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Biomedical subjects

A Inoue

Publications and source records attributed to A Inoue.

At least 235 records · Page 13Linked to original sources
PubMed1996

Formation of varanic acid, 3 alpha, 7 alpha, 12 alpha, 24-tetrahydroxy-5 beta-cholestanoic acid from 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholestanoic acid in Bombina orientalis.

Varanic acid (3 alpha, 7 alpha, 12 alpha, 24-tetrahydroxy-5 beta-cholestanoic acid; 24-OH-THCA) is almost the sole component of bile acids in the bile of Bombina orientalis. To examine in the mechanism of the formation of 24-OH-THCA, radiolabeled (25R)- and (25S)-3 alpha, 7 alpha, 12 alpha-trihdroxy-5 beta-cholestanoic acids [(25R)- and (25S)-THCA] and (24E)-3 alpha, 7 alpha, 12 alpha-trihdroxy-5 beta-cholest-24-enoic acid (delta 24-THCA) were administered intraperitoneally to B. orientalis, gallbladder bile was collected after 24 h, and bile acids were subsequently extracted. Then the bile acids were analyzed by means of radio thin-layer chromatography and radio high-performance liquid chromatography after conversion to p-bromophenacyl ester derivatives. Although delta 24-THCA was not converted to 24-OH-THCA, (25R)-THCA and (25S)-THCA were transformed to (24R,25R)-24-OH-THCA and (24R,25S)-24-OH-THCA, respectively. These results strongly suggest that 24-OH-THCA was transformed via direct hydroxylation of the saturated side chain of THCA, not via hydration to an alpha, beta-unsaturated acid, delta 24-THCA, in B. orientalis.

Animals↗
PubMed1996

Suppression of cell-transferred experimental autoimmune encephalomyelitis in defibrinated Lewis rats.

The role of coagulation-fibrinolysis system in experimental autoimmune encephalomyelitis (EAE) was studied by using batroxobin, derived from the venom of the South American pit viper Bothrops atrox moojeni. Batroxobin converts circulating fibrinogen into an insoluble form and causes a profound degree of afibrinogenemia. Batroxobin treatment (30 BU/kg/day) suppressed clinical signs of cell transferred EAE; the mean cumulative clinical score for batroxobin treated rats was 3.97, while saline treated controls scored 6.9 (P < 0.01). Plasma fibrinogen concentration decreased significantly in batroxobin-treated rats. Histologically, the degree of perivascular mononuclear cell infiltration in the spinal cord was not suppressed in batroxobin-treated rats compared to saline-treated control rats, however, deposition of fibrin around the vessels in the spinal cord was markedly suppressed in batroxobin-treated rats. These findings suggest that batroxobin suppresses EAE by preventing fibrin deposition, and provide evidence that CNS-associated deposition of fibrin and ensuing fibrinolysis, together with increased permeability of blood brain barrier (BBB), are related prerequisites for the clinical manifestation of EAE.

Animals↗
PubMed1996

Cervical myelopathy in elderly patients: clinical results and MRI findings before and after decompression surgery.

We examined 173 patients with cervical myelopathy of various causes. Seventy-seven patients underwent anterior decompression and fusion at not more than two levels, while 96 underwent posterior decompression by an expansive laminoplasty. Patients were followed up for between one and 4 1/2 years and the outcome was assessed both from a functional and a radiological point of view. The functional assessment used was according to the Japanese Orthopaedic Association (JOA) score (the higher the better), and the imaging outcome was assessed by a midline sagittal MRI assigned to three categories either for restoration of cord morphology, improvement or unchanged. Patients were divided into two groups: those 65 years old and older (50 patients), and those younger than 65 years old (123 patients). The data allowed the following conclusions to be reached: Older patients were likely to have more levels, and higher levels affected, and as a result were more likely to require a posterior operation. The recovery rate after an anterior operation was the same as that after a posterior operation. The pre- and post-operative JOA scores were higher in younger patients who tended to have milder disease with fewer levels affected. Younger patients had better morphological restoration. The recovery was likely to be better if the postoperative morphology was better. The morphology postoperatively was likely to be better if there had been less compression preoperatively. If the morphology was restored the disease duration was likely to have been less. The patients did better if the cord morphology was restored to normal, and this was easier to achieve in younger patients who had fewer levels involved and had less cord distortion preoperatively.

