Observation of nuclear resonant scattering accompanied by phonon excitation using synchrotron radiation.
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Journal Article↗
Biomedical subjects
M Seto
Publications and source records attributed to M Seto.
Phonon density of states measured by inelastic nuclear resonant scattering.
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Journal Article↗
Selective inhibition of myosin phosphorylation and tension of hyperplastic arteries by the kinase inhibitor HA1077.
To examine possible alterations in myosin light chain phosphorylation in hyperplastic arteries, rabbit strips from right hyperplastic and left normal control carotid arteries were used for experiments 6 weeks after the ballooning procedure. When the hyperplastic artery was stimulated with various concentrations of K+ (10, 20, 30, 40 and 60 mM), the maximal tension in response to each concentration was significantly higher (P < 0.05) than that in the control artery. The maximal extent of myosin light chain phosphorylation induced by 60 mM K+ in the hyperplastic artery was also significantly higher than that in the control (55.1 +/- 4.1 vs. 45.1 +/- 3.2%, mean +/- S.D.). However, the [Ca2+]i response to elevated K+ in hyperplastic arteries was much the same as that in control arteries, when measured with fura-PE3. HA1077 (1-5-(isoquinolinesulfonyl)-homopiperazine), a protein kinase inhibitor, was about 3-5 times more effective in inhibiting the tension and myosin light chain phosphorylation induced by 60 mM K+ in the hyperplastic artery than in the control artery. Nifedipine inhibited the tension and myosin light chain phosphorylation to the same extent in control and hyperplastic arteries. Thus, an alteration of the myosin light chain phosphorylation system, but not an alteration of Ca2+ mobilization, may be involved in the enhanced contraction of the hyperplastic artery. The enhanced phosphorylation of myosin light chain may be sensitive to HA1077.
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
Molecular cloning of the gene coding for the mouse T-cell antigen CD7.
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Amino Acid Sequence↗
Experimental neonatal autoimmune myasthenia gravis: an immunohistochemical, ultrastructural and electrophysiological study of the motor end-plate.
Neonatal rats born of and nursed by mothers immunized with Narke japonica acetylcholine receptor protein had elevated serum anti-acetylcholine receptor antibodies that reached the mother's level on day 10 after delivery and decreased rapidly after weaning. IgG was present at the motor end-plates up to day 170, and the motor end-plate fine structure remained abnormal up to day 80. Miniature end-plate potential amplitudes in the diaphragm were at the control levels within 10 days of birth, but were lower than those of the controls up to day 80 after birth. We could not obtain the direct evidence that transient synthesis of antibodies occurs in experimental autoimmune myasthenia gravis pups. This model can serve as an experimental model of transient neonatal myasthenia gravis in humans, exception for the route of antibody transfer and the time of the onset of illness.
Animals↗
Molecular features of the CAG repeats and clinical manifestation of Machado-Joseph disease.
Machado--Joseph disease (MJD) is an autosomal dominant spinocerebellar degeneration mapped to chromosome 14q32.1. The CAG expansions of the MJD1 gene was identified as the cause of the disease. We have analyzed 90 MJD individuals from 62 independent MJD families and found that the MJD1 repeat length is inversely correlated with the age of onset (r = -0.87). The MJD chromosomes contained 61-84 repeat units, whereas normal chromosomes displayed 14-34 repeats. In the normal chromosomes, 14 repeat units were the most common and the shortest. In association with the clinical anticipation of the disease, a parent--child analysis showed the unidirectional expansion of CAG repeats and no case of diminution in the affected family. The differences in CAG repeat length between parent and child and between siblings are greater in paternal transmission than in maternal transmission. Detailed analysis revealed that a large degree of expansion was associated with a shorter length of MJD1 gene in paternal transmission. On the other hand, the increments of increase were similar for shorter and longer expansion in maternal transmission. Among the three clinical subtypes, type I of MJD, with dystonia, showed a larger degree of expansion in CAG repeats of the gene and younger ages of onset than the other types.
Adult↗
The positive nuclear staining observed with monoclonal antibody against PRAD1/cyclin D1 correlates with mRNA expression in mantle cell lymphoma.
