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  1. Article ; Online: Visualization of Reelin Secretion from Primary Cultured Neurons by Bioluminescence Imaging.

    Nakao, Yousuke / Yokawa, Satoru / Kohno, Takao / Suzuki, Takahiro / Hattori, Mitsuharu

    Journal of biochemistry

    2022  Volume 171, Issue 5, Page(s) 591–598

    Abstract: Reelin is a secreted glycoprotein important for brain development and synaptic plasticity in the adult brain. Some reports suggest that Reelin is secreted from the nerve terminals and functions as a neurotransmitter. However, the mechanism of Reelin ... ...

    Abstract Reelin is a secreted glycoprotein important for brain development and synaptic plasticity in the adult brain. Some reports suggest that Reelin is secreted from the nerve terminals and functions as a neurotransmitter. However, the mechanism of Reelin secretion is unknown. In this study, we visualized Reelin secretion by bioluminescence imaging using a fusion protein of Reelin and Gaussia luciferase (GLase-Reelin). GLase-Reelin expressed in HEK293T cells was correctly processed and secreted. Luminescence signals from the secreted GLase-Reelin of primary cultured neurons were visualized by bioluminescence microscopy. Reelin secretory events were observed at neurites and cell bodies. Bioluminescence imaging was also performed before and after KCl depolarization to compare the secretory events of Reelin and brain-derived neurotrophic factor (BDNF). The secretion of BDNF increased markedly shortly after depolarization. In contrast, the frequency of Reelin secretion did not change significantly by depolarization. Thus, Reelin secretion from neurites might not be regulated in a neuronal activity-dependent manner.
    MeSH term(s) Brain/metabolism ; Brain-Derived Neurotrophic Factor/metabolism ; Cell Adhesion Molecules, Neuronal/genetics ; Cell Adhesion Molecules, Neuronal/metabolism ; Cells, Cultured ; Extracellular Matrix Proteins/metabolism ; HEK293 Cells ; Humans ; Neurites/metabolism ; Neurons/metabolism
    Chemical Substances Brain-Derived Neurotrophic Factor ; Cell Adhesion Molecules, Neuronal ; Extracellular Matrix Proteins
    Language English
    Publishing date 2022-05-18
    Publishing country England
    Document type Journal Article
    ZDB-ID 218073-x
    ISSN 1756-2651 ; 0021-924X
    ISSN (online) 1756-2651
    ISSN 0021-924X
    DOI 10.1093/jb/mvac019
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  2. Article ; Online: Involvement of intracellular caveolin-1 distribution in the suppression of antigen-induced mast cell activation by cationic liposomes.

    Inoh, Yoshikazu / Tsuchiya, Yuuki / Nakanishi, Yokiko / Yokawa, Satoru / Furuno, Tadahide

    Cell biology international

    2020  Volume 44, Issue 4, Page(s) 1068–1075

    Abstract: Cationic liposomes are commonly used as vectors to effectively introduce foreign genes into target cells. In another function, we recently showed that cationic liposomes bound to the mast cell surface suppress the degranulation induced by the cross- ... ...

    Abstract Cationic liposomes are commonly used as vectors to effectively introduce foreign genes into target cells. In another function, we recently showed that cationic liposomes bound to the mast cell surface suppress the degranulation induced by the cross-linking of high-affinity immunoglobulin E receptor in a time- and dose-dependent manner. This suppression is mediated by the impairment of the sustained level of intracellular Ca
    MeSH term(s) Animals ; Calcium/metabolism ; Caveolae/metabolism ; Caveolin 1/metabolism ; Cell Line, Tumor ; Cell Membrane/metabolism ; Liposomes/metabolism ; Mast Cells/cytology ; Mast Cells/metabolism ; Phosphatidylinositol 3-Kinase/metabolism ; Rats
    Chemical Substances Caveolin 1 ; Liposomes ; Phosphatidylinositol 3-Kinase (EC 2.7.1.137) ; Calcium (SY7Q814VUP)
    Language English
    Publishing date 2020-01-09
    Publishing country England
    Document type Journal Article
    ZDB-ID 1143453-3
    ISSN 1095-8355 ; 1065-6995
    ISSN (online) 1095-8355
    ISSN 1065-6995
    DOI 10.1002/cbin.11297
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  3. Article ; Online: Inhibition of degranulation in mast cells attached to a hydrogel through defective microtubule tracts.

