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Biomedical subjects

T Mita

Publications and source records attributed to T Mita.

At least 73 records · Page 4Linked to original sources

Myosin light chain kinase inhibitors ML-7 and ML-9 inhibit mouse lung carcinoma cell attachment to the fibronectin substratum.

We studied the effects of various protein kinase inhibitors on the attachment of mouse lung carcinoma 3LL cells to the fibronectin (FN) substratum. Calmodulin antagonists (W-7 and W-13) and myosin light chain kinase inhibitors (ML-7 and ML-9) exhibited the inhibitory effect for the attachment, while inhibitors of protein kinases A and C were ineffective. Since Arg-Gly-Asp-containing hexapeptide blocked the attachment, cell surface FN receptor appeared to be involved in this mechanism. These results support the hypothesis that the cell attachment requires the rearrangement of the cytoskeleton in association with the phosphorylation of myosin light chain which would lead to the clustering of the cell surface FN receptors.

Animals↗

Biochemical abnormalities in developing E1 mouse.

Complex biochemical abnormalities were found in the early developmental stage of the E1 mouse. First, the E1 mouse has abnormal levels of specific amino acid concentration within a week from birth. Second, an unusual expression of poly(A)+ RNA from the one-day newborn liver of the E1 mouse was detected by use of Cot 100 DNA as a probe. Third, sodium channels are increased in synaptosomes and at the mRNA expression level of the 3 or 4-week-old E1 mouse brains, compared with the ddY mouse. These results suggest that the biochemical abnormalities described in this study may affect greatly the epileptogenesis of E1 mouse.

Age Factors↗

Molecular cloning and expression of a novel homeobox gene AHox1 of the ascidian, Halocynthia roretzi.

We have isolated a novel ascidian homeobox gene, designated AHox1, by screening the genomic DNA of Halocynthia roretzi with the Bombyx mori Antennapedia type homeobox as a probe. The AHox1 gene encodes a protein that consists of 741 amino acids. The homeobox of AHox1 is interrupted by 2 introns each of which is about 300 bp in length and it shows about 70% similarity at a deduced amino acid level to that of Drosophila H2.0. This suggests that AHox1 is one of the most diverged homeobox genes so far characterized. Northern blot hybridization with an AHox1 probe showed the presence of single transcripts approximately 2.8 kb in length in larvae, juveniles and some adult tissues. The expression of AHox1 is scarcely detected during the course of early development but it increases to a moderate level at the larval stage. After metamorphosis, the level of AHox1 expression increases as development proceeds. In situ hybridization to the juvenile 7 days after metamorphosis showed that the site of AHox1 expression is the epithelium of digestive tract. Among the adult tissues examined, digestive tract, digestive gland and coelomic cells were the major sites of the expression of AHox1. In gonad, body wall muscle and pharyngeal epithelium, the expression of AHox1 is relatively weak. These results suggest that AHox1 is primarily expressed in the tissues of endodermal origin and that the gene expression may be associated with differentiation of the endodermal tissues.

Amino Acid Sequence↗

High resolution NMR spectroscopy using a recursive algorithm.

When the DFT (discrete Fourier transform) is used as a method for obtaining NMR (nuclear magnetic resonance) spectrum, resolution of the obtained NMR spectrum depends clearly on spin-spin relaxation times of the nucleus and the sampled number of NMR signal. That is, the NMR spectrum is affected by decay of NMR signal due to the relaxation times and errors associated with discrete approximation of NMR spectrum. As a results, its resolution is degraded and the spectrum disagrees with the density distribution of the nucleus. In the present paper, we propose a new NMR spectrum estimation method for reducing their effects, demonstrating a high resolution NMR spectrum which is closer to the density distribution than that of the DFT. The proposed method is derived from applying the Kalman filter or the recursive least-square method to a NMR signal model based on Bloch equation. Under various conditions, the proposed method is compared with the DFT using a proton 1H NMR signal observed from mayonnaise in a 2 [T] static magnetic field. The results show that if spin-spin relaxation times are known previously, the proposed method can provide a higher resolution NMR spectrum than the DFT. In addition, it is shown that the method can reduce the discrete approximation errors of NMR spectrum. Finally, we demonstrate that if frequency components of sources of NMR signal are present in a limited range in frequency domain of interest, the method can magnify NMR spectrum with a sampling time and interval fixed.

