[Comparative study of nuclear DNA content between fine needle aspirates and paraffin-embedded tissue samples of breast cancer: preliminary report].
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Biomedical subjects
Publications and source records attributed to T Kurihara.
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The structures of three forms of mouse myelin-associated glycoprotein mRNAs were determined from full-length cDNA clones. Two forms of mRNAs have been reported to be different by alternate inclusion of exon 2 and 12 in rat brain. One of the three forms of clones obtained here appeared to be a novel mRNA which lacked both the exon 2 and 12 portions, although others were identical splicing patterns to those of rat. Northern blot analysis using specific probes to mRNAs with or without the exon 2 portion in normal and quaking mouse confirmed that the splicing of exon 2 and 12 occurred independently.
The mouse 2',3'-cyclic-nucleotide 3'-phosphodiesterase gene was isolated from a mouse gene library. Restriction endonuclease mapping and DNA sequencing analysis revealed that this gene is about 6 kb long and is separated into three exons by two introns. The transcription initiation site was identified. The mouse cDNA of 2374 bp was obtained and used for the screening and analysis of the gene.
cDNA clones of human myelin-associated glycoprotein were isolated and analyzed. The combination of the two overlapping cDNA clones covered the full coding region and the complete amino acid sequence was deduced. In rat and mouse, expression of the two forms of mRNA is developmentally regulated; the mRNA without exon 12 portion is expressed mainly in the actively myelinating stage of development. Although the cDNA library used here was prepared from adult human brain poly(A)+ RNA, all five clones obtained corresponded to the mRNA without exon 12 portion.
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Four male patients and one female patient of a new family with Joseph disease are reported. Their disease was characterized by autosomal dominant inheritance, bulging eyes, rigidity and spasticity of the lower extremities, dystonia, and bradykinesia. Cerebrospinal fluid homovanillic acid level was markedly reduced. Levodopa improved dystonia. Magnetic resonance imaging revealed mild atrophy of the frontal lobe and the cerebellum and marked atrophy of the lenticular nucleus and the brain stem. Polysomnographic studies revealed non-rapid eye movement stage central type sleep apnea syndrome. This is the first report using magnetic resonance imaging and sleep apnea studies of Joseph disease.
A cDNA fragment of mouse 2',3'-cyclic-nucleotide 3'-phosphodiesterase was isolated and used as the probe for Northern blot analysis. The mRNA bands in normal mice became detectable at 10 days after birth, reached the maximum at the period of active myelination and then decreased gradually. At all stages of development studied, the mRNA bands in quaking mice were markedly reduced as compared with those in normal mice.
When nuclear localization sequences (termed NLS) are placed at the N terminus of cytochrome c1, a mitochondrial inner membrane protein, the resulting hybrid proteins do not assemble into mitochondria when synthesized in the yeast Saccharomyces cerevisiae. Cells lacking mitochondrial cytochrome c1, but expressing the hybrid NLS-cytochrome c1 proteins, are unable to grow on glycerol since the hybrid proteins are associated primarily with the nucleus. A similar hybrid protein with a mutant NLS is transported to and assembled into the mitochondria. To identify proteins that might be involved in recognition of nuclear localization signals, we isolated conditional-lethal mutants (npl, for nuclear protein localization) that missorted NLS-cytochrome c1 to the mitochondria, allowing growth on glycerol. The gene corresponding to one complementation group (NPL1) encodes a protein with homology to DnaJ, an Escherichia coli heat shock protein. npl1-1 is allelic to sec63, a gene that affects transit of nascent secretory proteins across the endoplasmic reticulum. Rothblatt, J. A., R. J. Deshaies, S. L. Sanders, G. Daum, and R. Schekman. 1989. J. Cell Biol. 109:2641-2652. The npl1 mutants reported here also weakly affect translocation of preprocarboxypeptidaseY across the ER membrane. A normally nuclear hybrid protein containing a NLS fused to invertase and a nucleolar protein are not localized to the nucleus in npl1/sec63 cells at the nonpermissive temperature. Thus, NPL1/SEC63 may act at a very early common step in localization of proteins to the nucleus and the ER. Alternatively, by affecting ER and nuclear envelope assembly, npl1 may indirectly alter assembly of proteins into the nucleus.
