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Y Obata

Publications and source records attributed to Y Obata.

At least 55 records · Page 3Linked to original sources
PubMed1998

Enzymatic amplification and expression of bovine interleukin-1 receptor antagonist cDNA.

cDNA generated from lipopolysaccharide-stimulated bovine peripheral blood mononuclear cells was used to amplify and clone the bovine interleukin-1 receptor antagonist (IL-1ra) using primers derived from semi-conserved regions between human and mouse IL-1ra sequences. 5' and 3' terminal sequences of bovine IL-1ra were amplified by 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of bovine IL-1ra demonstrated 80%, 78%, 78%, 77% and 76% homology with human, mouse, rat, rabbit and equine sequences, respectively. Recombinant bovine IL-1ra produced in Escherichia coli suppressed the growth inhibitory activity of bovine IL-1beta on A375 cells in a dose-dependent manner, indicating that the present bovine IL-1ra cDNA encodes biologically active proteins.

Amino Acid Sequence↗
PubMed1998

A paired wedge filter system for compensation in dose differences.

OBJECTIVE: In radiotherapy, it is important to conform the high dose volume to the planned target volume. A variable thickness paired wedge filter system was developed to compensate for dose inhomogeneity arising from field width segment variation in conformal irradiation. MATERIALS AND METHODS: The present study used a 6 MV linear accelerator equipped with multileaf collimator leaves and a paired wedge compensating filter system. The dose variation due to field width was measured in each field segment width. The variation in attenuation of the compensators was measured as a function of filter position. As the field width increases, the relative absorbed dose also increases; this is the point of requiring compensation, so it can be in reverse proportion. RESULTS: As the field width increases, the relative absorbed dose also increases; this is why compensation is required and thus it must be in reverse proportion. Attenuation of the absorbed dose by the paired filters was in proportion to the filter position. The filter position to compensate for the difference of absorbed doses was defined by the square root of the field width. For a field varying in width from 4 to 16 cm, the variation in the absorbed dose across the field was reduced from 12% to 2.7%. CONCLUSION: This paired wedge filter system reduced absorbed dose variations across multileaf collimator shaped fields and can facilitate treatment planning in conformal therapy.

Humans↗
PubMed1998

Disruption of primary imprinting during oocyte growth leads to the modified expression of imprinted genes during embryogenesis.

Parthenogenetic embryos, which contained one genome from a neonate-derived non-growing oocyte and the other from a fully grown oocyte, developed to day 13.5 of gestation in mice, 3 days longer than previously recorded for parthenogenetic development. To investigate the hypothesis that disruption of primary imprinting during oocyte growth leads to the modified expression of imprinted genes and this parthenogenetic phenotype, we have examined Peg1/Mest, Igf2, Peg3, Snrpn, H19, Igf2r and excess p57KIP2. We show that paternally expressed genes, Peg1/Mest, Peg3 and Snrpn, are expressed in the parthenotes, presumably due to a lack of maternal epigenetic modifications during oocyte growth. In contrast, the expression of Igf2, which is repressed in a competitive manner by transcription of the H19 gene, was very low. Furthermore, we show that the maternally expressed Igf2r and p57KIP2 genes were repressed in the alleles of the non-growing oocyte indicating maternal modifications during oocyte growth are necessary for its expression. Thus, our results show that primary imprinting during oocyte growth exhibits a crucial effect on both the expression and repression of maternal alleles during embryogenesis.

Animals↗
PubMed1998

Protective effect of passive immunization against TNF-alpha in mice infected with Sendai virus.

TNF-alpha has been reported to be induced in mice infected with Sendai virus. We evaluated the role of TNF-alpha in the virus infection. TNF-alpha was induced locally in proportion to virus titers in the lung. The activity was correlated with suppression of body weight gain. Passive immunization against TNF-alpha improved body weight gain and ameliorated pneumonic lesions in infected mice, and prevented them from lethal infection, but lung virus induced emaciation, pneumonic lesions and death were mediated by TNF-alpha.

Animals↗
PubMed1998

Effect of cod-liver oil extract on the buccal permeation of ergotamine tartrate.

