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Biomedical subjects

Y Inoue

Publications and source records attributed to Y Inoue.

At least 793 records · Page 44Linked to original sources
PubMed1997

Structural organization of the human vitamin D receptor chromosomal gene and its promoter.

The vitamin D receptor (VDR) is known to mediate the pleiotropic biological actions of 1,25-dihydroxyvitamin D3 through its ability to modulate the expression of target genes. The regulation of this ligand-activated cellular transcription factor is reported to occur at both transcriptional and posttranslational levels. To begin to address the molecular basis by which the VDR gene is regulated transcriptionally, we report here an initial characterization of the human VDR gene and its promoter. We isolated several overlapping A-phage and cosmid clones that cover more than 100 kb of human DNA and contained the entire VDR gene. The gene is comprised of 11 exons that, together with intervening introns, span approximately 75 kb. The noncoding 5'-end of the gene includes exons 1A, 1B, and 1C. Eight additional exons (exons 2-9) encode the structural portion of the VDR gene product. While primer extension and S1 nuclease-mapping studies reveal several common transcriptional start sites, three unique mRNA species are produced as a result of the differential splicing of exons 1B and 1C. The DNA sequence lying upstream of exon 1A is GC rich and does not contain an apparent TATA box. Several potential binding sites for the transcription factor SP1 and other activators are evident. Fusion of DNA fragments containing putative promoter sequences upstream of the luciferase structural gene followed by transient transfection of these plasmids into several mammalian cell lines resulted in significant reporter activity. Due to the size and complexity of the 5'-end of the VDR gene, we examined the activity of a DNA fragment surrounding exon 1C. An intron fragment 3' of exon 1C conferred retinoic acid responsivity when fused to a reporter gene plasmid, suggesting a molecular mechanism for the previously observed ability of retinoic acid to induce the VDR. The recovery of the gene for the human VDR will enable further studies on the transcriptional regulation of this gene.

Bacteriophage lambda↗
PubMed1997

Oxidative stability of docosahexaenoic acid-containing oils in the form of phospholipids, triacylglycerols, and ethyl esters.

The peroxidative stability of docosahexaenoic acid (DHA)-containing oils (DHA at 10.7 mol% of the total fatty acids), in the form of phospholipids (PL), triacylglycerols (TG), and ethyl esters (EE) with the same constituent fatty acids, was investigated in the dark at 25 degrees C in a bulk phase, and compared with that of control palm oil (supplemented with 20% soybean oil). The oxygen absorption of the DHA-containing oil was significantly lower in the form of PL than in the form of TG and EE during a 10-week oxidation, and the oxygen absorption of PL was almost equivalent to that of the control oil. A gas chromatographic analysis showed that 90% of initial DHA was retained in the form of PL after the 10-week oxidation, while TG and EE respectively more rapidly decayed with the loss of 97% and 64% of DHA. Tocopherol in the form of TG and EE had also completely decayed after the oxidation, while 37% of the initial tocopherol remained in the form of PL. The peroxide and carbonyl values of TG and EE showed large increases after the oxidation, but no such increase was observed for PL. These results show that DHA-containing oil in the form of PL was more resistant to the oxidative degradation of DHA than that in the form of TG and EE in a bulk phase.

Darkness↗
PubMed1997

Variation from cytopathogenic biotype to non-cytopathogenic biotype is correlated with the deletion of cellular sequence from bovine viral diarrhea viruses.

Non-cytopathogenic (NCP) viruses of bovine viral diarrhea (BVD) virus were detected at a low ratio by the reverse plaque formation method from virus samples after several plaque clonings of cytopathogenic (CP) BVD viruses; NADL and Osloss strains. This phenomenon suggests that the NCP BVD viruses are produced at a low ratio during the propagation of CP BVD viruses in vitro. To investigate the differences between the parent CP BVD virus and the NCP BVD virus as a real progeny, the regions flanking the insertion of cellular mRNA in the p125 domain of NADL and Osloss strains were amplified by RT-PCR and cloned into pGEM 3Z plasmid vector, and then sequenced. Consequently, it was confirmed that sequences of cellular mRNA insertion of CP BVD viruses, NADL and Osloss strains, were completely and exactly deleted from the NCP BVD viruses which were real progeny of CP BVD viruses, NADL and Osloss strains. These results suggest that NCP BVD viruses may revert from CP biotype to NCP biotype by the deletion of cellular mRNA insertion in the viral genome of CP BVD viruses (NADL and Osloss strains).

Amino Acid Sequence↗
PubMed1997

Comparison of cellular characteristics between human hepatoma cell lines with wild-type p53 and those with mutant-type p53 gene.

