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Biomedical subjects

K Uchikawa

Publications and source records attributed to K Uchikawa.

At least 19 recordsLinked to original sources

Integration of mutations responsible for the attenuated phenotype of Pepper mild mottle virus strains results in a symptomless cross-protecting strain.

An enhanced attenuated strain of Pepper mild mottle virus (PMMoV) was constructed by incorporating mutations that affect viral attenuation from three reported attenuated strains of PMMoV, which causes serious economic losses in the production of green pepper in Japan. The new strain caused no symptoms on pepper plants and protected them from infection by a wild-type strain. The mutations responsible for viral attenuation were located in the intervening region (IR) of the 126-kDa/183-kDa proteins. The mutations had synergistic effects in terms of the attenuation of symptoms and decreased the accumulation of the viral coat protein in infected pepper plants. In this paper, we propose an efficient method for the improvement of attenuated viruses by reverse genetics in plant viruses.

Base Sequence↗

Apparent size of an object remains uncompressed during presaccadic compression of visual space.

It is well known that compression of visual space occurs near the saccade goal when visual stimuli are briefly flashed at various locations on a visual reference just before a saccade. We investigated how presaccadic compression of visual space affected the apparent size of an object. In the first experiment, subjects were instructed to report the apparent number of multiple bars briefly presented around the time of saccade onset. The reported number of four bars began to decline at the 50 ms mark before a saccade and reached a minimum near the saccade onset. This confirms that the compression of visual space occurs just before saccades. In the second experiment, subjects judged the apparent width of a rectangle (a single element) or four bars (four elements) presented just before saccades. The apparent width of the four-bar stimulus was compressed just before saccades, but that of the rectangle stimulus was not compressed. Experiment 3 shows that the width compression of the four-bar stimulus is consistent with the width change predicted by compression of position. These findings indicate that the shape of a single object is not distorted at the saccade goal during presaccadic compression of visual space. In addition, experiment 4 indicates that the apparent width of a flashed stimulus just before saccades depends on the processing of global shape. This extends the definition of a visual object during presaccadic compression of visual space to not only a solid element but also a constellation of multiple elements. Furthermore, the results from these experiments suggest that presaccadic compression of visual space does not prevent object recognition underlying an attentional mechanism in generating saccadic eye movements.

Adult↗

Free 4-hydroxyproline content in serum of bedridden aged people is elevated due to fracture.

Hydroxyproline (Hyp) content in biological fluids is used as a parameter of collagen catabolism, especially bone resorption. In this paper, we examined the relationship between aging and serum free 4-Hyp and proline (Pro) content using a newly introduced analytical technique which is chemiluminescence determination with electrogenerated tris (2,2'-bipyridine) ruthenium(III). Serum was collected from 50 bedridden aged people and 131 normal subjects living in Yamato-son, Amamioshima, Kagoshima. In the free 4-Hyp content of normal serum, the change in females is more gentle than that males with aging. The free 4-Hyp content of bedridden aged serum is significantly (p<0.05) elevated compared to normal aged serum. Bedridden aged females and males also show levels significantly (p<0.05) elevated in comparison to normal. These changes are due to bone resorption based on bedridden patients' atrophy. In bedridden aged people, free 4-Hyp of those with a fracture was significantly (p<0.05) elevated than in people without a fracture. No relationship between low bone density and high free 4-Hyp in serum was observed. It seems that bone resorption or bedridden atrophy may be enhanced in bedridden aged people by a fracture. In free Pro content in serum, no significant change was observed. These results show that the free 4-Hyp content in serum is useful in the clinical or biological inspection of bone resorption, especially in aged people.

Adult↗

Upper-limit luminance for the surface-color mode appearance.

A series of experiments were carried out to reveal determinants for the mode of color appearance by measuring the upper-limit luminance of a color chip for the surface-color mode. We used a CRT color monitor to present test and surround stimuli in the surface-color mode. The stimuli were composed of a three-by-three array of color chips on a gray background with a white frame. The observer increased the luminance of a center test color until it just ceased to appear in the surface-color mode. Our results show that this upper-limit luminance was different among test colors, but their brightnesses, calculated from the luminance and brightness/ luminance values, were almost the same and were slightly below the brightness of the white frame. The existence of the surrounding color chips affected the results, but their sizes and spatial arrangements did not. When all of the luminances of the surrounds changed equally, the upper-limit luminances of the test colors for the surface-color mode appearance changed by the same ratio. This result indicates that the brightness of a target was a determinant for selecting the mode of color appearance and that the brightest surround stimulus acted as a cue for determining the judgment.

