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Biomedical subjects

M Matsuoka

Publications and source records attributed to M Matsuoka.

At least 37 records · Page 2Linked to original sources

KNOX homeodomain protein directly suppresses the expression of a gibberellin biosynthetic gene in the tobacco shoot apical meristem.

To identify genes targeted by the tobacco KNOX homeodomain protein, Nicotiana tabacum homeobox 15 (NTH15), we have generated an inducible system using the human glucocorticoid receptor. In this system, steroid treatment strictly induced NTH15 function and immediately suppressed the expression of a gibberellin (GA) biosynthetic gene encoding GA 20-oxidase (Ntc12) and also resulted in a decrease in bioactive GA levels. The repression of Ntc12 was observed even when indirect effects were blocked by cycloheximide. NTH15 mRNA was present in corpus cells of the shoot apical meristem (SAM), whereas Ntc12 mRNA was observed in leaf primordia and rib meristem but not in the corpus. Recombinant NTH15 protein strongly bound to a 5-bp dyadsymmetric sequence, GTGAC, in the first intron of Ntc12 in vitro. Mutation of this sequence in the Ntc12 gene abolished the NTH15-dependent suppression of Ntc12 in the corpus of the SAM. Our results indicate that NTH15 directly represses Ntc12 expression in the corpus of the wild-type SAM to maintain the indeterminate state of corpus cells. The suppression of NTH15 within cells at the flanks of the SAM permits GA biosynthesis, which promotes organized cell proliferation and consequently induces the determination of cell fate.

Cell Division↗

Preparation of Ti-Si binary oxide thin film photocatalysts by the application of an ionized cluster beam method.

Ti-Si binary oxide thin films with various Ti contents were prepared in a dry process by using an Ionized Cluster Beam (ICB) deposition method with multi ion sources. From the results of UV-VIS measurements, the transmittance of these binary oxide thin films is very high compared to pure TiO2 thin films, indicating that the Ti-oxide exists in a highly dispersed state in the SiO2 matrices. The EXAFS spectra of these thin films with lower Ti content exhibited only a sharp preedge peak attributed to the highly dispersed Ti-oxide species which differs from the peaks of anatase TiO2 thin films. UV light irradiation of these Ti-Si binary oxide thin films in the presence of NO were found to lead to the evolution of N2 and O2 with a good linearity against the irradiation time. The lower the Ti content, the higher the reactivity and the selectivity for the formation of N2 were. These results clearly indicate that the highly dispersed Ti-oxide species plays an important role in the formation of N2 and O2 for the photocatalytic decomposition of NO under UV light irradiation.

Journal Article↗

Characterization and photocatalytic reduction of CO2 with H2O on Ti/FSM-16 synthesized by various preparation methods.

Ti/FSM-16 photocatalysts were prepared by various methods and their reactivity for the photocatalytic reduction of CO2 with H2O at 323 K was investigated. UV irradiation of Ti/FSM-16 in the presence of CO2 and H2O led to the formation of CH4 and CH3OH. The photocatalytic reactivity and selectivity differed remarkably by variations in the preparation method. Analyses of the UV-Vis absorption and XAFS spectra showed that the dispersion and local structure of the Ti-oxide species greatly depended on the preparation method. The formation of CH3OH was found to be related to the local structure of the Ti-oxide species while the yield of the photoluminescence was attributed to the charge transfer excited state of the highly dispersed tetrahedrally coordinated Ti-oxide species. Methanol formation was found to be more efficient on Ti/FSM-16 having such highly dispersed tetrahedrally coordinated Ti-oxide species.

Journal Article↗

XAFS studies on the local structures of Ti-HMS mesoporous molecular sieves and their photocatalytic properties.

Various Ti-containing HMS mesoporous molecular sieves having high surface areas and pore volumes have been prepared at ambient temperature. The Ti-HMS samples exhibited a well-defined XRD pattern of typical hexagonal lattice. UV-vis and XAFS spectral for the calcined forms of Ti-HMS indicated that the Ti ions are tetrahedrally coordinated in the framework of HMS for the samples of Ti-HMS(1) and Ti-HMS(2), while at least a fraction of Ti-oxides in Ti-HMS(10) is in octahedral coordination. UV irradiation of the Ti-HMS catalysts in the presence of NO was found to lead to the decomposition of NO to form N2, O2, and N2O at 273 K with different yields and product distributions. The photocatalytic performance of Ti-oxides appears to be completely modified by their incorporation structure and reaction environment. High efficiency and selectivity for the formation of N2 and O2 in the photocatalytic decomposition of NO was achieved with the Ti-HMS(1) having highly dispersed isolated tetrahedral titanium oxide species, while the reactivity decreased with an increase in the Ti content.

