Biomedical subjects

A Inoue

Publications and source records attributed to A Inoue.

At least 163 records · Page 9Linked to original sources

PubMed1997

Suppression of TNF-alpha secretion by azelastine in a rat mast (RBL-2H3) cell line: evidence for differential regulation of TNF-alpha release, transcription, and degranulation.

The mast cell plays a pivotal role in initiating allergic inflammation by secreting several cytokines including TNF-alpha, in addition to granule mediators such as histamine. Anti-allergic drugs including azelastine prevent immediate-type hypersensitivity by inhibiting mast cell degranulation, as well as blocking histamine H1 receptors. However, their effects on cytokine release from mast cells remain unknown. In a rat mast RBL-2H3 cell line, azelastine inhibited Ag- and ionomycin-induced TNF-alpha release with IC50 values of 25.7 +/- 3.4 microM and 1.66 +/- 0.45 microM, respectively. These effects were observed at lower concentrations than needed for the inhibition of degranulation. In Ag-stimulated cells, azelastine also inhibited TNF-alpha mRNA expression, TNF-alpha protein synthesis and release, and, possibly related to these effects, Ca2+ influx. In ionomycin-stimulated cells, however, azelastine inhibited TNF-alpha release to a greater extent than mRNA expression/protein synthesis and Ca2+ influx, suggesting that azelastine inhibits the release process more potently than transcription or production of TNF-alpha by interfering with a signal other than Ca2+. Azelastine added 1 h after ionomycin stimulation also immediately blocked subsequent release of TNF-alpha, which had been produced in the cells, without affecting Ca2+ influx. Pretreatment with 1 microM azelastine inhibited ionomycin-induced, but not Ag-induced, protein kinase C translocation to the membranes. These results suggest that the release process of TNF-alpha in mast cells is regulated by a mechanism distinct from that of degranulation, and that in Ca2+-ionophore-stimulated cells, it is also different from that of transcription/production, and possibly involves protein kinase C activation.

Animals↗

PubMed1997

Expression and mutational analysis of the DCC, DPC4, and MADR2/JV18-1 genes in neuroblastoma.

Loss of heterozygosity (LOH) on chromosome 18q21 is found frequently in various human cancers. Three candidate tumor suppressor genes, DCC (deleted in colorectal carcinomas), DPC4 (deleted in pancreatic carcinomas, locus 4), and MADR2/JV18-1 (MAD-related gene 2), have been cloned and identified from this chromosome region. We have reported recently that LOH on chromosome 18q is observed frequently in neuroblastoma. Alterations of DCC are involved in many human tumors. DPC4 and MADR2/JV18-1 are recently demonstrated to be altered in pancreatic and colorectal cancers, respectively. To confirm if inactivation of the DCC, DPC4, and MADR2/JV18-1 genes is involved in the pathogenesis of neuroblastoma and to clarify the mechanism of inactivation, we analyzed the expression of DCC, DPC4, and MADR2/JV18-1 in neuroblastoma cell lines and primary tumors by reverse transcription-PCR and investigated the mutations in the coding regions of these genes by PCR/reverse transcription-PCR single-strand conformation polymorphism. We found that 12 of 25 (48%) cell lines and 14 of 32 (44%) primary tumors, including 3 with 18q LOH, had absent or reduced expression of DCC mRNA. Expression was more likely to be reduced in advanced (67%) than in early stage neuroblastomas (24%) (P = 0.036), suggesting that inactivation of the DCC gene plays an important role in the progression of neuroblastoma. Altered expression of DPC4 was found in six (24%) cell lines and six (19%) tumors. MADR2/JV18-1 expression was reduced or absent only in four (16%) cell lines and three (9%) tumors. Mutations of the DCC genes were examined in 25 of 29 exons in neuroblastoma cell lines, and those exons in which mutations were found were further examined in primary tumors. We found missense mutations of AAC (Asn) to AGC (Ser) at DCC codon 176 in one cell line and ACC (Thr) to ATC (Ile) at codon 1105 in one cell line and tumor, respectively; polymorphisms of CGA (Arg) to GGA (Gly) at codon 201 and TTT (Phe) to TTG (Leu) at codon 951 in most of the cell lines and tumors; and a silent mutation of GAG (Glu) to GAA (Glu) at codon 118 in four cell lines and five primary tumors. We did not identify any mutations in the DPC4 and MADR2/JV18-1 genes in neuroblastoma. Our results suggested that mutations of the DCC gene may be involved in the pathogenesis of neuroblastomas but failed to account for the relatively high frequency of the altered expression, implying that other mechanisms are responsible for the inactivation of the DCC gene in neuroblastoma. Low frequency of reduced or absent mRNA expression and lack of mutations in DPC4 and MADR2/JV18-1 genes suggested a limited role for these two genes in neuroblastoma.

