Biomedical subjects
A Inoue
Publications and source records attributed to A Inoue.
[Chronic eosinophilic pneumonia associated with high concentrations of interleukin-5, -6, and granulocyte colony stimulating factor in serum and in pleural fluid].
A 44-year-old Japanese man who had suffered from bronchial asthma since childhood was given the diagnosis of chronic eosinophilic pneumonia because of his symptoms, chest roentgenographic findings, and the results of a transbronchial lung biopsy. At the time of the onset of the disease, the pleural effusion contained 73% eosinophils. Symptoms were relieved and the laboratory findings returned forward normal after a short course of high-dose corticosteroids. The concentrations of IL-5, IL-6, and G-CSF in pleural fluid and in serum were very high; the concentrations of these cytokines were 3 times to 35 times higher in pleural fluid than in serum. In contrast, no IL-3 or GM-CSF was detected in any of these samples. The precise etiology of chronic eosinophilic pneumonia is still unclear, but this case suggests that inappropriate production of IL-5, IL-6 and G-CSF in the lung play a pivotal role in this disease. Inhibition of the production of these cytokines may be another therapeutic approach to this disease.
[A case of Goodpasture's syndrome with myeloperoxidase specific anti-neutrophil cytoplasmic autoantibody (MPO-ANCA) during chronic interstitial pneumonia].
A 55-year-old man was admitted to our hospital with of hemoptysis, progression of anemia and renal failure in February, 1996. Idiopathic interstitial pneumonia had been diagnosed and he had been followed at a regional hospital since 1988. On the third day after admission, he suffered from sudden and massive hemoptysis. Goodpasture's syndrome was diagnosed because anti-GBM antibody was detected in serum. A high titer of MPO-ANCA was also recognized simultaneously. Steroid pulse therapy, immunosuppressive therapy, and plasmapheresis were begun, but he died on the 28th hospital day because of severe hypoxemia and multi-organ failure. Histological examination after autopsy revealed crescentic glomerulonephritis with linear deposition of IgG in the glomerular capillary wall, and interstitial pneumonia accompanied by massive alveolar hemorrhage. It was suggested that in this patient, not only anti-GBM antibody but also circulating MPO-ANCA might have participated in the progression of the crescentic glomerulonephritis and alveolar hemorrhage observed in Goodpasture's syndrome.
Stimulation by retinoids of the natriuretic peptide system of osteoblastic MC3T3-E1 cells.
The effects were examined of treatment with retinoids of osteoblastic MC3T3-E1 cells on the natriuretic peptide system that promotes the differentiation of osteoblastic cells. Northern blot analysis revealed high levels of mRNA for the retinoid X receptor beta (RXR beta) and moderate levels of mRNAs for retinoic acid receptors alpha (RAR alpha) and gamma (RAR gamma). Exposure of MC3T3-E1 cells to 1 microM retinoid caused increases in the levels of C-type natriuretic peptide (CNP) and natriuretic peptide receptor-C (NPR-C). The activity of natriuretic peptide receptor-B (NPR-B) was unchanged after the addition of retinoid to the culture system. These results suggest that retinoids might influence the metabolism of osteoblastic cells through regulation of the natriuretic peptide system.
The simulated binding of (+/-)-2,3-dihydro-5,6-dimethoxy-2-[[1-(phenylmethyl)-4-piperidinyl]meth yl] -1H-inden-1-one hydrochloride (E2020) and related inhibitors to free and acylated acetylcholinesterases and corresponding structure-activity analyses.
