Biomedical subjects

T Uno

Publications and source records attributed to T Uno.

At least 73 records · Page 4Linked to original sources

PubMed1998

Relative involvement of Shc tyrosine 239/240 and tyrosine 317 on insulin induced mitogenic signaling in rat1 fibroblasts expressing insulin receptors.

Shc is phosphorylated on Tyr-239/240 and/or Tyr-317, which serves as a docking site for Grb2. To clarify the relative involvement of Shc Tyr-239/240 and Tyr-317 in insulin-induced mitogenesis, we generated expression vectors for Y317F (1F)-Shc, Y239/240F (2F)-Shc, and Y239/240/317F (3F)-Shc, and stably transfected them into Rat1 fibroblasts expressing insulin receptors (HIRc). Insulin-induced Shc phosphorylation and subsequent association with Grb2 was enhanced in wild-type (WT)-Shc cell. In contrast, insulin-stimulated Shc phosphorylation and Shc.Grb2 association were significantly decreased in 1F-Shc and 3F-Shc cells, while these were only slightly affected and almost comparable in 2F cells compared with those in parental HIRc cells. The kinetics of MAP kinase activation closely paralleled the kinetics of Shc phosphorylation and Shc.Grb2 association. Thus, insulin stimulation of MAP kinase activation occurred more rapidly in WT-Shc cells, and the activation was delayed in 1F-Shc and 3F-Shc cells, while it was comparable in 2F-Shc cells compared with that in HIRc cells. Furthermore, WT-Shc cells displayed enhanced sensitivity to insulin stimulation of thymidine incorporation. Importantly, the sensitivity was significantly decreased in 1F-Shc and 3F-Shc cells, while it was almost comparable in 2F-Shc cells compared with that in HIRc cells. These results indicate that Shc Tyr-317 is more predominant insulin-induced phosphorylation site than Tyr-239/240 for coupling with Grb2 leading to MAP kinase activation and mitogenesis in Rat1 fibroblasts.

Adaptor Proteins, Signal Transducing↗

PubMed1998

Preoperative Diagnosis of Breast Diseases by Dynamic MR Mammography:Cut-off Point Establishment for Signal Intensity Ratio.

We have demonstrated that the dynamic study for breast lesions by magnetic resonance imaging(MRI)can differentiate benign from malignant lesions objectively.The cases were 57 histopathologically appraised breast lesions, including 20 cases of breast cancer, 28 cases of mastopathy, 8 cases of fibroadenoma and 1 caseof intraductal papillomatosis. We plotted time-signal intensity ratio curves and then determined 95% confidence intervals, plotting the signal intensity ratio for both breast cancer and mastopathy every 30 seconds during dynamic magnetic resonance mammography(MRM)after gadolinium-diethylenetriamine pentaacetic acid(Gd-DTPA)administration, and further established cut-off points to differentiate between them. We then tried of estimate objectively the benign-malignant differentiation to breast lesions by confirming their signal intensity ratio to be more or less than the cut-off points. We advocate this procedure, and call it the " dynamic ratio method. " As a result, we found highly significant differences between breast cancer and mastopathy at 30 and 60 seconds after Gd-DTPA administration(P < 0.0001). We also confirmed that the cut-off point for the dynamic ratio method was equivalent to 1.4 and 1.8 times the precontrast signal intensity value at30 and 60 seconds after administration of Gd-DTPA respectively. By performing this dynamic ratio method preoperatively we can assess objectively not only the malignancy of breast lesions, but also neighboring infiltration, extending intraductal component, and lymph node metastasis. Furthermore, the dynamic ratio method provides detailed information for selecting the appropriate region for breast conserving surgery preoperatively, and can be expected to reduce unnecessary biopsies of benign cases. The dynamic ratio method had a sensitivity of 95.0%, a specificity of 81.1% and a positive predictive value of 73.1%. Also, for detectinginvasive ductal carcinoma, the sensitivity of the dynamic ratio method was 100.0%.

Journal Article↗

PubMed1998

A combinatorial approach to the discovery of efficient cationic peptoid reagents for gene delivery.