Journal Article↗

Increment-threshold spectral sensitivity during saccadic eye movements in uniform visual field.

We measured increment-threshold spectral sensitivity functions during saccades, immediately after saccades and during fixation. A uniform white background field which covered observer's whole visual field prevented any retinal masking effects. Visual sensitivity was degraded during saccades or after saccades. The reduction in sensitivity depended upon the wavelength of the test stimulus. The spectral sensitivity function during fixation produced a broad smooth curve in the middle and long wavelength region, while saccades caused a prominent dip around 570 nm. This finding indicates that saccadic suppression, which cannot be attributed to retinal masking, has more effect on the achromatic channel than the chromatic channel. The role of central and retinal processes dealing with the perceptual clearness and stableness across saccades will be discussed.

Adult↗

Crystal structure of prolyl aminopeptidase from Serratia marcescens.

Prolyl aminopeptidase from Serratia marcescens specifically catalyzes the removal of N-terminal proline residues from peptides. We have solved its three-dimensional structure at 2.3 A resolution by the multiple isomorphous replacement method. The enzyme consists of two contiguous domains. The larger domain shows the general topology of the alpha/beta hydrolase fold, with a central eight-stranded beta-sheet and six helices. The smaller domain consists of six helices. The catalytic triad (Ser113, His296, and Asp268) is located near the large cavity at the interface between the two domains. Cys271, which is sensitive to SH reagents, is located near the catalytic residues, in spite of the fact that the enzyme is a serine peptidase. The specific residues which make up the hydrophobic pocket line the smaller domain, and the specificity of the exo-type enzyme originates from this smaller domain, which blocks the N-terminal of P1 proline.

Aminopeptidases↗

Apparent depth with retinal image motion of expansion and contraction yoked to head movement.

The two main questions addressed in this study were (a) what effect does yoking the relative expansion and contraction (EC) of retinal images to forward and backward head movements have on the resultant magnitude and stability of perceived depth, and (b) how does this relative EC image motion interact with the depth cues of motion parallax? Relative EC image motion was produced by moving a small CCD camera toward and away from the stimulus, two random-dot surfaces separated in depth, in synchrony with the observers' forward and backward head movements. Observers viewed the stimuli monocularly, on a helmet-mounted display, while moving their heads at various velocities, including zero velocity. The results showed that (a) the magnitude of perceived depth was smaller with smaller head velocities (< 10 cm s-1), including the zero-head-velocity condition, than with a larger velocity (10 cm s-1), and (b) perceived depth, when motion parallax and the EC image motion cues were simultaneously presented, is equal to the greater of the two possible perceived depths produced from either of these two cues alone. The results suggested the role of nonvisual information of self-motion on perceiving depth.

Adult↗

Roles of the Ser146, Tyr159, and Lys163 residues in the catalytic action of 7alpha-hydroxysteroid dehydrogenase from Escherichia coli.

The Escherichia coli 7alpha-hydroxysteroid dehydrogenase (7alpha-HSDH; EC 1.1.1.159) has been the subject of our studies, including the cloning of its gene, and determination of the crystal structures of its binary and ternary complexes [J. Bacteriol. 173, 2173-2179 (1991); Biochemistry 35, 7715-7730 (1996)]. Through these studies, the Ser146, Tyr159, and Lys163 residues were found to be involved in its catalytic action. In order to clarify the roles of these residues, we constructed six single mutants of 7alpha-HSDH, Tyr159-Phe (Y159F), Tyr159-His (Y159H), Lys163-Arg (K163R), Lys163-Ile (K163I), Ser146-Ala (S146A), and Ser146-His (S146H), by site-directed mutagenesis. These mutants were overexpressed in E. coli WSD, which is a 7alpha-HSDH null strain, and the expressed enzymes were purified to homogeneity. The kinetic constants of the mutant enzymes were determined, and the structures of the Y159F, Y159H, and K163R mutants were analyzed by X-ray crystallography. The Y159F mutant showed no activity, while the Y159H mutant exhibited 13.3% of the wild-type enzyme activity. No remarkable conformational change between the Y159F (or Y159H) and wild-type proteins was detected on X-ray crystallography. On the other hand, the K163I mutant showed just 5.3% of the native enzyme activity, with a 8. 5-fold higher Kd. However, the K163R mutant retained 64% activity, and no remarkable conformational change was detected on X-ray crystallography. In the cases of the S146A and S146H mutants, the activities fairly decreased, with 20.3 and 35.6% of kcat of the wild-type, respectively. The data presented in this paper confirm that Tyr159 acts as a basic catalyst, that Lys163 binds to NAD(H) and lowers the pKa value of Tyr159, and that Ser146 stabilizes the substrate, reaction intermediate and product in catalysis.

