Tanaka H

References (42)

Title : Acanthopanax senticosus ameliorates steatohepatitis through HNF4 alpha pathway activation in mice - Kawano_2024_Sci.Rep_14_110
Author(s) : Kawano Y , Tanaka M , Satoh Y , Sugino S , Suzuki J , Fujishima M , Okumura E , Takekoshi H , Uehara O , Sugita S , Abiko Y , Tomonari T , Tanaka H , Takeda H , Takayama T
Ref : Sci Rep , 14 :110 , 2024
Abstract : Non-alcoholic fatty liver disease is a common liver disease worldwide, and is associated with dysregulation of lipid metabolism, leading to inflammation and fibrosis. Acanthopanax senticosus Harms (ASH) is widely used in traditional medicine as an adaptogen food. We examined the effect of ASH on steatohepatitis using a high-fat diet mouse model. Mice were fed a choline-deficient, L-amino acid-defined, high-fat diet with ASH extract (ASHE). After 6 weeks, liver RNA transcriptome sequencing (RNA-Seq) was performed, followed by Ingenuity Pathway Analysis (IPA). Our findings revealed that mice fed a high-fat diet with 5% ASHE exhibited significantly reduced liver steatosis. These mice also demonstrated alleviated inflammation and reduced fibrosis in the liver. IPA of RNA-Seq indicated that hepatocyte nuclear factor 4 alpha (HNF4 alpha), a transcription factor, was the activated upstream regulator (P-value 0.00155, z score = 2.413) in the liver of ASHE-fed mice. Adenosine triphosphate binding cassette transporter 8 and carboxylesterase 2, downstream targets of HNF4 alpha pathway, were upregulated. Finally, ASHE-treated HepG2 cells exposed to palmitate exhibited significantly decreased lipid droplet contents. Our study provides that ASHE can activate HNF4 alpha pathway and promote fat secretion from hepatocytes, thereby serving as a prophylactic treatment for steatohepatitis in mice.
ESTHER : Kawano_2024_Sci.Rep_14_110
PubMedSearch : Kawano_2024_Sci.Rep_14_110
PubMedID: 38167633

Title : Whole-body insulin resistance and energy expenditure indices, serum lipids, and skeletal muscle metabolome in a state of lipoprotein lipase overexpression - Nishida_2021_Metabolomics_17_26
Author(s) : Nishida Y , Nishijima K , Yamada Y , Tanaka H , Matsumoto A , Fan J , Uda Y , Tomatsu H , Yamamoto H , Kami K , Kitajima S , Tanaka K
Ref : Metabolomics , 17 :26 , 2021
Abstract : INTRODUCTION: Overexpression of lipoprotein lipase (LPL) protects against high-fat-diet (HFD)-induced obesity and insulin resistance in transgenic rabbits; however, the molecular mechanisms remain unclear. Skeletal muscle is a major organ responsible for insulin-stimulated glucose uptake and energy expenditure. OBJECTIVES: The main purpose of the current study was to examine the effects of the overexpression of LPL on the skeletal muscle metabolomic profiles to test our hypothesis that the mitochondrial oxidative metabolism would be activated in the skeletal muscle of LPL transgenic rabbits and that the higher mitochondrial oxidative metabolism activity would confer better phenotypic metabolic outcomes. METHODS: Under a HFD, insulin resistance index was measured using the intravenous glucose tolerance test, and total energy expenditure (TEE) was measured by doubly-labeled water in control and LPL transgenic rabbits (n = 12, each group). Serum lipids, such as triglycerides and free fatty acid, were also measured. The skeletal muscle metabolite profile was analyzed using capillary electrophoresis time-of flight mass spectrometry in the two groups (n = 9, each group). A metabolite set enrichment analysis (MSEA) with muscle metabolites and a false discovery rate q < 0.2 was performed to identify significantly different metabolic pathways between the 2 groups. RESULTS: The triglycerides and free fatty acid levels and insulin resistance index were lower, whereas the TEE was higher in the LPL transgenic rabbits than in the control rabbits. Among 165 metabolites detected, the levels of 37 muscle metabolites were significantly different between the 2 groups after false discovery rate correction (q < 0.2). The MSEA revealed that the TCA cycle and proteinogenic amino acid metabolism pathways were significantly different between the 2 groups (P < 0.05). In the MSEA, all four selected metabolites for the TCA cycle (2-oxoglutaric acid, citric acid, malic acid, fumaric acid), as well as eight selected metabolites for proteinogenic amino acid metabolism (asparagine, proline, methionine, phenylalanine, histidine, arginine, leucine, isoleucine) were consistently increased in the transgenic rabbits compared with control rabbits, suggesting that these two metabolic pathways were activated in the transgenic rabbits. Some of the selected metabolites, such as citric acid and methionine, were significantly associated with serum lipids and insulin resistance (P < 0.05). CONCLUSION: The current results suggest that the overexpression of LPL may lead to increased activities of TCA cycle and proteinogenic amino acid metabolism pathways in the skeletal muscle, and these enhancements may play an important role in the biological mechanisms underlying the anti-obesity/anti-diabetes features of LPL overexpression.
ESTHER : Nishida_2021_Metabolomics_17_26
PubMedSearch : Nishida_2021_Metabolomics_17_26
PubMedID: 33594546

Title : Visualization of Synchronous or Asynchronous Release of Single Synaptic Vesicle in Active-Zone-Like Membrane Formed on Neuroligin-Coated Glass Surface - Funahashi_2018_Front.Cell.Neurosci_12_140
Author(s) : Funahashi J , Tanaka H , Hirano T
Ref : Front Cell Neurosci , 12 :140 , 2018
Abstract : Fast repetitive synaptic transmission depends on efficient exocytosis and retrieval of synaptic vesicles around a presynaptic active zone. However, the functional organization of an active zone and regulatory mechanisms of exocytosis, endocytosis and reconstruction of release-competent synaptic vesicles have not been fully elucidated. By developing a novel visualization method, we attempted to identify the location of exocytosis of a single synaptic vesicle within an active zone and examined movement of synaptic vesicle protein synaptophysin (Syp) after exocytosis. Using cultured hippocampal neurons, we induced formation of active-zone-like membranes (AZLMs) directly adjacent and parallel to a glass surface coated with neuroligin, and imaged Syp fused to super-ecliptic pHluorin (Syp-SEP) after its translocation to the plasma membrane from a synaptic vesicle using total internal reflection fluorescence microscopy (TIRFM). An AZLM showed characteristic molecular and functional properties of a presynaptic active zone. It contained active zone proteins, cytomatrix at the active zone-associated structural protein (CAST), Bassoon, Piccolo, Munc13 and RIM, and showed an increase in intracellular Ca(2+) concentration upon electrical stimulation. In addition, single-pulse stimulation sometimes induced a transient increase of Syp-SEP signal followed by lateral spread in an AZLM, which was considered to reflect an exocytosis event of a single synaptic vesicle. The diffusion coefficient of Syp-SEP on the presynaptic plasma membrane after the membrane fusion was estimated to be 0.17-0.19 mum(2)/s, suggesting that Syp-SEP diffused without significant obstruction. Synchronous exocytosis just after the electrical stimulation tended to occur at multiple restricted sites within an AZLM, whereas locations of asynchronous release occurring later after the stimulation tended to be more scattered.
ESTHER : Funahashi_2018_Front.Cell.Neurosci_12_140
PubMedSearch : Funahashi_2018_Front.Cell.Neurosci_12_140
PubMedID: 29875634

Title : Direct reprogramming of fibroblasts into skeletal muscle progenitor cells by transcription factors enriched in undifferentiated subpopulation of satellite cells - Ito_2017_Sci.Rep_7_8097
Author(s) : Ito N , Kii I , Shimizu N , Tanaka H , Shin'ichi T
Ref : Sci Rep , 7 :8097 , 2017
Abstract : Satellite cells comprise a functionally heterogeneous population of stem cells in skeletal muscle. Separation of an undifferentiated subpopulation and elucidation of its molecular background are necessary to identify the reprogramming factors to induce skeletal muscle progenitor cells. In this study, we found that intracellular esterase activity distinguishes a subpopulation of cultured satellite cells with high stemness using esterase-sensitive cell staining reagent, calcein-AM. Gene expression analysis of this subpopulation revealed that defined combinations of transcription factors (Pax3, Mef2b, and Pitx1 or Pax7, Mef2b, and Pitx1 in embryonic fibroblasts, and Pax7, Mef2b and MyoD in adult fibroblasts) reprogrammed fibroblasts into skeletal muscle progenitor cells. These reprogrammed cells formed Dystrophin-positive mature muscle fibers when transplanted into a mouse model of Duchenne muscular dystrophy. These results highlight the new marker for heterogenous population of cultured satellite cells, potential therapeutic approaches and cell sources for degenerative muscle diseases.
ESTHER : Ito_2017_Sci.Rep_7_8097
PubMedSearch : Ito_2017_Sci.Rep_7_8097
PubMedID: 28808339

