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Biomedical subjects

Y Noda

Publications and source records attributed to Y Noda.

At least 271 records · Page 15Linked to original sources

[Malignant fibrous histiocytoma of the lung].

A 68-year-old man had a malignant fibrous histiocytoma of the lung that was found at autopsy. The patient was admitted to our hospital because of exertional dyspnea. A chest X-ray film and chest CT scan showed atelectasis in the left upper lobe. Examination with a fiberoptic bronchoscope revealed a necrotic mass in the left mainstem bronchus. Microscopic examination of a biopsy specimen disclosed several atypical giant cells but was not diagnostic. He underwent chemotherapy with CBDCA, IFM and VP-16, because a malignant tumor of the lung was strongly suspected. After three cycles of chemotherapy, the tumor had shrunk, as demonstrated on repeated bronchoscopy. The patient's condition improved temporarily, but he began to complain of dyspnea, and died of respiratory insufficiency despite radiotherapy. Postmortem examination was done. The tumor in the lung was mainly composed of spindle-shaped fibroblast-like cells and several pleomorphic giant cells with multiple nuclei, with storiform and fascicular patterns. Immunohistochemically, these tumor cells were shown to contain vimentin, alpha-1-antitrypsin, and LN-5. These findings were compatible with malignant fibrous histiocytoma of the lung.

Aged↗

KIF1B, a novel microtubule plus end-directed monomeric motor protein for transport of mitochondria.

To further elucidate the mechanism of organelle transport, we cloned a novel member of the mouse kinesin superfamily, KIF1B. This N-terminal-type motor protein is expressed ubiquitously in various kinds of tissues. In situ hybridization revealed that KIF1B is expressed abundantly in differentiated nerve cells. Interestingly, K1F1B works as a monomer, having a microtubule plus end-directed motility. Our rotary shadowing electron microscopy revealed mostly single globular structures. Immunocytochemically, KIF1B was colocalized with mitochondria in vivo. Furthermore, a subcellular fractionation study showed that KIF1B was concentrated in the mitochondrial fraction, and purified K1F1B could transport mitochondria along microtubules in vitro. These data strongly suggested that KIF1B works as a monomeric motor for anterograde transport of mitochondria.

Amino Acid Sequence↗

[A new cellulose-barium gel for computed tomography of the esophagus].

A new oral contrast agent for CT of the esophagus was evaluated for its ability to coat the esophageal lumen in 12 patients with esophageal diseases. This agent is made of carboxymethyl cellulose sodium, potato starch and low-density barium, and has remarkably high viscosity of 13600 centipoise. The esophagus was divided into proximal, mid, and distal segments to analyze the effectiveness of lumenal opacification. The average percent opacification of these segments was 78.7, 86.4, and 63.4%, respectively, for all patients. This agent is expected to play a possible role in the CT diagnosis of esophageal cancer by helping to determine the indications for radical surgery and/or radiotherapy.

Barium Sulfate↗

[New development of cellulose-ferric ammonium citrate solution as a contrast-enhancement agent for magnetic resonance imaging of esophageal diseases].

For magnetic resonance imaging (MRI) of esophageal diseases, we have developed a new contrast-enhancement agent, cellulose-ferric ammonium citrate solution. Using this agent, the esophageal lumen is easily identified. This agent is expected to play a role in the MRI diagnosis of esophageal cancer and to help determine the indications for radical surgery and/or intracavitary irradiation.

Aged↗

The methyl viologen-resistance-encoding gene smvA of Salmonella typhimurium.

The complete nucleotide sequence of the gene encoding the Salmonella typhimurium methyl viologen-resistant protein, SmvA, similar to QacA (resistance to quaternary ammonium ion) of Staphylococcus aureus, and the surrounding sequences were determined. This indicated that the gene arrangement of S. typhimurium is different from that of Escherichia coli in this region.

Cloning, Molecular↗

Macro-EMG and motor unit recruitment threshold: differences between the young and the aged.

