Biomedical subjects

S Fujii

Publications and source records attributed to S Fujii.

At least 109 records · Page 6Linked to original sources

PubMed2000

Molecular and crystal structure of galactinol dihydrate [1-O-(alpha-D-galactopyranosyl)-myo-inositol dihydrate].

The crystal structure of galactinol dihydrate has been determined by X-ray diffraction. The crystal belongs to the orthorhombic system, space group P2(1)2(1)2, a = 15.898(6), b = 19.357(5), c = 5.104(4) A, and Z = 4. The structure was refined to R = 0.044 for 1818 observed structure amplitudes. The primary hydroxyl group exhibits twofold orientational disorder. The linkage conformation is close to those of alpha-(1 --> 4) linkages in methyl alpha-maltotrioside tetrahydrate and erlose trihydrate. Although there is no interring hydrogen bond in galactinol, an indirect interring hydrogen bond including a water molecule is present. The observed conformation is additionally stabilized by the indirect interring hydrogen bond. The global minimum in the relaxed-residue energy map based on the MM3(92) force-field is close to the observed conformation in the crystal structure. All hydroxyl, ring and water oxygen atoms are involved in a complex three-dimensional hydrogen-bonding network.

Carbohydrate Conformation↗

PubMed2000

Induction of tryptophan 2,3-dioxygenase in the mouse endometrium during implantation.

Indoleamine 2,3-dioxygenase (IDO) is expressed in trophoblasts and defends the conceptus against rejection by reducing the tryptophan level and suppressing the T cell activity. We isolated a cDNA for tryptophan 2,3-dioxygenase (TDO), another key catabolizing enzyme of tryptophan, from a mouse uterus cDNA library enriched with pregnancy-induced genes. Northern blot and in situ hybridization analyses demonstrated that the TDO mRNA was induced in the decidualized stromal cells around the implanted embryo at the time of implantation. The expression was then upregulated and primarily localized at the mesometrial decidua. TDO mRNA was induced by deciduoma formation as well as embryo transfer but not by ovarian steroid hormones. These findings demonstrated that TDO is induced in the endometrial stromal cells concomitant with decidualization and suggested its involvement in the implantation process by regulating the tryptophan level at the implantation site.

Animals↗

PubMed2000

Antinociceptive effect of cilnidipine, a novel N-type calcium channel antagonist.

We investigated the antinociceptic effects of cilnidipine, a dihydropyridine derivative which acts on both L- and N-type voltage-dependent calcium channels, in mice. Intrathecally injected cilnidipine showed significant analgesic effect in formalin test. Cilnidipine significantly suppressed N-type currents in dorsal root ganglion (DRG) cells. Our findings apparently support the idea that cilnidipine attenuates synaptic neurotransmission by inhibiting N-type calcium channels in DRG neurons.

Animals↗

PubMed2000

Inactivation of alpha7 ACh receptors and activation of non-alpha7 ACh receptors both contribute to long term potentiation induction in the hippocampal CA1 region.

Acute and chronic nicotine exposure differentially facilitate the induction of long-term potentiation (LTP), a synaptic model of learning and memory, in the hippocampal CA1 region. The mechanisms underlying these effects of nicotine, however, are unknown. In the present study, both nicotinic acetylcholine receptor (nAChR) agonists and an alpha7 nAChR antagonist facilitated the induction LTP in the hippocampal CA1 region of naive rat. Furthermore, chronic nicotine treatment lowered the threshold for induction of LTP, and acute application of nicotinic agonists, but not an alpha7 antagonist, further facilitated LTP induction in the chronic-nicotine-treated hippocampus. These results suggest not only that both activation of non-alpha7 nAChRs and inactivation of alpha7 nAChRs contribute to LTP induction, but also that chronic-nicotine-mediated facilitation of LTP induction is due to chronic-nicotine-induced desensitization of alpha7 nAChRs.

Animals↗

PubMed2000

Nicotine reverses GABAergic inhibition of long-term potentiation induction in the hippocampal CA1 region.