Recently, we produced a monoclonal antibody, 5D4, against the PRAD1/cyclin D1 product and suggested positive nuclear staining to be associated with mantle cell lymphoma (MCL). Now we have further characterized the specificity of this antibody and studied the relation of immunohistochemical detection to PRAD1/cyclin D1 mRNA expression and DNA rearrangement. Immunofluorescence and immunoblotting studies demonstrated the 5D4 antibody to be crossreactive with cyclin D2, but not cyclin D3. On immunostaining, 15 of 19 MCL cases (79%) presented the nuclear staining pattern and PRAD1/cyclin D1 mRNA expression was detected by Northern blot analysis in 12 of 15 MCL cases studied (80%): all cases with the mRNA expression showed the nuclear staining pattern. Southern blot analysis with 11q13 BCL-1 probes detected DNA rearrangements in 8 of 19 MCL cases (42%), all 8 exhibiting PRAD1/cyclin D1 mRNA expression. In 21 lymphoma cases of types other than MCL, neither the mRNA expression nor the nuclear staining were observed, although cytoplasmic staining was often apparent. These results indicated that positive nuclear staining of lymphoma cells by 5D4 antibody reflects PRAD1/cyclin D1 mRNA expression, and showed that this monoclonal antibody has diagnostic value for differentiating MCL from other types of lymphomas.
Animals↗
Multiple Polychlorinated Biphenyl Transformation Systems in the Gram-Positive Bacterium Rhodococcus sp. Strain RHA1.
The cloned bphA gene of the polychlorinated biphenyl (PCB) degrader Rhodococcus sp. strain RHA1 was expressed in Rhodococcus erythropolis IAM1399 cells, resulting in the transformation of di-, tri-, and tetrachlorobiphenyls. Disruption of the bphA1 gene in RHA1 resulted in a lack of growth on biphenyl and a loss of PCB transformation activity. However, the bphA1 insertion mutant of RHA1, designated RDA1, retained the ability to transform PCB congeners when grown on ethylbenzene as its carbon source. It also transformed 4-chlorobiphenyl to 4-chlorobenzoate, although it was suspected to be deficient in bphB and bphC gene activities as well as bphA. This suggested that an alternative PCB degradation system distinct from the one encoded by the cloned bph genes was present.
Journal Article↗
A Novel Transformation of Polychlorinated Biphenyls by Rhodococcus sp. Strain RHA1.
We have characterized a biphenyl degrader, Rhodococcus sp. strain RHA1. Biphenyl-grown cells of strain RHA1 efficiently transformed 45 components in the 62 major peaks of a polychlorinated biphenyl (PCB) mixture of Kanechlors 200, 300, 400, and 500 within 3 days, which includes mono- to octachlorobiphenyls. Among the intermediate metabolites of PCB transformation, di- and trichlorobenzoic acids were identified. The gradual decrease of these chlorobenzoic acids during incubation indicated that these chlorobenzoic acids would also be degraded by this strain. The effect of the position of chlorine substitution was determined by using PCB mixtures that have chlorine substitutions mainly at either the ortho or the meta position. This strain transformed both types of congeners, and strong PCB transformation activity of RHA1 was indicated. RHA1 accumulated 4-chlorobenzoic acid temporally during the transformation of 4-chlorobiphenyl. The release of most chloride in the course of 2,2(prm1)-dichlorobiphenyl degradation was observed. These results suggested that RHA1 would break down at least some PCB congeners into smaller molecules to a considerable extent.
Journal Article↗
The time course of myosin light-chain phosphorylation in blood-induced vasospasm.
The phosphorylation of an M(r) 20,000 myosin light chain (MLC20) promotes the generation of contractile force through actin-myosin adenosine triphosphatase in most agonist-mediated vascular smooth muscle cell contraction. However, the role of calcium-mediated contractile processes in sustained arterial narrowing after subarachnoid hemorrhage remains unknown. In a femoral artery model of vasospasm, whole blood was applied to arteries in 54 rats for periods of 2 to 10 days; the contralateral artery treated with platelet-rich plasma served as matched control. During the early stage of vasospasm (Days 2-5), in the media of arteries exposed to blood, MLC20 phosphorylation (including diphosphorylated forms) increased significantly (30-38%; P < 0.05); total medial MLC20 during this interval was comparable to that in controls. After 5 days, however, total MLC20 decreased markedly (> 90%; P < 0.01) compared with controls; phosphorylated MLC20 was undetectable during this interval. MLC20-mediated contractile processes may be prominent in the early stages of arterial narrowing after subarachnoid hemorrhage; later stages are associated with the loss of MLC20 and the possible persistence of arterial narrowing by other mechanisms.
