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Biomedical subjects

A Takeda

Publications and source records attributed to A Takeda.

At least 325 records · Page 18Linked to original sources

NMDA receptor (NMDAR1) expression in the rat hippocampus after forebrain ischemia.

The influence of transient forebrain ischemia on the temporal alteration of NMDA receptor (NMDAR1) mRNAs in the hippocampal formation was analysed by in situ hybridization, RNA blot and in vitro receptor autoradiography ([3H]3-((+)2-carboxypiperazin-4-yl)-propyl-1- phosphonic acid). In the CA1 subfield of the hippocampus, decrease of NMDAR1 mRNA levels was not observed until death of the CA1 pyramidal cells. Reactive astrocytes, which appear after CA1 pyramidal cell death, expressed few NMDAR1. The CA3 and dentate gyrus also showed a reduction of NMDAR1 mRNA levels 7 days after ischemia. Temporal profiles of NMDAR1 mRNA levels correlated well with those of NMDA receptor-binding in the CA1 and CA3/dentate gyrus. These results indicate that NMDA receptors in the CA1 are predominantly located on the CA1 pyramidal cells and that these receptors are preserved during early recirculation period. Histologically intact neurons in the CA3 and dentate gyrus also had marked modulation in glutamatergic neurotransmission after ischemia.

Animals↗

Brain uptake of trace metals, zinc and manganese, in rats.

The brains of rats injected intravenously with 65ZnCl2 or 54MnCl2 were subjected to high resolution autoradiography. 65Zn and 54Mn were largely concentrated in choroid plexus 1 h after injection and then gradually decreased, with increases in other brain regions, suggesting that both metals were taken up gradually into brain mainly via cerebrospinal fluid in the choroid plexus. By 3 days after injection, relatively high level of 65Zn was seen in the dentate gyrus and CA3 region of the hippocampus and in the cerebral cortex. The level of 54Mn was also high in the former, while relatively low in the latter.

Animals↗

Molecular cloning of the CD45-associated 30-kDa protein.

CD45, a leukocyte-specific transmembrane protein tyrosine phosphatase, mediates signal transduction pathways critical for immune responses. However, the mechanism of CD45-mediated signal transduction and the identity of CD45-associated proteins have remained unclear. A CD45-associated 30-kDA phosphorylated protein (CD45-AP) was purified by virtue of its specific association with CD45, and its mouse cDNA was cloned by using the internal amino acid sequence information. In vitro translated CD45-AP bound specifically to CD45. CD45-AP appears to be leukocyte-specific and shares no significant homology with presently known sequences. The predicted sequence contains no consensus tyrosine phosphorylation sites or conserved sequences of GTP-binding proteins. CD45-AP may act as an adapter molecule for CD45-mediated signal transduction.

Amino Acid Sequence↗

Monoclonal antibodies as probes to detect conformational changes in the rat cysteine proteinase inhibitor cystatin A.

Five monoclonal antibodies (MAbs), 77, 114, 138, 175 and 187, were established for rat cystatin A. MAbs 77, 114, 138 and 175 were shown to belong to the IgG1 subclass, whereas MAb 187 was an IgM. These MAbs partially suppressed inhibitory activity of rat cystatin A to papain. Their epitopes were mapped in detail on the molecule by examining the reactivities of the MAbs with NH2-terminally truncated forms and fragments of rat cystatin A by an enzyme-linked immunosorbent assay (ELISA), and by reactivity with the inhibitor on immunoblotting. In competitive binding assays the MAbs did not compete with each other, indicating that the epitopes recognized by these MAbs were substantially different. The conformational epitope recognized by the three MAbs 114, 138 and 175 belonged to one group that was highly sensitive to denaturation, but those epitopes were unchanged by NH2-terminal truncation. MAb 187 was able to recognize a linear epitope present in amino acid residues 15-50 in the NH2-terminal region. MAbs 77 and 114 reacted weakly with mouse cystatin A but not at all with human cystatin A, whereas MAb 187 reacted similarly with mouse cystatin A but at about half that level with human. The MAbs produced in this study should be useful tools for detecting conformational changes in the rat cystatin A molecule.

Amino Acid Sequence↗

Intra-arterial chemotherapy using a reservoir for endocrine-refractory prostate cancer.