Ergotamine tartrate (ET) is used clinically in the treatment of migraines. However, the bioavailability of ET is rather poor following oral administration. Therefore, we tried to improve ET delivery using buccal administration. The purpose of this study was to investigate the characteristics of the permeation of ET through the hamster cheek pouch in vitro using a two-chamber diffusion cell, and to evaluate the effect of permeation enhancers on the transbuccal delivery of ET. Cod-liver oil extract (CLOE), polyoxyethylene hydrogenated castor oil (HCO 60), sodium glycocholate (GC), and sodium caprate (CA) were selected as premeation enhancers considering their low irritancy of the mucosa. When the enhancers were added to the donor cell at a 5% concentration each, the ET permeation rate markedly increased compared with that in a control not containing enhancer. Among these enhancers, CLOE exhibited the greatest effect. Because CLOE is composed of 16 kinds of fatty acids, the enhancement action of each of the major components was separately determined. As major fatty acids, palmitic acid, oleic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) were selected and their enhancing effects were studied. The enhancing effect of each fatty acid was significantly lower than that of CLOE.

Animals↗
PubMed1997

Requirement of CD4 T cells for skin graft rejection against thymus leukemia (TL) antigen and multiple epitopes on the TL molecule recognized by CD4 T cells.

By selective depletion of CD4 and CD8 T cells in vivo using the respective mAbs, we demonstrate that CD4 T cells are necessary for skin graft rejection against thymus leukemia (TL) Ag. The skin expressing T3b-TL Ag from transgenic C3H Tg.Con.3-1 mice given chimeric H-2Kb/T3b-TL gene was rejected when grafted onto C3H/He recipient mice. Depletion of CD4, but not of CD8, T cells blocked rejection. CD8 CTL were generated in MEM (control)-treated C3H/He recipient mice, while Thy-1+ CD4- CD8- CTL were generated in CD8-depleted recipient mice after rejection. However, no CTL were generated in CD4-depleted or both CD4- and CD8-depleted recipient mice. Thus, the generation of both CD8 and Thy-1+ CD4- CD8- CTL was dependent on CD4 T cells. Ab blocking indicated that both CD8 and Thy-1+ CD4- CD8- CTL were TCR alphabeta and recognized TL Ag. We furthermore demonstrated that CD4 T cells in spleen cells from C3H/He mice that had rejected C3H Tg.Con.3-1 skin showed a weak, but significant, proliferative response to in vitro stimulation with mitomycin C-treated C3H Tg.Con.3-1 spleen cells. Analysis of the reactivity of bulk CD4 T cell lines to 73 synthetic overlapping peptides encompassing the entire T3b-TL molecule showed that CD4 T cells recognized multiple epitopes on the T3b-TL molecule in an APC-dependent manner.

Amino Acid Sequence↗
PubMed1997

NF-kappa B RelA-deficient lymphocytes: normal development of T cells and B cells, impaired production of IgA and IgG1 and reduced proliferative responses.

To investigate the function of NF-kappa B RelA (p65), we generated mice deficient in this NF-kappa B family member by homologous recombination. Mice lacking RelA showed liver degeneration and died around embryonic day 14.5. To elucidate the role of RelA in lymphocyte development and function, we transplanted fetal liver cells of 13.5-day embryos from heterozygote matings into irradiated SCID mice. Within 4 weeks, both T and B cells had developed in the SCID mice receiving relA-/- fetal liver transplants, similar to the relA+/+ and +/- cases. T cells were found to mature to Thy-1+/TCR alpha beta +/CD3+/CD4+ or CD8+, while B cells had the ability to differentiate to IgM+/B220+ and to secrete immunoglobulins. However, the secretion of IgG1 and IgA was reduced in RelA-deficient B cells. Furthermore, both T and B cells lacking RelA showed marked reduction in proliferative responses to stimulation with Con A, anti-CD3, anti-CD3 + anti-CD28, LPS, anti-IgM, and PMA + calcium ionophore. The results indicate that RelA plays a critical role in production of specific Ig isotypes and also in signal transduction pathways for lymphocyte proliferation.

Animals↗
PubMed1997

Synthesis and biological evaluation of orally active matrix metalloproteinase inhibitors.

The synthesis and biological evaluation of orally active inhibitors of matrix metalloproteinase are reported. Modifications of the P2' position and the alpha-substituent of hydroxamic acid derivatives were carried out, and revealed that the P2' substituent influenced the MMP inhibitory activities in vitro and in plasma after oral administration. The hydroxamates with phenylglycine at the P2' position were absorbed well orally. Compound 15e, which exhibited the longest duration of inhibitory activity in plasma after oral administration among the phenylglycine derivatives (5a-5d, 15a, 15c, 15e), was evaluated in a rat adjuvant arthritis model. A reduction in hind foot pad swelling and improvements of some inflammatory parameters were demonstrated when the compound was administered orally. These results indicate the potential of MMP inhibitors for rheumatoid arthritis.