Title : A new metabolism-related index correlates with the degree of liver fibrosis in hepatitis C virus-positive patients - Enomoto_2015_Gastroenterol.Res.Pract_2015_926169
Author(s) : Enomoto H , Aizawa N , Nakamura H , Takata R , Sakai Y , Iwata Y , Tanaka H , Ikeda N , Aoki T , Hasegawa K , Yoh K , Hashimoto K , Ishii A , Takashima T , Saito M , Imanishi H , Iijima H , Nishiguchi S
Ref : Gastroenterol Res Pract , 2015 :926169 , 2015
Abstract : Background. Only a few biomarkers based on metabolic parameters for evaluating liver fibrosis have been reported. The aim of this study was to investigate the relevance of an index obtained from three metabolic variables (glycated albumin: GA, glycated hemoglobin: HbA1c, and branched-chain amino acids to tyrosine ratio: BTR) to the degree of liver fibrosis in hepatitis C virus virus- (HCV-) positive patients. Methods. A total of 394 HCV-positive patients were assessed based on the values of a new index (GA/HbA1c/BTR). The index findings were used to investigate the relationship with the degree of liver fibrosis. Results. The new index showed an association with the stage of fibrosis (METAVIR scores: F0-1: 0.42 +/- 0.10, F2: 0.48 +/- 0.15, F3: 0.56 +/- 0.22, and F4: 0.71 +/- 0.30). The index was negatively correlated with three variables of liver function: the prothrombin time percentage (P < 0.0001), albumin level (P < 0.0001), and cholinesterase level (P < 0.0001). The new index showed a higher correlation related to liver function than FIB-4 and the APRI did. In addition, the index showed a higher AUROC value than that of FIB-4 and the APRI for prediction of liver cirrhosis. Conclusion. The new metabolism-related index, GA/HbA1c/BTR value, is shown to relate to the degree of liver fibrosis in HCV-positive patients.
ESTHER : Enomoto_2015_Gastroenterol.Res.Pract_2015_926169
PubMedSearch : Enomoto_2015_Gastroenterol.Res.Pract_2015_926169
PubMedID: 25861264

Title : An Increased Ratio of Glycated Albumin to HbA1c Is Associated with the Degree of Liver Fibrosis in Hepatitis B Virus-Positive Patients - Enomoto_2014_Gastroenterol.Res.Pract_2014_351396
Author(s) : Enomoto H , Aizawa N , Nakamura H , Sakai Y , Iwata Y , Tanaka H , Ikeda N , Aoki T , Yuri Y , Yoh K , Hashimoto K , Ishii A , Takashima T , Iwata K , Saito M , Imanishi H , Iijima H , Nishiguchi S
Ref : Gastroenterol Res Pract , 2014 :351396 , 2014
Abstract : Background. In hepatitis B virus- (HBV-) positive patients, the relationship between the metabolic variables and histological degree of liver fibrosis has been poorly investigated. Methods. A total of 176 HBV-positive patients were assessed in whom the ratios of glycated albumin-to-glycated hemoglobin (GA/HbA1c) were calculated in order to investigate the relationship with the degree of liver fibrosis. Results. The GA/HbA1c ratio increased in association with the severity of fibrosis (METAVIR scores: F0-1: 2.61 +/- 0.24, F2: 2.65 +/- 0.24, F3: 2.74 +/- 0.38, and F4: 2.91 +/- 0.63). The GA/HbA1c ratios were inversely correlated with four variables of liver function: the prothrombin time (PT) percentage (P < 0.0001), platelet count (P < 0.0001), albumin value (P < 0.0001), and cholinesterase value (P < 0.0001). The GA/HbA1c ratio was positively correlated with two well-known markers of liver fibrosis, FIB-4 (P < 0.0001) and the AST-to-platelet ratio index (APRI) (P < 0.0001). Furthermore, the GA/HbA1c showed better correlations with two variables of liver function (PT percentage and cholinesterase value) than did FIB-4 and with all four variables than did the APRI. Conclusion. The GA/HbA1c ratio is associated with the degree of liver fibrosis in HBV-positive patients.
ESTHER : Enomoto_2014_Gastroenterol.Res.Pract_2014_351396
PubMedSearch : Enomoto_2014_Gastroenterol.Res.Pract_2014_351396
PubMedID: 24693282

Title : Stepwise enhancement of catalytic performance of haloalkane dehalogenase LinB towards beta-hexachlorocyclohexane - Moriuchi_2014_AMB.Express_4_72
Author(s) : Moriuchi R , Tanaka H , Nikawadori Y , Ishitsuka M , Ito M , Ohtsubo Y , Tsuda M , Damborsky J , Prokop Z , Nagata Y
Ref : AMB Express , 4 :72 , 2014
Abstract : Two haloalkane dehalogenases, LinBUT and LinBMI, each with 296 amino acid residues, exhibit only seven amino acid residue differences between them, but LinBMI's catalytic performance towards beta-hexachlorocyclohexane (beta-HCH) is considerably higher than LinBUT's. To elucidate the molecular basis governing this difference, intermediate mutants between LinBUT and LinBMI were constructed and kinetically characterized. The activities of LinBUT-based mutants gradually increased by cumulative mutations into LinBUT, and the effects of the individual amino acid substitutions depended on combination with other mutations. These results indicated that LinBUT's beta-HCH degradation activity can be enhanced in a stepwise manner by the accumulation of point mutations.
ESTHER : Moriuchi_2014_AMB.Express_4_72
PubMedSearch : Moriuchi_2014_AMB.Express_4_72
PubMedID: 25401073

Title : Endothelial lipase modulates pressure overload-induced heart failure through alternative pathway for fatty acid uptake - Nakajima_2013_Hypertension_61_1002
Author(s) : Nakajima H , Ishida T , Satomi-Kobayashi S , Mori K , Hara T , Sasaki N , Yasuda T , Toh R , Tanaka H , Kawai H , Hirata K
Ref : Hypertension , 61 :1002 , 2013
Abstract : Lipoprotein lipase has been considered as the only enzyme capable of generating lipid-derived fatty acids for cardiac energy. Endothelial lipase is another member of the triglyceride lipase family and hydrolyzes high-density lipoproteins. Although endothelial lipase is expressed in the heart, its function remains unclear. We assessed the role of endothelial lipase in the genesis of heart failure. Pressure overload-induced cardiac hypertrophy was generated in endothelial lipase(-/-) and wild-type mice by ascending aortic banding. Endothelial lipase expression in cardiac tissues was markedly elevated in the early phase of cardiac hypertrophy in wild-type mice, whereas lipoprotein lipase expression was significantly reduced. Endothelial lipase(-/-) mice showed more severe systolic dysfunction with left-ventricular dilatation compared with wild-type mice in response to pressure overload. The expression of mitochondrial fatty acid oxidation-related genes, such as carnitine palmitoyltransferase-1 and medium-chain acyl coenzyme A dehydrogenase, was significantly lower in the heart of endothelial lipase(-/-) mice than in wild-type mice. Also, endothelial lipase(-/-) mice had lower myocardial adenosine triphosphate levels than wild-type mice after aortic banding. In cultured cardiomyocytes, endothelial lipase was upregulated by inflammatory stimuli, whereas lipoprotein lipase was downregulated. Endothelial lipase-overexpression in cardiomyocytes resulted in an upregulation of fatty acid oxidation-related enzymes and intracellular adenosine triphosphate accumulation in the presence of high-density lipoprotein. Endothelial lipase may act as an alternative candidate to provide fatty acids to the heart and regulate cardiac function. This effect seemed relevant particularly in the diseased heart, where lipoprotein lipase action is downregulated.
ESTHER : Nakajima_2013_Hypertension_61_1002
PubMedSearch : Nakajima_2013_Hypertension_61_1002
PubMedID: 23460280

Title : Donepezil Abolishes Anticholinergic Activity in a Patient with Amnesia - Konishi_2012_Pharmacology_91_86
Author(s) : Konishi K , Hori K , Tomioka H , Minegishi G , Tani M , Tanaka H , Akita R , Yokoyama S , Oshio T , Hachisu M
Ref : Pharmacology , 91 :86 , 2012
Abstract : We report the case of a 74-year-old woman who presented with amnesia and positive serum anticholinergic activity (SAA), which disappeared after treatment with the cholinesterase inhibitor donepezil for 1 year. Her only other regular medications were topical glaucoma preparations. We suggest that mental stress, mild cognitive impairment and Alzheimer's disease pathology combined to generate SAA in this patient. We also consider that SAA may have subsequently become negative because of upregulation of acetylcholine production by donepezil, and because the patient's other medications and physical condition (including glaucoma) remained unchanged during the 1-year period.
ESTHER : Konishi_2012_Pharmacology_91_86
PubMedSearch : Konishi_2012_Pharmacology_91_86
PubMedID: 23258422

Title : Synthesis of phenserine analogues and evaluation of their cholinesterase inhibitory activities - Shinada_2012_Bioorg.Med.Chem_20_4901
Author(s) : Shinada M , Narumi F , Osada Y , Matsumoto K , Yoshida T , Higuchi K , Kawasaki T , Tanaka H , Satoh M
Ref : Bioorganic & Medicinal Chemistry , 20 :4901 , 2012
Abstract : Phenserine is a potentially attractive drug for Alzheimer's disease. In order to further expand SAR study for inhibitions of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE), the methyl group at the 3a-position of phenserine was replaced with an alkyl or alkenyl group, and its phenylcarbamoyl moiety was substituted at the o- or p-position. The synthetic methodology for these phenserine analogues includes the efficient cascade reactions for introduction of the 3a-substituent and assembly of the quaternary carbon center followed by reductive cyclization to the key pyrroloindoline structure. The bulkiness of the substituent at 3a-position of phenserine derivatives tends to reduce the inhibitory effect on AChE activity in the following order: methyl > ethyl > vinyl > propyl approximately allyl > reverse-prenyl groups. Among the series synthesized, the 3a-ethyl derivative demonstrated the highest AChE selectivity. In construct, the 3a-reverse-prenyl derivative indicated modest BuChE selectivity.
ESTHER : Shinada_2012_Bioorg.Med.Chem_20_4901
PubMedSearch : Shinada_2012_Bioorg.Med.Chem_20_4901
PubMedID: 22831800