Characteristics of human hepatoma cell lines with the wild-type p53 were compared with those of human hepatoma cell lines with the mutant-type p53. The p21 protein located downstream of p53 was expressed in cell lines with the wild-type p53 but was not expressed in cell lines with the mutant-type p53. As to other tumor suppressor genes such as p16 and p27, there was no difference in their expression between both types of cell lines. In addition, no marked difference was observed in the activities of CDK2 and CDK4 between cell lines with the wild-type and the mutant-type p53. Phosphorylated Rb protein was detected in all cell lines except the HLE line, indicating that this cell line may have a deletion of and/or a mutation of the Rb gene. These results indicate that abnormalities of tumor suppressor genes other than p53, p16, p27, and Rb may be involved in hepatocarcinogenesis. The population doubling time of the wild-type p53 cells was significantly longer than that of the mutant p53 cells. Neither type of cell line showed a specific chromosome distribution which would indicate karyotype instability. The cell lines expressing the wild-type p53 produced tumors at lower frequency than those with the mutant p53 gene. Although there was no significant difference in effects of TGF-beta 1, EGF, cholera toxin, and db-cAMP on cell growth between the two types of cells, all three cell lines with the wild-type p53 were resistant to cytotoxicity of TNF-alpha, while two of the three with the mutant p53 were very sensitive to its cytotoxic effects.

Animals↗
PubMed1997

Yeast functional assay of the p53 gene status in human cell lines maintained in our laboratory.

We used a yeast functional assay (functional analysis of separated alleles in yeast: FASAY) to determine the p53 gene status of human cell lines maintained in our laboratory. This assay enables the researcher to score wild-type p53 expression on the basis of the ability of expressed p53 to transactivate the reporter gene HIS 3 via the p53-responsive GAL 1 promoter in Saccharomyces cerevisiae. The cell lines examined were ten hepatoma, two hepatoblastoma, three in vitro immortalized fibroblast, two osteosarcoma, a chondrosarcoma, an ovarian teratocarcinoma and a colon cancer cell line. Out of 20 cell lines, 11 cell lines had mutations in both alleles of the p53 gene, and another 8 cell lines had no mutation in the p53 gene. Thus, 55% of the cell lines examined had mutations in the p53. Interestingly, PA-1 cells had both the normal and the mutant p53 alleles, showing that FASAY is a useful method for detecting the wild-type and mutated p53 genes simultaneously. As for the three liver cell lines harboring HBsAg, there was no relationship between their p53 gene status and the presence of HBsAg. Two cell lines were normal for p53 status, while the other had a mutation of the p53 gene.

Alleles↗
PubMed1997

Severe thrombocytopenia after cytomegalovirus infection in an immunocompetent host--correlation between CMV infection and platelet count in immunocompetent host.

We report a case of severe thrombocytopenia after cytomegalovirus mononucleosis and the changes of the platelet count in 13 cases (including documented case) with CMV mononucleosis (4 cases) or Epstein-Barr virus mononucleosis (9 cases), who were admitted to our hospital from 1991 to 1995. A decrease of the platelet count was observed in some patients at diagnosis compared to the following days, suggesting that thrombocytopenia may be induced by CMV as well as EBV, and mild thrombocytopenia may be more frequent than expected after CMV infection in immunocompetent adults, although severe thrombocytopenia is rare.

Adult↗
PubMed1997

Concentration in blood and organs of dogs after high dose 1,1,1-trichloroethane inhalation.

Dogs were exposed to 1% (v/v) (10,000 ppm) vapor of 1,1,1-Trichloroethane (1,1,1-TCE) by inhalation for 3 min repeated four times at 4 hr intervals under continuous anesthesia. Changes in the 1,1,1-TCE concentration in blood with time, as well as distribution of 1,1,1-TCE in the organs and tissues (lungs, liver, kidneys, heart, brain, and fat around the kidneys and on the abdominal wall) upon completion of the four exposures were studied. Concentrations of 1,1,1-TCE in blood showed the highest level immediately after exposure, and fairly decreased in about 30 min after exposure. The half life of 1,1,1-TCE in blood was 4-12 min after exposure. Upon completion of the exposures (3 min inhalations repeated four times), 1,1,1-TCE concentrations per gram wet weight of each organ ranged from 0.1 to 0.5 microgram/g in the lungs, liver, kidneys, heart and brain. On the other hand, the 1,1,1-TCE concentration in fat ranged from 16.9 to 54.6 micrograms/g, greatly exceeding those in other organs.

Adipose Tissue↗
PubMed1997

Pathogenesis of Chiari malformation: a morphometric study of the posterior cranial fossa.