The simulated binding profiles of acetylcholine, ACh, and the inhibitor (+/-)-2,3-dihydro-5,6- dimethoxy-2-[[1-(phenylmethyl)-4-piperidinyl]methyl]-1H-inden-1-on e hydrochloride (E2020), 1, and some of its analogs to acetylcholinesterase, AChE, were determined using full force field energetics and allowing complete conformational flexibility in both the ligand and receptor. A new mode of binding of ACh to AChE was found which involves the carboxyl oxygen of ACh interacting with Gly 118 and 119. Multiple modes of binding of 1 and some of its analogs were found which include alignment models observed in previous more restricted modeling studies. The key ligand-receptor interactions identified, and the corresponding energetics, are consistent on a relative basis, with observed binding constants for both the individual isomers of each of the inhibitors, as well as among the inhibitors themselves. The multiple modes of binding of 1 to AChE arises from small changes in binding at a single subsite and also from multiple subsite changes. Thus, an independent subsite model for ligand-receptor binding holds for some modes of binding, but not for others. A comparison of the simulated AChE-1 (and analog inhibitors) binding models to the receptor-independent 3D-QSARs previously developed for this class of inhibitors reveals extensive mutual consistency. The findings from these two modeling studies provides greater guidelines for inhibitor design than can be realized from either one. The combined docking and 3D-QSAR studies permit a detailed understanding of the SAR of more than 100 compound 1 analog inhibitors. A simple molecular recognition model can also be gleaned from the docking studies. A cylindrical "plug" (the inhibitor) having a large dipole moment must sterically fit into a cylindrical hole (the active site gorge of AChE), the lining of which also has a large dipole moment. Our simulations suggest that the dynamic "back door" to the active site of AChE does not form a large enough opening for sufficiently long time periods so as to be an effective entrance/exit pathway.
Magnetoresistance of Al90Y10 and Al90La10: Strong enhancement due to small crystalline precipitates in Al90La10.
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Electronic Structure of Icosahedral Alloys Studied by Ultrahigh Energy Resolution Photoemission Spectroscopy.
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Evidence that glucose metabolism is decreased in the cerebrum of aged female senescence-accelerated mouse; possible involvement of a low hexokinase activity.
d-Glucose metabolism in cerebral cells prepared from aged senescence-accelerated mouse (SAM), was investigated in consideration of a sex difference. The production of 14CO2 from 6-[14C]D-glucose was reduced in female senescence-accelerated-prone mouse (SAMP) 8, a prone substrain, in comparison with that in female senescence-accelerated-resistant mouse (SAMR) 2, a control substrain, whereas there was no difference in males. The 2-deoxy-D-glucose uptake into cerebral cells from female SAMP8 was also lower than that of control mice. But, the 3-O-methyl-D-glucose uptake in SAMP8 was higher than that of SAMR2, suggesting that the low hexokinase activity was involved in the decreased glucose metabolism in cerebrum of SAMP8 females irrespective of glucose transporter. This possibility was supported by the finding that the contents of glucose 6-phosphate produced from glucose added to cerebral cells from SAMP8 was lower than that in ICR mice.
Molecular cloning of a RNA binding protein, S1-1.
S1 proteins A-D constitute a nuclear protein family that are liberated rapidly in a set from chromatin by mild digestion with a DNA or RNA hydrolyzing enzyme. With an anti-S1-protein B antiserum that reacted with B2, C1 and D1, a cDNA clone, pS1-1, was obtained, which encoded a protein of 852 amino acids. The S1-1 protein, encoded within the cells by a mRNA of 3480 nt, was a novel protein and could be distinguished from the S1 proteins B, C and D by their amino acid sequences. The S-1-1 protein synthesized by in vitro translation bound to RNA homopolymers, with a preference for G and U polyribonucleotides and little for poly(A). The protein contained two tandem RNP motifs and several intriguing sequences, such as a novel repeat of five octamers with a consensus sequence DP-S(Q/G)YYY and a potentially perfect amphipathic alpha-helix of five turns with basic and acidic amino acids positioned in an ordered way. The two RNP motif sequences were similar, although homologies were low, to the RNP motif sequences of yeast NSR1 protein, animal nucleolins, Drosophila hnRNP Al and tobacco chloroplast RNP precursor protein, suggesting a functional uniqueness of the S1-1 protein in RNA metabolism and also the evolution of its RNP motif structure before plants and animals diverged. These results indicate that the S1-1 protein encoded by the cDNA is a new class of RNA binding protein.
Inhibition of proliferation of chondrocytes by specific receptors in response to retinoids.
All-trans retinoic acid inhibited the proliferation of chondrocytes derived from rat xiphoid cartilage when added to the culture medium at 10(-10)-10(-8) M. Proliferation of mouse clonal osteoblastic cells was also inhibited to a significant extent by all-trans retinoic acid. However, no such inhibitory effects on rat smooth muscle cells and human fibroblasts were observed. Flow cytometric analyses of chondrocytes labeled with propidium iodide revealed that all-trans retinoic acid arrested chondrocytes at the G1 phase of the cell cycle. Since 9-cis retinoic acid, which is synthesized enzymatically from all-trans retinoic acid, also inhibited the proliferation of chondrocytes, we investigated the subtypes of retinoic acid receptors in chondrocytes. Northern blot analysis revealed high levels of mRNA for retinoid X receptor alpha (RXR alpha), moderate levels of mRNA for retinoic acid receptor gamma (RAR gamma), and low levels of mRNA for RAR alpha and RXR beta. The mRNA for RAR beta and RXR gamma were not detected. These results suggest that retinoids are associated with the inhibition of proliferation of chondrocytes via both families of nuclear receptors, namely, RARs and RXRs.