Title : Higher-order architecture of cell adhesion mediated by polymorphic synaptic adhesion molecules neurexin and neuroligin. - Tanaka_2012_Cell.Rep_2_101
Author(s) : Tanaka H , Miyazaki N , Matoba K , Nogi T , Iwasaki K , Takagi J
Ref : Cell Rep , 2 :101 , 2012
Abstract : Polymorphic adhesion molecules neurexin and neuroligin (NL) mediate asymmetric trans-synaptic adhesion, which is crucial for synapse development and function. It is not known whether or how individual synapse function is controlled by the interactions between variants and isoforms of these molecules with differing ectodomain regions. At a physiological concentration of Ca(2+), the ectodomain complex of neurexin-1 beta isoform (Nrx1beta) and NL1 spontaneously assembled into crystals of a lateral sheet-like superstructure topologically compatible with transcellular adhesion. Correlative light-electron microscopy confirmed extracellular sheet formation at the junctions between Nrx1beta- and NL1-expressing non-neuronal cells, mimicking the close, parallel synaptic membrane apposition. The same NL1-expressing cells, however, did not form this higher-order architecture with cells expressing the much longer neurexin-1 +/- isoform, suggesting a functional discrimination mechanism between synaptic contacts made by different isoforms of neurexin variants.
ESTHER : Tanaka_2012_Cell.Rep_2_101
PubMedSearch : Tanaka_2012_Cell.Rep_2_101
PubMedID: 22840401
Gene_locus related to this paper: ratno-1neur

Title : Structural basis for variant-specific neuroligin-binding by alpha-neurexin - Tanaka_2011_PLoS.One_6_e19411
Author(s) : Tanaka H , Nogi T , Yasui N , Iwasaki K , Takagi J
Ref : PLoS ONE , 6 :e19411 , 2011
Abstract : Neurexins (Nrxs) are presynaptic membrane proteins with a single membrane-spanning domain that mediate asymmetric trans-synaptic cell adhesion by binding to their postsynaptic receptor neuroligins. alpha-Nrx has a large extracellular region comprised of multiple copies of laminin, neurexin, sex-hormone-binding globulin (LNS) domains and epidermal growth factor (EGF) modules, while that of beta-Nrx has but a single LNS domain. It has long been known that the larger alpha-Nrx and the shorter beta-Nrx show distinct binding behaviors toward different isoforms/variants of neuroligins, although the underlying mechanism has yet to be elucidated. Here, we describe the crystal structure of a fragment corresponding to the C-terminal one-third of the Nrx1alpha ectodomain, consisting of LNS5-EGF3-LNS6. The 2.3 A-resolution structure revealed the presence of a domain configuration that was rigidified by inter-domain contacts, as opposed to the more common flexible "beads-on-a-string" arrangement. Although the neuroligin-binding site on the LNS6 domain was completely exposed, the location of the alpha-Nrx specific LNS5-EGF3 segment proved incompatible with the loop segment inserted in the B+ neuroligin variant, which explains the variant-specific neuroligin recognition capability observed in alpha-Nrx. This, combined with a low-resolution molecular envelope obtained by a single particle reconstruction performed on negatively stained full-length Nrx1alpha sample, allowed us to derive a structural model of the alpha-Nrx ectodomain. This model will help us understand not only how the large alpha-Nrx ectodomain is accommodated in the synaptic cleft, but also how the trans-synaptic adhesion mediated by alpha- and beta-Nrxs could differentially affect synaptic structure and function.
ESTHER : Tanaka_2011_PLoS.One_6_e19411
PubMedSearch : Tanaka_2011_PLoS.One_6_e19411
PubMedID: 21552542

Title : Targeted deletion of endothelial lipase increases HDL particles with anti-inflammatory properties both in vitro and in vivo - Hara_2011_J.Lipid.Res_52_57
Author(s) : Hara T , Ishida T , Kojima Y , Tanaka H , Yasuda T , Shinohara M , Toh R , Hirata K
Ref : J Lipid Res , 52 :57 , 2011
Abstract : Previous studies have shown that targeted deletion of endothelial lipase (EL) markedly increases the plasma high density lipoprotein cholesterol (HDL-C) level in mice. However, little is known about the functional quality of HDL particles after EL inhibition. Therefore, the present study assessed the functional quality of HDL isolated from EL(-/-) and wild-type (WT) mice. Anti-inflammatory functions of HDL from EL(-/-) and WT mice were evaluated by in vitro assays. The HDL functions such as PON-1 or PAF-AH activities, inhibition of cytokine-induced vascular cell adhesion molecule-1 expression, inhibition of LDL oxidation, and the ability of cholesterol efflux were similar in HDL isolated from WT and EL(-/-) mice. In contrast, the lipopolysaccharide-neutralizing capacity of HDL was significantly higher in EL(-/-) mice than that in WT mice. To evaluate the anti-inflammatory actions of HDL in vivo, lipopolysaccharide-induced systemic inflammation was generated in these mice. EL(-/-) mice showed higher survival rate and lower expression of inflammatory markers than WT mice. Intravenous administration of HDL isolated from EL(-/-) mice significantly improved the mortality after lipopolysaccharide injection in WT mice. In conclusion, targeted disruption of EL increased HDL particles with preserved anti-inflammatory and anti-atherosclerotic functions. Thus, EL inhibition would be a useful strategy to raise 'good' cholesterol in the plasma.
ESTHER : Hara_2011_J.Lipid.Res_52_57
PubMedSearch : Hara_2011_J.Lipid.Res_52_57
PubMedID: 20926433

Title : Function of phenylalanine 259 and threonine 314 within the substrate binding pocket of the juvenile hormone esterase of Manduca sexta - Kamita_2010_Biochemistry_49_3733
Author(s) : Kamita SG , Wogulis MD , Law CS , Morisseau C , Tanaka H , Huang H , Wilson DK , Hammock BD
Ref : Biochemistry , 49 :3733 , 2010
Abstract : Juvenile hormone (JH) is a key insect developmental hormone that is found at low nanomolar levels in larval insects. The methyl ester of JH is hydrolyzed in many insects by an esterase that shows high specificity for JH. We have previously determined a crystal structure of the JH esterase (JHE) of the tobacco hornworm Manduca sexta (MsJHE) [Wogulis, M., Wheelock, C. E., Kamita, S. G., Hinton, A. C., Whetstone, P. A., Hammock, B. D., and Wilson, D. K. (2006) Biochemistry 45, 4045-4057]. Our molecular modeling indicates that JH fits very tightly within the substrate binding pocket of MsJHE. This tight fit places two noncatalytic amino acid residues, Phe-259 and Thr-314, within the appropriate distance and geometry to potentially interact with the alpha,beta-unsaturated ester and epoxide, respectively, of JH. These residues are highly conserved in numerous biologically active JHEs. Kinetic analyses of mutants of Phe-259 or Thr-314 indicate that these residues contribute to the low K(M) that MsJHE shows for JH. This low K(M), however, comes at the cost of reduced substrate turnover. Neither nucleophilic attack of the resonance-stabilized ester by the catalytic serine nor the availability of a water molecule for attack of the acyl-enzyme intermediate appears to be a rate-determining step in the hydrolysis of JH by MsJHE. We hypothesize that the release of the JH acid metabolite from the substrate binding pocket limits the catalytic cycle. Our findings also demonstrate that chemical bond strength does not necessarily correlate with how reactive the bond will be to metabolism.
ESTHER : Kamita_2010_Biochemistry_49_3733
PubMedSearch : Kamita_2010_Biochemistry_49_3733
PubMedID: 20307057

Title : DNA methylation of the promoter of soluble epoxide hydrolase silences its expression by an SP-1-dependent mechanism - Zhang_2010_Biochim.Biophys.Acta_1799_659
Author(s) : Zhang D , Ai D , Tanaka H , Hammock BD , Zhu Y
Ref : Biochimica & Biophysica Acta , 1799 :659 , 2010
Abstract : Epoxyeicosatrienoic acids, derived from arachidonic acid, function as antihypertensive and antihypertrophic mediators in the cardiovascular system. They are hydrolyzed by soluble epoxide hydrolase (sEH). Pharmacological inhibition of sEH increases the level of epoxyeicosatrienoic acids, which may have a cardiovascular protective effect. However, the regulation and function of sEH in cancer are largely unknown. The present study investigated whether DNA methylation regulates the expression of sEH in carcinoma HepG2 cells. The mRNA and protein expressions of sEH in HepG2 cells were lower than those in transformed human embryonic kidney cells and in primary cultured human endothelial cells. Bioinformatic analysis revealed a putative CpG island and 5 SP-1 binding sites located in the promoter region of the sEH gene. Furthermore, the sEH expression was significantly enhanced by demethylation treatment with 5-Aza-CdR, a DNA methyltransferase inhibitor, and the sEH promoter was transformed from hypermethylation to hypomethylation as detected by methylation-specific PCR and bisulfite sequencing. Transient transfection assays showed that the activity of the human sEH promoter was increased in HepG2 cells in response to 5-Aza-CdR. Five SP-1 binding sites in the promoter region responding to treatment with 5-Aza-CdR were identified by construct deletion and mutation analysis and chromatin immunoprecipitation assay. Interestingly, adenoviral overexpression of sEH in HepG2 cells decreased cell proliferation. Thus, SP-1 is involved in the decrease in the transcription of sEH as a result of DNA methylation in HepG2 cells, which might contribute to epigenetic mechanism-induced carcinogenesis in hepatocytes.
ESTHER : Zhang_2010_Biochim.Biophys.Acta_1799_659
PubMedSearch : Zhang_2010_Biochim.Biophys.Acta_1799_659
PubMedID: 20888937

