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Biomedical subjects

Y Nishimoto

Publications and source records attributed to Y Nishimoto.

At least 55 records · Page 3Linked to original sources
PubMed1997

Identification of CFDD (common regulatory factor for DNA replication and DREF genes) and role of its binding site in regulation of the proliferating cell nuclear antigen gene promoter.

The Drosophila proliferating cell nuclear antigen (PCNA) gene promoter contains at least three transcriptional regulatory elements, the URE (upstream regulatory element), DRE (DNA replication-related element), and E2F recognition sites. In nuclear extracts of Drosophila Kc cells, we detected a novel protein factor(s), CFDD (common regulatory factor for DNA replication and DREF genes) that appeared to recognize two unique nucleotide sequences (5'-CGATA and 5'-CAATCA) and bind to three sites in the PCNA gene promoter. These sites were located at positions -84 to -77 (site 1), -100 to -93 (site 2) and -134 to -127 (site 3) with respect to the transcription initiation sites. Sites 2 and 3 overlapped with DRE and URE, respectively, and the 5'-CGATA matched with the reported recognition sequence of BEAF-32 (boundary element-associated factor of 32 kDa). Detailed analyses of CFDD recognition sequences and experiments with specific antibodies to DREF (DRE-binding factor) and BEAF-32 suggest that CFDD is different from DREF, UREF (URE-binding factor) and BEAF-32. A UV cross-linking experiment revealed that polypeptides of approximately 76 kDa in the nuclear extract interact directly with the CFDD site 1 sequence. Transient expression assays of chloramphenicol acetyltransferase (CAT) in Kc cells transfected with PCNA promoter-CAT fusion genes carrying mutations in CFDD site 1 and examination of lacZ expression from PCNA promoter-lacZ fusion genes carrying mutations in site 1, introduced into flies by germ line transformation, revealed that CFDD site 1 plays an important role for the promoter activity both in cultured cells and in living flies. In addition to the PCNA gene, multiple CFDD sites were found in promoters of the DNA polymerase alpha and DREF genes, and CFDD binding to the DREF promoter was confirmed. Therefore, CFDD may play important roles in regulation of Drosophila DNA replication-related genes.

Amino Acid Sequence↗
PubMed1997

Right ventricular myxoma: report of a case.

We herein report a 36-year-old man who underwent surgical resection for myxoma. Preoperative two-dimensional echocardiography demonstrated a mass in the right ventricle. Intraoperatively, the tumor was found to derive from an anterior papillary muscle of the tricuspid valve. The tumor was successfully excised and the tricuspid valve was repaired with chordoplasty and annuloplasty. A histopathological examination revealed myxoma and a 2-year follow-up has shown no evidence of recurrence or tricuspid valve regurgitation.

Adult↗
PubMed1997

Extrahepatic portosystemic venous shunt without portal hypertension.

A 72-year-old woman was admitted for recurrent episodes of encephalopathy. Laboratory data showed mild liver dysfunction and hyperammonemia, while she had neither anemia nor splenomegaly. The dilated inferior mesenteric vein (IMV) was opacified retrogradely from the superior mesenteric vein by superior mesenteric arteriography, and IMV was found to connect with the inferior vena cava (IVC) through a torturous shunt. No obstruction of the extrahepatic portal vein or hepatic vein was observed by arteriography. Histological evaluation of the liver biopsy indicated remarkable fatty change without cirrhosis. Finally, we diagnosed this case as extrahepatic portosystemic venous shunt without portal hypertension.

Aged↗
PubMed1997

Protein synthesis dependent and independent apoptosis of murine splenic T cells.

