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T Noguchi

Publications and source records attributed to T Noguchi.

At least 271 records · Page 15Linked to original sources
PubMed1998

Integrin-mediated tyrosine phosphorylation of SHPS-1 and its association with SHP-2. Roles of Fak and Src family kinases.

SHPS-1 is a receptor-like glycoprotein that undergoes tyrosine phosphorylation and binds SHP-2, an Src homology 2 domain containing protein tyrosine phosphatase, in response to various mitogens. Cell adhesion to extracellular matrix proteins such as fibronectin and laminin also induced the tyrosine phosphorylation of SHPS-1 and its association with SHP-2. These responses were markedly reduced in cells overexpressing the Csk kinase or in cells that lack focal adhesion kinase or the Src family kinases Src or Fyn. However, unlike Src, focal adhesion kinase did not catalyze phosphorylation of the cytoplasmic domain of SHPS-1 in vitro. Overexpression of a catalytically inactive SHP-2 markedly inhibited activation of mitogen-activated protein (MAP) kinase in response to fibronectin stimulation without affecting the extent of tyrosine phosphorylation of focal adhesion kinase or its interaction with the docking protein Grb2. Overexpression of wild-type SHPS-1 did not enhance fibronectin-induced activation of MAP kinase. These results indicate that the binding of integrins to the extracellular matrix induces tyrosine phosphorylation of SHPS-1 and its association with SHP-2, and that such phosphorylation of SHPS-1 requires both focal adhesion kinase and an Src family kinase. In addition to its role in receptor tyrosine kinase-mediated MAP kinase activation, SHP-2 may play an important role, partly through its interaction with SHPS-1, in the activation of MAP kinase in response to the engagement of integrins by the extracellular matrix.

Animals↗
PubMed1998

Allosteric regulation of pyruvate kinase M2 isozyme involves a cysteine residue in the intersubunit contact.

Pyruvate kinase M2 isozyme mutants with amino acid substitutions in the subunit interface were prepared and characterized. The substitutions were made in the allosteric M2 isozyme by the corresponding residues of the nonallosteric M1 isozyme to identify the residue involved in the allosteric effects. The replacement of Cys-423 by Leu led to substantial loss of both homotropic and heterotropic allosteric effects while the substitutions at Phe-389, Arg-398, Ala-401, Pro-402, Thr-408, and Ile-427 did not. The altered kinetic properties of the Cys-423-substituted mutant resulted from the shift of the allosteric transition toward the active R-state since the mutant exhibits the allosteric properties in the presence of an allosteric inhibitor, L-phenylalanine. The inverse correlation between the hydrophobicity of residue 423 and the extent of stabilization of the R-state was found by analysis of mutants with un-ionizable amino acids at position 423. Furthermore, the modification of Cys-423 with methyl methanethiosulfonate led to a shift of the allosteric transition toward the R-state, probably the result of increased hydrophobicity of the residue. These results suggest that Cys-423 is involved in the allosteric regulation of the enzyme through hydrophobic interactions.

Allosteric Regulation↗
PubMed1998

Effects of cyclosporine and FK506 on in vitro high shear-induced platelet reactivity in rat and human non-anticoagulated blood.

BACKGROUND: The immunosuppressants cyclosporine and FK506 have been used successfully in clinical transplantation, but both agents have various side effects. We have previously found that cyclosporine is prothrombotic and that FK506 is antithrombotic in an in vivo system. The aim of the present study was to assess the effects of these agents on platelet reactivity and coagulation using an in vitro shear-induced hemostatic platelet plug-forming instrument, the hemostatometer. METHODS: A purpose-built hemostatometer was constructed in our laboratory. The effects of cyclosporine and FK506 on platelet reactivity and coagulation were assessed under high shear stress using non-anticoagulated rat and human blood. RESULTS: FK506 significantly inhibited both platelet reactivity and coagulation. Cyclosporine also significantly inhibited coagulation, but a proaggregatory effect was observed at a final blood concentration of 0.05 mg/ml. CONCLUSIONS: The present in vitro results support our previous in vivo findings regarding the prothrombotic and antithrombotic effects of cyclosporine and FK506, respectively.

Adult↗
PubMed1998

Expression of Hex mRNA in early murine postimplantation embryo development.