Title : Patients achieving clearance of HCV with interferon therapy recover from decreased retinol-binding protein 4 levels - Iwasa_2009_J.Viral.Hepat_16_716
Author(s) : Iwasa M , Hara N , Miyachi H , Tanaka H , Takeo M , Fujita N , Kobayashi Y , Kojima Y , Kaito M , Takei Y
Ref : J Viral Hepat , 16 :716 , 2009
Abstract : Retinol-binding protein 4 (RBP4) is a recently identified adipokine that is elevated in the blood in several insulin-resistant states. We investigated the association between plasma RBP4 and histological and biochemical characteristics of chronic hepatitis C (CHC), as well as changes in RBP4 levels following interferon therapy. Eighty-one patients with CHC infected with genotype 1 received treatment with peginterferon plus ribavirin. Histological data were available for 41 out of 81 patients before treatment, and the degree of fibrosis, inflammation and steatosis was assessed. Plasma levels of RBP4 were determined in serial samples (before, at the end of treatment, and at 6 months post-treatment). RBP4 levels were lower in CHC patients than in control subjects (34.6 +/- 12.3 microg/mL vs 46.2 +/- 10.5 microg/mL; P
ESTHER : Iwasa_2009_J.Viral.Hepat_16_716
PubMedSearch : Iwasa_2009_J.Viral.Hepat_16_716
PubMedID: 19302338

Title : Role of endothelial lipase in plasma HDL levels in a murine model of hypertriglyceridemia - Tanaka_2009_J.Atheroscler.Thromb_16_327
Author(s) : Tanaka H , Ishida T , Johnston TP , Yasuda T , Ueyama T , Kojima Y , Kundu RK , Quertermous T , Ishikawa Y , Hirata K
Ref : J Atheroscler Thromb , 16 :327 , 2009
Abstract : AIM: Hypertriglyceridemia is the most common cause of low plasma high-density lipoprotein cholesterol (HDL-C) levels; however, the correlation between high triglyceride (TG) and low HDL-C remains unclear. Endothelial lipase (EL) is a determinant of plasma HDL levels. We investigated the role of EL in HDL metabolism in a murine model of acute hypertriglyceridemia. METHODS AND RESULTS: To establish TG-dominant hyperlipidemia, EL-/- and wild-type (WT) mice were injected with Poloxamer-407 (P-407, 0.5 g/kg, i.p.). A single injection of P-407 resulted in a marked increase in plasma TG and cholesterol levels together with a decrease in HDL-C levels. Although plasma TG levels were similar in EL-/- and WT mice after P-407 injection, HDL-C levels were 80% higher and the HDL particle size was significantly larger in EL-/- mice than in WT mice. P-407 treatment inhibited plasma lipoprotein lipase activity and EL phospholipase activity, without decreasing their expressions. Adenovirus-mediated overexpression of EL in the liver reduced plasma HDL-C levels in both normo- and hyperlipidemic mice, while overexpression of catalytically inactive EL reduced HDL-C levels in hyperlipidemic mice. Cell culture experiments revealed that both catalytically active and inactive EL promoted cellular HDL uptake to the same extent. CONCLUSION: EL regulates plasma HDL levels in mice in the normolipidemic as well as the acute hypertriglyceridemic state. EL can modulate plasma HDL-CHOL levels through both its lipolytic and ligand-binding functions in hypertriglyceridemic mice, while lipolytic activity appears to be the main determinant for its effects on HDL metabolism in normolipidemic mice.
ESTHER : Tanaka_2009_J.Atheroscler.Thromb_16_327
PubMedSearch : Tanaka_2009_J.Atheroscler.Thromb_16_327
PubMedID: 19672025
Gene_locus related to this paper: human-LIPG

Title : The effects of fasting and general anesthesia on serum chemistries in KCG miniature pigs - Tanaka_2009_J.Am.Assoc.Lab.Anim.Sci_48_33
Author(s) : Tanaka H , Igarashi T , Lefor AT , Kobayashi E
Ref : J Am Assoc Lab Anim Sci , 48 :33 , 2009
Abstract : Investigators are obligated to optimize the perioperative care of experimental animals, but little is known about the effects of anesthesia and surgery on serum chemistries in KCG pigs. The objective of this study was to examine the influence of fasting and surgery under general anesthesia on 27 serum chemistries in KCG miniature pigs to improve management. Crossbred KCG minipigs were used at a mean of 12.3 mo of age (range, 8.6 to 14.9) and 33.4 kg of body weight (range, 24.0 to 40.2). Serum chemistries were evaluated at the start and end of a 24 h fasting period in fasted animals (n = 6). No significant differences were observed between the starting and postfasting studies. Partial hemilaminectomy of the lumbar spine was carried out in 2 groups of animals. Those given sevoflurane anesthesia (n = 7) had significant decreases in serum albumin, potassium, inorganic phosphorus, gamma-glutamyltransferase peptidase, cholinesterase, and glucose postoperatively compared with preoperative values. Animals given isoflurane (n = 7) anesthesia had significantly decreased total protein, albumin, triglyceride, phospholipids, sodium, potassium, calcium, alanine aminotransferase, alkaline phoshatase and glucose after surgery compared with levels before surgery. In a separate experiment (n = 7), serum glucose and insulin also decreased during the postoperative period after isoflurane anesthesia. These results demonstrate that select serum electrolytes, glucose, and insulin of KCG miniature pigs are altered after general anesthesia. Investigators must be aware of the effects of anesthetic agents on experimental animals to provide optimal care and for interpretation of experimental data.
ESTHER : Tanaka_2009_J.Am.Assoc.Lab.Anim.Sci_48_33
PubMedSearch : Tanaka_2009_J.Am.Assoc.Lab.Anim.Sci_48_33
PubMedID: 19245748

Title : Inhibition of acetylcholinesterase by metabolites of copper pyrithione (CuPT) and its possible involvement in vertebral deformity of a CuPT-exposed marine teleostean fish - Mochida_2009_Comp.Biochem.Physiol.C.Toxicol.Pharmacol_149_624
Author(s) : Mochida K , Ito K , Harino H , Tanaka H , Onduka T , Kakuno A , Fujii K
Ref : Comparative Biochemistry & Physiology C Toxicol Pharmacol , 149 :624 , 2009
Abstract : In a previous study, we demonstrated that exposure to an antifouling biocide, copper pyrithione (CuPT), early during life induced vertebral deformity in the larvae of a marine fish, the mummichog (Fundulus heteroclitus). Skeletal deformities may be caused by inhibition by of acetylcholiensterase (AChE) activity, and to elucidate the mechanism underlying the CuPT-associated vertebral deformity, we first examined whether CuPT, zinc pyrithione (ZnPT), and their degradation products could inhibit AChE activity in the fish. Two of the degradation products, 2,2'-dipyridyldisulfide [(PS)(2)] and 2,2'-dithiobispyridine-N-oxide [(PT)(2)], but neither CuPT nor ZnPT, exhibited prominent AChE-inhibiting activity. Secondly, thin-layer chromatography revealed that mummichog hepatic microsomes metabolized CuPT to produce (PS)(2) in a microsome-dependent manner. The AChE inhibition induced in CuPT-exposed fish is likely due to (PS)(2) that was produced through metabolism of acquired CuPT. (PS)(2) may cause therefore skeletal deformity in CuPT-exposed fish by means of its neuromuscular blocking properties, through a mechanism similar to that proposed for animals exposed to organophosphorous pesticides.
ESTHER : Mochida_2009_Comp.Biochem.Physiol.C.Toxicol.Pharmacol_149_624
PubMedSearch : Mochida_2009_Comp.Biochem.Physiol.C.Toxicol.Pharmacol_149_624
PubMedID: 19211040