Journal Article↗

Local structure of Pb(II) ion catalysts anchored within zeolite cavities and their photocatalytic reactivity for the elimination of N2O.

The Pb2+/ZSM-5 catalyst was prepared by an ion-exchange method and its photocatalytic activity for the decomposition of N2O under UV irradiation was investigated. In-situ UV-Vis absorption spectroscopy and XAFS (XANES and FT-EXAFS) investigations revealed that the Pb2+ ions exist in a highly dispersed state within the pores of the zeolites. UV irradiation of the catalysts in the presence of N2O led to the photocatalytic decomposition of N2O into N2 at temperatures as low as 298K. The effective wavelength of the irradiated UV light indicated that the excited state of the Pb2+ ions included within the zeolite cavities plays a significant role in the photocatalytic decomposition of N2O molecules.

Journal Article↗

Investigations on the local structure of Ag+/ZSM-5 catalysts and their photocatalytic reactivities for the decomposition of N2O at 298 K.

Ag+/ZSM-5 catalysts were prepared by an ion-exchange method and UV-irradiation of the catalysts in the presence of N2O led to the photocatalytic decomposition of N2O into N2 and O2 at 298 K. Investigations on the effective wavelength of irradiated UV-light for the reaction as well as the in-situ characterization of the catalysts by means of XAFS, UV-Vis, photoluminescence and FT-IR spectroscopies revealed that the photoexcitation of the Ag+-N2O complexes formed between gaseous N2O and the isolated Ag+ ions play a significant role in this reaction.

Journal Article↗

Gene expression of osteoclast differentiation factor is induced by lipopolysaccharide in mouse osteoblasts via Toll-like receptors.

Osteoclast differentiation factor (ODF), a recently identified cytokine of the TNF family, is expressed as a membrane-associated protein in osteoblasts and stromal cells. ODF stimulates the differentiation of osteoclast precursors into osteoclasts in the presence of M-CSF. Here we investigated the effects of LPS on the gene expression of ODF in mouse osteoblasts and an osteoblast cell line and found that LPS increased the ODF mRNA level. A specific inhibitor of extracellular signal-regulated kinase or protein kinase C inhibited this up-regulation, indicating that extracellular signal-regulated kinase and protein kinase C activation was involved. A protein synthesis inhibitor, cycloheximide, rather enhanced the LPS-mediated increase of ODF mRNA, and both a neutralizing Ab of TNF-alpha and a specific inhibitor of PGE synthesis failed to block the ODF mRNA increase by native LPS. Thus, LPS directly induced ODF mRNA. Mouse osteoblasts and an osteoblast cell line constitutively expressed Toll-like receptor (TLR) 2 and 4, which are known as putative LPS receptors. ODF mRNA increases in response to synthetic lipid A were defective in primary osteoblasts from C3H/HeJ mice that contain a nonfunctional mutation in the TLR4 gene, suggesting that TLR4 plays an essential role in the process. Altogether, our results indicate that ODF gene expression is directly increased in osteoblasts by LPS treatment via TLR, and this pathway may play an important role in the pathogenesis of LPS-mediated bone disorders, such as periodontitis.

Animals↗

Protective responses against experimental Mycobacterium leprae infection in mice induced by recombinant Bacillus Calmette-Guérin over-producing three putative protective antigen candidates.

The components of Ag85 (Ag85A, Ag85B, and Ag85C) are putative protective antigen candidates against mycobacterial infection. A recombinant Mycobacterium bovis Bacillus Calmette-Guérin (rBCG) over-producing Ag85A, Ag85B, and MPB51 (rBCG/BA51) was constructed. rBCG/BA51 could secrete these antigens at levels more than five times higher than parental BCG. Immunization of C57BL/6 and BALB/c mice with this rBCG reduced the multiplication of Mycobacterium leprae in the foot pads of both strains of mice. The inhibition by rBCG/BA51 was more evident than that by parental BCG.

