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Biomedical subjects

N Yoshida

Publications and source records attributed to N Yoshida.

At least 127 records · Page 7Linked to original sources

Comparative study of the molecular mechanisms of oocyte maturation in amphibians.

Maturation-promoting factor (MPF), a complex of Cdc2 and cyclin B, is the final inducer of oocyte maturation. Its activity is controlled by inhibitory phosphorylation of Cdc2 on Tyr15/Thr14 and activating phosphorylation on Thr161. Full-grown immature oocytes of the African clawed frog Xenopus laevis contain inactive MPF (pre-MPF) that comprises cyclin B-bound Cdc2 phosphorylated on Tyr15/Thr14 and Thr161. The synthesis of Mos, but not cyclin B, after stimulation by the maturation-inducing steroid progesterone, is believed to be necessary for initiating Xenopus oocyte maturation through Tyr15/Thr14 dephosphorylation of pre-MPF. In contrast, amphibians other than Xenopus (and also fishes) employ a different mechanism. Full-grown immature oocytes of these species contain monomeric Cdc2 but not cyclin B. MPF is formed after hormonal stimulation by binding of the newly produced cyclin B to the pre-existing Cdc2 and is immediately activated through Thr161 phosphorylation. Mos/MAP kinase is neither necessary nor sufficient for initiating maturation in fishes and amphibians except for Xenopus. We propose a new model of MPF formation and activation during oocyte maturation that is applicable to all amphibians (as well as fishes), based on a novel concept that pre-MPF is an artificial molecule that is not essential for inducing oocyte maturation.

Amphibians↗

Sound conditioning reduces noise-induced permanent threshold shift in mice.

The phenomenon of conditioning-related protection, whereby prior exposure to moderate-level, non-traumatic, sound protects the ear from subsequent traumatic exposure, has been documented in a number of mammalian species. To probe the molecular mechanisms underlying this effect, the mouse would be a useful model; however, a previous study reported no conditioning effects in this species (Fowler et al. , 1995). In our study, mice (CBA/CaJ) were exposed to a traumatic octave-band noise (8-16 kHz at 100 dB SPL for 2 h) with, or without, prior exposure to a sound-conditioning protocol consisting of exposure to the same noise band at lower sound pressure levels. Two conditioning protocols were investigated: one (81 dB SPL for 1 week) was analogous to those used in other conditioning studies in mammals; the second was significantly shorter (89 dB SPL for 15 min). Noise-induced permanent threshold shift (PTS) was assessed in a terminal experiment, after the traumatic exposure, via compound action potentials. Neither conditioning protocol elevated threshold, indeed both protocols increased amplitudes of distortion product otoacoustic emissions when animals were conditioned but not traumatized. Both conditioning exposures significantly reduced PTS from the subsequent traumatic exposure, compared to groups exposed without prior conditioning. Protective effects of 15-min conditioning were maximal when the condition-trauma interval was 24 h; protection disappeared when the traumatic exposure was presented 48 h after conditioning. These data are consistent with the view that protein synthesis is required for expression of the protective effect. The enhancement of distortion products in the condition-only state suggests that conditioning changes outer hair cell function.

Animals↗

Acoustic injury in mice: 129/SvEv is exceptionally resistant to noise-induced hearing loss.

129/SvEv is an inbred mouse strain popular for use in genetic knockout studies. Here, we compare normal auditory function and vulnerability to acoustic injury in wild-type mice of the 129/SvEv vs. CBA/CaJ strains. Compound action potentials (CAPs) and distortion product otoacoustic emissions (DPOAEs) showed slightly higher thresholds for 129/SvEv re CBA/CaJ, especially at frequencies >20 kHz. Middle-ear motion (i.e. umbo velocity) was similar in the two strains; although frequencies >20 kHz could not be evaluated. Permanent threshold shift (PTS) and hair cell losses, measured 1 week after high-intensity exposure to an 8-16 kHz noise band, were smaller in129/SvEv at all exposure levels and durations from 97 dB SPLx2 h to 106 dB SPLx8 h. Furthermore, PTS growth with increasing exposure energy was slower in 129/SvEv (<2 dB/dB) than CBA/CaJ (9 dB/dB). These data suggest that the vulnerability differences lie in the inner ear, not the middle ear. Several 129/Sv substrains show age-related hearing loss (AHL): 129/SvEv has not yet been evaluated (Zheng, Q.Y., Johnson, K. R., Erway, L.C., 1999. Assessment of hearing in 80 inbred strains of mice by ABR threshold analyses. Hear. Res. 130, 94-107). Thus, although other strains with AHL, e.g. C57Bl/6J, show increased vulnerability to noise-induced hearing loss (NIHL), pairing of AHL and NIHL vulnerabilities may not be obligatory.