To investigate overcrowding in the posterior cranial fossa as the pathogenesis of adult-type Chiari malformation, the authors studied the morphology of the brainstem and cerebellum within the posterior cranial fossa (neural structures consisting of the midbrain, pons, cerebellum, and medulla oblongata) as well as the base of the skull while taking into consideration their embryological development. Thirty patients with Chiari malformation and 50 normal control subjects were prospectively studied using neuroimaging. To estimate overcrowding, the authors used a "volume ratio" in which volume of the posterior fossa brain (consisting of the midbrain, pons, cerebellum, and medulla oblongata within the posterior cranial fossa) was placed in a ratio with the volume of the posterior fossa cranium encircled by bony and tentorial structures. Compared to the control group, in the Chiari group there was a significantly larger volume ratio, the two occipital enchondral parts (the exocciput and supraocciput) were significantly smaller, and the tentorium was pronouncedly steeper. There was no significant difference in the posterior fossa brain volume or in the axial lengths of the hindbrain (the brainstem and cerebellum). In six patients with basilar invagination the medulla oblongata was herniated, all three occipital enchondral parts (the basiocciput, exocciput, and supraocciput) were significantly smaller than in the control group, and the volume ratio was significantly larger than that in the Chiari group without basilar invagination. These results suggest that in adult-type Chiari malformation an underdeveloped occipital bone, possibly due to underdevelopment of the occipital somite originating from the paraxial mesoderm, induces overcrowding in the posterior cranial fossa, which contains the normally developed hindbrain. Basilar invagination is associated with a more severe downward herniation of the hindbrain due to the more severely underdeveloped occipital enchondrium, which further exacerbates overcrowding of the posterior cranial fossa.

Adolescent↗
PubMed1997

Increased neutrophil respiratory burst in myeloproliferative disorders: selective enhancement of superoxide release triggered by receptor-mediated agonists and low responsiveness to in vitro cytokine stimulation.

The neutrophil superoxide (O2-)-producing capacity in 57 patients with chronic myeloproliferative disorders (MPDs) and eight patients with chronic myelomonocytic leukemia (CMML) was investigated. O2- release in neutrophils stimulated by chemotactic peptide was markedly increased in all types of chronic MPD, including chronic myelogenous leukemia in both chronic phase and blastic crisis, polycythemia vera, and essential thrombocythemia, but was normal in CMML, which is thought to be a myelodysplastic disorder rather than MPD. Increase in O2(-)-producing capacity in MPD was also observed when other receptor-mediated agonists such as interleukin-8 and concanavalin A were used, but not when phorbol ester, a direct activator of protein kinase C, was used as the triggering agonist of O2- release. Priming effects of granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF), and tumor necrosis factor (TNF) on chemotactic peptide-induced O2- release was observed in all patients with MPD and CMML, though fold enhancement of priming effects was much less in MPD compared with normal subjects. In addition, the priming effects of TNF were less than those of GM-CSF in 10 cases, whereas the priming effects of TNF were consistently and markedly greater than those of GM-CSF in normal subjects. Tyrosine phosphorylation of 42-kDa protein stimulated by G-CSF, GM-CSF, and TNF was observed in CML neutrophils to be identical to that in normal neutrophils. Present results indicate specific potentiation of the receptor-mediated route of signaling that is linked to the respiratory burst and downregulated responsiveness to cytokines in neutrophils in patients with all types of chronic MPD, suggesting in vivo priming of patient neutrophils via certain mechanism by cytokines or related stimuli in these hematological disorders.

Concanavalin A↗
PubMed1997

[Squamous cell carcinoma and basal cell carcinoma].

Both basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are very common in skin malignancy. Operative therapy is the first choice at the treatment for SCC and BCC in early stage. After excision for large skin tumor, occasionally we use skin flap or musculocutaneous flap or free flap. At the old patient of Stage III and IV SCC and BCC, sometimes reduction surgery is useful at the combination therapy. Radiation as adjuvant therapy is sometimes useful for not only SCC but also BCC. The clinical efficacy of CA (C' A') chemotherapy (CDDP, ADM or CBDCA, EPI-ADM), was evaluated on Stage III, IV SCC and BCC. The response rate was 63% in SCC and 75% in BCC.

Adult↗
PubMed1997

[Effectiveness of continuous pulmonary perfusion during total cardiopulmonary bypass to prevent lung reperfusion injury].

