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H Oyamada

Publications and source records attributed to H Oyamada.

At least 37 records · Page 2Linked to original sources

Characterization of beta-1,2-mannosyltransferase in Candida guilliermondii and its utilization in the synthesis of novel oligosaccharides.

A particulate insoluble enzyme fraction containing mannosyltransferases from Candida guilliermondii IFO 10279 strain cells was obtained as the residue after extracting a 105,000 x g pellet of cell homogenate with 1% Triton X-100. Incubation of this fraction with a mannopentaose, Manalpha1-->3(Manalpha1-->6)Manalpha1-->2Manalpha1+ ++-->2Man, in the presence of GDP-mannose and Mn2+ ion at pH 6.0 gave a third type of beta-1,2 linkage-containing mannohexaose, Manbeta1-->2Manalpha1-->3(Manalpha1-->6)Manalpha1++ +-->2Manalpha1-->2Man , the structure of which was identified by means of a sequential NMR assignment. The results of a substrate specificity study indicated that the beta-1,2-mannosyltransferase requires a mannobiosyl unit, Manalpha1--> 3Manalpha1-->, at the nonreducing terminal site. We synthesized novel oligosaccharides using substrates possessing a nonreducing terminal alpha-1,3-linked mannose unit prepared from various yeast mannans. Further incubation of the enzymatically synthesized oligosaccharide with the enzyme fraction gave the following structure, Manbeta1-->2Manbeta1-->2Manalpha1-->3(Manalpha1- ->6)Manalpha1--> 2Manalpha1-->2Man, which has been found to correspond to antigenic factor 9. Incubation of Candida albicans serotype B mannan with the enzyme fraction gave significantly transformed mannan, which contains the third type of beta-1,2-linked mannose units.

Antigens, Fungal↗

Structure of a cell wall mannan from the pathogenic yeast, Candida catenulata: assignment of 1H nuclear magnetic resonance chemical shifts of the inner alpha-1,6-linked mannose residues substituted by a side chain.

We performed an enzyme-linked immunosorbent assay of the cell wall mannan purified from the pathogenic yeast, Candida catenulata, using antisera to factors of the genus Candida. The results suggest that mannan possesses a linear backbone consisting of alpha-1, 6-linked mannose residues and side chains possessing nonreducing terminal alpha-1,2- and alpha-1,3-linked mannose residues. The chemical structure of the mannan was analyzed by two-dimensional homonuclear Hartmann-Hahn and two-dimensional nuclear Overhauser enhancement and exchange spectroscopy. The sequential assignments of the cross-peaks caused by J-coupling and the nuclear Overhauser effect from these terminal mannose residues demonstrate that the H1 signal of an inner alpha-1,6-linked mannose residue substituted by an alpha-oligomannosyl side chain or a single mannose through the C-2 position in an alpha-anomer configuration undergoes a significant downfield shift (delta delta = 0.16 or 0.19 ppm, respectively) compared with that of unsubstituted residues. We therefore propose the exact overall structure of the antigenic mannan obtained from C. catenulata. The assignment data in the present study are useful for the determination of the exact overall structure of various yeast mannans using the two-dimensional nuclear magnetic resonance analysis without the need for harsh procedures.

Antigens, Fungal↗

Expression of mRNA for RANTES in human eosinophils.

RANTES has been considered to play an important role in various immune and allergic disorders since RANTES is a potent chemoattractant for various important inflammatory cells such as eosinophils. Eosinophils, on the other hand, are considered to be the major inflammatory cells in bronchial asthma since eosinophil-specific granule proteins can damage bronchial mucosal cells. The recent demonstration that eosinophils themselves could synthesize some cytokines such as IL-5 indicates the role of these cytokines not only in paracrine but also in autocrine regulation of eosinophil production, differentiation and activation. Therefore, in this study, we examined mRNA for RANTES and release of RANTES by human eosinophils. The present findings revealed that eosinophils obtained from asthmatic patients could express and release RANTES. Taken together, these findings suggest that eosinophils play a crucial role in the pathogenesis, particularly in eosinophil and T lymphocyte recruitment into the inflamed sites in asthma through RANTES production.

Asthma↗

Identification of the antigenic determinants of factors 8, 9, and 34 of genus Candida.