    Shiki, Atsushi / Inoh, Yoshikazu / Yokawa, Satoru / Furuno, Tadahide

    Experimental cell research

    2019  Volume 381, Issue 2, Page(s) 248–255

    Abstract: Mast cells (MCs) are important effectors of the immediate allergic response. MCs are distributed throughout various tissues and organs, and adhere to extracellular matrix (ECM) with broad stiffness in the body. Here we compared cellular responses ... ...

    Abstract Mast cells (MCs) are important effectors of the immediate allergic response. MCs are distributed throughout various tissues and organs, and adhere to extracellular matrix (ECM) with broad stiffness in the body. Here we compared cellular responses following antigen stimulation in MCs on glass-base dishes with and without a hydrogel. We found that an antigen-induced increase in intracellular Ca
    MeSH term(s) Animals ; Calcium/metabolism ; Cell Adhesion/drug effects ; Cell Culture Techniques/instrumentation ; Cell Culture Techniques/methods ; Cell Degranulation/drug effects ; Cell Degranulation/physiology ; Cell Line ; Extracellular Matrix/drug effects ; Extracellular Matrix/metabolism ; Focal Adhesions/drug effects ; Focal Adhesions/metabolism ; Hydrogels/chemistry ; Hydrogels/pharmacology ; Mast Cells/cytology ; Mast Cells/drug effects ; Mast Cells/physiology ; Microtubules/drug effects ; Microtubules/metabolism ; Microtubules/pathology ; Rats ; Signal Transduction/drug effects ; Surface Properties ; Tissue Scaffolds/chemistry
    Chemical Substances Hydrogels ; Calcium (SY7Q814VUP)
    Language English
    Publishing date 2019-05-18
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 1493-x
    ISSN 1090-2422 ; 0014-4827
    ISSN (online) 1090-2422
    ISSN 0014-4827
    DOI 10.1016/j.yexcr.2019.05.019
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    Zs.A 563: Show issues Location:
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  4. Article ; Online: Effects of lipid composition in cationic liposomes on suppression of mast cell activation.

    Inoh, Yoshikazu / Hirose, Takuya / Yokoi, Asami / Yokawa, Satoru / Furuno, Tadahide

    Chemistry and physics of lipids

    2020  Volume 231, Page(s) 104948

    Abstract: We previously showed that cationic liposomes composed of cholesteryl-3β-carboxyamidoethylene-N-hydroxyethylamine (OH-Chol) and 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) inhibited mast cell degranulation mediated by the cross-linking of ... ...

    Abstract We previously showed that cationic liposomes composed of cholesteryl-3β-carboxyamidoethylene-N-hydroxyethylamine (OH-Chol) and 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) inhibited mast cell degranulation mediated by the cross-linking of high-affinity IgE receptors (FcεRI). In this study, we prepared three kinds of cationic liposomes composed of OH-Chol and DOPE in different ratios (0.28, 0.60, and 0.86 of OH-Chol in mol ratio, named as L-liposome, M-liposome, and H-liposome, respectively) and investigated their effects on mast cell activation. We found that mast cell degranulation evoked with antigen was inhibited by pretreatment with cationic liposomes in the composite ratio-dependent manner of OH-Chol and that the H-liposome showed the highest inhibitory effect on degranulation among three kinds of liposomes. Store-operated Ca
    MeSH term(s) Animals ; Calcium/metabolism ; Cations/chemistry ; Cells, Cultured ; Endoplasmic Reticulum/drug effects ; Endoplasmic Reticulum/metabolism ; Lipids/chemistry ; Lipids/pharmacology ; Liposomes/chemistry ; Mast Cells/drug effects ; Mast Cells/metabolism ; Rats
    Chemical Substances Cations ; Lipids ; Liposomes ; Calcium (SY7Q814VUP)
    Language English
    Publishing date 2020-07-25
    Publishing country Ireland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 213869-4
    ISSN 1873-2941 ; 0009-3084
    ISSN (online) 1873-2941
    ISSN 0009-3084
    DOI 10.1016/j.chemphyslip.2020.104948
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  5. Article ; Online: Receptor dynamics regulates actin polymerization state through phosphorylation of cofilin in mast cells.