The onset of Hex expression and its role in early murine development was analyzed using in situ hybridization. Hex mRNA was first detected in the chorion of the ectoplacental cavity and weakly at the visceral endoderm of the future yolk sac at embryonic age (E) 7.5. Expression in embryonic tissues was detected exclusively in the hepatic anlage and thyroid primordium at E 9.5. At E 12.5 and E 15.5, Hex expression persisted in the fetal liver and thyroid, and was also detected in the fetal lung. These results suggest that Hex has its role in differentiation and/or organogenesis of several embryonic tissues.

Animals↗
PubMed1998

Mutations at BRCA1: the medullary breast carcinoma revisited.

BRCA1-associated breast cancers (BRCA1-BCs) frequently harbor a high histoprognostic grade, p53 alterations, and estrogen receptor negativity. Although these parameters predict a poor outlook, the overall survival in BRCA1-BCs is equivalent to or even better than that in sporadic cases. These features are reminiscent of what is observed for breast carcinoma of the medullary type, a high-grade tumor with a particular favorable course. To explore a possible relationship between this phenotype and BRCA1 mutations, we first compared 32 BRCA1-BCs and 200 consecutive cases of breast cancer without familial history for the prevalence of typical medullary breast carcinoma (TMC) using the criteria given by Ridolfi et al. [R. Ridolfi et al, Cancer (Phila.), 40: 1365-1385, 1977]. Second, we searched for BRCA1 mutations in a set of 18 cases of TMC, using denaturing gradient gel electrophoresis and Cleavase fragment length polymorphism scanning. Six of 32 (19%) BRCA1-BCs were of the TMC type, compared to 0 of 200 controls (P < 0.0001). Among the 18 TMCs, 2 BRCA1 nonsense mutations were found. This corresponds to almost 7 times the contribution of BRCA1 mutations in the general population. Two additional missense mutations were identified. Together, these results suggest that, although TMC and BRCA1-BCs are not strictly coincidental, an important connection between the two populations does exist.

BRCA1 Protein↗
PubMed1998

Roles of the complex formation of SHPS-1 with SHP-2 in insulin-stimulated mitogen-activated protein kinase activation.

SHPS-1 is a receptor-like protein that undergoes tyrosine phosphorylation and binds SHP-2, an SH2 domain-containing protein tyrosine phosphatase, in response to insulin and other mitogens. The overexpression of wild-type SHPS-1, but not of a mutant SHPS-1 in which all four tyrosine residues in its cytoplasmic region were mutated to phenylalanine, markedly enhanced insulin-induced activation of mitogen-activated protein kinase in Chinese hamster ovary cells that overexpress the human insulin receptor. Mutation of each tyrosine residue individually revealed that the major sites of tyrosine phosphorylation of SHPS-1 in response to insulin are Tyr449 and Tyr473. In addition, mutation of either Tyr449 or Tyr473 abolished the insulin-induced tyrosine phosphorylation of SHPS-1 and its association with SHP-2. Surface plasmon resonance analysis showed that glutathione S-transferase fusion proteins containing the NH2-terminal or COOH-terminal SH2 domains of SHP-2 bound preferentially to phosphotyrosyl peptides corresponding to the sequences surrounding Tyr449 or Tyr473, respectively, of SHPS-1. Furthermore, phosphotyrosyl peptides containing Tyr449 or Tyr473 were effective substrates for the phosphatase activity of recombinant SHP-2 in vitro. Together, these results suggest that insulin may induce phosphorylation of SHPS-1 at Tyr449 and Tyr473, to which SHP-2 then binds through its NH2-terminal and COOH-terminal SH2 domains, respectively. SHPS-1 may play a crucial role both in the recruitment of SHP-2 from the cytosol to a site near the plasma membrane and in increasing its catalytic activity, thereby positively regulating the RAS-mitogen-activated protein kinase signaling cascade in response to insulin.

Amino Acid Sequence↗
PubMed1998

[Prognostic factors in stomach carcinoma].

The classical prognostic factors of the pTNM system are still most valid. Nevertheless, vascular invasion as well as the molecular marker E-cadherin proved to be independent new prognostic factors responsible for a significant shift in patient survival. Thus, in pTNM-stage II patients, a highly significant drop in survival is observed when patients showing no E-cadherin expression and at the same time vascular invasion are compared with E-cadherin-positive patients who do not show vascular invasion (Fig. 11). These conspicious shifts in survival underline the necessity to continue our search for new molecular and non-molecular markers which in future may help us to predict the outcome of gastric cancer patients more precisely and more individually.