Acetoacetyl-CoA thiolase (Thiolase I) and 3-ketoacyl-CoA thiolase (Thiolase III) found in peroxisomes of an n-alkane-utilizing yeast, Candida tropicalis pK 233, were each purified to homogeneity by successive column chromatographies. Thiolase I was composed of six identical subunits whose molecular masses were 41,000 Da, and Thiolase III was a homodimer composed of 43,000 Da subunits. The results of limited proteolysis of the respective thiolases indicated that they were quite different in peptide components. Furthermore, these enzymes were immunochemically distinguishable. The kinetic studies showed that the substrates with long chains were degraded exclusively by Thiolase III, while acetoacetyl-CoA was degraded preferentially by Thiolase I. Thus, in the yeast, the complete degradation of fatty acids is suggested to be carried out efficiently in peroxisomes.
Newly synthesized pyrrolidine derivatives inhibited rabbit platelet aggregation induced by platelet activating factor.
To investigate myocardial perfusion in silent myocardial ischemia, we performed exercise stress myocardial tomography with thallium-201 (T1) in 85 patients with coronary artery disease (CAD). Exercise stress myocardial tomography was obtained both immediately after exercise and three hours later. Patients were classified into two groups according to the presence (Symptomatic Group, n = 36) or absence (Silent Group, n = 49) of chest pain during exercise stress. Clinical features (age, gender and history of myocardial infarction) and arteriographically determined severity of CAD were the same in both groups. The extent of myocardial ischemia (% Ischemia) estimated by exercise stress myocardial tomography was the same in each group (30 +/- 10% in Silent Group, 28 +/- 12% in Symptomatic Group, NS). The severity of exercise-induced myocardial ischemia was expressed as a minimal value of myocardial T1 washout rate (minimal WOR) of each patient. Although exercise heart rate was identical in both groups, minimal WOR in Silent Group was significantly higher than that of Symptomatic Group (4 +/- 10% vs -16 +/- 14%, p less than 0.001). The study in patients who exhibited both silent and symptomatic ischemia showed same results. These findings suggest that the severity of ischemia is a fundamental factor in determining the presence or absence of pain during exercise induced ischemia.
To test the diagnostic accuracy and safety of dipyridamole-loading myocardial imaging with thallium-201 (T1) in the elderly, we performed myocardial tomography in 42 patients with coronary artery disease (CAD). While monitoring of ECG and blood pressure, dipyridamole was infused at a rate of 0.57 mg/kg body weight over 4 minutes, and 4 minutes later T1 (3 mCi) was injected. Five minutes later (initial) and 3 hours after T1 injection (redistribution) myocardial images were obtained from 32 projections by a rotating gamma camera. After low-pass filtering, images were reconstructed into short-axis, horizontal long-axis and vertical long-axis tomograms. Myocardial T1 distribution both on initial and redistribution images and myocardial T1 washout rate's (WOR) of the entire left ventricle were expressed into two-dimensional polar maps by using short-axis cuts (bull's-eye maps). In addition to visual interpretation, abnormal WOR (less than 18%) was used as a criteria of dipyridamole induced myocardial ischemia. Patients were divided into 2 groups. Group I consisted of 24 patients more than 70 years of age and Group II consisted of 18 patients less than 65 years of age. The clinical and coronary angiographic characteristics of the two groups did not differ. The sensitivity of this method for the detection of CAD was identical (83%) between the 2 groups, and the detection of coronary artery stenosis was 73% in Group I and 74% in Group II (NS).(ABSTRACT TRUNCATED AT 250 WORDS)
A case of a surgically treated primary quartet cancer is reported, along with a review of the Japanese literature. A 68-year-old woman, curatively operated for a right breast cancer 6 years previously, was then diagnosed as having a left renal cell carcinoma, a thyroid cancer, and a sigmoid colon cancer as a result of a subsequent screening examination within one year. These synchronous, triple cancers were resected in a curative operation for each. Histopathologically, each neoplasm was proven to be a primary cancer lesion, and two different types (papillary and trabecular) were found in the thyroid gland.