Cell Adhesion Molecules↗

PubMed1997

Microbial flora in the deepest sea mud of the Mariana Trench.

In an attempt to characterize the microbial flora on the deepest sea floor, we isolated thousands of microbes from a mud sample collected from the Mariana Trench. The microbial flora found at a depth of 10897 m was composed of actinomycetes, fungi, non-extremophilic bacteria, and various extremophilic bacteria such as alkaliphiles, thermophiles, and psychrophiles. Phylogenetic analysis of Mariana isolates based on 16S rDNA sequences revealed that a wide range of taxa were represented.

Bacteria↗

PubMed1997

Cloning and characterization of APS, an adaptor molecule containing PH and SH2 domains that is tyrosine phosphorylated upon B-cell receptor stimulation.

Stimulation of B lymphocytes through their antigen receptor (BCR) results in rapid increases in tyrosine phosphorylation of a number of proteins, which leads to a cascade of biochemical changes that initiates B cell proliferation and differentiation or growth inhibition. A novel cDNA, designed APS, encoding an adaptor protein with a Pleckstrin homology (PH) domain, Src homology 2 (SH2) domain, and a tyrosine phosphorylation site was cloned from a B cell cDNA library using a yeast two hybrid system. APS is structurally similar to SH2-B, an SH2 protein that potentially binds to the immunoreceptor tyrosine-based activation motif (ITAM) as well as Lnk which is postulated to be a signal transducer that links T-cell receptor to phospholipase Cgamma, Grb2 and phosphatidylinositol 3-kinase. APS expressed only in human Burkitt's lymphoma cells among cell lines we examined and tyrosine phosphorylated in response to BCR stimulation. APS bound to Shc irrespective of stimulation and bound to Grb2 after stimulation, suggesting that it plays a role in linkage from BCR to Shc/Grb2 pathway. These results indicate that APS, SH2-B and Lnk form a new adaptor family that links immune receptors to signaling pathways involved in tyrosine-phosphorylation.

Adaptor Proteins, Signal Transducing↗

PubMed1997

Lamina-specific connectivity in the brain: regulation by N-cadherin, neurotrophins, and glycoconjugates.

In the vertebrate brain, neurons grouped in parallel laminae receive distinct sets of synaptic inputs. In the avian optic tectum, arbors and synapses of most retinal axons are confined to 3 of 15 laminae. The adhesion molecule N-cadherin and cell surface glycoconjugates recognized by a plant lectin are selectively associated with these "retinorecipient" laminae. The lectin and a monoclonal antibody to N-cadherin perturbed laminar selectivity in distinct fashions. In contrast, neurotrophins increased the complexity of retinal arbors without affecting their laminar distribution. Thus, cell surface molecules and soluble trophic factors may collaborate to shape lamina-specific arbors in the brain, with the former predominantly affecting their position and the latter their size.

Animals↗

PubMed1997

Aripiprazole, a novel antipsychotic drug, inhibits quinpirole-evoked GTPase activity but does not up-regulate dopamine D2 receptor following repeated treatment in the rat striatum.

Aripiprazole, a quinolinone derivative, is a new dopaminergic agent which has been recently developed and demonstrated to be clinically useful as an antipsychotic drug with reduced extrapyramidal motor side effects. Here, we found that aripiprazole competed [3H]spiperone binding with a 100-fold higher affinity than [3H]SCH23390 binding, and inhibited the quinpirole-induced facilitation of high-affinity GTPase activity in rat striatal membranes. The effects of chronic administration of aripiprazole and haloperidol on dopamine D2 receptor binding and mRNA level in rat striata were examined by a [3H]spiperone binding assay and a ribonuclease protection assay. Haloperidol induced a significant rise in Bmax of [3H]spiperone binding at 1 mg/kg and in the level of dopamine D2L receptor mRNA at 4 mg/kg. A high dose of aripiprazole (100 mg/kg) only tended to increase the Bmax of [3H]spiperone binding non-significantly, and had no effect on the level of dopamine D2L receptor mRNA. These results indicated that aripiprazole had an antagonistic activity to dopamine D2 receptors with a high affinity, but that the potency of aripiprazole to up-regulate dopamine D2 receptors in the striatum was much smaller than that of haloperidol. This small up-regulation may be related to the ability to aripiprazole to act without side effects including tardive dyskinesia.

Animals↗

PubMed1997

Angiotensin II stimulates the proliferation of osteoblast-rich populations of cells from rat calvariae.

We examined the effects of angiotensin II (Ang II) on the proliferation of osteoblast-rich populations of cells obtained from calvariae of newborn rat. Addition of Ang II to the culture medium caused dose-dependent increases in the rate of DNA synthesis. Such increases were completely inhibited by the addition of DuP753, an antagonist of AT1 receptor. Ang II potentiated the production of inositol 1,4,5-trisphosphate (IP3) in the culture. Ang II also stimulated the activities of mitogen-activated protein kinases (MAPKs) upon binding to the AT1 receptor. These results suggest that Ang II might be intimately involved in the proliferation of the cells in calvariae through the AT1 receptor.

Angiotensin II↗

PubMed1997

Significance of granulation tissue in torn supraspinatus insertions: an immunohistochemical study with antibodies against interleukin-1 beta, cathepsin D, and matrix metalloprotease-1.

The pathophysiology of rotator cuff tears can be elucidated by examining the tendinous insertion of the supraspinatus muscle. As seen by light microscopy, the granulation tissue around the insertion of a torn supraspinatus tendon appears to induce osteochondral destruction by means of multinucleated giant cells and chemical mediators. The purpose of this study was to examine the contribution of certain chemical mediators to osteochondral destruction using immunohistochemical analysis of interleukin-beta, cathepsin D, and matrix metalloprotease-1. Sixteen supraspinatus insertions with portions of the greater tuberosity, including eight complete-thickness tears and eight incomplete-thickness tears, were obtained during surgery. Six fresh cadaveric supraspinatus tendons without grossly evident tears served as normal controls. Strong immunoreactivity was found in all 16 torn supraspinatus insertions but not in the six insertions of apparently intact tendons. Macrophages and multinucleated giant cells, which showed immunoreactivity for all three chemical mediators, were often found at the interface between the osteochondral margin of the enthesis and the granulation tissue, suggesting that they may be involved in osteochondral destruction. We therefore concluded that, in addition to repetitive subacromial impingement, this granulation tissue may contribute to the development of rotator cuff tears by weakening the insertion.

Adult↗

PubMed1997

Expression of MAGE genes in osteosarcoma.

Expression of MAGE genes that encode tumor-rejection antigens recognized by cytotoxic T lymphocytes with major histocompatibility complex class-I antigens was investigated in human osteosarcomas (20 cell lines and eight fresh tumor tissues). MAGE-1, 2, 3, 4, and 6 genes were expressed at the mRNA level in 11 (52.4%), 10 (47.6%), 10 (47.6%) one (4.8%), and 10 (47.6%) of 21 tumor cell lines, respectively, and in five (62.5%), six (75%), five (62.5%), one (12.5%), and five (62.5%) of eight fresh tumor tissues as determined by the reverse transcription-polymerase chain reaction method. MAGE-1 or 4 protein was detected by immunoblot analysis in eight of 11 or one of one tumor cell lines, respectively, where it was expressed at the mRNA level. Major histocompatibility complex class-I antigens were expressed in 19 of 21 tumor cell lines. These results suggest that MAGE tumor-rejection antigens are expressed in substantial numbers of osteosarcomas in a major histocompatibility class-I-restricted manner.

Antigens, Neoplasm↗

PubMed1997

Functional evaluation of hip abductor muscles with use of magnetic resonance imaging.

The hip abductor muscles are considered important for gait and biomechanics of the hip joint; however, their specific function has not been defined precisely. The intensity of magnetic resonance imaging signals in skeletal muscle has been reported to increase immediately after exercise. Making use of this phenomenon, we evaluated the hip abductor muscles. Magnetic resonance imaging was performed after isometric exercise of the hip abductor in three positions (20 degrees of abduction, neutral, and 20 degrees of adduction). The abduction force of the hip was measured with a dynamometer, and electromyographic measurements were made simultaneously for the same hip positions. Additionally, magnetic resonance imaging was performed after one-legged stance. As the hip was more adducted, the signal intensity increased on the scans. The values for muscle force, as evaluated with the dynamometer and integrated electromyography, also supported the results. The increase in signal intensity of the gluteus minimus at 20 degrees of abduction and after one-legged stance was significantly greater than that of the gluteus medius (p < 0.0001 and p < 0.0001, respectively). The results of this study indicate that the gluteus minimus muscle, along with the gluteus medius, plays an important role in hip abduction, gait, and stabilization of the pelvis.

Adult↗

PubMed1997

Metal-on-metal CoCrMo McKee-Farrar total hip arthroplasty: characteristics from a long-term follow-up study.

A total of 38 cemented metal-on-metal CoCrMo McKee-Farrar total hip arthroplasties (THAs) were clinically and radiographically evaluated over a long-term follow-up. No osteolysis and no granuloma were found more than 20 years after the operation. The main radiological findings were bone erosion and migration of the acetabular component, seen in 17 hips (44.7%). The direction of the migration correlated with the setting position of the acetabular component at operation. At revision surgery, metallosis was observed in unstable THA, and no metallosis was observed in stable THA. Using a micrometer, no wear of the sockets was found. Therefore, the loosening was thought to be due to the equatorial bearing rather than to metallosis. The mean survival of the THA to data was 14.6 years in those patients 60 years old or younger at operation and was significantly less (11.9 years) in those 61 years old or older at operation (P < 0.03). The mean survival time was 13.8 years.

Adult↗

PubMed1997

Correlation between vitamin D receptor genotypes and bone mineral density in Japanese patients with osteoporosis.

In order to better understand the pathogenesis of osteoporosis, we investigated the correlation between the vitamin D receptor (VDR) genotypes defined by BsmI restriction enzyme, as well as other related factors, and the bone mineral density (BMD) at the lumbar spine in 90 Japanese patients with osteoporosis. The same study was performed in 36 patients with osteoarthrosis of the hip joint and 92 healthy volunteers. The majority of the VDR genotypes were bb, and a few of the population showed either the BB or Bb genotype in all three groups. There was no statistical difference in the frequencies of these VDR genotypes in the three groups. The mean age-matched value of BMD (Z scores) at the lumbar spine in patients with osteoporosis was significantly lower than that in patients with osteoarthrosis or healthy volunteers. The mean Z scores of the healthy volunteers with bb genotype were significantly higher than those with BB genotype, whereas those of the osteoporosis patients with BB genotype were significantly higher than those with Bb genotype. There was no significant difference in the mean Z scores between bb and Bb genotypes in patients with osteoporosis and healthy volunteers. No significant difference was seen in the mean Z scores in patients with osteoarthrosis regardless of genotype. On the other hand, body weight significantly correlated with BMD in patients with osteoporosis by simple- and multiple-regression analysis. These results indicate that the BMD at the lumbar spine in Japanese patients with osteoporosis is affected by body weight, and might be affected partially by the VDR genotypes defined by BsmI.

Adult↗

PubMed1997

Detection of the soluble form of the Fas molecule in patients with multiple sclerosis and human T-lymphotropic virus type I-associated myelopathy.

We evaluated the presence of soluble Fas molecule (sFas) in the cerebrospinal fluids (CSF) and the sera of patients with multiple sclerosis (MS) or human T-lymphotropic virus type I (HTLV-I) associated myelopathy (HAM) using an enzyme-linked immunosorbent assay (ELISA). Patients with multiple sclerosis in the active phase had higher sFas serum levels than control (p < 0.005). In addition, significantly increased serum levels of sFas were found in patients with HAM (p < 0.005). We found a significantly increased CSF levels of sFas in patients with HAM and patients with MS in the active stage (p < 0.005). These results suggest that serum sFas may be related to clinical activity in patients with MS and that Fas may play an important role in the pathogenesis of HAM.

Adult↗

PubMed1997

Phosphatidylserine suppresses Theiler's murine encephalomyelitis virus-induced demyelinating disease.

Intracerebral inoculation of susceptible strains of mice with Theiler's murine encephalomyelitis virus (TMEV) results in immune-mediated demyelinating disease. Various cytokines may play an important role in the pathogenesis of this disease. Tumor necrosis factor (TNF)-alpha contributes to the further development of perivascular cellular infiltration and demyelination in the central nervous system (CNS). Phosphatidylserine (PS), a major anionic phospholipid of mammalian cells, has been proposed to function as a regulator of immune and inflammatory responses, especially reducing TNF-alpha production and release in mice. We studied the effect of PS on TMEV-induced demyelinating disease (TMEV-IDD). We injected TMEV intracerebrally into susceptible SJL/J mice and induced TMEV-IDD. PS were injected intraperitonealy, and clinical course and various immunological indicators were closely studied. The results show that when PS were administered in the effector phase. TMEV-IDD was significantly (P < 0.01) suppressed both clinically and histologically. In an ELISPOT assay, the number of TNF-alpha producing spleen cells was low in PS treated mice compared with saline treated control mice. mRNA of TNF-alpha was not detected in spleen cells of mice PS treated in the effector phase. These data suggest that administration of PS suppresses TMEV-IDD by suppressing TNF-alpha production in the effector phase.

Animals↗

PubMed1997

Fibrin deposition in the central nervous system correlates with the degree of Theiler's murine encephalomyelitis virus-induced demyelinating disease.

We examined the role of coagulation-fibrinolysis system in Theiler's murine encephalomyelitis virus-induced demyelinating disease (TMEV-IDD). The degree of fibrin deposition around the vessels in the spinal cord was significantly higher in susceptible SJL/J mice on 30 days post intracerebral injection (i.c.) than resistant C57BL/6 mice on 30 days post i.c. or mock infected SJL/J mice. Treatment with batroxobin (30 BU/kg/day), which is a thrombin-like defibrinogenating enzyme, causing a profound degree of afibrinogenemia, suppressed clinical signs of TMEV-IDD. Plasma fibrinogen concentration was significantly decreased in batroxobin-treated mice. Histologically, though the degree of perivascular mononuclear cell infiltration in the spinal cord was not suppressed in batroxobin-treated mice compared to saline-treated control mice, fibrin deposition was markedly suppressed in batroxobin-treated mice. These findings suggest that batroxobin suppresses TMEV-IDD through its defibrination effect, and provide evidence that CNS-associated deposition of fibrin and ensuing fibrinolysis, together with increased permeability of the blood-brain barrier (BBB), are prerequisite events for clinical manifestations of TMEV-IDD.

Animals↗

PubMed1997

Circulating matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-1 as serum markers of fibrosis in patients with chronic hepatitis C. Relationship to interferon response.