Animals↗

Sequential change of virus markers in seroconverters with community-acquired infection of human T lymphotropic virus type I.

Twenty-three human T lymphotropic virus type I (HTLV-I) seroconverters were identified among 1120 HTLV-I-seronegative adults followed up for 11 years in an area of Japan endemic for HTLV-I. The geometric mean titer of anti-HTLV-I was 1:453 in the first year after seroconversion; the titer of each subject did not change significantly during 2-10 years of follow-up. HTLV-I proviral DNA load was quantified in 15 seroconverters, and a broad range of levels was observed-from <10 to >1000 copies/10(5) peripheral blood mononuclear cells. However, there was no obvious change in HTLV-I proviral DNA load over several years within individual subjects. Therefore, both proviral DNA load and humoral response in adult HTLV-I seroconverters were shown to stabilize within a few years after initial infection. In addition, 1 subject tested positive for HTLV-I proviral DNA before antibody seroconversion, which suggests the existence of a window period in community-acquired infection.

Aged↗

Only centromeres can supply the partition system required for ARS function in the yeast Yarrowia lipolytica.

Autonomously replicating sequences (ARSs) in the yeast Yarrowia lipolytica require two components: an origin of replication (ORI) and centromere (CEN) DNA, both of which are necessary for extrachromosomal maintenance. To investigate this cooperation in more detail, we performed a screen for genomic sequences able to confer high frequency of transformation to a plasmid-borne ORI. Our results confirm a cooperation between ORI and CEN sequences to form an ARS, since all sequences identified in this screen displayed features of centromeric DNA and included the previously characterized CEN1-1, CEN3-1 and CEN5-1 fragments. Two new centromeric DNAs were identified as they are unique, map to different chromosomes (II and IV) and induce chromosome breakage after genomic integration. A third sequence, which is adjacent to, but distinct from the previously characterized CEN1-1 region was isolated from chromosome I. Although these CEN sequences do not share significant sequence similarities, they display a complex pattern of short repeats, including conserved blocks of 9 to 14 bp and regions of dyad symmetry. Consistent with their A+T-richness and strong negative roll angle, Y. lipolytica CEN-derived sequences, but not ORIs, were capable of binding isolated Drosophila nuclear scaffolds. However, a Drosophila scaffold attachment region that functions as an ARS in other yeasts was unable to confer autonomous replication to an ORI-containing plasmid. Deletion analysis of CEN1-1 showed that the sequences responsible for the induction of chromosome breakage could be eliminated without compromising extrachromosomal maintenance. We propose that, while Y. lipolytica CEN DNA is essential for plasmid maintenance, this function can be supplied by several sub-fragments which, together, form the active chromosomal centromere. This complex organization of Y. lipolytica centromeres is reminiscent of the regional structures described in the yeast Schizosaccharomyces pombe or in multicellular eukaryotes.

Base Sequence↗

Introduction of genes encoding C4 photosynthesis enzymes into rice plants: physiological consequences.

Transgenic rice plants expressing the maize phosphoeno/pyruvate carboxylase (PEPC) and pyruvate, orthophosphate dikinase (PPDK) exhibit a higher photosynthetic capacity (up to 35%) than untransformed plants. The increased photosynthetic capacity in these plants is mainly associated with an enhanced stomatal conductance and a higher internal CO2 concentration. Plants simultaneously expressing high levels of both enzymes also have a higher photosynthetic capacity. The results suggest that both PEPC and PPDK play a key role in organic acid metabolism in the guard cells to regulate stomatal opening. Under photoinhibitory and photooxidative conditions, PEPC transgenic rice plants are capable of maintaining a higher photosynthetic rate, a higher photosynthetic quantum yield by PSII and a higher capacity to dissipate excess energy photochemically and non-photochemically than untransformed plants. Preliminary data from field trials show that relative to untransformed plants, the grain yield is about 10-20% higher in selected PEPC and 30-35% higher in PPDK transgenic rice plants, due to increased tiller number. Taken together, these results suggest that introduction of C4 photosynthesis enzymes into rice has a good potential to enhance its tolerance to stress, photosynthetic capacity and yield.

Carbon Dioxide↗

A primary culture system of rat olfactory bulb forming many synapses similar to intact ones and spontaneously generating synchronous intracellular calcium oscillations.

