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  • 1. Antunes, Fernando
    et al.
    Cadenas, Enrique
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Apoptosis, induced by exposure to a low and steady-state concentration of H2O2, is a consequence of lysosomal rupture2001In: Biochemical Journal, ISSN 0264-6021, E-ISSN 1470-8728, Vol. 356, p. 549-555Article in journal (Refereed)
  • 2.
    Autelli, Riccardo
    et al.
    Department of Experimental Medicine and Oncology, University of Turin, Italy.
    Ullio, Chiara
    Department of Experimental Medicine and Oncology, University of Turin, Italy.
    Prigione, Elisa
    Department of Experimental Medicine and Oncology, University of Turin, Italy.
    Schiavone, Nicola
    Department of Experimental Medicine and Oncology, University of Florence, Italy.
    Brunk, Ulf
    Linköping University, Department of Medicine and Care, Pharmacology. Linköping University, Faculty of Health Sciences.
    Capaccioli, Sergio
    Department of Experimental Medicine and Oncology, University of Florence, Italy.
    Baccino, Francesco
    Department of Experimental Medicine and Oncology, University of Turin, Italy.
    Bonelli, Gabriella
    Department of Experimental Medicine and Oncology, University of Turin, Italy.
    Divergent pathways for TNF and C₂-ceramide toxicity in HTC hematoma cells2009In: Biochimica et Biophysica Acta, ISSN 0006-3002, E-ISSN 1878-2434, Vol. 1793, p. 1182-1190Article in journal (Refereed)
    Abstract [en]

    We previously showed that, in the rat hepatoma cell line HTC, TNF brings about a non-caspase-dependent, apoptosis-like process requiring NADPH oxidase activity, an iron-mediated pro-oxidant status, and a functional acidic vacuolar compartment. This process may thus involve mechanisms such as autophagy or relocation of lysosomal enzymes, perhaps secondary to the formation of ceramide by acidic sphingomyelinase. Here we investigated whether ceramide formation contributes to the apoptogenic process. HTC cells were found to be sensitive to exogenous ceramide and significantly protected against TNF by desipramine, an inhibitor of lysosomal acid sphingomyelinase. However, Bcl-2 transfection and Bcl-x(L) upregulation by dexamethasone significantly diminished the apoptogenic effect of ceramide but not that of TNF, suggesting that ceramide is not directly involved in TNF toxicity. Moreover, Bcl-x(L) silencing precluded dexamethasone-induced protection against ceramide and, by itself, induced massive death, demonstrating the strict dependence of HTC cells on Bcl-x(L) for survival also under standard culture conditions.

  • 3. Baird, Sarah K
    et al.
    Kurz, Tino
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Metallothionein protects against oxidative stress-induced lysosomal destabilization2006In: Biochemical Journal, ISSN 0264-6021, E-ISSN 1470-8728, Vol. 394, no 1, p. 275-283Article in journal (Refereed)
    Abstract [en]

    The introduction of apo-ferritin or the iron chelator DFO (desferrioxamine) conjugated to starch into the lysosomal compartment protects cells against oxidative stress, lysosomal rupture and ensuing apoptosis/necrosis by binding intralysosomal redox-active iron, thus preventing Fenton-type reactions and ensuing peroxidation of lysosomal membranes. Because up-regulation of MTs (metallothioneins) also generates enhanced cellular resistance to oxidative stress, including X-irradiation, and MTs were found to be capable of iron binding in an acidic and reducing lysosomal-like environment, we propose that these proteins might similarly stabilize lysosomes following autophagocytotic delivery to the lysosomal compartment. Here, we report that Zn-mediated MT up-regulation, assayed by Western blotting and immunocytochemistry, results in lysosomal stabilization and decreased apoptosis following oxidative stress, similar to the protection afforded by fluid-phase endocytosis of apo-ferritin or DFO. In contrast, the endocytotic uptake of an iron phosphate complex destabilized lysosomes against oxidative stress, but this was suppressed in cells with up-regulated MT. It is suggested that the resistance against oxidative stress, known to occur in MT-rich cells, may be a consequence of autophagic turnover of MT, resulting in reduced iron-catalysed intralysosomal peroxidative reactions. © 2006 Biochemical Society.

  • 4.
    Berndt, Carsten
    et al.
    Division of Biochemistry, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, Sweden; Institute for Clinical Cytobiology and Cytopathology, Philipps-Universität, 35037 Marburg, Germany..
    Kurz, Tino
    Linköping University, Department of Medical and Health Sciences, Clinical Pharmacology. Linköping University, Faculty of Health Sciences.
    Bannenberg, Sarah
    Division of Biochemistry, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, Sweden.
    Jacob, Ralf
    Institute for Clinical Cytobiology and Cytopathology, Philipps-Universität, 35037 Marburg, Germany..
    Holmgren, Arne
    Division of Biochemistry, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, Sweden.
    Brunk, Ulf T
    Linköping University, Department of Medical and Health Sciences, Clinical Pharmacology. Linköping University, Faculty of Health Sciences.
    Ascorbate and endocytosed Motexafin gadolinium induce lysosomal rupture.2011In: Cancer Letters, ISSN 0304-3835, E-ISSN 1872-7980, Vol. 307, no 2, p. 119-23Article in journal (Refereed)
    Abstract [en]

    Motexafin gadolinium (MGd) sensitizes malignant cells to ionizing radiation, although the underlying mechanisms for uptake and sensitization are both unclear. Here we show that MGd is endocytosed by the clathrin-dependent pathway with ensuing lysosomal membrane permeabilization, most likely via formation of reactive oxygen species involving redox-active metabolites, such as ascorbate. We propose that subsequent apoptosis is a synergistic effect of irradiation and high MGd concentrations in malignant cells due to their pronounced endocytic activity. The results provide novel insights into the mode of action of this promising anti-cancer drug, which is currently under clinical trials.

  • 5.
    Berndt, Carsten
    et al.
    Division for Biochemistry, Department for Medical Biochemistry and Biophysics, Karolinska Institute, Sweden. Institute for Cinical Cytobiology and Cytopathology, Philipps-Universität, Marburg, Germany .
    Kurz, Tino
    Linköping University, Department of Medicine and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Selenius, Markus
    Division of Pathology, Department of Laboratory Medicineand Division of Medical Radiation Physics, Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden.
    Fernandes, Aristi P
    Division of Pathology, Department of Laboratory Medicineand Division of Medical Radiation Physics, Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden.
    Edgren, Margareta R
    Division of Medical Radiation Physics, Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden.
    Brunk, Ulf T
    Linköping University, Department of Medicine and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Chelation of lysosomal iron protects against ionizing radiation.2010In: Biochemical Journal, ISSN 0264-6021, E-ISSN 1470-8728, Vol. 432, no 2, p. 295-301Article in journal (Refereed)
    Abstract [en]

    Ionizing radiation causes DNA damage and consequent apoptosis, mainly due to the production of hydroxyl radicals (HO•) that follows radiolytic splitting of water. However, superoxide (O2•-) and H2O2 also form and induce oxidative stress with resulting LMP (lysosomal membrane permeabilization) arising from iron-catalysed oxidative events. The latter will contribute significantly to radiation-induced cell death and its degree largely depends on the quantities of lysosomal redox-active iron present as a consequence of autophagy and endocytosis of iron-rich compounds. Therefore radiation sensitivity might be depressed by lysosome-targeted iron chelators. In the present study, we have shown that cells in culture are significantly protected from ionizing radiation damage if initially exposed to the lipophilic iron chelator SIH (salicylaldehyde isonicotinoyl hydrazone), and that this effect is based on SIH-dependent lysosomal stabilization against oxidative stress. According to its dose-response-modifying effect, SIH is a most powerful radioprotector and a promising candidate for clinical application, mainly to reduce the radiation sensitivity of normal tissue. We propose, as an example, that inhalation of SIH before each irradiation session by patients undergoing treatment for lung malignancies would protect normally aerated lung tissue against life-threatening pulmonary fibrosis, whereas the sensitivity of malignant lung tumours, which usually are non-aerated, will not be affected by inhaled SIH.

  • 6.
    Bironaite, Daiva
    et al.
    State Research Institute Centre Innovat Med, Lithuania .
    Brunk, Ulf
    Linköping University, Department of Medical and Health Sciences, Division of Drug Research. Linköping University, Faculty of Health Sciences.
    Venalis, Algirdas
    State Research Institute Centre Innovat Med, Lithuania .
    Protective Induction of Hsp70 in Heat-Stressed Primary Myoblasts: Involvement of MAPKs2013In: Journal of Cellular Biochemistry, ISSN 0730-2312, E-ISSN 1097-4644, Vol. 114, no 9, p. 2024-2031Article in journal (Refereed)
    Abstract [en]

    The involvement of extracellular signal-regulated kinases 1 and 2 (ERK1,2), stress kinase p38 and c-Jun NH2-terminal kinases 1 and 2 (JNK1,2) on Hsp70-upregulation following mild heat shock, and resulting cell protection, was studied on rabbit primary myoblasts. Cells subjected to heat stress (42 degrees C; 60min) showed a significantly enhanced amount of heat-shock-induced protein 70 (Hsp70), correlating with sustained phosphorylation of MAP kinases ERK1,2, inhibition of p38 and JNK1,2 activation. Induced Hsp70 did not autocrinally suppress activation of transcription factor c-Jun, suggesting involvement of the latter in the protection of myoblasts following heat shock. The inhibition of stress kinases p38, JNK1,2, and MEK1,2 by SP600125, SB203580, and UO126, respectively, established the involvement of JNK1,2 and p38 as upstream, and ERK1,2 as downstream targets of Hsp70 induction. Moreover, the effect of the MEK1,2 inhibitor UO126 revealed a new pathway of c-Jun activation by ERK1,2 in myogenic heat-stressed stem cells. The presented data show that transient activation of JNK1, JNK2, and p38 is necessary for Hsp70 induction and ensuing cell protection. In conclusion, affecting myogenic stem cell protective mechanisms might be a useful strategy in improving stem cell survival and their expanded application in therapy.

  • 7.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Book review "Lysosomal pathways of protein degradation"2001In: Experimental Gerontology, ISSN 0531-5565, E-ISSN 1873-6815, Vol. 36, p. 1419-1420Article in journal (Refereed)
  • 8.
    Brunk, Ulf
    Linköping University, Department of Medicine and Care, Pharmacology. Linköping University, Faculty of Health Sciences.
    Growing cells at 40% ambient oxygen conditions them to subsequent oxidative stress2007In: Biogerontology (Dordrecht), ISSN 1389-5729, E-ISSN 1573-6768, Vol. 8, no 5, p. 619-620Article in journal (Other academic)
  • 9.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    Lysosomal involvement in apoptosis2002In: Free Radical Biology & Medicine, ISSN 0891-5849, E-ISSN 1873-4596, Vol. 33, p. 195-Conference paper (Other academic)
  • 10.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Lysosomotropic detergents induce time- and dose- dependent apoptosis/necrosis in cultured cells.2000In: Redox report, ISSN 1351-0002, E-ISSN 1743-2928, Vol. 5, p. 87-88Article in journal (Refereed)
  • 11.
    Brunk, Ulf
    et al.
    Linköping University, Department of Medicine and Care, Pharmacology. Linköping University, Faculty of Health Sciences.
    Eaton, John W
    Molecular Targets Program, James Graham Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA.
    Commentary: Peroxide hormesis? A commentary on "Hydrogen peroxide inhibits caspase-dependent apoptosis by inactivating procaspase-9 in an iron-dependent manner": Refers to: Alexandra Barbouti, Christos Amorgianiotis, Evangelos Kolettas, Panagiotis Kanavaros, Dimitrios Galaris Hydrogen peroxide inhibits caspase-dependent apoptosis by inactivating procaspase-9 in an iron-dependent mannerFree Radical Biology and Medicine, Volume 43, Issue 10, 15 November 2007, Pages 1377-13872007In: Free Radical Biology and Medicine, ISSN 0891-5849, Vol. 43, no 10, p. 1372-1373Article in journal (Other academic)
    Abstract [en]

    [No abstract available]

  • 12.
    Brunk, Ulf
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    Neuzil, J.
    Institute for Prevention of Cardiovascular Diseases, Ludwig Maximilians University, Munich, Germany.
    Eaton, J.W.
    James Graham Brown Cancer Center, University of Louisville, Louisville, KY, United States.
    Lysosomal involvement in apoptosis2001In: Redox report, ISSN 1351-0002, E-ISSN 1743-2928, Vol. 6, no 2, p. 91-97Article, review/survey (Refereed)
    Abstract [en]

    [No abstract available]

  • 13.
    Brunk, Ulf
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Svensson, Irene
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Oxidative stress, growth-factor starvation, and CD-95-activation may all cause apoptosis through lysosomal leak.1999In: Redox report, ISSN 1351-0002, E-ISSN 1743-2928, Vol. 4Article in journal (Refereed)
  • 14.
    Brunk, Ulf
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Terman, Alexei
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology.
    Lipofuscin: Mechanisms of age-related accumulation and influence on cell function2002In: Free Radical Biology & Medicine, ISSN 0891-5849, E-ISSN 1873-4596, Vol. 33, no 5, p. 611-619Article in journal (Refereed)
    Abstract [en]

    The accumulation of lipofuscin within postmitotic cells is a recognized hallmark of aging occuring with a rate inversely related to longevity. Lipofuscin is an intralysosomal, polymeric substance, primarily composed of cross-linked protein residues, formed due to iron-catalyzed oxidative processes. Because it is undegradable and cannot be removed via exocytosis, lipofuscin accumulation in postmitotic cells is inevitable, whereas proliferative cells efficiently dilute it during division. The rate of lipofuscin formation can be experimentally manipulated. In cell culture models, oxidative stress (e.g., exposure to 40% ambient oxygen or low molecular weight iron) promotes lipofuscin accumulation, whereas growth at 8% oxygen and treatment with antioxidants or iron-chelators diminish it. Lipofuscin is a fluorochrome and may sensitize lysosomes to visible light, a process potentially important for the pathogenesis of age-related macular degeneration. Lipofuscin-associated iron sensitizes lysosomes to oxidative stress, jeopardizing lysosomal stability and causing apoptosis due to release of lysosomal contents. Lipofuscin accumulation may also diminish autophagocytotic capacity by acting as a sink for newly produced lysosomal enzymes and, therefore, interfere with recycling of cellular components. Lipofuscin, thus, may be much more directly related to cellular degeneration at old age than was hitherto believed.

  • 15.
    Brunk, Ulf
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Terman, Alexei
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology.
    The mitochondrial-lysosomal axis theory of aging: Accumulation of damaged mitochondria as a result of imperfect autophagocytosis2002In: European Journal of Biochemistry, ISSN 0014-2956, E-ISSN 1432-1033, Vol. 269, no 8, p. 1996-2002Article in journal (Refereed)
    Abstract [en]

    Cellular manifestations of aging are most pronounced in postmitotic cells, such as neurons and cardiac myocytes. Alterations of these cells, which are responsible for essential functions of brain and heart, are particularly important contributors to the overall aging process. Mitochondria and lysosomes of postmitotic cells suffer the most remarkable age-related alterations of all cellular organelles. Many mitochondria undergo enlargement and structural disorganization, while lysosomes, which are normally responsible for mitochondrial turnover, gradually accumulate an undegradable, polymeric, autofluorescent material called lipofuscin, or age pigment. We believe that these changes occur not only due to continuous oxidative stress (causing oxidation of mitochondrial constituents and autophagocytosed material), but also because of the inherent inability of cells to completely remove oxidatively damaged structures (biological 'garbage'). A possible factor limiting the effectiveness of mitochondial turnover is the enlargement of mitochondria which may reflect their impaired fission. Non-autophagocytosed mitochondria undergo further oxidative damage, resulting in decreasing energy production and increasing generation of reactive oxygen species. Damaged, enlarged and functionally disabled mitochondria gradually displace normal ones, which cannot replicate indefinitely because of limited cell volume. Although lipofuscin-loaded lysosomes continue to receive newly synthesized lysosomal enzymes, the pigment is undegradable. Therefore, advanced lipofuscin accumulation may greatly diminish lysosomal degradative capacity by preventing lysosomal enzymes from targeting to functional autophagosomes, further limiting mitochondrial recycling. This interrelated mitochondrial and lysosomal damage irreversibly leads to functional decay and death of postmitotic cells.