Aging↗

Lipoprotein(a) is a risk factor for occurrence of acute myocardial infarction in patients with coronary vasospasm.

OBJECTIVES: The purpose of this study is to determine whether lipoprotein(a) (Lp[a]) is an independent risk factor for coronary spasm and occurrence of acute myocardial infarction (AMI) in patients with coronary spasm. BACKGROUND: Although elevated serum Lp(a) levels are known to be associated with coronary atherosclerosis and AMI, the association between the elevated level of this lipoprotein and coronary spasm remains to be elucidated. METHODS: Serum Lp(a) levels were measured using a latex immunoassay in 77 patients with coronary spasm but without a significant (>75%) fixed coronary stenosis, including 16 with prior myocardial infarction (MI), in 177 patients with a fixed stenosis but without rest angina, including 114 with prior MI and in 81 control subjects without coronary artery disease. RESULTS: The serum Lp(a) level in patients with coronary spasm (median; 17 mg/dl) was higher (p < 0.01) than in control subjects (12 mg/dl) but lower (p < 0.01) than in patients with a fixed stenosis (23 mg/dl). The incidence of subjects with higher (>25 mg/dl) serum Lp(a) levels was higher in patients with a fixed stenosis (46%, p < 0.01) but not in patients with coronary spasm (27%), compared with control subjects (21%). Among the patients with coronary spasm, the incidence of higher Lp(a) levels was higher in patients with than in those without a history of prior MI (56% vs. 21%, p < 0.05). The patients with higher Lp(a) levels had a higher incidence of prior MI than those without (41% vs. 13%, p < 0.05). The multivariate analysis confirmed that higher serum Lp(a) level is an independent determinant for prior MI in these patients (odds ratio, 4.19; 95%, confidence interval, 1.03 to 17.00). CONCLUSIONS: Elevated serum level of Lp(a) was found to be associated with a history of prior MI in patients with coronary spasm, suggesting that Lp(a) may play an important role in the genesis of thrombotic coronary occlusion and the occurrence of AMI subsequent to coronary spasm.

Adolescent↗

Ets family transcription factor ESE-1 is expressed in corneal epithelial cells and is involved in their differentiation.

Involvement of an epithelium-specific transcription factor ESE-1/ESX/ELF3/jen (ESE-1) in corneal epithelial cell differentiation was investigated. ESE-1 was reported to be induced during terminal differentiation of the epidermis and primary keratinocytes and to transactivate target genes through ets binding sites. However, its expression and function in corneal epithelium have not been examined. We report here that ESE-1 is upregulated upon differentiation in mouse corneal epithelium and in immortalized human corneal epithelial cells (HCE). ESE-1 transactivates through the regulatory element of cornea-specific K12 keratin. Moreover, introduction of ESE-1 antisense RNA in HCE cells affect their differentiation. These data suggest the involvement of ESE-1 in differentiation of corneal epithelial cells.

Animals↗

Acute Trypanosoma cruzi infection is associated with anemia, thrombocytopenia, leukopenia, and bone marrow hypoplasia: reversal by nifurtimox treatment.