Binding Sites↗

Adaptive shift of visual sensitivity balance under ambient illuminant change.

We examined the relationship between the ambient illuminant chromaticity and changes in the sensitivity balance of the visual system, using illuminants of various chromaticities. The sensitivity of observers was measured in a room with a variable-chromaticity illuminant. The observer's state of chromatic adaptation was measured with unique-white settings. Our results showed that the change in visual sensitivity has a nonlinear correlation with the change in illuminant chromaticity; chromatic adaptation was nearly complete for desaturated illuminants, but the degree of chromatic adaptation became worse as the illuminant became more saturated. We defined a new index, relative cone weights, which represents this relationship well. To measure the role of chromatic induction from the immediate-surround area of the matching stimulus, we performed additional experiments by presenting the test inside a colored or black immediate surround. The results showed that the unique-white settings were not disturbed by the change in immediate-surround color. Our results imply that the room illuminant chromaticity was the primary factor in changing the observer's state of chromatic adaptation.

Adaptation, Physiological↗

Perceived angular and linear size: the role of binocular disparity and visual surround.

An experiment was conducted to investigate the effects of perspective cue and binocular disparity on perceived angular and linear size. Following the 'angular' and 'linear' instructions, subjects matched the size of two squares, for which the binocular disparity between the centers of the squares and the configuration of the stimulus surrounding the squares were manipulated. Results showed that angular-size matches depended on the retinal-image size and the binocular disparity, and not on the visual-surround stimulus. Linear-size matches, on the other hand, depended on the visual-surround stimulus as well as on the image size and the binocular disparity. The visual-surround stimulus also affects the perceived depth between the test squares. These findings indicate that perceived angular and linear size depend on different processes that use different cues, and suggest that there is a causal relationship between perceived depth and perceived linear size.

Adult↗

Distribution of Borrelia species associated with Lyme disease in the subalpine forests of Nagano prefecture, Japan.

We surveyed the natural distribution of Borrelia species associated with Lyme disease in the subalpine forests of Nagano prefecture, Japan, during 1993-94. Tick-derived isolates (n = 112) from Ixodes persulcatus and rodent-derived isolates (n = 55) from Apodemus argenteus, Apodemus speciosus, Eothenomys andersoni, Eothenomys smithii, and Microtus montebelli were classified by rRNA gene restriction fragment length polymorphism analysis (RFLP ribotyping). Ribotype group IV (an intraspecific variant of Borrelia garinii) was predominant among the tick isolates. It was also isolated repeatedly from the rodents. Ribotype group III (Borrelia afzelii) was detected in low frequencies among the tick and rodent isolates. The data suggest that humans are likely to be exposed to the group IV when they are bitten by I. persulcatus ticks.

Animals↗

Limitations of surface-color and apparent-color constancy.

Color-constancy mechanisms have been studied and discussed in a number of investigations. However, there has been little attempt to reveal how color constancy deteriorates as the conditions for it become less than optimal. We carried out a series of asymmetric color-matching experiments, using two criteria: surface-color match and apparent-color match. With brief adaptation the degree of color constancy increased as chromatic cues were added in the surround. In the condition of black surround, the test stimuli appeared self-luminous, and chromaticities of the chosen matching stimuli were the same as the physical chromaticities of the test stimulus, indicating a total deficiency of color constancy. With 15 min of preadaptation to the illuminant, the surface-color matches showed almost perfect color constancy under illuminant change. In both adaptation conditions, the chromatic-shift of matches from what would be expected for perfect color constancy increased gradually between 1,700- and 30,000-K illuminant, as chromaticity of the illuminant departed from 6,500-K illuminant. Under 1,000-K illuminant the surface-color appearance became totally achromatic, and color constancy was completely lost. Our results show that, even with brief adaptation to the illuminant, the contribution of the surrounding stimulus is large enough to achieve a fair degree of color constancy, but complete adaptation to the illuminant helps to achieve almost perfect color constancy.

Adaptation, Physiological↗

Saccadic suppression of achromatic and chromatic responses measured by increment-threshold spectral sensitivity.