The relationship between macro-EMG (electromyography) and motor unit recruitment threshold was studied in the first dorsal interosseous (FDI) muscle of normal young and aged subjects. During voluntary isometric contraction, motor unit action potentials (MUAP) were collected by a special quadrifilar electrode and decomposed to each MUAP train (MUAPT) using an EMG signal decomposition technique. Macro-EMG was obtained from the electrode shaft, then triggered and averaged for each MUAPT. A positive linear correlation was observed in both the young and aged subjects. However, the correlation coefficients were significantly lower in the aged individuals than in the young individuals.

Action Potentials↗

A complex of GRB2-dynamin binds to tyrosine-phosphorylated insulin receptor substrate-1 after insulin treatment.

Insulin drives the formation of a complex between tyrosine-phosphorylated IRS-1 and SH2-containing proteins. The SH2-containing protein Grb2 also possesses adjacent SH3 domains, which bind the Ras guanine nucleotide exchange factor Sos. In this report, we examined the involvement of another SH3 binding protein, dynamin, in insulin signal transduction. SH3 domains of Grb2 as GST fusion proteins bound dynamin from lysates of CHO cells expressing wild-type insulin receptor (IR) (CHO-IR cells) in a cell-free system (in vitro). Immunoprecipitation studies using specific antibodies against Grb2 revealed that Grb2 was co-immunoprecipitated with dynamin from unstimulated CHO-IR cells. After insulin treatment of CHO-IR cells, anti-dynamin antibodies co-immunoprecipitated the IR beta-subunit and IRS-1, as tyrosine-phosphorylated proteins and PI 3-kinase activity. However, purified rat brain dynamin did not bind directly to either the IR, IRS-1 or the p85 subunit of PI 3-kinase in vitro. Together, these results suggest that in CHO-IR cells, insulin stimulates the binding of dynamin to tyrosine-phosphorylated IRS-1 via Grb2 and that IRS-1 also associates with PI 3-kinase in response to insulin. This complex formation was reconstituted in vitro using recombinant baculovirus-expressed IRS-1, GST-Grb2 fusion proteins and dynamin peptides containing proline-rich sequences. Furthermore, dynamin GTPase activity was found to be stimulated when an IRS-1-derived phosphopeptide, containing the Grb2 binding site, was added to the dynamin-Grb2 complex in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptor Proteins, Signal Transducing↗

Intratumoral DNA heterogeneity correlated with lymph node involvement and surgical staging in epithelial ovarian cancer by flow cytometry.

BACKGROUND: Flow cytometry (FCM)-measured DNA content may be a predictor in the prognosis of ovarian cancer. Multiple specimens taken from the same ovarian tumor may show a variation in DNA content (i.e., intratumoral DNA heterogeneity). We measured the FCM DNA content of multiple specimens from the same tumor in ovarian cancer, and the relationship among DNA ploidy, intratumoral DNA heterogeneity, retroperitoneal lymph node involvement, and surgical staging was evaluated. METHODS: Forty-one patients with primary epithelial ovarian cancer were included in the study. The FCM-measured DNA content of multiple fresh tumor specimens taken from different parts of the same ovarian tumor from each patient was measured. When aneuploidy was observed in at least one specimen from the same tumor, the tumor was defined as an aneuploid tumor. If there were two or more different aneuploid stem lines with a variation of DNA indices (differences of the DNA indices > 0.15) from the same tumor, the presence of intratumoral DNA heterogeneity was defined. RESULTS: Diploid tumor was found in 8 (19.5%) of the 41 patients, and aneuploid tumor with intratumoral DNA heterogeneity was found in 20 (48.8%). None of the eight patients with diploid tumors demonstrated lymph node involvement. In contrast, lymph node involvement was found in 14 (70.0%) of 20 patients with intratumoral DNA heterogeneity. There was a significant different incidence of lymph node involvement between the groups with and without intratumoral DNA heterogeneity (P < 0.01). The incidence of intratumoral DNA heterogeneity significantly correlated with the International Federation of Gynecology and Obstetrics staging (P < 0.01), while that of aneuploid tumor did not. CONCLUSIONS: Intratumoral DNA heterogeneity reflected a malignant potential for lymph node involvement and its progression in epithelial ovarian cancer.

Aneuploidy↗

Partitioning of vasoactive intestinal polypeptide into lipid bilayers.

