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T Haraguchi

Publications and source records attributed to T Haraguchi.

At least 55 records · Page 3Linked to original sources
PubMed1994

Telomere-led premeiotic chromosome movement in fission yeast.

The movement of chromosomes that precedes meiosis was observed in living cells of fission yeast by fluorescence microscopy. Further analysis by in situ hybridization revealed that the telomeres remain clustered at the leading end of premeiotic chromosome movement, unlike mitotic chromosome movement in which the centromere leads. Once meiotic chromosome segregation starts, however, centromeres resume the leading position in chromosome movement, as they do in mitosis. Although the movement of the telomere first has not been observed before, the clustering of telomeres is reminiscent of the bouquet structure of meiotic-prophase chromosomes observed in higher eukaryotes, which suggests that telomeres perform specific functions required for premeiotic chromosomal events generally in eukaryotes.

Cell Nucleus↗
PubMed1993

Glucocorticoids coordinately disrupt a transforming growth factor alpha autocrine loop and suppress the growth of 13762NF-derived Con8 rat mammary adenocarcinoma cells.

We have demonstrated previously that the synthetic glucocorticoid dexamethasone suppresses the growth of Con8 rat mammary tumor cells, which are derived from the 13762NF transplantable, hormone-responsive rat mammary adenocarcinoma. Dexamethasone inhibited [3H]thymidine incorporation into Con8 cells at high cell density under both serum and serum-free conditions. Fractionation in nonreducing sodium dodecyl sulfate-polyacrylamide gels of proteins secreted from dexamethasone-treated and untreated Con8 mammary tumor cells revealed two size classes of glucocorticoid inhibited mitogenic activities; a larger M(r) 27,000-33,000 and a smaller M(r) 5,000-12,000 activity. Both size classes of mitogens restimulated the growth of glucocorticoid-suppressed Con8 cells suggesting that they can act in an autocrine fashion. The smaller mitogen was identified as transforming growth factor alpha (TGF-alpha) since this activity competed with 125I-epidermal growth factor (EGF) for EGF receptor binding and was selectively immunodepleted with monoclonal TGF-alpha antibodies but not with EGF antibodies. Western blots and radioreceptor assay of Con8-secreted proteins revealed that glucocorticoids inhibited the production of a M(r) 5500 immunoreactive TGF-alpha protein by 10-fold. Consistent with a steroid effect on the level of TGF-alpha production, rather than on its activity, the specific mitogenic activities of the TGF-alpha s secreted by dexamethasone-treated and untreated Con8 cells were identical to that of recombinant human TGF-alpha. Treatment of intact cells with suramin, which dissociates ligand-receptor complexes, revealed that the EGF receptor-mediated mitogenic response is functional in both glucocorticoid-treated and untreated cells. Taken together, our results demonstrate that glucocorticoids suppress Con8 mammary tumor cell growth and disrupt a potential TGF-alpha autocrine loop which results in a dramatic reduction in the level of extracellular TGF-alpha.

3T3 Cells↗
PubMed1992

[Functional pulmonary atresia: a case report].

A male neonate, two days of age, was admitted with cyanosis and tachypnea. Chest roentgenograms showed massive cardiomegaly and decreased pulmonary vasculatures. Echocardiographic and the right ventriculographic findings were interpreted as showing primary pulmonary atresia with secondary tricuspid incompetence. At a Brock operation, however, there was no resistance in passing the instrument through the pulmonary valve. The baby died twelve hours after surgery. Postmortem examination showed that the tricuspid regurgitation was a cause of the functional pulmonary atresia (FPA). It is important to distinguish FPA from pure pulmonary atresia because conservative therapy is the first choice for the FPA.

Diagnosis, Differential↗
PubMed1991

Identification of the glucocorticoid suppressible mitogen from rat hepatoma cells as an angiogenic platelet-derived growth factor A-chain homodimer.