  • 16.
    Brunk, Ulf
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    Yu, ZQ
    Persson, Lennart
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Experimental Pathology . Östergötlands Läns Landsting, Centre of Surgery and Oncology, Department of Respiratory Medicine UHL.
    Eaton, John Wallace
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Experimental Pathology . Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Lysosomes, iron and oxidative stress2003In: Free radical research, ISSN 1071-5762, E-ISSN 1029-2470, Vol. 37, p. 34-34Conference paper (Other academic)
  • 17.
    Brunk, Ulf
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Zhao, Ming
    Experimental Vascular Research Unit, Clinical Research Center, Lund University.
    Septic shock and the lysosomal-mitochondrial axis theory of apoptosis2009In: Molecular Mechanism of Severe Shock, Kerala, India: Research Signpost , 2009, p. 91-106Chapter in book (Other academic)
    Abstract [en]

    Over 250 years ago, Le Dran, a distinguished French Surgeon, first used the word “shock” in his treatise. Since then, much progress has been made in shock research. However, many questions have still confused the medical doctors until now. For example, why the reduced blood pressure can’t return to normal after anti-shock treatment in severe shock, why the high mortality of septic shock can’t be reduced though many new therapies are reported, what is the reason for the development of systemic inflammatory response syndrome and multiple organ dysfunction syndrome following a prolonged severe shock, and what event triggers the connection among shock, systemic inflammation, and multi-organ dysfunction, etc. In order to resolve these questions, research on the pathogenesis of shock has been made at molecular level in recent years. Current advances of shock molecular mechanism are presented in this book, which is divided into 10 chapters, including new theory about auto-digestion in shock and organ failure, septic shock and the lysosomal-mitochondrial axis theory in apoptosis, molecular mechanism of endotoxin action, HMGB-1 and sepsis after burns, role of MAPK in inflammation and septic shock, ion channels and low vasoreactivity in severe shock, vascular permeability in shock, lymphatic microcirculation and shock, calcium signaling in cardiac dysfunction of burns, the effect and mechanism of a new anti-shock medicine Polydatin. We hope that these chapters will help the readers to develop strategies and tactics that will promote shock research. We want to thank all the authors for their excellent cooperation and manuscript preparation. We also give special thanks to Dr. Pandalai for inviting us to edit and publish this review book in Research Signpost

     

  • 18.
    Cuervo, Ana Maria
    et al.
    Albert Einstein College of Medicine.
    Bergamini, Ettore
    University of Pisa.
    Brunk, Ulf
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Droge, Wolf
    Heidelberg, Germany.
    Ffrench, Martine
    Central Hospital, Lyon.
    Terman, Alexei
    Linköping University, Department of Clinical and Experimental Medicine, Geriatric . Linköping University, Faculty of Health Sciences.
    Autophagy and aging - The importance of maintaining "clean" cells2005In: autophagy, Vol. 1, no 3, p. 131-140Article, review/survey (Refereed)
    Abstract [en]

    A decrease in the turnover of cellular components and the intracellular accumulation of altered macromolecules and organelles are features common to all aged cells. Diminished autophagic activity plays a major role in these age-related manifestations. In this work we review the molecular defects responsible for the malfunctioning of two forms of autophagy, macroautophagy and chaperone-mediated outophagy, in old mammals, and highlight general and cell-type specific consequences of dysfunction of the autophagic system with age. Dietary caloric restriction and antilipolytic agents have been proven to efficiently stimulate autophagy in old rodents. These and other possible experimental restorative efforts are discussed.

  • 19.
    Dong, Lan-Feng
    et al.
    Griffith University.
    Freeman, Ruth
    Griffith University.
    Liu, Ji
    Griffith University.
    Zobalova, Renata
    Griffith University.
    Marin-Hernandez, Alvaro
    National Institute of Cardiology, Mexico City.
    Stantic, Marina
    Griffith University.
    Rohlena, Jakub
    Acadamy of Science, Czech Republic.
    Valis, Karel
    Acadamy of Science, Czech Republic.
    Rodriguez-Enriquez, Sara
    National Institute of Cardiology, Mexico City.
    Butcher, Bevan
    Griffith University.
    Goodwin, Jacob
    Griffith University.
    Brunk, Ulf
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Witting, Paul K
    ANZAC Research Institute.
    Moreno-Sanchez, Rafael
    National Institute of Cardiology, Mexico City.
    Scheffler, Immo E
    University California at San Diego.
    Ralph, Stephen J
    Griffith University.
    Neuzil , Jiri
    Griffith University.
    Suppression of Tumor Growth In vivo by the Mitocan alpha-tocopheryl Succinate Requires Respiratory Complex II2009In: CLINICAL CANCER RESEARCH, ISSN 1078-0432 , Vol. 15, no 5, p. 1593-1600Article in journal (Refereed)
    Abstract [en]

    Purpose: Vitamin E analogues are potent novel anticancer drugs. The purpose of this study was to elucidate the cellular target by which these agents, represented by alpha-tocopoheryl succinate (alpha-TOS), suppress tumors in vivo, with the focus on the mitochondrial complex II (CII).

    Experimental Design: Chinese hamster lung fibroblasts with functional, dysfunctional, and reconstituted CII were transformed using H-Ras. The cells were then used to form xenografts in immunocompromized mice, and response of the cells and the tumors to alpha-TOS was studied.

    Results: The CII-functional and CII-reconstituted cells, unlike their CII-dysfunctional counterparts, responded to alpha-TOS by reactive oxygen species generation and apoptosis execution. Tumors derived from these cell lines reciprocated their responses to alpha-TOS. Thus, growth of CII-functional and CII-reconstituted tumors was strongly suppressed by the agent, and this was accompanied by high level of apoptosis induction in the tumor cells. On the other hand, alpha-TOS did not inhibit the CII-dysfuntional tumors.

    Conclusions: We document in this report a novel paradigm, according to which the mitochondrial CII, which rarely mutates in human neoplasias, is a plausible target for anticancer drugs from the group of vitamin E analogues, providing support for their testing in clinical trials.

  • 20.
    Double, K L
    et al.
    Australien.
    Dedov, V N
    Australien.
    Fedorow, H
    Australien.
    Kettle, E
    Australien.
    Halliday, G M
    Australien.
    Garner, B
    Australien.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    The comparative biology of neuromelanin and lipofuscin in the human brain2008In: Cellular and Molecular Life Sciences (CMLS), ISSN 1420-682X, E-ISSN 1420-9071, Vol. 65, no 11, p. 1669-1682Article in journal (Refereed)
    Abstract [en]

    Neuromelanin and lipofuscin are two pigments produced within the human brain that, until recently, were considered inert cellular waste products of little interest to neuroscience. Recent research has increased our understanding of the nature and interactions of these pigments with their cellular environment and suggests that these pigments may, indeed, influence cellular function. The physical appearance and distribution of the pigments within the human brain differ, but both accumulate in the aging brain and the pigments share some structural features. Lipofuscin accumulation has been implicated in postmitotic cell aging, while neuromelanin is suggested to function as an iron-regulatory molecule with possible protective functions within the cells which produce this pigment. This review presents comparative aspects of the biology of neuromelanin and lipofuscin, as well as a discussion of their hypothesized functions in brain and their possible roles in aging and neurodegenerative disease. © 2008 Birkhaueser.

  • 21. Doulias, Paschalis-Thomas
    et al.
    Christoforidis, Savas
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Galaris, Dimitrios
    Endosomal and lysosomal effects of desferrioxamine: Protection of HeLa cells from hydrogen peroxide-induced DNA damage and induction of cell-cycle arrest2003In: Free Radical Biology & Medicine, ISSN 0891-5849, E-ISSN 1873-4596, Vol. 35, no 7, p. 719-728Article in journal (Refereed)
    Abstract [en]

    The role of endosomal/lysosomal redox-active iron in H2O 2-induced nuclear DNA damage as well as in cell proliferation was examined using the iron chelator desferrioxamine (DFO). Transient transfections of HeLa cells with vectors encoding dominant proteins involved in the regulation of various routes of endocytosis (dynamin and Rab5) were used to show that DFO (a potent and rather specific iron chelator) enters cells by fluid-phase endocytosis and exerts its effects by chelating redox-active iron present in the endosomal/lysosomal compartment. Endocytosed DFO effectively protected cells against H2O2-induced DNA damage, indicating the importance of endosomal/lysosomal redox-active iron in these processes. Moreover, exposure of cells to DFO in a range of concentrations (0.1 to 100 ╡M) inhibited cell proliferation in a fluid-phase endocytosis- dependent manner. Flow cytometric analysis of cells exposed to 100 ╡M DFO for 24 h showed that the cell cycle was transiently interrupted at the G 2/M phase, while treatment for 48 h led to permanent cell arrest. Collectively, the above results clearly indicate that DFO has to be endocytosed by the fluid-phase pathway to protect cells against H2O 2-induced DNA damage. Moreover, chelation of iron in the endosomal/lysosomal cell compartment leads to cell cycle interruption, indicating that all cellular labile iron is propagated through this compartment before its anabolic use is possible.

  • 22.
    Douoalis, Paschalis-Thomas
    et al.
    Ioannina Greece.
    Kotoglou, Plychronis
    Ioannina Greece.
    Tenopoulou, Margarita
    Ioannina Greece.
    Keramisanou, Dimitra
    Ioannina Greece.
    Tzavaras, Theodore
    Ioannina Greece.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Galaris, Dimitrios
    Ioannina Greece.
    Angelidis, Charalampos
    Ioannina Greece.
    Involvement of heat shock protein-70 in the mechanism of hydrogen peroxide-induced DNA damage: The role of lysosomes and iron2007In: Free Radical Biology & Medicine, ISSN 0891-5849, E-ISSN 1873-4596, Vol. 42, no 4, p. 567-577Article in journal (Refereed)
    Abstract [en]

    Heat shock protein-70 (Hsp70) is the main heat-inducible member of the 70-kDa family of chaperones that assist cells in maintaining proteins functional under stressful conditions. In the present investigation, the role of Hsp70 in the molecular mechanism of hydrogen peroxide-induced DNA damage to HeLa cells in culture was examined. Stably transfected HeLa cell lines, overexpressing or lacking Hsp70, were created by utilizing constitutive expression of plasmids containing the functional hsp70 gene or hsp70-siRNA, respectively. Compared to control cells, the Hsp70-overexpressing ones were significantly resistant to hydrogen peroxide-induced DNA damage, while Hsp70-depleted cells showed an enhanced sensitivity. In addition, the "intracellular calcein-chelatable iron pool" was determined in the presence or absence of Hsp70 and found to be related to the sensitivity of nuclear DNA to H2O2. It seems likely that the main action of Hsp70, at least in this system, is exerted at the lysosomal level, by protecting the membranes of these organelles against oxidative stress-induced destabilization. Apart from shedding additional light on the mechanistic details behind the action of Hsp70 during oxidative stress, our results indicate that modulation of cellular Hsp70 may represent a way to make cancer cells more sensitive to normal host defense mechanisms or chemotherapeutic drug treatment. © 2006 Elsevier Inc. All rights reserved.

  • 23.
    Dunlop, Rachael A
    et al.
    Heart Research Institute, Australia.
    Brunk, Ulf
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Rodgers , Kenneth J
    Heart Research Institute, Australia.
    Oxidized Proteins: Mechanisms of Removal and Consequences of Accumulation2009In: IUBMB LIFE, ISSN 1521-6543 , Vol. 61, no 5, p. 522-527Article in journal (Refereed)
    Abstract [en]

    Elevated levels of oxidized proteins are reported in diseased tissue from age-related pathologies such as atherosclerosis, neurodegenerative disorders, and cataract. Unlike the precise mechanisms that exist For the repair of nucleic acids, lipids, and carbohydrates, the primary pathway for the repair of oxidized proteins is complete catabolism to their constitutive amino acids. This process can be inefficient as is evidenced by their accumulation. It is generally considered that damaged proteins are degraded by the proteasome, however, this is only true for mildly oxidized proteins, because substrates must be unfolded to enter the narrow catalytic core. Rather, evidence suggests that moderately or heavily oxidized proteins are endocytosed and enter the endosomal/lysosomal system, indicating co-operation between the proteasomes and the lysosomes. Heavily modified substrates are incompletely degraded and accumulate within the lysosomal compartments resulting in the formation of lipofuscin-like, autofluorescent aggregates. Accumulation eventually results in impaired turnover of large organelles such as proteasomes and mitochondria, lysosomal destablization, leakage of proteases into the cytosol and apoptosis. In this review, we summarize reports published since our last assessments of the field of oxidized protein degradation including a role for modified proteins in the induction of apoptosis.

  • 24.
    Dunlop, Rachael A
    et al.
    Cell Biology Group, Heart Research Institute, Newton, NSW 2042, Australia.
    Brunk, Ulf T
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Rodgers, Kenneth J
    Cell Biology Group, Heart Research Institute, Newton, NSW 2042, Australia.
    Proteins containing oxidized amino acids induce apoptosis in human monocytes.2011In: Biochemical Journal, ISSN 0264-6021, E-ISSN 1470-8728, Vol. 435, no 1, p. 207-216Article in journal (Refereed)
    Abstract [en]

    Cellular deposits of oxidized and aggregated proteins are hallmarks of a variety of age-related disorders, but whether such proteins contribute to pathology is not well understood. We previously reported that oxidized proteins form lipofuscin/ceroid-like bodies with a lysosomal-type distribution and up-regulate the transcription and translation of proteolytic lysosomal enzymes in cultured J774 mouse macrophages. Given the recently identified role of lysosomes in the induction of apoptosis, we have extended our studies to explore a role for oxidized proteins in apoptosis. Oxidized proteins were biosynthetically generated in situ by substituting oxidized analogues for parent amino acids. Apoptosis was measured with Annexin-V/PI (propidium iodide), TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling), MMP (mitochondrial membrane permeabilization), caspase activation and cytochrome c release, and related to lysosomal membrane permeabilization. Synthesized proteins containing the tyrosine oxidation product L-DOPA (L-3,4-dihydroxyphenylalanine) were more potent inducers of apoptosis than proteins containing the phenylalanine oxidation product o-tyrosine. Apoptosis was dependent upon incorporation of oxidized residues, as indicated by complete abrogation in cultures incubated with the non-incorporation control D-DOPA (D-3,4-dihydroxyphenylalanine) or when incorporation was competed out by parent amino acids. The findings of the present study suggest that certain oxidized proteins could play an active role in the progression of age-related disorders by contributing to LMP (lysosomal membrane permeabilization)-initiated apoptosis and may have important implications for the long-term use of L-DOPA as a therapeutic agent in Parkinson's disease.

  • 25. Erdal, H
    et al.
    Berndtsson, M
    Castro, J
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Shoshan, M C
    Linder, S
    Induction of lysosomal membrane permeabilization by compounds that activate p53-independent apoptosis2005In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 102, no 1, p. 192-197Article in journal (Refereed)
    Abstract [en]

    The p53 protein activates cellular death programs through multiple pathways. Because the high frequency of p53 mutations in human tumors is believed to contribute to resistance to commonly used chemotherapeutic agents, it is important to identify drugs that induce p53-independent cell death and to define the mechanisms of action of such drugs. Here we screened a drug library (the National Cancer Institute mechanistic set, 879 compounds with diverse mechanisms of actions) and identified 175 compounds that induced caspase cleavage of cytokeratin-18 in cultured HCT116 colon cancer cells at ≤5 μM. Interestingly, whereas most compounds elicited a stronger apoptotic response in cells with functional p53, significant apoptosis was observed also in p53-null cells. A subset of 15 compounds showing weak or no dependence on p53 for induction of apoptosis was examined in detail. Of these compounds, 11 were capable of activating caspase-3 in enucleated cells. Seven such compounds with nonnuclear targets were found to induce lysosomal membrane permeabilization (LMP). Translocation of the lysosomal proteases cathepsin B and cathepsin D into the cytosol was observed after treatment with these drugs, and apoptosis was inhibited by pepstatin A, an inhibitor of cathepsin D. Apoptosis depended on Bax, suggesting that LMP induced a mitochondrial apoptotic pathway. We conclude that a large number of potential anticancer drugs induce p53-independent apoptosis and that LMP is a mediator of many such responses.