Incubation of radiolabeled vasoactive intestinal polypeptide (VIP) with preformed lipid vesicles composed of phosphatidylcholine, phosphatidylglycerol and cholesterol resulted in reversible and saturable association of the peptide with the lipid bilayer. The pH-optimum for the reaction was in the physiological range. The vesicle-associated peptide displayed enhanced stability to proteolytic digestion, it was efficiently released into the supernatant by detergent-solubilization of the vesicles but remained vesicle-associated during treatment with agents that disrupt ionic interactions. Peptide binding by electrically neutral vesicles was lower than that by negative vesicles. Electron spin resonance studies with 5-doxylstearic acid or 16-doxylstearic acid labeled vesicles suggested that VIP decreased the fluidity close to the surface of the bilayer and increased the fluidity in its hydrophobic core. These observations suggest that VIP can bind and penetrate lipid bilayers.

Amino Acid Sequence↗

Specificity of trypsin digestion and conformational flexibility at different sites of unfolded lysozyme.

Fourteen tryptic peptides and nine intermediates were identified as products of trypsin digestion of reduced and S-3-(trimethylated amino) propylated lysozyme. Kinetics of the appearance and disappearance of these products were observed by monitoring the peak areas on the chromatogram. In spite of the complicated reaction pathways, kinetics of the digestion of proteins and several intermediate products show simple decay curves with a single rate constant. In this paper, the trypsin susceptibility of the individual cleavage site is defined as a hydrolytic rate constant of the susceptible peptide bond in the presence of 10 nM trypsin. The cleavage sites of unfolded lysozyme are classified into two groups in terms of the trypsin susceptibility: one has a high susceptibility (10-20 h-1) and the other a low susceptibility (1.0-2.0 h-1). In the unfolded state of lysozyme, in conclusion, the region from residues 15 to 61 has a strong resistance to trypsin digestion; on the other hand, the C-terminal half of the polypeptide chain is flexible enough to fit into the active site of trypsin. In addition, six kinds of pentapeptides were synthesized as analogues of lysozyme fragments including Arg 14, Arg 21, Lys 33, Arg 45, Arg 61, and Arg 73. Kinetics of tryptic digestion of them were observed. Both kcat and KM were determined for these synthetic pentapeptides. The susceptibility of each cleavage site in pentapeptides is determined and compared with that corresponding in proteins. The susceptibility is usually higher when the susceptible peptide bond is included in proteins than in pentapeptides, so long as the conformation of peptide chain is flexible. However, susceptibilities of a few sites in proteins are lower than those in pentapeptides. This means that the peptide chains tend to fold locally to prevent trypsin from binding to the sites. It was found that the sites of Arg 21 and Arg 45 are indeed resistant to trypsin, but the site of Lys 33 is not so much, although the hydrolytic rate at Lys 33 itself is extremely slow.

Amino Acid Sequence↗

Relationship between the day of embryo transfer and the outcome in human in vitro fertilization and embryo transfer.

PURPOSE: To clarify the optimal date of embryo transfer (ET), we retrospectively analyzed the relationship between the day of ET and the outcome in human in vitro fertilization and embryo transfer (IVF-ET). METHOD: Of a total of 307 human IVF-ET cycles performed at Kyoto University Hospital between January 1990 and March 1994, we focused on 207 cases of IVF-ET cycles in which two or three good-quality embryos were transferred. These 207 IVF-ET cycles consisted of 54 Day 2 ET cycles, 79 Day 3 ET cycles, 46 Day 4 ET cycles, and 28 Day 5 ET cycles. We compared the pregnancy and live-birth (plus ongoing pregnancy) rates among these four ET groups. RESULTS: The pregnancy rates of ET on Days 2 to 4 were not significantly different, whereas Day 5 ET produced a significantly lower pregnancy rate (Day 2, 29.6%; Day 3, 32.9%; Day 4, 30.4%; Day 5, 10.7%). Similar results were obtained for the live-birth (plus ongoing pregnancy) rates (Day 2, 20.3%; Day 3, 18.9%; Day 4, 17.9%; Day 5, 7.1%). CONCLUSIONS: These results suggest that the day of ET does not fundamentally affect the pregnancy rate in human IVF-ET provided that transfer is made before Day 5.