The oxygen free radicals and the interaction between neutrophils and endothelium have been implicated in the pathogenesis of lung injury associated with cardiopulmonary bypass (CPB), and in the setting of total CPB, the ischemia-reperfusion injury has been suspected as the mechanism of lung injury. To prevent this reperfusion induced lung injury, we performed continuous pulmonary perfusion during total CPB. We studied 26 infants less than 1 year of age who underwent patch closure of ventricular septal defect. Intermittent mechanical ventilation (5/min) and continuous perfusion of pulmonary artery (30 ml/kg/min) were performed during total CPB in 7 infants (Group P). Whereas 19 infants underwent ordinary CPB (Group N). PaO2/FiO2 ratio was employed for the predictor of lung injury and was calculated before and after CPB. PaO2/FiO2 ratio decreased from 3 to 12 hours after CPB and then increased by 24 hours after CPB in both groups. The lowest PaO2/FiO2 ratio measured at 12 hours after CPB correlated with age and body weight at operation (Spearman's correlation coefficient, 0.59; p = 0.01 and 0.61; p = 0.009, respectively) and strongly correlated with preoperative Rp/Rs ratio (-0.73; p = 0.003). PaO2/FiO2 ratio, however, did not correlate with duration of CPB and aortic cross clamping, preoperative Pp/Ps and Qp/Qs ratio in group N. PaO2/FiO2 ratio of group P at 3, 6, and 12 hours after CPB were higher than those of group N, although there were no significant difference When analysis was made on the infants with high pulmonary vascular resistance (preoperative Rp/Rs ratio > or = 0.1), PaO2/FiO2 ratio of group P (n = 6) at 3, 6 and 12 hours after CPB were higher than those of group N (n = 11), and the difference was statistically significant at 12 hours after CPB (291.1 +/- 15.5 versus 199.6 +/- 27.0, p = 0.027. These results implicate that young age, low body weight and especially high pulmonary vascular resistance were incremental risk factor of lung injury after CPB and, furthermore, ischemia reperfusion injury can be the initiating factor of lung injury. The results also suggest that continuous pulmonary perfusion during total CPB is an effective mean to prevent lung injury particularly for the infants with high pulmonary vascular resistance.

Cardiopulmonary Bypass↗
PubMed1997

Correlation of anterior chamber-associated immune deviation with suppression of corneal epithelial rejection in mice.

PURPOSE: The authors investigated the effect of anterior chamber corneal (AC) inoculation of genetically graft-identical antigen on T-cell immunity and the suppression of alloepithelial rejection in mice. METHODS: Antigen-specific suppression of delayed-type hypersensitivity (DTH) and suppression transferability were tested in BALB/c mice injected with irradiated allogeneic B10.D2 splenocytes into AC. Other groups of BALB/c mice received irradiated B10.D2, BALB/c, or C3H/He splenocytes in the AC of the right eye. Seven days later, B10.D2 or C3H/He corneal lenticules were grafted at the limbus of the left eye (keratoepithelioplasty). Alloepithelial rejection of each grafted eye was evaluated according to clinical findings. The DTH response of the keratoepithelioplasty recipients against B10.D2 minor antigen was tested at the end of clinical observation (4 months after grafting). Also examined was spleen component transfer from BALB/c mice with AC inoculation of B10.D2 splenocytes to syngeneic acceptors and its effect on suppression of epithelial rejection against B10.D2 antigen. RESULTS: Inoculation of B10.D2 splenocytes into BALB/c AC induced antigen-specific DTH suppression, which suppression was transferable. During the 4-month observation period, AC inoculation of B10.D2 minor antigen significantly enhanced the survival of B10.D2-derived epithelium, but not of C3H/He-derived epithelium, in BALB/c mice. However, AC inoculation of BALB/c or C3H/ He splenocytes did not enhance B10.D2 epithelial survival in BALB/c mice. Incapability of antigen-specific DTH response generation was observed in the BALB/c mice with B10.D2 splenocytes in the right AC and B10.D2-derived epithelium in the left eye. Single transfer of spleen components from BALB/c mice with AC inoculation of B10.D2 splenocytes to syngeneic acceptors only delayed B10.D2 minor antigen-stimulated epithelial rejection, whereas supplementary transfers of the identical spleen components at different time intervals showed more significant effect in rejection delay. CONCLUSIONS: The results showed that AC inoculation of B10.D2 splenocytes in BALB/c mice induced antigen-specific suppression of DTH response, in a phenomenon termed anterior chamber-associated immune deviation (ACAID). It also was shown definitely that ACAID can suppress alloepithelial rejection in a murine keratoepithelioplasty model. Adoptive transfer of splenocytes from ACAID-induced mice merely affords short-term suppression of epithelial rejection, suggesting that an additional mechanism may be involved in ACAID maintenance.