BACKGROUND/AIMS/METHODS: The imbalance between matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) is considered to be an important determinant of extracellular matrix deposition and breakdown. We measured serum MMP-1, MMP-2, TIMP-1 and TIMP-2 levels using the respective one-step sandwich enzyme immunoassays in 98 patients with chronic hepatitis C treated with interferon beta to examine their clinical significance for assessment of liver histology and to determine whether they can be useful as predictors of the interferon response. RESULTS: Serum TIMP-1 levels showed a positive correlation with the degree of fibrosis (r(s)=0.30, p= 0.004). Serum MMP-2 levels revealed positive relationships with the degree of periportal necrosis (r(s)= 0.32, p=0.002), the degree of fibrosis (r(s)=0.26, p= 0.01) and total score of histological activity index (r(s)=0.24, p=0.02). Serum MMP-2 levels were significantly higher in patients with no response than in those with sustained and transient response (p<0.01 and p<0.05, respectively), while serum MMP-1 levels did not differ among the three groups. Compared with the levels in sustained responders, the total amounts of serum TIMP-1 were significantly lower in transient responders and non-responders (p<0.01 and p<0.001, respectively). As for serum TIMP-2 levels, a significant decrease was found in transient responders and non-responders (p<0.01). The ratios of serum MMP-2 to TIMP-1 levels were significantly higher in transient responders and non-responders than in sustained responders (p<0.001, respectively) even when HCV RNA levels were low in patients with HCV genome subtype 1b or when the HCV genome subtype was 2a or 2b. Sustained response was never found in type 1b patients with ratios of serum MMP-2 to TIMP-1 levels of over 6.0. In logistic multivariate regression analysis, the ratios of serum MMP-2 to TIMP-1 level (p=0.0001), HCV genome subtype (p=0.005) and serum TIMP-2 level (p=0.03) were the independent predictors for sustained response, while serum MMP-2 level (p=0.0006) was the only predictor for no response. CONCLUSIONS: Serum MMP-2 and TIMP-1 levels might be useful for estimating the degree of liver fibrosis. The ratio of serum MMP-2 to TIMP-1 levels may serve as a new predictor of interferon response in patients with chronic hepatitis C.

Alanine Transaminase↗

PubMed1997

Alterations of the tumour suppressor gene DCC in neuroblastoma.

The deleted in colorectal carcinoma (DCC) gene, a candidate tumour suppressor, might be inactivated in a number of human cancers. In order to evaluate the possible role of DCC alterations in the pathogenesis of neuroblastoma, we examined 25 neuroblastoma cell lines and 16 primary tumours, including 6 samples with loss of heterozygosity (LOH) at the DCC locus for DCC mRNA expression, by using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique. The level of DCC expression was significantly reduced or undetectable in 12 of 25 (48%) cell lines and 7 of 16 (44%) primary tumours, suggesting that inactivation of the DCC gene is involved in the development of neuroblastoma. Three of the 6 tumours with LOH at the DCC locus revealed reduced DCC mRNA expression, indicating that LOH at the DCC locus might have affected the levels of DCC mRNA. We also screened for mutations in 4 exons of the DCC gene in 12 cell lines by using PCR-single strand conformation polymorphism (PCR-SSCP) analysis. Point mutations were not found except a polymorphic change at codon 201. The mechanism for inactivation of the DCC gene will be further investigated.

Genes, DCC↗

PubMed1997

Objective evaluation of pain in various spinal diseases: neuropeptide immunoreactivity in the cerebrospinal fluid.

A quantitative analysis was performed of substance P-like immunoreactivity (SPLI) and of beta-endorphin-like immunoreactivity (beta-ENDLI), in the cerebrospinal fluid (CSF) in various diseases. The results reported to date have not been consistent. The purpose of this study was to investigate whether or not the concentration of SPLI or that of beta-ENDLI in CSF demonstrated any potential for assessing the degree of subjective pain in various spinal diseases. SPLI in CSF was measured by radioimmunoassay in 158 patients with a spinal disease; involving 57 patients with a lumbar disc herniation (LDH), 38 with lumbar canal stenosis (LCS), 46 with cervical myelopathy (CM) and 17 with cervical radiculopathy (CR), and also in 20 healthy controls. beta-ENDLI in CSF was measured in 25 of these same patients; involving 12 with LDH, seven with LCS and six with CM, and also five of the same controls. The concentration of serum SPLI was also measured in 50 of these 158. The severity of pain was self-evaluated by each patient using a linear visual analogue scale (VAS). Their Japanese Orthopaedic Association (JOA) score was also calculated objectively using the clinical findings. Correlations were investigated among the concentrations of SPLI and beta-ENDLI in the CSF and the VAS and JOA clinical assessments of these patients. The concentration of SPLI in CSF was significantly higher in various spinal diseases than in control (P < 0.05), and was correlated with the severity on the VAS and with the JOA score. However, beta-ENDLI was not correlated with either the VAS or the JOA score. We conclude that the measurement of the SPLI concentration in CSF has the potential for assessing objectively the severity of pain associated with various spinal diseases.

Activities of Daily Living↗