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K Nose

Publications and source records attributed to K Nose.

At least 73 records · Page 4Linked to original sources
PubMed1995

Isolation of a novel ras-recision gene that is induced by hydrogen peroxide from a mouse osteoblastic cell line, MC3T3-E1.

Hydrogen peroxide appears to mediate growth factor actions, and it inhibits DNA synthesis in normal mouse osteoblastic cells (MC3T3-E1) at non-toxic doses. However the sensitivity of cells to H2O2 is greatly decreased in their ras-transformants. To understand the molecular basis of this sensitivity to H2O2, we attempted to identify H2O2-inducible cDNA clones from MC3T3 cells by differential screening of cDNA libraries, and one of such genes, named HIC-53, was isolated. The level of HIC-53 mRNA was moderately increased by H2O2 as well as by calcium ionophore or dexamethasone, but was not increased by the addition of serum, tumor promoting phorbol ester, or epidermal growth factor. Among mouse organs, HIC-53 mRNA levels were higher in the kidney and lung, but were almost undetectable in the brain, heart, bone, muscle or spleen. In MC3T3 cells transformed with v-Ki-ras, the HIC-53 mRNA level was markedly decreased, and effect of H2O2 was abolished. Although the biological function of HIC-53 is unknown at present, the predicted amino acid sequence exhibited some similarity with bovine cardiac Na+/Ca+ exchanger. The nucleotide sequence of HIC-53 cDNA showed no significant similarity with other known gene sequences.

Amino Acid Sequence↗
PubMed1995

Effects of the overexpression of the small heat shock protein, HSP27, on the sensitivity of human fibroblast cells exposed to oxidative stress.

The role of the human small heat shock protein (HSP27) in oxidative stress was examined using stable transformants of an immortalized human fibroblast cell line (KMST-6) isolated by transfection of HSP27 expression vectors. Several stable transformants that expressed high or low levels of HSP27 protein were obtained. Clones expressing high levels of HSP27 were more sensitive to growth inhibition by a low dose of hydrogen peroxide (0.1 mM) than those expressing low levels. Clones expressing high levels of HSP27 did not acquire obvious resistance to hyperthermy and cytotoxic agents, except for one (#13), in which resistance to cytotoxic agents was increased. The level of phosphorylated HSP27 in clones expressing high levels of this protein increased at 30 min and was sustained even 4 hours after exposing the cells to 0.1 mM of hydrogen peroxide. On the other hand, the levels in clones expressing low levels of HSP27 were reduced within 4 hours after exposure to hydrogen peroxide. Furthermore, overexpression of nonphosphorylatable mutant HSP27 did not affect sensitivity to oxidative stress. These results suggested that constitutively high expression of HSP27 in KMST-6 cells make them susceptible to oxidative stress resulting in growth arrest, and this mechanism could involve the phosphorylation of HSP27.

Cell Line, Transformed↗
PubMed1995

Activation of DNA synthesis and expression of the JE gene by catalase in mouse osteoblastic cells: possible involvement of hydrogen peroxide in negative growth regulation.

The addition of catalase isolated from bovine liver to the culture medium of quiescent mouse osteoblastic MC3T3-E1 cells decreased intracellular oxidized state, determined using fluorescent dye and laser-scanning confocal microscopy. The decrease in intracellular oxidized state evoked by catalase seemed to be involved in the arrest of growth, since catalase increased the incorporation of [3H]thymidine in these quiescent cells. On gel filtration of the catalase preparation, catalase activity and the stimulation of DNA synthesis coincided. Of the early response genes, JE, which is induced by competence factors, was induced by catalase in this cell line, whereas c-fos, c-jun, and KC mRNA levels were not affected. Catalase isolated from fungi and glutathione peroxidase+glutathione added to the culture medium also increased the steady-state level of JE mRNA. These results indicate that cells in the quiescent state produce hydrogen peroxide endogenously and that hydrogen peroxide acts as one of the mediators inhibiting DNA synthesis as well as the expression of JE, a growth factor-inducible gene.

3T3 Cells↗
PubMed1995

Increase in length of experimental skin flaps that survive with dibutyryl cyclic AMP.