We investigated the antigenic determinants of factors 8, 9, and 34 of the genus Candida among pathogenic yeasts by enzyme-linked immunosorbent assay (ELISA) using mannans of Saccharomyces cerevisiae wild type and mutant types, mnn 1-mnn 4 and mnn 2. Results of ELISA including antisera against the antigenic factors of genus Candida (Candida Check, latron; FAbs) indicated that these three types of mannan distinctly react with FAbs 34, 8 and 9, respectively. To identify the recognition sites of these FAbs, we compared the ability of various oligosaccharides to inhibit the binding of the mannans to FAbs. The results indicated that FAb 34 preferentially recognizes linear side chains containing a non-reducing terminal alpha-1,3-linked mannose residue, Man(alpha)1 --> 3Man(alpha)1 --> (2Man(alpha)1 --> )n(2Man) (n > or = 0), and that one of the recognition sites of FAb 9 is linear alpha-1,6-linked oligomannosyl series, Man(alpha)1 --> (6Man(alpha)1 --> )n(6Man) (n > or = 2). On the other hand, the recognition site of FAb 8 apparently consisted of two alpha-1,2-linked oligomannosyl side chains and an alpha-1,6-linked mannose residue that originated from the mannan backbone, Man(alpha)1 --> 2Man(alpha)1 --> 2(Man(alpha)1 -->2Man(alpha)1 --> 6)Man.

Antigens, Fungal↗

Characterization of alpha-1,6-mannosyltransferase responsible for the synthesis of branched side chains in Candida albicans mannan.

A particulate insoluble fraction from Candida albicans NIH B-792 (serotype B) strain cells was obtained as the residue after extracting a 105000 x g pellet of cell homogenate with 1% Triton X-100. Incubation of this fraction with a mannopentaose, Man alpha 1-->3Man alpha 1-->2Man alpha 1-->Man alpha 1-->2Man, in the presence of GDP-mannose and Mn2+ at pH 6.0 gave a branched mannohexaose, [sequence: see text] 6 the structure of which was identified by means of sequential off assignment. However, the enzyme fraction obtained from Candida parapsilosis gave Man alpha 1-->2Man alpha 1-->3Man alpha 1-->2Man alpha 1-->2 Man alpha 1-->2Man under the same conditions. These results demonstrate the finding that the structural difference in the mannans of these two species is due to the presence of alpha-1.6-linked branching mannose units in the C. albicans mannan [Shibata, N., Ikuta, K., Imai, T., Satoh, Y., Satoh, R., Suzuki, A., Kojima, C., Kobayashi, H., Hisamichi, K. & Suzuki, S. (1995) J. Biol. Chem. 270, 1113-1122]. The substrate-specificity study of the enzyme indicated that the structural requirement of the alpha-1,6-mannosyltransferase is Man alpha 1-->3Man alpha 1-->. The alpha-1,6-mannosyltransferase also transferred the alpha-1,6-linked branching mannose unit to the mannan of Saccharomyces cerevisiae. The transformation of the mannan was detected by the appearance of antigenic factor 4 using an enzyme-linked immunosorbent assay and two-dimensional homonuclear Hartmann-Hahn spectroscopy.

Candida↗

Changes in oculomotor functions before and after loading of a 3-D visually-guided task by using a head-mounted display.

Changes in visual and oculomotor functions were tested in eight young volunteer subjects after performing a 3-D visually-guided task for 25 min. The visual stimuli were given by using a head-mounted display. No significant change was detected in the AC/A ratio and the stereo acuity. Changes were found in refraction and ocular convergence. Slight but significant hyperopic changes were detected in refraction after the task. The mean amplitude of convergence eye movement elicited by test stimuli after the task was significantly changed in the pooled data. They were significantly reduced when the subjects were tested by the disparity step of 0.7 degrees, but significantly increased when tested by the step of 6 degrees. The mean amplitudes were not significantly changed after the task when the subjects were tested by the intermediate disparity steps of 1.5 and 3 degrees. In data obtained for each subject, the amplitude of ocular convergence evoked by test stimuli after the task was reduced significantly in most subjects. In the majority of subjects, however, the results were not consistently significant when they were tested by step stimuli with different disparities. In only a few subjects, the changes were consistently significant except when the largest disparity was tested. On the other hand, the mean peak velocities of ocular convergence after the task were not significantly changed in the pooled data. In individual subjects, the changes of velocities of ocular convergence evoked by test stimuli after the task were more variable: they increased in some subjects but decreased in other subjects. Consequently, in only one subject, both amplitudes and peak velocities of ocular convergence tested by the disparity steps of 0.7, 1.5 and 3 degrees were consistently reduced after the task. The changes in refraction or ocular convergence found in this study were relatively small, and were not in the pathological range. The significance of these data are discussed. The results of the present study provide the basis for the more detailed analyses of the human factor in the head-mounted display.