Animals↗
Thermal conditions in the bathroom in winter and summer, and physiological responses of the elderly during bathing.
Thermal conditions in the bathroom and physiological responses were examined during winter and summer. The subjects were 22 male and 20 female elderly people, between 65 and 88 years old living in 25 houses in Gunma Prefecture, Japan. Heart rate, blood pressure, skin temperature and thermal sensation were measured during bathing. Changes in thermal sensation due to bathing were assessed in the living room and dressing room on a 9-point scale. Then they were asked about the purposes of bathing and the facilities of bathroom and dressing room. The results are summarized as follows: 1. The purpose of bathing in winter was to warm up for more than 80% of the subjects. In summer, all subjects felt refreshed by bathing. Eighty-five percent of the subjects took a bath every other day in both seasons. 2. Fifty-two percent of the bathrooms had no ventilating fans and 32% had no exclusive dressing rooms. 3. The average room temperature in the dressing rooms was 13-14 degrees C in winter. Thermal sensation was 'cool', 'slightly cold' or 'cold' for more than two-thirds of the subjects when they were partially nude, and there were no heaters in most dressing rooms. 4. The heart rate increased steadily, and reached a maximum value in a partially dressed condition in both seasons. 5. In winter, a marked increase of systolic blood pressure was observed in the partially nude condition. There was a significant difference between the before bathing condition and partially nude condition in winter.(ABSTRACT TRUNCATED AT 250 WORDS)
Aged↗
Phosphorylation of calponin by PKC is blocked by F-actin in vitro.
Polyclonal antibody against phosphorylated calponin was raised in rabbits by application of the peptide corresponding to residues 183-195 of calponin phosphorylated by protein kinase C. When calponin was incubated with protein kinase C, only free calponin was recognized by this antibody and calponin of native thin filament or that binding to F-actin did not. In experiments done using [gamma-32P] ATP, no radioactivity was detected except for free calponin. Calponin phosphorylation was suppressed in an actin dose-dependent manner and the phosphorylation of calponin was completely blocked when the actin molar ratio to calponin exceeded 10. These data suggest that phosphorylation of calponin by protein kinase C was apparently blocked by F-actin.
Actins↗
A variant chromosome translocation at 11q13 identifying PRAD1/cyclin D1 as the BCL-1 gene.
The 11q13 breakpoint region of t(11;14) (q13;q32), translocated to the Ig heavy chain locus at 14q32, has been designated as BCL-1 for B-cell leukemia/lymphoma-1, but the nature of the transcriptional unit has long remained unclear. Recently, the PRAD1 gene encoding cyclin D1, isolated from the 11q13 region, was proposed as a candidate BCL-1 gene on the basis of chromosome walking and concordant overexpression of PRAD1 mRNA in cell lines with t(11;14)(q13;q32). We report here molecular analysis of a variant translocation at the BCL-1 locus, t(11;22)(q13;q11), showing juxtaposition of the Ig light chain gene, Ig lambda, to the PRAD1 gene at its 3' end, resulting in overexpression of PRAD1 mRNA. Because only the PRAD1 gene is present between the Ig heavy chain and light chain gene breakpoints, an identity between BCL-1 and the PRAD1/cyclin D1 gene is strongly indicated.
Base Sequence↗
A reverse transcriptase-polymerase chain reaction detects heterogeneous chimeric mRNAs in leukemias with 11q23 abnormalities.
The MLL gene involved in 11q23 translocations found in the majority of infantile leukemias and some secondary leukemias makes fusion transcripts with genes such as LTG4 (chromosome 4), LTG9 (chromosome 9), and LTG19 (chromosome 19) as a result of reciprocal translocation. We have examined 25 cases of leukemias with 11q23 abnormalities by Southern blot analysis and the reverse transcriptase-polymerase chain reaction (RT-PCR). Using various primer pairs, chimeric mRNAs could be amplified in 6 of 7 leukemias with t(4;11), 6 of 8 leukemias with t(9;11) including secondary leukemia, 8 of 9 leukemias with t(11;19), and 1 with a deletion at 11q23. The chimeric mRNAs were heterogeneous and differential usage of the MLL exons was found, irrespective of the partner chromosomes. Sensitivity studies showed that a single clone with chimeric mRNA in 10(4) to 10(5) cells could be detected. These findings show that the present RT-PCR settings provide a rapid, accurate, and sensitive tool for diagnosing leukemias with 11q23 translocations and for monitoring response to therapy in these patients.