Algorithms↗

Immunohistological detection of H-2 antigens in vivo induced by IFN-beta in mouse brain tissues.

We describe the immunohistological detection of in vivo interferon-beta (IFN-beta)-induced H-2 antigens in mouse brain tissues. Two injections of recombinant mouse interferon-beta (rIFN-beta) were given into the lateral ventricles. Following perfusion with periodate-lysine-paraformaldehyde (PLP) solution, microsliced brain sections were incubated with anti H-2 monoclonal antibody and these subjected to a streptavidin biotin immunohistochemistry technique. Expression of H-2 antigens was demonstrated on neurons, neuroglias and vascular endothelial cells in the hippocampus, cerebral cortex, corpus callosum and other regions of the brain as well as on ependymocytes in the choroid plexus. H-2 antigens were also diffusely distributed in the cytoplasm and axons.

Animals↗

[3H]GBR-12935 binding sites in human striatal membranes: binding characteristics and changes in parkinsonians and schizophrenics.

The binding of the diphenyl-substituted piperazine derivative, [3H]GBR-12935, a selective dopamine uptake inhibitor, to the post-mortem human putamen was studied. Inhibition curves by dopamine uptake inhibitors suggested the existence of two populations of [3H]GBR-12935 binding sites: one is potently inhibited by mazindol and/or nomifensine, and the second binding site is benztropine- and/or GBR 12909-sensitive. In the human putamen, [3H]GBR-12935 labeled both these two distinct binding sites. The [3H]GBR-12935 binding displaced by mazindol was enriched in the mouse and rat striatum, but not in the cultured mouse neuroblastoma cell N1E-115. The mazindol-sensitive [3H]GBR-12935 binding site increased in the presence of sodium and markedly decreased in the putamen from parkinsonians (45% of controls). On the other hand, the [3H]GBR-12935 binding displaced by benztropine showed no sodium-dependent increase and was not decreased in the putamen from parkinsonians. In the putamen from schizophrenics, the [3H]GBR-12935 binding did not significantly change in the density, while that displaced by mazindol tended to increase. It is concluded that in the human putamen, [3H]GBR-12935 binds to two distinct sites. One site is partially sodium-dependent and appears to be associated with a high-affinity dopamine uptake system on dopaminergic nerve terminals. The second binding site shows no sodium-dependency and may be associated with a nondopaminergic and/or extraneuronal DA uptake system.

Aged↗

Characterization of two types of histone H2B genes from macronuclei of Tetrahymena thermophila.

Two histone H2B gene clones were isolated from macronuclei of Tetrahymena thermophila. Nucleotide sequences of the two clones were highly homologous within the coding region but not in the noncoding region. Comparison of the deduced amino acid sequences between the two clones showed three differences in a total of 121 amino acids. Each of the two clones contained a TAA triplet within the coding region, which appeared to code for a glutamine residue. To demonstrate the existence of histone mRNA containing UAA triplet, nuclease P1 protection mapping using total cellular RNA and nucleotide sequencing of primer extension products were carried out. The results clearly indicated that two cloned histone H2B genes were transcribed, giving rise to the major histone H2B mRNAs with a UAA triplet sequence in frame. The tentative 5'- and 3'-ends of histone H2B mRNAs were determined.

Amino Acid Sequence↗

Susceptibility of chimera mice to epileptic convulsion.

The ddY mouse-derived E1 strain is a mutant susceptible to severe epileptic convulsion. Using a method of aggregation of embryos from the F1 progenies of ddY female X El male and ddY female X C57BL/6J male, seven chimera mice were produced. These seven chimera mice manifested distribution of white coat color derived from F1 progeny of ddY X El, in two 100%, one 50%, two 24%, and two less than 4% and the frequency of the seizure occurrence were 71.4%, 50%, 28.6%, 28.6%, 21.4%, 0.7% and 0%, respectively. It was found that the chimera mouse which had a larger percent of white coat color had higher susceptibility to epileptic convulsion, the susceptibility having been proportional to the degree of the tissue mosaicism.