Adult↗
PubMed1996

Spinal deformity following surgery for spinal cord tumors and tumorous lesions: analysis based on an assessment of the spinal functional curve.

The mechanism of spinal deformity after surgical removal of a cervical spinal cord tumor or tumorous lesions was studied in 36 patients, based on the spinal functional curve prepared from the intersectional angle. The postoperative spinal deformity depends on the surgical level and the type of operation. In the laminectomy group, kyphosis of the upper cervical spine and compensatory increased lordosis of the lower cervical spine were observed in the C2 laminectomy patients. Localized kyphosis of the spine at the cervicothoracic junction and compensatory increased lordosis of the upper cervical vertebrae were noted in the C7 laminectomy patients. In the laminoplasty group, spinal deformities were less frequently observed, and when present the deformity was limited to a slight increase of lordosis, even in patients who had the facetectomy. These facts demonstrate the preventive effect of the laminoplasty regarding postoperative spinal deformity. Laminoplasty with reconstruction of the erector spinal muscles and the nuchal ligament is recommended for patients with a spinal cord tumor or a tumorous lesion. The spinal functional curve was significant in studying the biomechanics of the vertebral column with the advantage that both alignment and mobility of the spine are simultaneously, respectively and precisely visualized.

Adolescent↗
PubMed1996

Human astrocytoma cells (U-87 MG) exhibit a specific substance P binding site with the characteristics of an NK-1 receptor.

To investigate substance P (SP) receptors on an established human astrocytoma cell line (U-87 MG), [3H][Sar9,Met(O2)11]-SP, a selective SP receptor agonist, was used to identify and characterize the cell membrane binding sites for SP. SP receptor mRNA was examined by solution hybridization analysis, and the existence of SP binding protein on the surface of membranes was evaluated by flow cytometry using an anti-SP binding protein antibody. In U-87 MG and U-373 MG RNA preparations, transcripts were identified that corresponded to both mature and partially spliced receptor forms. In U-87 MG cell membrane-enriched preparations, the binding of [3H][Sar9,Met(O2)11]-SP was found to be time and cell number dependent, specific, saturable, and of high affinity. Equilibrium binding analysis revealed a single class of binding sites with an apparent KD of 1.15 +/- 0.15 nM and a Bmax of 108 +/- 9.8 fmol/mg of protein. [3H][Sar9, Met(O2)11]-SP binding was basically not influenced by addition of mono (Na+, Li+) or divalent (Mg2+, Mn2+, Ca2+) cations; only high doses of divalent cations decreased the binding. GTP and guanylyl-5'-imidodiphosphate, but not GDP and GMP, reduced the Bmax without changing the affinity of [3H][Sar9,Met(O2)11]-SP. We also examined the effects of pretreatment with three lectins [concanavalin A (con A), wheat germ agglutinin (WGA), and Lens culinaris agglutinin (LCA)] to determine the nature of carbohydrate chains on the U-87 MG cell. Of three lectins analyzed for effects on agonist binding, WGA and LCA had an inhibitory effect, whereas con A was ineffective. These results suggest that SP receptors on the human astrocytoma cell line U-87 MG have either a biantennary complex-type or a high mannose-type of carbohydrate chain and may be regulated by GTP-binding protein(s).

Astrocytoma↗
PubMed1996

Interleukin-4 stimulates rheumatoid synovial fibroblasts to express matrix metalloproteinase-1 (tissue collagenase) and histamine H1 receptor mRNA.

In the present study, we investigated the effect of interleukin-4 (IL-4) on metalloproteinase 1 (MMP-1/tissue collagenase) production in human rheumatoid synovial fibroblasts. Northern blot analysis revealed that addition of IL-4 with or without histamine stimulated the cells to increase the amount of proMMP-1 mRNA, and the IL-4 with histamine addition resulted in a 3.3-fold increase compared with histamine only. Furthermore, IL-4 itself stimulated the expression of histamine H1 receptor mRNA. These results suggest that IL-4 may play an important role in joint destruction in RA.

Arthritis, Rheumatoid↗
PubMed1996

The level of tumor necrosis factor-alpha producing cells in the spinal cord correlates with the degree of Theiler's murine encephalomyelitis virus-induced demyelinating disease.