A family of N-substituted glycine oligomers (peptoids) of defined length and sequence are shown to condense plasmid DNA into small particles, protect it from nuclease degradation, and efficiently mediate the transfection of several cell lines. The oligomers were discovered by screening a combinatorial library of cationic peptoids that varied in length, density of charge, side-chain shape, and hydrophobicity. Transfection activity and peptoid-DNA complex formation are shown to be highly dependent on the peptoid structure. The most active peptoid is a 36-mer that contains 12 cationic aminoethyl side chains. This molecule can be synthesized efficiently from readily available building blocks. The peptoid condenses plasmid DNA into uniform particles 50-100 nm in diameter and mediates the transfection of a number of cell lines with efficiencies greater than or comparable to DMRIE-C, Lipofectin, and Lipofectamine. Unlike many cationic lipids, peptoids are capable of working in the presence of serum.

3T3 Cells↗

PubMed1998

Substitutions of conserved aromatic amino acid residues in subunit I perturb the metal centers of the Escherichia coli bo-type ubiquinol oxidase.

Cytochrome bo is a four-subunit quinol oxidase in the aerobic respiratory chain of Escherichia coli and functions as a redox-coupled proton pump. Subunit I binds all the redox metal centers, low-spin heme b, high-spin heme o, and CuB, whose axial ligands have been identified to be six invariant histidines. This work explored the possible roles of the aromatic amino acid residues conserved in the putative transmembrane helices (or at the boundary of the membrane) of subunit I. Sixteen aromatic amino acid residues were individually substituted by Leu, except for Tyr61 and Trp282 by Phe and Phe415 by Trp. Leu substitutions of Trp280 and Tyr288 in helix VI, Trp331 in loop VII-VIII, and Phe348 in helix VIII reduced the catalytic activity, whereas all other mutations did not affect the in vivo activity. Spectroscopic analyses of the purified mutant enzymes revealed that the defects were attributable to perturbations of the binuclear center. On the basis of these findings and recent crystallographic studies on cytochrome c oxidases, we discuss the possible roles of the conserved aromatic amino acid residues in subunit I of the heme-copper terminal oxidases.

Amino Acid Sequence↗

PubMed1998

High dose rate brachytherapy for carcinoma of the cervix: risk factors for late rectal complications.

PURPOSE: To determine the incidence of late rectal complications in patients treated with high dose rate brachytherapy for FIGO Stage IIB, IIIB carcinoma of the uterine cervix, and to evaluate the treatment factors associated with an increased probability of treatment complications. METHODS AND MATERIALS: Records of 100 patients with FIGO IIB and IIIB cervical carcinoma treated with definitive irradiation using high dose rate intracavitary brachytherapy (HDR-ICR) between 1977 and 1994 were retrospectively reviewed. For each HDR-ICR session, 6-Gy isodose volume was reconstructed retrospectively and the relationship between probability of late rectal complications and several treatment factors, including a specific point dose and parameters representing isodose volume, were examined. Statistical analyses were performed to determine the treatment factors predictive of late rectal complications. RESULTS: Of patients treated for both stages, 33% and 38% had experienced moderate to severe (Grade 2-4) complications at 3 and 5 years, respectively. Mean value of depth (D) of 6-Gy isodose volume in HDR-ICR in patients with and without complication were 51 mm and 46 mm, respectively (p = 0.0070). A significant difference was noted in complication rate between patients with D > 51 mm and D < or = 51 mm (p = 0.0023). Cumulative Point S (2 cm dorsal from the midpoint of the ovoid sources) dose (p = 0.044), and single or total point S dose by HDR-ICR (p = 0.019, each) were significantly higher in patients who developed complication, whereas these factors did not significantly affect the probability of pelvic control. Multivariate analysis revealed that D was the independent predictor for the endpoint of actuarial complication rate (p = 0.047). No significant difference was noted in the product of L, D, and W value (L x D x W) between patients with less than Grade 2 rectal complication and those with Grade 2-4. CONCLUSION: Depth of 6-Gy isodose volume determined three dimensionally (3D) has the predictive value of late rectal complications. This suggests that the shape of the high dose area in HDR-ICR influences the incidence of late rectal complications regardless of its volume.