    Suzuki, Ruriko / Inoh, Yoshikazu / Yokawa, Satoru / Furuno, Tadahide / Hirashima, Naohide

    Biochemical and biophysical research communications

    2020  Volume 534, Page(s) 714–719

    Abstract: Aggregation of IgE bound to the high-affinity IgE receptor (FcεRI) by a multivalent antigen induces mast cell activation, while disaggregation of aggregated FcεRI by monomer hapten immediately terminates degranulation mediated by dephosphorylation of Syk ...

    Abstract Aggregation of IgE bound to the high-affinity IgE receptor (FcεRI) by a multivalent antigen induces mast cell activation, while disaggregation of aggregated FcεRI by monomer hapten immediately terminates degranulation mediated by dephosphorylation of Syk and mediates a decrease in intracellular Ca
    MeSH term(s) Actin Depolymerizing Factors/metabolism ; Actins/metabolism ; Animals ; Calcium Signaling ; Cell Line ; Cytochalasin D/pharmacology ; Mast Cells/drug effects ; Mast Cells/metabolism ; Ovalbumin/pharmacology ; Phalloidine/chemistry ; Phalloidine/metabolism ; Phosphorylation ; Polymerization ; Rats ; Receptors, IgE/metabolism ; Rhodamines/chemistry ; Rhodamines/metabolism
    Chemical Substances Actin Depolymerizing Factors ; Actins ; FCER1A protein, rat ; Receptors, IgE ; Rhodamines ; trinitrophenyl-ovalbumin ; Phalloidine (17466-45-4) ; Cytochalasin D (22144-77-0) ; Ovalbumin (9006-59-1)
    Language English
    Publishing date 2020-11-17
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 205723-2
    ISSN 1090-2104 ; 0006-291X ; 0006-291X
    ISSN (online) 1090-2104 ; 0006-291X
    ISSN 0006-291X
    DOI 10.1016/j.bbrc.2020.11.012
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  6. Article ; Online: Promotion of microtubule acetylation plays an important role in degranulation of antigen-activated mast cells.

    Shiki, Atsushi / Inoh, Yoshikazu / Yokawa, Satoru / Furuno, Tadahide

    Inflammation research : official journal of the European Histamine Research Society ... [et al.

    2018  Volume 68, Issue 3, Page(s) 181–184

    Abstract: Objective: The aim of this study was to investigate whether microtubule acetylation is triggered by antigen stimulation and how it affects mast cell degranulation.: Methods: The RBL-2H3 cell line was used as a model for mast cells. Acetylation of α- ... ...

