ZEITLUPE and FKF1: novel connections between flowering time and circadian clock control.
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Arabidopsis Proteins↗
Biomedical subjects
T Mizoguchi
Publications and source records attributed to T Mizoguchi.
Surgical effects of combined trabeculotomy and sinusotomy compared to trabeculotomy alone.
PURPOSE: To elucidate long-term surgical results and problems of combined trabeculotomy and sinusotomy in comparison with trabeculotomy alone. METHODS: The study included 91 glaucomatous eyes in 57 patients with primary open angle glaucoma, who underwent either combined trabeculotomy and sinusotomy (40 eyes) or trabeculotomy alone (51 eyes). The mean follow-up period (+/-standard deviation) was 46.1+/-10.4 months (range: 30 to 60 months). RESULTS: At the final examination, in 37 (93%) of the 40 eyes after combined trabeculotomy and sinusotomy and in 40 (78%) of the 51 eyes after trabeculotomy alone, intraocular pressures were well controlled. The mean intraocular pressures at the end of the first postoperative year were 15.6+/-3.0 mmHg in the 40 eyes after the combined procedure, and 17.8+/-3.1 mmHg in the 51 eyes after trabeculotomy alone (p=0.0001, unpaired t-test). Complications included Descemet's membrane detachment in 1 eye (3%) and 4 eyes (8%), and transient elevation of intraocular pressure to higher than 30 mmHg (IOP spike) in 6 eyes (15%) and 11 eyes (22%), respectively, after the combined procedure and trabeculotomy alone. CONCLUSION: Combined trabeculotomy and sinusotomy is a useful surgical modality for the treatment of primary open angle glaucoma to obtain lower intraocular pressure levels than those after trabeculotomy alone.
Female↗
Various abiotic stresses rapidly activate Arabidopsis MAP kinases ATMPK4 and ATMPK6.
Mitogen-activated protein kinase (MAP kinase, MAPK) cascades play pivotal roles in signal transduction of extracellular stimuli, such as environmental stresses and growth regulators, in various organisms. Arabidopsis thaliana MAP kinases constitute a gene family, but stimulatory signals for each MAP kinase have not been elucidated. Here we show that environmental stresses such as low temperature, low humidity, hyper-osmolarity, touch and wounding induce rapid and transient activation of the Arabidopsis MAP kinases ATMPK4 and ATMPK6. Activation of ATMPK4 and ATMPK6 was associated with tyrosine phosphorylation but not with the amounts of mRNA or protein. Kinetics during activation differ between these two MAP kinases. These results suggest that ATMPK4 and ATMPK6 are involved in distinct signal transduction pathways responding to these environmental stresses.
Abscisic Acid↗
Immobilization of ligand-modified polyamidoamine dendrimer for cultivation of hepatoma cells.
Cationic polyamidoamine dendrimers are known to be highly branched cascade polymers. The core part of these polymers, tris(2-aminoethyl)amine, was immobilized onto polystyrene plates to which animal cells do not adhere. using photoreactive 4-(3-trifluoromethylazirino) benzoyl-N-succinimide (TDBA-OSu). Cells of a rat hepatoma cell line, H4-II-E-C3, adhered to a surface immobilized with a first-generation dendrimer probably through interactions between the terminal amino groups of the dendrimer and the cell membranes. The adhered cells were viable, could proliferate, and exhibited urea synthetic activity. The modification of the terminal amino groups with fructose increased the final number of cells obtained after 5 days of cultivation. Multigeneration dendrimers were prepared by repeated linkage of tris(2-aminoethyl)amine with the amino groups. Theoretically, the number of terminal amino groups available for ligand modification is twice as much for each generation of dendrimer growth. Cells cultivated on multigeneration fructose-modified dendrimers exhibited enhanced urea synthetic activity. The use of ligand-modified dendrimers is, therefore, considered to be very promising for the construction of bioartificial organs based on cultivation of the animal cells.