Nicotine is known to enhance cognitive function but the mechanism is unknown. The present study examined the modulatory effect of nicotine on the induction of long-term potentiation (LTP), a synaptic model of learning and memory. A weak tetanic stimulation consisting of 20 pulses at 100 Hz induced stable LTP in the hippocampal CA1. The induction of LTP was completely blocked if the tetanus was delivered in the presence of muscimol (2.5 microM), a gamma-aminobutyric acid (GABA) receptor agonist. This inhibition was sensitive to, and reversed by, not only nicotinic acetylcholine receptor (nAChR) agonists (nicotine and epibatidine), but also the alpha7 nAChR-selective antagonist methyllycaconitine (MLA). Furthermore, ACh-puff activation of alpha7 nAChRs on feedforward interneurons induced inhibitory postsynaptic currents in pyramidal cells that were blocked by nicotine or MLA. In addition, nicotine reduced field monosynaptic inhibitory postsynaptic potentials in the presence of MLA. These results suggest not only two pathways of nicotine-induced disinhibition of pyramidal cells, one involving desensitization of alpha7 nAChRs and the other involving non-alpha7 nAChRs, but also two potential mechanisms underlying the modulatory effect of nicotine on LTP induction, both reducing GABAergic inhibition, thereby indirectly increasing the excitability of pyramidal cells.

Aconitine↗

PubMed2000

Cloning and characterization of the gene encoding RNA polymerase sigma factor sigma(54) of deep-sea piezophilic Shewanella violacea.

We have recently reported that a sigma(54)-like factor recognizes a DNA element, designated as region A, upstream of a pressure-regulated operon in piezophilic Shewanella violacea strain DSS12 (Nakasone et al., FEMS Microbiology Lett. 176 (1999) 351-356). In this study, we isolated and characterized the rpoN gene of this piezophilic bacterium. The rpoN gene was found to encode a putative protein consisting of 492 amino acid residues with a predicted molecular mass of 55359 Da. Significant homology was evident comparing the rpoN sequence of S. violacea with that of Escherichia coli (62.8% identity), Vibrio anguillarum (61.7% identity) and Pseudomonas putida (57.0% identity). The DNA-binding domain at the C-terminus of sigma(54) is well conserved in the case of the S. violacea rpoN gene product and the helix-turn-helix motif and the RpoN box are also present. In addition, the conserved glutamine-rich domain is present at the N-terminus. sigma(54) in S. violacea was expressed at a relatively constant level under various growth conditions as determined by both primer extension and Western blotting analyses. By means of a recombinant plasmid, a hexahistidine-tagged derivative of the sigma(54) from strain DSS12 was overexpressed in Escherichia coli and purified to near homogeneity. An electrophoretic mobility shift assay demonstrated that the purified sigma(54) protein specifically recognizes region A in the above-mentioned pressure-regulated operon.

Amino Acid Sequence↗

PubMed2000

Identification of two polyketide synthase gene clusters on the linear plasmid pSLA2-L in Streptomyces rochei.

The 200kb linear plasmid pSLA2-L was suggested to be involved in the production of two macrolide antibiotics, lankamycin (Lm) and lankacidin (Lc), in Streptomyces rochei 7434AN4. Hybridization experiments with the polyketide synthase (PKS) genes for erythromycin and actinorhodin identified two eryAI-homologous regions and an actI-homologous region on pSLA2-L. The nucleotide sequence of a 3.6kb SacI fragment carrying one of the eryAI-homologs revealed that it codes for part of a large protein with four domains for ketoreductase, acyl carrier protein, ketosynthase, and acyltransferase. Gene disruption confirmed that the two eryAI-homologs are parts of a large type-I PKS gene cluster for Lm. A 4.8kb DNA carrying the actI-homologous region contains four open reading frames (ORF1-ORF4) as well as an additional ORF, i.e. ORF5, which might code for a thioesterase. Deletion of the ORF2-ORF4 region showed that it is not involved in the synthesis of Lm or Lc. Thus, it was confirmed that pSLA2-L contains two PKS gene clusters for Lm and an unknown type-II polyketide.

Amino Acid Sequence↗

PubMed2000

Endogenous adenosine regulates the effects of low-frequency stimulation on the induction of long-term potentiation in CA1 neurons of guinea pig hippocampal slices.

A train of low-frequency afferent stimuli (LFS, 1 Hz, 1000 pulses), given 60 min prior to a tetanus (100 Hz, 100 pulses), suppresses the induction of long-term potentiation (LTP) in which a short-term potentiation decreases gradually back to the pre-tetanic level within 40-50 min (LTP suppression). We investigated the effects of adenosine A1 or A2 receptor antagonists (8-cyclopentyltheophylline (8-CPT) and CP-66713, respectively) on LTP suppression in CA1 neurons of guinea pig hippocampal slices. When the LFS was delivered in the presence of 8-CPT (1 microM), LTP suppression was not significantly affected. However, when LFS was delivered in the presence of CP-66713 (10 microM), LTP suppression was inhibited, leading to successful LTP induction. These results indicate that endogenous adenosine, acting via A2 receptors, is involved in the mechanism of LTP suppression.