  • 26. Garner, B.
    et al.
    Roberg, Karin
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Qian, M.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Eaton, J W
    Truscott, RJW
    Redox availability of lens iron and copper: Implication for HO generation in cataract.1999In: Redox report, ISSN 1351-0002, E-ISSN 1743-2928, Vol. 4, p. 313-315Article in journal (Refereed)
  • 27. Guo, X-H
    et al.
    Huang, Q-B
    Chen, B
    Wang, S
    Qiang, L
    Zhu, Y
    Hou, F-F
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Zhao, M
    Advanced glycation end products induce actin rearrangement and subsequent hyperpermeability of endothelial cells2006In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 114, no 12, p. 874-883Article in journal (Refereed)
    Abstract [en]

    This study aimed to determine the effects of advanced glycation end products (AGEs) on endothelial cytoskeleton morphology and permeability, and to detect the underlying signaling mechanisms involved in these responses. Cultured endothelial cells (ECs) were exposed to AGE-modified human serum albumin (AGE-HSA), and EC cytoskeletal changes were evaluated by observing fluorescence of F-actin following ligation with labeled antibodies. Endothelial permeability was detected by measuring the flux of TRITC-albumin across the EC monolayers. To explore the signaling pathways behind AGE-induced EC alteration, ECs were treated with either soluble anti-AGE receptor (RAGE) IgG, or the MAPK inhibitors PD98059 and SB203580 before AGE-HSA administration. To further elucidate possible involvement of the ERK and p38 pathways in AGE-induced EC changes, adenovirus-carried recombinant constitutive dominant-negative forms of upstream ERK and p38 kinases, namely MEK1(A) and MKK6b(A), were pre-infected into ECs 24 h prior to AGE-HSA exposure. AGE-HSA induced actin cytoskeleton rearrangement, as well as EC hyperpermeability, in a dose and time-dependent manner. The effects were attenuated in cells pretreated with anti-RAGE IgG, PD98059 or SB203580, respectively. EC pre-infection with MEK1(A) and MKK6b(A) also alleviated the effect of AGEs. Furthermore, adenovirus-mediated administration of activated forms of either MEK1 or MKK6b alone induced rearrangement of F-actin and hyperpermeability. The results indicate that ERK and p38 MAPK play important roles in the mediation of AGE-induced EC barrier dysfunction associated with morphological changes of the F-actin. Copyright © Apmis 2006.

  • 28.
    Hydén, Dag
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Oto-Rhiono-Laryngology and Head & Neck Surgery. Östergötlands Läns Landsting, RC - Rekonstruktionscentrum, ÖNH - Öron- Näsa- Halskliniken.
    Latkovic, Stefan
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Ophthalmology. Östergötlands Läns Landsting, Reconstruction Centre, Department of Ophthalmology UHL/MH.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Laurent, Claude
    Ear involvement in ligneous conjunctivitis: A rarity or an under-diagnosed condition?2002In: Journal of Laryngology and Otology, ISSN 0022-2151, E-ISSN 1748-5460, Vol. 116, no 6, p. 482-487Article in journal (Refereed)
    Abstract [en]

    Conjunctivitis lignosa, a rare affliction of the conjunctiva, is sometimes associated with other disturbances. We present two children with concurrent conjunctivitis lignosa and ear involvement. In these two cases, there were histopathologically verified ligneous changes of the middle ears. Routine haematoxylin and eosin, van Gieson, periodic acid-Schiff (PAS) and alcian blue staining of specimens from the eyes and middle ears revealed findings typical for ligneous conjunctivitis. In addition, new histochemical and immunohistochemical studies for glycosaminoglycans on specimens from the eyes and middle ears showed that the accumulations of the amorphous, cell-deficient material stained strongly but heterogeneously for hyaluronic acid and weakly but uniformly for keratin sulphate. The staining for other glycosaminoglycans, e.g. chondroitin-4-sulphate and dermatan sulphate was confined to vessels and areas rich in collagen fibres and fibroblasts. In patients with conjunctivitis lignosa, the ear involvement may remain undiagnosed due to its resemblance to secretory otitis media with effusion. Since isolated ear involvement may occur, we advocate biopsies for routine haematoxylin and eosin, and specific staining for hyaluronic acid and keratin sulphate, also in children with protracted, refractory otitis media with atypical effusion.

  • 29.
    Karlsson, Markus
    et al.
    Linköping University, Department of Clinical and Experimental Medicine, Division of Neuroscience. Linköping University, Faculty of Health Sciences.
    Frennesson, Christina
    Linköping University, Department of Clinical and Experimental Medicine, Division of Neuroscience. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Anaesthetics, Operations and Specialty Surgery Center, Department of Ophthalmology in Linköping.
    Gustafsson, Therese
    Linköping University, Department of Clinical and Experimental Medicine, Division of Neuroscience. Linköping University, Faculty of Health Sciences.
    Brunk, Ulf
    Linköping University, Department of Medical and Health Sciences, Division of Drug Research. Linköping University, Faculty of Health Sciences.
    Erik Nilsson, Sven
    Linköping University, Department of Clinical and Experimental Medicine, Division of Neuroscience. Linköping University, Faculty of Health Sciences.
    Kurz, Tino
    Linköping University, Department of Medical and Health Sciences, Division of Drug Research. Linköping University, Faculty of Health Sciences.
    Autophagy of iron-binding proteins may contribute to the oxidative stress resistance of ARPE-19 cells2013In: Experimental Eye Research, ISSN 0014-4835, E-ISSN 1096-0007, Vol. 116, p. 359-365Article in journal (Refereed)
    Abstract [en]

    The objective of this study was to elucidate possible reasons for the remarkable resistance of human retinal pigment epithelial (RPE) cells to oxidative stress. Much oxidative damage is due to hydrogen peroxide meeting redox-active iron in the acidic and reducing lysosomal environment, resulting in the production of toxic hydroxyl radicals that may oxidize intralysosomal content, leading to lipofuscin (LF) formation or, if more extensive, to permeabilization of lysosomal membranes. Formation of LF is a risk factor for age-related macular degeneration (AMD) and known to jeopardize normal autophagic rejuvenation of vital cellular biomolecules. Lysosomal membrane permeabilization causes release of lysosomal content (redox-active iron, lytic enzymes), which may then cause cell death. Total cellular and lysosomal low-mass iron of cultured, immortalized human RPE (ARPE-19) cells was compared to that of another professional scavenger cell line, J774, using atomic absorption spectroscopy and the cytochemical sulfide-silver method (SSM). It was found that both cell lines contained comparable levels of total as well as intralysosomal iron, suggesting that the latter is mainly kept in a non-redox-active state in ARPE-19 cells. Basal levels and capacity for upregulation of the iron-binding proteins ferritin, metallothionein and heat shock protein 70 were tested in both cell lines using immunoblotting. Compared to J774 cells, ARPE-19 cells were found to contain very high basal levels of all these proteins, which could be even further upregulated following appropriate stimulation. These findings suggest that a high basal expression of iron-binding stress proteins, which during their normal autophagic turnover in lysosomes may temporarily bind iron prior to their degradation, could contribute to the unusual oxidative stress-resistance of ARPE-19 cells. A high steady state influx of such proteins into lysosomes would keep the level of lysosomal redox-active iron permanently low. This, in turn, should delay intralysosomal accumulation of LF in RPE cells, which is known to reduce autophagic turnover as well as uptake and degradation of worn out photoreceptor tips. This may explain why severe LF accumulation and AMD normally do not develop until fairly late in life, in spite of RPE cells being continuously exposed to high levels of oxygen and light, as well as large amounts of lipid-rich material.

  • 30.
    Karlsson, Markus
    et al.
    Linköping University, Department of Clinical and Experimental Medicine, Ophthalmology. Linköping University, Faculty of Health Sciences.
    Kurz, Tino
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Brunk, Ulf T.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Nilsson, Sven E.
    Linköping University, Department of Clinical and Experimental Medicine, Ophthalmology. Linköping University, Faculty of Health Sciences.
    Frennesson, Christina I.
    Linköping University, Department of Clinical and Experimental Medicine, Ophthalmology. Linköping University, Faculty of Health Sciences.
    What does the commonly used DCF test for oxidative stress really show?2010In: Biochemical Journal, ISSN 0264-6021, E-ISSN 1470-8728, Vol. 428, no 2, p. 183-90Article in journal (Refereed)
    Abstract [en]

    H(2)DCF-DA (dihydrodichlorofluorescein diacetate) is widely used to evaluate 'cellular oxidative stress'. After passing through the plasma membrane, this lipophilic and non-fluorescent compound is de-esterified to a hydrophilic alcohol [H(2)DCF (dihydrodichlorofluorescein)] that may be oxidized to fluorescent DCF (2',7'-dichlorofluorescein) by a process usually considered to involve ROS (reactive oxygen species). It is, however, not always recognized that, being a hydrophilic molecule, H(2)DCF does not cross membranes, except for the outer fenestrated mitochondrial ones. It is also not generally realized that oxidation of H(2)DCF is dependent either on Fenton-type reactions or on unspecific enzymatic oxidation by cytochrome c, for neither superoxide, nor H(2)O(2), directly oxidizes H(2)DCF. Consequently, oxidation of H(2)DCF requires the presence of either cytochrome c or of both redox-active transition metals and H(2)O(2). Redox-active metals exist mainly within lysosomes, whereas cytochrome c resides bound to the outer side of the inner mitochondrial membrane. Following exposure to H(2)DCF-DA, weak mitochondrial fluorescence was found in both the oxidation-resistant ARPE-19 cells and the much more sensitive J774 cells. This fluorescence was only marginally enhanced following short exposure to H(2)O(2), showing that by itself it is unable to oxidize H(2)DCF. Cells that were either exposed to the lysosomotropic detergent MSDH (O-methylserine dodecylamide hydrochloride), exposed to prolonged oxidative stress, or spontaneously apoptotic showed lysosomal permeabilization and strong DCF-induced fluorescence. The results suggest that DCF-dependent fluorescence largely reflects relocation to the cytosol of lysosomal iron and/or mitochondrial cytochrome c.