In the present work we show that acute infection of C3H mice with the CL strain of Trypanosoma cruzi is characterized by an exponential growth of parasites and high mortality accompanied by anemia, thrombocytopenia, leukopenia, and bone marrow hypoplasia. Administration of nifurtimox, a trypanocydal drug currently in clinical use at different days postinfection, modulates parasitemia and prevents mortality. More importantly, none of blood and bone marrow alterations were observed in nifurtimox-treated animals when treatment was initiated early in infection, one or seven days postinoculation. The bone marrow alterations were characterized by a decrease in the total number cells as well in the number of megakaryoblasts and erythroblasts. Transfer experiments of bone marrow cells from infected mice to noninfected lethally irradiated recipients revealed a poor marrow-repopulating activity. The colony forming units-spleen assay confirmed the depression of committed clonal progenitors cells and revealed a decreased number of granulocyte/macrophage, megacariocyte and erythrocyte colonies. In summary, this is the first report showing that acute T. cruzi infection results in profound alterations of the hematopoietic system and that these alterations can be prevented by nifurtimox treatment.

Anemia↗

A new method for qualitative and quantitative evaluation of tooth displacement under the application of orthodontic forces using magnetic sensors.

The purpose of this study was to develop a new method for the quantitative and qualitative measurement of tooth displacement under load and to clarify the relationship between the force system acting on a tooth and a resultant movement. A combination of eight magnetic sensors and a magnet was employed to measure three-dimensional displacement. Two sets of sensors and a magnet were placed in front and at the back of a front tooth subjected to orthodontic forces. Magnets and sensors were fixed to the subject tooth and posterior teeth, respectively. Sensors detected displacements of two magnets extended from the subject tooth separately in real time. The tooth trajectories projected on the sagittal plane were studied. From displacements of two magnets, angle of tooth rotation, the position of center of rotation and the amount and direction of displacement of an arbitrary point on the tooth could be calculated with high accuracy. Also, the location of center of resistance could be determined. It was found that the center of rotation is moving substantially even with a small difference in the level of force application. Furthermore, a small amount of displacement of the center of resistance was observed with varying the force magnitude.

Biomechanical Phenomena↗

Polyol production by culture of methanol-utilizing yeast.

Four methanol-utilizing yeasts, Candida boidinii, Hansenula polymorpha, Hansenula ofunaensis, and Pichia pinus, produced polyols from corresponding sugars in a methanol medium. H. polymorpha produced larger amounts of xylitol than the other yeasts. Productivity was the highest at pH 8 when 5 g (dry)/l cultured cells were incubated with 2.5 g/l urea as the nitrogen source in a medium containing 1% (v/v) methanol and 1 g/l MgSO4.7H2O. Under these conditions, 57 g/l xylitol was obtained from 110 g/l D-xylose after 3 d of cultivation. The largest amount of xylitol (58 g/l; yield, 0.62 g/g) was produced from 125 g/l, D-xylose and 5% (w/v) glycerol instead of methanol after 4 d of cultivation.

Journal Article↗

Origin of fan palm (Livistona chinensis R. Br. var. subglobosa Becc.) in Aoshima, Japan.

RAPD and RFLP analyses were carried out to determine the origin of Livistona chinensis R. Br. var. subglobosa Becc. from Iriomotejima, Ishigakijima, Okinawa, Yakushima, Tanegashima, Cape Sata, Cape Toi, Tsukishima, and Aoshima, Japan. RAPD data obtained using 5 random primers were clustered using UPGMA or the neighbor-joining method. Each population was classified into three clusters based on the phylogenetic tree. L. chinensis plants from Yakushima, Tanegashima and Cape Sata belonged to the isologous cluster, and those from Ishigakijima and Okinawa are contained in a different cluster. L. chinensis from Okinawa and Ishigakijima could be differentiated by being the oldest based on the genetic distance. The area that covers Ishigakijima and Okinawa is thought to be the origin of L. chinensis. L. chinensis plants from Iriomotejima were contained in the same cluster as those from Aoshima. The phylogenetic trees constructed by both RAPD and RFLP analyses indicate the possibility that seeds or green woods of L. chinensis were dispersed by tidal current from the south field around Irlomotejima, and they were washed to Aoshima and established gradually. Therefore, we support the drifting-ashore-naturalized-plant hypothesis on the origin of L. chinensis in Aoshima.

Journal Article↗

Effect of vitamin E on aspirin-induced gastric mucosal injury in rats.