Adult↗

Enhancement of the antitumor effect of gamma-ray irradiation in combination with camptothecin against human colorectal adenocarcinoma.

The effect of gamma-ray irradiation and camptothecin (CAM) on human colorectal adenocarcinoma (LS 174T) growth was examined. On inhibition of DNA synthesis of LS 174T cells, almost all combined treatments presented a synergistic effect, excepting that the additive effects were shown at the lowest or the highest doses of CAM prior to the gamma-ray (137Cs) irradiation. The in vivo antitumor activity of 131I-NCC-CO-411 monoclonal antibody against carcinoembryonic antigen in combination with CAM was assessed by a tumor growth rate in LS 174T tumor-bearing nude mice. The tumor growth of the combined group was supra-additively suppressed in comparison with that of the individual group. The results suggest that the efficacy of radioimmunotherapy may be enhanced by combination treatment with CAM. A clinical protocol involving a combination treatment with CAM is underway.

Adenocarcinoma↗

Accumulation of NCC-CO-411 monoclonal antibody in human colorectal adenocarcinoma through binding of the released carcinoembryonic antigen in nude mice.

NCC-CO-411, an anti-carcinoembryonic antigen (CEA) monoclonal antibody (MoAb) showed a dose-dependent reactivity with standard CEA, but did not bind to the CEA anchoring with cell membranes. When LS 174T cells, one of the CEA-producing cells, were treated with phosphatidylinositol-specific phospholipase C, NCC-CO-411 MoAb recognized the released CEA containing glycosylphosphatidylinositol as well as the standard CEA. The binding activity of NCC-CO-411 MoAb with CEA was also demonstrated by the immunostaining of the LS 174T colorectal tumor xenograft tissues. The biodistribution study showed that NCC-CO-411 MoAb was mainly taken up by tumor and kidney, while the anti-hCEA MoAb was largely concentrated in liver and spleen rather than in the tumor. These results suggest that NCC-CO-411 MoAb recognizes the released CEA and gives an excellent tumor image in LS 174T tumor-bearing mice.

Adenocarcinoma↗

[Effects on visual functions following several hours' usage of a head mounted display].

We investigated the effects of viewing video movies with a head-mounted display (HMD) for 4 to 6 hours on visual functions such as refraction, visual acuity, and accommodation-vergence system. Two or three video movies were watched without any breaks by 13 normal volunteers (age: 22 approximately 40). Measurements were made of (1) objective and subjective refraction, (2) corrected visual acuity, (3) tonic level and step response of accommodation with a computer-assisted infrared optometer, and (4) near and far phorias and AC/A ratio. Significant transient myopia was found following 4 hours' viewing, but not following 6 hours' viewing. Scrutinizing individual data, myopia was consistently found in some subjects, and hyperopia in others. We presumed that many subjects might have been influenced by initial instrumental myopia when they adjusted the focus by using the mechanism built in the HMD. No significant change was observed in any other examination. However, there was a tendency for the AC/A ratio to change after a short time, and then to recover to its original value. Based on the results in this study, it appears that some changes in accommodation and vergence systems are caused by viewing video movies with the HMD. Although the amount of changes was within normal physiological variation in this study, the possibility still remains that usage for a longer time may lead to other changes in visual function. Care is also necessary when using the HMD in subjects with subclinical problems.

Accommodation, Ocular↗

[Diagnostic accuracy of 201T1 SPECT.MRI in brain diseases and inter-reader variance of diagnostic performance].