A cDNA clone for the mRNA of bovine ARPP-21 (cAMP-regulated phosphoprotein, Mr = 21,000 as determined by SDS-PAGE) was isolated from a modified Okayama-Berg plasmid library. Transformed Escherichia coli colonies were screened by in situ colony hybridization with 2 different oligonucleotide probes derived from the amino acid sequence of the bovine protein. Sequence analysis of the longest cDNA clone, pTKAI [2407 nucleotides plus a poly(A) tail], revealed a 267-nucleotide-long coding region in agreement with the bovine ARPP-21 amino acid sequence (Williams et al., 1989). Southern blot analysis of total bovine genomic DNA raised the possibility that there may be 2 genes coding for ARPP-21. Northern blot analysis of total cellular RNA from bovine caudate nucleus and other brain regions demonstrated the existence of 2 major mRNA species, 2.5 and 1.0 kb in length, probably derived from use of alternate polyadenylation sites. There was a differential expression of these 2 mRNAs within the brain. Both ARPP-21 mRNAs were most abundant in the caudate nucleus, where the concentration of the protein is highly enriched.
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To evaluate the usefulness of preoperative exercise stress (Ex) myocardial single photon emission CT (SPECT) with thallium-201 to predict the responses to coronary revascularization (CRV), Ex-SPECT's were obtained in 42 patients with coronary artery disease (CAD). In 34 patients angioplasty was performed and in 18 patients coronary bypass surgery was undergone. Before and after CVR, Ex-SPECT's were obtained both at immediately after Ex (Initial) and 3 hours later (RD) by the rotating gamma camera. Initial images before CRV showed definite perfusion defects (+3) in 76 myocardial segments. Perfusion abnormalities at RD images were graded into 4 (+3 to 0) by visual interpretation. "+3" indicated fixed defect and "0" indicated no perfusion abnormality. At RD images 17 segments showed fixed defect and 59 segments showed improved perfusion more than one grade. After CRV, all 59 segments with improved perfusion at RD images showed improvement of perfusion in comparison with initial images before CRV. Out of 17 segments with fixed defect before CRV, 14 segments showed perfusion defect with +3, while 3 segments showed improved perfusion after CRV. These 3 segments had ECG evidence of myocardial infarction. In these 3 segments, Ex-SPECT's before CRV showed abnormally low myocardial T1 washout rate (WOR) despite they indicated fixed defect visually. On the contrary, other 14 segments with fixed defect showed normal WOR before CRV. In conclusion, visually interpreted Ex-SPECT's before CRV predict the myocardial perfusion after CRV in most of cases.2+n a small
Bulfield and others found X-linked muscular dystrophic (mdx) mouse by screening C57 BL/10 mice. The serum CK and PK are high in mdx mice, and they develop muscle degeneration 10-15 days after birth. The regeneration is vigorous in mdx mice and almost all the muscle fibers are replaced by regenerated fibers by 60 days after birth. Although mdx mice have been developed as a model for X-linked muscular dystrophy we have found that myotonic bursts are recorded when a glass microelectrode is inserted into the muscle fibers of hemidiaphragm preparations of mdx mice. Insertion myotonia is ceased by addition of the Na channel blocker tetrotoxin. Myotonia is not reduced, nor ceased by lowering the extracellular Ca to 1/15 of the volume of ordinary Tyrode's solution. Calcium antagonist, nicardipine at the dose of 10(-7), and 10(-6)M/L do not reduce myotonic bursts. Higher dose of nicardipine up to 2 x 10(-5)M/L abolished myotonic bursts. These results indicate that myotonic bursts are related to muscle membrane abnormalities, and each action potential occurs through Na channel, but not through Ca channel Higher dose of calcium antagonist can abolish myotonia by affecting Na channel in addition to their primary effects of Ca channel. The clinical effects of the Ca antagonist for myotonia was reported in one study. Since previous medications for myotonia including quinine HCl, procaine amide, diphenylhydantoin, and carbamazepine have some side effects such as tinnitus, headache, nausea, cardiac blocks, and bone marrow suppression, Ca antagonist may be used as a safe therapeutic drug for myotonia.