    Abstract Objective: The aim of this study was to investigate whether microtubule acetylation is triggered by antigen stimulation and how it affects mast cell degranulation.
    Methods: The RBL-2H3 cell line was used as a model for mast cells. Acetylation of α-tubulin was analyzed by Western blotting. Intracellular distribution of α-tubulin and acetylated α-tubulin was observed by immunostaining. Degranulation was monitored by measuring the activity of β-hexosaminidase secreted into cell supernatants. Tukey-Kramer test was used to compare differences between groups.
    Results: Microtubule acetylation proceeds globally in mast cell cytoplasm after antigen stimulation in addition to accelerated formation of microtubule-organizing centers. Pretreatment with 5Z-7-oxozeaenol (5 µmol/l), an inhibitor of TGF-β-activated kinase 1, which is a key activator of α-tubulin acetyltransferase 1, did not affect the distribution and acetylation of microtubules in resting cells; however, it significantly suppressed antigen-evoked microtubule acetylation and their reorganization, and subsequent degranulation (95.0 ± 1.2% inhibition, n = 3, P < 0.01).
    Conclusions: These results provided new insight into the post-translational modifications of microtubule to regulate mast cell degranulation.
    MeSH term(s) Acetylation ; Animals ; Antigens/physiology ; Cell Degranulation ; Cell Line ; Mast Cells/physiology ; Microtubules/physiology ; Rats ; Tubulin/physiology ; beta-N-Acetylhexosaminidases/metabolism
    Chemical Substances Antigens ; Tubulin ; beta-N-Acetylhexosaminidases (EC 3.2.1.52)
    Language English
    Publishing date 2018-11-23
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 1221794-3
    ISSN 1420-908X ; 1023-3830
    ISSN (online) 1420-908X
    ISSN 1023-3830
    DOI 10.1007/s00011-018-1203-2
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  7. Article ; Online: Decreased intracellular granule movement and glucagon secretion in pancreatic α cells attached to superior cervical ganglion neurites.

    Watabe, Kiyoto / Yokawa, Satoru / Inoh, Yoshikazu / Suzuki, Takahiro / Furuno, Tadahide

    Molecular and cellular biochemistry

    2018  Volume 446, Issue 1-2, Page(s) 83–89

    Abstract: Autonomic neurons innervate pancreatic islets of Langerhans and participate in the maintenance of blood glucose concentrations by controlling hormone levels through attachment with islet cells. We previously found that stimulated superior cervical ... ...

    Abstract Autonomic neurons innervate pancreatic islets of Langerhans and participate in the maintenance of blood glucose concentrations by controlling hormone levels through attachment with islet cells. We previously found that stimulated superior cervical ganglia (SCG) could induce Ca
    MeSH term(s) Animals ; Cell Line ; Coculture Techniques ; Glucagon/secretion ; Glucagon-Secreting Cells/cytology ; Glucagon-Secreting Cells/metabolism ; Mice ; Mice, Inbred BALB C ; Neurites/metabolism ; Secretory Vesicles/metabolism ; Superior Cervical Ganglion/cytology ; Superior Cervical Ganglion/metabolism
    Chemical Substances Glucagon (9007-92-5)
    Language English
    Publishing date 2018-01-09
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 184833-1
    ISSN 1573-4919 ; 0300-8177
    ISSN (online) 1573-4919
    ISSN 0300-8177
    DOI 10.1007/s11010-018-3275-2
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  8. Article ; Online: Monomer hapten and hapten-specific IgG inhibit mast cell activation evoked by multivalent hapten with different mechanisms.

    Suzuki, Ruriko / Inoh, Yoshikazu / Yokawa, Satoru / Suzuki, Ryo / Furuno, Tadahide / Hirashima, Naohide

    European journal of immunology

    2019  Volume 49, Issue 12, Page(s) 2172–2183

    Abstract: Aggregation of IgE bound to high affinity IgE receptor (FcεRI) by multivalent antigen induces mast cell activation. Reportedly, disaggregation of aggregated FcεRI immediately terminated degranulation, and formation of co-ligated FcεRI and low affinity ... ...