For local control in patients with endocrine-refractory prostate cancer, an intra-arterial chemotherapy regimen comprising methotrexate (MTX), Adriamycin (ADM), and cisplatin (CDDP) was evaluated. A total of 19 patients having a mean age of 66.4 +/- 8.8 years and a mean performance status (PS) of 1.3 +/- 1.0 were enrolled. Of these patients, 3 had proved to be resistant to initial endocrine therapy and the remaining 16 had relapsed from disease stabilization after endocrine therapy. The catheter tip was placed in the internal iliac artery in 16 cases, in the common iliac artery in 2 cases, and in the aorta in 1 case after occlusion of the contralateral feeding artery. The intraarterial chemotherapy was performed mainly using MTX (30 mg/m2), ADM (30 mg/m2), and CDDP (50 mg/m2) as one course and was repeated for a mean of 2.9 +/- 2.3 courses. Then, in an outpatient clinic, 5-fluorouracil (5-FU), ADM, or MTX was given intra-arterially as maintenance chemotherapy until re-relapse. As based on the criteria for evaluation of nonsurgical therapy in prostate cancer proposed by the Japanese Urological Association, the prostatic lesion showed a partial response (PR) in 9 cases and no change (NC) in 10 cases. As judged from the response of prostate-specific antigen (PSA), a complete response (CR) was obtained in 6 cases, a PR, in 3 cases; and NC and progressive disease (PD), in 2 cases each. Therefore, the overall response rate was 63%. Improvement in the symptoms was observed in 83% of patients. The duration of the response was 15.1 +/- 10.5 months for the PR cases and 7.4 +/- 5.7 months for the NC cases. Furthermore, the mean survival time observed in the PR group was 38.9 months, which was better than that seen in the NC (16.4 months) and PD (10.5 months) groups. These results suggest that intra-arterial chemotherapy may become an option for the treatment of locally advanced and endocrine-refractory prostate cancers. Using a reservoir, this chemotherapy can be easily given in an outpatient clinic.

Abdominal Muscles↗

Tumor detection by radioimaging of hepatic radiozinc--correlation with metallothionein content.

In order to determine if the radioimaging of the level of hepatic metallothionein (MT) is able to detect malignancy, we performed, using 45Zn, a trial to measure MT levels in the liver of experimental tumor-bearing mice and rats. The distribution of 65Zn in the livers of the mice and rats increased with s.c. transplantation of Ehrlich carcinoma and ascites hepatoma 7974F, respectively. A Sephadex G-75 elution profile of the cytosol fraction of the liver showed that the elevation of 65Zn level was due to elevation of the MT level. At 2 days after tumor transplantation in mice, the 65Zn-image demonstrated elevation of the hepatic MT level, while 67Ga-citrate, which is currently utilized, could not image the tumor at such an early period of growth. These results suggest that hepatic MT is a useful marker for the detection of malignancy at the early stage.

Animals↗

Antisense oligonucleotide of c-myc discriminates between zinc- and dexamethasone-induced synthesis of metallothionein.

The metallothionein II genes, whose structures are highly conserved throughout the animal kingdom, are composed of three exons and two introns. By using synthetic oligonucleotides with sequences complementary to the mRNA coding for human metallothionein II, we have shown that (a) inhibition of metallothionein synthesis causes cells to die from metal toxicity, (b) metallothionein possesses an essential gene, and (c) modifications in oligonucleotide structures exhibit specificity in inhibiting metallothionein synthesis. Furthermore, we have prepared a synthetic antisense oligodeoxyribonucleotide to the mRNA specific for human c-myc and tested its potential to regulate metallothionein synthesis in Chang liver cells in culture. The results of this study revealed that the c-myc antisense oligodeoxyribonucleotide led to greater induction of zinc-promoted but not of cadmium- or dexamethasone-induced synthesis of metallothionein. These data are interpreted to suggest that c-myc acts as a repressor of at least one of the six human metallothionein isoforms and demonstrates a unique mode of regulation capable of discriminating between zinc- and dexamethasone-induced synthesis of metallothionein.

Base Sequence↗

Immunoelectron microscopic localization of C-terminus of 43-kDa dystrophin-associated glycoprotein in normal human skeletal myofibers.