We measured spectral-sensitivity functions during saccadic eye movement by the increment-threshold method to test whether saccades selectively suppressed achromatic or chromatic responses. A circular monochromatic test stimulus of 12-deg diameter was presented for 10 ms on a 62 deg x 43 deg white background, and observations were made under three conditions: during fixation, during 6-deg saccades, and immediately after saccades. In two additional conditions the test stimulus was made to move during fixation and during 6-deg saccades at the same speed and in the same direction as the saccades. The during-fixation spectral-sensitivity function was found to resemble the relative luminous efficiency V (lambda) function in shape except for the case of short wavelengths, whereas the during-saccade spectral-sensitivity function showed lower sensitivity for all wavelengths and had three prominent peaks at approximately 440, 530, and 600 nm. These characteristics did not depend on whether the stimulus was stationary or moving. These results indicated that saccadic suppression was greater for achromatic than for chromatic response. A possible suppression mechanism was discussed involving the magno and parvo pathways.

Adult↗

Trombiculid fauna and seasonal abundance of Leptotrombidium scutellare (Acari: Trombiculidae) in an endemic area of scrub typhus (Tsutsugamushi disease) in Yamakita Town, Kanagawa Prefecture, Japan.

Using the Tullgren funnel method, we surveyed for trombiculid larvae around dwellings and in mandarin orange groves on the slopes of Sengenyama and Maruyama, Yamakita Town, Kanagawa Prefecture, Japan, during the period between 8 October 1992 and 8 January 1993. In total, 6,690 trombiculid larvae, consisting of 6,669 Leptotrombidium scutellare (Nagayo, Mitamura, Tamiya & Tenjin), 13 L. fuji Kuwata, Berge & Philip, two L. himizu (Sasa, Kumada, Hayashi, Enomoto, Fukuzumi & Obata), two Neotrombicula japonica (Tanaka, Kaiwa, Teramura & Kagaya), one Gahrliepia saduski Womersley, and three Walchia ogatai Sasa & Teramura, were taken from 240 soil samples (40 samples, six times at intervals of 2-4 wk). From the above species composition, L. scutellare is suspected to be the sole vector of scrub typhus in the survey area, where the disease is known to be caused by two serotypic strains of Rickettsia tsutsugamushi Ogata, Kawasaki and Kuroki. L. scutellare larvae rapidly increased in number in late October to form an acuminate peak of abundance in early November, followed by a gradual wane.

Agriculture↗

Infectivity and early antibody response to Borrelia burgdorferi sensu lato isolated in Japan in outbred mice.

Borrelia burgdorferi sensu lato isolated from Ixodes ovatus (B. japonica), I. persulcatus and patients with erythema migrans (EM) in Japan were determined on infectivity and arthritis induction-activity in outbred mice. Infectivity of B. japonica was weak and did not induce the development of footpad swelling by subcutaneous (s.c.) inoculation into the footpad. Challenged strain, NO129-M of B. japonica, to ddY mice were reinoculated to the mice at various cell numbers (1 x 10-1 x 10(6) cells/mouse). The strain isolated from the mouse did not reinfect ddY mice and did not induce the production of specific antibody to the homologous strain. On the other hand, strains from I. persulcatus and patients with EM in Japan infected the mice and induced a serious inflammatory response in Borrelia-inoculated footpad as well as strains belonging to the three genospecies, B. burgdorferi sensu stricto, B. garinii, and B. afzelii, related to Lyme disease, from North America and Europe. The mice were infected with 10 cells of strain HP1 isolated from I. persulcatus in Hokkaido and of strain 297 isolated from a patient in the U.S.A. by subcutaneous inoculation into the hind footpad, or by intradermal inoculation into the back. Antigens of ca. 20, 23-24 (Osp C), 29, 39, 41 (flagellin) and 45 kDa reacted with the pooled sera from mice inoculated with strains HP1 and 297, but Osp A and Osp B did not.

Animals↗

Correlation of octanol-water partition coefficients with capacity factors measured by micellar electrokinetic chromatography.

Micellar electrokinetic chromatography (MEKC) was used to measure hydrophobicity of 18 aromatic compounds. The capacity factors, which are proportional to the micelle-water distribution coefficients, were closely correlated with the octanol-water partition coefficients. The addition of Brij 35 to the charged micelle was effective in improving the correlation because the surface charges of the micelle were shielded.

Chromatography↗