We have previously shown that BDS.1 rat hepatoma cells are hypersensitive to the antiproliferative effects of glucocorticoids, and secrete a glucocorticoid suppressible mitogenic activity (denoted GSM). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that GSM purified to near homogeneity migrated as a 28-kDa protein under nonreducing conditions and as a single 15-kDa polypeptide in the presence of sulfhydryl reducing agents suggesting a homodimeric structure. Anti-platelet-derived growth factor (PDGF) A-chain specific antibodies selectively immunodepleted the mitogenic activity which can be extracted from nonreducing gels in the 26-30-kDa fraction and, in Western blots, recognized the 15-kDa reduced form of GSM. Western blot analysis further showed that dexamethasone suppressed the level of secreted PDGF A-chain protein in BDS.1 cells but not in glucocorticoid receptor-deficient hepatoma cells. Northern blots revealed that dexamethasone reduced expression of the PDGF A-chain 2.3- and 1.7-kilobase transcripts in proportion to the level of detectable PDGF-AA protein. Similarly to PDGF-AA, the hepatoma cell-derived GSM has a potent angiogenic activity. Taken together, our results demonstrate that the predominant glucocorticoid suppressible mitogen secreted from rat hepatoma cells is a PDGF A-chain homodimer and suggest that in vivo glucocorticoids may potentially regulate hepatoma growth by modulating PDGF-stimulated tumor vascularization.

Animals↗
PubMed1991

Doppler echocardiographic evaluation of an infant with HCM during ACTH therapy.

Adrenocorticotropic hormone (ACTH) therapy is useful in the treatment of patients with West syndrome, and hypertrophic cardiomyopathy (HCM) in association with ACTH therapy has recently been reported. We describe the Doppler echocardiographic evaluation of an infant who had West syndrome and HCM. ACTH therapy produced increases in the interventricular septal thickness, the pressure gradient of the left ventricular outflow tract, and the heart rate. Doppler echocardiographic examination was found to be much more sensitive for monitoring the cardiac changes than either 2-dimensional echocardiography or the heart rate.

Adrenocorticotropic Hormone↗
PubMed1990

Molecular cloning of the protocatechuate 4,5-dioxygenase genes of Pseudomonas paucimobilis.

We determined the nucleotide sequence of a 1.9-kilobase fragment of Pseudomonas paucimobilis SYK6 chromosomal DNA that included genes encoding protocatechuate 4,5-dioxygenase, the enzyme responsible for the aromatic ring fission of protocatechuate. Two open reading frames of 417 and 906 base pairs were found that had no homology with previously reported sequences, including those encoding protocatechuate 3,4-dioxygenase. Since both open reading frames were indispensable for the enzyme activity, they should encode the subunits of protocatechuate 4,5-dioxygenase. We named these genes ligA and ligB. Protocatechuate 4,5-dioxygenase was efficiently expressed in Escherichia coli with the aid of the lac promoter, and the polypeptides of the ligA and ligB gene products were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and amino acid sequencing.

Amino Acid Sequence↗
PubMed1990

[A case of non-Cushingoid Cushing's syndrome].

A 42-year-old female was admitted to our hospital with a chief complaint of hypertension. Endocrinologically, the plasma cortisol level was increased, but its diurnal rhythm had disappeared and the plasma cortisol level was not suppressed by dexamethasone loading at 2 mg or 8 mg. The plasma ACTH level was low. Computerized tomographic scan, echography and adrenocortical scintigraphy showed an adrenocortical tumor on the right adrenal gland. Physical examination did not reveal typical Cushingoid symptoms such as moon face, central obesity and violaceous striae. Thus we diagnosed this case as non-Cushingoid Cushing's syndrome and performed right adrenalectomy. Histological examination showed adreno-cortical adenoma without malignancy.

Adenoma↗
PubMed1989

Partial characterization of a glucocorticoid suppressible mitogenic activity secreted from a rat hepatoma cell line hypersensitive to the antiproliferative effects of glucocorticoids.

We have previously shown that glucocorticoids suppress the proliferation of Fu5 hepatoma cells and have selected subclones which are either hypersensitive (BDS1) or resistant (EDR3) to the antiproliferative effects of dexamethasone, a synthetic glucocorticoid. BDS1 cells externalize a glucocorticoid suppressible mitogenic activity (denoted GSM) which stimulated [3H]thymidine incorporation in quiescent, serum-starved Balb/c 3T3 cells. Glucocorticoid treatment of BDS1 cells reduced the secreted levels of GSM activity by approximately 20-fold in comparison to untreated cells. The GSM activity was constitutively secreted from a glucocorticoid receptor minus variant (EDR3) demonstrating that the suppression of this mitogenic activity is a new glucocorticoid hormone response which required a functional receptor. GSM activity was sensitive to sulfhydryl reducing agents or trypsin, stable to heat and acid treatments and fractionated in gel filtration columns with a native molecular weight of approximately Mr 30,000. The persistence of this size for mitogenic activity after electrophoretic fractionation in nonreducing sodium dodecyl sulfate-poly-acrylamide gels suggested that the GSM activity is comprised of a single protein. Total secreted protein isolated from untreated BDS1, but not dexamethasone-treated BDS1, stimulated 3T3 cells to grow in transformed-appearing large colonies in soft agar and to display multiple layering and elongated spindle-like morphology on solid substratum. The addition of both insulin and EGF to conditioned medium protein isolated from glucocorticoid-treated BDS1 cells restored full induction of 3T3 cell anchorage-independent growth while insulin restored full and EGF partial mitogenic stimulation of these fibroblasts. These results suggest that the GSM activity acts in a pathway common to that of insulin or EGF in fibroblasts.