Adult↗

PubMed1998

Squamous cell carcinoma of the maxillary sinus treated with radiation therapy and conservative surgery.

BACKGROUND: For patients with squamous cell carcinoma of the maxillary sinus, combined modality treatment is usually employed, involving radiation and en bloc radical surgery. In this study, local control was analyzed retrospectively in patients who underwent less aggressive piecemeal surgery. METHODS: Of the 37 patients irradiated between 1973 and 1992, 62% were classified as having T4 lesions. Thirty-two patients underwent surgery and radiation therapy; conventional fractionation radiation therapy was used in most cases. Thirty of these patients underwent piecemeal debulking of their tumors and simultaneous radiation therapy. RESULTS: Local recurrence free survival at 5 years was 59%, and orbital exenteration was performed on only 1 patient. T classification, the number of operations, and the presence of macroscopic residual disease each had a statistically significant impact on local recurrence. For the patients with macroscopic residual disease, more than 58 gray administered in conventional fractionation appeared to be necessary to improve local control. CONCLUSIONS: Combined with radiation therapy, conservative surgery with piecemeal debulking was an effective method of treatment for the patients in this study.

Adult↗

PubMed1998

Small GTP-binding proteins in the brain-corpus cardiacum-corpus allatum complex of the silkworm, Bombyx mori: involvement in the secretion of prothoracicotropic hormone.

At least three GTP-binding proteins (G-proteins), 28, 25, and 21 kDa, were found in the brain-corpus cardiacum-corpus allatum complex (BR-CC-CA) of the silkworm, Bombyx mori. They bound to GTP and GDP specifically among nucleotides tested, indicating that these proteins are small G-proteins. The 25 kDa G-protein showed a cross-reactivity to anti-rab3A antibody, while it did not cross-react with anti-rhoA, rab3B, and anti-ras antibodies. On the other hand, the 28 and 21 kDa G-proteins showed no cross-reactivity to any of those antibodies tested. Immunoblot analysis using the anti-rab3A antibody demonstrated that the 25 kDa G-protein was detected preferentially in the BR-CC-CA, and to some extent in the suboesophageal ganglion, but not in the salivary gland, fat body, prothoracic gland, and oesophagus. These results suggested that the 25 kDa G-protein was a member of the rab family of G-proteins. Furthermore, 1 mM GTP gamma S capable of activating G-proteins induced BR-CC-CA to release PTTH under the conditions that stimulation of the PTTH release with hetero-trimeric G-protein was suppressed. These results indicated that the small G-proteins may possibly contribute to PTTH release in Bombyx mori.

Animals↗

PubMed1998

The incidence and characteristics of silent cerebral infarction in elderly diabetic patients: association with serum-soluble adhesion molecules.

The purpose of this study was to investigate the relationship between complications arising from silent cerebral infarction (SCI) and changes in the levels of serum-soluble adhesion molecules in 82 elderly diabetic patients aged 60 years and older. SCI was found in 43 % of the 82 patients, with incidence increasing in relation to age. The prevalence of SCI was higher in subjects with hypertension, poor metabolic control and increased fibrinolysis. The levels of soluble intercellular adhesion molecule-1 (sICAM-1), vascular cell adhesion molecule-1 (sVCAM-1) and E-selectin (sE-selectin) were higher in diabetic patients than in non-diabetic subjects (p < 0.05, p < 0.001, and p < 0.05, respectively). Also, sICAM-1 and sVCAM-1 were found at increased levels in diabetic patients with SCI compared to those without SCI (p < 0.01 and p < 0.05, respectively). In particular, the level of sICAM-1 was increased in patients with SCI due to perforating arterial occlusion, while the level of sVCAM-1 was increased in patients with SCI due to cortical arterial occlusion. However, no significant difference was found in sE-selectin levels. Overall average of the intima and media thickness (IMT) of the common carotid arteries increased with age. IMT proved to be greater in patients with SCI than in patients without SCI (p < 0.05), and showed a weak but significant positive correlation with sVCAM-1, while no correlation was found with either sICAM-1 or sE-selectin levels. In conclusion, measurement of serum adhesion molecules may be useful for diagnosing the early stages of brain damage and for prophylactic treatment which may prevent the onset or progression of SCI.