We investigated the effect of vitamin E on aspirin-induced gastric mucosal injury in rats. Twenty-eight male Sprague-Dawley rats were divided into four groups and were fed for 20 weeks with a diet containing <0.1 mg/100 g of alpha-tocopherol (vitamin E-deficient), 2 mg/100 g of alpha-tocopherol (normal and vitamin E-sufficient), or 50 mg/100 g of alpha-tocopherol (vitamin E-supplemented). In vitamin E-deficient rats, oral administration of aspirin (200 mg/kg) plus HCI created more severe hemorrhagic erosions than in other rats. Vitamin E-deficient rats had higher levels of thiobarbituric acid reactive substances, myeloperoxidase activity, and cytokine-induced neutrophil chemoattractant in the gastric mucosa. Flow cytometry showed that CD18 expression on stimulated neutrophils was higher in vitamin E-deficient rats than in vitamin E-supplemented rats. These results suggest that vitamin E protects against aspirin-induced gastric mucosal injury by inhibiting lipid peroxidation and accumulation of activated neutrophils.

Animals↗

Role of loop D of the alpha7 nicotinic acetylcholine receptor in its interaction with the insecticide imidacloprid and related neonicotinoids.

1. The nitroguanidine insecticide imidacloprid along with a second generation of related compounds including nitenpyram, all nicotinic acetylcholine (ACh) receptor ligands, are used increasingly in many countries. Site-directed mutagenesis and heterologous expression in Xenopus laevis oocytes have been deployed to investigate mutants (G189D and G189E) of the chicken alpha7 homomer-forming nicotinic receptor subunit which are predicted to enhance the negative charge at the negative subsite (loop D) of the ACh binding site. 2. Xenopus oocytes expressing wild-type alpha7 nicotinic receptors respond to imidacloprid with rapid inward currents. Imidacloprid and nitenpyram are partial agonists, whereas ACh, (-)-nicotine and (+)-epibatidine are full agonists. 3. Compared to wild-type alpha7, the mutant G189D and G189E receptors are much less sensitive to the insecticides, whereas their sensitivity to (-)-nicotine, ACh and (+)-epibatidine is only slightly reduced. In contrast, G189N and G189Q mutants are sensitive not only to ACh, (-)-nicotine and (+)-epibatidine, but also to the two insecticides. Thus reduction of the insecticide-sensitivity by the mutations G189D and G189E are attributed to an increase in negativity of loop D. Desnitro-imidacloprid (DN-IMI), an imidacloprid derivative lacking the nitro group is a potent agonist on the G189D and G189E mutants suggesting an important role of loop D in nicotinic receptor interactions with the nitro group of nitroguanidine insecticides.

Animals↗

A mediator-type biosensor as a new approach to biochemical oxygen demand estimation.

A novel biosensor for the determination of biochemical oxygen demand (BOD) was developed using potassium hexacyanoferrate(III) [HCF(III)] as a mediator. The sensor element consists of a three-electrode system, with both working and counter electrodes compactly integrated as a disposable using etching and electroplating processes. Pseudomonas fluorescens biovar V (isolated from a wastewater treatment plant) was immobilized on the surface of the working electrode using poly(vinyl alcohol)-quaternized stilbazol (PVA-SbQ) photopolymer gel. Synthetic wastewater described by the Organization for Economic Cooperation and Development (OECD) was used as a standard solution instead of glucose-glutamic acid synthetic wastewater. The conditions of amperometric measurement were optimized at +600 mV (vs. Ag/AgCl) operating potential, namely 40 mM HCF(III) in a 0.1 M phosphate buffer (pH 7.0) at 20 degrees C. The sensor response was linear from 15 up to 200 mg O l-1 BOD. The response time was 15 min at 200 mg O l-1 BOD. To demonstrate the wide metabolic range of activity of the sensor, the sensor response to 14 substances in four categories of organic compounds was investigated. Further, it was shown that the response of this BOD sensor was not influenced in samples with low concentrations of dissolved oxygen under the measuring conditions used. For real wastewaters, the BOD values were determined using the sensor and compared favorably with those determined by the conventional BOD5 method.