Animals↗
PubMed1989

Detection and localization of a new enzyme catalyzing the beta-aryl ether cleavage in the soil bacterium (Pseudomonas paucimobilis SYK-6).

Cleavage of the arylglycerol-beta-aryl ether linkage is the most important process in the biological degradation of lignin. We determined the activity of the enzyme cleaving the beta-aryl ether linkage in membranes of Pseudomonas paucimobilis SYK-6. This enzyme was tightly associated with the cellular membrane and catalyzed the unique and reductive cleavage of compound II but not cleavage of compound I. This enzymatic activity was stimulated by addition of NADH. On the basis of this evidence, we present a model of the specific cellular assimilation of beta-aryl ether by P. paucimobilis SYK-6.

Bacterial Proteins↗
PubMed1989

Asparagine-linked glycosylation of the scrapie and cellular prion proteins.

Post-translational modification of the scrapie prion protein (PrP) is thought to account for the unusual features of this protein. Molecular cloning of a PrP cDNA identified two potential Asn-linked glycosylation sites. Both the scrapie (PrPSc) and cellular (PrPC) isoforms were susceptible to digestion by peptide N-glycosidase F (PNGase F) but resistant to endoglycosidase H as measured by migration in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PNGase F digestion of PrPC yielded two proteins of Mr26K and 28K; however, the 26-k species was only a minor component. In contrast, PNGase F digestion of PrPSc yielded equimolar amounts of two proteins of Mr26K and 28K. The significance of this altered stoichiometry between the 26- and 28-kDa deglycosylated forms of PrP during scrapie infection remains to be established. Both isoforms as well as PrP 27-30, which is produced by limited proteolysis of PrPSc, exhibited a reduced number of charge isomers after PNGase F digestion. The molecular weight of PrP 27-30 was reduced from 27K-30K by PNGase F digestion to 20K-22K while anhydrous hydrogen fluoride or trifluoromethanesulfonic acid treatment reduced the molecular weight to 19K-21K and 20K-22K, respectively. Denatured PrP 27-30 was radioiodinated and then assessed for its binding to lectin columns. PrP 27-30 was bound to wheat germ agglutinin (WGA) or lentil lectins and eluted with N-acetylglucosamine or alpha-methyl-mannoside, respectively. Digestion of PrP 27-30 with sialidase prevented its binding to WGA but enhanced its binding to Ricinus communis lectin. These findings argue that PrP 27-30 probably possesses Asn-linked, complex oligosaccharides with terminal sialic acids, penultimate galactoses, and fucose residues attached to the innermost N-acetyl-glucosamine. Whether differences in Asn-linked oligosaccharide structure between PrPC and PrPSc exist and are responsible for the distinct properties displayed by these two isoforms remain to be established.

Animals↗
PubMed1989

A simplified Master's two-step test for preschool children.

To investigate the usefulness of a simplified Master's two step test (s-MTT) for preschool children aged 4-6, s-MTT was carried out in our pediatric cardiology clinic using a new stair and connector for joining the leads from each child to the ECG machine. The subjects were 21 children with Kawasaki disease without coronary involvement and four children with arrhythmias. The treadmill exercise test (TET) was done on the same day to compare the exercise load of s-MTT with that of TET. Oxygen uptake during s-MTT was also examined using another group consisting of eight outpatients. There were no differences in indices such as peak heart rate (HR), HR immediately after exercise, and performance ratio between the outpatients in this study and kindergarteners in the preliminary report. The proportions of peak HR and HR immediately after exercise in s-MTT to those in TET were 91.1 +/- 7.9 (mean +/- SD)% and 91.1 +/- 8.6%, respectively. The average oxygen uptake of the eight outpatients in s-MTT was 23.2 +/- 4.1 ml/min/kg. These studies suggest that the simplified MTT is useful as a screening test, because it can be done for all children aged 4-6 in any laboratory.