Title : The genome of a lepidopteran model insect, the silkworm Bombyx mori - Xia_2008_Insect.Biochem.Mol.Biol_38_1036
Author(s) : Xia Q , Wang J , Zhou Z , Li R , Fan W , Cheng D , Cheng T , Qin J , Duana J , Xu H , Li Q , Li N , Wang M , Dai F , Liu C , Lin Y , Zhao P , Zhang H , Liu S , Zha X , Li C , Zhao A , Pan M , Pan G , Shen Y , Gao Z , Wang Z , Wang G , Wu Z , Hou Y , Chai C , Yu Q , He N , Zhang Z , Li S , Yang H , Lu C , Xiang Z , Mita K , Kasahara M , Nakatani Y , Yamamoto K , Abe H , Ahsan B , Daimoni T , Doi K , Fujii T , Fujiwara H , Fujiyama A , Futahashi R , Hashimotol S , Ishibashi J , Iwami M , Kadono-Okuda K , Kanamori H , Kataoka H , Katsuma S , Kawaoka S , Kawasaki H , Kohara Y , Kozaki T , Kuroshu RM , Kuwazaki S , Matsushima K , Minami H , Nagayasu Y , Nakagawa T , Narukawa J , Nohata J , Ohishi K , Ono Y , Osanai-Futahashi M , Ozaki K , Qu W , Roller L , Sasaki S , Sasaki T , Seino A , Shimomura M , Shin-I T , Shinoda T , Shiotsuki T , Suetsugu Y , Sugano S , Suwa M , Suzuki Y , Takiya S , Tamura T , Tanaka H , Tanaka Y , Touhara K , Yamada T , Yamakawa M , Yamanaka N , Yoshikawa H , Zhong YS , Shimada T , Morishita S
Ref : Insect Biochemistry & Molecular Biology , 38 :1036 , 2008
Abstract : Bombyx mori, the domesticated silkworm, is a major insect model for research, and the first lepidopteran for which draft genome sequences became available in 2004. Two independent data sets from whole-genome shotgun sequencing were merged and assembled together with newly obtained fosmid- and BAC-end sequences. The remarkably improved new assembly is presented here. The 8.5-fold sequence coverage of an estimated 432 Mb genome was assembled into scaffolds with an N50 size of approximately 3.7 Mb; the largest scaffold was 14.5 million base pairs. With help of a high-density SNP linkage map, we anchored 87% of the scaffold sequences to all 28 chromosomes. A particular feature was the high repetitive sequence content estimated to be 43.6% and that consisted mainly of transposable elements. We predicted 14,623 gene models based on a GLEAN-based algorithm, a more accurate prediction than the previous gene models for this species. Over three thousand silkworm genes have no homologs in other insect or vertebrate genomes. Some insights into gene evolution and into characteristic biological processes are presented here and in other papers in this issue. The massive silk production correlates with the existence of specific tRNA clusters, and of several sericin genes assembled in a cluster. The silkworm's adaptation to feeding on mulberry leaves, which contain toxic alkaloids, is likely linked to the presence of new-type sucrase genes, apparently acquired from bacteria. The silkworm genome also revealed the cascade of genes involved in the juvenile hormone biosynthesis pathway, and a large number of cuticular protein genes.
ESTHER : Xia_2008_Insect.Biochem.Mol.Biol_38_1036
PubMedSearch : Xia_2008_Insect.Biochem.Mol.Biol_38_1036
PubMedID: 19121390
Gene_locus related to this paper: bommo-a0mnw6 , bommo-a1yw85 , bommo-a9ls22 , bommo-ACHE1 , bommo-ACHE2 , bommo-b0fgv8 , bommo-b1q137 , bommo-b1q139 , bommo-b1q140 , bommo-b1q141 , bommo-b2zdz0 , bommo-b3gef6 , bommo-b3gef7 , bommo-b3gs55 , bommo-b3gs56 , bommo-d2ktu3 , bommo-d2ktu5 , bommo-d9ile0 , bommo-e1cga5 , bommo-e1cga6 , bommo-g8fpz6 , bommo-h9iu43 , bommo-h9iu46 , bommo-h9iu47.1 , bommo-h9iu47.2 , bommo-h9iue5 , bommo-h9ivg2 , bommo-h9iwj7 , bommo-h9iwj8 , bommo-h9ix58 , bommo-h9ixi1.1 , bommo-h9ixi1.2 , bommo-h9iy47 , bommo-h9izw1 , bommo-h9j0s4 , bommo-h9j1y0 , bommo-h9j3r0 , bommo-h9j3w6 , bommo-h9j3w7 , bommo-h9j5t0 , bommo-h9j8g3 , bommo-h9j9k9 , bommo-h9j066 , bommo-h9j067 , bommo-h9j593 , bommo-h9j594 , bommo-h9j990 , bommo-h9jde8 , bommo-h9jde9 , bommo-h9jdf0 , bommo-h9jds4 , bommo-h9jle7 , bommo-h9jn83 , bommo-h9jn85 , bommo-h9jrg2 , bommo-h9jyh9 , bommo-JHE , bommo-m1rmh6 , bommo-q1hq05 , bommo-q4tte1 , bommo-h9j592 , bommo-h9j604 , bommo-h9jpm8 , bommo-h9iss4 , bommo-h9j2c7

Title : Chinese dumpling scare hits Japan--a case of methamidophos food poisoning - Sumi_2008_J.Toxicol.Sci_33_485
Author(s) : Sumi Y , Oode Y , Tanaka H
Ref : Journal of Toxicological Sciences , 33 :485 , 2008
Abstract : An outbreak of food poisoning that affected at least ten people in various regions of Japan was traced to exposure to Chinese dumplings contaminated with the organophosphate insecticide Methamidophos. We experienced the most serious case, a five years old girl, who suffered coma. She presented with features of cholinergic overactivity and her serum cholinesterase activity was 9 U/l. We started intravenous treatment with pralidoxime iodide, atropine sulfate, and midazolam. Her symptoms improved gradually and she was discharged on day 25 without any sequelae. Though poisoning attributed to organophosphate insecticides has become less common in recent years, it is even more important to diagnose the problem rapidly based on the characteristic symptoms and to start specific treatment at the earliest possible stage after poisoning.
ESTHER : Sumi_2008_J.Toxicol.Sci_33_485
PubMedSearch : Sumi_2008_J.Toxicol.Sci_33_485
PubMedID: 18827448

Title : Transcriptional regulation of the human soluble epoxide hydrolase gene EPHX2 - Tanaka_2008_Biochim.Biophys.Acta_1779_17
Author(s) : Tanaka H , Kamita SG , Wolf NM , Harris TR , Wu Z , Morisseau C , Hammock BD
Ref : Biochimica & Biophysica Acta , 1779 :17 , 2008
Abstract : Soluble epoxide hydrolase (sEH) is a multifunctional protein encoded by the EPHX2 gene. The biological functions and enzyme kinetics of sEH have been extensively investigated, however, little is known about its transcriptional regulation and mechanisms of tissue specific expression. Here, a luciferase gene based reporter assay was used to identify the minimal promoter and cis regulatory elements of EPHX2. The minimal promoter was identified as a GC-rich region between nts -374 and +28 with respect to the putative transcriptional start site. A reporter plasmid carrying this minimal promoter showed higher or similar activities in comparison to a reporter plasmid carrying nts -5,974 to +28 of EPHX2 in 9 human cell lines that were tested. Sp1 binding sites that are involved in augmenting the minimal promoter activity of EPHX2 were identified by nested deletion analysis, site-specific mutation, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay.
ESTHER : Tanaka_2008_Biochim.Biophys.Acta_1779_17
PubMedSearch : Tanaka_2008_Biochim.Biophys.Acta_1779_17
PubMedID: 18078836

Title : Identification of two epoxide hydrolases in Caenorhabditis elegans that metabolize mammalian lipid signaling molecules - Harris_2008_Arch.Biochem.Biophys_472_139
Author(s) : Harris TR , Aronov PA , Jones PD , Tanaka H , Arand M , Hammock BD
Ref : Archives of Biochemistry & Biophysics , 472 :139 , 2008
Abstract : We have identified two genes in the genomic database for Caenorhabditis elegans that code for proteins with significant sequence similarity to the mammalian soluble epoxide hydrolase (sEH). The respective transcripts were cloned from a mixed stage cDNA library from C. elegans. The corresponding proteins obtained after recombinant expression in insect cells hydrolyzed standard epoxide hydrolase substrates, including epoxyeicosatrienoic acids (EETs) and leukotoxins (EpOMEs). The enzyme activity was inhibited by urea-based compounds originally designed to inhibit the mammalian sEH. In vivo inhibition of the enzymes using the most potent of these compounds resulted in elevated levels of the EpOMEs in the nematode. These results suggest that the hydrolases are involved in the metabolism of possible lipid signaling molecules in C. elegans.
ESTHER : Harris_2008_Arch.Biochem.Biophys_472_139
PubMedSearch : Harris_2008_Arch.Biochem.Biophys_472_139
PubMedID: 18267101
Gene_locus related to this paper: caeel-K07C5.5

Title : Endothelial lipase is increased by inflammation and promotes LDL uptake in macrophages - Yasuda_2007_J.Atheroscler.Thromb_14_192
Author(s) : Yasuda T , Hirata K , Ishida T , Kojima Y , Tanaka H , Okada T , Quertermous T , Yokoyama M
Ref : J Atheroscler Thromb , 14 :192 , 2007
Abstract : AIM: Endothelial lipase (EL) is a member of the lipoprotein lipase family that regulates HDL metabolism. EL is known to act as a bridging molecule for monocytes or lipoproteins in vascular endothelial cells. We investigated the role and regulatory mechanisms of EL expression in macrophages.
METHODS: Macrophages originating from wild-type (EL+/+) and EL-deficient (EL-/-) mice were stimulated with lipopolysaccharide (LPS). The expression of EL mRNA was evaluated by northern blotting. DiI-LDL was used to measure the uptake of native low-density lipoprotein (nLDL).
RESULTS: LPS increased EL mRNA levels by increasing intracellular oxidative stress in the macrophages. LPS did not affect EL expression in macrophages derived from Toll-like receptor 4 (TLR4) gene mutant mice, C3H/HeJ. The uptake of nLDL after LPS-treatment was significantly lower in macrophages from EL-/- mice than those from EL+/+ mice. Simvastatin suppressed the LPS-induced upregulation of EL expression and uptake of nLDL.
CONCLUSIONS: EL expression is upregulated by LPS via TLR4 and promotes the uptake of nLDL by macrophages. Simvastatin inhibits the LPS-induced up-regulation and uptake in macrophages. Thus, our findings provide a novel role for EL in lipoprotein metabolism and would expand the range of anti-atherogenic effects of statins.
ESTHER : Yasuda_2007_J.Atheroscler.Thromb_14_192
PubMedSearch : Yasuda_2007_J.Atheroscler.Thromb_14_192
PubMedID: 17726294