Animals↗

[3H]muscimol binding sites increased in autopsied brains of chronic schizophrenics.

[3H]muscimol binding and glutamic acid decarboxylase (GAD) activity in the prefrontal cortex and caudate nucleus of autopsied brains from 19 chronic schizophrenics and 17 control subjects were investigated. In the schizophrenics, saturation analysis with varying concentrations of [3H]muscimol revealed an increase in the number of GABAA receptors, but there was no significant difference in the affinity. In addition, the enhancement of [3H]muscimol binding by diazepam was significantly greater in schizophrenics than in controls. GAD activity did not differ between controls and schizophrenics. The possibility that GABAergic mechanisms might play a role in case of chronic schizophrenia should be given further attention.

Adult↗

Crossbreeding analysis of the mouse epilepsy.

Genetic control of the mouse epilepsy of E1 mouse isolated from ddY strain was investigated by crossbreeding with C57BL/6 mouse. A concept of the dominance deviation was applied for the explanation of the epileptic seizure inheritance in the generations of P1, F1, F2, B1 (F1 X E1 mouse) and B2 (F1 X C57BL/6 mouse), in which initial rate increase of the seizure frequency, seizure-inducing rotation numbers and seizure formative period as a function of the population growth were undertaken to be phenotypic measures. It was conclusive that the mouse epilepsy is apparently under genetic control by at least single gene with the seizure activity influencing factor(s) or by plural genes. Those are partially recessive for the allele assumed in cross-hybridized C57BL/6 mouse. No relevance of the coat color genotype and sex determination to the seizure segregation was observed obviously. Possible existence of epileptic substance(s) responsible for the enhancement of the seizure activity is also discussed.

Animals↗

The role of renal kallikrein-kinin system and prostaglandins in diuresis and natriuresis following saline infusion in normotensives and essential hypertensives.

In order to clarify the significance of the renal kallikrein(KAL)-kinin(KIN) system and prostaglandins (PG) in exaggerated natriuresis in essential hypertensives, the effect of acute sodium load on urinary KAL, KIN, PG, and renal water and sodium handling were investigated in normotensives (NT) and patients with essential hypertension (EHT). Nine NT and seven EHT were studied following acute physiological saline infusion (1000 ml/2 hrs). Urine volume (UV), urinary sodium excretion (UNaV), fractional excretion of sodium (FENa), and fractional excretion of inorganic phosphorus (FEP) were measured by the clearance method. Urinary KAL and KIN were determined by direct-RIA. Urinary kininase (total, I and II) activities were measured by the kinin destroying capacity. Urinary PGE was measured by RIA. Following saline infusion, UV, UNaV, FEP, KAL, KIN and PGE significantly increased in both NT and EHT. The increases of UV, UNaV, FENa, FEP and KAL were remarkably greater in EHT than each in NT, while no significant difference was found in the increment of PGE between NT and EHT. Significantly positive correlations were observed between PGE and KAL or KIN in NT (r = 0.889, p less than 0.005; r = 0.574, p less than 0.05, respectively), but not in EHT. From these results, it was concluded that the exaggerated natriuresis observed in EHT following infusion may be significantly related to the augmentation of renal KAL-KIN system, but was not directly related to PGE.

Diuresis↗

A comparative study of the measurement of urinary kallikrein by various methods in patients with essential hypertension and patients with proteinuria.