Aged↗

PubMed1998

Effects of isoenzyme selective phosphodiesterase inhibitors on bacterial lipopolysaccharide-induced bronchial hyperreactivity in guinea pigs.

1. Effects of cilostazol and vesnarinone (selective PDE III inhibitors), rolipram (selective PDE IV inhibitor) and theophylline (nonselective PDE inhibitor) on LPS-induced bronchial hyperreactivity were investigated in guinea pigs. 2. Cilostazol (50 mg/kg PO), vesnarinone (100 mg/kg PO), rolipram (20 mg/kg PO), and theophylline (50 mg/kg PO), significantly inhibited bronchial hyperreactivity to acetylcholine (Ach) and TNF release into bronchoalveolar lavage fluid following LPS exposure. None of these compounds influenced neutrophil influx into bronchoalveolar lavage fluid. 3. Rolipram and theophylline antagonized Ach-induced bronchoconstriction in normal guinea pigs.

Acetylcholine↗

PubMed1998

Lipitoids--novel cationic lipids for cellular delivery of plasmid DNA in vitro.

BACKGROUND: Although synthetic nonviral vectors hold promise for the delivery of plasmid DNA, their gene-transfer efficiencies are far from matching those of viruses. To systematically investigate the structure-activity relationship of cationic lipids, a small library of cationic lipid-peptoid conjugates (lipitoids) was synthesized. The compounds were evaluated for their ability to form complexes with plasmid DNA and to mediate DNA transfer in vitro. RESULTS: Lipid-peptoid conjugates were conveniently prepared in high yield using solid-phase synthesis. Several lipitoids condensed plasmid DNA into 100 nm spherical particles and protected the DNA and DNase digestion. A subset of lipitoids with a repeated (aminoethyl, neutral, neutral) sidechain trimer motif conjugated with dimyristoyl phosphatidyl-ethanolamine (DMPE) mediated DNA transfer with high efficiency. CONCLUSIONS: Automated solid-phase synthesis of cationic lipids allowed the rapid synthesis of a diverse set of transfection reagents. The most active compound DMPE-(Nae-Nmpe-Nmpe)3 (Nae, N-aminoethyl glycine; Nmpe, N-p-methoxyphenethyl-glycine) is more efficient than lipofectin or DMRIE-C (two commercial cationic lipid transfection reagents) and is active in the presence and absence of serum. The activity in the presence of serum suggests potential for applications in vivo.

3T3 Cells↗

PubMed1998

Nitric oxide synthase and NADPH-diaphorase in neurons of the rat, dog and guinea pig nodose ganglia.

Localization of nitric oxide synthase (NOS) in the nodose ganglia of the dog, rat and guinea pig was investigated. A double-staining technique of NOS immunohistochemistry and NADPH-diaphorase (NADPHd) histochemistry was used; then the ratio of NADPHd-positive and NOS-positive cells to the total cells was calculated. The distribution of positive cells within the canine nodose ganglion was also investigated. NADPHd-positive neurons were detected in all the ganglia. Three intensities of reactivity to NADPHd histochemistry (strong, weak or negative) were detected in the neurons of all three species. There were more cells that stained strongly for NADPHd in the rat, but fewer in the dog and guinea pig, indicating that a species difference may exist. NADPHd-positive neurons were less abundant in the rostral third of the canine nodose ganglion than in the middle or caudal thirds. NADPHd reactivity was completely co-localized to the cells that demonstrated neuronal NOS immunoreactivity in the canine nodose ganglion. Thus, NADPHd histochemical reactivity may be a reliable marker of NOS in the nodose ganglion.

Animals↗

PubMed1998

Purification and some characteristics of phosphatase of a psychrophile.