Adenosine↗

PubMed2000

Up-regulation of crystallin mRNAs in form-deprived chick eyes.

Form-deprivation of chicks during early postnatal development results in ocular enlargement and great myopic refractive error (form-deprivation myopia). Previous studies have indicated that the retina, RPE and choroid play important roles in ocular enlargement in form-deprivation myopia. We aimed to isolate genes up-regulated in the retina-RPE-choroid of form-deprived chick eyes. A suppression subtractive hybridization method was used to compare mRNA expression in the retina-RPE-choroid of form-deprived and control eyes. One up-regulated cDNA was isolated and identified as part of chick delta1-crystallin cDNA. Northern blot and RT-PCR analyses demonstrated that delta1-crystallin mRNA was up-regulated in the retina-RPE at day 7 after form-deprivation treatment. Semi-quantitative RT-PCE analysis of the expression of several transcription factors indicated that Sox1 and Sox3 were upregulated in parallel with delta1-crystallin mRNA in form-deprived eyes. Northern blot analysis demonstrated that alphaA-, betaA3/A1-, betaB1-, and betaB2-crystallin mRNAs were also up-regulated in form-deprived eyes. Although the detailed mechanisms and functions of the crystallin family genes in the retina-RPE-choroid of form-deprived eyes remain unclear, results of our study suggests that form-deprivation affects the expression of these genes in chick eyes.

Animals↗

PubMed2000

Expression of Sonic hedgehog and retinal opsin genes in experimentally-induced myopic chick eyes.

The purpose of this study was to evaluate changes in the expression of different genes in chick retinal tissues after induction of experimental myopia and to evaluate the roles of these genes in the regulation of postnatal eye growth and myopia. Form-deprivation using occlusive goggles and hyperopic defocus by negative spectacle lenses were used to induce myopia in hatched chicks. Expression levels of Sonic hedgehog, its receptor complex, and other retinal cell genes were evaluated by semi-quantitative reverse transcription-polymerase chain reaction. Levels of Sonic hedgehog protein were further evaluated by Western blot analysis. The induction of myopia caused significant increase in expression of Sonic hedgehog mRNA and protein and increased expression of blue and red opsin mRNA. In contrast, the expression of mRNA for Sonic hedgehog receptor complex (Patched-Smoothened), rhodopsin, vimentin, green opsin, violet opsin, and HPC-1 were unaffected by the induction of myopia. The increase in expression of Sonic hedgehog in chick retinas in experimentally-induced myopia suggests involvement in the retina control of postnatal eye growth. Furthermore, Sonic hedgehog may influence the expression of blue and red opsins under myopic conditions.

Animals↗

PubMed2000

Endometrial carcinoma remaining after term pregnancy following conservative treatment with medroxyprogesterone acetate.

BACKGROUND: Successful pregnancies after conservative progestin treatment to young women with endometrial carcinoma have recently been reported. However, it is not known for certain whether the lesion is completely eradicated in such patients. We present a case of residual endometrial carcinoma after term pregnancy which had been treated conservatively before the pregnancy began. CASE: A 28-year-old woman with endometrial carcinoma received conservative treatment with high-dose medroxyprogesterone acetate (MPA) and then conceived. After delivery at term, atypical cells were found in the endometrial curettage specimen. A hysterectomy was performed 6 months after delivery and revealed the presence of a small focus of intramucosal, grade 1, endometrioid-type adenocarcinoma. Immunohistochemically, the tumor cells were positive for estrogen and progesterone receptors. CONCLUSION: We concluded that while MPA treatment had been effective, it had not completely eradicated the carcinomatous lesion, which remained during and after the term pregnancy.

Adenocarcinoma↗

PubMed2000

Relation between thymidylate synthase expression and survival in colon carcinoma, and determination of appropriate application of 5-fluorouracil by immunohistochemical method.

BACKGROUND: Thymidylate synthase (TS) is regarded as a parameter of 5-fluorouracil (5-FU) chemosensitivity for colorectal carcinoma. Recent researchers indicate that the chemosensitivity of 5-FU for colorectal carcinoma with low expression of TS is better than tumors with high expression of TS. But the relation between TS expression and overall survival of curatively resected colorectal cancer patients has been less studied. METHODS: Specimens of curatively resected colon carcinoma from 148 patients were included in this study. TS expression in the tumor was assessed by immunohistochemical staining technique, and the patients were categorized into TS-(+) and TS-(-) groups. First, the relation between TS expression and survival of patients was examined. Next, for each group, we compared survival between the chemotherapy-(+) and the chemotherapy-(-) subgroup. RESULTS: Overall survival was significantly better in the TS-(-) group (n = 107) than in the TS-(+) group (n = 41) (P = .0003). In the TS-(-) group, there was little difference between the chemotherapy-(+) and the chemotherapy-(-) subgroup. In the TS-(+) group, the survival of the chemotherapy-(+) subgroup was significantly better than the chemotherapy-(-) subgroup (P = .0439). CONCLUSIONS: TS, itself, may be a prognostic factor for colon carcinoma; and 5-FU adjuvant chemotherapy may be appropriate for colon carcinoma with high expression of TS.

Antimetabolites, Antineoplastic↗

PubMed2000

Significance of alpha 9 beta 1 and alpha v beta 6 integrin expression in breast carcinoma.

BACKGROUND: Both alpha 9 beta 1 and alpha v beta 6 integrins have been newly identified from the tracheal epithelium of guinea pig. It has been pointed out that alpha 9 beta 1 functions as a receptor for tenascin-C and osteopontin. As for the ligands of alpha v beta 6, fibronectin and tenascin-C have been identified. It has not been ascertained whether alpha 9 beta 1 and alpha v beta 6 are expressed in normal breast tissue, benign breast lesion or breast carcinoma. METHODS: Immunohistochemical staining for alpha 9 beta 1 and alpha v beta 6 was performed in benign breast lesion and breast carcinoma specimens. Western blotting was carried out on 11 breast carcinoma cases. RESULTS: alpha 9 beta 1 was expressed in the cytoplasm of carcinoma cells in 23 of 90 cases (26%) and alpha v beta 6 in the membrane of carcinoma cells in 16 of 90 cases (18%). However, these findings of alpha 9 beta 1 and alpha v beta 6 did not correlate with any clinicopathological factors including the patients' age, tumor size, histological type of carcinoma, location of carcinoma cells and hormone receptor status. With regard to the histological grade of carcinoma, alpha v beta 6 and alpha 9 beta 1 expression did not statistically correlate, although no expression of alpha v beta 6 was observed in 14 cases of Grade I. On Western-blott analysis strong and weak bands consistent with alpha v beta 6 were noted in the membrane fraction extracted from breast carcinoma cells. On the other hand weak bands consistent with alpha 9 subunit were noted in the whole cell lysates of breast carcinoma cells and very weak or no bands consistent with alpha 9 subunit were noted in the membrane fraction extracted from the breast carcinoma cells. CONCLUSIONS: Significance of alpha 9 beta 1 and alpha v beta 6 integrins expression in breast carcinoma was still unknown on clinicopathological examination. The findings of Western blot analysis may indicate that the transportation system of glycoproteins such as integrins to the cell membrane of carcinoma cells is disturbed, although these integrins can be produced.

Adenoma↗

PubMed2000

Adnexal torsion: MR imaging findings of viable ovary.

We report a case of torsed ovarian cystic tumor, in which contrast-enhanced high-resolution MR images accurately contributed to the diagnosis of torsion despite the lack of symptoms and to the preoperative evaluation of viability of the edematous ovary. Accurate preoperative assessment by MR images and prompt conservative surgical approach succeeded in salvaging the involved ovary.

Adult↗

PubMed2000

Secondary acute nonlymphocytic leukemia following successful chemotherapy combining cisplatin, doxorubicin, and cyclophosphamide for stage IV epithelial ovarian cancer.

Although chemotherapy is indispensable for the treatment of ovarian cancer, secondary acute leukemia has become increasingly important as one of the most unfavorable late effects according to widespread long-term chemotherapy. We report a patient suffering from acute nonlymphocytic leukemia (ANLL) 3 years after treatment for stage IV ovarian cancer began.

Antibiotics, Antineoplastic↗

PubMed2000

Detection of intracellular nitric oxide using a combination of aldehyde fixatives with 4,5-diaminofluorescein diacetate.

Using 4,5-diaminofluorescein diacetate (DAF-2DA), which was recently developed for the detection of intracellular nitric oxide (NO) in living cells, we examined the sensitivity of intracellular NO in cells treated with some fixatives. Cultured human umbilical vein endothelial cells loaded with DAF-2DA in the presence of 10(-6) M acetylcholine showed intense fluorescence when fixed in paraformaldehyde or glutaraldehyde, but no fluorescence could be detected after fixation in ethanol or acetone. Fluorescence generation depended on the combination of each aldehyde fixative with DAF-2, which is produced enzymatically from DAF-2DA within the cells. Subtracting the fluorescence intensity of non-activated controls from that of cells activated by acetylcholine indicated the NO produced in the stimulated cells, since the control cells that took up DAF-2DA also generated fluorescence when treated with aldehyde fixatives. Thus, detection of intracellular NO by combining aldehyde fixatives with DAF-2DA is useful for examining the functions of NO in cells both in situ and in vivo.

Acetone↗

PubMed2000

Near-infrared monitoring of myocardial oxygenation during ischemic preconditioning.

BACKGROUND: Ischemic preconditioning has been advocated as a method of cardioprotection for minimally invasive direct coronary artery bypass. This study was performed to estimate the cardioprotective effect of ischemic preconditioning before ischemia by examining the changes in myocardial tissue oxygenation and also to examine whether adenosine triphosphate-sensitive potassium channel opener enhances the cardioprotective effect of ischemic preconditioning. METHODS: Myocardial ischemia was induced in three groups of 6 dogs by temporary occlusion of the left anterior descending coronary artery. Group 1 dogs received a 30-minute coronary occlusion and subsequent 3-hour reperfusion. Groups 2 and 3 dogs underwent three periods of 5-minute coronary occlusion and 5-minute reperfusion and then received 30-minute sustained ischemia and 3-hour reperfusion. In group 3, nicorandil was administered during the procedure. Myocardial oxygenation was measured using three-wavelength near-infrared spectroscopy. Myocardial blood flow was measured by the colored microsphere method. RESULTS: During ischemic preconditioning the myocardial tissue oxygen saturation decreased rapidly at coronary occlusion and increased at reperfusion. It was increased stepwise at the second and third coronary occlusion. Myocardial oxygen saturation during 30-minute sustained ischemia was significantly higher in groups 2 and 3 than in group 1 (p < 0.05). The myocardial tissue hemoglobin concentration showed similar changes to myocardial oxygen saturation. During 30-minute sustained ischemia, it was significantly higher in group 2 than in group 1 (p < 0.001), and it was significantly higher in group 3 than in groups 1 and 2 (p < 0.05). Regional myocardial blood flow showed no difference after 30 minutes of sustained ischemia among the three groups. Troponin-T levels were significantly lower in groups 2 and 3 than in group 1 (p < 0.01). CONCLUSIONS: Ischemic preconditioning had beneficial effects on myocardial oxygenation during sustained ischemia, and the protected state of the myocardium could be monitored with the use of near-infrared spectroscopy. Ischemic preconditioning coupled with nicorandil administration might provide protection for minimally invasive direct coronary bypass.

Animals↗

PubMed2000

Therapeutic angiogenesis with intramyocardial administration of basic fibroblast growth factor.

BACKGROUND: Basic fibroblast growth factor (bFGF) induces endothelial cell and smooth muscle cell proliferation and stimulates angiogenesis. This study was designed to evaluate the effects of intramyocardial administration of bFGF on myocardial blood flow, angiogenesis, and ventricular function in a canine acute infarction model. METHODS: Myocardial infarction was induced in 12 dogs by ligation of the left anterior descending coronary artery. Within 5 minutes after coronary occlusion, 100 microg of human recombinant bFGF in 1 mL of saline was injected into the infarct and border zone in 6 dogs, whereas saline alone was used in 6 control dogs. Myocardial blood flow was determined with colored microspheres before and immediately after coronary ligation and again 3, 7, 14, and 28 days after treatment and it was expressed as percent of normal. Angiogenesis was evaluated by immunohistochemical studies 28 days later. Cardiac function was evaluated by repeated echocardiographic measurement. RESULTS: Treatment with bFGF significantly increased the endocardial blood flow in the border zone (7 days after infarction, 75%+/-7% and 41% +/-7% in the bFGF and control groups, respectively, p<0.01) as well as epicardial blood flow in the infarcted zone. Treatment with bFGF significantly increased the capillary density (39.7+/-2.3 and 22.7+/-1.1 vessels per visual field in the bFGF and control groups, respectively, p<0.01) as well as arteriolar density in the border zone. Treatment with bFGF significantly reduced the change in ratio of thickness of the infarcted wall to the normal wall (44%+/-6% and 26% +/-5% in the bFGF and control groups, respectively, p<0.05). It improved the left ventricular ejection fraction (7 days after infarction, 0.54+/-0.02 and 0.37+/-0.03 in the bFGF and control groups, respectively, p<0.01). CONCLUSIONS: Intramyocardial administration of bFGF increased the regional myocardial blood flow, reduced thinning of the infarcted region, and improved ventricular function in acute myocardial infarction. Intramyocardial administration of bFGF may be a new therapeutic approach for patients with acute myocardial infarction.

Animals↗