In order to investigate the validity of urinary kallikrein (KAL) measurement, comparative studies were performed among the values obtained by various methods of urinary KAL measurements. Daily urine samples were collected from 37 hospitalized normal subjects (NS, 21 essential hypertensives without complications (EHT) and 20 patients with renal diseases associated with proteinuria (PU). Urinary KAL excretions were determined by direct radioimmunoassay (RIA), kininogenase assay (K-genase), TAMe esterase assay (TAMe), and PPA-MCA (MCA) and PPA-NE amidase assay (NE). By the desalting procedure, urinary KAL levels showed significant changes in TAMe, MCA and NE, but not in d-RIA and K-genase in all three groups. In TAMe, MCA and NE, the recovery of added KAL in urine was significantly lower in non-desalted samples in both EHT and PU, but not in NS. Impaired recovery and correlations between d-RIA or K-genase and TAMe, MCA or NE in non-desalted samples were improved by desalting. Although good correlations were observed between d-RIA or K-genase and TAMe, MCA or NE in desalted samples, the slopes of curves were steeper in EHT and PU than in NS, suggesting that the synthetic substrate methods still have some problems in the KAL measurement in these pathological states, KAL inhibitor, aprotinin and gabezate mesilate did not suppress the esterclytic and amidolytic activities completely, but suppressed K-genase activity completely in PU urine samples, suggesting that certain kinds of non-KAL esterases might remain in PU urine samples. Thus, d-RIA and K-genase appear to be the most reliable methods in the measurement of urinary KAL quantity and activity, respectively.

Humans↗

Decreased serotonin S2 and increased dopamine D2 receptors in chronic schizophrenics.

Serotonin S2 and dopamine D2 receptors in the prefrontal cortex and caudate nucleus of postmortem brains of chronic schizophrenics were studied using 3H-ketanserin and 3H-spiperone, respectively. In the prefrontal cortex of schizophrenics, we found a significant decrease in the maximum number of 3H-ketanserin binding sites (Bmax), with no change in the dissociation constant (Kd). Conversely, both Bmax and Kd of 3H-spiperone binding to the caudate nucleus were significantly increased in the schizophrenic patients. There were no differences in receptor indices between patients who were taking neuroleptics until their death and those who had taken none for 2 months or more prior to death. These findings suggest that alterations in S2 receptors in the prefrontal cortex may reflect the disease process, per se, and that the increase in the number of D2 receptors in the caudate nucleus of schizophrenics is not due solely to neuroleptic medication.

Adult↗

Faithful initiation of the in vitro transcription of a cloned rDNA from Tetrahymena pyriformis.

A cloned rDNA fragment containing the transcriptional start point of Tetrahymena pyriformis was transcribed in vitro with a crude extract from homologous Tetrahymena cells. When a KpnI-HindIII fragment from the cloned rDNA plasmid was used as the template, the runoff transcription gave rise to two major RNA products about 490 and 460 nucleotides (nt) in length. An RNA of about 490 nt long was found to correspond to the expected transcript starting from the in vivo start point determined at our laboratory [Saiga et al., Nucl. Acids Res. 10 (1982) 4223-4235]. Precise size determination was performed using an RNA size marker prepared by hybridizing the template DNA with in vitro-capped 35S pre-rRNA followed by treatment with single-strand specific P1 nuclease. The size of the runoff transcript changed as predicted, according to downstream truncation points. The effects of alpha-amanitin and actinomycin D showed the transcription to be dependent on the added template DNA and to be catalyzed by form-I RNA polymerase. Possible reasons for the discrepancy between our determination of the size of the transcription product and that of Sutiphong et al. [Biochemistry 23 (1984) 6319-6326] are discussed.

Amanitins↗

Adenovirus 2 peptide IX gene is expressed only on replicated DNA molecules.

Transcription of the adenovirus type 2 peptide IX (pIX) gene was examined in transient expression assays. When a nonreplicating plasmid DNA containing the pIX gene was introduced into HeLa cells by the DEAE-dextran method, no pIX gene transcript was detected. In contrast, efficient transcription was observed in the cells transfected with a replicating plasmid containing the pIX gene. Adenovirus early genes did not affect the level of transcription of the pIX gene on either a nonreplicating or a replicating plasmid. Inhibition of plasmid replication with cytosine arabinoside prevented transcription of the pIX gene. By quantitative analysis of the amount of the pIX gene and its transcript in transfected cells, it was concluded that active transcription of the pIX gene occurred only on DNA molecules replicated in the cell.

Adenoviruses, Human↗