Animals↗
Structural transformation among the aggregate forms of bacteriochlorophyll c as determined by electronic-absorption and NMR spectroscopies: dependence on the stereoisomeric configuration and on the bulkiness of the 8-C side chain.
Transformation among the aggregate forms of bacteriochlorophyll (BChl) c characterized by the wavelength of the Qy absorption, i.e. the dimer (B675), B705, B720 and B745, was traced by electronic-absorption spectroscopy for each of the isomers including R[E,E], R[P,E], R[I,E], S[P,E] and S[I,E] suspended in the mixtures of methylene chloride and n-hexane. A combination of NMR spectroscopy determining the structural motifs and calculation of the shift of the Qy absorption reflecting the long-range transition dipole-transition dipole interactions among the macrocycles in the entire aggregate structures proposed the following models: B705d (B705d'), a linear array of straight (inclined) columns consisting of a pair of the piggyback dimers; B720d and B745d, an assembly of two and five shifted-inclined columns consisting of more than six piggyback dimers; and B720m and B745m, an assembly of one and two parallel stepwise stacking of approximately 30 monomers. Calculations of the steric energies rationalized two different pathways of transformations: the dimer-->B705d (B705d')-->B720d-->B745d for the R isomers; and the monomer-->(B720m)-->B745m for the S isomers. Addition of S[I,E] seems to trigger the B745d-->B745m transformation of the R isomers.
Bacterial Proteins↗
p125 is a novel mammalian Sec23p-interacting protein with structural similarity to phospholipid-modifying proteins.
COPII-coated vesicles are involved in protein transport from the endoplasmic reticulum to the Golgi apparatus. COPII consists of three parts: Sar1p and the two protein complexes, Sec23p-Sec24p and Sec13p-Sec31p. Using a glutathione S-transferase fusion protein with mouse Sec23p, we identified a novel mammalian Sec23p-interacting protein, p125, which is clearly distinct from Sec24p. The N-terminal region of p125 is rich in proline residues, and the central and C-terminal regions exhibit significant homology to phospholipid-modifying proteins, especially phosphatidic acid preferring-phospholipase A1. We transiently expressed p125 and mouse Sec23p in mammalian cells and examined their interaction. The results showed that the N-terminal region of p125 is important for the interaction with Sec23p. We confirmed the interaction between the two proteins by a yeast two-hybrid assay. Overexpression of p125, like that of mammalian Sec23p, caused disorganization of the endoplasmic reticulum-Golgi intermediate compartment and Golgi apparatus, suggesting its role in the early secretory pathway.
Amino Acid Sequence↗
Repression of nitrogenase by ethanol in nitrogen-deprived cultures of Rhodovulum sulfidophilum.
Light-dependent H2 evolution did not occur in nitrogen-deprived cultures of Rhodovulum sulfidophilum in the presence of ethanol. When ethanol was added to cells which had been grown with ammonia, derepression of the nitrogen fixation genes (nifHD) was inhibited at an ethanol concentration of 1 mM. On the other hand, when cells had nitrogenase-catalyzed proton-reducing activity prior to ethanol addition, reduction of the nifHD transcript level did not occur after the addition. In cells grown with ammonia, concomitant addition of an auxiliary oxidant such as dimethylsulfoxide or sodium bicarbonate resulted in derepression of nitrogenase activity in the presence of ethanol. These results suggest that the electron-accepting process is necessary for derepression of nif genes in cultures which use ethanol as the electron donor.
Ammonia↗
The structure of an aggregate form of bacteriochlorophyll c showing the Qy absorption at 705 nm as determined by the ring-current effects on 1H and 13C nuclei and by 1H-1H intermolecular NOE correlations.