To elucidate the mechanism of apoptosis in matured T cells, we examined the effect of phorbol esters capable of activating protein kinase C, cAMP agonists, calcium ionophores, and dexamethasone on apoptosis in mouse splenic T cells. All of these agents induced apoptosis in thymocytes. Neither phorbol esters nor calcium ionophores induced apoptosis in the splenic T cells. Dibutyrylcyclic AMP induced apoptosis in splenic T cells but to a lesser extent than that in thymocytes. The results suggest that matured T cells are more resistant to various apoptogenic stimuli than thymocytes. Dexamethasone induced apoptotic cell death in splenic T cells as well as in thymocytes. Cycloheximide, an inhibitor of protein synthesis which completely inhibited thymocyte apoptosis, induced slight apoptosis in splenic T cells, but it inhibited the dexamethasone-induced apoptosis to a level at which cycloheximide induced apoptosis, indicating that at least two pathways of apoptosis exist in peripheral T cells; one protein synthesis dependent and the other independent. Taken together, the results suggest that the apoptosis of peripheral T cells is regulated by a mechanism different from that of thymocytes.

Animals↗
PubMed1996

Isolation and characterization of cDNA for DREF, a promoter-activating factor for Drosophila DNA replication-related genes.

DREF, a transcription regulatory factor which specifically binds to the promoter-activating element DRE (DNA replication-related element) of DNA replication-related genes, was purified to homogeneity from nuclear extracts of Drosophila Kc cells. cDNA for DREF was isolated with the reverse-transcriptase polymerase chain reaction method using primers synthesized on the basis of partial amino acid sequences and following screening of cDNA libraries. Deduced from the nucleotide sequences of cDNA, DREF is a polypeptide of 701 amino acid residues with a molecular weight of 80,096, which contains three characteristic regions, rich in basic amino acids, proline, and acidic amino acids, respectively. Deletion analysis of bacterially expressed DREF fused with glutathione S-transferase (GST-DREF) indicated that a part of the N-terminal basic amino acid region (16-115 amino acids) is responsible for the specific binding to DRE. A polyclonal and four monoclonal antibodies were raised against the GST-DREF fusion protein. The antibodies inhibited specifically the transcription of DNA polymerase alpha promoter in vitro. Cotransfection experiments using Kc cells demonstrated that overproduction of DREF protein overcomes the repression of the proliferating cell nuclear antigen gene promoter by the zerknüllt gene product. These results confirmed that DREF is a trans-activating factor for DNA replication-related genes. Immunocytochemical analysis demonstrated the presence of DREF polypeptide in nuclei after the eighth nuclear division cycle, suggesting that nuclear accumulation of DREF is important for the coordinate zygotic expression of DNA replication-related genes carrying DRE sequences.

Amino Acid Sequence↗
PubMed1996

Depth dependence of the perturbing effect of placing a bulky group (oxazolidine ring spin labels) in the membrane on the membrane phase transition.

Electron paramagnetic resonance (EPR) and differential scanning calorimetry (DSC) have been used to study the effect on the phase transition of dimyristoylphosphatidylcholine membranes of incorporating various stearic acid spin labels (SASL's) that contain the bulky oxazolidine ring at various positions along the stearyl chain. SASL's lowered the phase transition temperature and decreased the size of the cooperative unit, with the effects stronger in the order of 9- > 12- > 5- > 16-SASL > stearic acid (no label). Incorporation of stearic acid without the spin label slightly increases the phase transition temperature. Incorporation of 9-SASL (3 mol% of lipid) decreased the transition temperature by 1.8 degrees C and the cooperative unit to 1/5 of that without the spin label, while the effect of 16-SASL was slight. The effect on transition enthalpy was small. It is concluded that the perturbing effect of placing a bulky group on the alkyl chain on phase transition is through inducing packing defects in the gel-phase.

Calorimetry, Differential Scanning↗
PubMed1996

[Rupture of thoracic aorta due to blunt trauma: a report of three cases].

Three cases of the thoracic aortic rupture due to blunt trauma were successfully treated. All of the cases were young male motorcyclists. The chest X-ray on admission showed either a widened mediastinum or an apical extrapleural cap sign. Ruptures were confirmed by an enhanced CT clearly showing the presence of a psuedoaneurysm and a hematoma around the isthmus of the aorta. One of them was operated upon urgently and others in whom heparin could not be used because of associated injuries were operated upon electively two and three months after the traffic accident, respectively. In two of them, aneurysms were removed and replaced with dacron grafts and in the remaining one, the aneurysm was resected and repaired directly under the femoro-femoral bypass. Post-operative courses were uneventful. They discharged and are doing well after the treatment for associated injuries. In view of the high early mortality of aortic rupture, an early diagnosis and treatment is important. But in the case who has stable hemodynamics and contraindication for heparinization, a delayed operation may be recommended.

