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M Nakatsuka

Publications and source records attributed to M Nakatsuka.

At least 37 records · Page 2Linked to original sources
PubMed1999

Isolation and characterization of the distal promoter region of mouse Cbfa1.

Cbfa1 is an essential transcription factor for bone formation, and as such little is known about the region responsible for the transcriptional regulation of this gene. Here we report the determination of the transcription start sites, isolation and partial characterization of distal promoter region of this gene. Three transcription start sites were identified by the 5'-Cap site method, recently invented for rapid examination of the 5'-end of genes of interest. A reporter construct containing 1.8 kb of 5' of transcription start sites had approximately 25-fold more luciferase activity than the promoter-less vector in osteoblastic cell lines. Deletion analysis of the reporter construct demonstrated that the minimal region to express promoter activity lies between bp -168 and -99, taking the most downstream transcription start site as +1. By Northern blot analysis, mRNA expression from the distal promoter was detected in the differentiated osteoblastic cell lines, UMR-106, ROS17/2.8 and MC3T3-E1, but not in cell lines of immature phenotype or originated from other organs. Luciferase activity was strongest in UMR-106 and ROS17/2.8, and weakest in COS-1 and HepG2, which are cell lines originating from other organs, corresponding to the level of mRNA expression. These results demonstrated that the distal promoter region examined here is important for tissue- and cell-type-specific gene expression of Cbfa1.

Animals↗
PubMed1999

Cbfa1 isoforms exert functional differences in osteoblast differentiation.

Cbfa1 is an essential transcription factor for osteoblast differentiation and bone formation. We investigated functional differences among three isoforms of Cbfa1: Type I (originally reported as Pebp2alphaA by Ogawa et al. (Ogawa, E., Maruyama, M., Kagoshima, H., Inuzuka, M., Lu, J., Satake, M., Shigesada, K., and Ito, Y. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 6859-6863), Type II (originally reported as til-1 by Stewart et al. (Stewart, M., Terry, A., Hu, M., O'Hara, M., Blyth, K., Baxter, E., Cameron, E., Onions, D. E., and Neil, J. C. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 8646-8651), and Type III (originally reported as Osf2/Cbfa1 by Ducy et al. (Ducy, P., Zhang, R., Geoffroy, V., Ridall, A. L., and Karsenty, G. (1997) Cell 89, 747-754). A reverse transcriptase-polymerase chain reaction analysis demonstrated that these isoforms were expressed in adult mouse bones. The transient transfection of Type I or Type II Cbfa1 in a mouse fibroblastic cell line, C3H10T1/2, induced the expression of alkaline phosphatase (ALP) activity. This induction was synergistically enhanced by the co-introduction of Xenopus BMP-4 cDNA. In contrast, the transient transfection of Type III cDNA induced no ALP activity. In C3H10T1/2 cells stably transfected with each isoform of Cbfa1, the gene expression of ALP was also strongly induced in cells transfected with Type I and Type II Cbfa1 but not in cells with Type III Cbfa1. Osteocalcin, osteopontin,and type I collagen gene expressions were induced or up-regulated in all of the cells stably transfected with each isoform of Cbfa1, and Type II transfected cells exhibited the highest expression level of osteocalcin gene. A luciferase reporter gene assay using a 6XOSE2-SV40 promoter (6 tandem binding elements for Cbfa1 ligated in front of the SV40 promoter sequence), a mouse osteocalcin promoter, and a mouse osteopontin promoter revealed the differences in the transcriptional induction of target genes by each Cbfa1 isoform with or without its beta-subunit. These results suggest that all three of the Cbfa1 isoforms used in the present study are involved in the stimulatory action of osteoblast differentiation, but they exert different functions in the process of osteoblast differentiation.

Animals↗
PubMed1999

Generation of peroxynitrite and apoptosis in placenta of patients with chorioamnionitis: possible implications in placental abruption.

The reaction of nitric oxide (NO) and superoxide results in the formation of peroxynitrite, a potent and relatively long-lived oxidant. In infectious diseases, these molecules are not only bactericidal but also toxic to host cells. Chorioamnionitis is often complicated by premature rupture of membranes and can be associated with placental abruption. These diseases are significant causes of premature low-birth-weight deliveries and consequently the morbidity and mortality of neonates. Lipopolysaccharide, bacterial endotoxin, is known to be elevated in the amniotic fluid of patients with chorioamnionitis. Lipopolysaccharide is known to induce the formation of NO and superoxide. We report here that nitrite/nitrate, stable metabolites of NO, were increased in serum from patients with chorioamnionitis. Immunohistochemical studies demonstrated enhanced expression of inducible NO synthase and formation of nitrotyrosine, a footprint of peroxynitrite, in the placentae from patients with chorioamnionitis and also in patients with placental abruption. Furthermore, apoptotic cell death was also increased in the placentae from patients with both diseases. These results suggest that chorioamnionitis and a portion of placental abruption may share a common cascade of placental injury. Nitric oxide and its metabolities may play an important role in this cascade.