Ultrastructural localization of the C-terminus of a 43-kDa dystrophin-associated glycoprotein (43-DAG) and its relation to dystrophin were investigated in normal human skeletal myofibers by using single and double immunogold labelling methods. The C-terminal end of 43-DAG antibody was localized ultrastructurally at the inside surface of muscle plasma membrane and the sarcoplasmic side of the plasma membrane invaginations. Double immunolabelling electron microscopy disclosed that signals of the C-terminal end of 43-DAG and dystrophin were often associated with each other and formed a doublet at the inside surface of plasma membranes in normal human skeletal myofibers.

Cytoskeletal Proteins↗

Fabry-like laminated myelin body associated with IgA nephropathy.

We present the first female patient to exhibit Fabry-like myelin bodies in the glomerular epithelial cell in association with IgA nephropathy. This previously healthy 36-year-old woman presented with proteinuria and hematuria without skin lesions. Renal biopsy showed typical IgA nephropathy, with paramesangial deposits, mesangial proliferation and scattered myelin bodies. The leukocytic alpha-galactosidase A activity was abnormally low. She had no family history of Fabry's disease nor the characteristic features, such as skin lesion, neuralgia, or hypohidrosis. Fabry's disease is diagnosed from the renal biopsy findings and the activity of alpha-galactosidase A in leukocytes and/or fibroblasts. We diagnosed the present case with Fabry' disease and IgA nephropathy from these results.

Adult↗

Renal complications during pregnancy in a patient with diabetes mellitus.

We present a 23-year-old woman with a 7-year history of insulin-dependent diabetes mellitus (IDDM) who became pregnant. At the 23rd week of pregnancy she exhibited the signs and symptoms (hypertension, edema, proteinuria) of both diabetic nephropathy and preeclampsia. A cesarean section was successfully performed. The proteinuria persisted for more than 3 months after delivery. Renal biopsy confirmed the diagnosis of diabetic glomerulosclerosis together with the renal findings attributable to preeclampsia. The rapid acceleration of diabetic nephropathy in this patient was attributed to preeclampsia. We therefore recommend that patients with DM be followed closely during pregnancy in an attempt to prevent the acceleration of renal damage by preeclampsia.

Adult↗

[Nephropathy in patients with mixed connective tissue disease].

The first report of mixed connective tissue disease (MCTD) was described that nephropathy was rare complication of the patients with MCTD. Recently, some authors reported that the nephropathy was 25-50% of patients with MCTD. This report investigated the histopathological findings of patients with MCTD. This study was conducted on 17 patients associated with MCTD. In clinical features, there were six of nephrotic syndrome and two of hematuria and proteinuria. They were normal renal function, expect for one case (case MF). In pathological findings, there were four of membranous nephropathy, two of mesangial proliferative glomerulonephritis, including one of IgA nephropathy, and 11 of minor glomerular abnormality. Three cases translated to systemic lupus erythematosus (SLE) during follow-up period (1 to 5 years). They had hypocomplementemia and membranous nephropathy in common. In conclusion, the nephropathy is not rare complication in patients with MCTD. Sex of the 17 patients (35.3%) with MCTD have nephropathy. MCTD cases with immune-complex glomerulonephritis are likely to transit from MCTD to SLE.

Adolescent↗

[Clinical study of neoadjuvant intraarterial chemotherapy (I A-MAC) for advanced bladder cancer].

The clinical efficacy of intraarterial neoadjuvant chemotherapy with MTX, ADR and CDDP in 88 patients with advanced bladder cancer (> or = T2 or TCC G3) was evaluated. The intraarterial chemotherapy was performed using Seldinger's method. The three drugs were administered at the same time among 60 patients. In sixteen patients MTX was administered intravenously before intraarterial infusion of ADR and CDDP. In twelve patients, a high dose of CDDP (more than 100 mg/m2) was administered. The mean regression rate was 70%. That of the high-dose CDDP group was rather superior to the other two groups. However, the adverse reaction and duration of recovery were inferior to those of the other two groups. When intraarterial chemotherapy is performed as a neoadjuvant chemotherapy, care must be taken to determine the suitable dose.

Antineoplastic Combined Chemotherapy Protocols↗

[Quantity-frequency scale (QF Scale) for adolescent problem drinking].