Absorption↗
PubMed1997

Detection of interleukin-1 beta in sera and colostrum of dairy cattle and in sera of neonates.

In order to obtain basic information about bovine interleukin-1 (IL-1 beta), levels of IL-1 beta in sera and milk of clinically normal mature Holstein cattle before and after parturition and in sera of newborn calves were examined by ELISA. The level of IL-1 beta was undetectable in sera of mature cattle around the time of artificial insemination, but the concentration gradually increased and reached a peak at parturition and then decreased again to an undetectable level. IL-1 beta in milk was detected on the day of parturition but not thereafter. IL-1 beta mRNA was detected by reverse transcription-polymerase chain reaction in the cells from milk collected during 20 days before and 2 to 3 days after parturition, but was not detected thereafter. Although IL-1 beta was not detected in all the sera of newborn calves, the concentration transiently increased with peak titers on day 3 and became undetectable by day 14 after birth. Newborns that showed serum IL-1 beta on day 3 had been fed on colostrum in which the IL-1 beta concentration was significantly higher than that in colostrum that had been fed to newborns having no detectable IL-1 beta on day 3. These results indicate that IL-1 beta is induced in association with pregnancy in healthy dairy cattle and that the cytokine might be transferred to neonates via colostrum.

Amino Acid Sequence↗
PubMed1997

Breast ultrasonography: computer-aided diagnosis using fuzzy inference.

On the basis of detailed analysis of ultrasonographic features in 105 breast masses (55 malignant, 50 benign), a computer-aided diagnostic system using fuzzy inference has been developed. Ultrasonographic features of a mass for the input data included shape, border, halo (boundary echoes), internal echoes, posterior echoes, and edge shadows (bilateral shadows). The probability of malignancy was described by an actual number ranging from 0.0 to 1.0. The algorithm of inference was constructed, and a sensitivity of 94.5% and specificity of 76.0% for cancer diagnosis were obtained.

Adolescent↗
PubMed1997

[Transesophageal echocardiographic findings of cortical and perforating infarctions in patients with atrial fibrillation].

The predictive value of transesophageal echocardiography was investigated for the risk stratification of atherothrombotic or embolic cerebral infarction in patients with atrial fibrillation. Left atrial spontaneous echo contrast, peak flow velocity in the left atrial appendage and generalized atherosclerosis as estimated by the intima-media wall thickness of the thoracic aorta were assessed by transesophageal echocardiography in consecutive patients with paroxysmal (n = 25) or chronic (n = 60) atrial fibrillation (mean [+/-SD] age; 63 +/- 11 years). All patients underwent brain computed tomography or magnetic resonance imaging to evaluate the presence or absence of cerebral infarction. The location of cerebral infarction was divided into two territories, the cortical branch (cortical infarction) and deep perforators (perforating infarction), to evaluate embolic and atherothrombotic cerebral infarction, respectively. Cortical and perforating infarctions were found in 42% and 16% of all patients, respectively. The grade of spontaneous echo contrast was higher in patients with cortical infarction than in those without cortical infarction. Patients with perforating infarction showed thicker aortic wall compared with patients without perforating infarction. Other parameters had no predictive value to differentiate perforating from cortical infarctions. Multiple regression analysis revealed that spontaneous echo contrast and age were independent predictors of embolic cortical infarction, whereas intima-media wall thickness of the aorta and hypertension were useful in predicting the risk of atherothrombotic perforating infarction. Transesophageal echocardiography is useful for predicting embolic cortical infarction and atherothrombotic perforating infarction.

Atrial Fibrillation↗
PubMed1997

[Method for determining optimal intracavitary radiotherapy conditions for carcinoma of the uterine cervix with tumor infiltration of the vaginal wall].