Title : Analysis of two acetylcholinesterase genes in Bombyx mori - Seino_2007_Pestic.Biochem.Physiol_88_92
Author(s) : Seino A , Kazuma T , Tan AJ , Tanaka H , Kono Y , Mita K , Shiotsuki T
Ref : Pesticide Biochemistry and Physiology , 88 :92 , 2007
Abstract : Two previous acetylcholinesterases (AChE, EC 3.1.1.7) cDNAs were identified and cloned from silkworm, Bombyx mori. One of those, BmAChE-o cDNA, is comprised of 3197 nucleotides which encode 638 amino acids, having an amino acid sequence homology of 72% with Drosophila melanogaster Ace-orthologous AChE (AO-AChE). In some species, another AChE group based on the sequence, Drosophila Ace-paralogous AChE (AP-AChE) has been recognized in relation to organophosphate- or carbamate-resistance, but there have been few reports of AP-AChE among lepidopteran species. However, we isolated the AP-AChE from lepidopteran silkworm, and cloned full ORF as BmAChE-p, which cDNA consisted of 2465 nucleotides that encode 683 amino acids. The homologies with other AP-AChEs were over 60% when compared. Although silkworm is not a target of pesticides, the genomic information obtained in this study will contribute to insecticide-resistance study on lepidopteran pest species.
ESTHER : Seino_2007_Pestic.Biochem.Physiol_88_92
PubMedSearch : Seino_2007_Pestic.Biochem.Physiol_88_92
PubMedID:
Gene_locus related to this paper: bommo-ACHE1 , bommo-ACHE2

Title : Femoral neck fracture as a complication of lipase-secreting pancreatic acinar cell carcinoma -
Author(s) : Hashimoto M , Miki K , Beck Y , Kokudo N , Makuuchi M , Tanaka H
Ref : Surgery , 142 :779 , 2007
PubMedID: 17981202

Title : CD26 mediates dissociation of Tollip and IRAK-1 from caveolin-1 and induces upregulation of CD86 on antigen-presenting cells - Ohnuma_2005_Mol.Cell.Biol_25_7743
Author(s) : Ohnuma K , Yamochi T , Uchiyama M , Nishibashi K , Iwata S , Hosono O , Kawasaki H , Tanaka H , Dang NH , Morimoto C
Ref : Molecular & Cellular Biology , 25 :7743 , 2005
Abstract : CD26 is a T-cell costimulatory molecule with dipeptidyl peptidase IV enzyme activity in its extracellular region. We have previously reported that the addition of recombinant soluble CD26 resulted in enhanced proliferation of human T lymphocytes induced by the recall antigen tetanus toxoid (TT) via upregulation of CD86 on monocytes and that caveolin-1 was a binding protein of CD26, and the CD26-caveolin-1 interaction resulted in caveolin-1 phosphorylation (p-cav-1) as well as TT-mediated T-cell proliferation. However, the mechanism involved in this immune enhancement has not yet been elucidated. In the present work, we perform experiments to identify the molecular mechanisms by which p-cav-1 leads directly to the upregulation of CD86. Through proteomic analysis, we identify Tollip (Toll-interacting protein) and IRAK-1 (interleukin-1 receptor-associated serine/threonine kinase 1) as caveolin-1-interacting proteins in monocytes. We also demonstrate that following stimulation by exogenous CD26, Tollip and IRAK-1 dissociate from caveolin-1, and IRAK-1 is then phosphorylated in the cytosol, leading to the upregulation of CD86 via activation of NF-kappaB. Binding of CD26 to caveolin-1 therefore regulates signaling pathways in antigen-presenting cells to induce antigen-specific T-cell proliferation.
ESTHER : Ohnuma_2005_Mol.Cell.Biol_25_7743
PubMedSearch : Ohnuma_2005_Mol.Cell.Biol_25_7743
PubMedID: 16107720

Title : Precocious metamorphosis in transgenic silkworms overexpressing juvenile hormone esterase - Tan_2005_Proc.Natl.Acad.Sci.U.S.A_102_11751
Author(s) : Tan A , Tanaka H , Tamura T , Shiotsuki T
Ref : Proc Natl Acad Sci U S A , 102 :11751 , 2005
Abstract : Insect growth and development are intricately regulated by the titers of juvenile hormones (JHs) and ecdysteroids (and/or their metabolites) in the insect hemolymph. Hydrolysis of the methyl ester of JH by a JH-specific esterase (JHE) is a key pathway for the degradation of JH. Here, we generate transgenic silkworm strains that overexpress JHE by using the binary GAL4/UAS system. Overexpression of JHE from the embryonic stage resulted in larval-pupal metamorphosis after the third stadium, two stadia earlier than that observed in wild-type insects. This precocious metamorphosis suggests that JHs are not critical for normal development of embryo or larva before the second molt in Lepidoptera (moths and butterflies). Our transgenic approach allowed us to dissect the function of key physiological events that occur from embryogenesis. Until now, these types of studies were possible only in later larval stadia by using physical techniques such as allatectomy or the application of JH analogues. We believe that our system will allow further pioneering studies in insect physiology.
ESTHER : Tan_2005_Proc.Natl.Acad.Sci.U.S.A_102_11751
PubMedSearch : Tan_2005_Proc.Natl.Acad.Sci.U.S.A_102_11751
PubMedID: 16087883

Title : A lipase isolated from the silkworm Bombyx mori shows antiviral activity against nucleopolyhedrovirus - Ponnuvel_2003_J.Virol_77_10725
Author(s) : Ponnuvel KM , Nakazawa H , Furukawa S , Asaoka A , Ishibashi J , Tanaka H , Yamakawa M
Ref : J Virol , 77 :10725 , 2003
Abstract : A protein showing strong antiviral activity against Bombyx mori nucleopolyhedrovirus (BmNPV) was purified from the digestive juice of B. mori larvae. A homology search of the deduced amino acid sequence of the protein cDNA revealed 56% homology with Drosophila melanogaster lipase and 21% homology with human lipase. As lipase activity of the protein was confirmed in vitro, this protein was designated Bmlipase-1. Northern blot analysis showed that the Bmlipase-1 gene is expressed in the midgut but not in other tissues, nor is it activated by BmNPV infection. In addition, the Bmlipase-1 gene was shown not to be expressed in the molting and wandering stages, indicating that the gene is hormonally regulated. Our results suggest that an insect digestive enzyme has potential as a physiological barrier against BmNPV at the initial site of viral infection.
ESTHER : Ponnuvel_2003_J.Virol_77_10725
PubMedSearch : Ponnuvel_2003_J.Virol_77_10725
PubMedID: 12970462
Gene_locus related to this paper: bommo-Q8MY66

Title : Expression of pancreatic enzyme genes during the early larval stage of Japanese eel, Anguilla japonica. -
Author(s) : Kurokawa T , Suzuki T , Ohta H , Kagawa H , Tanaka H , Unuma T
Ref : Fish Sci , 68 :736 , 2002
PubMedID:
Gene_locus related to this paper: angja-1plip

Title : Antitumor efficacy of hypothemycin, a new Ras-signaling inhibitor - Tanaka_1999_Jpn.J.Cancer.Res_90_1139
Author(s) : Tanaka H , Nishida K , Sugita K , Yoshioka T
Ref : Jpn J Cancer Research , 90 :1139 , 1999
Abstract : We have devised a new drug screening assay to discover anti-cancer drugs which inhibit Ras-mediated cellular signals, by utilizing a Ras-responsive element (RRE)-driven reporter gene system. We found that hypothemycin, an anti-bacterial, reduces RRE-dependent transcription. Treatment of tumor cells with hypothemycin resulted in reduced expression of Ras-inducible genes, including MMP (matrix metalloproteinase)-1, MMP-9, transforming growth factor-beta (TGF-beta), and vascular endothelial growth factor (VEGF), but not that of the constitutively expressed gene, MMP-2. The results of zymography demonstrated that hypothemycin reduced the production of MMP-9 and MMP-3, another Ras-inducible MMP, in the culture medium. Hypothemycin selectively inhibits anchorage-independent growth of Ras-transformed cells in comparison with anchorage-dependent growth. These findings suggest that hypothemycin inhibits Ras-mediated cellular signaling. Daily treatment of tumor-bearing mice with hypothemycin resulted in significant inhibition of tumor growth. Since MMP-1, MMP-3 and MMP-9 play important roles in tumor invasion and TGF-beta and VEGF are involved in tumor angiogenesis, hypothemycin is considered to be an example of a new class of antitumor drugs, whose antitumor efficacy can be at least partly attributed to inhibition of Ras-inducible genes.
ESTHER : Tanaka_1999_Jpn.J.Cancer.Res_90_1139
PubMedSearch : Tanaka_1999_Jpn.J.Cancer.Res_90_1139
PubMedID: 10595743

