered by the expression of wild-type core protein in trans but not by that of the mutant core protein. Although M4243 mutant virus exhibited a high level of neurovirulence comparable to wild-type JEV in spite of the approximately 100-fold-lower viral propagation after intracerebral inoculation to 3-week-old mice of strain Jcl:ICR, no virus was recovered from the brain after intraperitoneal inoculation of the mutant. These results indicate that nuclear localization of JEV core protein plays crucial roles not only in the replication in mammalian cells in vitro but also in the pathogenesis of encephalitis induced by JEV in vivo.     

Suppressive effect of nantenine,isolated from Nandina domestica Thunberg,on the 5-hydroxy-L-tryptophan plus clorgyline-induced head-twitch response in mice

We investigated the effects of nantenine (9,10-Methylenedioxy-1,2 dimethoxyaporphine), a major alkaloid isolated from the fruit of Nandina domestica Thunb (Berberidaceae), on the 5-HT2A receptor-mediated head-twitch response (HTR) in mice. Intraperitoneal (i.p.) injection of nantenine (13.3, 20 and 30 mg/kg) as well as the 5-HT2A receptor antagonist ketanserin (0.0625, 0.25 and 1 mg/kg) inhibited the 5-hydroxy-L-tryptophan (l-5-HTP; 75 mg/kg, i.p.) plus monoamine oxidase inhibitor, clorgyline (1 mg/kg, i.p.)-induced HTR in a dose-dependent manner. In contrast, neither l-5-HTP plus clorgyline nor 5-HT1A agonist, 8-hydroxy-2-(di-n-propylamino)tetraline (8-OH-DPAT; 5 microg/mouse, i.c.v.)-induced head weaving was affected by nantenine or ketanserin. Furthermore, neither nantenine (up to 30 mg/kg) nor ketanserin (up to 1 mg/kg) affect on the locomotor activity. In the receptor binding studies, nantenine showed affinity to the 5-HT2A receptors (Ki = 0.4 microM), while it had less affinity toward alpha1-adrenergic (Ki = 2.1 microM) and D2-dopaminergic (Ki = 1.7 microM) receptors of the mouse brain. These results suggest that nantenine inhibits l-5-HTP plus clorgyline-induced head- twitch response by blocking 5-HT2A receptors in the central nervous system.     

Selection and characterization of a nickel-tolerant cell line from tobacco (Nicotiana tabacum cv. bright yellow-2) suspension culture

To investigate the cellular/molecular mechanisms of nickel (Ni) tolerance in high-biomass-producing plants, we selected a Ni-tolerant (NIT) cell line from tobacco ( Nicotiana tabacum L. cv. bright yellow-2) suspension culture. Examination of response to various abiotic stresses showed that the NIT cells acquired remarkable tolerance to excess Ni, Cu, and Al but not to the other stresses. That the NIT cells accumulated approximately 2 m M Ni under the 700 µ M Ni condition suggests that the NIT cells can hyperaccumulate Ni and possess internal detoxification mechanisms for Ni. Two different sizes of putative β- d -xylosidase were constitutively secreted by the NIT cells, and the NIT cells showed greater elongation than the wild-type (WT) cells. Oxalate, citrate, 2-oxoglutarate, and glutamate contents were constitutively two- or three-fold higher in the NIT cells than in the WT cells, and histidine content was increased by up to three-fold in the NIT cells exposed to 700 µ M Ni compared to those in the WT cells. Newport green DCF diacetate (NPG), a Ni-specific fluorescence indicator, enabled the visualization of the subcellular localization of Ni in both WT and NIT cells. The NPG fluorescence was localized mainly in the vacuoles regardless of the WT or NIT cells under excess Ni conditions. The transport of Ni-organic acids and/or Ni-histidine complex into the vacuoles might be an important mechanism contributing to the high Ni tolerance of and the hyperaccumulation of Ni by the NIT cells.     

Secretion of acid phosphatase by the roots of crop plants under phosphorus-deficient conditions and some properties of the enzyme secreted by lupin roots

Nine crop species were grown in P-sufficient and P-deficient nutrient solutions. The activity of acid phosphatase secreted by the roots increased under P-deficient conditions in all the species examined. That of lupin increased most remarkably. The properties of the enzyme secreted by the roots of lupin was investigated. Many isozymes existed in the roots and the leaves, but only one of them was secreted into the rhizosphere in a large amount. The molecular weight of the purified enzyme secreted was estimated to be 72 KD by SDS-PAGE and 140 KD by gel filtration; it was assumed to be a homo-dimer. The iso-electric point of the enzyme was 4.7 and the pH for optimum activity 4.3. When the enzyme was mixed with aqueous solution extracted from a P-deficient soil, its activity declined to 55% of its original activity after 14 days and to 9% after 21 days.     

Biphasic effects of yohimbine on the ejaculatory response in the dog

The effects of various doses (0.01-1.00 mg/kg) of yohimbine, an alpha-2 adrenoceptor antagonist, on the erectile and ejaculatory response elicited by manual penile stimulation were investigated in male dogs. Systemic administration of yohimbine caused a biphasic effect on ejaculatory response; the amount of ejaculate produced by the genital stimulation (for 5 min) was dose-dependently increased by low doses (0.01-0.10 mg/kg) of yohimbine, whereas it was decreased by the highest dose (1.00 mg/kg) of yohimbine. The erectile potency was attenuated only, by the highest dose of yohimbine. The most effective dose (0.10 mg/kg) of yohimbine on ejaculation did not affect the duration of penile erection after removing the genital stimulation. In a stereoisomer's testing, the stimulatory effect on ejaculation was also observed by rauwolscine, an alpha-2 adrenoceptor antagonist (0.03 and 0.10 mg/kg), but not by corynanthine, an alpha-1 adrenoceptor antagonist (0.10 and 0.30 mg/kg). These results suggest that yohimbine at low doses specifically facilitate the ejaculatory response through the blockade of the alpha-2 adrenoceptors. This study also indicates that the effects of yohimbine on male genital responses vary with its dosage used.