Adoptive Transfer↗
PubMed1997

The antigen-bearing eye and the spleen are indispensable in maintaining anterior chamber-associated immune deviation.

PURPOSE: To investigate the role of the eye and the spleen in maintaining suppression of delayed-type hypersensitivity (DTH) after anterior chamber (AC) inoculation of allogeneic splenocytes. METHODS: Suppression of DTH response was tested in BALB/c mice after AC inoculation of allogeneic B10.D2 splenocytes. Seven days after AC injection, the antigen-inoculated eyes were enucleated or the spleens were removed. After enculeation or splenectomy at different time intervals, DTH responses in groups of the BALB/c mice were examined. Spleen components obtained from BALB/c mice that had been primed by B10.D2 splenocytes in the AC 7 days earlier were transferred intravenously to groups of naive syngeneic acceptors. At various intervals after adoptive transfer, variations of DTH responses were tested. RESULTS: Inoculation of B10.D2 splenocytes to the AC of BALB/c mice induced antigen-specific suppression of DTH. Either enucleation of the antigen-inoculated eyes or splenectomy weakened the DTH-suppressive effect within 5 weeks and abolished it within 9 weeks, whereas the mice retaining both antigen-inoculated eyes and spleens maintained longstanding DTH suppression. Adoptive transfer of spleen components to syngeneic acceptors demonstrated DTH suppression for only 3 weeks. CONCLUSIONS: The antigen-inoculated eye and spleen are required for long-standing suppression of DTH after AC inoculation of allogeneic splenocytes.

Adoptive Transfer↗
PubMed1997

[Focal cortical dysplasia and epilepsy surgery].

We conducted corticectomy in twenty-five patients with intractable partial epilepsy due to focal cortical dysplasia (FCD). MRI could not detect FCDs in three patients, interictal SPECT, however, revealed hypoperfusion corresponding to FCDs in two of these patients, while the FCD in one remaining patient was histologically identified in a resected specimen. The location of FCDs was as follows: the frontal lobe in sixteen patients, the temporal in five, the occipital in two, fronto-parietal in one, and the temporo-parietal in one. Prior to the surgery, twenty-one patients underwent invasive long-term intracranial EEG/CCTV monitoring. Of the fourteen patients who were tracked for longer than 2 years following surgery, eleven belonged to Class I according to Engel's criteria, two to Class II, and one to Class III. We compared intracranial EEG findings (interictal and ictal) between these twenty-one patients and eight patients with frontal lobe epilepsy resulting from different lesions. The results of this comparison, together with the seizure outcome following surgery, indicated that FCD is intrinsically epileptogenic. The invasive long-term monitoring should, as a rule, be performed in all patients with FCDs prior to the corticectomy.

Adolescent↗
PubMed1997

[Chronic myelomonocytic leukemia developed 7 years after the onset of idiopathic thrombocytopenic purpura like syndrome].

A 55-year old woman admitted to our hospital with bleeding tendency. She was diagnosed as having idiopathic thrombocytopenic purpura (ITP) by the platelet count 4.8 x 10(4)/microliter, Platelet associated IgG (PAIgG) 88.5 ng/10(7) cells, and an increase of megakaryocyte (81/microliter) of the sternal bone marrow. No obvious dysplasia of three lineages was observed. Because she did not respond to corticosteroid and gamma globulin, she was undertaken splenectomy 3 years after the diagnosis and the platelet count had been kept more than 3.0 x 10(4)/microliter during the following 2 years. After 7 years from the onset of ITP, she was admitted because of leukocytosis (16500 microliters with 8% monocytes) and thrombocytopenia (1.9 x 10(4)/microliter) with bleeding tendency. Hypercellular bone marrow with dysplasia of three lineages such as dyserythropoiesis, Pelger like nucleus, and micromegakaryocyte was observed. The chromosomal analysis presented 46XX, del (20) (q11.2) in all (50/50) cells. She was diagnosed as having chronic myelomonocytic leukemia (CMMoL). This is a difficult case in which it was distinguish ITP from refractory thrombocytopenia, a subtype of myelodysplastic syndrome. We reexamined and found some morphological abnormalities at diagnosis, suggesting that it might be preleukemic stage.

Blood Cell Count↗
PubMed1997

Malignant fibrous histiocytoma of the temporal bone with endocranial extension.

Cranial malignant fibrous histiocytomas are rare tumors. Most are hypervascular, destructive masses that are similar to other malignant lesions and to malignant fibrous histiocytomas found elsewhere in the body. We describe a myxoid malignant fibrous histiocytoma of the temporal bone, possibly of dural origin, with features that more closely resembled a meningioma at CT, MR imaging, and angiography.

Adult↗