  • 31.
    Klionsky, Daniel J
    et al.
    Life Sciences Institute; Department of Molecular, Cellular and Developmental Biology; Department of Biological Chemistry; University of Michigan; Ann Arbor, MI, USA .
    Abdalla, Fabio C
    Laboratory of Structural and Functional Biology; Federal University of São Carlos (UFSCar); Campus Sorocaba; São Paulo State, Brazil .
    Abeliovich, Hagai
    Department of Biochemistry and Food Science; Hebrew University; Rehovot, Israel .
    Abraham, Robert T
    Acevedo-Arozena, Abraham
    Adeli, Khosrow
    Agholme, Lotta
    Linköping University, Department of Clinical and Experimental Medicine, Geriatric. Linköping University, Faculty of Health Sciences.
    Agnello, Maria
    Agostinis, Patrizia
    Aguirre-Ghiso, Julio A
    Ahn, Hyung Jun
    Ait-Mohamed, Ouardia
    Ait-Si-Ali, Slimane
    Akematsu, Takahiko
    Akira, Shizuo
    Al-Younes, Hesham M
    Al-Zeer, Munir A
    Albert, Matthew L
    Albin, Roger L
    Alegre-Abarrategui, Javier
    Aleo, Maria Francesca
    Alirezaei, Mehrdad
    Almasan, Alexandru
    Almonte-Becerril, Maylin
    Amano, Atsuo
    Amaravadi, Ravi
    Amarnath, Shoba
    Amer, Amal O
    Andrieu-Abadie, Nathalie
    Anantharam, Vellareddy
    Ann, David K
    Anoopkumar-Dukie, Shailendra
    Aoki, Hiroshi
    Apostolova, Nadezda
    Auberger, Patrick
    Baba, Misuzu
    Backues, Steven K
    Baehrecke, Eric H
    Bahr, Ben A
    Bai, Xue-Yuan
    Bailly, Yannick
    Baiocchi, Robert
    Baldini, Giulia
    Balduini, Walter
    Ballabio, Andrea
    Bamber, Bruce A
    Bampton, Edward T W
    Bánhegyi, Gábor
    Bartholomew, Clinton R
    Bassham, Diane C
    Bast, Robert C
    Batoko, Henri
    Bay, Boon-Huat
    Beau, Isabelle
    Béchet, Daniel M
    Begley, Thomas J
    Behl, Christian
    Behrends, Christian
    Bekri, Soumeya
    Bellaire, Bryan
    Bendall, Linda J
    Benetti, Luca
    Berliocchi, Laura
    Bernardi, Henri
    Bernassola, Francesca
    Besteiro, Sébastien
    Bhatia-Kissova, Ingrid
    Bi, Xiaoning
    Biard-Piechaczyk, Martine
    Blum, Janice S
    Boise, Lawrence H
    Bonaldo, Paolo
    Boone, David L
    Bornhauser, Beat C
    Bortoluci, Karina R
    Bossis, Ioannis
    Bost, Frédéric
    Bourquin, Jean-Pierre
    Boya, Patricia
    Boyer-Guittaut, Michaël
    Bozhkov, Peter V
    Brady, Nathan R
    Brancolini, Claudio
    Brech, Andreas
    Brenman, Jay E
    Brennand, Ana
    Bresnick, Emery H
    Brest, Patrick
    Bridges, Dave
    Bristol, Molly L
    Brookes, Paul S
    Brown, Eric J
    Brumell, John H
    Brunetti-Pierri, Nicola
    Brunk, Ulf T
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Bulman, Dennis E
    Bultman, Scott J
    Bultynck, Geert
    Burbulla, Lena F
    Bursch, Wilfried
    Butchar, Jonathan P
    Buzgariu, Wanda
    Bydlowski, Sergio P
    Cadwell, Ken
    Cahová, Monika
    Cai, Dongsheng
    Cai, Jiyang
    Cai, Qian
    Calabretta, Bruno
    Calvo-Garrido, Javier
    Camougrand, Nadine
    Campanella, Michelangelo
    Campos-Salinas, Jenny
    Candi, Eleonora
    Cao, Lizhi
    Caplan, Allan B
    Carding, Simon R
    Cardoso, Sandra M
    Carew, Jennifer S
    Carlin, Cathleen R
    Carmignac, Virginie
    Carneiro, Leticia A M
    Carra, Serena
    Caruso, Rosario A
    Casari, Giorgio
    Casas, Caty
    Castino, Roberta
    Cebollero, Eduardo
    Cecconi, Francesco
    Celli, Jean
    Chaachouay, Hassan
    Chae, Han-Jung
    Chai, Chee-Yin
    Chan, David C
    Chan, Edmond Y
    Chang, Raymond Chuen-Chung
    Che, Chi-Ming
    Chen, Ching-Chow
    Chen, Guang-Chao
    Chen, Guo-Qiang
    Chen, Min
    Chen, Quan
    Chen, Steve S-L
    Chen, WenLi
    Chen, Xi
    Chen, Xiangmei
    Chen, Xiequn
    Chen, Ye-Guang
    Chen, Yingyu
    Chen, Yongqiang
    Chen, Yu-Jen
    Chen, Zhixiang
    Cheng, Alan
    Cheng, Christopher H K
    Cheng, Yan
    Cheong, Heesun
    Cheong, Jae-Ho
    Cherry, Sara
    Chess-Williams, Russ
    Cheung, Zelda H
    Chevet, Eric
    Chiang, Hui-Ling
    Chiarelli, Roberto
    Chiba, Tomoki
    Chin, Lih-Shen
    Chiou, Shih-Hwa
    Chisari, Francis V
    Cho, Chi Hin
    Cho, Dong-Hyung
    Choi, Augustine M K
    Choi, DooSeok
    Choi, Kyeong Sook
    Choi, Mary E
    Chouaib, Salem
    Choubey, Divaker
    Choubey, Vinay
    Chu, Charleen T
    Chuang, Tsung-Hsien
    Chueh, Sheau-Huei
    Chun, Taehoon
    Chwae, Yong-Joon
    Chye, Mee-Len
    Ciarcia, Roberto
    Ciriolo, Maria R
    Clague, Michael J
    Clark, Robert S B
    Clarke, Peter G H
    Clarke, Robert
    Codogno, Patrice
    Coller, Hilary A
    Colombo, María I
    Comincini, Sergio
    Condello, Maria
    Condorelli, Fabrizio
    Cookson, Mark R
    Coombs, Graham H
    Coppens, Isabelle
    Corbalan, Ramon
    Cossart, Pascale
    Costelli, Paola
    Costes, Safia
    Coto-Montes, Ana
    Couve, Eduardo
    Coxon, Fraser P
    Cregg, James M
    Crespo, José L
    Cronjé, Marianne J
    Cuervo, Ana Maria
    Cullen, Joseph J
    Czaja, Mark J
    D'Amelio, Marcello
    Darfeuille-Michaud, Arlette
    Davids, Lester M
    Davies, Faith E
    De Felici, Massimo
    de Groot, John F
    de Haan, Cornelis A M
    De Martino, Luisa
    De Milito, Angelo
    De Tata, Vincenzo
    Debnath, Jayanta
    Degterev, Alexei
    Dehay, Benjamin
    Delbridge, Lea M D
    Demarchi, Francesca
    Deng, Yi Zhen
    Dengjel, Jörn
    Dent, Paul
    Denton, Donna
    Deretic, Vojo
    Desai, Shyamal D
    Devenish, Rodney J
    Di Gioacchino, Mario
    Di Paolo, Gilbert
    Di Pietro, Chiara
    Díaz-Araya, Guillermo
    Díaz-Laviada, Inés
    Diaz-Meco, Maria T
    Diaz-Nido, Javier
    Dikic, Ivan
    Dinesh-Kumar, Savithramma P
    Ding, Wen-Xing
    Distelhorst, Clark W
    Diwan, Abhinav
    Djavaheri-Mergny, Mojgan
    Dokudovskaya, Svetlana
    Dong, Zheng
    Dorsey, Frank C
    Dosenko, Victor
    Dowling, James J
    Doxsey, Stephen
    Dreux, Marlène
    Drew, Mark E
    Duan, Qiuhong
    Duchosal, Michel A
    Duff, Karen
    Dugail, Isabelle
    Durbeej, Madeleine
    Duszenko, Michael
    Edelstein, Charles L
    Edinger, Aimee L
    Egea, Gustavo
    Eichinger, Ludwig
    Eissa, N Tony
    Ekmekcioglu, Suhendan
    El-Deiry, Wafik S
    Elazar, Zvulun
    Elgendy, Mohamed
    Ellerby, Lisa M
    Eng, Kai Er
    Engelbrecht, Anna-Mart
    Engelender, Simone
    Erenpreisa, Jekaterina
    Escalante, Ricardo
    Esclatine, Audrey
    Eskelinen, Eeva-Liisa
    Espert, Lucile
    Espina, Virginia
    Fan, Huizhou
    Fan, Jia
    Fan, Qi-Wen
    Fan, Zhen
    Fang, Shengyun
    Fang, Yongqi
    Fanto, Manolis
    Fanzani, Alessandro
    Farkas, Thomas
    Farré, Jean-Claude
    Faure, Mathias
    Fechheimer, Marcus
    Feng, Carl G
    Feng, Jian
    Feng, Qili
    Feng, Youji
    Fésüs, László
    Feuer, Ralph
    Figueiredo-Pereira, Maria E
    Fimia, Gian Maria
    Fingar, Diane C
    Finkbeiner, Steven
    Finkel, Toren
    Finley, Kim D
    Fiorito, Filomena
    Fisher, Edward A
    Fisher, Paul B
    Flajolet, Marc
    Florez-McClure, Maria L
    Florio, Salvatore
    Fon, Edward A
    Fornai, Francesco
    Fortunato, Franco
    Fotedar, Rati
    Fowler, Daniel H
    Fox, Howard S
    Franco, Rodrigo
    Frankel, Lisa B
    Fransen, Marc
    Fuentes, José M
    Fueyo, Juan
    Fujii, Jun
    Fujisaki, Kozo
    Fujita, Eriko
    Fukuda, Mitsunori
    Furukawa, Ruth H
    Gaestel, Matthias
    Gailly, Philippe
    Gajewska, Malgorzata
    Galliot, Brigitte
    Galy, Vincent
    Ganesh, Subramaniam
    Ganetzky, Barry
    Ganley, Ian G
    Gao, Fen-Biao
    Gao, George F
    Gao, Jinming
    Garcia, Lorena
    Garcia-Manero, Guillermo
    Garcia-Marcos, Mikel
    Garmyn, Marjan
    Gartel, Andrei L
    Gatti, Evelina
    Gautel, Mathias
    Gawriluk, Thomas R
    Gegg, Matthew E
    Geng, Jiefei
    Germain, Marc
    Gestwicki, Jason E
    Gewirtz, David A
    Ghavami, Saeid
    Ghosh, Pradipta
    Giammarioli, Anna M
    Giatromanolaki, Alexandra N
    Gibson, Spencer B
    Gilkerson, Robert W
    Ginger, Michael L
    Goncu, Ebru
    Gongora, Céline
    Gonzalez, Claudio D
    Gonzalez, Ramon
    González-Estévez, Cristina
    González-Polo, Rosa Ana
    Gonzalez-Rey, Elena
    Gorbunov, Nikolai V
    Gorski, Sharon
    Goruppi, Sandro
    Gottlieb, Roberta A
    Gozuacik, Devrim
    Granato, Giovanna Elvira
    Grant, Gary D
    Green, Kim N
    Gregorc, Aleš
    Gros, Frédéric
    Grose, Charles
    Grunt, Thomas W
    Gual, Philippe
    Guan, Jun-Lin
    Guan, Kun-Liang
    Guichard, Sylvie M
    Gukovskaya, Anna S
    Gukovsky, Ilya
    Gunst, Jan
    Gustafsson, Asa B
    Halayko, Andrew J
    Hale, Amber N
    Halonen, Sandra K
    Hamasaki, Maho
    Han, Feng
    Han, Ting
    Hancock, Michael K
    Hansen, Malene
    Harada, Hisashi
    Harada, Masaru
    Hardt, Stefan E
    Harper, J Wade
    Harris, Adrian L
    Harris, James
    Harris, Steven D
    Hébert, Marie-Joseé
    Heidenreich, Kim A
    Helfrich, Miep H
    Helgason, Gudmundur V
    Henske, Elizabeth P
    Herman, Brian
    Herman, Paul K
    Hetz, Claudio
    Hilfiker, Sabine
    Hill, Joseph A
    Hocking, Lynne J
    Hofman, Paul
    Hofmann, Thomas G
    Höhfeld, Jörg
    Holyoake, Tessa L
    Hong, Ming-Huang
    Hood, David A
    Hotamisligil, Gökhan S
    Houwerzijl, Ewout J
    Høyer-Hansen, Maria
    Hu, Bingren
    Hu, Chien-An A
    Hu, Hong-Ming
    Hua, Ya
    Huang, Canhua
    Huang, Ju
    Huang, Shengbing
    Huang, Wei-Pang
    Huber, Tobias B
    Huh, Won-Ki
    Hung, Tai-Ho
    Hupp, Ted R
    Hur, Gang Min
    Hurley, James B
    Hussain, Sabah N A
    Hussey, Patrick J
    Hwang, Jung Jin
    Hwang, Seungmin
    Ichihara, Atsuhiro
    Ilkhanizadeh, Shirin
    Inoki, Ken
    Into, Takeshi
    Iovane, Valentina
    Iovanna, Juan L
    Ip, Nancy Y
    Isaka, Yoshitaka
    Ishida, Hiroyuki
    Isidoro, Ciro
    Isobe, Ken-ichi
    Iwasaki, Akiko
    Izquierdo, Marta
    Izumi, Yotaro
    Jaakkola, Panu M
    Jäättelä, Marja
    Jackson, George R
    Jackson, William T
    Janji, Bassam
    Jendrach, Marina
    Jeon, Ju-Hong
    Jeung, Eui-Bae
    Jiang, Hong
    Jiang, Hongchi
    Jiang, Jean X
    Jiang, Ming
    Jiang, Qing
    Jiang, Xuejun
    Jiang, Xuejun
    Jiménez, Alberto
    Jin, Meiyan
    Jin, Shengkan
    Joe, Cheol O
    Johansen, Terje
    Johnson, Daniel E
    Johnson, Gail V W
    Jones, Nicola L
    Joseph, Bertrand
    Joseph, Suresh K
    Joubert, Annie M
    Juhász, Gábor
    Juillerat-Jeanneret, Lucienne
    Jung, Chang Hwa
    Jung, Yong-Keun
    Kaarniranta, Kai
    Kaasik, Allen
    Kabuta, Tomohiro
    Kadowaki, Motoni
    Kågedal, Katarina
    Linköping University, Department of Clinical and Experimental Medicine, Experimental Pathology. Linköping University, Faculty of Health Sciences.
    Kamada, Yoshiaki
    Kaminskyy, Vitaliy O
    Kampinga, Harm H
    Kanamori, Hiromitsu
    Kang, Chanhee
    Kang, Khong Bee
    Kang, Kwang Il
    Kang, Rui
    Kang, Yoon-A
    Kanki, Tomotake
    Kanneganti, Thirumala-Devi
    Kanno, Haruo
    Kanthasamy, Anumantha G
    Kanthasamy, Arthi
    Karantza, Vassiliki
    Kaushal, Gur P
    Kaushik, Susmita
    Kawazoe, Yoshinori
    Ke, Po-Yuan
    Kehrl, John H
    Kelekar, Ameeta
    Kerkhoff, Claus
    Kessel, David H
    Khalil, Hany
    Kiel, Jan A K W
    Kiger, Amy A
    Kihara, Akio
    Kim, Deok Ryong
    Kim, Do-Hyung
    Kim, Dong-Hou
    Kim, Eun-Kyoung
    Kim, Hyung-Ryong
    Kim, Jae-Sung
    Kim, Jeong Hun
    Kim, Jin Cheon
    Kim, John K
    Kim, Peter K
    Kim, Seong Who
    Kim, Yong-Sun
    Kim, Yonghyun
    Kimchi, Adi
    Kimmelman, Alec C
    King, Jason S
    Kinsella, Timothy J
    Kirkin, Vladimir
    Kirshenbaum, Lorrie A
    Kitamoto, Katsuhiko
    Kitazato, Kaio
    Klein, Ludger
    Klimecki, Walter T
    Klucken, Jochen
    Knecht, Erwin
    Ko, Ben C B
    Koch, Jan C
    Koga, Hiroshi
    Koh, Jae-Young
    Koh, Young Ho
    Koike, Masato
    Komatsu, Masaaki
    Kominami, Eiki
    Kong, Hee Jeong
    Kong, Wei-Jia
    Korolchuk, Viktor I
    Kotake, Yaichiro
    Koukourakis, Michael I
    Kouri Flores, Juan B
    Kovács, Attila L
    Kraft, Claudine
    Krainc, Dimitri
    Krämer, Helmut
    Kretz-Remy, Carole
    Krichevsky, Anna M
    Kroemer, Guido
    Krüger, Rejko
    Krut, Oleg
    Ktistakis, Nicholas T
    Kuan, Chia-Yi
    Kucharczyk, Roza
    Kumar, Ashok
    Kumar, Raj
    Kumar, Sharad
    Kundu, Mondira
    Kung, Hsing-Jien
    Kurz, Tino
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Kwon, Ho Jeong
    La Spada, Albert R
    Lafont, Frank
    Lamark, Trond
    Landry, Jacques
    Lane, Jon D
    Lapaquette, Pierre
    Laporte, Jocelyn F
    László, Lajos
    Lavandero, Sergio
    Lavoie, Josée N
    Layfield, Robert
    Lazo, Pedro A
    Le, Weidong
    Le Cam, Laurent
    Ledbetter, Daniel J
    Lee, Alvin J X
    Lee, Byung-Wan
    Lee, Gyun Min
    Lee, Jongdae
    Lee, Ju-Hyun
    Lee, Michael
    Lee, Myung-Shik
    Lee, Sug Hyung
    Leeuwenburgh, Christiaan
    Legembre, Patrick
    Legouis, Renaud
    Lehmann, Michael
    Lei, Huan-Yao
    Lei, Qun-Ying
    Leib, David A
    Leiro, José
    Lemasters, John J
    Lemoine, Antoinette
    Lesniak, Maciej S
    Lev, Dina
    Levenson, Victor V
    Levine, Beth
    Levy, Efrat
    Li, Faqiang
    Li, Jun-Lin
    Li, Lian
    Li, Sheng
    Li, Weijie
    Li, Xue-Jun
    Li, Yan-bo