The levels of tumor necrosis factor (TNF)-alpha producing cells were analyzed in mice with Theiler's murine encephalomyelitis virus-induced demyelinating disease (TMEV-IDD). Using an ELISPOT assay, we demonstrate an increase in TNF-alpha producing cells in the spinal cords of TMEV-infected SJL/J mice, especially at an active disease stage. The numbers of TNF-alpha producing cells were extremely high in susceptible SJL/J mice compared with the numbers in resistant BALB/c and C57BL/6 mice. TNF-alpha producing cells were also immunohistochemically identified in active lesions of TMEV-IDD at acute as well as chronic stages. The percentage of TNF-alpha producing cells compared with the total number of cells isolated from spinal cords was higher in TMEV-infected SJL/J mice than resistant BALB/c and C57BL/6 mice. Correspondingly, the level of TNF-alpha was much higher in the culture supernatants of both infiltrating cells in the spinal cords and spleen cells from clinically affected animals than that from similarly treated resistant mice. Treatment of virus-infected mice with a mAb specific for TNF-alpha at the beginning of the onset of disease suppressed the development of the demyelinating disease. These findings suggest that TNF-alpha may play an important role in the pathogenicity of TMEV-IDD.

Animals↗
PubMed1996

Change in the expression of C-type natriuretic peptide and its receptor, B-type natriuretic peptide receptor, during dedifferentiation of chondrocytes into fibroblast-like cells.

Chondrocytes derived from rat xiphoid cartilage dedifferentiated into fibroblast-like cells as the number of passages of the cells in culture increased. During in vitro dedifferentiation the growth of the cells was markedly suppressed. We had proposed previously that C-type natriuretic peptide (CNP) might be a potent antimitogenic factor for chondrocytes, and TGF-beta 1 induced a marked increase in CNP secretion of chondrocytes. Therefore, we investigated the expression of CNP, B-type natriuretic peptide receptor (NPR-B or GC-B), and TGF-beta 1 in this process. Radioimmunoassay and RNase protection analyses revealed passage-associated increase in CNP-like immunoreactivity and in levels of NPR-B mRNA, respectively. Northern blot analyses showed that the level of TGF-beta 1 mRNA decreased with increasing passage number. These results suggest that the expression of CNP and NPR-B might be involved in in vitro dedifferentiation of chondrocytes and TGF-beta 1 does not affect the increasing level of CNP during in vitro dedifferentiation.

Animals↗
PubMed1996

Structure of heads A and B of myosin studied by tryptic digestion of myosin subfragment-1.

We studied the difference in the structure of head B (P1-burst head) and head A of myosin by limited tryptic digestion of myosin subfragment-1 (S-1), and using antibodies (anti-A and anti-B) which bind specifically with each head. The antibodies were prepared using peptides with sequences identical to those around the reactive lysine residue of heads A and B. When myosin subfragment-1 (S-1) was cleaved limitedly by trypsin, S-1 heavy chain (100 kDa) was digested into fragments of 25, 50, and 20 kDa. Two fragments with molecular masses of 75 and 27 kDa were transiently produced in the initial phase of digestion. Anti-A and anti-B antibodies bound only with peptides that contained the reactive lysine residue [S-1 heavy chain (100 kDa), 75-, 27-, and 25-kDa peptides], thus showing specific binding with antigen peptide. However, the 27-kDa fragment bound more strongly with anti-B antibody than with anti-A antibody. When S-1 was separated into fractions rich in S-1A and S-1B using insoluble anti-A or anti-B antibody, each antibody bound more strongly with the S-1 heavy chain (100 kDa) of its corresponding fraction by Western immunoblotting. These results suggest that the antibodies react specifically with peptides even after SDS-PAGE and membrane-blotting, and that the structure of the 25 kDa 50 kDa junction differs between heads A and B of myosin.

Animals↗
PubMed1996

Cloning and sequencing of a cDNA for Akazara scallop troponin T.