Journal Article↗

A new mechanism for anti-inflammatory actions of proton pump inhibitors--inhibitory effects on neutrophil-endothelial cell interactions.

BACKGROUND: Neutrophil-endothelial cell interactions mediated by adhesion molecules may be involved in gastric mucosal inflammation associated with Helicobacter pylori or nonsteroidal anti-inflammatory drugs. AIM: To investigate the effects of proton pump inhibitors and histamine-2 receptor antagonists (HRA) on neutrophil-endothelial cell adhesive interactions induced by H. pylori water extract (HPE) or interleukin-1beta (IL-1beta). METHODS: Human peripheral neutrophils and umbilical vein endothelial cells were incubated with either proton pump inhibitors (lansoprazole and omeprazole) or HRA (famotidine and ranitidine). Neutrophil surface expression of CD11b and CD18 and endothelial cell intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1) were assessed by flow cytometry and an enzyme immunoassay, respectively. Neutrophil adherence was defined as the ratio of exogenous neutrophils that adhered to the endothelial monolayers. RESULTS: The expression of CD11b and CD18 on neutrophils and neutrophil-dependent adhesion to endothelial cells elicited by HPE were inhibited by lansoprazole and omeprazole at clinical relevant doses, and the expression of ICAM-1 and VCAM-1 on endothelial cells and endothelial-dependent neutrophil adherence induced by IL-1beta were also inhibited by lansoprazole and omeprazole at similar doses. Famotidine and ranitidine had no effect on neutrophil-endothelial cell interactions. CONCLUSIONS: These results indicate that proton pump inhibitors can attenuate neutrophil adherence to endothelial cells via inhibiting the expression of adhesion molecules, suggesting that proton pump inhibitors may have anti-inflammatory activity.

2-Pyridinylmethylsulfinylbenzimidazoles↗

Protective role of intracellular glutathione against nitric oxide-induced necrosis in rat gastric mucosal cells.

BACKGROUND: Nitric oxide synthase activity is increased in the stomach in association with Helicobacter pylori infection and portal hypertension, but the mechanism by which nitric oxide contributes to mucosal damage remains unclear. AIM: To examine whether nitric oxide injures gastric mucosal cells and whether cellular glutathione affects nitric oxide-induced cytotoxicity. METHODS: A confluent monolayer of RGM-1 gastric mucosal cells was exposed to nitric oxide donors (NOC5 or NOC12). Cell viability was determined by trypan blue dye exclusion, lactate dehydrogenase release and supravital staining with Hoechst 33342 and propidium iodide. The kinetics of the reduced/oxidized forms of glutathione were also measured, as well as the effect of glutathione-depletion or glutathione-precursor treatment on nitric oxide-induced cytotoxicity. RESULTS: Excess exogenous nitric oxide produced by NOC5 or NOC12 induced necrosis in RGM-1 cells in a time- and concentration-dependent manner. The level of reduced glutathione drastically decreased prior to the loss of cell viability and remained low, but oxidized glutathione was not affected. Glutathione depletion increased necrosis of both NOCs in an NOC-concentration-related fashion, while pre-treatment with gamma-glutamylcysteine ethyl ester reduced their necrotic susceptibility. CONCLUSION: Exogenous nitric oxide induced necrosis in gastric mucosal cells, and intracellular reduced glutathione protects gastric mucosal cells from damage by nitric oxide.

Animals↗

Forced expression of terminal deoxynucleotidyl transferase in fetal thymus resulted in a decrease in gammadelta T cells and random dissemination of Vgamma3Vdelta1 T cells in skin of newborn but not adult mice.

The repertoire of lymphocyte receptor genes encoded in a germline is further diversified by a number of processes, including the template-independent addition of nucleotides (N regions) by means of terminal deoxynucleotidyl transferase (TdT). Normally, mouse gammadelta T cells in the early fetal thymus, whose T-cell receptor (TCR) genes lack N regions and are encoded by Vgamma3-Jgamma1 and Vdelta1-Ddelta2-Jdelta2 with canonical junctions (invariant Vgamma3Vdelta1), are thought to be the precursors of dendritic epidermal T cells (DETC). We generated mutant mice whose endogenous TdT promoter was replaced with the lck promoter through homologous recombination. These mutant mice expressed TdT in fetal thymus, had abundant N regions and infrequent canonical junctions in gamma and delta rearrangements, and showed a decreased number of gammadelta T cells. Various Vgamma3Vdelta1 T cells, most of which had N regions in their TCR genes, were found to disseminate in the skin of newborn mutant mice, whereas normal numbers of DETCs with the invariant Vgamma3Vdelta1 rearrangement were observed in adult mutants. These data demonstrate that the regulation of TdT expression during fetal development is important for the generation of gammadelta T cells, and that Vgamma3Vdelta1 T cells, which have various junctional sequences in their TCR genes, randomly disseminate in skin, but invariant Vgamma3Vdelta1 T cells have a great advantage for proliferation in skin.

Animals↗

Image processing experiments for computer-based three-dimensional reconstruction of neurones from electron micrographs from serial ultrathin sections.

This study examined an image processing technique that uses a computer to reconstruct a three-dimensional image of neurones from electron micrographs of serial ultrathin sections. The major problems involved were: (a) a distortion of features in electron micrographs; (b) a significant change of cross-section features of neurones in electron micrographs of neighbouring sections; and (c) disagreement between the electron microscopic section face and the coordinate plane desired for the reconstruction. Electron micrographs of a retinal bipolar cell stained with a biotinylated tracer were used. We corrected the distortion of features by means of a warp, a widely used algorithm in morphing image processing. The change of features between neighbouring electron micrographs was minimized by filling the gaps with an interpolated image produced by a dissolve, another algorithm in morphing, as well as the warp. The distortion of the three-dimensional reconstructed image made by piling up features was corrected by making the image with a wire frame model. Furthermore, in order to estimate a closed contour of features, an active contour model, Snakes, was applied to the electron microscope features. Snakes successfully detected the contour of the target feature, but in some electron microscope images broke into the target feature.

Algorithms↗

Non-dependence of cyclin E/Cdk2 kinase activity on the initiation of oocyte maturation in goldfish.

Cdk2 kinase activity increases during oocyte maturation but neither cyclin A nor B is associated with Cdk2 in mature oocytes in goldfish. As a potential Cdk2 partner in meiosis, a cyclin E homolog was isolated from a goldfish oocyte cDNA library. A monoclonal antibody was raised against bacterially produced full-length goldfish cyclin E. Both cyclin E and Cdk2 were already present in immature oocytes and their protein levels did not change remarkably during oocyte maturation. Cyclin E formed a complex mainly with Cdk2 just at the time of germinal vesicle breakdown (GVBD) in association with the increase in Cdk2 kinase activity, although a fraction of cyclin E bound to Cdk(s) other than Cdk2 and Cdc2. Ectopic activation of cyclin E/Cdk2 by the injection of cyclin E messenger RNA (mRNA) into immature oocytes did not induce maturation-promoting factor (MPF) activation and GVBD. Furthermore, inhibition of cyclin E/Cdk2 kinase activity by the injection of p21SDI1 into the oocytes treated with 17alpha,20beta-dihydroxy-4-pregnen-3-one had no effect on MPF activation and GVBD. These results indicate that cyclin E/Cdk2 kinase activity is insufficient and unnecessary for initiating goldfish oocyte maturation.

Amino Acid Sequence↗

Vitamin E and leukocyte-endothelial cell interactions.

Leukocyte-endothelial cell interactions are mediated by adhesion molecules, expression of which is modulated by cytokines and chemical mediators in the early phase of inflammatory and immunologic reactions, including the development of atherosclerosis. Vitamin E is a lipid-soluble antioxidant that is present in all cell membranes at a low concentration and is reported to be an anti-atherogenic agent. Recently, it was reported that vitamin E inhibits the activation of protein kinase C and nuclear factor-kappa B (NF-kappa B). We demonstrated that vitamin E can prevent leukocyte-endothelial cell adhesion by inhibiting signal transduction involved in the surface expression of adhesion molecules by leukocytes and endothelial cells. These results suggest that vitamin E may have a protective effect against the progression of inflammation and atherosclerosis.

Animals↗