The phosphatase of a psychrophile was purified by ammonium sulfate fractionation, and a sequence of chromatographies on DEAE-Cellulofine, butyl-Cellulofine, Sephacryl S-100, and Mono-Q columns. The purified enzyme preparation was found to be electrophoretically homogeneous on native- and SDS-PAGE, and its molecular mass was determined to be 38.4 kDa by MALDI-TOF mass spectrometry. Maximal activity was observed at 30 degrees C and pH 6.0. Furthermore, the activity of this enzyme at 0 and 5 degrees C was 27 and 28%, respectively, of that at 30 degrees C. The enzyme was stable in the pH range of 6.0 to 8.0 and up to 20 degrees C. The enzyme was affected by metal ions; the activity was enhanced by Mg2+ and Ca2+ ions, but depressed by Zn2+ ions. Analysis of the amino acid composition indicated that this phosphatase contains no S-S bond, and only a few prolyl residues necessary to retain the rigid structure of a protein molecule. The phosphatase shows typical features of a cold enzyme; high catalytic activity at low temperature and rapid inactivation at an intermediate temperature.

Animals↗

PubMed1998

A study of the proliferative activity of the long junctional epithelium using argyrophilic nucleolar organizer region (AgNORs) staining.

The proliferative activity of the long junctional epithelium (LJE) in rats was examined using stains for argyrophilic proteins of the nucleolar organizer region (AgNORs protein). The LJE was experimentally produced by insertion of a rubber piece between maxillary molars for 1 wk. After removal of the rubber, the length and AgNORs parameters of the LJE were measured and analyzed statistically. The LJE widely covered the apical side of the exposed root surface 4 wk after the removal. Its length was longest after 4 and 8 wk; it became shorter subsequently. The AgNORs were visible as black dots of various sizes and numbers on the sections. A high potential for proliferation was obvious in the LJE after 4 wk and was maintained until 12 wk after the removal. The AgNORs ratio on the connective tissue interface of the LJE was about twice of that of normal junctional epithelium after 4-12 wk. The proliferative activity on the root surface side was slightly increased after 4 wk. There was no significant difference in proliferative activity between the coronal and apical sides. These results suggest that the proliferative activity of the LJE is maintained continuously at a high level on the connective tissue interface supplying the epithelial cells. Basal cells proliferate at the connective tissue interface of the LJE, migrate directly to the root surface or via the apical portion and finally desquamate from the surface of the epithelium.

Animals↗

PubMed1998

Calcitonin gene-related peptide-like immunoreactive motoneurons innervating the canine intrinsic laryngeal muscles.

The percentage of calcitonin gene-related peptide (CGRP)-immunoreactive motoneurons in the nucleus ambiguus innervating the intrinsic laryngeal muscles of colchicine-treated dogs was examined by using cholera toxin B subunit-gold (CTBG) as a retrograde tracer and by immunohistochemistry. Neurons that were labeled with CTBG from the cricothyroid muscle were located in the ventromedial division of the rostral part of the nucleus ambiguus, and the ratio of CGRP-positive neurons was 93.0%. Neurons labeled with CTBG from the thyroarytenoid muscle were located in the dorsal division of the medial part of the nucleus ambiguus, and the percentage of neurons with CGRP was 71.4%. Neurons labeled with CTBG from the posterior cricoarytenoid muscle were located in the ventral division of the medial part of the nucleus ambiguus, and the percentage of CGRP-positive neurons was 85.5%. These findings suggest that the innervation and/or the neurotrophic mechanism involving CGRP for each intrinsic laryngeal muscle is different.

Animals↗

PubMed1998

Molecular cloning of cDNA for BRab from the brain of Bombyx mori and biochemical properties of BRab expressed in Escherichia coli.

From a brain cDNA library of Bombyx mori, we cloned cDNA for BRab, which encoded a 202-amino-acid polypeptide sharing 60-80% similarity with rab1 family members. To characterize its biochemical properties, cDNA for BRab was inserted into an expression vector (pGEX2T) and expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein. The recombinant protein was purified to homogeneity with glutathione S-Sepharose. The purified GST-BRab bound [35S]-GTP gamma S and [3H]-GDP with association constants of 1.5 x 10(6) M-1 and 0.58 x 10(6) M-1, respectively. The binding of [35S]-GTP gamma S was inhibited with GTP and GDP, but with no other nucleotides. The GTP-hydrolysis activity was evaluated to be 5 m mole/min/mole of BRab. In the presence of 6 mM MgCl2, bound [35S]-GTP gamma S and [3H]-GDP were exchanged with GTP gamma S most efficiently. These results suggest that BRab, having a higher affinity for GTP than GDP, converts from the GTP-bound state into the GDP-bound state by intrinsic GTP hydrolysis activity and returns to the GTP-bound state with the exchange of GDP with GTP.

Amino Acid Sequence↗

PubMed1998

Advanced nasopharyngeal carcinoma treated with chemotherapy and radiotherapy: distant metastasis and local recurrence.

One hundred and twenty-nine patients with NPC treated at the Department of Radiology, Chiba University Hospital and Keio University Hospital from 1980 through 1993 were selected for this study. Forty-four patients received cisplatin (CDDP)- or carboplatin-based chemotherapy, and 58 patients received adriamycin (ADM)- and/or 5-FU-based chemotherapy. The remaining 27 patients were treated with radiotherapy alone. The median radiation dose to the nasopharyngeal region was 64 Gy, and to the initially involved cervical node, 60 Gy. The 5 year survival rates for the CDDP, the ADM/5-FU and the radiation alone groups were 61%, 47% and 42%, respectively. The cumulative incidences of local control in the CDDP, the ADM/5-FU and the radiation alone groups were 77%, 49% and 53% respectively. The CDDP group achieved the significantly better local control (CDDP vs ADM: p=0.001). The overall incidence of distant metastases was 54% in the CDDP group. On the other hand, it was 24% in the ADM/5-FU group and 22% in the radiation alone group (CDDP vs ADM: p=0.048). While the locoregional control rate was significantly better in the CDDP given group, more distant metastases were seen in this group.

Adolescent↗

PubMed1998

[Easy graphical display of beam directions in three-dimensional converging radiation therapy: proposal for a radiation map].

In stereotactic radiosurgery, non-coplanar isocentric beams are employed to concentrate the dose distribution on the planning target volume (PTV). However, the directions of incident beams must be determined with great care by using a digitally reconstructed beam's eye view (BEV) to prevent the irradiation of organs at risk. We present a new method of 2-dimensional graphical representation (radiation map) to facilitate the understanding of 3-dimensional relationships between incident beams and critical organs. After determining the isocenter and beam diameter, beam directions and critical organs are projected onto the imaginary sphere centered on the isocenter. The coordinate of the beam directions and the organs at risk can be expressed by latitude and longitude on the sphere. The contours of the organs at risk are displayed with a margin of the the radius of the radiation beam. Mirror images of the critical organs are also displayed to prevent irradiation by the opposing beams. The radiation map could be produced within 5 minutes using a workstation. Radiation maps, like DVH, will be very useful in the evaluation of radiation treatment planning.

Radiosurgery↗

PubMed1997

Developmental exposure to diethylstilbestrol elicits demethylation of estrogen-responsive lactoferrin gene in mouse uterus.

Alteration of DNA demethylation in five CpG sites (-547, -533, -475, -464, and -454) immediately upstream from the estrogen response element of lactoferrin promoter was determined in the uteri of immature (17-day-old) and mature (21- and 30-day-old) mice treated neonatally with DES. Only the CpG/-464 was found to be abnormally demethylated by diethylstilbestrol (DES) treatment in the mature uteri. This abnormal demethylation occurred in specific response to DES in neonatal mice, because DES injected into the 30-day-old mature mice did not demethylate CpG/-464. This site, however, remained methylated in the neonatally DES-treated/ovariectomized mice, indicating that this DES-elicited demethylation is under hormonal control. Thus, neonatal DES treatment appeared to imprint an abnormal, site-specific demethylation of CpG/-464, which requires ovarian hormones to occur in adult mice. Moreover, the demethylation was maintained in uterine tumors of the neonatally DES-treated mice. This mode of demethylation is reminiscent of uterine tumor formation, which also depends on both neonatal DES exposure and ovarian hormone stimulation in adulthood. Thus, neonatal DES treatment may induce tumor formation as well as demethylation through a common cellular process.

Age Factors↗