    Abstract Aggregation of IgE bound to high affinity IgE receptor (FcεRI) by multivalent antigen induces mast cell activation. Reportedly, disaggregation of aggregated FcεRI immediately terminated degranulation, and formation of co-ligated FcεRI and low affinity IgG receptor FcγRIIB blocked degranulation by inhibitory signal via SH2-containing inositol 5'-phosphatase 1 (SHIP1) phosphorylation. However, their molecular mechanisms to inhibit mast cell activation have been unclear in detail. Herein, we found that addition of excess monomeric hapten (TNP-alanine) to multivalent antigen (TNP-OVA)-activated rat basophilic leukemia cells and mouse bone marrow-derived mast cells induced immediate and transient Syk dephosphorylation, which was previously phosphorylated by TNP-OVA addition. Syk dephosphorylation correlated to rapidly decreased intracellular Ca
    MeSH term(s) Animals ; Antibodies, Monoclonal, Murine-Derived/immunology ; Cell Line, Tumor ; Haptens/immunology ; Immunoglobulin G/immunology ; Immunologic Capping/immunology ; Mast Cells/cytology ; Mast Cells/immunology ; Mice ; Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases/immunology ; Rats ; Receptors, IgE/immunology ; Receptors, IgG/immunology
    Chemical Substances Antibodies, Monoclonal, Murine-Derived ; Fcgr2b protein, mouse ; Fcgr2b protein, rat ; Haptens ; Immunoglobulin G ; Receptors, IgE ; Receptors, IgG ; Inpp5d protein, mouse (EC 3.1.3.86) ; Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases (EC 3.1.3.86)
    Language English
    Publishing date 2019-08-13
    Publishing country Germany
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 120108-6
    ISSN 1521-4141 ; 0014-2980
    ISSN (online) 1521-4141
    ISSN 0014-2980
    DOI 10.1002/eji.201847973
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    Zs.MG 85: Show issues

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  9. Article ; Online: Visualization of osteocalcin and bone morphogenetic protein 2 (BMP2) secretion from osteoblastic cells by bioluminescence imaging.

    Sugimoto, Honami / Fukuda, Shinji / Yokawa, Satoru / Hori, Miki / Ninomiya, Hotsuna / Sato, Takuma / Miyazawa, Ken / Kawai, Tatsushi / Furuno, Tadahide / Inouye, Satoshi / Goto, Shigemi / Suzuki, Takahiro

    Biochemical and biophysical research communications

    2022  Volume 635, Page(s) 203–209

    Abstract: The secretions of osteocalcin and bone morphogenetic protein 2 (BMP2) from living osteoblastic cells were visualized for the first time using a method of video-rate bioluminescence imaging. The fusion proteins with Gaussia luciferase (GLase) for mouse ... ...

    Abstract The secretions of osteocalcin and bone morphogenetic protein 2 (BMP2) from living osteoblastic cells were visualized for the first time using a method of video-rate bioluminescence imaging. The fusion proteins with Gaussia luciferase (GLase) for mouse osteocalcin and BMP2 (OC-GLase and BMP2-GLase, respectively) expressed in osteoblastic MC3T3-E1 cells were correctly processed and secreted. In the video images of exocytotic secretion, the luminescence spots of OC-GLase and BMP2-GLase disappeared rapidly and gradually, respectively, indicating different manners of these proteins in diffusion. Notably, a deletion mutant of BMP2 (Δ3BMP2-GLase) lacking three basic amino acid residues in the N-terminal region for binding to heparan sulfate showed rapidly disappearing luminescence spots. In our imaging conditions, the half-life of luminescence for the spots of Δ3BMP2-GLase (1.61 ± 0.20 s) was similar to that of OC-GLase (1.22 ± 0.14 s) but not to that of BMP2-GLase (4.31 ± 0.41 s). These results suggest that, in contrast to osteocalcin, the diffusion of BMP2 from cells occurred slowly after exocytosis. Thus, our bioluminescence imaging method is useful to study the diffusion properties of secreted proteins in exocytosis.
    MeSH term(s) Mice ; Animals ; Osteocalcin/genetics ; Osteocalcin/metabolism ; Bone Morphogenetic Protein 2/metabolism ; Luciferases/genetics ; Luciferases/metabolism ; Cell Communication ; Cell Line ; Osteoblasts/metabolism ; Cell Differentiation
    Chemical Substances Osteocalcin (104982-03-8) ; Bone Morphogenetic Protein 2 ; Luciferases (EC 1.13.12.-) ; Bmp2 protein, mouse
    Language English
    Publishing date 2022-10-14
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 205723-2
    ISSN 1090-2104 ; 0006-291X ; 0006-291X
    ISSN (online) 1090-2104 ; 0006-291X
    ISSN 0006-291X
    DOI 10.1016/j.bbrc.2022.10.042
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  10. Article: Cationic liposomes suppress intracellular calcium ion concentration increase via inhibition of PI3 kinase pathway in mast cells.