Fifteen-four studies of 201T1 brain tumor SPECT were independently interpreted by 9 nuclear medicine physicians with and without reference magnetic resonance images in 2 separate sessions to define an effect of referring images, and inter-observer variations. The physicians were requested to detect foci of abnormal deposits, and to discriminate whether they were malignant or not according to 5-grade scaling of subjective diagnostic confidence. Receiver-operating characteristics (ROC) analysis was performed. Mean sensitivity for presence of lesions (SEP), and sensitivity and specificity for malignancy of 201T1 SPECT were 84, and 53 and 55%, which were changed to 94, and 74 and 55% after referring to the MR images. The SFP was significantly improved (p < 0.05), but sensitivity and specificity for malignancy, assessed by areas under the ROC curves, were not significantly improved. Inter-observer variation of the SFP was significantly reduced with addition of the referring MR images. Cerebral lobar localization of lesions by SPECT showed great inter-observer variations (true localization ranged from 30 to 68.4%). It is concluded that 201T1 brain tumor SPECT has moderate sensitivity and specificity for malignancy, which is not improved by addition of anatomical reference images, that additional MR images reduce inter-observer variation of confidence on lesion presence, and that SPECT localization of lesions has great inter-observer variations.

Adolescent↗

Postnatal development of the substance P-, neuropeptide Y- and serotonin-containing fibers in the rat suprachiasmatic nucleus in relation to development of the retino-hypothalamic projection.

The suprachiasmatic nucleus (SCN) in the hypothalamus controls many of the circadian rhythms in mammalian species. In the present study, we investigated the development of substance P (SP)-, neuropeptide Y (NPY)- and serotonin (5-hydroxytryptamine, 5-HT)-immunoreactive fibers in the rat SCN and the development of the retino-hypothalamic tract using cholera toxin beta subunits (CTB), in order to understand which parts of the SCN participate in diurnal rhythm regulation and entrainment. In newborn rats, SP-, NPY- and 5-HT-immunoreactive fibers were scarcely detected in the SCN. The number of SP-immunoreactive fibers gradually increased between postnatal days (P) 15 and 30. At P30, the distribution pattern of SP-immunoreactive fibers in the SCN was similar to that in the adult rat. The number of NPY- and 5-HT-immunoreactive fibers increased greatly between P10 and P15, and the increase in NPY- and 5-HT-immunoreactive fibers continued until P20. CTB was injected into the unilateral eyeball of the rat at various postnatal stages. In neonates, several labeled retinal fibers already existed in the ventral part and ventro-lateral edge of the SCN. The number and density of labeled retinal fibers in the SCN gradually increased between P10 and P20. Between P20 and P30, a decrease in the labeling was observed in the dorsolateral part of the SCN. The adult pattern of labeled retinal fibers was achieved between P20 and P30. The development of SP-immunoreactive fibers was delayed about 10 days relative to that of NPY-, 5-HT-immunoreactive fibers and retinal fibers.

Animals↗

Antitumor effect of ischemia-reperfusion injury induced by transient embolization.

The effect of ischemia-reperfusion, induced by the transient embolic agent degradable starch microspheres (DSMs) on tumor tissue was investigated from the standpoint of active oxygen species. Rabbits with VX2 carcinoma received regional infusion of DSMs by transcatheter angiography, and it was confirmed that DSMs occluded tumor vessels. Blood flow in the tumors decreased rapidly immediately after the DSM treatment and returned to the original level within 40 min. The size of tumors did not change after a single infusion of DMS, while five repeated DSM treatments led to a significant reduction in tumor size. This reduction in tumor size was prevented by the treatment of rabbits with superoxide dismutase and catalase, indicating that the generation of active oxygen species in the tumor was involved in the mechanism of action of DSMs. Thiobarbituric acid-reactive substances also increased in the tumors after DSM infusion, and this increase was also inhibited by treatment with superoxide dismutase and catalase. In conclusion, the antitumor effect of the transient embolic agent DSM is secondary to the phenomenon of ischemia-reperfusion injury. In addition, active oxygen species and lipid peroxidation are possible causes of ischemia-reperfusion injury.

Animals↗

Primary structure and distribution of ryanodine-binding protein isoforms of the bullfrog skeletal muscle.