One of the most important research topics in plastic surgery is the extension of the length of skin flaps that survive. We have investigated the increase in the length of skin flaps that can be achieved by giving dibutyryl cyclic AMP (DB-cAMP) to rabbits with experimental skin flaps and compared the results with those in animals not given DB-cAMP. Three variables, the arrival of DB-cAMP in the critical area of circulation of the flap (n = 6), changes in the blood flow in the flap (n = 10), and increase in the length of skin flap that survived (n = 30) were investigated by high performance liquid chromatography and laser Doppler flowmetry. DB-cAMP reached the critical area of circulation in the skin flap (dye distance), increased the blood flow within this area (mean (SEM) peak value 30 minutes after operation 1.24 (0.06) ml/min/kg compared with 1.06 (0.02) in control flaps), and extended the length of the flap that survived (mean (SEM) length seven days after operation 66.1 (3.0) mm compared with 60.8 (1.8) mm in the control group). We conclude that DB-cAMP improved the blood flow in skin flaps in rabbits with a consequent increase in the length of skin flap that survived.

Animals↗
PubMed1995

[Clinical statistics of 67 cadaveric renal transplantation at the Department of Urology, Kinki University School of Medicine].

A statistic survey was made on the patients undergoing cadaveric renal transplantation between June, 1980 and December, 1993. The total number of patients was sixty-seven. The number of patients per year increased annually. The graft survival rate was 73.8% at 1 year and 63.1% at 5 years. Some transition was seen in immunosuppressive regimens using cyclosporin.

Adolescent↗
PubMed1994

Characterization of the TGF beta 1-inducible hic-5 gene that encodes a putative novel zinc finger protein and its possible involvement in cellular senescence.

Transforming growth factor (TGF) beta 1 is a potent cytokine that inhibits the growth of several types of cells. Our earlier study suggested that the mouse osteoblastic cell line, MC3T3-E1, was sensitive to growth inhibition by TGF beta 1 and that this effect was partly mediated by H2O2. To identify the molecules that participate in the negative regulation of growth by these stimuli, we carried out differential screening of cDNA libraries and isolated a set of genes induced by TGF beta 1. Among the clones isolated, one originally named tsc-5 was found to be induced by H2O2 as well as TGF beta 1. Analysis of this cDNA renamed hic (hydrogen peroxide-inducible clone)-5 suggested that Hic-5 protein has four LIM motifs, each of which contained two (or one) putative zinc fingers. The expression of hic-5 mRNA was repressed in Ki-ras-transformed mouse fibroblasts and in several cell lines established from human tumor. On the other hand, its expression was augmented in the in vitro senescent process of human diploid fibroblasts. Among the mouse organs examined, hic-5 was highly expressed in the lung and spleen. Finally, a colony formation assay using an hic-5 expression vector driven by the cytomegalovirus promoter suggested that hic-5 overexpression had a cytostatic effect on cellular growth, depending upon the cell type. Although the relationship between hic-5 function and the signal transduction pathway of TGF beta 1 remains unresolved, these results implied that hic-5 has some role in the growth-inhibitory pathway associated with in vitro senescence, and that down-regulation of hic-5 contributes to tumorigenesis.

3T3 Cells↗
PubMed1994

Effect of radical scavengers on TNF alpha-mediated activation of the uPA in cultured cells.

Active oxygen, produced by cultured cells following stimulation with various growth factors, seems to be involved in signal transduction leading to cellular responses such as gene expression and growth modulation. In the present study, the intracellular oxidation state was measured in immortalized human endothelial cells (ECV304) after treatment with tumor necrosis factor (TNF) alpha, using a fluorescent dye and a laser-scanning confocal microscope. The intracellular oxidation state was increased 60 min after the addition of TNF alpha, and this increase was abolished by a radical scavenger, N-acetylcysteine (NAC), which is also a precursor of glutathione, and by pyrrolidine dithiocarbamate (PDTC). TNF alpha increased the steady state level of urokinase-type plasminogen activator (uPA), and NAC inhibited this increase at a dose that also inhibited the increase in the intracellular oxidation state. PDTC, on the other hand, did not affect the induction of the uPA gene by TNF alpha. These results suggest that intracellular glutathione level rather than the oxidation state is necessary for the induction of the uPA gene by TNF alpha.