Adult↗
PubMed1999

Clinical experience of long-term transdermal treatment with nitric oxide donor for women with preeclampsia.

Isosorbide dinitrate (ISDN), a nitric oxide donor, was applied transdermally for 4-16 days to 4 preeclamptic women with oligohydramnios, intrauterine fetal growth retardation (IUGR), and elevated resistance of blood flow in the uterine arteries. Pulsed Doppler ultrasonography revealed immediate and drastic improvement of pulsatility index (PI) of uterine arteries following treatment with ISDN. The average PI in uterine arteries of the 4 patients was reduced to approximately 67% of that of the untreated state. In 2 patients the amniotic fluid gradually increased over a few days which suggested improvement of fetoplacental circulation during administration of ISDN. This study suggests that long-term transdermal ISDN is an effective therapy, at least in a portion of preeclamptic women, to avoid maternal hypertension, fetal distress, oligohydramnios, and IUGR, and consequentially to prolong the gestational period.

Administration, Cutaneous↗
PubMed1998

Cloning of rabbit TR4 and its bone cell-specific activity to suppress estrogen receptor-mediated transactivation.

To clone a new nuclear receptor, we screened a rabbit heart complementary DNA (cDNA) library with degenerate oligonucleotide probes corresponding to the DNA-binding domain of nuclear receptors, which is highly conserved among receptors. One of the cDNA clones, clone 23, encodes a novel protein of 596 amino acids, and predicted molecular mass is 66 kDa. Homology search analysis identified this protein as rabbit TR4 (TR4-0). We also cloned the cDNA encoding a rabbit TR4 isoform (TR4-1), which lacks the putative C-terminal ligand-binding domain (350 amino acids) caused by a 23-bp exon deletion, which probably occurred during messenger RNA (mRNA) splicing. Northern blot analysis showed that TR4s are expressed with two kinds of mRNAs (9.0 kb and 2.8 kb), both of which are relatively abundant in brain, testis, and bone. RT-PCR analysis, using pairs of primers specific for each TR4, showed that both types of receptor express in various tissues. Furthermore, both are present in primary osteoblasts and bone marrow cells, though the mRNA levels of TR4-0 were much higher than those of TR4-1. A functional study, using a transient transfection assay, showed that both receptors suppressed retinoid X receptor (RXR)-retinoid acid receptor, RXR-TR, and RXR-VDR-mediated transactivation significantly in COS-1 and osteosarcoma cells (UMR-106, ROS17/2.8) and that TR4-0 was much more effective than TR4-1. Unexpectedly, we found that the TR4s effectively suppressed estrogen receptor-mediated transactivation in bone cells, but neither in kidney (COS-1) nor breast cancer cells (MCF-7, one of the major target cells of the estrogen action). Thus, the present study shows a novel property of the TR4 orphan receptor, acting as a bone cell-specific repressor in the estrogen receptor-mediated signaling pathway.

Amino Acid Sequence↗
PubMed1998

Metabolism-based inactivation of penile nitric oxide synthase activity by guanabenz.

Guanabenz (Wytensin) was shown to inactivate nitric oxide synthase (NOS) activity in vitro and in vivo. In in vitro studies with the use of a cytosolic fraction from penile tissue, the inactivation was found to depend on NADPH, time, and the concentration of guanabenz. The L-, but not the D-, isomer of arginine could protect from the inactivation, suggesting an active site-directed event. The kinetics of inactivation could be described by an apparent dissociation constant for the initial reversible complex (Ki) and a pseudo first-order inactivation constant (kinact) of 38.5 microM and 0.179 min-1, respectively. In in vivo studies, guanabenz was shown to inhibit penile cytosolic NOS activity in a dose- and time-dependent manner. Treatment of rats with guanabenz (5 mg/kg/day) for 4 days caused a decrease of approximately one-half in the NOS activity of the penile cytosolic fraction with a concomitant loss in the amount of immunodetectable NOS protein. Treatment for 4 days at a dose of 0. 5 mg/kg/day showed a similar decrease in activity, whereas a dose of 0.05 mg/kg/day showed no effects. Due to the multitude of processes that are regulated by NO, the inactivation of NOS is a potential mechanism to be considered in a variety of biological effects associated with drugs.

Animals↗
PubMed1997

Cloning of mouse FGF10 and up-regulation of its gene expression during wound healing.

We cloned the mouse homolog of FGF10, which was recently reported as a new member of the FGF family. The predicted molecular mass of this molecule is 23.6 kDa, and both nucleotide and amino acid sequences show high degrees of similarity with those of the rat. Examination of mouse FGF10 mRNA expression in various tissues and developmental stages by Northern hybridization revealed tissue- and developmental stage-specific expression of the gene. Similarly to the rat counterpart, mouse FGF10 mRNA (4.5 kb) was expressed relatively abundantly in embryos and the lung, and at much lower levels in brain and heart. In addition, a shorter transcript (1.3 kb) is expressed only in testis. Considering the high similarity in primary structure between FGF10 and FGF7 (known as keratinocyte growth factor; KGF), we also examined the gene expression of FGF10 during wound healing using a mouse model. FGF10 mRNA was highly induced 1 day after injury and decreased rapidly by 3 days. This suggests that FGF10 is a primary factor in the process of wound healing similarly to other growth factors such as TGF alpha and FGF7.

Amino Acid Sequence↗
PubMed1997

Structure and expression of human fibroblast growth factor-10.

We isolated the cDNA encoding a novel member of the human fibroblast growth factor (FGF) family from the lung. The cDNA encodes a protein of 208 amino acids with high sequence homology (95.6%) to rat FGF-10, indicating that the protein is human FGF-10. Human FGF-10 as well as rat FGF-10 has a hydrophobic amino terminus ( approximately 40 amino acids), which may serve as a signal sequence. The apparent evolutionary relationships of human FGFs indicate that FGF-10 is closest to FGF-7. Chromosomal localization of the human FGF-10 gene was examined by in situ hybridization. The gene was found to map to the 5p12-p13 region. Human FGF-10 (amino acids 40 to 208 with a methionine residue at the amino terminus) was produced in Escherichia coli and purified from the cell lysate. Recombinant human FGF-10 (approximately 19 kDa) showed mitogenic activity for fetal rat keratinizing epidermal cells, but essentially no activity for NIH/3T3 cells, fibroblasts. The specificity of mitogenic activity of FGF-10 is similar to that of FGF-7 but distinct from that of bFGF. In structure and biological activity, FGF-10 is similar to FGF-7.

Amino Acid Sequence↗
PubMed1997

Increased intra-abdominal pressure and cardiac filling pressures in obesity-associated pseudotumor cerebri.

OBJECTIVES: To determine whether intra-abdominal pressure (as estimated from urinary bladder pressure) is elevated in patients with central obesity (as measured by sagittal abdominal diameter) and pseudotumor cerebri and whether this increased intra-abdominal pressure is associated with increased pleural pressure and cardiac filling pressure, implying a resistance to venous return from the brain. DESIGN: Nonrandomized, prospective. SETTING: University hospital, operating room. MAIN OUTCOME MEASUREMENTS: Intracranial pressure, urinary bladder pressure, sagittal abdominal diameter, transesophageal pleural pressure, central venous pressure, pulmonary artery pressure, and pulmonary artery occlusion pressure. SUBJECTS: Six women with pseudotumor cerebri (one with CSF leak, one with lumboperitoneal shunt). RESULTS: Urinary bladder pressure (22 +/- 3 cm H2O) and sagittal abdominal diameter (29 +/- 3 cm) were significantly elevated in these patients with elevated intracranial pressure (293 +/- 80 mm H2O) compared with a previously reported group of nonobese control patients. The transesophageal pleural pressure (15 +/- 10 mm Hg), central venous pressure (20 +/- 6 mm Hg), mean pulmonary artery pressure (31 +/- 6 mm Hg), and pulmonary artery occlusion pressure (21 +/- 7 mm Hg) were all markedly elevated compared with published normal values and with previous data from obese patients without pseudotumor cerebri. CONCLUSIONS: These data support the hypothesis that central obesity raises intra-abdominal pressure, which increases pleural pressure and cardiac filling pressure, which impede venous return from the brain, leading to increased intracranial venous pressure and increased intracranial pressure associated with pseudotumor cerebri.

Abdomen↗
PubMed1997

Quantification of area at risk in acute myocardial infarction by tomographic imaging.

UNLABELLED: The purpose of this study was twofold: to validate, in a phantom heart model, a simple threshold technique for the quantification of defect size using 123I-15-(p-iodophenyl)-3-(R,S)-methyl pentadecanoic acid (BMIPP) imaging and to compare, in patients with acute myocardial infarction, defect size as shown by BMIPP imaging, with the extent of severe hypokinesis shown by left ventriculography. METHODS: In a phantom study, defect size was calculated using a standard geometric formula. In a clinical study, BMIPP imaging was performed in 20 patients 10 +/- 5 days after the onset of their infarction. Using the centerline method, the area at risk was defined by contrast ventriculography as the percentage of chords with wall motion >2 s.d. below normal. RESULTS: In the phantom study, a threshold value of 60% yielded the best agreement between true and measured defect size. In the clinical study, the defect size shown by BMIPP imaging was greater in anterior than in inferior infarcts (p < 0.001) and correlated well with the risk area revealed by contrast ventriculography (r = 0.80, p < 0.0001) CONCLUSION: The above preliminary data, admittedly from a small group of patients, suggest that tomographic BMIPP imaging provides an accurate quantification of defect size by means of a simple threshold technique and, in the subacute phase, permits determination of the amount of myocardium at risk after acute myocardial infarction.