    Inoh, Yoshikazu / Haneda, Aki / Tadokoro, Satoshi / Yokawa, Satoru / Furuno, Tadahide

    Biochimica et biophysica acta. Biomembranes

    2017  Volume 1859, Issue 12, Page(s) 2461–2466

    Abstract: Cationic liposomes are commonly used as vectors to effectively introduce foreign genes (antisense DNA, plasmid DNA, siRNA, etc.) into target cells. Cationic liposomes are also known to affect cellular immunocompetences such as the mast cell function in ... ...

    Abstract Cationic liposomes are commonly used as vectors to effectively introduce foreign genes (antisense DNA, plasmid DNA, siRNA, etc.) into target cells. Cationic liposomes are also known to affect cellular immunocompetences such as the mast cell function in allergic reactions. In particular, we previously showed that the cationic liposomes bound to the mast cell surface suppress the degranulation induced by cross-linking of high affinity IgE receptors in a time- and dose-dependent manner. This suppression is mediated by impairment of the sustained level of intracellular Ca
    MeSH term(s) Animals ; Calcium/metabolism ; Calcium Channels/genetics ; Calcium Channels/metabolism ; Calcium Signaling ; Cations ; Cell Line ; Cell Membrane/drug effects ; Cell Membrane/metabolism ; Cholesterol/analogs & derivatives ; Cholesterol/chemistry ; Endoplasmic Reticulum/drug effects ; Endoplasmic Reticulum/metabolism ; Gene Expression Regulation ; Ion Transport/drug effects ; Liposomes/chemistry ; Liposomes/pharmacology ; Mast Cells/cytology ; Mast Cells/drug effects ; Mast Cells/metabolism ; ORAI1 Protein/genetics ; ORAI1 Protein/metabolism ; Phosphatidylethanolamines/chemistry ; Phosphatidylinositol 3-Kinase/antagonists & inhibitors ; Phosphatidylinositol 3-Kinase/genetics ; Phosphatidylinositol 3-Kinase/metabolism ; Phosphorylation/drug effects ; Proto-Oncogene Proteins c-akt/antagonists & inhibitors ; Proto-Oncogene Proteins c-akt/genetics ; Proto-Oncogene Proteins c-akt/metabolism ; Rats ; Stromal Interaction Molecule 1/genetics ; Stromal Interaction Molecule 1/metabolism
    Chemical Substances Calcium Channels ; Cations ; Liposomes ; ORAI1 Protein ; Orai1 protein, rat ; Phosphatidylethanolamines ; Stim1 protein, rat ; Stromal Interaction Molecule 1 ; cholesteryl-3-carboxyamidoethylene-N-hydroxyethylamine ; dioleoyl phosphatidylethanolamine (2462-63-7) ; Cholesterol (97C5T2UQ7J) ; Phosphatidylinositol 3-Kinase (EC 2.7.1.137) ; Proto-Oncogene Proteins c-akt (EC 2.7.11.1) ; Calcium (SY7Q814VUP)
    Language English
    Publishing date 2017-09-29
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 60-7
    ISSN 1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0005-2736 ; 0006-3002 ; 0005-2728 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
    ISSN (online) 1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650
    ISSN 0005-2736 ; 0006-3002 ; 0005-2728 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
    DOI 10.1016/j.bbamem.2017.09.025
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