13C-enriched bacteriochlorophyll c (R[E, E] BChl cF) was suspended in chloroform to form an aggregate showing the Qy absorption at 705 nm. (1) The aggregate exhibited several largely split 13C-NMR signals suggesting the presence of non-equivalent BChl c molecules in the form of the piggyback dimer. (2) Changes in the 13C chemical shifts were traced when methanol was titrated to dissolve the aggregate, and the aggregation shifts (in reference to the monomeric state) were determined as a function of the amount of methanol titrated, and they were analyzed empirically. (3) The ring-current effects were calculated based on the loop-current approximation, and the results were compared with the observed aggregation shifts for 13C and 1H nuclei (the 1H aggregation shifts were determined by extrapolation of the data taken from Mizoguchi, T.; Limantara, L.; Matsuura, K.; Shimada, K.; Koyama, Y. J Mol Structure 1996, 379, 249-265). The results showed that the assembly of two straight columns consisting of the piggyback dimer stacked in the antiparallel orientation is the best choice as a model for the B705 aggregate. (4) Three-dimensional F1 13C-edited F3 13C-filtered heteronuclear single-quantum nuclear-Overhauser-effect spectroscopy was applied to the aggregate consisting of a 1:1 mixture of 13C-labeled and unlabeled BChl c in order to selectively detect the intermolecular 1H-1H NOE correlations. The NOE correlations were explained in terms of a straight column, supporting the above model.
Bacterial Proteins↗
Formation of lens aldose reductase mixed disulfides with GSH by UV irradiation and its proteolysis by lens calpain.
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Aldehyde Reductase↗
Effectiveness of polyamidoamine dendrimers modified with tripeptide growth factor, glycyl-L-histidyl-L-lysine, for enhancement of function of hepatoma cells.
Cationic polyamidoamine dendrimers are known to be highly branched cascade polymers. Tripeptide growth factor, glycyl-L-histidyl-L-lysine (GHK), was employed as a ligand for activation or attachment of cells from a rat hepatoma cell line, H4-H-E-C3, and immobilized at the terminus of the dendrimer (GHK-dendrimer) to develop a suitable surface for use as a culture substratum in the bioartificial liver support system (BAL). The growth of cells was inhibited by increasing the number of generations of GHK-dendrimers. On the other hand, urea synthesis and lidocaine clearance of the cells adhered on fifth generation GHK-dendrimers were enhanced much more than on first generation GHK-dendrimers. GHK was shown to act as a growth inhibitor and an activator of hepatoma cells. These properties of GHK are advantageous for the utilization of hepatoma cells in BAL. Ligand-modified dendrimers are very promising for the creation of a high-performance substratum for cell culture and high performance bioartificial organs, as well as for high-performance bioartificial liver systems. GHK may have the potential to be a highly useful ligand.
Journal Article↗
Increase in thioredoxin activity of intestinal epithelial cells mediated by oxidative stress.
Hydrogen peroxide was cytotoxic to the small intestine epithelial cell line, IEC-6, as judged from an MTT assay and the release of lactate dehydrogenase. The glutathione S-transferase and thioredoxin reductase activities and SH content decreased dose-dependently with H2O2, but thioredoxin activity increased at low H2O2 concentrations. In addition, the increase in thioredoxin activity was time-dependent during the initial stages of oxidative stress. A reverse transcription-polymerase chain reaction (RT-PCR) amplification also showed that the mRNA content in IEC-6 cells increased time-dependently at 0.25 mM H2O2. These results indicate that cellular oxidative shock causes an increase in the activity of thioredoxin, which is involved in the defense mechanism against oxidative stress.
Animals↗
Immobilization of tripeptide growth factor glycyl-L-histidyl-L-lysine on poly(vinylalcohol)-quarternized stilbazole (PVA-SbQ) and its use as a ligand for hepatocyte attachment.
A tripeptide growth factor, glycyl-L-histidyl-L-lysine (GHK), was immobilized on the surface of poly(vinylalcohol)-quarternized stilbazole (PVA-SbQ) gel. The photoreactive substance, 4-(3-trifluoromethylazirino)benzoyl-N-hydroxysuccinimide (TDBA-OSu), was employed to link the gel and ligand GHK. The density of immobilized GHK was 70 nmol/cm2. Isolated rat hepatocytes were inoculated on the GHK-immobilized PVA-SbQ gel and cultured for 5 d. About 24 h after inoculation, hepatocytes started to aggregate and formed multicellular spheroids while almost no cells attached to GHK-non-immobilized PVA-SbQ gel. The formed spheroids attached firmly to the surface of PVA-SbQ gel for 5 d. GHK was, thus, shown to be an effective ligand for hepatocyte attachment. Dodecamethylenediamine was used to extend the length between the gel surface and GHK. Extension of the length significantly increased the number of attached hepatocytes.
Animals↗
cDNA sequence of bovine thioredoxin.
In this paper, we report the cDNA sequence of bovine thioredoxin. We determined the full-length cDNA sequence of bovine thioredoxin by RT-PCR, 5'-RACE and 3'-RACE methods. Currently, the thioredoxin cDNA sequences of only five mammalian species (human, macaca, mouse, ovine and rat) are registered in the GenBank database. We performed sequence comparisons on the total cDNA sequence and the coding region, and produced a multialignment between the amino acid sequences of bovine and other mammalian thioredoxins. The amino acid sequences of thioredoxins are highly conserved among mammalian species, for example, only one difference exists between the amino acid sequences of bovine and ovine thioredoxin.
Animals↗
cDNA sequence of bovine thioredoxin.
In this paper, we report the cDNA sequence of bovine thioredoxin. We determined the full-length cDNA sequence of bovine thioredoxin by RT-PCR, 5'-RACE and 3'-RACE methods. Currently, the thioredoxin cDNA sequences of only five mammalian species (human, macaca, mouse, ovine and rat) are registered in the GenBank database. We performed sequence comparisons on the total cDNA sequence and the coding region, and produced a multialignment between the amino acid sequences of bovine and other mammalian thioredoxins. The amino acid sequences of thioredoxins are highly conserved among mammalian species, for example, only one difference exists between the amino acid sequences of bovine and ovine thioredoxin.
Amino Acid Sequence↗
Intraoperative transesophageal echocardiography for evaluation of mitral valve repair.
BACKGROUND AND AIM OF THE STUDY: Although mitral valve repair is a well-established procedure, incorrect assessment of the repaired valve may occasionally lead to the need for reoperation. This study was performed to evaluate the accuracy of color Doppler in assessing the competence of the repaired mitral valve. METHODS: Transesophageal echocardiography (TEE) and left ventriculography were each performed in 72 patients to compare the two techniques and a semi-quantitative index derived. Using this relationship, post bypass intraoperative TEE was then performed in 34 patients who underwent mitral valve repair, in order to assess the competence of the repaired valve. RESULTS: Significant differences were apparent in maximal regurgitant mosaic area between angiographic grade 0, and grades 1+ (p = 0.0006), 1+ and 2+ (p < 0.0001) and 2+ and 3+ (p = 0.0010). A maximal regurgitant area < 2 cm2 predicted angiographic grade as 0 (sensitivity 100%, specificity 95%), an area of 2-4 cm2 as 1+ (sensitivity 82%, specificity 100%), an area of 4-7 cm2 as 2+ (sensitivity 78%, specificity 90%), and an area > 7 cm2 as grade 3+ or 4+ (sensitivity 79%, specificity 93%). All 34 patients completed valve repair with the maximal regurgitant mosaic area < 2.5 cm2. Postoperative left ventriculography showed grade +1 in only five patients; four of these completed mitral valve repair with a maximal mosaic area > 2.0 cm2 as assessed by post bypass intraoperative TEE. During follow up, transthoracic echocardiography (TTE) detected recurrent mitral regurgitation which required mitral valve replacement in one patient, and rapid progression of mitral regurgitation in three patients. CONCLUSIONS: It is important that mitral valve repair should be completed with a maximal mosaic area < 2.0 cm2 as assessed by intraoperative TEE, in order to reduce the need for reoperation.
Adult↗
Molecular cloning and characterization of three cDNAs encoding putative mitogen-activated protein kinase kinases (MAPKKs) in Arabidopsis thaliana.
We isolated three Arabidopsis thaliana cDNA clones (ATMKK3, ATMKK4 and ATMKK5) encoding protein kinases with extensive homology to the mitogen-activated protein kinase kinases (MAPKKs) of various organisms in the catalytic domain. ATMKK3 shows high homology (85% identity) to NPK2, a tobacco MAPKK homologue. ATMKK4 and 5 are closely related to each other (84% identity). Phylogenetic analysis showed that the plant MAPKKs constitute at least three subgroups. The recombinant ATMKK3 and ATMKK4 were expressed as a fusion protein with glutathione S-transferase (GST) in Escherichia coli. Affinity purified GST-ATMKK3 and GST-ATMKK4 proteins contained phosphorylation activity, which shows that both the ATMKK3 and ATMKK4 genes encode functional protein kinases. Northern blot analysis revealed that the ATMKK3 gene expressed in all the organs. The levels of ATMKK4 and 5 mRNAs were relatively higher in steins and leaves than in flowers and roots. We determined the map positions of the ATMKK3, 4 and 5 genes on Arabidopsis chromosomes by RFLP mapping using P1 genomic clones.
Amino Acid Sequence↗
Isolation of ATMEKK1 (a MAP kinase kinase kinase)-interacting proteins and analysis of a MAP kinase cascade in Arabidopsis.
In plants, a number of MAP kinase (MAPK), MAPK kinase (MAPKK), and MAPKK kinase (MAPKKK) homologues have been reported. However, there have been no reports of protein-protein interactions between these kinases or molecular analysis of MAPK cascades in higher plants. To analyze a possible MAPK cascade in Arabidopsis thaliana, we took two molecular approaches. One is the two-hybrid screening of ATMEKK1 (a MAPKKK)-interacting proteins; the other is an analysis of physical and functional interactions among isolated MAPK, MAPKK, and MAPKKK homologues from Arabidopsis. In two-hybrid screening using ATMEKK1 as bait, we isolated a novel MAPKK homologue, ATMKK2, a MAPK homologue, ATMPK4, and an unknown protein. ATMKK2 has high sequence similarity with MEK1 (a MAPKK) in Arabidopsis. Based on yeast two-hybrid analysis, we detected protein-protein interactions between ATMEKK1 and ATMKK2/MEK1 (MAPKKs), between ATMKK2/MEK1 and ATMPK4 (a MAPK), and between ATMPK4 and ATMEKK1. ATMPK4 and ATMKK2/MEK1 interacted with two distinct regions of ATMEKK1, the N-terminal regulatory domain and the C-terminal kinase domain, respectively. Coexpression of ATMEKK1 increased the ability of two closely related MAPKKs, ATMKK2 and MEK1, to complement a growth defect of the yeast pbs2 mutant. Coexpression of ATMPK4 and MEK1 complemented a growth defect of the yeast mpk1 and bck1 mutants. By contrast, other combinations of MAPKs and MAPKKs did not suppress these yeast mutations. These results suggest that ATMEKK1, ATMKK2/MEK1, and ATMPK4 may constitute a MAP kinase cascade.
Alanine↗
Identification of a possible MAP kinase cascade in Arabidopsis thaliana based on pairwise yeast two-hybrid analysis and functional complementation tests of yeast mutants.
A possible MAP kinase (MAPK) cascade of Arabidopsis thaliana was identified on the basis of both yeast 2-hybrid analysis and complementation analysis of yeast mutants. Specific protein-protein interactions between ATMPK4 (a MAPK) and MEK1 (a MAPKK) and interactions between MEK1 and ATMEKK1 (a MAPKKK) were detected by using the 2-hybrid system. A growth defect of the yeast mpk1delta mutant was reversed by coexpression of ATMPK4 and MEK1. Coexpression of the N-terminal deletion form of ATMEKK1 increased the ability of MEK1 to suppress a growth defect of the yeast pbs2delta mutant. These results suggest that ATMPK4, MEK1, and ATMEKK1 may interact with each other and constitute a specific MAPK cascade in Arabidopsis. This is the first demonstration of a possible MAPK cascade in plants.
Arabidopsis↗