Adult↗
Messenger RNA levels of five genes located at chromosome 11q13 in B-cell tumors with chromosome translocation t(11;14)(q13;q32).
To identify genes activated by chromosome translocation t(11;14)(q13;q32), mRNA levels of five genes (cyclin D1, EXP1, MB38, HST1, and INT2) at chromosome 11q13 were investigated. The cyclin D1 mRNA increased in BCL-1-rearranged B-cell tumor cell lines SP-49, NOP-2, FLAM-76, KMS-12-PE, and KMS-12-BM cells, while it was not detected in cell lines without the translocation, Raji, U266, and HEL cells. A significant amount of the MB38 mRNA was detected irrelevantly to the translocation in all of these cell lines. The mRNAs of EXP1, HST1, and INT2 were undetectable in these cells. The results suggested that the translocation activates cyclin D1 alone, while the mRNA levels of the other four genes are regulated independently of the translocation.
Blotting, Northern↗
Prevention of TCR-mediated apoptosis by the elevation of cAMP.
The stimulation through TCR-CD3 complexes by immobilized anti-CD3 antibody induced the production of IL-2 and activation-induced cell death (ACD) in the majority of T cell hybridomas. However, some hybridomas produced IL-2 without showing any signs of ACD by the same stimulation, indicating that TCR-CD3-mediated signaling pathways of IL-2 production and of ACD are different. These pathways were discriminated from each other by protein kinase inhibitors and cAMP-elevating reagents such as forskolin. The pathway of IL-2 production but not of ACD was inhibited by protein kinase inhibitors. On the other hand, various cAMP-elevating reagents prevented the T cell hybridomas from TCR-mediated ACD with minimal inhibition of IL-2 production. The elevated cytoplasmic cAMP did not block dexamethasone-induced apoptosis. This indicates that apoptosis is regulated by multiple pathways. Furthermore, the inhibitory effect of cAMP is specific for the TCR-mediated signaling pathway of ACD. Messenger RNA for bcl-2 was detected after treatment with forskolin.
Animals↗
An antibody for phosphorylated myosin light chain of smooth muscle: application to a biochemical study.
An antibody with specificity for the 20 kDa myosin light chain of smooth muscle phosphorylated by myosin light chain kinase was developed. The antibody was raised against the phosphorylated synthetic peptide, Lys-Lys-Arg-Pro-Gln-Arg-Ala-Thr-phospho-Ser-Asn-Val-Phe-Cys (residues 11-22 of the myosin light chain). This antibody only recognized the monophosphorylated myosin light chain at serine 19, i.e., with no detectable recognition of nonphosphorylated or diphosphorylated serine 19 or threonine 18. The specificity was tested by EIA assaying of myosin light chain kinase activity using a 96-well plate coated with the light chain. This EIA system was as sensitive as the radioisotopic method, and the newly developed non-radioisotopic method.
Amino Acid Sequence↗
Loss of heterozygosity at the bcl-2 gene locus and expression of bcl-2 in human gastric and colorectal carcinomas.
The loss of heterozygosity (LOH) at the bcl-2 gene locus and the expression of the bcl-2 gene were examined in gastric and colorectal carcinoma cell lines and carcinoma tissues. LOH at the bcl-2 locus was detected in 24% (4/17) of gastric and 60% (6/10) of colonic carcinomas, all of which were well differentiated adenocarcinomas, whereas LOH was not seen in poorly differentiated ones. On the other hand, 24% (5/21) of poorly differentiated stomach cancers overexpressed bcl-2 gene, whereas no overexpression was detected in well differentiated stomach cancer. Three gastric and three colorectal carcinoma cell lines, all of which were derived from poorly differentiated adenocarcinomas, expressed considerable levels of bcl-2 mRNA and protein. These results suggest that LOH at the bcl-2 locus is frequently associated with well differentiated adenocarcinomas of the stomach and colon, and bcl-2 overexpression has implications for the development of poorly differentiated adenocarcinomas of the gastrointestinal tract.
Adenocarcinoma↗