Accidents, Traffic↗
PubMed1995

Dynamic interactions of rabbit liver cytochromes P450IA2 and P450IIB4 with cytochrome b5 and NADPH-cytochrome P450 reductase in proteoliposomes.

Purified liver microsomal cytochrome P450IA2 or P450IIB4 was co-reconstituted with cytochrome b5 or NADPH-cytochrome P450 reductase in phosphatidylcholine-phosphatidylethanolamine-phosphatidylserine vesicles at a lipid to P450 weight ratio of 2 by cholate dialysis procedures. The proteoliposomes catalyzed drug oxidation. Rotational diffusion of cytochrome P450 was measured by observing the decay of absorption anisotropy, r(t), after photolysis of the heme.CO complex. Analysis of r(t) was based on a "rotation-about-membrane normal" model. The absorption anisotropy decayed within 1 ms to a time-independent value, r3. Different rotational mobility for the two cytochrome P450s was observed. Though 20% of cytochrome P450IA2 was immobile, all cytochrome P450IIB4 molecules were rotating. The rotational relaxation time, phi, of the mobile population was 237 microseconds for cytochrome P450IA2 and 160 microseconds for cytochrome P450IIB4. The two cytochrome P450s have shown very different interactions with cytochrome b5 and NADPH-cytochrome P450 reductase. By the presence of the redox partner, the mobile population of cytochrome P450IA2 was increased significantly from 80% to 96% (plus cytochrome b5) and to 89% (plus NADPH-cytochrome P450 reductase) due to dissociation of P450 oligomers. On the other hand, the mobility of cytochrome P450IIB4 was not considerably affected by the presence of cytochrome b5 or NADPH-cytochrome P450 reductase as judged by little difference in phi and r3, keeping the mobile population of 100%. These results imply that cytochrome P450IA2 forms a transient association with cytochrome b5 and NADPH-cytochrome P450 reductase.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

A nucleotide sequence essential for the function of DRE, a common promoter element for Drosophila DNa replication-related genes.

Promoter regions of the Drosophila proliferating cell nuclear antigen (PCNA) gene and the DNA polymerase alpha 180-kDa catalytic subunit gene contain a common 8 base pair (bp) promoter element, 5'-TATCGATA (DRE, Drosophila DNA replication-related element). We have generated various base substitutions and internal deletions in and around DRE (nucleotide positions -93 to -100 with respect to the transcription initiation site) of the PCNA gene in vitro and subsequently examined their effects on the binding to DREF (DRE-binding factor) and PCNA gene promote activity in cultured Drosophila Kc cells as well as in living flies. Gel mobility shift assays using nuclear extracts of Kc cells with and without competitor DNA fragments carrying the mutations indicated that the 10-bp sequence from positions -91 to -100 is essential for complex formation with DREF. Transient expression assays of chloramphenicol acetyl-transferase (CAT) in Kc cells transfected with PCNA promoter-CAT fusion genes carrying the mutations revealed that the 8-bp sequence from -93 to -100 is essential for activation of the promoter in Kc cells. Examination of lacZ expression from PCNA promoter-lacZ fusion genes carrying the mutations, introduced into flies by germ-line transformation, revealed that the 8-bp sequence is also important for DRE function during development. However, we obtained two exceptional mutations in the 8-bp sequence that did not or only marginally affected the PCNA gene promoter activity in transgenic flies. Both of these mutations effectively reduced the promoter activity in CAT transient expression assay in Kc cells and the binding to DREF in vitro. Therefore, the 8-bp sequence requirement for DRE function appears to be less stringent in living flies than in the cultured cell or in vitro cases.

Animals↗
PubMed1995

Expression patterns of DNA replication enzymes and the regulatory factor DREF during Drosophila development analyzed with specific antibodies.