Previously, several studies attempting to analyze olfactory functions using dissociated culture systems of the olfactory bulb (OB) have been reported. Reciprocal dendrodendritic synapses between secondary neurons (mitral/tufted cells) and interneurons (periglomerular/granule cells) are considered to play the most important role in signal processing in the OB. However, it is unclear whether these reciprocal synapses are formed in vitro in the same way as they are in the intact OB. Thus, we synaptologically investigated the nature of cultured OB neurons. These neurons from embryonic rats were classified into four groups based on the size of their somata and their glutamic acid decarboxylase (GAD) immunoreactivity. At 14 days in vitro, most of the neurons synchronously showed spontaneous intracellular Ca2+ oscillations that were reversibly inhibited by application of D-APV and CNQX. Moreover, the frequency of the oscillations decreased and their amplitude became larger following application of bicuculline. These results suggest functional glutamatergic synaptic coupling and inhibitory GABAergic synaptic modulation. Immunocytochemical staining revealed many dot-like products (puncta) that were immunoreactive to GAD as well as to synaptophysin surrounding the cultured neurons. These results strongly indicate the presence of GABAergic synapses. The existence of synaptic contacts in OB neuron cultures was also confirmed by electron microscopy. Two types of synapses, symmetrical and asymmetrical, were morphologically recognizable. Moreover, we could also identify peculiar synapses resembling the in vivo reciprocal dendrodendritic synapses. The use of these primary culture systems will facilitate the elucidation of mechanisms underlying olfactory functions.

Animals↗

ik3-1/Cables is a substrate for cyclin-dependent kinase 3 (cdk 3).

p70ik3-1 (a 70-kDa protein) contains a cyclin box, and binds to p35cdk3 in vivo and in vitro [Matsuoka, M., Matsuura, Y., Semba, K. & Nishimoto, I. (2000) Biochem. Biophys. Res. Commun. 273, 442-447]. In spite of its structural similarity to cyclins, p70ik3-1 does not activate cyclin-dependent kinase 3 (cdk3)-mediated phosphorylation of pRb, histone H1, or the C-terminal domain of RNA polymerase II. Here, we report that Ser274 of p70ik3-1 is phosphorylated by cdk2 or cdk3 bound to cyclin A and to cyclin E in vitro. We also found that in COS7 cells in which cyclin E and cdk3 were ectopically overexpressed, the phosphorylation level of Ser274 in coexpressed p70ik3-1 is upregulated. We therefore conclude that p70ik3-1 is a substrate for cdk3-mediated phosphorylation.

Animals↗

Assessment of concordance among genealogical reconstructions from various mtDNA segments in three species of Pacific salmon (genus Oncorhynchus).

Seven segments of mitochondrial DNA (mtDNA), comprising 97% of the mitochondrial genome, were amplified by polymerase chain reaction (PCR) and examined for restriction site variation using 13 restriction endonucleases in three species of Pacific salmon: pink (Oncorhynchus gorbuscha), chum (O. keta) and sockeye (O. nerka) salmon. The distribution of variability across the seven mtDNA segments differed substantially among species. Little similarity in the distribution of variable restriction sites was found even between the mitochondrial genomes of the even- and odd-year broodlines of pink salmon. Significantly different levels of nucleotide diversity were detected among three groups of genes: six NADH-dehydrogenase genes had the highest; two rRNA genes had the lowest; and a group that included genes for ATPase and cytochrome oxidase subunits, the cytochrome b gene, and the control region had intermediate levels of nucleotide diversity. Genealogies of mtDNA haplotypes were reconstructed for each species, based on the variation in all mtDNA segments. The contributions of variation within different segments to resolution of the genealogical trees were compared within each species. With the exception of sockeye salmon, restriction site data from different genome segments tended to produce rather different trees (and hence rather different genealogies). In the majority of cases, genealogical information in different segments of mitochondrial genome was additive rather than congruent. This finding has a relevance to phylogeographic studies of other organisms and emphasizes the importance of not relying on a limited segment of the mtDNA genome to derive a phylogeographic structure.

Animals↗

Immunocytochemical study of G(i)2alpha and G(o)alpha on the epithelium surface of the rat vomeronasal organ.