Coronary Angiography↗
PubMed1996

Pulmonary vascular resistance and right ventricular function in morbid obesity in relation to gastric bypass surgery.

STUDY OBJECTIVE: To investigate right ventricular function (RVF) during gastric bypass surgery in morbidly obese patients. To study the influence of obstructive sleep apnea syndrome on hemodynamics and RVF as a preoperative evaluation in morbidly obese patients. DESIGN: Prospective study. SETTING: Surgical patients at a university hospital. PATIENTS: 14 morbidly obese patients undergoing gastric bypass surgery. INTERVENTIONS: Hemodynamic changes and RVF were investigated using a thermodilution ejection fraction volumetric catheter and the REF-1 computer. MEASUREMENTS AND MAIN RESULTS: Hemodynamic parameters and RVF were measured: (1) before induction of anesthesia, awake, (2) after induction of anesthesia, (3) after opening the abdomen, and (4) after closing the abdomen. Esophageal pressure was measured in the midesophagus after induction of anesthesia with a camino catheter and its device. Morbidly obese patients with obstructive sleep apnea syndrome had significantly lower PaO2, higher PaCO2, and higher pulmonary artery (PA) pressure and pulmonary vascular resistance (PVR) compared with patients without sleep apnea (p < 0.05). However, there were no significant differences in heart rate, mean blood pressure, pulmonary capillary wedge pressure (PCWP), right ventricular ejection fraction (RVEF), and right ventricular end-diastolic volume. During gastric bypass procedure, PA pressure, PCWP, and PVR decreased significantly after opening the abdomen (p < 0.05). There were no significant changes in RVEF and cardiac index during gastric bypass procedure. CONCLUSION: The presence of chronic hypoxemia and hypercarbia in our morbidly obese patients with obstructive sleep apnea syndrome while awake, causes significant increases in PA pressure and PVR. We also demonstrated that RVEF did not change significantly during gastric bypass procedure despite significant decreases in PA pressure, PCWP, and PVR after opening the abdomen. This decrease in PA pressure and PVR may be caused by decreases in pleural pressure reflected by a concomitant decrease in esophageal pressure.

Adult↗
PubMed1995

1988: unilateral high-frequency jet ventilation during one-lung ventilation for thoracotomy. Updated in 1995.

One-lung ventilation is indicated during thoracic operations for bronchopleural fistula, pulmonary abscess, and pulmonary hemorrhage in spite of the possibility of the development of severe hypoxemia. To evaluate methods for improving oxygen transport during one-lung ventilation, we applied high-frequency jet ventilation (HFJV) and continuous positive airway pressure (CPAP) to the nondependent lung following deflation to atmospheric pressure in each procedure, and measured the effects on cardiac output and arterial oxygenation. In each case, the dependent lung was ventilated with conventional intermittent positive pressure ventilation (IPPV). Eight patients were studied during posterolateral thoracotomy using double-lumen endobronchial tubes. HFJV or CPAP to the nondependent lung improved arterial oxygenation significantly during both closed and open stages of the surgical procedures (p < 0.008). When the chest was open, HFJV maintained satisfactory cardiac output, whereas CPAP usually decreased cardiac output (p < 0.008). There were no significant differences in mean partial pressure of arterial carbon dioxide between HFJV, CPAP, and deflation to atmospheric pressure. In conclusion, HFJV to the nondependent lung provides not only satisfactory oxygenation but also good cardiac output, thereby maintaining better oxygen transport than CPAP or deflation to atmospheric pressure, while the dependent lung is ventilated with IPPV during one-lung ventilation for thoracotomy.

High-Frequency Jet Ventilation↗
PubMed1994

Selective inhibition of the 12-lipoxygenase pathway of arachidonic acid metabolism by L-arginine or sodium nitroprusside in intact human platelets.

L-arginine (1-100 microM) or sodium nitroprusside (1-100 microM) caused a concentration dependent decrease in the metabolism of exogenously added arachidonic acid via the 12-lipoxygenase pathway in intact human platelets, as determined by the use of an HPLC assay. NG-monomethyl-L-arginine, but not the D-isomer of this compound, diminished the inhibitory effect of L-arginine. The D isomer of arginine had no effect. The cyclooxygenase pathway was much less susceptible to these effects. This study indicated that nitric oxide formed in intact human platelets selectively inhibited 12-lipoxygenase over that of cyclooxygenase and suggests that such inhibition may be an important regulatory mechanism.

Arachidonate 12-Lipoxygenase↗