    Li, Yi-Ping
    Liang, Chengyu
    Liang, Qiangrong
    Liao, Yung-Feng
    Liberski, Pawel P
    Lieberman, Andrew
    Lim, Hyunjung J
    Lim, Kah-Leong
    Lim, Kyu
    Lin, Chiou-Feng
    Lin, Fu-Cheng
    Lin, Jian
    Lin, Jiandie D
    Lin, Kui
    Lin, Wan-Wan
    Lin, Weei-Chin
    Lin, Yi-Ling
    Linden, Rafael
    Lingor, Paul
    Lippincott-Schwartz, Jennifer
    Lisanti, Michael P
    Liton, Paloma B
    Liu, Bo
    Liu, Chun-Feng
    Liu, Kaiyu
    Liu, Leyuan
    Liu, Qiong A
    Liu, Wei
    Liu, Young-Chau
    Liu, Yule
    Lockshin, Richard A
    Lok, Chun-Nam
    Lonial, Sagar
    Loos, Benjamin
    Lopez-Berestein, Gabriel
    López-Otín, Carlos
    Lossi, Laura
    Lotze, Michael T
    Lőw, Peter
    Lu, Binfeng
    Lu, Bingwei
    Lu, Bo
    Lu, Zhen
    Luciano, Frédéric
    Lukacs, Nicholas W
    Lund, Anders H
    Lynch-Day, Melinda A
    Ma, Yong
    Macian, Fernando
    MacKeigan, Jeff P
    Macleod, Kay F
    Madeo, Frank
    Maiuri, Luigi
    Maiuri, Maria Chiara
    Malagoli, Davide
    Malicdan, May Christine V
    Malorni, Walter
    Man, Na
    Mandelkow, Eva-Maria
    Manon, Stéphen
    Manov, Irena
    Mao, Kai
    Mao, Xiang
    Mao, Zixu
    Marambaud, Philippe
    Marazziti, Daniela
    Marcel, Yves L
    Marchbank, Katie
    Marchetti, Piero
    Marciniak, Stefan J
    Marcondes, Mateus
    Mardi, Mohsen
    Marfe, Gabriella
    Mariño, Guillermo
    Markaki, Maria
    Marten, Mark R
    Martin, Seamus J
    Martinand-Mari, Camille
    Martinet, Wim
    Martinez-Vicente, Marta
    Masini, Matilde
    Matarrese, Paola
    Matsuo, Saburo
    Matteoni, Raffaele
    Mayer, Andreas
    Mazure, Nathalie M
    McConkey, David J
    McConnell, Melanie J
    McDermott, Catherine
    McDonald, Christine
    McInerney, Gerald M
    McKenna, Sharon L
    McLaughlin, BethAnn
    McLean, Pamela J
    McMaster, Christopher R
    McQuibban, G Angus
    Meijer, Alfred J
    Meisler, Miriam H
    Meléndez, Alicia
    Melia, Thomas J
    Melino, Gerry
    Mena, Maria A
    Menendez, Javier A
    Menna-Barreto, Rubem F S
    Menon, Manoj B
    Menzies, Fiona M
    Mercer, Carol A
    Merighi, Adalberto
    Merry, Diane E
    Meschini, Stefania
    Meyer, Christian G
    Meyer, Thomas F
    Miao, Chao-Yu
    Miao, Jun-Ying
    Michels, Paul A M
    Michiels, Carine
    Mijaljica, Dalibor
    Milojkovic, Ana
    Minucci, Saverio
    Miracco, Clelia
    Miranti, Cindy K
    Mitroulis, Ioannis
    Miyazawa, Keisuke
    Mizushima, Noboru
    Mograbi, Baharia
    Mohseni, Simin
    Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
    Molero, Xavier
    Mollereau, Bertrand
    Mollinedo, Faustino
    Momoi, Takashi
    Monastyrska, Iryna
    Monick, Martha M
    Monteiro, Mervyn J
    Moore, Michael N
    Mora, Rodrigo
    Moreau, Kevin
    Moreira, Paula I
    Moriyasu, Yuji
    Moscat, Jorge
    Mostowy, Serge
    Mottram, Jeremy C
    Motyl, Tomasz
    Moussa, Charbel E-H
    Müller, Sylke
    Muller, Sylviane
    Münger, Karl
    Münz, Christian
    Murphy, Leon O
    Murphy, Maureen E
    Musarò, Antonio
    Mysorekar, Indira
    Nagata, Eiichiro
    Nagata, Kazuhiro
    Nahimana, Aimable
    Nair, Usha
    Nakagawa, Toshiyuki
    Nakahira, Kiichi
    Nakano, Hiroyasu
    Nakatogawa, Hitoshi
    Nanjundan, Meera
    Naqvi, Naweed I
    Narendra, Derek P
    Narita, Masashi
    Navarro, Miguel
    Nawrocki, Steffan T
    Nazarko, Taras Y
    Nemchenko, Andriy
    Netea, Mihai G
    Neufeld, Thomas P
    Ney, Paul A
    Nezis, Ioannis P
    Nguyen, Huu Phuc
    Nie, Daotai
    Nishino, Ichizo
    Nislow, Corey
    Nixon, Ralph A
    Noda, Takeshi
    Noegel, Angelika A
    Nogalska, Anna
    Noguchi, Satoru
    Notterpek, Lucia
    Novak, Ivana
    Nozaki, Tomoyoshi
    Nukina, Nobuyuki
    Nürnberger, Thorsten
    Nyfeler, Beat
    Obara, Keisuke
    Oberley, Terry D
    Oddo, Salvatore
    Ogawa, Michinaga
    Ohashi, Toya
    Okamoto, Koji
    Oleinick, Nancy L
    Oliver, F Javier
    Olsen, Laura J
    Olsson, Stefan
    Opota, Onya
    Osborne, Timothy F
    Ostrander, Gary K
    Otsu, Kinya
    Ou, Jing-hsiung James
    Ouimet, Mireille
    Overholtzer, Michael
    Ozpolat, Bulent
    Paganetti, Paolo
    Pagnini, Ugo
    Pallet, Nicolas
    Palmer, Glen E
    Palumbo, Camilla
    Pan, Tianhong
    Panaretakis, Theocharis
    Pandey, Udai Bhan
    Papackova, Zuzana
    Papassideri, Issidora
    Paris, Irmgard
    Park, Junsoo
    Park, Ohkmae K
    Parys, Jan B
    Parzych, Katherine R
    Patschan, Susann
    Patterson, Cam
    Pattingre, Sophie
    Pawelek, John M
    Peng, Jianxin
    Perlmutter, David H
    Perrotta, Ida
    Perry, George
    Pervaiz, Shazib
    Peter, Matthias
    Peters, Godefridus J
    Petersen, Morten
    Petrovski, Goran
    Phang, James M
    Piacentini, Mauro
    Pierre, Philippe
    Pierrefite-Carle, Valérie
    Pierron, Gérard
    Pinkas-Kramarski, Ronit
    Piras, Antonio
    Piri, Natik
    Platanias, Leonidas C
    Pöggeler, Stefanie
    Poirot, Marc
    Poletti, Angelo
    Poüs, Christian
    Pozuelo-Rubio, Mercedes
    Prætorius-Ibba, Mette
    Prasad, Anil
    Prescott, Mark
    Priault, Muriel
    Produit-Zengaffinen, Nathalie
    Progulske-Fox, Ann
    Proikas-Cezanne, Tassula
    Przedborski, Serge
    Przyklenk, Karin
    Puertollano, Rosa
    Puyal, Julien
    Qian, Shu-Bing
    Qin, Liang
    Qin, Zheng-Hong
    Quaggin, Susan E
    Raben, Nina
    Rabinowich, Hannah
    Rabkin, Simon W
    Rahman, Irfan
    Rami, Abdelhaq
    Ramm, Georg
    Randall, Glenn
    Randow, Felix
    Rao, V Ashutosh
    Rathmell, Jeffrey C
    Ravikumar, Brinda
    Ray, Swapan K
    Reed, Bruce H
    Reed, John C
    Reggiori, Fulvio
    Régnier-Vigouroux, Anne
    Reichert, Andreas S
    Reiners, John J
    Reiter, Russel J
    Ren, Jun
    Revuelta, José L
    Rhodes, Christopher J
    Ritis, Konstantinos
    Rizzo, Elizete
    Robbins, Jeffrey
    Roberge, Michel
    Roca, Hernan
    Roccheri, Maria C
    Rocchi, Stephane
    Rodemann, H Peter
    Rodríguez de Córdoba, Santiago
    Rohrer, Bärbel
    Roninson, Igor B
    Rosen, Kirill
    Rost-Roszkowska, Magdalena M
    Rouis, Mustapha
    Rouschop, Kasper M A
    Rovetta, Francesca
    Rubin, Brian P
    Rubinsztein, David C
    Ruckdeschel, Klaus
    Rucker, Edmund B
    Rudich, Assaf
    Rudolf, Emil
    Ruiz-Opazo, Nelson
    Russo, Rossella
    Rusten, Tor Erik
    Ryan, Kevin M
    Ryter, Stefan W
    Sabatini, David M
    Sadoshima, Junichi
    Saha, Tapas
    Saitoh, Tatsuya
    Sakagami, Hiroshi
    Sakai, Yasuyoshi
    Salekdeh, Ghasem Hoseini
    Salomoni, Paolo
    Salvaterra, Paul M
    Salvesen, Guy
    Salvioli, Rosa
    Sanchez, Anthony M J
    Sánchez-Alcázar, José A
    Sánchez-Prieto, Ricardo
    Sandri, Marco
    Sankar, Uma
    Sansanwal, Poonam
    Santambrogio, Laura
    Saran, Shweta
    Sarkar, Sovan
    Sarwal, Minnie
    Sasakawa, Chihiro
    Sasnauskiene, Ausra
    Sass, Miklós
    Sato, Ken
    Sato, Miyuki
    Schapira, Anthony H V
    Scharl, Michael
    Schätzl, Hermann M
    Scheper, Wiep
    Schiaffino, Stefano
    Schneider, Claudio
    Schneider, Marion E
    Schneider-Stock, Regine
    Schoenlein, Patricia V
    Schorderet, Daniel F
    Schüller, Christoph
    Schwartz, Gary K
    Scorrano, Luca
    Sealy, Linda
    Seglen, Per O
    Segura-Aguilar, Juan
    Seiliez, Iban
    Seleverstov, Oleksandr
    Sell, Christian
    Seo, Jong Bok
    Separovic, Duska
    Setaluri, Vijayasaradhi
    Setoguchi, Takao
    Settembre, Carmine
    Shacka, John J
    Shanmugam, Mala
    Shapiro, Irving M
    Shaulian, Eitan
    Shaw, Reuben J
    Shelhamer, James H
    Shen, Han-Ming
    Shen, Wei-Chiang
    Sheng, Zu-Hang
    Shi, Yang
    Shibuya, Kenichi
    Shidoji, Yoshihiro
    Shieh, Jeng-Jer
    Shih, Chwen-Ming
    Shimada, Yohta
    Shimizu, Shigeomi
    Shintani, Takahiro
    Shirihai, Orian S
    Shore, Gordon C
    Sibirny, Andriy A
    Sidhu, Stan B
    Sikorska, Beata
    Silva-Zacarin, Elaine C M
    Simmons, Alison
    Simon, Anna Katharina
    Simon, Hans-Uwe
    Simone, Cristiano
    Simonsen, Anne
    Sinclair, David A
    Singh, Rajat
    Sinha, Debasish
    Sinicrope, Frank A
    Sirko, Agnieszka
    Siu, Parco M
    Sivridis, Efthimios
    Skop, Vojtech
    Skulachev, Vladimir P
    Slack, Ruth S
    Smaili, Soraya S
    Smith, Duncan R
    Soengas, Maria S
    Soldati, Thierry
    Song, Xueqin
    Sood, Anil K
    Soong, Tuck Wah
    Sotgia, Federica
    Spector, Stephen A
    Spies, Claudia D
    Springer, Wolfdieter
    Srinivasula, Srinivasa M
    Stefanis, Leonidas
    Steffan, Joan S
    Stendel, Ruediger
    Stenmark, Harald
    Stephanou, Anastasis
    Stern, Stephan T
    Sternberg, Cinthya
    Stork, Björn
    Strålfors, Peter
    Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
    Subauste, Carlos S
    Sui, Xinbing
    Sulzer, David
    Sun, Jiaren
    Sun, Shi-Yong
    Sun, Zhi-Jun
    Sung, Joseph J Y
    Suzuki, Kuninori
    Suzuki, Toshihiko
    Swanson, Michele S
    Swanton, Charles
    Sweeney, Sean T
    Sy, Lai-King
    Szabadkai, Gyorgy
    Tabas, Ira
    Taegtmeyer, Heinrich
    Tafani, Marco
    Takács-Vellai, Krisztina
    Takano, Yoshitaka
    Takegawa, Kaoru
    Takemura, Genzou
    Takeshita, Fumihiko
    Talbot, Nicholas J
    Tan, Kevin S W
    Tanaka, Keiji
    Tanaka, Kozo
    Tang, Daolin
    Tang, Dingzhong
    Tanida, Isei
    Tannous, Bakhos A
    Tavernarakis, Nektarios
    Taylor, Graham S
    Taylor, Gregory A
    Taylor, J Paul
    Terada, Lance S
    Terman, Alexei
    Tettamanti, Gianluca
    Thevissen, Karin
    Thompson, Craig B
    Thorburn, Andrew
    Thumm, Michael
    Tian, FengFeng
    Tian, Yuan
    Tocchini-Valentini, Glauco
    Tolkovsky, Aviva M
    Tomino, Yasuhiko
    Tönges, Lars
    Tooze, Sharon A
    Tournier, Cathy
    Tower, John
    Towns, Roberto
    Trajkovic, Vladimir
    Travassos, Leonardo H
    Tsai, Ting-Fen
    Tschan, Mario P
    Tsubata, Takeshi
    Tsung, Allan
    Turk, Boris
    Turner, Lorianne S
    Tyagi, Suresh C
    Uchiyama, Yasuo
    Ueno, Takashi
    Umekawa, Midori
    Umemiya-Shirafuji, Rika
    Unni, Vivek K
    Vaccaro, Maria I
    Valente, Enza Maria
    Van den Berghe, Greet
    van der Klei, Ida J
    van Doorn, Wouter
    van Dyk, Linda F
    van Egmond, Marjolein
    van Grunsven, Leo A
    Vandenabeele, Peter
    Vandenberghe, Wim P
    Vanhorebeek, Ilse
    Vaquero, Eva C
    Velasco, Guillermo
    Vellai, Tibor
    Vicencio, Jose Miguel
    Vierstra, Richard D
    Vila, Miquel
    Vindis, Cécile
    Viola, Giampietro
    Viscomi, Maria Teresa
    Voitsekhovskaja, Olga V
    von Haefen, Clarissa
    Votruba, Marcela
    Wada, Keiji
    Wade-Martins, Richard
    Walker, Cheryl L
    Walsh, Craig M
    Walter, Jochen
    Wan, Xiang-Bo
    Wang, Aimin
    Wang, Chenguang
    Wang, Dawei
    Wang, Fan
    Wang, Fen
    Wang, Guanghui
    Wang, Haichao
    Wang, Hong-Gang
    Wang, Horng-Dar
    Wang, Jin
    Wang, Ke
    Wang, Mei
    Wang, Richard C
    Wang, Xinglong
    Wang, Xuejun
    Wang, Ying-Jan
    Wang, Yipeng
    Wang, Zhen
    Wang, Zhigang Charles
    Wang, Zhinong
    Wansink, Derick G
    Ward, Diane M
    Watada, Hirotaka
    Waters, Sarah L
    Webster, Paul
    Wei, Lixin
    Weihl, Conrad C
    Weiss, William A
    Welford, Scott M
    Wen, Long-Ping
    Whitehouse, Caroline A
    Whitton, J Lindsay
    Whitworth, Alexander J
    Wileman, Tom
    Wiley, John W
    Wilkinson, Simon
    Willbold, Dieter
    Williams, Roger L
    Williamson, Peter R
    Wouters, Bradly G
    Wu, Chenghan
    Wu, Dao-Cheng
    Wu, William K K
    Wyttenbach, Andreas
    Xavier, Ramnik J
    Xi, Zhijun
    Xia, Pu
    Xiao, Gengfu
    Xie, Zhiping
    Xie, Zhonglin
    Xu, Da-zhi
    Xu, Jianzhen
    Xu, Liang
    Xu, Xiaolei
    Yamamoto, Ai
    Yamamoto, Akitsugu
    Yamashina, Shunhei
    Yamashita, Michiaki
    Yan, Xianghua
    Yanagida, Mitsuhiro
    Yang, Dun-Sheng
    Yang, Elizabeth
    Yang, Jin-Ming
    Yang, Shi Yu
    Yang, Wannian
    Yang, Wei Yuan
    Yang, Zhifen
    Yao, Meng-Chao
    Yao, Tso-Pang
    Yeganeh, Behzad
    Yen, Wei-Lien
    Yin, Jia-jing
    Yin, Xiao-Ming
    Yoo, Ook-Joon
    Yoon, Gyesoon
    Yoon, Seung-Yong
    Yorimitsu, Tomohiro
    Yoshikawa, Yuko
    Yoshimori, Tamotsu
    Yoshimoto, Kohki
    You, Ho Jin
    Youle, Richard J
    Younes, Anas
    Yu, Li
    Yu, Long
    Yu, Seong-Woon
    Yu, Wai Haung
    Yuan, Zhi-Min
    Yue, Zhenyu
    Yun, Cheol-Heui
    Yuzaki, Michisuke
    Zabirnyk, Olga
    Silva-Zacarin, Elaine
    Zacks, David
    Zacksenhaus, Eldad
    Zaffaroni, Nadia
    Zakeri, Zahra
    Zeh, Herbert J
    Zeitlin, Scott O
    Zhang, Hong
    Zhang, Hui-Ling
    Zhang, Jianhua
    Zhang, Jing-Pu
    Zhang, Lin
    Zhang, Long
    Zhang, Ming-Yong
    Zhang, Xu Dong
    Zhao, Mantong
    Zhao, Yi-Fang
    Zhao, Ying
    Zhao, Zhizhuang J
    Zheng, Xiaoxiang
    Zhivotovsky, Boris
    Zhong, Qing
    Zhou, Cong-Zhao
    Zhu, Changlian
    Zhu, Wei-Guo
    Zhu, Xiao-Feng
    Zhu, Xiongwei
    Zhu, Yuangang
    Zoladek, Teresa
    Zong, Wei-Xing
    Zorzano, Antonio
    Zschocke, Jürgen
    Zuckerbraun, Brian
    Guidelines for the use and interpretation of assays for monitoring autophagy2012In: Autophagy, ISSN 1554-8627, Vol. 8, no 4, p. 445-544Article, review/survey (Refereed)
    Abstract [en]