Cardiovascular Diseases↗
PubMed1988

Experimental studies on the initial focus of invasion of group A streptococci.

Rabbits were infected with group A streptococci type 12 by an aerosol method, that seems to resemble closely the human natural infection, in order to investigate the mechanism of invasion, especially the initial focus of invasion of streptococci. Living streptococci and FITC-labelled streptococci were exposed to several groups of rabbits, which were sacrificed and divided according to the duration after exposure. The specimens of organs were examined by light microscopy, fluorescent microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Histologically, the adherence of streptocossi was found on the pharyngeal mucosa and tonsils and the mucocilial clearance in the larynx and the trachea. Phagocytosis was also found in the bronchus and lung. By SEM, invading streptococci and cell debris-like masses were observed in the micropore of the tonsillar crypt of the rabbit. From these results it was supposed that there should be differences of tissue tropisms and defence mechanisms against streptococci among the mucosa and that micropores of the tonsillar crypts should play an important role in the initial focus of invasion of group A streptococci.

Aerosols↗
PubMed1987

Preferential loss of DNA polymerase alpha following suppression of replicative DNA synthesis of guinea pig macrophages by the immunostimulants muramyl dipeptide or lipopolysaccharide.

Oil-induced guinea pig peritoneal exudate macrophages were found to incorporate 3H-thymidine into trichloroacetic acid-insoluble fraction. In pulse-labeling experiments, the incorporated 3H-thymidine was detected in short fragments of DNA, which corresponded to the Okazaki fragments. These results indicate that the observed thymidine incorporation is due to nuclear DNA replication but not DNA repair. The observed DNA synthesis of the macrophages was remarkably suppressed when the cells were cultured in a presence of muramyl dipeptide (MDP) or bacterial lipopolysaccharide (LPS). The significant decrease of DNA polymerase alpha activity was found in the cells treated with MDP or LPS. In contrast, the activity of polymerase beta was not at all affected by the same treatment.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed1983

Specific inhibitors of eukaryotic DNA synthesis and DNA polymerase alpha, 3-deoxyaphidicolin and aphidicolin-17-monoacetate.

Of several phytotoxins isolated from culture filtrates of Phoma betae Frank PS-13, an incitant of leaf spot disease of sugar beet, three have been identified as aphidicolin, 3-deoxyaphidicolin and aphidicolin-17-monoacetate. Aphidicolin is a selective inhibitor of eukaryotic DNA polymerase alpha (Ikegami et al. (1978) Nature 275, 458-460). Consequently, we studied the action mechanism of 3-deoxyaphidicolin and aphidicolin-17-monoacetate. These aphidicolin analogues markedly inhibited the in vivo DNA synthesis of sea urchin embryos and HeLa cells but not RNA and protein syntheses. Only DNA polymerase alpha, not DNA polymerase beta and gamma, was inhibited by these drugs. The mode of action of these analogues on DNA polymerase alpha from the sea urchin was competitive inhibition with respect to dCTP with Ki values of 0.44 micrograms/ml for deoxyaphidicolin and 0.89 micrograms/ml for aphidicolin monoacetate, respectively. None of the other three dNTPs competed with these drugs. A similar inhibitory mode was observed using the enzyme from HeLa cells and toad oocytes. These drugs at a concentration of 2 micrograms/ml caused a delay in the cleavage of fertilized eggs of the sea urchin and decomposition before blastulation, indicating the possibility of achromosomal cleavage because of the absence of DNA synthesis. Based on the above, it is concluded that these analogues can be used as other inhibitors of eukaryotic DNA synthesis and DNA polymerase alpha.

Animals↗
PubMed1983

Inhibition of DNA polymerases of sea urchin by palmitoyl coenzyme A.

Palmitoyl CoA noncompetitively inhibited the activities of DNA polymerase alpha and gamma, prepared from sea urchin germ cells, with Ki values of 28 microM and 116 microM, respectively. Myristoyl CoA also inhibited DNA polymerase alpha and gamma, while coenzyme A, short chain fatty acyl CoA's, Na-myristate and Na-palmitate failed to inhibit the enzymes. It was concluded that both the long hydrocarbon chain and CoA moiety of long chain fatty acyl CoA's are necessary for inhibition of DNA polymerase activity. DNA polymerase beta was not inhibited by long chain fatty acyl CoA's.

Acyl Coenzyme A↗