Specific antibodies were prepared against Drosophila DNA polymerase epsilon and DREF, a regulatory factor for DNA replication-related genes. Using these antibodies together with those for DNA polymerase alpha and proliferating cell nuclear antigen (PCNA), we examined expression patterns and sub-cellular distributions of these proteins during Drosophila development. DNA polymerase alpha, epsilon and PCNA proteins were maternally stored in unfertilized eggs and maintained at high levels during embryogenesis. With distinct nuclear localization, proteins were observed in embryos at interphase stages throughout the 13 nuclear division cycles, suggesting that they all participate in rapid nuclear DNA replication during these cycles. In contrast, maternal storage of a DREF protein was relatively low and its level increased throughout embryogenesis. Strong nuclear staining with the anti-DREF antibody was not observed until the nuclear division cycle 8. Immunostaining of various larval tissues from transgenic flies carrying the PCNA gene promoter-lacZ fusion gene revealed co-expression of DREF, PCNA and lacZ, suggesting that DREF regulates the expression of PCNA gene in these tissues. In addition, we detected a relatively high level of DREF in adult males as well as females. Since DNA polymerase alpha, epsilon and PCNA are hardly detectable in adult males, DREF very likely regulates genes other than those closely linked to DNA replication in adult males.

Animals↗
PubMed1995

Distribution of PCNA during postblastoderm cell division cycles in the Drosophila melanogaster embryo: effect of a string- mutation.

We used immunocytochemical methods and a specific antibody to identify proliferating cell nuclear antigen (PCNA) in Drosophila embryos during the postblastoderm cell division cycles. The strong nuclear staining observed during interphase disappeared at prophase, staining remained nil throughout the remaining mitotic (M) phases and reappeared in nuclei at the next interphase. As the embryos with the homozygous string- mutation sustained the strong staining signal in nuclei throughout embryonic development, disappearance of the staining signal probably depends on string function and is coupled with the onset of mitosis. When cells in embryos were arrested at M phase following treatment with TN-16 or Vinblastine, the staining signal with the anti-PCNA antibody was lost. However, Western blots showed that the level of PCNA protein in M phase-arrested embryos did not decrease. Therefore, the disappearance of staining signals is apparently due to reorganization of PCNA protein in the multiprotein complex in nuclei, rendering it inaccessible to the antibody, rather than to the degradation of PCNA protein. In contrast to findings in developing embryos, cultured Drosophila Kc cells stained strongly with the anti-PCNA antibody, during both interphase and the M phase. In genetic crossing experiments of transgenic flies carrying the lacZ gene under the control of the PCNA gene regulatory region (-607 to +137 with respect to the transcription initiation site) with string- mutant files, the PCNA gene promoter seems to function in a manner independent of cell cycle progression or of functions of the string gene.

Animals↗
PubMed1994

Crystallization of 31 kDa C-terminal fragment of rat DNA polymerase beta.

A 248 residue C-terminal fragment of rat DNA polymerase beta (335 amino acid residues), a eukaryotic DNA repair enzyme, has been crystallized from polyethylene glycol 6000 solution. The crystals are orthorhombic, space group P2(1)2(1)2 with cell dimensions a = 120.3 A, b = 64.2 A, c = 39.4 A, and contain a single 31 kDa fragment in an asymmetric unit. The crystals diffract to 2.8 A resolution with laboratory X-ray source, and to 2.3 A resolution with synchrotron X-ray source, and are suitable for detailed structural analysis.

Animals↗
PubMed1994

Neuroendocrine studies in dementia patients: responses of plasma GH and PRL following bromocriptine administration.

Bromocriptine stimulates growth hormone (GH) secretion at the hypothalamus and suppresses prolactin (PRL) secretion at the pituitary level. We administered bromocriptine to 30 patients with dementia of the Alzheimer type (DAT), 30 patients with multi-infarct dementia (MID) and 22 age matched healthy controls, and compared response patterns of GH and PRL. Incomplete PRL suppressive responses (suppression rate < 50%) were seen in 36.7% of DAT patients and in 30.0% of MID patients, indicating that both groups had the same degree of pituitary dysfunctions. Blunted GH responses (< 5 ng/ml) were seen in 93.3% of DAT patients, in 63.3% of MID patients and in 31.8% of the controls. The results indicate that neuroendocrine regulation of GH is more selectively and severely damaged in DAT patients than in MID patients at the hypothalamus.

