liu.seSearch for publications in DiVA
Change search
Refine search result
12345 51 - 100 of 231
CiteExportLink to result list
Permanent link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Rows per page
  • 5
  • 10
  • 20
  • 50
  • 100
  • 250
Sort
  • Standard (Relevance)
  • Author A-Ö
  • Author Ö-A
  • Title A-Ö
  • Title Ö-A
  • Publication type A-Ö
  • Publication type Ö-A
  • Issued (Oldest first)
  • Issued (Newest first)
  • Created (Oldest first)
  • Created (Newest first)
  • Last updated (Oldest first)
  • Last updated (Newest first)
  • Disputation date (earliest first)
  • Disputation date (latest first)
  • Standard (Relevance)
  • Author A-Ö
  • Author Ö-A
  • Title A-Ö
  • Title Ö-A
  • Publication type A-Ö
  • Publication type Ö-A
  • Issued (Oldest first)
  • Issued (Newest first)
  • Created (Oldest first)
  • Created (Newest first)
  • Last updated (Oldest first)
  • Last updated (Newest first)
  • Disputation date (earliest first)
  • Disputation date (latest first)
Select
The maximal number of hits you can export is 250. When you want to export more records please use the Create feeds function.
  • 51.
    Grenegård, Magnus
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    The tyrosine phosphatase inhibitor dephostatin exerts multiple effects on cytosolic calcium in human platelets1999In: Thrombosis and Haemostasis, ISSN 0340-6245, p. 1121-Conference paper (Other academic)
  • 52.
    Grenegård, Magnus
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Lindström, Eva
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Reactive oxygen species derived from platelets reduce the action of nitric oxide2007Conference paper (Other academic)
  • 53.
    Grenegård, Magnus
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Lindström, Eva
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Reactive oxygen species derived from platelets reduce the action of nitric oxide2007Conference paper (Other academic)
  • 54.
    Grenegård, Magnus
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Lindström, Eva
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Asplund-Persson, Anna
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Gunnarsson, Peter
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Mechanisms underlying platelet desensitisation towards nitric oxide2006Conference paper (Other academic)
  • 55.
    Grenegård, Magnus
    et al.
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Vretenbrant-Öberg, Karin
    Linköping University, Department of Clinical and Experimental Medicine, Clinical Chemistry. Linköping University, Faculty of Health Sciences.
    Nylander, Martina
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Désilets, Stéphanie
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Lindström, Eva G
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Larsson, Anders
    Department of Medical Sciences, University Hospital, Uppsala SE-75105, Sweden.
    Ramström, Ida
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Ramström, Sofia
    Linköping University, Department of Clinical and Experimental Medicine, Clinical Chemistry. Linköping University, Faculty of Health Sciences.
    Lindahl, Tomas L
    Linköping University, Department of Clinical and Experimental Medicine, Clinical Chemistry. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Chemistry.
    The ATP-gated P2X1 receptor plays a pivotal role in activation of aspirin-treated platelets by thrombin and epinephrine2008In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 283, no 27, p. 18493-18504Article in journal (Refereed)
    Abstract [en]

    Human platelets express protease-activated receptor 1 (PAR1) and PAR4 but limited data indicate for differences in signal transduction. We studied the involvement of PAR1 and PAR4 in the cross-talk between thrombin and epinephrine. The results show that epinephrine acted via alpha(2A)-adrenergic receptors to provoke aggregation, secretion, and Ca(2+) mobilization in aspirin-treated platelets pre-stimulated with subthreshold concentrations of thrombin. Incubating platelets with antibodies against PAR4 or the PAR4-specific inhibitor pepducin P4pal-i1 abolished the aggregation. Furthermore, platelets pre-exposed to the PAR4-activating peptide AYPGKF, but not to the PAR1-activating peptide SFLLRN, were aggregated by epinephrine, whereas both AYPGKF and SFLLRN synergized with epinephrine in the absence of aspirin. The roles of released ATP and ADP were elucidated by using antagonists of the purinergic receptors P2X(1), P2Y(1), and P2Y(12) (i.e. NF449, MRS2159, MRS2179, and cangrelor). Intriguingly, ATP, but not ADP, was required for the epinephrine/thrombin-induced aggregation. In Western blot analysis, a low concentration of AYPGKF, but not SFLLRN, stimulated phosphorylation of Akt on serine 473. Moreover, the phosphatidyl inositide 3-kinase inhibitor LY294002 antagonized the effect of epinephrine combined with thrombin or AYPGKF. Thus, in aspirin-treated platelets, PAR4, but not PAR1, interacts synergistically with alpha(2A)-adrenergic receptors, and the PI3-kinase/Akt pathway is involved in this cross-talk. Furthermore, in PAR4-pretreated platelets, epinephrine caused dense granule secretion, and subsequent signaling from the ATP-gated P2X(1)-receptor and the alpha(2A)-adrenergic receptor induced aggregation. These results suggest a new mechanism that has ATP as a key element and circumvents the action of aspirin on epinephrine-facilitated PAR4-mediated platelet activation.

  • 56.
    Gunnarsson, Peter
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Arts and Sciences.
    α1-acid glycoprotein modulates the function of human neutrophils and platelets2009Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The acute-phase protein α1-acid glycoprotein (AGP; orosomucoid) was initially identified andcharacterised in the 1950s. The normal plasma concentration is around 0.5-1 mg/ml butduring inflammation the concentration increase several fold and the carbohydrate compositionof the protein changes. AGP is a highly glycosylated protein with 45 % of the molecularweight consisting of glycans. These glycans are believed to be of importance for the functionof the protein. However, the precise physiological role of AGP is still unclear.

    The present thesis reveals that AGP at physiological concentration induce calcium elevationin human neutrophils and platelets. In neutrophils this response was enhanced several fold ifsurface L-selectin was pre-engaged. Our results showed that this L-selectin-mediatedamplification was abolished if the neutrophils were pre-treated with Src or phosphoinositide3-kinase (PI3K) inhibitors. AGP alone did not induce production of reactive oxygen species(ROS) in neutrophils. However, if the neutrophils were activated by the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP) a subsequent addition of AGP caused aprominent ROS response. Moreover, both the calcium rise and the ROS response weredepending on sialic acid residues on AGP. In the case of calcium elevation we defined thereceptor as sialic-acid-binding immunoglobulin-like lectin (Siglec)-5 on the neutrophil.

    In platelets, AGP induced a Rho-kinase dependent phosphorylation of myosin phosphatasetarget subunit-1 (MYPT1) and a minor calcium response. This resulted in a prominent plateletshape change (i.e. spherical shape and granule centralization) recorded as change in lighttransmission and by differential interference contrast (DIC) microscopy. The shape changecaused by AGP was strongly suppressed by inhibition of Rho-kinase and abolished by Rhokinaseinhibition combined with chelation of intracellular calcium. No other manifestations ofplatelet activation like aggregation or secretion were registered. Opposite to neutrophils theeffect of AGP on platelets was not mediated by an interaction between sialic acid and siglecmolecules. However, the results indicated that AGP may bind to a collagen/thrombospondin-1surface receptor. Endogenous inhibitors like nitric oxide (NO) and adenosine abolished theAGP-induced platelet shape change. The antagonizing action of NO on shape change causedby AGP was long acting. In comparison, other aspects of agonist-induced platelet activation(e.g. intracellular calcium elevations) are only transiently suppressed by NO. This indicatesthat endothelium-derived NO may play a crucial role to counter balance the effect of AGP in vivo.

    Take together the results in this thesis reveal that AGP can initiate intracellular signalling andmodulate functional responses in neutrophils and platelets.

    List of papers
    1. The acute-phase protein alpha 1-acid glycoprotein (AGP) induces rises in cytosolic Ca2+ in neutrophil granulocytes via sialic acid binding immunoglobulin-like lectins (siglecs)
    Open this publication in new window or tab >>The acute-phase protein alpha 1-acid glycoprotein (AGP) induces rises in cytosolic Ca2+ in neutrophil granulocytes via sialic acid binding immunoglobulin-like lectins (siglecs)
    2007 (English)In: The FASEB journal : official publication of the Federation of American Societies for Experimental Biology, ISSN 1530-6860, Vol. 21, no 14, p. 4059-4069Article in journal (Refereed) Published
    Abstract [en]

    We studied whether the acute-phase protein alpha1-acid glycoprotein (AGP) induces rises in [Ca2+]i in neutrophils and sought to identify the corresponding AGP receptor (or receptors). We found that AGP elicited a minimal rise in [Ca2+]i in Fura-2-loaded neutrophils, and this response was markedly enhanced by pretreatment with anti-L-selectin antibodies. (The EC50 value of the AGP-induced Ca2+ response was 9 microg/ml.) Activation of phospholipase-C, Src tyrosine kinases, and PI3 kinases proved to be essential for the AGP-mediated increase in [Ca2+]i, whereas the p38 MAPK and SYK signaling pathways were not involved. Furthermore, antibodies against sialic acid binding, immunoglobulin-like lectin 5 (Siglec-5) and oligosaccharide 3'-sialyl-lactose both antagonized the AGP-induced response and caused an immediate increase in [Ca2+]i in anti-L-selectin-treated neutrophils, which indicates a signaling capacity of Siglec-5. We used modified forms of AGP (treated with mild periodate or neuraminidase) to establish the importance of sialic acid residues. The modified forms of AGP caused a much smaller rise in [Ca2+]i than did unaltered AGP. Affinity chromatography confirmed that unchanged AGP, but not neuraminidase-treated AGP, bound to Siglec-5. Our report provides the first evidence for a signaling capacity by AGP through a defined receptor. Pre-engagement of L-selectin significantly enhanced this signaling capacity.

    Keywords
    Orosomucoid, plasma protein, calcium signaling, carbohydrate, L-selectin, phagocyte
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:liu:diva-20262 (URN)10.1096/fj.07-8534com (DOI)17675532 (PubMedID)
    Available from: 2009-09-01 Created: 2009-09-01 Last updated: 2009-09-09Bibliographically approved
    2. Sialic Acid Dependent and Independent Effects of alpha 1-Acid Glycoprotein (AGP) on Human Platelets
    Open this publication in new window or tab >>Sialic Acid Dependent and Independent Effects of alpha 1-Acid Glycoprotein (AGP) on Human Platelets
    2008 (English)In: 2008 Meeting of the Society for Glycobiology, 2008, Vol. 18, no 11, p. 990-990Conference paper, Published paper (Other academic)
    Abstract [en]

    Objective: We have recently shown that terminal sialic acid residues are essential for α1-acidglycoprotein (AGP)-induced Ca2+ mobilization in neutrophils. The aim of the present studywas to establish the importance of sialic acid-residues on AGP in modulating humanneutrophil functions, with emphasis on the generation of reactive oxygen species (ROS).Material and methods: ROS were measured by luminol-enhanced chemiluminescence inisolated human neutrophils.

    Results: We found that AGP did not provoke ROS generation in resting or L-selectin presensitizedneutrophils. Moreover, AGP did not affect the N-formyl-methionyl-leucylphenylalanine(fMLP)-induced ROS generation but it slightly suppressed opsonized zymosaninducedresponses. However, when the neutrophils were pre-stimulated with fMLP, thefollowing addition of AGP provoked a marked ROS response. Dose-response studies and timestudies revealed that the ROS generating capacity of AGP was maximal at a concentration of0.05 mg/ml and when given 3-10 min after addition of fMLP. A desialylated form of AGP orpre-treatment of neutrophils with 3’- and 6’-sialyllactose caused a substantial lower ROSresponse in neutrophils pre-stimulated with fMLP.

    Conclusions: Our data show that AGP can stimulate a second ROS response in fMLP preactivatedneutrophils and that terminal sialic acid residues on AGP play a crucial role in thisregard.

    Keywords
    α1-acid glycoprotein, neutrophils, reactive oxygen species, sialic acid
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:liu:diva-16353 (URN)
    Available from: 2009-01-16 Created: 2009-01-16 Last updated: 2009-09-09
    3. Characterisation of GEA 3175 on human platelets: comparison with S-nitroso-N-acetyl-D,L-penicillamine
    Open this publication in new window or tab >>Characterisation of GEA 3175 on human platelets: comparison with S-nitroso-N-acetyl-D,L-penicillamine
    2004 (English)In: European Journal of Pharmacology, ISSN 0014-2999, E-ISSN 1879-0712, Vol. 496, no 1-3, p. 1-9Article in journal (Refereed) Published
    Abstract [en]

    By comparing the effect of two nitric oxide (NO)-containing compounds, we found that S-nitroso-N-acetyl-d,l-penicillamine (SNAP), but not GEA 3175 (1,2,3,4-Oxatriazolium,3-(3-chloro-2-metylphenyl)-5-[[(4-methylphenyl)sulfonyl]amino]-, hydroxide inner salt), released NO. Despite this, both drugs elevated cyclic guanosine 3′,5′-monophosphate (cGMP) levels in human platelets. However, SNAP was more effective after short exposure times (5 and 20 s). The compounds also inhibited thrombin-induced rises in cytosolic Ca2+. Time studies revealed that the action of SNAP rapidly declined by increasing the length of incubation (from 5 s to 30 min). This desensibilisation phenomenon mainly involved the release of Ca2+ from intracellular stores. In comparison, GEA 3175-induced inhibition of cytosolic Ca2+ signalling was much more long-lasting. The soluble guanylyl cyclase (sGC) inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) reversed the effect of GEA 3175 on cytosolic Ca2+. Consequently, this inhibition depends solely on the increase in cGMP. In summary, differences between GEA 3175 and SNAP were observed in NO releasing, cGMP elevating and Ca2+ suppressive properties.

    Keywords
    NO (Nitric Oxide), cGMP (cyclic guanosine 3′, 5′-monophosphate), Calcium, Aggregation, Platelet, SNAP (S-nitroso-N-acetyl-d, l-penicillamine), GEA 3175
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:liu:diva-20473 (URN)10.1016/j.ejphar.2004.06.002 (DOI)15288569 (PubMedID)
    Available from: 2009-09-09 Created: 2009-09-09 Last updated: 2017-12-13Bibliographically approved
    4. α1-acid glycoprotein (AGP)-induced platelet shape change involvesthe Rho/Rho kinase signalling pathway
    Open this publication in new window or tab >>α1-acid glycoprotein (AGP)-induced platelet shape change involvesthe Rho/Rho kinase signalling pathway
    2009 (English)In: Thrombosis and Haemostasis, ISSN 0340-6245, Vol. 102, no 4, p. 694-703Article in journal (Refereed) Published
    Abstract [en]

    α1-acid glycoprotein (AGP) is an acute-phase protein that contributes to inflammation processes. The role of AGP in platelet activation and thrombosis is, however, largely unknown. Therefore, we thoroughly investigated the effects of AGP on human platelets. Platelets were isolated from healthy volunteers and subsequently exposed to AGP. Platelet responses were monitored as change in light transmission, intracellular calcium concentration, light microscopy and protein phosphorylation by Western blot. We found that AGP induced platelet shape change independently of a second release of adenine nucleotides or thromboxane A2, and that effect was abolished by endotheliumderived platelet inhibitors such as nitric oxide (NO) and adenosine. Furthermore, AGP triggered a minor calcium response and a pronounced Rho/Rho-kinase-dependent increase in Thr696 phosphorylation of myosin phosphatase target subunit 1 (MYPT1). Moreover, the Rho/Rho-kinase inhibitor Y-27632 significantly decreased the AGP-induced shape change. The results also showed that the AGP-elicited shape change was antagonised by pretreatment with low doses of collagen and thrombospondin- 1. Our results describe a novel mechanism by which AGP stimulates platelet shape change via activation of the Rho/Rhokinase signalling pathway. Physiological important platelet inhibitors, such as NO, completely counterbalance the effect of AGP. Hence, the present study indicates that AGP directly contributes to platelet activation, which in turn might have an impact in physiological haemostasis and/or pathological thrombosis.

    Place, publisher, year, edition, pages
    Stuttgart, Germany: Schattauer Gmbh, 2009
    Keywords
    shape change, Rho-kinase, Platelets, α1-acid glycoprotein
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:liu:diva-20474 (URN)10.1160/TH09-03-0156 (DOI)000271039400013 ()
    Available from: 2009-09-09 Created: 2009-09-09 Last updated: 2018-10-02Bibliographically approved
  • 57.
    Gunnarsson, Peter
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Fornander, Louise
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Påhlsson, Peter
    Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
    Grenegård, Magnus
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Sialic acid residues play a pivotal role in alpha(1)-acid glycoprotein (AGP)-induced generation of reactive oxygen species in chemotactic peptide pre-activated neutrophil granulocytes2010In: INFLAMMATION RESEARCH, ISSN 1023-3830, Vol. 59, no 2, p. 89-95Article in journal (Refereed)
    Abstract [en]

    We have recently shown that terminal sialic acid residues are essential for alpha(1)-acid glycoprotein (AGP)-induced Ca2+ mobilization in neutrophils. The aim of the present study was to establish the importance of sialic acid residues on AGP in modulating human neutrophil functions, with emphasis on the generation of reactive oxygen species (ROS). ROS were measured by luminol-enhanced chemiluminescence in isolated human neutrophils. We found that AGP did not provoke ROS generation in resting or L-selectin presensitized neutrophils. Moreover, AGP did not affect the N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced ROS generation, but it slightly suppressed opsonized zymosan-induced responses. However, when the neutrophils were prestimulated with fMLP, the subsequent addition of AGP provoked a marked ROS response. Dose-response studies and time studies revealed that the ROS generating capacity of AGP was highest at a concentration of 0.05 mg/ml and when given 3-10 min after addition of fMLP. A desialylated form of AGP or pretreatment of neutrophils with 3- and 6-sialyllactose caused a substantially lower ROS response in neutrophils prestimulated with fMLP. Our data show that AGP can stimulate a second ROS response in fMLP preactivated neutrophils and that terminal sialic acid residues on AGP play a crucial role in this regard.

  • 58.
    Gunnarsson, Peter
    et al.
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Levander, Louise
    Linköping University, Department of Biomedicine and Surgery, Cell biology. Linköping University, Faculty of Health Sciences.
    Påhlsson, Peter
    Linköping University, Department of Biomedicine and Surgery, Cell biology. Linköping University, Faculty of Health Sciences.
    Grenegård, Magnus
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    The acute-phase protein alpha 1-acid glycoprotein (AGP) induces rises in cytosolic Ca2+ in neutrophil granulocytes via sialic acid binding immunoglobulin-like lectins (siglecs)2007In: The FASEB journal : official publication of the Federation of American Societies for Experimental Biology, ISSN 1530-6860, Vol. 21, no 14, p. 4059-4069Article in journal (Refereed)
    Abstract [en]

    We studied whether the acute-phase protein alpha1-acid glycoprotein (AGP) induces rises in [Ca2+]i in neutrophils and sought to identify the corresponding AGP receptor (or receptors). We found that AGP elicited a minimal rise in [Ca2+]i in Fura-2-loaded neutrophils, and this response was markedly enhanced by pretreatment with anti-L-selectin antibodies. (The EC50 value of the AGP-induced Ca2+ response was 9 microg/ml.) