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T Kubo

Publications and source records attributed to T Kubo.

At least 487 records · Page 27Linked to original sources
PubMed1995

Effects of MK801 on Fos expression in the rat brainstem after unilateral labyrinthectomy.

Unilateral labyrinthectomy (UL) causes ocular and postural asymmetries, which disappear over time in the processes of equilibrium recovery known as vestibular compensation. It has been reported that N-methyl-D-aspartate (NMDA) receptors are involved in vestibular compensation. In the present study, in order to elucidate the NMDA receptor-mediated neural circuit responsible for the development of vestibular compensation, we used Fos expression as a marker of neural activation and examined the effects of MK801, a specific antagonist of NMDA receptors, on UL-induced Fos expression in the rat brainstem. After UL, Fos-like immunoreactive (-LIR) neurons were observed in the ipsilateral medial vestibular nucleus (ipsi-MVe), the contralateral prepositus hypoglossal nucleus (contra-PrH) and the contralateral inferior olive beta subnucleus (contra-IOb). Fos-LIR neurons gradually disappeared in the processes of vestibular compensation. It is suggested that the activation of the ipsi-MVe, the contra-PrH and the contra-IOb neurons after UL are the initial event of vestibular compensation. Intraperitoneal injection of MK801 in the processes of vestibular compensation caused reappearance of UL-induced behavioral deficits. During the decompensation induced by MK801, Fos-LIR neurons appeared in the contra-MVe, the ipsi-PrH and the bilateral-IOB. It is suggested that the contra-MVe, the ipsi-PrH and the bilateral-IOb neurons are inhibited by glutamatergic synapses driving inhibitory neurons via NMDA receptors in the processes of vestibular compensation and that disinhibition of these nuclei induced by MK801 causes decompensation. However, MK801 caused neither Fos expression nor behavioral decompensation after vestibular compensation is accomplished. All these findings that the NMDA receptor-mediated inhibitory modulation in the central vestibular system plays an important role for the initial processes of the development of vestibular compensation.

Amino Acid Sequence↗
PubMed1995

DNA topoisomerase II alpha gene expression under transcriptional control in etoposide/teniposide-resistant human cancer cells.

We previously isolated etoposide/teniposide-resistant cell lines from human cancer KB cells, designated KB/VP-2 and KB/VM-4, respectively, and we found that decreased expression of topoisomerase II alpha was associated with the acquisition of etoposide/teniposide resistance in both resistant cell lines. In this study, we studied how the expression of the DNA topoisomerase II alpha gene is regulated in drug-resistant cell lines at the transcriptional level. We first examined whether the decreased topoisomerase II alpha mRNA level was due to a shorter lifetime of mRNA molecules in drug-resistant cell lines. A comparison of the degradation kinetics of topoisomerase II alpha mRNA demonstrated that there was no difference in mRNA stability between both resistant cell lines and their parental counterpart. A run-on experiment with isolated nuclei showed that the transcriptional activity of topoisomerase II alpha gene of both resistant cell lines constituted less than 20% of the parental KB cells. The activity of DNA topoisomerase II alpha promoter in resistant cells was also less than 20% of that in KB cells when transient transfection assays were performed with the promoter-driven bacterial chloramphenicol acetyltransferase gene. Among the several transcription factors that might be involved in DNA topoisomerase II alpha gene expression, expression of Sp3, an inhibitory member of the Sp1 family, was elevated to about 3-fold higher in both resistant cell lines than their parental counterpart. These results indicated that the expression of DNA topoisomerase II alpha gene decreased at the transcriptional level through the enhanced expression of Sp3 in our two etoposide/teniposide-resistant cell lines.

Antigens, Neoplasm↗
PubMed1995

Enhanced release of acetylcholine in the rostral ventrolateral medulla of spontaneously hypertensive rats.

We examined whether the altered rostral ventrolateral medulla (RVLM) cholinergic function in spontaneously hypertensive rats (SHR) results from enhanced presynaptic cholinergic tone. Male 12- to 16-week-old SHR and age-matched Wistar Kyoto rats (WKY) were anesthetized, paralyzed and artificially ventilated. Unilateral microinjection of cholinergic agents into the RVLM produced a pressor response. The pressor response to physostigmine was greater in SHR than that of WKY whereas the response to ACh and carbachol was the same in WKY and SHR. Bilateral microinjection of scopolamine produced a decrease in blood pressure. The depressor response was greater in SHR than that of WKY. When a microdialysis probe was placed in the RVLM, ACh release in the RVLM was greater in SHR than that of WKY. Choline acetyltransferase (CAT) activity was increased only in the rostro-ventral part of the medulla, which contained the RVLM, but not in other parts of the medulla oblongata. Physostigmine (0.5 mg/kg, i.p.)-induced increases in ACh content were also enhanced only in the rostro-ventral part of the medulla. These results provide direct evidence that ACh release in the RVLM is enhanced in SHR. It appears that the enhanced cholinergic activity in the RVLM of SHR results from an increase in cholinergic impulse flow in the RVLM of SHR. This abnormality may play a role in the maintenance of hypertension in SHR.

Acetylcholine↗
PubMed1995

Molecular cloning and nuclear localization of ATBP, a novel (A+T)-stretch-binding protein of Sarcophaga peregrina (flesh fly).

Previously, we purified a DNA-binding protein, named ATBP, from cultured Sarcophaga cells. This protein was found to bind specifically to (A+T) stretches that locate in the 5'-upstream region and the intron of the Sarcophaga lectin gene [Matsui, M., Kobayashi, A., Kubo, T. & Natori, S. (1994) Eur. J. Biochem. 219, 449-454]. Here, we report molecular cloning of a cDNA for ATBP. The cDNA encoded a protein with two zinc-finger motifs, an acidic domain, two glutamine-rich domains and a glutamine run. Immunofluorescence study revealed that ATBP was localized in the nuclei. Thus, ATBP is possibly a transcription factor for genes having (A+T) stretches in their promoter and/or enhancer regions.

Amino Acid Sequence↗
PubMed1995

Brain-derived neurotrophic factor (BDNF) can prevent apoptosis of rat cerebellar granule neurons in culture.

Cerebellar granule neurons obtained from 9-day-old rats were grown in vitro for 4 days in high K+ (26 mM) medium. The culture medium was then replaced with that containing low K+ (5 mM) which caused a large number of granule neurons to die. The death of granule neurons has been characterized as apoptosis. In this study, we investigated the effects of various neurotrophins on neuronal survival using the above system. We found that brain-derived neurotrophic factor (BDNF) and neurotrophin-4/5 (NT-4/5) but not nerve growth factor (NGF) can protect these neurons from apoptosis in low K+. Neurotrophin-3 (NT-3) had a small effect on neuronal survival as reported. To determine whether the granule neurons respond directly to BDNF, we analyzed the induction of the Fos protein in these neurons. Individual cells that synthesize Fos protein after exposure to neurotrophin can be recognized using antibodies to Fos. Immunocytochemical staining of the cultures demonstrated that a relatively large number of cerebellar granule neurons showed immunoreactivity in response to BDNF, but few of them were immunoreactive in the absence of BDNF or in the presence of NGF. Our results suggested that BDNF has a direct effect on mature cerebellar granule neurons and can protect these neurons from apoptosis in low K+.

Animals↗
PubMed1995

Ca(2+)-independent fusion of secretory granules with phospholipase A2-treated plasma membranes in vitro.

The fusion of secretory granules with plasma membranes prepared from rat parotid gland was studied in vitro to clarify the mechanism of exocytosis. Fusion of the granules with plasma membranes was measured by a fluorescence-dequenching assay with octadecyl rhodamine B, and release of amylase was also measured to confirm the fusion as a final step of the secretory process. Plasma membranes that had been pretreated with porcine phospholipase A2 (PLA2) in the presence of 20 microM Ca2+ fused with the granules within 30 s, and induced amylase release by reacting with the membranes of granules, whereas without this pretreatment they had no significant effect. The fusion process accompanied by amylase release was induced in the presence of 10 mM EGTA, and therefore was apparently Ca(2+)-independent. On the other hand, the presence of EGTA or 100 microM quinacrine, an inhibitor of PLA2, during treatment of plasma membranes with PLA2 inhibited their fusogenic activity, suggesting the importance of activation of PLA2. Arachidonic acid and linoleic acid were released from the plasma membranes during the PLA2 treatment. The presence of albumin, an adsorbent of fatty acids, during the treatment also inhibited the activity. Pretreatment of the membranes with arachidonic acid or linoleic acid did not have any effect, but the presence of exogenously added arachidonic acid during PLA2 treatment enhanced the membrane-fusion-inducing effect of PLA2. Pretreatment of the membranes with lysophosphatidylcholine induced fusogenic activity. These findings suggest that the conformational change in the plasma-membrane phospholipids induced by PLA2 and the presence of arachidonic acid or linoleic acid produced by PLA2 are important in the process of fusion of secretory granules with the plasma membranes of rat parotid acinar cells and that the fusion process itself is independent of Ca2+.

5,8,11,14-Eicosatetraynoic Acid↗
PubMed1995

Structure-function relationship of yeast S-II in terms of stimulation of RNA polymerase II, arrest relief, and suppression of 6-azauracil sensitivity.

The yeast S-II null mutant is viable, but the mutation induces sensitivity to 6-azauracil. To examine whether the region needed for stimulation of RNA polymerase II and that for suppression of 6-azauracil sensitivity in the S-II molecule could be separated, we constructed various deletion mutants of S-II and expressed them in the null mutant using the GAL1 promoter to see if the mutant proteins suppressed 6-azauracil sensitivity. We also expressed these constructs in Escherichia coli, purified the mutant proteins to homogeneity, and examined if they stimulated RNA polymerase II. We found that a mutant protein lacking the first 147 amino acid residues suppressed 6-azauracil sensitivity but that removal of 2 additional residues completely abolished the suppression. A mutant protein lacking the first 141 residues had activity to stimulate RNA polymerase II, whereas removal of 10 additional residues completely abolished this activity. We also examined arrest-relief activity of these mutant proteins and found that there is a good correlation between RNA polymerase II-stimulating activity and arrest-relief activity. Therefore, at least the last 168 residues of S-II are sufficient for expressing these three activities.

Antimetabolites↗
PubMed1995

Molecular cloning and sequencing of arylphorin-binding protein in protein granules of the Sarcophaga fat body. Implications of a post-translational processing mechanism.

Previously, we identified an arylphorin-binding protein of Sarcophaga peregrina (flesh fly) with a molecular mass of 120 kDa and suggested its participation in the selective uptake of arylphorin from the hemolymph into the pupal fat body at metamorphosis (Ueno, K., and Natori, S. (1984) J. Biol. Chem. 259, 12107-12111). This paper reports the isolation and sequencing of cDNA for the 120-kDa protein. This protein consists of 1146 amino acid residues. Immunoblotting and RNA blotting experiments revealed that this protein is present as two fragments of 76 kDa (695 residues) and 53 kDa (451 residues) in the larval fat body. When larvae pupate, the 120-kDa protein gene is further activated and the complete 120-kDa protein is synthesized without fragmentation. This suggests a novel mechanism for the production of the 120-kDa protein regulated by a proteinase depending upon the stage of development of Sarcophaga. All of these proteins were found to be localized in protein granules in the adipocytes.

Amino Acid Sequence↗
PubMed1995

Mid-trimester fetal sex determination from maternal peripheral blood by fluorescence in situ hybridization without enrichment of fetal cells.

To determine the fetal sex on 30 women who were 16-20 weeks pregnant, about 100,000 maternal blood nucleated cells were analysed by means of fluorescence in situ hybridization (FISH) with a Y-chromosome-specific DNA probe. Cells with the hybridization signal were detected in 12 of the 30 women. All the 12 mothers gave birth to a male child. Of the other 18 women who had no Y-positive cells in the peripheral blood, 14 gave birth to a female child and four gave birth to a male child. These false-negative results probably occurred because the number of cells examined was inadequate. The data obtained in this study suggest that fetal sex determination using maternal peripheral blood with FISH is possible and that this diagnostic method will be clinically useful when more cells are analysed.

Blood Cells↗
PubMed1995

Exercise at high temperature causes maternal hyperthermia and fetal anomalies in rats.

Hyperthermia is thought to be a teratogen in many animal species and also in humans. It has been reported that hyperthermia caused by sauna, hot tub, or fever during the early stages of pregnancy is related to an increased risk for neural tube defects. During exercise, especially in hot conditions, body temperature can also rise to fairly high levels. Thus, we can surmise that hyperthermia induced by exercise can also cause fetal malformation. To investigate this hypothesis, pregnant rats at 9 days of gestation were divided into four groups. In the first group, the animals were made to swim for 30 minutes in water at a temperature of 40.5 degrees C. In the second group, they were restrained and immersed in water for the same time at the same temperature. In the third group, the rats were forced to swim in water at 36.0 degrees C. The fourth group were controls. The core temperature of the rats was measured during these procedures. On the 18th gestational day, fetuses were extracted by cesarean section. The elevation of maternal core temperature was significantly greater in the first group than in the other groups. In the first group, 69% of fetuses had various external anomalies. No anomalies were found in the other groups. Our results show that exercise in hot conditions caused the elevation of core temperature and resulted in fetal anomalies in rats.

Animals↗
PubMed1995

Physical and gene organization of mitochondrial DNA from the fertile cytoplasm of sugarbeet (Beta vulgaris L.).

We have constructed a complete physical map of the mitochondrial genome from the male-fertile cytoplasm of sugarbeet. The entire sequence complexity can be represented on a single circular master chromosome of 358 kb. This master chromosome contains three copies of one recombinationally active repeat sequence, with two copies in direct orientation and the other in inverted orientation. The positions of the rRNA genes and of 23 polypeptide genes, determined by filter hybridization, are scattered throughout the genome, with triplicate rrn26 genes located partially or entirely within the recombination-repeat elements. Three ribosomal-protein genes (rps1A, rps14 and rps19) were found to be absent from sugarbeet mtDNA. Our results also reveal that at least six regions homologous with cDNA are dispersed in the mitochondrial genome.

Chromosome Mapping↗
PubMed1995

The chloroplast trnP-trnW-petG gene cluster in the mitochondrial genomes of Beta vulgaris, B. trigyna and B. webbiana: evolutionary aspects.

The chloroplast trnP-trnW-petG gene cluster has been identified in the mitochondrial DNA (mtDNA) of sugar beet (Beta vulgaris). The chloroplast-derived trnW gene is transcribed in the mitochondria; the other two genes, however, do not seem to be transcribed. This gene cluster is also present in the mitochondrial genomes of two wild Beta species, B. trigyna and B. webbiana. Sugar beet and the two wild relatives share 100% sequence identity in the coding regions of both the mitochondrial trnP and trnW genes. On the other hand, the petG genes from the wild Beta mtDNAs were found to be disrupted either by a 5-bp duplication (B. trigyna) or by a deletion of the 5' region (B. webbiana). A data-base search revealed that a conserved sequence of 60 bp is present in the trnP-trnW intergenic region of the mitochondrial genomes of the three Beta species as well as in other higher plants, including wheat and maize, and that the conserved sequence is absent from the chloroplast counterpart. Our results thus favour the hypothesis of a monophyletic origin of the trnP-trnW-petG cluster found in the plant mitochondrial genomes examined.

Base Sequence↗
PubMed1995

Hair cell regeneration in the chick inner ear following acoustic trauma: ultrastructural and immunohistochemical studies.

The regeneration of hair cells in the chick inner ear following acoustic trauma was examined using transmission electron microscopy. In addition, the localization of proliferation cell nuclear antigen (PCNA) and basic fibroblast growth factor (b-FGF) was demonstrated immunohistochemically. The auditory sensory epithelium of the normal chick consists of short and tall hair cells and supporting cells. Immediately after noise exposure to a 1500-Hz pure tone at a sound pressure level of 120 decibels for 48 h, all the short hair cells disappeared in the middle region of the auditory epithelium. Twelve hours to 1 day after exposure, mitotic cells, binucleate cells and PCNA-positive supporting cells were observed, and b-FGF immunoreactivity was shown in the supporting cells and glial cells near the habenula perforata. Spindle-shaped hair cells with immature stereocilia and a kinocilium appeared 3 days after exposure; these cells had synaptic connections with the newly developed nerve endings. The spindle-shaped hair cell is considered to be a transitional cell in the lineage of the supporting cell to the mature short hair cell. These results indicate that, after acoustic trauma, the supporting cells divide and differentiate into new short hair cells via spindle-shaped hair cells. Furthermore, it is suggested that b-FGF is related to the proliferation of the supporting cells and the extension of the nerve fibers.

Acoustic Stimulation↗
PubMed1995

Lysinuric protein intolerance in pregnancy: case report with successful outcome.

We present a 28-year-old pregnant woman who exhibited intolerance to lysinuric protein. However, she gave birth to a normal child without serious maternal complications. Since four of seven similar patients reported previously developed serious complications such as hemorrhage and eclampsia, patients with lysinuric protein intolerance are considered to be at high risk during pregnancy. A successful outcome can be achieved, provided that the anemia, thrombocytopenia, toxemia and hyperammonemia are well controlled.

Adult↗
PubMed1995

Antitumor effects of oral administration of an interferon-inducing pyrimidinone, Bropirimine, on murine renal-cell carcinoma.

Bropirimine [2-amino-5-bromo-6-phenyl-4-(3H)-pyrimidinone] is a low-molecular-weight compound that acts as an inducer of interferon in several animal species. Experiments were designed to explore the possibility of using this drug for the treatment of renal-cell carcinoma (RCC). Euthymic BALB/c mice were inoculated with murine RCC (Renca) cells and given graded doses of Bropirimine p.o. for 5 consecutive days beginning on day 1 following tumor inoculation. These mice were killed and tumors were excised on day 21. Bropirimine significantly (P < 0.01) inhibited the tumor growth at a daily dose of 1,000 or 2,000 mg/kg. No adverse effect or toxicity was noted at 1,000 mg/kg, and at 2,000 mg/kg there was only a marginal body-weight reduction without any other appreciable side effect. In addition to the inhibition of tumor growth, there was a small yet significant (P < 0.05) increase in the duration of survival (in days) in the Bropirimine-treated animals. When the treatment was delayed to begin on day 6 following tumor inoculation, Bropirimine did not suppress tumor growth in euthymic mice, pointing to the importance of the timing of the treatment. In athymic nude BALB/c mice lacking T-cells or T-cell function, Bropirimine also inhibited tumor growth (P < 0.01). The antitumor effect of this drug was abolished by pretreatment with anti-asialo GM1 serum, which eliminated natural killer (NK) activity in euthymic mice. In vivo treatment with Bropirimine augmented the cytotoxicity of lymphocytes isolated from the spleens or lungs of the tumor-bearing mice, which were active against Renca and YAC-1 cells in vitro. This activity was NK-cell-dependent as judged on the basis of the results of the in vitro complement-dependent cytotoxicity assay. Since Bropirimine induced interferon (IFN)-alpha/beta production, significantly (P < 0.05) elevating its serum concentration, and since this drug mimics the effects of IFN-alpha/beta, it seemed likely that the Bropirimine-induced NK cell augmentation we found was mediated by IFN-alpha/beta. These results suggest that Bropirimine, a booster of NK activity, may have potential as an adjunct to other therapeutic modalities in the treatment of human RCC.

Administration, Oral↗
PubMed1995

A long-term follow-up study in direct cerebral aneurysm surgery.

A long-term follow-up study covering between one and twenty-two years after surgery was carried out on 177 patients with direct operations for cerebral aneurysms. Rebleeding was detected in five out of 18 patients with incomplete operations; one patient within 6 months and four patients within 5 to 8 years after surgery. The rebleeding rate of incompletely operated patients in the first decade was similar to that of untreated patients with ruptured aneurysms. This suggests that wrapping, coating and incomplete clipping procedures provide some protection against rebleeding in the first 6 months but little in the period extending beyond 5 years after surgery. Showing two types of newly-formed aneurysms after complete surgery and enlarged residual aneurysms after incomplete surgery, the possibility of rebleeding from these aneurysms is discussed in the present report.

Cerebral Angiography↗
PubMed1995

Fetal nucleated cells in maternal peripheral blood after delivery in cases of fetomaternal hemorrhage.

OBJECTIVE: To determine how long fetal nucleated cells can be detected in maternal peripheral blood after delivery in cases of fetomaternal hemorrhage, using fluorescence in situ hybridization or polymerase chain reaction. METHODS: Nucleated cells with Y chromosomes were analyzed in 90 blood samples using fluorescence in situ hybridization with a Y-chromosome-specific DNA probe. The samples were obtained from 15 mothers (gravida 1), who were delivered of normal male neonates at University of Tsukuba Hospital, Japan, with fetomaternal hemorrhage 1 and 7 days, and 1, 3, 6, and 12 months after delivery. The 15 mothers were selected if fetal cells were detected above the ratio of one fetal cell per 3000 mothers' cells at 1 day after delivery. Polymerase chain reaction amplifying two different Y-specific fragments (a 102 base-pair fragment in the pHY10 vector, which was a highly repetitive sequence, and a single-copy, Y-specific 198 base-pair fragment) was also carried out for the analysis of genomic DNA from the same samples. RESULTS: Seven days and 1 month after delivery, Y-positive cells were detected in nine and four of 15 samples, respectively. No cells with the hybridization signal were detected in any of the 45 samples obtained 3 months or more after delivery. When we performed the polymerase chain reaction experiment amplifying the single-copy Y-specific fragment, none of the 45 samples after 3 months or more showed the expected band. However, from the same 45 samples, four obtained from different mothers showed positive results by amplification of the fragment in the pHY10 vector. Because multiple samples taken from these mothers before the collection of the positive samples showed negative results in the three experiments, these findings strongly suggest that the observed positive results were false-positives. CONCLUSION: Fetal nucleated cells in maternal peripheral blood become undetectable 3 months after delivery in most, if not all, cases of fetomaternal hemorrhage.

False Positive Reactions↗