    In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.

  • 32.
    Klionsky, Daniel J.
    et al.
    University of Michigan, USA.
    Abeliovich, Hagai
    Hebrew University of Jerusalem, Israel.
    Agostinis, Patrizia
    Catholic University of Louvain, Belgium.
    Agrawal, Devendra K.
    Creighton University, USA.
    Aliev, Giumrakch
    University of Texas San Antonio, TX USA.
    S. Askew, David
    University of Cincinnati, USA.
    Baba, Misuzu
    Japan Womens University, Japan.
    H. Baehrecke, Eric
    University of Massachusetts, MA USA.
    A. Bahr, Ben
    University of Connecticut, CT USA.
    Ballabio, Andrea
    Telethon Institute Genet and Med, Italy.
    A. Bamber, Bruce
    University of Toledo, OH 43606 USA .
    C. Bassham, Diane
    Iowa State University, IA USA Iowa State University, IA USA .
    Bergamini, Ettore
    University of Pisa, Italy .
    Bi, Xiaoning
    Western University of Health Science, CA USA .
    Biard-Piechaczyk, Martine
    UM2, France .
    S. Blum, Janice
    Indiana University, IN 46202 USA .
    E. Breclesen, Dale
    Bucks Institute Age Research, CA USA .
    L. Brodsky, Jeffrey
    University of Pittsburgh, PA 15260 USA Massachusetts Gen Hospital, MA USA .
    H. Brumell, John
    Hospital Sick Children, Canada .
    Brunk, Ulf T.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Bursch, Wilfried
    Medical University of Vienna, Austria .
    Camougrand, Nadine
    University of Bordeaux 2, France .
    Cebollero, Eduardo
    CSIC, Spain .
    Cecconi, Francesco
    University of Roma Tor Vergata, Italy University of Roma Tor Vergata, Italy .
    Chen, Yingyu
    Peking University, Peoples R China .
    Chin, Lih-Shen
    Emory University School of Medicine.
    Choi, Augustine
    Emory University, GA 30322 USA .
    T. Chu, Charleen
    Harvard University, MA USA.
    Chung, Jongkyeong
    University of Pittsburgh, PA USA Korea Adv Institute Science and Technology, South Korea .
    G. H. Clarke, Peter
    University of Lausanne, Switzerland .
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    Safar Centre Resuscitat Research, PA USA .
    G. Clarke, Steven
    University of Calif Los Angeles, CA 90024 USA University of Calif Los Angeles, CA 90024 USA .
    Clave, Corinne
    University of Bordeaux 2, France .
    L. Cleveland, John
    Scripps Research Institute, FL USA .
    Codogno, Patrice
    University of Paris 11, France INSERM, France .
    I. Colombo, Maria
    University of Nacl Cuyo, Argentina .
    Coto-Montes, Ana
    University of Oviedo, Spain .
    M. Cregg, James
    Keck Grad Institute Appl Science, CA USA .
    Maria Cuervo, Ana
    Albert Einstein Coll Med, NY 10467 USA .
    Debnath, Jayanta
    University of Calif San Francisco, CA USA .
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    Lab Nazl Consorzio Interuniv Biotecnol, Italy University of Cincinnati, OH USA .
    B. Dennis, Patrick
    USN Medical Oncol, MD 20892 USA .
    A. Dennis, Phillip
    USN Medical Oncol, MD 20892 USA .
    Deretic, Vojo
    University of New Mexico, NM 87131 USA .
    J. Devenish, Rodney
    Monash University, Australia Monash University, Australia .
    Di Sano, Federica
    University of Roma Tor Vergata, Italy .
    Fred Dice, J.
    Tufts University, MA 02111 USA .
    DiFiglia, Marian
    Massachusetts Gen Hospital, MA USA .
    Dinesh-Kumar, Savithramma
    Yale University, CT USA .
    W. Distelhorst, Clark
    Case Western Reserve University, OH 44106 USA Case Western Reserve University, OH 44106 USA University Hospital Cleveland, OH 44106 USA Case Western Reserve University, OH 44106 USA .
    Djavaheri-Mergny, Mojgan
    University of Paris 11, France INSERM, France .
    C. Dorsey, Frank
    The Scripps Research Institute.
    Droege, Wulf
    Immunotec Research Ltd, Canada .
    Dron, Michel
    INRA, France .
    A. Jr. Dunn, William
    University of Florida, FL USA .
    Duszenko, Michael
    University of Tubingen, Germany .
    Tony Eissa, N.
    Baylor University, TX 77030 USA .
    Elazar, Zvulun
    Weizmann Institute Science, Israel .
    Esclatine, Audrey
    University of Paris 11, France INSERM, France .
    Eskelinen, Eeva-Liisa
    University of Helsinki, Finland .
    Fesues, Laszlo
    University of Debrecen, Hungary University of Debrecen, Hungary .
    D. Finley, Kim
    Salk Institute Biol Studies, CA USA .
    M. Fuentes, Jose
    University of Extremadura, Spain .
    Fueyo, Juan
    University of Texas Houston, TX 77030 USA .
    Fujisaki, Kozo
    Kagoshima University, Japan .
    Galliot, Brigitte
    University of Geneva, Switzerland .
    Gao, Fen-Biao
    University of Calif San Francisco, CA 94143 USA University of Calif San Francisco, CA 94143 USA .
    A. Gewirtz, David
    Virginia Commonwealth University, VA USA Virginia Commonwealth University, VA USA .
    B. Gibson, Spencer
    Manitoba Institute Cell Biol, Canada .
    Gohla, Antje
    University of Dusseldorf, Germany .
    L. Goldberg, Alfred
    Harvard University, MA USA .
    Gonzalez, Ramon
    CSIC, Spain .
    Gonzalez-Estevez, Cristina
    University of Nottingham, England .
    Gorski, Sharon
    British Columbia Cancer Agency, Canada .
    A. Gottlieb, Roberta
    San Diego State University, CA 92182 USA .
    Haussinger, Dieter
    University of Dusseldorf, Germany .
    He, You-Wen
    Duke University, NC USA .
    Heidenreich, Kim
    University of Colorado, CO USA .
    A. Hill, Joseph
    University of Texas SW Medical Centre Dallas, TX 75390 USA .
    Hoyer-Hansen, Maria
    Danish Cancer Soc, Denmark Danish Cancer Soc, Denmark .
    Hu, Xun
    Zhejiang University, Peoples R China .
    Huang, Wei-Pang
    National Taiwan University, Taiwan .
    Iwasaki, Akiko
    Yale University, CT USA .
    Jaattela, Marja
    University of Debrecen, Hungary University of Debrecen, Hungary .
    T. Jackson, William
    Medical Coll Wisconsin, WI 53226 USA .
    Jiang, Xuejun
    Mem Sloan Kettering Cancer Centre, NY 10021 USA .
    Jin, Shengkan
    University of Medical and Dent New Jersey, NJ 08854 USA .
    Johansen, Terje
    University of Tromso, Norway .
    U. Jung, Jae
    University of So Calif, CA USA .
    Kadowaki, Motoni
    Niigata University, Japan .
    Kang, Chanhee
    University of Texas SW Medical Centre Dallas, TX 75390 USA .
    Kelekar, Ameeta
    University of Minnesota, MN USA .
    H. Kessel, David
    Wayne State University, MI USA .
    A. K. W. Kiel, Jan
    University of Groningen, Netherlands .
    Pyo Kim, Hong
    University of Pittsburgh, PA USA .
    Kimchi, Adi
    Weizmann Institute Science, Israel .
    J. Kinsella, Timothy
    University Hospital Cleveland, OH 44106 USA .
    Kiselyov, Kirill
    University of Pittsburgh, PA 15260 USA .
    Kitamoto, Katsuhiko
    University of Tokyo, Japan .
    Knecht, Erwin
    Centre Invest Principe Felipe, Spain .
    Komatsu, Masaaki
    Tokyo Metropolitan Institute Medical Science, Japan .
    Kominami, Eiki
    Juntendo University, Japan .
    Kondo, Seiji
    University of Texas MD Anderson Cancer Center.
    L. Kovacs, Attila
    University of Texas MD Anderson Cancer Centre, TX USA .
    Kroemer, Guido
    Eotvos Lorand University, Hungary Institute Gustave Roussy, France University of Paris 11, France .
    Kuan, Chia-Yi
    Cincinnati Childrens Hospital Research Fdn, OH USA .
    Kumar, Rakesh
    University of Penn, PA 19104 USA .
    Kundu, Mondira
    University of Laval, Canada .
    Landry, Jacques
    Eastern Michigan University, MI 48197 USA .
    Laporte, Marianne
    Eastern Michigan University.
    Le, Weidong
    Shanghai Jiao Tong University, Peoples R China Chinese Academic Science, Peoples R China .
    Lei, Huan-Yao
    National Cheng Kung University, Taiwan .
    J. Lenardo, Michael
    NIAID, MD USA .
    Levine, Beth
    University of Texas SW Medical Centre Dallas, TX 75390 USA University of Texas SW Medical Centre Dallas, TX 75390 USA .
    Lieberman, Andrew
    University of Michigan, MI USA .
    Lim, Kah-Leong
    National Institute Neurosci, Singapore .
    Lin, Fu-Cheng
    Zhejiang University, Peoples R China .
    Liou, Willisa
    Chang Gung University.
    F. Liu, Leroy
    University of Medical and Dent New Jersey, NJ 08854 USA National Research Centre Environm and Heatlh, Germany .
    Lopez-Berestein, Gabriel
    University of Texas MD Anderson Cancer Centre, TX USA .
    Lopez-Otin, Carlos
    University of Oviedo, Spain .
    Lu, Bo
    Vanderbilt University, TN USA .
    F. Macleod, Kay
    University of Chicago, IL 60637 USA Ist Super Sanita, Italy .
    Malorni, Walter
    Istituto Superiore di Sanita.
    Martinet, Wim
    University of Antwerp, Belgium .
    Matsuoka, Ken
    Kyushu University, Japan .
    Mautner, Josef
    GSF-National Research Center for Environment and Health.
    J. Meijer, Alfred
    University of Amsterdam, Netherlands .
    Melendez, Alicia
    CUNY, NY USA .
    Michels, Paul
    Catholic University of Louvain, Belgium Catholic University of Louvain, Belgium .
    Miotto, Giovanni
    University of Padua, Italy .
    P. Mistiaen, Wilhelm
    University of Coll Antwerp, Belgium .
    Mizushima, Noboru
    Tokyo Medical and Dent University, Japan .
    Mograbi, Baharia
    INSERM, France IFR 50, France .
    Monastyrska, Iryna
    University of Utrecht, Netherlands .
    N. Moore, Michael
    Plymouth Marine Lab, England .
    I. Moreira, Paula
    Centre Neurosci and Cell Biol, Portugal .
    Moriyasu, Yuji
    Saitama University, Japan .
    Motyl, Tomasz
    Agriculture University of Warsaw, Poland .
    Muenz, Christian
    Rockefeller University, NY 10021 USA .
    O. Murphy, Leon
    Novartis Institute Biomed Research, MA USA .
    I. Naqvi, Naweed
    National University of Singapore, Singapore .
    Neufeld, Thomas
    University of Minnesota.
    Nishino, Ichizo
    National Centre Neurol and Psychiat, Japan .
    A. Nixon, Ralph
    NYU, NY USA .
    Noda, Takeshi
    Osaka University, Japan .
    Nuernberg, Bernd
    University of Dusseldorf, Germany .
    Ogawa, Michinaga
    University of Tokyo, Japan .
    L. Oleinick, Nancy
    Case Western Reserve University, OH USA Case Western Reserve University, OH 44106 USA Case Western Reserve University, OH 44106 USA Case Western Reserve University, OH 44106 USA .
    J. Olsen, Laura
    University of Michigan, MI 48109 USA .
    Ozpolat, Bulent
    University of Texas MD Anderson Cancer Centre, TX USA .
    Paglin, Shoshana
    Chaim Sheba Medical Centre, Israel .
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    Louisiana State University, LA USA .
    Papassideri, Issidora
    Department Cell Biol and Biophys, Greece .
    Parkes, Miles
    University of Cambridge, England .
    H. Perlmutter, David
    University of Pittsburgh, PA 15261 USA Childrens Hospital Pittsburgh, PA 15213 USA .
    Perry, George
    University of Texas San Antonio, TX USA .
    Piacentini, Mauro
    University of Roma Tor Vergata, Italy .
    Pinkas-Kramarski, Ronit
    Tel Aviv University, Israel .
    Prescott, Mark
    Monash University, Australia .
    Proikas-Cezanne, Tassula
    University of Tubingen, Germany .
    Raben, Nina
    NIAMSD, MD USA .
    Rami, Abdelhaq
    Clin JWG University, Germany .
    Reggiori, Fulvio
    University of Utrecht, Netherlands .
    Rohrer, Baerbel
    Medical University of S Carolina, SC 29425 USA .
    C. Rubinsztein, David
    Cambridge Institute Medical Research, England .
    M. Ryan, Kevin
    Beatson Institute Cancer Research, Scotland .
    Sadoshima, Junichi
    University of Medical and Dent New Jersey, NJ 07103 USA .
    Sakagami, Hiroshi
    Meikai University, Japan .
    Sakai, Yasuyoshi
    Kyoto University, Japan JST, Japan .
    Sandri, Marco
    University of Padua, Italy Venetan Institute Molecular Med, Italy .
    Sasakawa, Chihiro
    University of Tokyo, Japan .
    Sass, Miklos
    University of Oslo, Norway .
    Schneider, Claudio
    Laboratorio Nazionale Consorzio Interuniversitario Biotecnologie.
    O. Seglen, Per
    University of Wyoming, WY 82071 USA .
    Seleverstov, Oleksandr
    University of Oslo, Norway .
    Settleman, Jeffre
    Massachusetts General Hospital Cancer Center.
    J. Shacka, John
    University of Alabama Birmingham, AL 35294 USA .
    M. Shapiro, Irving
    Thomas Jefferson University, PA 19107 USA .
    Sibirny, Andrei
    National Academic Science Ukraine, Ukraine .
    C. M. Silva-Zacarin, Elaine
    University of Federal Sao Carlos, Brazil .
    Simon, Hans-Uwe
    University of Bern, Switzerland .
    Simone, Cristiano
    Ist Ric Farmacol Mario Negri, Italy .
    Simonsen, Anne
    University of Oslo, Norway Norwegian Radium Hospital, Norway .
    A. Smith, Mark
    Case Western Reserve University, OH 44106 USA .
    Spanel-Borowski, Katharina
    University of Leipzig, Germany .
    Srinivas, Vickram
    Thomas Jefferson University, PA 19107 USA .
    Steeves, Meredith
    Scripps Research Institute, FL USA .
    Stenmark, Harald
    Norwegian Radium Hospital, Norway .
    E. Stromhaug, Per
    University of Missouri, MO USA .
    S. Subauste, Carlos
    Case Western Reserve University, OH USA Case Western Reserve University, OH USA .
    Sugimoto, Seiichiro
    National Hospital Org, Japan .
    Sulzer, David
    Columbia University, NY USA Columbia University, NY USA .
    Suzuki, Toshihiko
    University of Ryukyus, Japan .
    S. Swanson, Michele
    University of Michigan, MI 48109 USA .
    Takeshita, Fumihiko
    Yokohama City University, Japan .
    J. Talbot, Nicholas.
    University of Exeter, England .
    Talloczy, Zsolt
    Columbia University, NY USA Columbia University, NY USA .
    Tanaka, Keiji
    Tokyo Metropolitan Institute Medical Science, Japan Tohoku University, Japan .
    Tanaka, Kozo
    Tokyo Metropolitan Institute Medical Science, Japan Tohoku University, Japan .
    Tanida, Isei
    National Institute Infect Disease, Japan .
    S. Taylor, Graham
    University of Birmingham, England .
    Paul Taylor, J.
    University of Penn, PA 19104 USA .
    Terman, Alexei
    Linköping University, Department of Clinical and Experimental Medicine, Geriatric. Linköping University, Faculty of Health Sciences.
    Tettamanti, Gianluca
    University of Insubria, Italy .
    B. Thompson, Craig
    University of Penn, PA 19104 USA .
    Thumm, Michael
    University of Gottingen, Germany .
    M. Tolkovsky, Aviva
    University of Cambridge, England .
    A. Tooze, Sharon
    Cancer Research UK London Research Institute, England .
    Truant, Ray
    McMaster University, Canada .
    V. Tumanovska, Lesya
    AA Bogomolets Physiol Institute, Ukraine .
    Uchiyama, Yasuo
    Osaka University, Japan .
    Ueno, Takashi
    Juntendo University, Japan .
    L. Uzcategui, Nestor
    Central University of Venezuela, Venezuela .
    van der Klei, Ida
    University of Groningen, Netherlands .
    C. Vaquero, Eva
    Hospital Clin Barcelona, Spain .
    Vellai, Tibor
    Eotvos Lorand University, Hungary .
    W. Vogel, Michael
    Maryland Psychiat Research Centre, MD 21228 USA .
    Wang, Hong-Gang
    H Lee Moffitt Cancer Centre and Research Institute, FL USA .
    Webster, Paul
    House Ear Research Institute, CA USA .
    W. Wiley, John
    University of Michigan, MI 48109 USA .
    Xi, Zhijun
    Peking University, Peoples R China .
    Xiao, Gutian
    University of Pittsburgh, PA USA .
    Yahalom, Joachim
    Memorial Sloan-Kettering Cancer Center.
    Yang, Jin-Ming
    University of Medical and Dent New Jersey, NJ USA .
    Yap, George
    University of Medical and Dent New Jersey, NJ 07103 USA .
    Yin, Xiao-Min
    University of Pittsburgh, PA USA .
    Yoshimori, Tamotsu
    Osaka University, Japan .
    Yu, Li
    NIAID, MD USA .
    Yue, Zhenyu
    Mt Sinai School Med, NY USA .
    Yuzaki, Michisuke
    Keio University, Japan .
    Zabirnyk, Olga
    NCI, MD USA National Institute Heatlh, MD USA .
    Zheng, Xiaoxiang
    Zhejiang University, Peoples R China .
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    Case Western Reserve University, OH 44106 USA .
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    Baylor University, TX 77030 USA .
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    Columbia University, NY USA .
    Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes2008In: Autophagy, ISSN 1554-8627, E-ISSN 1554-8635, Vol. 4, no 2, p. 151-175Article, review/survey (Refereed)
    Abstract [en]

    Research in autophagy continues to accelerate,1 and as a result many new scientists are entering the field. Accordingly, it is important to establish a standard set of criteria for monitoring macroautophagy in different organisms. Recent reviews have described the range of assays that have been used for this purpose.2,3 There are many useful and convenient methods that can be used to monitor macroautophagy in yeast, but relatively few in other model systems, and there is much confusion regarding acceptable methods to measure macroautophagy in higher eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers of autophagosomes versus those that measure flux through the autophagy pathway; thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from fully functional autophagy that includes delivery to, and degradation within, lysosomes (in most higher eukaryotes) or the vacuole (in plants and fungi). Here, we present a set of guidelines for the selection and interpretation of the methods that can be used by investigators who are attempting to examine macroautophagy and related processes, as well as by reviewers who need to provide realistic and reasonable critiques of papers that investigate these processes. This set of guidelines is not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to verify an autophagic response.

