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Biomedical subjects

A Inoue

Publications and source records attributed to A Inoue.

At least 127 records · Page 7Linked to original sources
PubMed1999

APS, an adaptor protein containing Pleckstrin homology (PH) and Src homology-2 (SH2) domains inhibits the JAK-STAT pathway in collaboration with c-Cbl.

We cloned a novel adaptor protein, APS (adaptor molecule containing Pleckstrin homology (PH) and Src Homology-2 (SH2) domains), which was tyrosine phosphorylated in response to c-kit or B cell receptor stimulation. Here, we report that APS was tyrosine phosphorylated by Janus kinase-2 (JAK2) at its C-terminal tyrosine residue and interacted with c-Cbl. Forced expression of APS in an erythropoietin (EPO)-dependent hematopoietic cell line resulted in reduced activation of STAT5 but not cell proliferation in response to EPO. APS bound to the phosphorylated tyrosine residue, Y343 of the erythropoietin receptor cytoplasmic domain. Co-expression of APS and c-Cbl, but not expression of either alone inhibited EPO-dependent STAT5 activation in 293 cells. This required the C-terminal phosphorylation site, as well as PH and SH2 domains of APS. Therefore, one of the major functions of APS is in recruitment of c-Cbl into the receptor/JAK complex, thereby inhibiting JAK signaling activity.

Adaptor Proteins, Signal Transducing↗
PubMed1999

Spondylolysis at three sites in the same lumbar vertebra.

We report a rare case of spondylolysis that occurred at 3 sites in the fifth lumbar vertebra involving the bilateral pars interarticularis and the center of the right lamina. This is the first such case in the world literature.

Adolescent↗
PubMed1999

Competitive polymerase chain reaction for the determination of N-myc amplification in neuroblastoma: report of clinical cases.

If an unfavorable prognosis is suspected in neuroblastoma, decision on a treatment protocol should be based on the N-myc copy number (12). We already demonstrated that the newly developed competitive polymerase chain reaction (competitive PCR) is a promising method for the determination of the N-myc copy number (6), and have started to use this competitive PCR procedure in neuroblastoma patients, together with fine-needle biopsy in selected cases. Seven children were studied. In one infant of 5 months of age whose tumor was diagnosed before undergoing mass screening for neuroblastoma, the competitive PCR procedure was performed with a fine-needle biopsy, and after obtaining a negative report on N-myc amplification within 48 hours, a regular protocol of treatment could be started without delay. We report that competitive PCR is a rapid and accurate method for the determination of the N-myc copy number, requiring only a small amount of material, and anticipate that competitive PCR will become the procedure of choice for the determination of N-myc copy number in neuroblastoma.

Adrenal Gland Neoplasms↗
PubMed1999

Serum levels of apoptosis-related molecules in patients with multiple sclerosis and human T-lymphotropic virus Type I-associated myelopathy.

We evaluated the presence of soluble Fas (sFas), Fas ligand (sFasL), and Bcl-2 in the sera of patients with multiple sclerosis (MS) or human T-lymphotropic virus type I (HTLV-1)-associated myelopathy (HAM) using an enzyme-linked immunosorbent assay (ELISA). Patients with MS in the active phase had higher sFas and Bcl-2 levels than had controls (sFas, p < 0.005; Bcl-2, p < 0.05) or patients in the inactive phase (p < 0.05). In addition, significantly increased serum levels of sFas were found in patients with HAM (p < 0.005). Interestingly, levels of sFasL in sera from patients with HAM and MS in the active stage were higher than those from controls or from patients with MS in the inactive stage or from other inflammatory neurologic diseases (OIND), although this was not statistically significant. These results suggest that serum sFas, sFasL, and Bcl-2 may play an important role in the pathogenesis of MS and HAM.

Adult↗
PubMed1999

Correlation between induction of expression of biglycan and mineralization by C-type natriuretic peptide in osteoblastic cells.

We reported previously that C-type natriuretic peptide (CNP) promotes the differentiation and mineralization of osteoblastic cells [Am. J. Physiol. 270 (Cell Physiol. 39): C1311-C1318, 1996]. However, little information is available about the mechanism of action of CNP in differentiating osteoblastic cells. Using the technique known as differential display-polymerase chain reaction, we attempted to identify the mRNAs whose levels are regulated by CNP in mouse clonal preosteoblastic MC3T3-E1 cells. One species of mRNA whose level was increased by CNP was 99% homologous to the 3'-untranslated region of a mouse gene for biglycan (BGN), a small proteoglycan. BGN is known to be involved in bone formation by osteoblastic cells. Therefore, we investigated the relationship, during the formation of mineralized nodules, between CNP and BGN using calvarial osteoblast-like cells (ROB cells) from newborn rats, that are a good model for studies on bone formation in vitro. Northern blot analysis revealed that transcription of the mRNA for BGN was up-regulated by CNP in ROB cells on days 6 and 8, whereas no effect of CNP was observed on days 3 and 12. Brief treatment with 10(-7) M CNP on days 3 through 9 exclusively enhanced the deposition of calcium, a result that suggests that CNP might regulate the expression of mineralization-related genes and, probably, the gene for BGN during a specific time period.

Animals↗
PubMed1999

Ipriflavone down-regulates endothelin receptor levels during differentiation of rat calvarial osteoblast-like cells.

Ipriflavone (7-isopropoxy-3-phenyl-4H-1-benzopyran-4-one) is a synthetic flavonoid that has been shown to stimulate the activity of osteoblasts. We show here that ipriflavone also promotes the deposition of calcium and the formation of mineralized nodules by newborn rat calvarial osteoblast-like (ROB) cells as well as the activity of alkaline phosphatase. We reported previously that endothelin-1 inhibits the differentiation of ROB cells [Y. Hiruma et al. (1998) J. Cardiovasc. Pharmacol. 31, S521-S523]. Therefore, we examined the effects of ipriflavone on the expression of endothelin receptors in ROB cells by polymerase chain reaction-Southern blot analysis and in binding assays with 125I-labeled endothelin-1. Ipriflavone reduced levels of endothelin ETA receptors (to 48% of the control level) in ROB cells around day 7 in our standard cultures, while it had no apparent effect on the expression of the mRNA for the endothelin ETA receptor. By contrast, treatment with 10(-7) M endothelin-1 on days 6 through 9 alone suppressed mineralization by ROB cells. Ipriflavone also reduced the ability of endothelin-1 to inhibit mineralization by ROB cells. These results suggest that the acceleration of osteoblastic differentiation by ipriflavone might be due, at least in part, to a time-specific down-regulation of endothelin receptors.

Animals↗
PubMed1999

Thermaerobacter marianensis gen. nov., sp. nov., an aerobic extremely thermophilic marine bacterium from the 11,000 m deep Mariana Trench.

A novel extremely thermophilic bacterium was isolated from the world's deepest sea-floor, the Mariana Trench Challenger Deep at a depth of 10,897 m. Cells were Gram-reaction variable, non-spore-forming and non-motile rods without flagella. Growth was observed between 50 and 80 degrees C (optimum: 74-76 degrees C; 90 min doubling time), pH 5.4 and 9.5 (optimum: pH 7.0-7.5) and 0.5 and 5% sea salts (optimum: 2% sea salts). The isolate was a strictly aerobic heterotroph capable of utilizing as sole energy and carbon source: yeast extract, peptone, cellulose, starch, chitin, casein, Casamino acids, a variety of sugars, carboxylic acids and amino acids. The G + C content of the genomic DNA was 72.5 mol%. Phylogenetic analysis based on 16S rRNA sequences placed this aerobic, high-G + C-content bacterium among the members of the Gram-positive, low-G + C-content anaerobic thermophilic bacteria within the Bacillus-Clostridium subphylum. On the basis of the physiological and molecular properties of the new isolate, the name Thermaerobacter marianensis gen. nov., sp. nov. (type strain 7p75aT = JCM 10246T) is proposed.

Bacteria, Aerobic↗
PubMed1999

Inhibition of experimental metastasis of human fibrosarcoma cells by anti-recombinant 37-kDa laminin binding protein antibody.

The laminin binding protein of 37 kDa (37LBP) is regarded as a precursor protein of the high-affinity 67-kDa laminin receptor (67LR). Expression of 67LR/37LBP is well correlated with biological aggressiveness of cancer, particularly with invasive and metastatic potential. To investigate in detail the role of 37LBP in cancer cells, we synthesized recombinant 37LBP (r37LBP) as a fusion protein and generated an IgG-type polyclonal antibody P4G against r37LBP. Western blot analysis with P4G showed a single band of 67LR under both nonreducing and reducing conditions using cell extract of human fibrosarcoma cells HT1080. It was shown that P4G inhibited cell attachment to immobilized laminin in a dose-dependent manner. Further, the intravenous injection of HT1080 cells pretreated with P4G, compared with that of cells pretreated with normal rabbit serum, resulted in a reduced number of experimental metastases (3.3+/-5.1 vs. 58.0+/-38.0 nodules per mouse, respectively) (P<0.005). These results suggest that P4G inhibits the colonization and growth of HT1080 cells in the lungs of mice, and that the blocking of r37LBP with the specific antibody P4G may offer a potential strategy for preventing cancer metastasis.

Amino Acid Sequence↗
PubMed1999

Detection of serum anticerebellar antibodies in patients with Miller Fisher syndrome.

We performed Western blot analysis to detect anticerebellar antibodies in the serum of patients with Miller Fisher syndrome (MFS). We studied 7 MFS patients, 6 Guillain-Barré syndrome (GBS) patients and 10 normal healthy persons as controls. Six MFS patients (86%) had IgG antibodies against mouse cerebellar protein, whereas 3 GBS patients (50%) and 4 healthy controls (40%) had antibodies. The mean number of antibodies in the serum of patient with MFS was 2.43, which was significantly more than that of GBS patients (mean 0.67) and healthy control (mean 0.70). This finding suggests variability and complexity of target in nervous systems that suffer aberrant immunity in MFS, and may also reflect the variability and heterogeneity of the pathogenesis of this syndrome.

Adrenal Cortex Hormones↗
PubMed1999

[Functional characterization of a sigma receptor and its gene expression by haloperidol].

Sigma (sigma) receptors are expressed in the brain as well as endocrine and immune systems. Several antipsychotic drugs such as haloperidol and pentazocine can bind to the sigma receptor, which is believed to play important roles in the pathogenesis of schizophrenia by as yet unknown mechanisms. Two subtypes of sigma receptors (sigma 1 and sigma 2) have been identified, and the sigma 1 receptor was cloned. The chronic administration of haloperidol to guinea pigs produced a marked inhibition of the binding to sigma 1 receptor, but did not change sigma 2-receptor binding. Scatchard analysis demonstrated that the inhibition was due to a reduction in the number of binding sites without changes in the affinity. The treatment with haloperidol also did not affect sigma 1-receptor mRNA detected by the RNase protection assay. The treatment of rats with haloperidol inhibited sigma 1-receptor binding to a much lesser extent than that to guinea pigs. These finding suggest that haloperidol or its metabolite, reduced haloperidol, which is produced in greater quantity in humans and guinea pigs than in rats and mice, might influence protein translation or modification of sigma 1-receptor without changing the transcriptional activity. The mechanisms through which sigma receptors could be differently regulated in vivo by chronic treatment with haloperidol may contribute to the therapeutic efficacy of haloperidol.

Animals↗
PubMed1999

Sequence analysis of a 32-kb region including the major ribosomal protein gene clusters from alkaliphilic Bacillus sp. strain C-125.

Forty-one open reading frames (ORFs) were identified in a 32-kb DNA fragment of alkaliphilic Bacillus sp. C-125. A similarity search using the BSORF database found 37 ORFs with significant sequence similarity to B. subtilis RNA polymerase subunits, elongation factor G, elongation factor Tu, and ribosomal proteins. Each ORF product showed more than 70% identity to those of B. subtilis. Gene organization in the region of str, S10, spc, and the alpha cluster was highly conserved among three strains, C-125, B. subtilis, and B. stearothermophilus.

Bacillus↗
PubMed1999

Aspergillus osteomyelitis in a child who has p67-phox-deficient chronic granulomatous disease.

Here we describe Aspergillus osteomyelitis of the tibia in a 9-year-old boy who has an autosomal recessive form of chronic granulomatous disease (CGD). The patient showed a p67-phagocyte oxidase (phox) deficiency, which is rare type of CGD in Japan. The initial treatment which consisted of surgical debridement and antibiotic therapy with amphotericin B (AMPH), did not control the infection. Aspergillus fumigatus (A. fumigatus) pure isolated from drainage fluid and necrotic bone tissue demonstrated less susceptible to antifungal agents, including AMPH, fluconazole and flucytosine. Recombinant interferon gamma was then administrated, and it was effective in controlling the course of severe invasive aspergillosis. This report indicates the use of interferon gamma might be helpful in control for Aspergillus osteomyelitis of the tibia in a child with CGD demonstrated p67-phox deficiency refractory to conventional therapy with AMPH.

Amphotericin B↗
PubMed1999

Efficacy of aquatic exercises for patients with low-back pain.

We have studied 35 patients (25 female and 10 male) with low-back pain who were managed with aquatic exercises after an appropriate period of treatment for their condition in the medical institution. The exercises employed consisted of strengthening exercises for the abdominal, gluteal, and leg muscles, stretching of the back, hip, hamstrings, and calf muscles, walking in water, and swimming. All the patients had been participating in the exercise program for more than 6 months. The frequency of performing exercises was once a week for 7 patients, twice a week for 19, and 3 or more times a week for the remaining patients. The method used in this study was a survey questionnaire which was composed of questions about the patient's physical and psychological condition. Those patients who had performed exercises twice or more in a week showed a more significant improvement in the physical score than those who performed exercises only once a week. More than 90% of the patients felt they had improved after 6 months of participation in the program. The improvement in physical score was independent of the initial ability in swimming. The results obtained suggested that exercises in water may be one of the most useful modes of exercise for a patient with low-back pain.

Adult↗
PubMed1999

Modulation of fas-mediated apoptosis in osteosarcoma cell lines.

Fas/APO-1 (or CD95) is a 45-kDa cell surface glycoprotein that transduces cellular death signals for apoptosis upon cross-linking with the Fas ligand, which is experimentally replaced by anti-Fas antibodies. We examined the Fas expression at the protein and mRNA levels, and susceptibility to anti-Fas-mediated apoptosis, in 14 osteosarcoma cell lines. Ten of 14 ostesarcoma cell lines constitutively expressed detectable levels of Fas on the cell surface with positive cell rates ranging from 10.2 to 72.4%. Four other cell lines expressed Fas almost exclusively in the cytoplasm. An antibody against Fas was able to induce Fas-mediated cell death only in two cell lines with high levels of surface Fas. In the presence of the protein synthesis inhibitor cyclohexamide, antibody to Fas led to an induction of apoptosis in ten of the osteosarcoma cell lines antibody-concentration dependently. These data suggest that Fas is a potential protein related to apoptosis in osteosarcomas when the sensitizing activity of cyclohexamide on anti-Fas-mediated cytotoxicity was exerted.

Antibodies, Monoclonal↗
PubMed1999

[A method for improving the accuracy in measurement of 99mTc-GSA liver uptake].

In order to improve the accuracy in the measurement of liver uptake rate of radioactivity at 15 minutes after injection of 99mTc-GSA, the corrected liver uptake rate (LU15VW) for tissue attenuation of gamma ray was measured. On the basis of the results of phantom studies, LU15VW was obtained as a ratio of the liver counts to the calculated counts of the injected dose supposed to homogeneously distribute in the liver of each patient and to decrease in count rate by the tissue attenuation of gamma ray, which was caused by the liver itself and body wall. The values of LU15VW were compared with those of the other 99mTc-GSA indices (LU15, LHL15, and HH15) and of the laboratory tests of liver function in 5 patients with chronic persistent hepatitis (CPH), 25 patients with chronic active hepatitis (CAH) 2A, 8 with CAH 2B, 8 with liver cirrhosis (LC), and 20 with hepatocellular carcinoma. LU15VW showed a good correlation with LU15, LHL15, and HH15 (r = 0.912, 0.864, and -0.869). The relationships between the results of LU15VW and the laboratory tests of liver function such as ICGR15, serum albumin, platelets counts, and hepaplastin test (r = -0.800, 0.684, 0.599, and 0.465) were more excellent as compared with those between the results of the other 99mTc-GSA indices and the laboratory tests. LU15VW was distinctly different in the mean values among the three groups of patients with CAH 2A, CAH 2B, and LC. These results indicated that LU15VW was an useful method for improvement of the accuracy in the measurement of liver uptake rate of 99mTc-GSA.

Humans↗
PubMed1999

[An animal model of corneal bacterial infection induced by intra-corneal injection of Staphylococcus epidermidis].

PURPOSE: Pharmacokinetic studies of anti-bacterial agents for infectious eye diseases have usually been performed using non-infected rabbit eyes with few studies using infected animal eyes. METHODS: In order to induce bacterial corneal inflammation in rabbit eyes, an intra-corneal injection of Staphylococcus epidermidis was done. The applied bacterial dose was 50 microliters of 1 x 10(4) cfu/ml. RESULTS: The characteristic corneal change was well-defined ulcer formation located in a limited area and relatively elevated. The area surrounding the infected part of the cornea was rather transparent. Using these infected eyes as models, the intraocular drug dynamics of 0.3% Ofloxacin were examined. The model was successfully induced in 78.6% of the rabbits. The drug concentration in the aqueous humor was 2 times higher in the infectious eyes than in the control eyes. CONCLUSION: The model might be useful for drug dynamic studies in infected eyes.

Animals↗
PubMed1999

Interleukin-1beta induces substance P release from primary afferent neurons through the cyclooxygenase-2 system.

Substance P (SP) is synthesized in the dorsal root ganglion (DRG) and released from primary afferent neurons to convey information regarding noxious stimuli. The effects of the proinflammatory cytokine interleukin-1 (IL-1) beta on the release of SP were investigated using primary cultured rat DRG cells. Recombinant mouse IL-1beta added to the cells at 0.1 ng/ml increased the SP-like immunoreactivity (SPLI) in the culture medium after incubation for 6 h by approximately 50% as compared with that of nontreated DRG cells. The effect of IL-1beta was Ca(2+)-dependent and significantly inhibited by 100 ng/ml IL-1 receptor-specific antagonist (IL-1r antagonist), cyclooxygenase (COX) inhibitors such as 0.1 mM aspirin, 1 microg/ml indomethacin, and 1 microM NS-398 (specific for COX-2), and 1 microM dexamethasone. Furthermore, a 1-h incubation with IL-1beta markedly increased the inducible COX-2 mRNA level, which was inhibited by an IL-1r antagonist and dexamethasone, whereas IL-1beta showed no effect on the level of constitutive COX-1 mRNA. These observations indicated that IL-1beta induced the release of SP from the DRG cells via specific IL-1 receptors, the mechanism of which might involve prostanoid systems produced by COX-2. This could be responsible for the hyperalgesic action with reference to inflammatory pain in the primary afferent neuron to spinal cord pathway.

Animals↗