Embryo Transfer↗

Developmental blockage of mouse embryos caused by fatty acids.

PURPOSE: It has been shown that lipid peroxides derived from polyunsaturated fatty acids (PUFAs) inhibit the proliferation of various cells. In the meantime, it has been suggested that oxidative stress is closely related to the developmental blockage of mammalian embryos cultured in vitro. In this study, we investigated the effects by various fatty acids on mouse embryo development in vitro, and the reversal of these effects by various antioxidants such as superoxide dismutase, ascorbic acid, alpha-tocopherol, uric acid, and ethylenediaminetetraacetic acid. METHODS: Pronuclear and two-cell stage mouse (ICR) embryos were cultured in Biggers-Whitten-Whittingham medium with 0.3% bovine serum albumin alone or complexed with one of the following fatty acids: palmitic, stearic, oleic, linoleic, linolenic, or arachidonic acid. We also measured the fluorescence emission of embryos in media containing various fatty acids in order to investigate the involvement of H2O2 or lipid peroxidation in embryo development. RESULTS: Palmitic acid and PUFAs including linoleic acid inhibited the embryo development. The inhibitory effect of PUFAs was attenuated by adding antioxidants into the media, while the inhibitory effect of palmitic acid was not. Both pronuclear and two-cell stage embryos with PUFAs showed markedly more intensive emissions than those under other conditions. CONCLUSIONS: These results suggest that lipid radicals can easily be generated in early stage embryos and that blastomeres are among the cells vulnerable to the damage by lipid peroxidation.

Animals↗

Vasoactive intestinal peptide encapsulated in liposomes: effects on systemic arterial blood pressure.

The purpose of this study was to determine whether encapsulation of vasoactive intestinal peptide (VIP) in liposomes enhances its vasoactive effects. Liposomes were formed from a solution of VIP in phospholipids and cholesterol, resulting in incorporation of 0.008 mole peptide/mole phospholipid. Leakage of VIP from the liposomes was undetectable over several days of incubation at 4 degrees C in 0.15 M sodium chloride. Under conditions permitting rapid hydrolysis of VIP by trypsin, there was no breakdown of the encapsulated peptide. Increasing concentrations of the liposome-encapsulated VIP administered intravenously to anesthetized hamsters produced a concentration-dependent decrease in the mean arterial blood pressure. The duration and magnitude of the hypotensive effect of the encapsulated VIP was significantly greater (p < 0.05) compared to equivalent concentrations of the unencapsulated peptide. Infusion of empty liposomes was without significant effect on the mean arterial blood pressure. We conclude that encapsulation of VIP in liposomes potentiates the blood pressure-lowering effect of the peptide.

Animals↗

Effects of visible light and other environmental factors on the production of oxygen radicals by hamster embryos.

Previous studies have demonstrated that developing hamster embryos are very sensitive to visible light. In order to elucidate why visible light exerts a toxic effect on hamster embryos, we examined the effect of visible light on the production of hydrogen peroxide (H(2)O(2)) within individual embryos, using a fluorimetric method. In addition, we examined the H(2)O(2) generating capacity of other factors which are known to be related to the in vitro developmental capacity of hamster embryos. One-cell hamster embryos were cultured with 2',7'-dichlorodihydrofluorescin diacetate, and the fluorescence emissions of the H(2)O(2)-dependent oxidative product in the embryos were measured using an Olympus microscopic photometry system. When embryos were exposed to visible light (14,000 lux) for a specified period (0, 0.5, 1, 2 or 3 min) prior to measurement, the fluorescence emissions from embryos increased with the time of exposure to visible light. An exposure of even 0.5 min resulted in a significant increase in hydrogen peroxide. This increase was more rapid in embryos cultured under 20% O(2) than in those cultured under 5% O(2), and the response was quicker than that observed in mouse embryos. The fluorescence emissions from embryos cultured under 5% O(2) were significantly (P<0.001) lower than those from embryos cultured under 20% O(2) in TLP medium. However, the effects of different oxygen tensions on fluorescence emissions were medium-dependent, and were not significant in embryos cultured in HECM-1 medium. The addition of L-cysteine to or elimination of phenol red from the media decreased the fluorescence emissions from embryos (P<0.001), but glucose and phosphate did not affect them. These results suggest that the toxic effect of visible light on the in vitro development of hamster embryos might be due to increased generation of reactive oxygen species, induced by the visible light. This could be one of the explanations for the strict conditions required for overcoming the in vitro developmental block. It is also suggested that the promotive effects of low oxygen culture and L-cysteine on embryo development seem to be derived from their ability to reduce reactive oxygen species.