Stage III carcinoma of the uterine cervix is occasionally accompanied by tumor infiltration of the vaginal wall. Currently, the vaginal wall has to be irradiated in the same manner as the uterine cervix. The authors have developed a system for determining the optimal irradiation conditions for treating the two regions, uterine cervix and vaginal wall, at the same time. A comparison of two methods is shown in simulation, and then a clinical case is reported. The first method consists of two treatment plans, one for the uterine cervix without tumor infiltration of the vaginal wall, and the other for the vaginal wall without carcinoma of the uterine cervix. The second, newly developed method considers the two regions together. Irradiation times of ovoid sources obtained with the second method are 15-25% less than those of the first method. Isodose curves obtained with the two methods are very different, and thus the uterine cervix and vaginal wall must be considered together in order to determine irradiation conditions.

Brachytherapy↗
PubMed1996

Positive selection of gamma delta CTL by TL antigen expressed in the thymus.

To elucidate the funciton of the mouse TL antigen in the thymus, we have derived two TL transgenic mouse strains by introducing Tl alpha 2-3 of A strain origin with its own promoter onto a C3H background with no expression of TL in the thymus. These transgenic mouse strains, both of which express high levels of Tla2-3-TL antigen in their thymus, were analyzed for their T cell function with emphasis on cytotoxic T lymphocyte (CTL) generation. A T cell response against TL was induced in Tg. Tla2-3-1, Tg. Tla2-3-2, and control C3H mice by skin grafts from H-2Kb/T3b transgenic mice, Tg.Con.3-1, expressing T3b-TL ubiquitously. Spleen cells from mice that had rejected the T3b-TL positive skin grafts were restimulated in vitro with Tg. Con.3-1 irradiated spleen cells. In mixed lymphocyte cultures (MLC), approximately 20% and 15% of Thy-1+ T cells derived from Tg.Tla2-3-1 and Tg.Tla2-3-2, respectively, expressed TCR gamma delta, whereas almost all those from C3H expressed TCR alpha beta. The MLC from Tg. Tla2-3-2 and C3H demonstrated high CTL activity against TL, while those from Tg. Tla2-3-1 had little or none. The generation of gamma delta CTL recognizing TL in Tg. Tla2-3-2, but not C3H mice, was confirmed by the establishment of CTL clones. A total of 14 gamma delta CTL clones were established from Tg. Tla2-3-2, whereas none were obtained from C3H. Of the 14 gamma delta CTL clones, 8 were CD8+ and 6 were CD4-CD8- double negative. The CTL activity of all these clones was TL specific and inhibited by anti-TL, but not by anti-H-2 antibodies, demonstrating that they recognize TL directly without antigen presentation by H-2. The CTL activity was blocked by antibodies to TCR gamma delta and CD3, and also by antibodies to CD 8 alpha and CD8 beta in CD8+ clones, showing that the activity was mediated by TCR gamma delta and coreceptors. The thymic origin of these gamma delta CTL clones was indicated by the expression of Thy-1 and Ly-1 (CD5), and also CD8 alpha beta heterodimers in CD8+ clones on their surfaces and by the usage of TCR V gamma 4 chains in 12 of the 14 clones. Taken together, these results suggest that Tla2-3-TL antigen expressed in the thymus engages in positive selection of a sizable population of gamma delta T cells.

Amino Acid Sequence↗
PubMed1996

Inhibition by CsA and FK506 of the in vitro proliferative response of gamma delta T cells on stimulation with anti-TCR delta monoclonal antibody.

Tg.Tla'-3-1 mice are transgenic C3H/He mice with a Tlaa-3 gene derived from A mice. We demonstrated that spleen cells from Tg.Tlaa-3-1 mice, but not C3H/He mice, showed a proliferative response on stimulation with immobilized anti-TCR delta monoclonal antibody (mAb 3A10). Thus, Tg.Tlaa-3-1 mice can be useful for analysis of the function of gamma delta T cells. Using spleen cells from Tg.Tlaa-3-1 mice, we showed that cyclosporin A (CsA) and FK506 inhibited the in vitro proliferative response of gamma delta T cells on stimulation with anti-TCR delta mAb. The dose-dependent inhibitory effect of CsA and FK506 on proliferation of gamma delta T cells on stimulation with anti-TCR delta mAb was similar to that on proliferation of alpha beta T cells on stimulation with anti-TCF beta mAb. Inhibition by CsA and FK506 of proliferation of gamma delta T cells was not reversed by addition of rIL 2 (recombinant interleukin 2) during culture. The proliferative response of gamma delta T cells among spleen cells from TCR alpha beta-depleted Tg.Tlaa-3-1 mice was also inhibited by CsA and FK506, suggesting that the inhibition directly affected gamma delta T cells without mediation by alpha beta T cells.

Animals↗
PubMed1996

Monoclonal antibody against the fusion junction of a deletion-mutant epidermal growth factor receptor.

A mouse monoclonal antibody (IgG2b), 3C10, was produced against the truncated epidermal growth factor receptor (EGFR), encoded by the (type III) in-frame deletion mutation of 801 nucleotides of EGFR affecting the external domain, known to be expressed in some human glioblastoma. As this mutation newly generates a glycine residue at the fusion point, a 14 amino acid peptide around the fusion junction including this glycine was chemically synthesised and used for immunisation of (B6 x DBA/2) F1 mice. Flow cytometric analysis showed 3C10 antibody staining of a mouse NIH/3T3 transfectant (ERM5) with the type III EGFR deletion-mutant gene, but not one with wild-type EGFR. The antibody immunoprecipitated the truncated EGFR protein with a molecular mass of approximately 140 kDa from ERM5 cells. Immunostaining of glioblastomas revealed binding in the case with the type III EGFR mutation, the five other specimens without the mutation being negative despite overexpression of EGFR in some cases.

3T3 Cells↗
PubMed1996

Epigenetic modifications during oocyte growth correlates with extended parthenogenetic development in the mouse.

In mammals, the maternal and paternal genomes are required for embryonic development. This is due to genomic imprinting which leads to the expression or repression of genes solely on the basis of the parent from which they were inherited. As a result, parthenogenetic embryos die before day 10 of gestation and show limited development of extra-embryonic membranes. Maternal imprinting is established during oogenesis and is associated with allele specific modifications in DNA methylation. We have investigated epigenetic modifications during oocyte growth using nuclear transfer techniques to produce mature oocytes with maternal chromatin derived from non-growing oocytes. Parthenogenetic activation of such oocytes leads to the development of normal size fetuses with a well developed placenta on day 13.5 of gestation; three days further than previously recorded for parthenogenetic development. In contrast, after fertilization, only one embryo was recovered on 9.5 days of gestation. Further, in these embryos we investigated the well characterized methylation pattern of the maternally expressed insulin-like growth factor II receptor gene (Igf2r) and found that the pattern of methylation was indeed different to that of fertilized control embryos. Thus, the embryonic phenotypes observed here correlate with changes in epigenetic events that normally occur during oocyte growth.

Animals↗
PubMed1996

Inhibitory effects of the new anti-platelet agent KBT-3022 and its metabolite on rabbit neutrophil function in vitro.

The effects of the new anti-platelet agent KBT-3022, ethyl 2-[4,5-bis(4-methoxyphenyl)-thiazol-2-yl]pyrrol-1-ylacetate, and its metabolite desethyl KBT-3022 on rabbit neutrophil function were investigated in comparison with the effects of acetylsalicylic acid (ASA), ticlopidine hydrochloride (TP), cilostazol (CIL) and indomethacin (IM). The adhesion and migration of neutrophils induced by formyl-methionyl-leucyl-phenylalanine (fMLP) were inhibited by all the compounds tested, their rank order of potency being KBT-3022 = desethyl KBT-3022 > TP = CIL = IM > ASA. KBT-3022, desethyl KBT-3022, CIL and IM all suppressed fMLP-induced increases in the intracellular free Ca2+ concentration ([Ca2+]i) in neutrophils, their potencies correlating with their inhibitory effects on fMLP-induced adhesion and migration. KBT-3022 (1 microM), desethyl KBT-3022 (1-10 microM) and CIL (10 microM) but not IM significantly inhibited both neutrophil migration and the increase in [Ca2+]i induced by leukotriene B4 (LTB4). KBT-3022 (1 microM) and desethyl KBT-3022 (1 microM) suppressed the increase in [Ca2+]i induced by complement C5a. Although KBT-3022 and desethyl KBT-3022 did not influence [3H]LTB4 and [125I]C5a specific binding, [3H]fMLP specific binding was inhibited by desethyl KBT-3022 (IC50: 1.9 microM). Neutrophil adhesion and superoxide anion production stimulated by phorbol 12-myristate 13-acetate were partially inhibited by KBT-3022 (1 microM) and desethyl KBT-3022 (1-10 microM). These results suggest that KBT-3022 and desethyl KBT-3022 have a wider spectrum of action and are more potent inhibitors of neutrophil activation than ASA, TP, CIL and IM.

Animals↗