From 1990 to 1992, we surveyed 14,438 Japanese high school students from nine prefectures on drinking behavior. Based on this survey, we developed a new Quantity-Frequency Scale (QF Scale) for adolescent problem drinking. Each adolescent is scored from 0 to 6 by the QF Scale, which is composed of scores of drinking quantities and frequencies. Adolescents were assessed as normal adolescent, drinkers and problem drinkers by QF Scale scores. The normal adolescent (score 0) categorized by the QF Scale drinks none or little and obeys the Law Prohibiting Adolescents from Drinking. The drinker (score 1-3) categorized by the QF Scale drinks at least several times a year, but drinks little each time. The problem drinker (score 4-6) categorized by the QF Scale drinks several times a week and has at least two drink (1 drink is equivalent to a glass of beer) each time, once a week with at least 3 drinks, or once or twice a month with at least 6 drinks each time. The problem drinkers show a physical, psychological and social high risk for alcohol. The validity of the QF Scale was examined. The results showed that the problem drinkers had significantly frequent blackout, significantly frequent drinking when they had emotional conflicts or they were alone, and drank significantly more hard liquor than the drinkers and normal adolescents. Among the high school students surveyed, 39.5% were normal adolescents, 43.2% were drinkers and 17.4% were problem drinkers. In conclusion, the QF Scale is a useful instrument for assessment of adolescent problem drinking.

Adolescent↗

[Nephropathy in patients with primary antiphospholipid antibody syndrome].

Primary antiphospholipid antibody syndrome (APS) is characterized by abortion, thrombosis, thrombocytopenia and/or valvular disease and it is liable to complicate systemic lupus erythematosus (SLE). We carried out a study to investigate the clinical and renal pathological findings in five patients with APS, but not SLE. In the clinical findings, the patients had negative tests for proteinuria and hematuria, and their renal function and tubular function were within normal limits. In the light microscopic findings, three patients exhibited mild mesangial hypercellularity, and two had minor glomerular abnormalities. In immunofluorescent study, there were IgM and/or C3 depositions in the mesangial area in some cases, and in electron microscopic study, there were no special findings other than mesangial hypercellularity. In conclusion, nephropathy is a rare complication in patients with APS, unlike systemic lupus erythematosus.

Adult↗

The bcl-2 protein is inserted into the outer membrane but not into the inner membrane of rat liver mitochondria in vitro.

The bcl-2 gene encodes a 26 kDa protein, overexpression of which blocks cell apoptosis. Since conflicting data have been reported concerning the localization of the Bcl-2 protein with mitochondrial inner and outer membranes, we analyzed in vitro import of the human Bcl-2 protein into isolated rat liver mitochondria. The Bcl-2 protein translated in rabbit reticulocyte lysate was efficiently inserted into the mitochondrial outer rather than the inner membrane in a membrane potential-independent manner. The carboxyl-terminal hydrophobic stretch is essential for the Bcl-2 protein to integrate into the outer membrane. Binding of the Bcl-2 protein to the mitochondria was not affected by pretreatment of the mitochondria with trypsin, indicating that the Bcl-2 protein does not require protease-sensitive mitochondrial surface components for its membrane insertion.

Animals↗

Size and localization of dystrophin molecule: immunoelectron microscopic and freeze etching studies of muscle plasma membranes of murine skeletal myofibers.

The ultrastructure and mode of existence of the dystrophin molecule and its relations to actin filaments were examined in murine skeletal myofibers. Electron microscopy of freeze-etched replicas of gold-labelled dystrophin molecules in quick-freeze, deep-etch, rotary-shadow preparations revealed rod-like structures 108.2 +/- 16.3 nm long and 3.1 +/- 1.5 nm thick. Some dystrophin molecules appeared to link their ends to form anastomosing networks; others were separate from each other. The dystrophin molecules were parallel or nearly parallel to the inner surface of the muscle plasma membrane. Double immuno-labelling transmission electron microscopy using N- and C-terminal dystrophin antibodies showed that the group mean distances of the N- and C-terminal signals from the muscle plasma membrane were 52.7 +/- 8.1 nm and 45.9 +/- 11.3 nm, respectively, which were not significantly different. Histograms of the distribution of the N- and C-terminal distances from the muscle plasma membrane had similar patterns with peaks 10-20 nm from the membrane. This was consistent with the findings of the mode of existence of dystrophin molecules seen in freeze-etched replicas. Finally, the dystrophin molecules were linked with the most peripheral sarcoplasmic actin like filaments, end to side as well as end to end.

Amino Acid Sequence↗