  • 33.
    Kurz, Tino
    et al.
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Brunk , Ulf
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Autophagy of HSP70 and chelation of lysosomal iron in a non-redox-active form2009In: AUTOPHAGY, ISSN 1554-8627 , Vol. 5, no 1, p. 93-95Article in journal (Refereed)
    Abstract [en]

    Lysosomes contain most of the cells supply of labile iron, which makes them sensitive to oxidative stress. To keep lysosomal labile iron at a minimum, a cellular strategy might be to autophagocytose iron-binding proteins that temporarily would chelate iron in a nonredox-active form. Previously we have shown that autophagy of metallothioneins, as well as of non-Fe-saturated ferritin, meets this goal. Here we add another stress-regulated protein to the list, namely HSP70.

  • 34.
    Kurz, Tino
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Brunk, Ulf
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Terman, Alexei
    Department of Clinical Pathology and Cytology, Karolinska University Hospital in Huddinge, .
    Oxidative Stress and Lysosomes2011In: Principles of Free Radical Biomedicine / [ed] Editor: K. Pantopoulos and H. Schipper, pp., Nova Science Publishers, Inc., 2011, p. 1-18Chapter in book (Refereed)
    Abstract [en]

    Recent years have witnessed an avalanche of new knowledge implicating free radicals in virtually every aspect of biology and medicine. It is now axiomatic that the regulated accumulation of reactive oxygen species (ROS) contributes to organismal health and well-being and that ROS serve as signaling molecules involved in cell growth, differentiation, gene regulation, replicative senescence and apoptosis. This book is an interdisciplinary text broken up into three consecutive volumes on the biochemistry and cellular/molecular biology of free radicals, transition metals, oxidants and antioxidants, and the role of oxidative stress in human health and disease.

  • 35.
    Kurz, Tino
    et al.
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Eaton, John W.
    University of Louisville.
    Brunk, Ulf
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Redox Activity Within the Lysosomal Compartment: Implications for Aging and Apoptosis2010In: Antioxidants and Redox Signaling, ISSN 1523-0864, E-ISSN 1557-7716, Vol. 13, no 4, p. 511-523Article, review/survey (Refereed)
    Abstract [en]

    The lysosome is a redox-active compartment containing low-mass iron and copper liberated by autophagic degradation of metalloproteins. The acidic milieu and high concentration of thiols within lysosomes will keep iron in a reduced ( ferrous) state, which can react with endogenous or exogenous hydrogen peroxide. Consequent intralysosomal Fenton reactions may give rise to the formation of lipofuscin or "age pigment that accumulates in long-lived postmitotic cells that cannot dilute it by division. Extensive accumulation of lipofuscin seems to hinder normal autophagy and may be an important factor behind aging and age-related pathologies. Enhanced oxidative stress causes lysosomal membrane permeabilization, with ensuing relocation to the cytosol of iron and lysosomal hydrolytic enzymes, with resulting apoptosis or necrosis. Lysosomal copper is normally not redox active because it will form non-redox-active complexes with various thiols. However, if cells are exposed to lysosomotropic chelators that do not bind all the copper coordinates, highly redox-active complexes may form, with ensuing extensive lysosomal Fenton-type reactions and loss of lysosomal stability. Because many malignancies seem to have increased amounts of copper-containing macromolecules that are turned over by autophagy, it is conceivable that lysosomotropic copper chelators may be used in the future in ROS-based anticancer therapies.

  • 36.
    Kurz, Tino
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Eaton, John W.
    University of Louisville.
    Brunk, Ulf
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    The role of lysosomes in iron metabolism and recycling2011In: International Journal of Biochemistry and Cell Biology, ISSN 1357-2725, E-ISSN 1878-5875, Vol. 43, no 12, p. 1686-1697Article, review/survey (Refereed)
    Abstract [en]

    Iron is the most abundant transition metal in the earths crust. It cycles easily between ferric (oxidized; Fe(III)) and ferrous (reduced; Fe(II)) and readily forms complexes with oxygen, making this metal a central player in respiration and related redox processes. However, loose iron, not within heme or iron-sulfur cluster proteins, can be destructively redox-active, causing damage to almost all cellular components, killing both cells and organisms. This may explain why iron is so carefully handled by aerobic organisms. Iron uptake from the environment is carefully limited and carried out by specialized iron transport mechanisms. One reason that iron uptake is tightly controlled is that most organisms and cells cannot efficiently excrete excess iron. When even small amounts of intracellular free iron occur, most of it is safely stored in a non-redox-active form in ferritins. Within nucleated cells, iron is constantly being recycled from aged iron-rich organelles such as mitochondria and used for construction of new organelles. Much of this recycling occurs within the lysosome, an acidic digestive organelle. Because of this, most lysosomes contain relatively large amounts of redox-active iron and are therefore unusually susceptible to oxidant-mediated destabilization or rupture. In many cell types, iron transit through the lysosomal compartment can be remarkably brisk. However, conditions adversely affecting lysosomal iron handling (or oxidant stress) can contribute to a variety of acute and chronic diseases. These considerations make normal and abnormal lysosomal handling of iron central to the understanding and, perhaps, therapy of a wide range of diseases.

  • 37.
    Kurz, Tino
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Gustafsson, Bertil
    Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre for Diagnostics, Department of Clinical Pathology and Clinical Genetics. Linköping University, Department of Clinical and Experimental Medicine, Experimental Pathology.
    Brunk, Ulf
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Cell sensitivity to oxidative stress is influenced by ferritin autophagy2011In: FREE RADICAL BIOLOGY AND MEDICINE, ISSN 0891-5849, Vol. 50, no 11, p. 1647-1658Article in journal (Refereed)
    Abstract [en]

    To test the consequences of lysosomal degradation of differently iron-loaded ferritin molecules and to mimic ferritin autophagy under iron-overload and normal conditions, J774 cells were allowed to endocytose heavily iron loaded ferritin, probably with some adventitious iron (Fe-Ft), or iron-free apo-ferritin (apo-Ft). When cells subsequently were exposed to a bolus dose of hydrogen peroxide, apo-Ft prevented lysosomal membrane permeabilization (LMP), whereas Fe-Ft enhanced LMP. A 4-h pulse of Fe-Ft initially increased oxidative stress-mediated LMP that was reversed after another 3 h under standard culture conditions, suggesting that lysosomal iron is rapidly exported from lysosomes, with resulting upregulation of apo-ferritin that supposedly is autophagocytosed, thereby preventing LMP by binding intralysosomal redox-active iron. The obtained data suggest that upregulation of the stress protein ferritin is a rapid adaptive mechanism that counteracts LMP and ensuing apoptosis during oxidative stress. In addition, prolonged iron starvation was found to induce apoptotic cell death that, interestingly, was preceded by LMP, suggesting that LMP is a more general phenomenon in apoptosis than so far recognized. The findings provide new insights into aging and neurodegenerative diseases that are associated with enhanced amounts of cellular iron and show that lysosomal iron loading sensitizes to oxidative stress.

  • 38.
    Kurz, Tino
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Gustafsson, Bertil
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Intralysosomal iron chelation protects against oxidative stress-induced cellular damage2006In: The FEBS Journal, ISSN 1742-464X, E-ISSN 1742-4658, Vol. 273, no 13, p. 3106-3117Article in journal (Refereed)
    Abstract [en]

    Oxidant-induced cell damage may be initiated by peroxidative injury to lysosomal membranes, catalyzed by intralysosomal low mass iron that appears to comprise a major part of cellular redox-active iron. Resulting relocation of lytic enzymes and low mass iron would result in secondary harm to various cellular constituents. In an effort to further clarify this still controversial issue, we tested the protective effects of two potent iron chelators - the hydrophilic desferrioxamine (dfo) and the lipophilic salicylaldehyde isonicotinoyl hydrazone (sih), using cultured lysosome-rich macrophage-like J774 cells as targets. dfo slowly enters cells via endocytosis, while the lipophilic sih rapidly distributes throughout the cell. Following dfo treatment, long-term survival of cells cannot be investigated because dfo by itself, by remaining inside the lysosomal compartment, induces apoptosis that probably is due to iron starvation, while sih has no lasting toxic effects if the exposure time is limited. Following preincubation with 1 mm dfo for 3 h or 10 μm sih for a few minutes, both agents provided strong protection against an ensuing ∼LD50 oxidant challenge by preventing lysosomal rupture, ensuing loss of mitochondrial membrane potential, and apoptotic/necrotic cell death. It appears that once significant lysosomal rupture has occurred, the cell is irreversibly committed to death. The results lend strength to the concept that lysosomal membranes, normally exposed to redox-active iron in high concentrations, are initial targets of oxidant damage and support the idea that chelators selectively targeted to the lysosomal compartment may have therapeutic utility in diminishing oxidant-mediated cell injury. © 2006 The Authors.

  • 39.
    Kurz, Tino
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Karlsson, Markus
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences.
    Brunk, Ulf
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Erik Nilsson, Sven
    Linköping University, Department of Clinical and Experimental Medicine, Ophthalmology. Linköping University, Faculty of Health Sciences.
    Frennesson, Christina
    Linköping University, Department of Clinical and Experimental Medicine, Ophthalmology. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of Ophthalmology UHL/MH.
    ARPE-19 retinal pigment epithelial cells are highly resistant to oxidative stress and exercise strict control over their lysosomal redox-active iron2009In: AUTOPHAGY, ISSN 1554-8627, Vol. 5, no 4, p. 494-501Article in journal (Refereed)
    Abstract [en]

    Normal retinal pigment epithelial (RPE) cells are postmitotic, long-lived and basically not replaced. Daily, they phagocytose substantial amounts of lipid-rich material (photoreceptor outer segment discs), and they do so in the most oxygenated part of the body-the retina. One would imagine that this state of affairs should be associated with a rapid formation of the age pigment lipofuscin (LF). However, LF accumulation is slow and reaches significant amounts only late in life when, if substantial, it often coincides with or causes age-related macular degeneration. LF formation occurs inside the lysosomal compartment as a result of iron-catalyzed peroxidation and polymerization. This process requires phagocytosed or autophagocytosed material under degradation, but also the presence of redox-active low mass iron and hydrogen peroxide. To gain some information on how RPE cells are able to evade LF formation, we investigated the response of immortalized human RPE cells (ARPE-19) to oxidative stress with/without the protection of a strong iron-chelator. The cells were found to be extremely resistant to hydrogen peroxide-induced lysosomal rupture and ensuing cell death. This marked resistance to oxidative stress was not explained by enhanced degradation of hydrogen peroxide, but to a certain extent further increased by the potent lipophilic iron chelator STH. The cells were also able to survive, and even replicate, at high concentrations of SIH and showed a high degree of basal autophagic flux. We hypothesize that RPE cells have a highly developed capacity to keep lysosomal iron in a nonredox-active form, perhaps by pronounced autophagy of iron-binding proteins in combination with an ability to rapidly relocate low mass iron from the lysosomal compartment.

  • 40.
    Kurz, Tino
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    Leake, A
    von Zglinicki, T
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    Lysosomal redox-active iron is important for oxidative stress-induced DNA damage2003In: Free radical research, ISSN 1071-5762, E-ISSN 1029-2470, Vol. 37, p. 107-107Conference paper (Other academic)
  • 41.
    Kurz, Tino
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Leake, Alan
    von Zglinicki, Thomas
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Lysosomal redox-active iron is important for oxidative stress-induced DNA damage2004In: Annals of the New York Academy of Sciences, ISSN 0077-8923, E-ISSN 1749-6632, Vol. 1019, p. 285-288Article in journal (Refereed)
    Abstract [en]

    Data show that specifically chelating lysosomal redox-active iron can prevent most H2O2-induced DNA damage. Lysosomes seem to contain the major pool of redox-active labile iron within the cell. Under oxidative stress conditions, this iron may then relocate to the nucleus and play an important role for DNA damage by taking part in Fenton reactions.

  • 42.
    Kurz, Tino
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Leake, Alan
    von Zglinicki, Thomas
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Relocalized redox-active lysosomal iron is an important mediator of oxidative-stress-induced DNA damage2004In: Biochemical Journal, ISSN 0264-6021, E-ISSN 1470-8728, Vol. 378, no 3, p. 1039-1045Article in journal (Refereed)
    Abstract [en]

    Oxidative damage to nuclear DNA is known to involve site-specific Fenton-type chemistry catalysed by redox-active iron or copper in the immediate vicinity of DNA. However, the presence of transition metals in the nucleus has not been shown convincingly. Recently, it was proposed that a major part of the cellular pool of loose iron is confined within the acidic vacuolar compartment [Yu, Persson, Eaton and Brunk (2003) Free Radical Biol. Med. 34, 1243-1252, Persson, Yu, Tirosh, Eaton and Brunk (2003) Free Radical Biol. Med. 34, 1295-1305]. Consequently, rupture of secondary lysosomes, as well as subsequent relocation of labile iron to the nucleus, could be an important intermediary step in the generation of oxidative damage to DNA. To test this concept we employed the potent iron chelator DFO (desferrioxamine) conjugated with starch to form an HMM-DFO (high-molecular-mass DFO complex). The HMM-DFO complex will enter cells only via fluid-phase endocytosis and remain within the acidic vacuolar compartment, thereby chelating redox-active iron exclusively inside the endosomal/lysosomal compartment. Both free DFO and HMM-DFO equally protected lysosomal-membrane integrity against H2O 2-induced oxidative disruption. More importantly, both forms of DFO prevented H2O2-induced strand breaks in nuclear DNA, including telomeres. To exclude the possibility that lysosomal hydrolases, rather than iron, caused the observed DNA damage, limited lysosomal rupture was induced using the lysosomotropic detergent O-methyl-serine dodecylamine hydrochloride, subsequently, hardly any DNA damage was found. These observations suggest that rapid oxidative damage to cellular DNA is minimal in the absence of redox-active iron and that oxidant-mediated DNA damage, observed in normal cells, is mainly derived from intralysosomal iron translocated to the nucleus after lysosomal rupture.

  • 43.
    Kurz, Tino
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Terman, Alexei
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Autophagy, ageing and apoptosis: The role of oxidative stress and lysosomal iron2007In: Archives of Biochemistry and Biophysics, ISSN 0003-9861, E-ISSN 1096-0384, Vol. 462, no 2, p. 220-230Article in journal (Refereed)
    Abstract [en]

    As an outcome of normal autophagic degradation of ferruginous materials, such as ferritin and mitochondrial metalloproteins, the lysosomal compartment is rich in labile iron and, therefore, sensitive to the mild oxidative stress that cells naturally experience because of their constant production of hydrogen peroxide. Diffusion of hydrogen peroxide into the lysosomes results in Fenton-type reactions with the formation of hydroxyl radicals and ensuing peroxidation of lysosomal contents with formation of lipofuscin that amasses in long-lived postmitotic cells. Lipofuscin is a non-degradable polymeric substance that forms at a rate that is inversely related to the average lifespan across species and is built up of aldehyde-linked protein residues. The normal accumulation of lipofuscin in lysosomes seems to reduce autophagic capacity of senescent postmitotic cells-probably because lipofuscin-loaded lysosomes continue to receive newly formed lysosomal enzymes, which results in lack of such enzymes for autophagy. The result is an insufficient and declining rate of autophagic turnover of worn-out and damaged cellular components that consequently accumulate in a way that upsets normal metabolism. In the event of a more substantial oxidative stress, enhanced formation of hydroxyl radicals within lysosomes jeopardizes the membrane stability of particularly iron-rich lysosomes, specifically of autophagolysosomes that have recently participated in the degradation of iron-rich materials. For some time, the rupture of a limited number of lysosomes has been recognized as an early upstream event in many cases of apoptosis, particularly oxidative stress-induced apoptosis, while necrosis results from a major lysosomal break. Consequently, the regulation of the lysosomal content of redox-active iron seems to be essential for the survival of cells both in the short- and the long-term. © 2007 Elsevier Inc. All rights reserved.

  • 44.
    Kurz, Tino
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Terman, Alexei
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Geriatric .
    Gustafsson, Bertil
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    Lysosomes and oxidative stress in aging and apoptosis2008In: Biochimica et Biophysica Acta - General Subjects, ISSN 0304-4165, E-ISSN 1872-8006, Vol. 1780, no 11, p. 1291-1303Article in journal (Refereed)
    Abstract [en]

    The lysosomal compartment consists of numerous acidic vesicles (pH ~ 4-5) that constantly fuse and divide. It receives a large number of hydrolases from the trans-Golgi network, while their substrates arrive from both the cell's outside (heterophagy) and inside (autophagy). Many macromolecules under degradation inside lysosomes contain iron that, when released in labile form, makes lysosomes sensitive to oxidative stress. The magnitude of generated lysosomal destabilization determines if reparative autophagy, apoptosis, or necrosis will follow. Apart from being an essential turnover process, autophagy is also a mechanism for cells to repair inflicted damage, and to survive temporary starvation. The inevitable diffusion of hydrogen peroxide into iron-rich lysosomes causes the slow oxidative formation of lipofuscin in long-lived postmitotic cells, where it finally occupies a substantial part of the volume of the lysosomal compartment. This seems to result in a misdirection of lysosomal enzymes away from autophagosomes, resulting in depressed autophagy and the accumulation of malfunctioning mitochondria and proteins with consequent cellular dysfunction. This scenario might put aging into the category of autophagy disorders. © 2008 Elsevier B.V. All rights reserved.

  • 45.
    Kurz, Tino
    et al.
    Linköping University, Department of Medical and Health Sciences, Division of Drug Research. Linköping University, Faculty of Health Sciences.
    Terman, Alexei
    Linköping University, Department of Clinical and Experimental Medicine, Geriatric. Linköping University, Faculty of Health Sciences.
    Gustafsson, Bertil
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Brunk, Ulf T.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Lysosomes In Iron Metabolism, Ageing And Apoptosis2008In: Histochemistry and Cell Biology, ISSN 0948-6143, E-ISSN 1432-119X, Vol. 129, no 4, p. 389-406Article in journal (Refereed)
    Abstract [en]

    The lysosomal compartment is essential for a variety of cellular functions, including the normal turnover of most long-lived proteins and all organelles. The compartment consists of numerous acidic vesicles (pH ~4-5) that constantly fuse and divide. It receives a large number of hydrolases (~50) from the trans-Golgi network, and substrates from both the cells’ outside (heterophagy) and inside (autophagy). Many macromolecules contain iron that gives rise to an iron-rich environment in lysosomes that recently have degraded such macromolecules. Iron-rich lysosomes are sensitive to oxidative stress, while ‘resting’ lysosomes, which have not recently participated in autophagic events, are not. The magnitude of oxidative stress determines the degree of lysosomal destabilization and, consequently, whether arrested growth, reparative autophagy, apoptosis, or necrosis will follow. Heterophagy is the first step in the process by which immunocompetent cells modify antigens and produce antibodies, while exocytosis of lysosomal enzymes may promote tumor invasion, angiogenesis, and metastasis. Apart from being an essential turnover process, autophagy is also a mechanism by which cells will be able to sustain temporary starvation and rid themselves of intracellular organisms that have invaded, although some pathogens have evolved mechanisms to prevent their destruction. Mutated lysosomal enzymes are the underlying cause of a number of lysosomal storage diseases involving the accumulation of materials that would be the substrate for the corresponding hydrolases, were they not defective. The normal, low-level diffusion of hydrogen peroxide into iron-rich lysosomes causes the slow formation of lipofuscin in long-lived postmitotic cells, where it occupies a substantial part of the lysosomal compartment at the end of the life span. This seems to result in the diversion of newly produced lysosomal enzymes away from autophagosomes, leading to the accumulation of malfunctioning mitochondria and proteins with consequent cellular dysfunction. If autophagy were a perfect turnover process, postmitotic ageing and several age-related neurodegenerative diseases would, perhaps, not take place.

  • 46.
    Kågedal, Katarina
    et al.
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Faculty of Health Sciences.
    Zhao, Ming
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Faculty of Health Sciences.
    Svensson, Irene
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Faculty of Health Sciences.
    Brunk, Ulf
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Faculty of Health Sciences.
    Sphingosine-induced apoptosis is dependent on lysosomal proteases2001In: Biochemical Journal, ISSN 0264-6021, E-ISSN 1470-8728, Vol. 359, no 2, p. 335-343Article in journal (Refereed)
    Abstract [en]

    We propose a new mechanism for sphingosine-induced apoptosis, involving relocation of lysosomal hydrolases to the cytosol. Owing to its lysosomotropic properties, sphingosine, which is also a detergent, especially when protonated, accumulates by proton trapping within the acidic vacuolar apparatus, where most of its action as a detergent would be exerted. When sphingosine was added in low-to-moderate concentrations to Jurkat and J774 cells, partial lysosomal rupture occurred dose-dependently, starting within a few minutes. This phenomenon preceded caspase activation, as well as changes of mitochondrial membrane potential. High sphingosine doses rapidly caused extensive lysosomal rupture and ensuing necrosis, without antecedent apoptosis or caspase activation. The sphingosine effect was prevented by pre-treatment with another, non-toxic, lysosomotropic base, ammonium chloride, at 10mM. The lysosomal protease inhibitors, pepstatin A and epoxysuccinyl-L-leucylamido-3-methyl-butane ethyl ester ('E-64d'), inhibited markedly sphingosine-induced caspase activity to almost the same degree as the general caspase inhibitor benzyloxycarbonyl-Val-Ala-DL-Asp-fluoromethylketone ('Z-VAD-FMK'), although they did not by themselves inhibit caspases. We conclude that cathepsin D and one or more cysteine proteases, such as cathepsins B or L, are important mediators of sphingosine-induced apoptosis, working upstream of the caspase cascade and mitochondrial membrane-potential changes.

  • 47.
    Li, Wei
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology.
    Hellsten, Anna
    Linköping University, Department of Neuroscience and Locomotion. Linköping University, Faculty of Health Sciences.
    Xu, Lihua
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology.
    Zhuang, D-M
    Jansson, Katarina
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Care, Pharmacology.
    Yuan, Xi Ming
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Neuroscience and Locomotion, Pathology.
    Foam cell death induced by 7β-hydroxycholesterol is mediated by labile iron-driven oxidative injury: Mechanisms underlying induction of ferritin in human atheroma2005In: Free Radical Biology & Medicine, ISSN 0891-5849, E-ISSN 1873-4596, Vol. 39, no 7, p. 864-875Article in journal (Refereed)
    Abstract [en]

    Human atherosclerotic lesions typically contain large amounts of ferritin associated with apoptotic macrophages and foam cells, although the reasons are unknown. In the present investigation, we studied the relationship between ferritin induction and occurrence of apoptosis in 7β-hydroxycholesterol (7β-OH)-treated monocytic cells and macrophages. We found that 7β-OH enlarges the intracellular labile iron pool, increases formation of reactive oxygen species (ROS), and induces ferritin and cytosolic accumulation of lipid droplets, lysosomal destabilization, and apoptototic macrophage death. Since ferritin is a phase II-type protective protein, our findings suggest that ferritin upregulation here worked as an inefficient defense mechanism. Addition to the culture medium of both a membrane-permeable iron chelator 10-phenanthroline and the non-membrane-permeable iron chelators apoferritin and desferrioxamine afforded significant protection against the 7β-OH-induced effects. Consequently, endocytosed iron compounds dramatically augmented 7β-OH-induced cytotoxicity. We conclude that oxidized lipid 7β-OH causes not only foam cell formation but also oxidative damage with abnormal metabolism of cellular iron. The findings suggest that modulation of iron metabolism in human atheroma may be a potential therapeutic strategy against atherosclerosis. © 2005 Elsevier Inc. All rights reserved.

  • 48.
    Li, Wei
    et al.
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Department of Medicine and Care, Internal Medicine. Linköping University, Faculty of Health Sciences.
    Yuan, Xi Ming
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Department of Medicine and Care, Internal Medicine. Linköping University, Faculty of Health Sciences.
    Olsson, Anders
    Linköping University, Department of Medical and Health Sciences, Internal Medicine. Linköping University, Faculty of Health Sciences.
    Brunk, Ulf
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Faculty of Health Sciences.
    Uptake of Oxidized LDL by Macrophages Results in Partial Lysosomal Enzyme Inactivation and Relocation1998In: Arteriosclerosis, Thrombosis and Vascular Biology, ISSN 1079-5642, E-ISSN 1524-4636, Vol. 18, no 2, p. 177-84Article in journal (Refereed)
    Abstract [en]

    The cytotoxicity of oxidized LDL (oxLDL) to several types of artery wall cells might contribute to atherosclerosis by causing cell death, presumably by both apoptosis and necrosis. After its uptake into macrophage lysosomes by receptor-mediated endocytosis, oxLDL is poorly degraded, resulting in ceroid-containing foam cells. We studied the influence of oxLDL on lysosomal enzyme activity and, in particular, on lysosomal membrane stability and the modulation of these cellular characteristics by HDL and vitamin E (vit-E). Unexposed cells and cells exposed to acetylated LDL (AcLDL) were used as controls. The lysosomal marker enzymes cathepsin L and N-acetyl-β-glucosaminidase (NAβGase) were biochemically assayed in J-774 cells after fractionation. Lysosomal integrity in living cells was assayed by the acridine orange (AO) relocation test. Cathepsin D was immunocytochemically demonstrated in J-774 cells and human monocyte-derived macrophages. We found that the total activities of NAβGase and cathepsin L were significantly decreased, whereas their relative cytosolic activities were enhanced, after oxLDL exposure. Labilization of the lysosomal membranes was further proven by decreased lysosomal AO uptake and relocation to the cytosol of cathepsin D, as estimated by light and electron microscopic immunocytochemistry. HDL and vit-E diminished the cytotoxicity of oxLDL by decreasing the lysosomal damage. The results indicate that endocytosed oxLDL not only partially inactivates lysosomal enzymes but also destabilizes the acidic vacuolar compartment, causing relocation of lysosomal enzymes to the cytosol. Exposure to AcLDL resulted in its uptake with enlargement of the lysosomal apparatus, but the stability of the lysosomal membranes was not changed.

  • 49.
    Li, Wei
    et al.
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Faculty of Health Sciences.
    Yuan, Ximing
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Faculty of Health Sciences.
    Brunk, Ulf
    Linköping University, Department of Neuroscience and Locomotion, Pathology. Linköping University, Faculty of Health Sciences.
    OxLDL-induced macrophage cytotoxicity is mediated by lysosomal rupture and modified by intralysosomal redox-active iron1998In: Free radical research, ISSN 1071-5762, E-ISSN 1029-2470, Vol. 29, no 5, p. 389-98Article in journal (Refereed)
    Abstract [en]

    Oxidized low density lipoprotein (oxLDL) is believed to play a central role in atherogenesis. LDL is oxidized in the arterial intima by mechanisms that are still only partially understood. OxLDL is then taken up by macrophages through scavenger receptor-mediated endocytosis, which then leads to cellular damage, including apoptosis. The complex mechanisms by which oxLDL induces cell injury are mostly unknown. This study has demonstrated that oxLDL-induced damage of macrophages is associated with iron-mediated intralysosomal oxidative reactions, which cause partial lysosomal rupture and ensuing apoptosis. This series of events can be prevented by pre-exposing cells to the iron-chelator, desferrioxamine (DFO), whereas it is augmented by pretreating the cells with a low molecular weight iron complex. Since both DFO and the iron complex would be taken up by endocytosis, and thus directed to the lysosomal compartment, the results suggest that the normal contents of lysosomal low molecular weight iron may play an important role in oxLDL-induced cell damage, presumably by catalyzing intralysosomal fragmentation of lipid peroxides and the formation of toxic aldehydes and oxygen-centered radicals.

  • 50.
    Li, Wei
    et al.
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences.
    Yuan, Ximing
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine.
    Ivanova, S.
    Laboratory of Biomedical Science, North Shore-LIE Research Institute, Manhasset, NY 11030, United States.
    Tracey, K.J.
    Laboratory of Biomedical Science, North Shore-LIE Research Institute, Manhasset, NY 11030, United States.
    Eaton, John Wallace
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Experimental Pathology . Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
    Brunk, Ulf
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    3-Aminopropanal, formed during cerebral ischaemia, is a potent lysosomotropic neurotoxin2003In: Biochemical Journal, ISSN 0264-6021, E-ISSN 1470-8728, Vol. 371, no 2, p. 429-436Article in journal (Refereed)
    Abstract [en]

    Cytotoxic polyamine-derived amino aldehydes, formed during cerebral ischaemia, damage adjacent tissue (the so-called 'penumbra') not subject to the initial ischaemic insult. One such product is 3-aminopropanal (3-AP), a potent cytotoxin that accumulates in ischaemic brain, although the precise mechanisms responsible for its formation are still unclear. More relevant to the present investigations, the mechanisms by which such a small aldehydic compound might be cytotoxic are also not known, but we hypothesized that 3-AP, having the structure of a weak lysosomotropic base, might concentrate within lysosomes, making these organelles a probable focus of initial toxicity. Indeed, 3-AP leads to lysosomal rupture of D384 glioma cells, a process which clearly precedes caspase activation and apoptotic cell death. Immunohistochemistry reveals that 3-AP concentrates in the lysosomal compartment and prevention of this accumulation by the lysosomotropic base ammonia, NH3, protects against 3-AP cytotoxicity by increasing lysosomal pH. A thiol compound, N-(2-mercaptopropionyl)glycine, reacts with and neutralizes 3-AP and significantly inhibits cytoxocity. Both amino and aldehyde functions of 3-AP are necessary for toxicity: the amino group confers lysosomotropism and the aldehyde is important for additional, presently unknown, reactions. We conclude that 3-AP exerts its toxic effects by accumulating intralysosomally, causing rupture of these organelles and releasing lysosomal enzymes which initiate caspase activation and apoptosis (or necrosis if the lysosomal rupture is extensive). These results may have implications for the development of new therapeutics designed to lessen secondary damage arising from focal cerebral ischaemia.

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