Title : Acetylcholinesterase activity, and neurofilament protein, and catecholamine synthesizing enzymes immunoreactivities in the mouse adrenal gland during postnatal development - Iwasa_1999_J.Vet.Med.Sci_61_621
Author(s) : Iwasa K , Oomori Y , Tanaka H
Ref : J Vet Med Sci , 61 :621 , 1999
Abstract : The present study showed the acetylcholinesterase (AChE) activity, and neurofilament protein (NFP), catecholamine-synthesizing enzymes, dopamine beta-hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT) immunoreactivities in the mouse adrenal gland during postnatal development. From birth to postnatal-1-day, AChE activity was weakly and diffusely found in some medullary cells and in very few nerve fibers whereas strong NFP immunoreactivity was seen in a few ganglion cells and in remarkably numerous nerve fibers in the medulla. Almost all meduallary cells were reactive for both DBH and PNMT during this period. From postnatal-2- or -3-day to postnatal-1-week, strong AChE activity was observed in a few large ganglion cells, but the reaction was weak in clusters of chromaffin cells, and the number of strong AChE-active nerve fibers in the medulla was rapidly increased. From postnatal-2-day onwards, the number of NFP-immunoreactive nerve fibers in the medulla were remarkably numerous. Numerous chromaffin cells were reactive for both DBH and PNMT whereas some chromaffin cells were reactive for only DBH from postnatal-2-day onwards. These results suggest that drastic changes such as an increase of acetylcholine in the nerve fibers, differentiation of noradrenaline and adrenaline cells of the medulla may occur during this period. From postnatal-2-week to postnatal-3-week, weak AChE activity was seen in the clusters of several chromaffin cells and a few ganglion cells, and the number of AChE-active nerve fibers in the medulla was gradually increased. From postnatal-4-week to postnatal-8-week (adult), the distribution and frequency of AChE activity in the adrenal gland were similar to those at postnatal-3-week. In the adult, AChE activity was weakly seen in the clusters of several chromaffin cells showing noradrenaline fluorescence in the adrenal medulla. The noradrenaline cells were contacted by denser AChE-reactive nerve fibers than adrenaline cells. These results suggest that the development of cholinergic nervous system in the mouse adrenal medulla may be completed by postnatal-3-week.
ESTHER : Iwasa_1999_J.Vet.Med.Sci_61_621
PubMedSearch : Iwasa_1999_J.Vet.Med.Sci_61_621
PubMedID: 10423683

Title : Colocalization of gamma-aminobutyric acid immunoreactivity and acetylcholinesterase activity in nerve fibers of the mouse adrenal gland - Iwasa_1999_J.Vet.Med.Sci_61_631
Author(s) : Iwasa K , Oomori Y , Tanaka H
Ref : J Vet Med Sci , 61 :631 , 1999
Abstract : The present immunohistochemical and enzyme histochemical study showed gamma-aminobutyric acid (GABA) immunoreactivity and acetylcholinesterase (AChE) activity in the mouse adrenal gland. Weak GABA immunoreactivity was seen in clusters of chromaffin cells showing noradrenaline fluorescence. This finding suggests that both GABA and noradrenaline may be released from the granules of noradrenaline cells by adequate stimuli. GABA-immunoreactive varicose nerve fibers densely contacted adrenaline cells and large ganglion cells, but they were sparse in the periphery of clusters of noradrenaline cells. AChE activity was strong in a few large ganglion cells and weak in chromaffin cells showing noradrenaline fluorescence, and was found in numerous nerve bundles and fibers of the medulla. AChE-active nerve fibers more densely contacted noradrenaline cells than adrenaline cells. By using double labeling technique, numerous GABA-immunoreactive nerve fibers in the medulla were reactive for AChE in the same sections. These results suggest that both GABA and acetylcholine may be colocalized in the intra-adrenal nerve fibers and may have some secretory effects on the chromaffin cells.
ESTHER : Iwasa_1999_J.Vet.Med.Sci_61_631
PubMedSearch : Iwasa_1999_J.Vet.Med.Sci_61_631
PubMedID: 10423684

Title : Neuronal differentiation of neuro 2a cells by inhibitors of cell cycle progression, trichostatin A and butyrolactone I - Inokoshi_1999_Biochem.Biophys.Res.Commun_256_372
Author(s) : Inokoshi J , Katagiri M , Arima S , Tanaka H , Hayashi M , Kim YB , Furumai R , Yoshida M , Horinouchi S , Omura S
Ref : Biochemical & Biophysical Research Communications , 256 :372 , 1999
Abstract : Trichostatin A (TSA, 17 nM), a specific and reversible inhibitor of histone deacetylase induced neurite network formation at and after 4 days. The networks were preserved for at least 3 weeks in the presence of TSA. Butyrolactone I (BLI, 23.6 microM), an inhibitor of cdc2 and cdk2 kinases, also induced neurite extension. Both compounds enhanced the acetylcholinesterase activity of the cells. Cell cycle progression of the cells was blocked by TSA (17 nM) at G1 phase alone. Furthermore, the level of histone hyperacetylation and p21(WAF1) expression in TSA-treated cells increased transiently. These findings suggest that the induction of the neuronal differentiation in Neuro 2a cells by these agents requires the cell cycle arrest at G1 phase, which is caused by inhibition of cycline dependent kinase, a target molecule of BLI and p21(WAF1).
ESTHER : Inokoshi_1999_Biochem.Biophys.Res.Commun_256_372
PubMedSearch : Inokoshi_1999_Biochem.Biophys.Res.Commun_256_372
PubMedID: 10079191

Title : Purification and characterization of bovine pancreatic bile salt-activated lipase - Tanaka_1999_J.Biochem_125_883
Author(s) : Tanaka H , Mierau I , Ito F
Ref : J Biochem , 125 :883 , 1999
Abstract : An enzyme with lipase and esterase activity was purified from bovine pancreas. Furthermore, a non-radioactive lipase assay was developed which is 100 times more sensitive than the conventional methods and allowed the characterization of the lipase activity of the enzyme. The lipase activity increased 42 times in the presence of 10 mM sodium taurocholate, which for the first time provides direct evidence that a bile salt-activated lipase (bp-BAL) was isolated from bovine pancreas. This conclusion is further supported by the fact that the N-terminal amino acid sequence of this lipase/esterase is 88% homologous to human milk BAL and human pancreatic BAL. Staining with various lectins showed that bp-BAL is a glycoprotein which contains fucose residues. Previously from bovine pancreas a lysophospholipase has been purified and a gene was cloned and sequenced encoding an enzyme with cholesterol esterase/lysophospholipase activity. Comparison of the N-terminal amino acid sequence of bp-BAL with the deduced amino acid sequence of the latter revealed that they are identical. Furthermore, the molecular weight of the purified bp-BAL of 63,000, as estimated by SDS-PAGE, is very similar to that of the purified lysophospholipase (65,000) and to the theoretical molecular weight of 65,147 of the cholesterol esterase/lysophospholipase. These data suggest that these three enzymes are one and the same.
ESTHER : Tanaka_1999_J.Biochem_125_883
PubMedSearch : Tanaka_1999_J.Biochem_125_883
PubMedID: 10220579
Gene_locus related to this paper: bovin-balip

Title : Neuronal differentiation of Neuro 2a cells by lactacystin and its partial inhibition by the protein phosphatase inhibitors calyculin A and okadaic acid - Tanaka_1995_Biochem.Biophys.Res.Commun_216_291
Author(s) : Tanaka H , Katagiri M , Arima S , Matsuzaki K , Inokoshi J , Omura S
Ref : Biochemical & Biophysical Research Communications , 216 :291 , 1995
Abstract : Lactacystin (1.3 microM), a metabolite from an actinomycete, induced the formation of bipolar projections at both sides of the cell body of Neuro 2a cells 1 day after treatment and networks at and after 3 days and enhanced acetylcholinesterase activity (a marker of neuronal differentiation). Thus, the neuronal differentiation was characterized both morphologically and functionally. The experiments with various inhibitors of protein kinases and phosphatases revealed that the protein phosphatase inhibitors calyculin A (0.5 nM) and okadaic acid (0.6 nM) inhibit the formation of bipolar projections at 1 day, but does not inhibit the network formation at and after 3 days.
ESTHER : Tanaka_1995_Biochem.Biophys.Res.Commun_216_291
PubMedSearch : Tanaka_1995_Biochem.Biophys.Res.Commun_216_291
PubMedID: 7488103

Title : Effects of activin A\/erythroid differentiation factor on erythroid and megakaryocytic differentiations of mouse erythroleukemia (Friend) cells: evidence for two distinct modes of cell response - Okafuji_1995_Exp.Hematol_23_210
Author(s) : Okafuji K , Kaku K , Seguchi M , Tanaka H , Azuno Y , Kaneko T
Ref : Experimental Hematology , 23 :210 , 1995
Abstract : To further characterize activin A/erythroid differentiation factor (EDF) action on hematopoietic cell differentiation, we examined the effects of activin A/EDF on megakaryocytic and erythroid differentiation by determining acetylcholinesterase (AchE) activity and hemoglobin production in the mouse erythroleukemia (MEL) cell line F55. Activin A/EDF induced AchE activity of F55 cells in a dose-dependent manner. Erythroid differentiation of F55 cells, which was characterized by an increase in dianisidine-positive cells, was also induced by activin A/EDF. The effect of activin A/EDF on hemoglobin synthesis appeared more slowly compared with the effect on AchE activity. Erythroid differentiation induced by activin A/EDF was affected by the initial cell density, but AchE activity was not. Sodium orthovanadate, a tyrosine phosphatase inhibitor, markedly inhibited activin A/EDF-induced erythroid differentiation but not activin A/EDF-induced AchE activity. Other erythroid differentiation inducers, sodium butyrate and butyrylcholine chloride, mildly increased AchE activity in F55 cells, but N,N'-hexamethylene-bis-acetamide (HMBA), dimethyl sulfoxide (DMSO), and genistein did not. Dexamethasone inhibited HMBA-induced erythroid differentiation but did not affect activin A/EDF or sodium butyrate action. These results suggest that F55 cells potentially can differentiate into cells of a megakaryocytic lineage in addition to an erythroid lineage, and that activin A/EDF further potentiates the cell differentiation of this cell line. In addition, our results suggest that the mode of activin A/EDF effects on megakaryocytic differentiation is distinct from that on erythroid differentiation.
ESTHER : Okafuji_1995_Exp.Hematol_23_210
PubMedSearch : Okafuji_1995_Exp.Hematol_23_210
PubMedID: 7875239