Antigens, CD↗
PubMed1998

Loss of heterozygosity at loci from chromosome arm 22Q in human sporadic breast carcinomas.

Forty-four breast carcinomas were studied for loss of heterozygosity (LOH) at 25 microsatellite markers distributed almost evenly along chromosome arm 22q. LOH at at least one marker were observed in 66% tumors, while 6 regions of consistent LOH were identified. The size of each region ranged between 3 and 6 cM, and the distance between each region was estimated to be 8 to 12 cM. Even if not all these regions contain a bona fide tumor-suppressor gene, it is possible that several loci from chromosome arm 22q may be involved in breast carcinogenesis.

Breast Neoplasms↗
PubMed1998

Response of chondrocyte subpopulations cultured within unloaded and loaded agarose.

Although it is well known that the metabolism of chondrocytes can be altered by the application of mechanical strain, it is unclear whether chondrocytes from the superficial and deep zones of cartilage respond in a similar manner. In this study, chondrocytes from the uppermost 15-20% (superficial cells) and the remaining tissue (deep cells) from bovine articular cartilage were isolated separately and cultured in agarose constructs. Cell deformation on application of a 15% static compressive strain was identical for both subpopulations after 24 and 72 hours in culture. The constructs were incubated under static and dynamic (0.3, 1, and 3 Hz) strains of 15% amplitude. Glycosaminoglycan synthesis by deep cells was unaffected by static strain or 3 Hz dynamic strain, whereas 0.3 Hz produced a significant reduction and 1 Hz induced a highly significant 50% stimulation of glycosaminoglycan synthesis (p < 0.001). Superficial cells exhibited a general inhibition of glycosaminoglycan synthesis. By contrast, proliferation of superficial cells was stimulated by dynamic strain whereas deep cells were not influenced. It has been suggested previously that mechanotransduction-induced controls of glycosaminoglycan synthesis and proliferation in chondrocytes embedded in agarose are uncoupled. Data presented in this study demonstrate that the two processes do, in fact, occur in different subpopulations of chondrocytes within the full-depth cell isolate.

Animals↗
PubMed1998

Isolation and characterization of transposon-induced mutants of Porphyromonas gingivalis deficient in fimbriation.

Fimbriae are considered to be an important virulence factor of Porphyromonas gingivalis. In order to identify genes essential for fimbriation, other than fimA which encodes the major subunit protein of fimbriae, transposon mutagenesis and immunological screening techniques were used to isolate fimbria-deficient mutants. R751::*Omega4, a suicide vector that carries Tn4351, was transferred from Escherichia coli to P. gingivalis by conjugation. Twenty-two independent fimbria-deficient mutants were identified among the resulting transformants. Southern hybridization analysis with pBlue 4351, a transposon-specific probe, and R751 indicated that 45% of the mutants resulted from single transposon insertions and that the remaining 55% of the mutants resulted from cointegration of R751 sequences. Southern hybridization analysis with pUCBg12.1, a probe for the fimA region, indicated that nine of the mutants contained insertions within the 2.5 kb SacI DNA fragment of P. gingivalis that contains fimA, ORF1 (which encodes a 15 kDa protein), and the C-terminal portion of ORF5 (which encodes a 63 kDa protein). Polymerase chain reaction (PCR) analysis and further Southern hybridization analysis indicated that the insertion site(s) for all nine of these mutants was within the fimA gene. Southern hybridization analysis also indicated that the remaining thirteen mutants contained insertions somewhere outside the 10 kb fimA region. Analysis by pulsed field gel electrophoresis (PFGE) revealed that insertions for most of the thirteen mutants mapped to a 300 kb NotI fragment and are located at least approximately 200 kb away from fimA. These results identify genetic loci other than fimA, that are required for fimbriation of P. gingivalis. Future cloning and characterization of these genetic loci should be straightforward since they are now marked by antibiotic resistance genes carried by the transposon.

Bacterial Proteins↗
PubMed1998

Evaluation of self-similar features in time series of serum growth hormone and prolactin levels by fractal analysis: effect of delayed sleep and complexity of diurnal variation.