To investigate in detail the distribution of G protein subtypes G(i)2alpha and G(o)alpha along the surface of the vomeronasal epithelium, we used double labeling immunocytochemical methods and electron microscopy. We examined the immunoreactivity of these surface structures with antibodies against G(i)2alpha and G(o)alpha. G(i)2alpha- and G(o)alpha-positive cells were observed at the epithelial surface and were evenly distributed. Electron microscopy revealed that strong immunoreactivities to both antibodies were observed on the microvilli and knob-like surface structures of receptor cells. No immunoreactivity was found on the microvilli or surface membranes of supporting cells. This expression pattern is similar to that reported for putative pheromone receptors. These data confirm that there are two distinct classes of vomeronasal receptor cells expressed at the surface of the epithelium. These two classes of receptors correspond to the same G(i)2alpha- and G(o)alpha-positive cells distributed in cell body layers of the epithelium and in the axon terminals in the accessory olfactory bulb.

Animals↗

The mouse putative pheromone receptor was specifically activated by stimulation with male mouse urine.

To detect the biological activity of mammalian putative pheromone receptors (V1Rs and V2Rs), the mouse V1R gene was introduced into a primary culture of vomeronasal cells using the adenovirus expression system, and the response of these cells to mouse urine was analyzed by calcium imaging. These cells specifically responded to male but not female mouse urine. This response was attenuated by pertussis toxin, a specific inhibitor of G-protein G(ialpha)/G(oalpha) coupling from receptors. Our findings indicate that a putative pheromone receptor was specifically activated by mouse urine, a major source of mouse pheromones, and suggest that G(i)/G(o) are functionally coupled with the receptor.

Amino Acid Sequence↗

High level expression of C4-specific NADP-malic enzyme in leaves and impairment of photoautotrophic growth in a C3 plant, rice.

The chloroplastic NADP-malic enzyme (NADP-ME) is a key enzyme of the C4 photosynthesis pathway in NADP-ME type C4 plants such as maize. To express the chloroplastic NADP-ME in leaves of a C3 plant, rice, full-length cDNAs encoding the rice C3-specific isoform and the maize C4-specific isoform of the enzyme were expressed under the control of the rice CAB: promoter. Transformants carrying the rice cDNA showed the NADP-ME activities in the leaves less than several-fold that of non-transformants, while those carrying the maize cDNA showed activities up to 30-fold that of non-transformants or about 60% of the NADP-ME activity of maize leaves. These results indicate that expression of the rice C3-specific NADP-ME is suppressed at co- and/or post-transcriptional levels by some regulation mechanisms intrinsic to rice, while that of the foreign C4-specific isoform can escape from such suppression. The accumulation of the maize C4-specific NADP-ME led to bleaching of leaf color and growth hindrance in rice plants under natural light. These deteriorative effects resulted from enhanced photoinhibition of photosynthesis due to an increase in the level of NADPH inside the chloroplast by the action of the maize enzyme.

Gene Expression↗

Expression of a gibberellin 2-oxidase gene around the shoot apex is related to phase transition in rice.

A major catabolic pathway for gibberellin (GA) is initiated by 2beta-hydroxylation, a reaction catalyzed by GA 2-oxidase. We have isolated and characterized a cDNA, designated Oryza sativa GA 2-oxidase 1 (OsGA2ox1) from rice (Oryza sativa L. cv Nipponbare) that encodes a GA 2-oxidase. The encoded protein, produced by heterologous expression in Escherichia coli, converted GA(1), GA(4), GA(9), GA(20), and GA(44) to the corresponding 2beta-hydroxylated products GA(8), GA(34), GA(51), GA(29), and GA(98), respectively. Ectopic expression of the OsGA2ox1 cDNA in transgenic rice inhibited stem elongation and the development of reproductive organs. These transgenic plants were deficient in endogenous GA(1). These results indicate that OsGA2ox1 encodes a GA 2-oxidase, which is functional not only in vitro but also in vivo. OsGA2ox1 was expressed in shoot apex and roots but not in leaves and stems. In situ hybridization analysis revealed that OsGA2ox1 mRNA was localized in a ring at the basal region of leaf primordia and young leaves. This ring-shaped expression around the shoot apex was drastically decreased after the phase transition from vegetative to reproductive growth. It was absent in the floral meristem, but it was still present in the lateral meristem that remained in the vegetative phase. These observations suggest that OsGA2ox1 controls the level of bioactive GAs in the shoot apical meristem; therefore, reduction in its expression may contribute to the early development of the inflorescence meristem.

Amino Acid Sequence↗