Stimulation by C-type natriuretic peptide of the differentiation of clonal osteoblastic MC3T3-E1 cells.
We examined the effects of C-type natriuretic peptide (CNP) and B-type natriuretic peptide receptor (NPR-B) system, which stimulates the intracellular production of cGMP, on osteoblastic differentiation using clonal murine calvarial MC3T3-E1 cells. CNP-like immunoreactivity was detected in the conditioned medium and in lysates of MC3T3-E1 cells. Exposure of cells to CNP caused an increase in the intracellular production of cGMP and the increase was dose-dependent, while ANP had no effect. These results imply that CNP regulates osteoblastic metabolism via NPR-B in an autocrine manner. Northern blot analysis revealed that treatment of MC3T3-E1 cells with CNP increased the steady-state levels of mRNAs for type-I collagen, cellular alkaline phosphatase (ALPase), and osteocalcin, which are well known as markers of osteoblastic differentiation. Our observations suggest the possibility that CNP functions as a local regulator of osteoblastic differentiation, acting via a cGMP-mediated pathway.
Cloning, sequencing and transcriptional regulation of the draT and draG genes of Azospirillum lipoferum FS.
From Azospirillum lipoferum (Al) FS, a nitrogen-fixing bacterium isolated from the rhizosphere of rice, we cloned and sequenced draT, encoding dinitrogenase reductase ADP-ribosyltransferase, and draG, encoding dinitrogenase reductase-activating glycohydrolase. The nucleotide sequences of draTG showed extensive similarity to the same genes from Azospirillum brasilense, Rhodospirillum rubrum and Rhodobacter capsulatus, and they are assumed to be co-transcribed as a single operon. When this draTG operon was introduced into Klebsiella oxytoca, this organism acquired the ability to respond to extracellular NH(+4) ions with reversible inhibition of nitrogenase activity, similar to that seen in Al FS. We constructed a plasmid containing a draT::lacZ gene fusion and found that beta-galactosidase activity was detected under microaerobic conditions, regardless of NH(+4) concentration, but not under aerobic conditions. This indicates that the transcription of draTG responds to the level of oxygen, but not to that of NH(+4) ions.
Morphological and biochemical evidence for apoptosis in the terminal hypertrophic chondrocytes of the growth plate.
The purpose of this study was to investigate the mechanism of cell death in chondrocytes of the growth plate. In the degenerative chondrocyte zone of the growth plate, apoptotic chondrocytes were defeated by the in situ nick end labelling method, by DNA analysis in agarose gel, and by electron microscopy. The results of the in situ nick end labelling method and the occurrence of a ladder pattern of DNA in agarose gel analysis indicated the activation of endogenous endonucleases, resulting in DNA fragmentation. Electron micrographs showed the early morphological changes associated with apoptosis. This report presents both morphological and biochemical evidence for apoptosis in the terminal hypertrophic chondrocytes of the growth plate. These data suggest that apoptosis of degenerative chondrocytes may play an important role in the control of normal and pathological endochondral ossification.
Differentiation of smooth muscle cells from undifferentiated cells of chicken gizzard occurs on the layer of fibroblast-like cells.
We examined the in vitro differentiation of smooth muscle cells from undifferentiated cells of embryonic chicken gizzard. We used the gizzard from 7-day-old embryos (Hamburger and Hamilton's stage 26-28) as a source of culture, since the expression of myosin is extremely low, and the gizzard consists of round cells, which are not stained by anti-smooth muscle myosin antiserum. When the dissections of the gizzard were cultured, they attached to the dish, and fibroblast-like cells migrated within 4 days after the culture. Then round cells migrated from the transplants over the layer of fibroblast-like cells. At 12-14 days after the culture, smooth muscle cells, which were ribbon-shaped and stained by anti-smooth muscle myosin antiserum, appeared in the layer of round cells. The dissected transplant itself was not stained by anti-smooth muscle myosin antiserum even after being cultured for 15 days. We concluded then that the smooth muscle cells were differentiated from the round cells, which spread on the layer of the fibroblast-like cells. We also observed the differentiation of smooth muscle cells when the cells separated from 7-day-old embryo cultured on the layer of cloned fibroblast cells. We suggest that the fibroblast-like cell may play an important role in the differentiation of smooth muscle cells.