Title : Alpha 1-adrenoceptors in the conduction system of rat hearts - Saito_1994_Br.J.Pharmacol_111_465
Author(s) : Saito K , Suetsugu T , Oku Y , Kuroda A , Tanaka H
Ref : British Journal of Pharmacology , 111 :465 , 1994
Abstract : 1. We have characterized alpha 1-adrenoceptor in the conduction systems of the rat heart by quantitative autoradiography. 2. Consecutive 20 micron thick sections from a single rat heart containing the sinoatrial (SA) node and atrioventricular (AV) node were incubated with increasing concentrations of [3H]-prazosin with or without 10 microM phentolamine. After exposure to 3H-Ultrofilm, optical densities corresponding to the SA node and AV node were determined by computerized densitometry after comparison with 3H standards. 3. The SA node and AV node were stained heavily for cholinesterase and they contained a higher concentration of alpha 1-adrenoceptors than the adjacent myocardium without a significant change in the affinity. 4. These results support the hypothesis that alpha 1-adrenoceptors may play an important role not only in inotropism but also in chronotropism of rat hearts.
ESTHER : Saito_1994_Br.J.Pharmacol_111_465
PubMedSearch : Saito_1994_Br.J.Pharmacol_111_465
PubMedID: 8004391

Title : Post-natal decrease in chronotropic sensitivity to acetylcholine in rat heart - Tanaka_1994_Gen.Pharmacol_25_157
Author(s) : Tanaka H , Matsuda T , Kawada H , Shigenobu K
Ref : General Pharmacology , 25 :157 , 1994
Abstract : 1. The negative chronotropic effects of acetylcholine and carbachol on isolated rat right atria were examined at 0, 4, 8, and 16 weeks after birth. 2. Acetylcholine produced negative chronotropic responses at all ages and completely abolished spontaneous beating at its maximum effective concentration. 3. The sensitivity to acetylcholine, expressed in terms of ED50 values, was higher at 0 and 4 weeks than at 8 and 16 weeks, ED50 values (microM) at 0, 4, 8 and 16 weeks being 9.5 +/- 1.8 (n = 12), 13.2 +/- 3.4 (n = 11), 59.3 +/- 10.9 (n = 14) and 51.5 +/- 17.5 (n = 5), respectively. 4. Neostigmine produced a leftward shift of the concentration-response curve for acetylcholine both at 4 and 8 weeks after birth. The shift was larger at 8 weeks and no difference in sensitivity to acetylcholine was observed between the two ages in the presence of neostigmine. 5. Further, no developmental changes were observed in the sensitivity to carbachol, which is not hydrolyzed by cholinesterase. 6. We concluded that the chronotropic sensitivity to acetylcholine of rat atria decreases post-natally during the period between 4 and 8 weeks after birth due to increase in cholinesterase activity.
ESTHER : Tanaka_1994_Gen.Pharmacol_25_157
PubMedSearch : Tanaka_1994_Gen.Pharmacol_25_157
PubMedID: 8026702

Title : Levels of mRNA coding for motoneuron growth-promoting factors are increased in denervated muscle - Rassendren_1992_Proc.Natl.Acad.Sci.U.S.A_89_7194
Author(s) : Rassendren FA , Bloch-Gallego E , Tanaka H , Henderson CE
Ref : Proc Natl Acad Sci U S A , 89 :7194 , 1992
Abstract : Partial denervation of skeletal muscle induces sprouting of axons remaining within the muscle, possibly as a result of increased synthesis by denervated muscle fibers of motoneuron growth-promoting factors. Direct verification of this hypothesis has not been possible because the molecules responsible are not unambiguously characterized. We used Xenopus oocytes as a functional assay for mRNAs coding for secreted growth factors: preparations of mRNA from innervated and denervated neonatal muscle were injected into oocytes. Three days later, oocytes injected with denervated muscle mRNA expressed increased levels of nicotinic acetylcholine receptor and voltage-dependent sodium channels at their membrane. Proteins secreted by the same oocytes were tested for their effects on (i) neurite outgrowth from embryonic chicken ventral spinal cord neurons; (ii) survival in mixed culture of embryonic chicken motoneurons identified using the SC1 antibody; and (iii) survival of embryonic motoneurons purified by panning on SC1 antibody. In all three assays, media conditioned by oocytes injected with mRNA from denervated muscle contained significantly higher levels of biological activity than did those from oocytes injected with innervated muscle mRNA or water. mRNA was prepared from muscle at different times after denervation: a maximal increase was obtained already after 1 day, consistent with an involvement in sprouting. Synthesis of motoneuron growth-promoting factors is thus regulated by denervation in a parallel fashion to that of other key components of the neuromuscular junction.
ESTHER : Rassendren_1992_Proc.Natl.Acad.Sci.U.S.A_89_7194
PubMedSearch : Rassendren_1992_Proc.Natl.Acad.Sci.U.S.A_89_7194
PubMedID: 1379735

Title : Cell death of lumbosacral motoneurons in chick, quail, and chick-quail chimera embryos: a test of the quantitative matching hypothesis of neuronal cell death - Tanaka_1986_J.Neurosci_6_2889
Author(s) : Tanaka H , Landmesser LT
Ref : Journal of Neuroscience , 6 :2889 , 1986
Abstract : The quantitative matching hypothesis of neuronal cell death was tested for the chick hindlimb by determining the relationship between myotube number at the onset of motoneuron cell death and the number of motoneurons that survive in chicks, quail, and chick-quail chimeras. Hindlimb buds, which differ in size between the 2 species, were exchanged at stages 16 1/2-19, myosin ATPase-stained myotubes in selected thigh muscles were counted during the cell death period (stages 30-34), and lumbosacral motoneurons were counted following the cell death period (stage 38). No quail motoneurons were rescued when quail cords innervated chick limbs. When chick cords innervated quail limbs, the number of surviving motoneurons was significantly decreased but not to quail values. We consider that this occurred because chicks develop more slowly than quail, and we found that transplanted chick limbs were developmentally younger than the contralateral quail limb at the onset of motoneuron cell death and contained fewer myotubes. Similarly, transplanted quail limbs contained more myotubes at the onset of cell death than normal stage 30 quail limbs. An excellent correlation was obtained during normal development of both species between the number of myotube clusters at the onset of cell death and the number of surviving motoneurons. This correlation was also observed for chick-quail chimeras, and when the data points were plotted for control chick, control quail, chick host-quail limb, and quail host-chick limb, the correlation coefficient was 0.996. This strongly suggests that some parameter closely related to myotube number limits the number of motoneurons that will survive. A proposal consistent with our observations is that motoneuron survival is dependent on the uptake of a myotube-derived trophic factor that can only be taken up at synaptic sites and that the number of such sites is limited and directly related to myotube number. In conclusion, our observations strongly support a quantitative-matching component in the process of neuronal cell death. However, since we were unable to rescue any neurons, we cannot exclude the possibility that some proportion of neurons normally dies for reasons other than peripheral competition.
ESTHER : Tanaka_1986_J.Neurosci_6_2889
PubMedSearch : Tanaka_1986_J.Neurosci_6_2889
PubMedID: 3760941

Title : Interspecies selective motoneuron projection patterns in chick-quail chimeras - Tanaka_1986_J.Neurosci_6_2880
Author(s) : Tanaka H , Landmesser LT
Ref : Journal of Neuroscience , 6 :2880 , 1986
Abstract : During normal development chick motoneurons have been shown to project selectively to appropriate muscles by responding to a series of cues, both specific and nonspecific, within the limb. We tested the ability of motoneurons from another avian species, the Japanese quail, to respond to these cues by transplanting chick limb buds onto quail embryos and quail limb buds onto chick embryos between stages 17 1/2 and 19. Feulgen staining, which distinguishes chick from quail cells on the basis of nuclear chromatin, revealed that all limb tissue, including muscle, was of donor origin, indicating that the migration of somite-derived muscle precursor cells had been completed by the time of transplantation. Normal quail motoneuron pools for most muscles were located in the same relative positions as homologous chick pools. In chick-quail chimeras we found that the motoneuron pools of one species selectively innervated the homologous muscles in the limb of opposite species with considerable precision. This was determined by defining the segmental innervation pattern of the muscles electrophysiologically and by retrogradely labeling motoneuron pools with HRP. Selective innervation was confirmed by using the functional activation patterns of the motoneuron pools as an additional means of identifying motoneurons. We conclude that any limb-derived cues required by motoneurons to project to their appropriate muscles must be similar in chick and quail and that the growth cones of both species must have similar detector systems for responding to these cues. Only 7 spinal segments were found to innervate the quail limb (versus 8 for the chick), resulting in an anterior shift in the spinal segments innervating several posterior quail muscles.(ABSTRACT TRUNCATED AT 250 WORDS)
ESTHER : Tanaka_1986_J.Neurosci_6_2880
PubMedSearch : Tanaka_1986_J.Neurosci_6_2880
PubMedID: 3020186