A cDNA clone encoding troponin T of Akazara scallop (Chlamys nipponensis akazara) striated adductor muscle has been isolated and sequenced. The complete sequence deduced consists of 314 amino acid residues with a molecular weight of 37,206. Akazara scallop troponin T contains 55 amino acid residues more and 82 residues fewer than rabbit skeletal muscle troponin T and Drosophila melanogaster troponin T, respectively, showing almost the lowest sequence homology with rabbit troponin T (26%) but the highest homology with Drosophila troponin T (33%). Further, high sequence homology was seen in the functional regions: residues 33-120 and 174-227, corresponding respectively to residues 71-158 and 197-250 of rabbit troponin T (tropomyosin-binding regions); and residues 200-204, corresponding to 223 227 of rabbit troponin T (troponin I-binding region). In residues 1-70 (tropomyosin-binding region), however, only six residues are identical with rabbit troponin T.

Amino Acid Sequence↗
PubMed1996

Production of IL-6 by T cells from the femoral head of patients with rapidly destructive coxopathy (RDC).

RDC is a syndrome with unknown etiology that causes rapid destruction of a hip joint. We have investigated the production of osteoclast-activating cytokines (IL-6, IL-1alpha and tumour necrosis factor-alpha (TNF-alpha)), interferon-gamma (IFN-gamma) and IL-8 by T cells in the affected joint. The level of IL-6 produced by the T cell lines (TCL) established from the femoral head was significantly higher than that from patients' or healthy donors' peripheral blood mononuclear cells (PBMC). IL-6 production by the TCL from synovial membrane or from patients' PBMC was also significantly higher than that from healthy donors' PBMC. IL-1alpha production by the TCL from the femoral head was significantly higher than any of the other groups when all the TCL were used for the analysis. TNF-alpha production was highest in the TCL from patients' PBMC. The levels of IFN-gamma or IL-8 were not significantly different among these four groups. The plasma levels of all these cytokines except for IFN-gamma, that was rather lower, in RDC patients were not significantly different from those in osteoarthrosis or trauma patients, or healthy donors. These results suggest that T cells at the affected femoral head, and also synovial membrane to some extent, are involved in bone resorption through the production of IL-6 and probably IL-1alpha in patients with RDC.

Aged↗
PubMed1996

cGMP produced in response to ANP and CNP regulates proliferation and differentiation of osteoblastic cells.

The effects of natriuretic peptides on the proliferation and differentiation of osteoblast-like cells from rat calvariae were examined. Natriuretic peptides are physiological agonists that activate receptor guanylate cyclases, namely, natriuretic peptide receptor (NPR)-A and NPR-B. Exposure of cells to atrial natriuretic peptide (ANP) and C-type natriuretic peptide (CNP) resulted in large increases in the rate of intracellular production of guanosine 3',5'-cyclic monophosphate (cGMP). Moreover, CNP-like immunoreactivity was detected in the conditioned medium from osteoblast-like cells, while ANP was undetectable. In cells exposed to natriuretic peptides, a dose-dependent reduction in the rate of DNA synthesis was observed. Natriuretic peptides also stimulated the activity of alkaline phosphatase (ALPase) and the expression of mRNA for ALPase and osteocalcin and the mineralization of nodules by the cultured cells. These results could be reproduced by treating cells with 8-bromo-cGMP. Endothelin-1, whose physiological functions are the opposite of those of natriuretic peptides, decreased the ALPase activity and the mineralization of nodules. In the present study, natriuretic peptides were demonstrated to promote bone formation via the action of cGMP in a signal-transduction pathway mediated by specific receptors in osteoblast-like cells.

Alkaline Phosphatase↗
PubMed1996

Genetic clinical markers of human neuroblastoma with special reference to N-myc oncogene: amplified or not amplified?--An overview.

Neuroblastoma is the most common extracranial tumor in children, and cytogenetically, chromosome 1p deletions, extrachromosomal double minutes, and homogeneously staining regions (HSRs) are commonly observed in cell lines and in tumors in advanced stages. It is found that an HSR represents genomic amplification of N-myc, which plays a key role in determining the aggressiveness of neuroblastoma. However, stage IV neuroblastomas or cell lines which lack N-myc amplification are also progressive, and some of them show evidence of N-myc expression in terms of mRNA and/or N-Myc oncoprotein. It was recently shown that a small proximal locus mapped between 1p35-36.1 and 1p36.23 may function as a suppressor gene of N-myc amplification. In neuroblastoma, a pattern of diploidy is associated with rapid tumor growth and poor survival. Expression of bcl-2 proto-oncogene is strongly associated with unfavorable histology, while expressions of Ha-ras and trk-A proto-oncogenes indicate a favorable prognosis. trk-A proto-oncogene encodes a receptor for nerve growth factor. Genetic characteristics of neuroblastomas found by urinary catecholamine mass screening are also discussed.

Biomarkers, Tumor↗
PubMed1996

Competitive polymerase chain reaction for the quantification of N-myc gene copy number in neuroblastoma.

An absolute quantification method for the N-myc gene copy number of neuroblastoma specimens was established by applying the competitive polymerase chain reaction (cPCR). The competitor plasmid (pZH2) lacking an MluI site in the exon 2 was constructed to distinguish two product species amplified from genomic DNA and the competitor plasmid. By using this cPCR system, we could obtain qualitative results within 1 day, i.e. amplified or unamplified, and quantitative results by using radiolabelled nucleotides within 4 days. The copy numbers of N-myc in 47 neuroblastoma specimens by cPCR correlated well with those by Southern hybridization (r = 0.85). We conclude that cPCR is a simple and rapid method, requires only a small amount (200 ng) of sample DNA, and is expected to be used for prognostic evaluation in neuroblastomas.

Base Sequence↗
PubMed1996

S-methylmethionine sulfonium in fruits of citrus hybrids.

The S-methylmethionine sulfonium (MMS) concentrations in fruits of citrus hybrids were measured, and found to increase during ripening of the fruit. However, there of eleven hybrids of 'Seto unshiu' crossed with 'Morita ponkan' and four of 9 hybrids of 'Murcott' tangor crossed with 'Seto unshiu' had low MMS concentrations even at late harvest stage. Crossbreeding is useful in producing new citrus fruits that have juices with the desirable characteristics of their parents without formation of dimethyl sulfide which is an off-flavor.

Citrus↗
PubMed1996

Flushing pattern and idiopathic avascular necrosis of the femoral head.

A cooperative hospital-based case-control study of idiopathic avascular necrosis of the femoral head (IANF) was carried out to clarify the involvement of smoking, drinking, flushing pattern, and other factors in the development of IANF comparing 90 cases (64 males and 26 females) without history of systemic corticosteroid use with 180 matched controls (128 males and 52 females). The results of analyses were represented only for male subjects because of small number of female cases. There was no significant difference in smoking habits, daily and cumulative number of cigarettes smoked between case and control groups. Current drinkers had obviously higher risk (OR = 11.47) of IANF compared to nondrinkers or exdrinkers. In addition, there was a consistent risk increase with increasing alcohol consumption and the highly significant dose-response relationship remained unchanged after adjustment for all other factors (chi 2 = 14.33, p < 0.001 and chi 2 = 13.24, p < 0.001 for daily and cumulative alcohol consumption, respectively). For flushing pattern, although nonflushers had a significantly elevated risk (OR = 2.08) in the univariate analysis, the association disappeared (OR = 0.73) after adjustment for alcohol and other factors. Since nonflushers tend to be heavy drinkers, perhaps, an apparent risk increase among nonflushers may be due to alcohol drinking. Body mass index (BMI) was inversely related to the development of IANF. The risk reduction was found among subjects with higher BMI and an adjusted linear trend of OR was significant (chi 2 = 6.65, p < 0.05). However, further studies were required regarding the association between flushing pattern or BMI and IANF because of a few reports. History of liver diseases and occupational history were not significantly associated with the development of IANF after adjustment for other factors.

Adult↗
PubMed1996

[How to determine the best combination assay by several tumor markers?].

Combination assay by several tumor markers provides more precise diagnosis of malignancy than using a single tumor marker. The best combination assay by two tumor markers is determined as follows. MINIMIZE RFP (1-SpAorB(CA, CB)) + RFN(1-SnAorB(CA, CB)), where RFP, RFN denote the loss of false positive and false negative, respectively. SpAorB (CA, CB), SnAorB (CA, CB) are specificity and sensitivity of the combination assay by tumor markers A, B with cut-off values CA, CB.

Biomarkers, Tumor↗