We assayed the diurnal concentrations of growth hormone (GH) and prolactin (PRL) in 6 healthy male volunteers to evaluate the self-similar features in the time series of each hormone on the basis of fractal theory and to determine the fractal dimension as an index of the complexity of the diurnal variation. In addition, we assessed the effects of a 6-hour delay in the sleep period on the complexity of the diurnal variation of these hormones. There was a statistically significant fractal feature in the serum levels of GH both under the nocturnal-sleep and delayed-sleep conditions in all subjects. The time series of the serum PRL concentrations also showed a statistically significant fractal feature under the nocturnal-sleep and delayed-sleep conditions in all subjects. The fractal dimensions of the patterns of the GH or PRL levels were 1.879 and 1.929 or 1.754 and 1.785 under the nocturnal-sleep and delayed-sleep conditions, respectively. Two-way ANOVA revealed no significant difference in the fractal dimension between the two sleep conditions but did reveal a significant difference between the fractal dimensions of the GH and PRL levels. These results showed (1) that delayed sleep had no significant effect on the complexity of the diurnal pattern of these hormones, and (2) that the diurnal pattern of the GH levels was more complex than that of the PRL levels.

Adult↗
PubMed1998

The surgical treatment of lung lacerations and major bronchial disruptions caused by blunt thoracic trauma.

The records of 16 patients who suffered blunt thoracic trauma, causing lung lacerations in 13, bronchial disruptions in 2, and lung laceration with bronchial disruption in 1, were reviewed to investigate the correlations between clinical factors and prognosis. The causes of these injuries included 14 traffic accidents and 2 construction-related accidents, and the indications for surgery were massive bleeding in 12 patients, massive air leakage in 2, both in 1, and lung abscess in 1. Of the 16 patients, 11 (68.8%) underwent thoracotomy less than 4 h after admission, 3 (18.8%) underwent thoracotomy 4 to 24 h after admission, and 2 (12.5%) underwent thoracotomy 24 h or later after admission. The operative techniques included 1 pneumonectomy, 5 lobectomies, 2 bronchoplasties, and 8 minor repairs. The mortality rate was 43.7%, which included six early deaths occurring within 72 h of the trauma, and one late death. While major bronchial disruption is usually associated with a good prognosis, univariate and multivariate analyses demonstrated that intrapleural bleeding of 300 ml/h or more from time of trauma to chest tube drainage was significantly correlated with a poor prognosis. Moreover, an injury severity score (ISS) of 36 or more showed a trend toward a correlation with poor prognosis in patients with lung lacerations. Prompt thoracotomy will decrease mortality rate of patients suffering lung lacerations resulting in intrapleural bleeding of more than 300 ml/h.

Adolescent↗
PubMed1998

Leptin changes Ca2+/calmodulin-dependent response and up-regulates the gene expression of calcineurin in rat hypothalamus.

To understand the leptin's action on hypothalamic arcuate nucleus (ARC), Ca2+/calmodulin (CaM)-dependent response in leptin-treated rat ARC was investigated. The in vitro phosphorylation of ARC extracts from leptin-treated rats did not respond to CaM. Although CaM-dependent protein kinase II activity was not affected by leptin, the gene expression of calcineurin, CaM-dependent phosphatase, increased in leptin-treated rats.

Animals↗
PubMed1998

Brassinosteroids in Arabidopsis thaliana.

From the seeds and siliques of Arabidopsis thaliana, six brassinosteroids, brassinolide, castasterone, typhasterol, 6-deoxocastasterone, 6-deoxotyphasterol and 6-deoxoteasterone, were identified by GC-mass spectrometry or GC-selected ion monitoring. As the occurrence of castasterone, typhasterol, 6-deoxocastasterone and 6-deoxotyphasterol in the shoots of A. thaliana has already been reported, this study provides evidence for the occurrence of the above four brassinosteroids in different organs, seeds and siliques, and the first evidence for the occurrence of brassinolide and 6-deoxoteasterone in A. thaliana. All brassinosteroids identified in this study belong to important components of both the early and late C-6 oxidation pathways, which were established in the cultured cells of Catharanthus roseus. This suggests that both pathways are operating in A. thaliana to produce the most biologically active brassinosteroid, brassinolide, which is responsible for growth and development of the plant.

Arabidopsis↗
PubMed1998

Phytochrome-mediated light signals are transduced to nucleoside diphosphate kinase in Pisum sativum L. cv. Alaska.