Aged↗
PubMed1994

[Trial of self-management of outpatients with implanted reservoir for arterial infusion of anticancer drugs].

We attempted to instruct the outpatients with malignant liver tumor (metastatic liver cancer and advanced hepatocellular carcinoma) who received intermittent arterial infusion chemotherapy using implanted reservoir to detach the devices for arterial infusion by themselves at home. All patients who received our instruction could master self-detachment at home, which shortened the hospitalized day. And patients' desire for this treatment promoted with improvement of the recognition of their families to participate in treatment together. We experienced no trouble which disturb the treatment. This methods was considered to promote the improvement of QOL of these patients.

Ambulatory Care↗
PubMed1994

Relationship between sigma-like site and progesterone-binding site of adult male rat liver microsomes.

An increasing amount of evidence suggests that the sigma (sigma) sites, putative targets for a variety of psychotomimetic and antipsychotic drugs, exist not only in the brain but also in various peripheral organs. However, there are many ambiguities as to their biological roles, subcellular distributions, endogenous ligands and so on. We therefore performed our study for clarification of some of these ambiguities. As a result, we demonstrated that adult male rat liver microsomes, especially smooth endoplasmic reticulum, possessed a saturable haloperidol-binding site closely resembling the sigma site, with a high affinity (Kd 1.0 +/- 0.3 nmol/L) and high capacity (Bmax 9.3 +/- 1.5 pmol/mg protein) and with the rank order of affinity of the ligands: haloperidol > reduced haloperidol > clorgyline > ifenprodil > 1,3-di(2-tolyl)guanidine, (-)-butaclamol > GBR-12909 > SKF-525A > progesterone > 5 alpha-dihydrotestosterone > R(+)-3- (hydroxyphenyl)-N-propylpiperidine > testosterone >> corticosteroids, estradiol-17 beta, cholesterol and neuroactive compounds displaying high affinities for other neurotransmitter receptors such as dopamine D2, serotonin (5-HT1A and 5-HT2) and alpha 1-adrenergic and GABAA receptors. This rank order showed a high correlation (r = 0.908) with that of a large portion (approximately 85%) of specific progesterone-binding site (Kd 31.0 +/- 3.5 nmol/L, Bmax 5.7 +/- 0.2 pmol/mg protein) of the same source. Therefore, these two sites were suggested to be the same or closely related. Furthermore, we provide a strong suggestion that these sites neither are identical with some subforms of the microsomal cytochromes P-450 or other steroid/drug-metabolizing enzymes nor participate universally and directly in the progesterone-metabolizing processes.

Animals↗
PubMed1994

[The causes and timing of reoperation for coronary artery disease].

In eight hundred and sixteen consecutive patients undergoing coronary artery bypass grafting (CABG) within the past 19 years (1974 to 1993), repeated CABG were performed on 12 patients (1.47%). The main reasons for repeated CABG were graft failure (GF), progression of native disease (NP) and incomplete revascularization (IR). The incidence of GF only and GF combined NP were high either within a half year or around 5 years after CABG. When the recurrence of angina is noted after CABG, the catheter intervention should be chosen at first but the reoperation should be done not so late. Although all patients survived from reoperation, three patients continued to have mild angina pectories. If a reoperation is inevitable, maximum utilization of arterial graft and accomplishment of complete revascularization are emphasized.

Aged↗
PubMed1994

[Reoperation for mitral valve disease].

From 1978 to March 1994, 95 patients underwent repeated open heart surgery for mitral valve diseases. There were 48 men and 47 women. The mean age was 50 and the mean interval between initial and second operation was 8 years. The initial operations were OMC in 39, MAP in 13, OMC+MAP in 11, MVR in 30 and AVR in 2 patients. The hospital death occurred in 31 cases (32.6%). In recent cases, however the hospital death were reduced to 4 (14.8%). This was attributed to the change in myocardial protection from previous intermittent antegrade crystalloid to recent continuous combined antegrade and retrograde cardioplegia.

Adolescent↗