Suppression of Theiler's murine encephalomyelitis virus induced demyelinating disease by administration of gangliosides.
Intracerebral (i.c.) inoculation of susceptible strains of mice with Theiler's murine encephalomyelitis virus (TMEV) results in immune-mediated demyelinating disease. Gangliosides are membrane components of essentially all eukaryotic cells and are abundant in plasma membranes. Endogenous gangliosides have been implicated in cell recognition, cell adhesion, cell differentiation and neurite outgrowth. We studied the effect of gangliosides on TMEV-induced demyelinating disease (TMEV- IDD). We injected TMEV intracerebrally into susceptible SJL/J mice and induced TMEV-IDD. Gangliosides were injected subcutaneously and examined for various immunological indicators. The results show that when gangliosides were administered in the effector phase, TMEV-IDD was suppressed both clinically and histologically. Cellular immunity such as delayed-type hypersensitivity, and the proliferative response of T cells against TMEV and mitogens were decreased, and only in this group anti-TMEV IgG2a antibody was not detected. Taken together, these data suggest that administration of gangliosides suppressed the function of pathogenic Th1 cells and suppressed TMEV-IDD. Additionally, this study proposes the possibility of a new therapy in multiple sclerosis.
Isolating and characterizing deep-sea marine microorganisms.
We have isolated several microorganisms that are adapted to living in the extremes of the deep-sea environment. They include barophilic bacteria, which are able to grow at high hydrostatic pressure, but that are unable to grow at atmospheric pressure, and organic-solvent-tolerant bacteria, which are able to grow in the presence of toxic organic solvents such as toluene or benzene. In this review, we describe how to isolate such extremophiles, and we outline the characteristics of several strains that have been recovered from the deep-sea environment.
5-HT3 receptor blocking properties of the antiparkinsonian agent, talipexole.
1. Talipexole showed moderate displacement activity of 3H-GR 65630 binding to 5-HT3 receptors in both rat cortical and intestinal membrane fractions with Ki values of 0.35 microM and 0.22 microM, respectively. 2. Bromocriptine failed to displace the binding activity in either experimental system even at a concentration of 10 microM. 3. Both talipexole and tropisetron were found to significantly inhibit 5-HT3 receptor-mediated effects of 5-HT in isolated guinea-pig ileum or atrium; however, the effect of talipexole was weaker than that of tropisetron. 4. Bromocriptine, in contrast, had no antagonistic effects on 5-HT3 receptor-mediated activity in guinea-pig ileum or atrium. 5. It was concluded that talipexole might act as an antagonist on 5-HT3 receptors in both brain and intestinal tissues.
The rationale for E2020 as a potent acetylcholinesterase inhibitor.
The phase III drug-candidate, E2020, developed for treatment of Alzheimer's disease, and possibly other demenitas, and its analogues have been the focus of extensive molecular pharmacological and structural studies. The potency and selectivity of E2020 as an inhibitor of acetylcholinesterase, AChE, in the brain is established. A combination of molecular modeling and QSAR studies have been used throughout the evolution of the AChE inhibitor program leading to the benzylpiperidine series, and, ultimately, E2020. QSAR studies have identified requirements of optimize inhibition activity as a function of substituent choice on both the indanone and benzyl rings in the E2020 class of inhibitors. A combination of X-ray crystal structure studies of E2020 isomers and the molecular shape analysis, MSA, of E2020 and its analogues has led to a postulated active conformation, and molecular shape, for these AChE inhibitors. The active molecular shape corresponds to a high degree of shape similarity between the two E2020 isomers which, in turn, is consistent with the observed high inhibition potencies of both of these compounds. Intermolecular docking studies were carried out for E2020 and some analogues with the crystal structure of AChE when it became available. The docking simulations involving E2020 analogues suggest these inhibitors do not bind at the acetylcholine, ACh, active site, but rather at the most narrow location of the long channel leading to the active site. Intermolecular binding geometries are consistent with the postulated active conformations derived from structure-activity (receptor geometry independent) information.