To clarify the molecular mechanism for the transduction of light signals in plants, we have established an in vitro system that uses crude membrane and soluble fractions of stem sections of etiolated Pisum sativum L. cv. Alaska after irradiation by red light, or sequential application of red and far-red light to the stem section. In a previous report (T. Hamada et al., J. Photochem. Photobiol. B: Biol. 33 (1996) 143-151) the labelling of proteins in membrane fraction by [gamma-32P] ATP at 0 degree C for 15 s and subsequent separation of proteins by two-dimensional electrophoresis allowed unambiguous identification of a heavily phosphorylated protein spot at 18 kDa (p18). In the present study we have confirmed the former results in the membrane fraction, and obtained the result that an increase in the phosphorylation of p18 by red-light irradiation is observed in the soluble fraction. Further, we have provided evidence that the p18 in the soluble fraction is purified and identified as nucleoside diphosphate (NDP) kinase by Western blotting, immuno-precipitation, amino acid sequencing and cDNA analysis. Purified p18 shows autophosphorylation activity and strong phosphorylating activity against myelin basic protein (MBP), a substrate of MAP (mitogen activated protein) kinase. The results show that phytochrome-mediated light signals are transduced to NDP kinase, which may elicit signals by providing high concentrations of, for example, GTP from GDT and ATP, by the autophosphorylation and by the protein kinase activity similar to MAP kinase.

Alaska↗
PubMed1998

Epidemiology of major trauma and trauma deaths in Los Angeles County.

BACKGROUND: Our objective was to study population-based trauma-related injuries and deaths in the county of Los Angeles and to identify trends and progress towards meeting the "Year 2000 National Health Objectives." STUDY DESIGN: We did a retrospective study for the year 1996. Data were obtained from the Trauma Registry of the Emergency Medical Services of the Department of Health Services, and the Coroner's Department of the County of Los Angeles. Traumatic injuries and deaths per 100,000 of the population were calculated according to mechanism, race, age, and gender. RESULTS: During 1996, there were 12,136 major trauma admissions in the 13 trauma centers in Los Angeles County. Another 1,929 victims died at the scene or were certified dead at nontrauma centers and were taken to the Coroner's Department (total 14,065 victims). The overall major injury rate was 151.0 per 100,000 population and the death rate was 30.9 per 100,000. The trauma death rate per 100,000 population was 56.4 for African-Americans, 33.5 for Hispanics, 26.3 for Caucasians, and 11.6 for Asians. Homicides were the leading cause of traumatic deaths (45.3%) followed by traffic accidents (31.9% of deaths). Firearms were responsible for 3,899 major injuries or deaths (41.7 per 100,000 population). The overall homicide rate per 100,000 population was 14.0, with a much higher rate for African-Americans (40.4 per 100,000) and Hispanics (18.7 per 100,000) than Caucasians (4.0 per 100,000) or Asians (3.4 per 100,000). African-American males were at very high risk for homicide (73.3 per 100,000), and in the age group 15 to 34 years, this problem reaches epidemic proportions (164.2 per 100,000). Traffic accidents accounted for 69.0 major injuries and 9.6 deaths per 100,000 people. Males were at significantly higher risk of dying in traffic accidents than females. People over 60 years of age were at significantly higher risk of traffic-accident death than younger people, for both passenger and pedestrian groups (p < 0.01). Firearm-related suicides were responsible for 4.6 deaths per 100,000 population. Caucasian males over 65 years were at much higher risk of suicide by penetrating trauma (29.5 per 100,000) than were Hispanics (6.3 per 100,000), Asians (5.4 per 100,000), or African-Americans (no deaths) in the same gender and age group. CONCLUSIONS: Trauma remains a major health problem in the county of Los Angeles. Despite the significant reduction of intentional trauma in 1996, it still exceeds national figures and is much higher than the targeted "Year 2000 National Health Objectives." Aggressive prevention strategies need to focus on the population groups at excessive risks of injury by assault, traffic accidents, and suicides.

Accidents, Traffic↗
PubMed1998

Selection system for genes encoding nuclear-targeted proteins.

Nuclear proteins have essential roles in cell proliferation and differentiation. We have developed a yeast selection system-the nuclear transportation trap (NTT)-to identify genes encoding nuclear transport signals. Both unknown and previously identified nuclear localization signals were identified from a human fetal brain cDNA library. The majority (75%) of the unknown proteins examined were exclusively localized to the nucleus in COS-7 cells. We propose that NTT is an efficient method for isolating cDNAs that encode nuclear targeted proteins that can be applied to the retrieval of novel nuclear proteins and to annotate gene function.

Amino Acid Sequence↗