We have cloned two groups of cDNAs which encode isoforms of ryanodine-binding protein/Ca2+ release channel of the bullfrog skeletal muscle sarcoplasmic reticulum. One of the cDNA groups encodes the protein of 5,037 (or 5,031 with a deletion) amino acids with a molecular weight of 571,262 (or 570,607), which is identified as the alpha-isoform of the ryanodine-binding protein based on the amino acid sequence of three tryptic fragments of the purified protein. The other group of cDNAs encodes the protein of 4,868 amino acids with molecular weight of 553,029, which contains the sequences of three proteolytic fragments derived from the beta-isoform protein. About 70% of the amino acid sequence identity is present between alpha- and beta-isoforms of the bullfrog. The primary structure of the alpha-isoform is highly (80%) homologous to the ryanodine-binding protein cloned from rabbit skeletal muscle (type 1). The beta-isoform, on the other hand, is more than 85% identical with that from the rabbit brain (type 3), while it has only 67% overall identity with type 1. Analyses of RNA from various tissues of the bullfrog demonstrate that the beta-isoform is widely expressed, while the alpha-isoform is expressed mainly in skeletal muscle. A phylogenetic analysis of the ryanodine-binding protein/Ca2+ release channel family suggests that the various types of Ca2+ release channels have evolved from an ancestor gene. Possible differential roles of alpha- and beta-isoforms of ryanodine-binding protein in Ca2+ release mechanisms including skeletal muscle excitation-contraction coupling were discussed.

Animals↗

Reliability of data obtained by radionuclide angiocardiography in follow-up studies with special reference to intra- and interobserver variations.

The reproducibility of the data obtained by radionuclide angiocardiography was studied in terms of the intra- and intervariations of three technicians. Three parameters were chosen: the left ventricular ejection fraction (LVEF), the 1/3 filling fraction (1/3FF), and the ratio of the time from end systole to the point of peak filling rate against the entire diastolic period (TRPFR). First, each technician studied an independent consecutive series of 40 patients for the initial analysis. The original data of each patient, composed of 26 frames for one beat, were stored on an optical disc for the intra- and intervariation studies. In this study, analysis of the volume curve was performed by using both 3rd order and 4th order Fourier series. The region of interest (ROI) for the left ventricle, which was set for the initial analysis, was recorded by Polaroid photography to be used as a reference for the later analyses for the variation studies. The least variation was noted in the LVEF not only for the intravariation study but also for the intervariation study, no matter which order of the Fourier series was used. However, the 3rd order Fourier series seemed to give better curve fitting to avoid fluctuation of the diastolic parameters. The results indicate that we can expect more steady and reliable information from LVEF than from the diastolic parameters, even when various technicians perform the follow-up study of a particular patient.

Angiocardiography↗

Structure of cell wall mannan of Candida kefyr IFO 0586.

We conducted a structural analysis of the antigenic cell wall mannoprotein (mannan) isolated from Candida kefyr (formerly Candida pseudotropicalis) IFO 0586. The result of two-dimensional homonuclear Hartmann-Hahn analysis of this mannan indicates that the molecule is constructed from alpha-1,2- and alpha-1,6-linked mannopyranose residues. Upon alkali treatment (beta-elimination reaction), this mannan released two alpha-1,2-linked mannooligosaccharides, biose and triose. The structure of the alkali-stable mannan (outer chain) moiety was investigated by acetolysis. The structures of the resultant oligosaccharides, biose and triose, from the outer chain moiety were found to be the same as those of the alkali-released ones. Further, the treatment of the parent mannan with an Arthrobacter GJM-1 exo-alpha-mannosidase gave a linear mannan consisting solely of alpha-1,6-linked mannopyranose residues. These results indicate that the mannan forms the long backbone of the alpha-1,6 linkage, with a large number of short alpha-1,2-linked oligomannosyl side chains forming a comblike structure. Moreover, we investigated the serological properties of this mannan by performing an inhibition assay of a slide agglutination reaction with mannooligosaccharides and polyclonal factor sera (Candida Check; Iatron). The result indicates that the factor 1 serum preferentially recognizes the alpha-1,2-linked oligomannosyl side chains in this mannan. On the other hand, the fact that the mannan does not contain an antigenic determinant(s) corresponding to factor 8 suggests that the epitope(s) of this factor resides in other molecules on the cell surface of this strain.

Candida↗