Acetylcysteine↗
PubMed1994

Inhibition by N-acetyl-L-cysteine of interleukin-6 mRNA induction and activation of NF kappa B by tumor necrosis factor alpha in a mouse fibroblastic cell line, Balb/3T3.

Redox-based modulation plays a role in transcriptional control of gene expression. In the present study, we investigated the possible role of reactive oxygen species in the induction of interleukin-6 (IL-6) mRNA and in increases in NF kappa B binding activity by tumor necrosis factor (TNF) alpha using a mouse fibroblastic cell line, Balb/3T3. Expression of IL-6 mRNA is known to be dependent upon NF kappa B that binds to the 5'-flanking region of the IL-6 gene. We found that: (i) TNF alpha increased IL-6 mRNA levels and this increase was inhibited by N-acetyl-L-cysteine (NAC), a scavenger of reactive oxygen species. (ii) NF kappa B binding activity in this cell line was also increased by TNF alpha, and the increase was inhibited in the presence of NAC. (iii) The treatment of cells with low doses of hydrogen peroxide increased the NF kappa B binding activity. (iv) Expression of a reporter gene in which the chloramphenicol acetyltransferase (CAT) gene was under the control of NF kappa B binding sites was induced by hydrogen peroxide. These results suggest that the induction of IL-6 mRNA is regulated by a mechanism involving reactive oxygen species and that NF kappa B, whose activity is sensitive to the cellular redox state, plays an important role in this induction in a fibroblastic cell line, Balb/3T3, stimulated with TNF alpha.

3T3 Cells↗
PubMed1994

Induction of early response genes by hypergravity in cultured mouse osteoblastic cells (MC3T3-E1).

Hypergravity as low as 50g transiently stimulated cultured mouse osteoblastic cells (MC3T3-E1) to induce early response genes such as c-fos and egr-1, whereas expression of c-jun was marginally affected. The maximum induction of c-fos required more than 90g, but egr-1 induction became maximum below 50g. Staurosporin inhibited the induction of c-fos by hypergravity almost completely at a concentration of 0.1 microM, but it inhibited the induction of egr-1 only partially. In cells pretreated with 12-O-tetradecanoylphorbol 13-acetate, induction of c-fos by hypergravity was almost completely abolished, whereas that of egr-1 was not affected. Activity of protein kinase C seemed to be activated in cells centrifuged at 900g. These results indicate that hypergravity stimulates multiple signal transduction cascades that are connected with the expression of early response genes.

Animals↗
PubMed1994

Immaturity of the myenteric plexus is the aetiology of meconium ileus without mucoviscidosis: a histopathologic study.

The most common aetiology of meconium ileus is a deficiency in trypsin activity caused by cystic fibrosis. The pathogenesis of meconium ileus without mucoviscidosis is less well understood, although a number of causative factors have been suggested. The symptoms and clinical course of nine patients with meconium ileus without mucoviscidosis were reviewed, and the myenteric plexus of a surgical specimen of intestine was examined histologically and cytometrically. The nuclei of the intramural ganglion cells were much smaller than were seen in normal newborn infants. The nuclear areas resembled those seen in fetuses of 5-6 months gestational age, but the number of ganglion cells approached normal. This immaturity of the ganglia was observed both in the contracted distal ileum and dilated proximal ileum. Patients with an ileostomy passed solid faeces for about 1 to 2 months postoperatively, after which time the faeces became watery. The intramural ganglia were mature at the time of ileostomy closure. We conclude that immaturity of the myenteric plexus in the ileum and colon seems to be the main aetiologic factor in meconium ileus without mucoviscidosis.

Cytodiagnosis↗
PubMed1994

Extensive dissection at the porta hepatis for biliary atresia.

Dissection at the porta hepatis is the crucial step in surgery for biliary atresia. The authors describe their procedure for extensively dissecting the porta hepatis. The technique is based on an anatomic cast corrosion of the human liver, and the portal vein serves as the landmark for dissection. Lateral dissection at the hepatic hilus is critical to the procedure. On the right side, the anterior portal branches are dissected to the bifurcation of the S5 and S7 segmental portal branches, over the bifurcation of the anterior and posterior portal branches. On the left side, the liver parenchyma, which bridges the umbilical point, is sectioned, and the left portal branch is dissected to the umbilical point over the pars transversalis and taped. The fibrous mass that lies within the confines of the portal branches, including the dorsal aspect, is transected completely. This procedure was performed in 16 infants; the jaundice cleared in 93.7%.

Biliary Atresia↗
PubMed1994

Production of hydrogen peroxide by transforming growth factor-beta 1 and its involvement in induction of egr-1 in mouse osteoblastic cells.

TGF-beta 1 controls the expression of numerous genes, including early response and cellular matrix genes. However, the signal-transducing mechanism underlying this regulation of gene expression is not fully understood. In this study, we investigated whether redox regulation plays a role in the TGF-beta 1 signal transduction in the mouse osteoblastic cell line (MC3T3-E1). The overall intracellular oxidized state of the cells, when measured using 2',7'-dichlorofluorescin diacetate by laser-scanning confocal microscopy, was increased transiently after the addition of TGF-beta 1. This increase was abolished by the addition of oxygen radical scavengers such as catalase and N-acetylcysteine. In a variant cell line lacking the TGF-beta 1 receptor, the intracellular oxidized state was not modulated by treatment with TGF-beta 1. We then examined the expression of early growth response-1 (egr-1) gene, which is inducible by TGF-beta 1 and H2O2. Radical scavengers inhibited the induction of egr-1 by TGF-beta 1, but not that by 12-O-tetradecanoylphorbol-13 acetate. A nuclear run-on assay indicated that this inhibition was at the transcriptional level. From transient expression experiments using chloramphenicol acetyltransferase gene linked to serially deleted egr-1 gene 5'-upstream region, the CArG element in the 5' flanking region of egr-1 was identified as an essential sequence in the transcriptional activation for both TGF-beta 1 and H2O2 stimulation. These findings suggest that H2O2 acts as a mediator for the TGF-beta 1-induced transcription of egr-1 gene.

3T3 Cells↗
PubMed1993

Cloning from a mouse osteoblastic cell line of a set of transforming-growth-factor-beta 1-regulated genes, one of which seems to encode a follistatin-related polypeptide.

Transforming growth factor(TGF)beta 1 is a potent inhibitor of growth in mouse osteoblastic MC3T3-E1 cells. To isolate genes that are induced by TGF beta 1, the differential screening method was adopted using a cDNA library constructed from cells treated with TGF beta 1 for 4 h. Six independent cDNA clones were isolated (TGF beta-stimulated clone, TSC-5, TSC-36, TSC-115, TSC-128, TSC-160 and TSC-161), the expression of which was increased by TGF beta 1-treatment with maximal expression at 6-10 h. The steady-state levels of TSC-36, TSC-128 and TSC-160 increased almost tenfold, whereas those of TSC-5, TSC-115 and TSC-161 were elevated at most threefold. From partial nucleotide sequences, TSC-160 was found to be identical to rrg (ras-recision gene, lysyl oxidase), and TSC-115 had 80% similarity with tropomyosin cDNA, whereas other genes seemed novel. Expression of TSC-36 and TSC-160 was dramatically decreased in v-Ki-ras-transformed MC3T3 cells or in transformed NIH 3T3 cells (DT), and was recovered to normal levels in a flat revertant (C11). A nearly full-length copy of TSC-36 cDNA was isolated, and an open reading frame indicated that it encodes a protein of 35 kDa. An antiserum was raised against the C-terminal peptide predicted from the nucleotide sequence, and a polypeptide with an approximate molecular mass of 38 kDa was detected in cultured medium of MC3T3-E1 cells. The amino acid sequence of TSC-36 protein was found to have some similarity with follistatin, an activin-binding protein, and a limited similarity with the secreted protein rich in cysteine (SPARC).

3T3 Cells↗
PubMed1993

Common target for 12-O-tetradecanoylphorbol-13-acetate and ras in the transcriptional enhancer of the growth factor-inducible JE gene.

JE gene expression in the mouse osteoblast cell line MC3T3-E1 is activated transiently by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). In ras-transformed MC3T3-E1 cells the JE gene is constitutively expressed at a high level, whereas in their raf-transformed counterparts it is not constitutively expressed or inducible by TPA. Using these cells, we investigated a specific sequence recognized by nuclear factors in the 5'-upstream region of the rat JE gene. By gel-mobility shift assays, we determined that the amount of nuclear factors that bind to the region -130 to -96 bp upstream from the cap site of the rat JE gene (JE-1 probe) increased after TPA treatment of MC3T3-E1 cells. However, in the ras transformants it was elevated constitutively, and in the raf transformants it was not detectable. Both unlabeled JE-1 probe and a probe containing a TPA-responsive element (TGACTCA) competed with the binding of these nuclear factors. Preincubation of the nuclear extracts with fos- or jun-specific antibodies interfered with the binding of the factors to the JE-1 probe. The essential sequence in the JE-1 element for the binding of nuclear factors was found to be TTACTCA. c-fos and c-jun proteins synthesized in vitro could bind to the DNA fragment containing this sequence, but the binding was weaker than to the TPA-responsive element (TGACTCA). These results suggest that the sequence TTACTCA in the JE-1 element is a common target of TPA and ras and that protein complexes containing fos- and jun-related proteins recognize the sequence.

Animals↗
PubMed1993

Clinical, laboratory and prognostic features of congenital large intestinal motor dysfunction (pseudo-Hirschsprung's disease).

One hundred and forty-eight cases of congenital large intestinal motor dysfunction (pseudo-Hirschsprung's disease) were reported by members of the Japanese Society of Pediatric Surgeons during the past 20 years. The disorder was defined as a congenital, non-mechanical obstruction of the intestine with the presence of intramural ganglia in the terminal rectum. Intramural ganglia were abnormal in 77 cases, normal in 42, and could not be determined in 29. Of those with abnormal intramural ganglia, 54 had immature ganglia or hypoganglionosis (oligoganglionosis), 15 had neuronal intestinal dysplasia, and eight had a segmental anomaly. Of those with a normal myenteric plexus, 22 had chronic and twelve had suspected idiopathic intestinal pseudo-obstruction syndrome; eight had megacystis-microcolon-intestinal hypoperistalsis syndrome. While cases with both hypoganglionosis and normal intramural ganglia had normal acetylcholine esterase activity, a significantly greater number of patients with hypoganglionosis lacked normal rectoanal reflexes. Patients with hypoganglionosis, chronic idiopathic intestinal pseudo-obstruction syndrome, and megalocystis-microcolon-intestinal hypoperistalsis syndrome had poor prognoses with an overall mortality of 36.9%. These findings indicate that congenital large intestinal motor dysfunction remains a serious disease of childhood.

Age of Onset↗
PubMed1993

Outcome of 21 patients with biliary atresia living more than 10 years.

The outcome of 21 survivors (8 males, 13 females; age range, 11 to 29 years) followed for > 10 years after surgery for biliary atresia are discussed. Of the 21 patients, 18 with type III disease had hepatic portoenterostomy (Kasai operation), and 3 with type I disease had hepaticoenterostomy. Twenty patients are alive leading almost normal lives; however, 13 (61.9%) patients did have a history of complications, including hemorrhage from esophageal varices in 10, from a gastric ulcer or erosion in 3, and from a duodenal ulcer in 2; biliary reobstruction in 3; and multiple pulmonary arteriovenous fistulae in 2. Of those with complications, 7 required surgery. One died suddenly at the age of 19 years of a bleeding gastric ulcer. Liver function is normal in 9 (45.0%) of the 20 alive patients, 2 (10.0%) have slight hepatic dysfunction, and 9 (45.0%) have mild-to-moderate hepatic dysfunction. Liver function is almost normal in 3 patients who had a history of variceal hemorrhage.

Adolescent↗