Journal Article↗

KIF3A is a new microtubule-based anterograde motor in the nerve axon.

Neurons are highly polarized cells composed of dendrites, cell bodies, and long axons. Because of the lack of protein synthesis machinery in axons, materials required in axons and synapses have to be transported down the axons after synthesis in the cell body. Fast anterograde transport conveys different kinds of membranous organelles such as mitochondria and precursors of synaptic vesicles and axonal membranes, while organelles such as endosomes and autophagic prelysosomal organelles are conveyed retrogradely. Although kinesin and dynein have been identified as good candidates for microtubule-based anterograde and retrograde transporters, respectively, the existence of other motors for performing these complex axonal transports seems quite likely. Here we characterized a new member of the kinesin super-family, KIF3A (50-nm rod with globular head and tail), and found that it is localized in neurons, associated with membrane organelle fractions, and accumulates with anterogradely moving membrane organelles after ligation of peripheral nerves. Furthermore, native KIF3A (a complex of 80/85 KIF3A heavy chain and a 95-kD polypeptide) revealed microtubule gliding activity and baculovirus-expressed KIF3A heavy chain demonstrated microtubule plus end-directed (anterograde) motility in vitro. These findings strongly suggest that KIF3A is a new motor protein for the anterograde fast axonal transport.

Animals↗

A novel microtubule-based motor protein (KIF4) for organelle transports, whose expression is regulated developmentally.

To understand the mechanisms of transport for organelles in the axon, we isolated and sequenced the cDNA encoding KIF4 from murine brain, and characterized the molecule biochemically and immunocytochemically. Complete amino acid sequence analysis of KIF4 and ultrastructural studies of KIF4 molecules expressed in Sf9 cells revealed that the protein contains 1,231 amino acid residues (M(r) 139,550) and that the molecule (116-nm rod with globular heads and tail) consists of three domains: an NH2-terminal globular motor domain, a central alpha-helical stalk domain and a COOH-terminal tail domain. KIF4 protein has the property of nucleotide-dependent binding to microtubules, microtubule-activated ATPase activity, and microtubule plus-end-directed motility. Northern blot analysis and in situ hybridization demonstrated that KIF4 is strongly expressed in juvenile tissues including differentiated young neurons, while its expression is decreased considerably in adult mice except in spleen. Immunocytochemical studies revealed that KIF4 colocalized with membranous organelles both in growth cones of differentiated neurons and in the cytoplasm of cultured fibroblasts. During mitotic phase of cell cycle, KIF4 appears to colocalize with membranous organelles in the mitotic spindle. Hence we conclude that KIF4 is a novel microtubule-associated anterograde motor protein for membranous organelles, the expression of which is regulated developmentally.

Amino Acid Sequence↗

Lethal midline granuloma in Okinawa with special emphasis on polymorphic reticulosis.

Lethal midline granuloma (LMG) is a clinical term used to describe a condition which may be manifested histologically as Wegener's granulomatosis (WG), polymorphic reticulosis (PR), and malignant lymphoma (ML). WG is an inflammatory disease, and PR and ML are considered to represent a neoplastic proliferation of lymphoreticular cells. In this report, twenty-two cases of LMG in Okinawa were examined. The frequency of LMG per 100,000 outpatients of the ear, nose and throat clinic in Okinawa was 67, and the higher frequency of PR (27) and ML (34) in Okinawa than in other districts of Japan was characteristic. Polymerase chain reaction, in situ hybridization, and immunohistochemical studies showed that the proliferating cells in PR were CD43+ and simultaneously contained Epstein-Barr viral genome in their nuclei. The higher frequency of PR and ML in Okinawa is discussed in conjunction with a review of pertinent literature: multiple factors including genetic, viral environmental, and socioeconomic factors seem to affect the frequencies of these diseases.

Adult↗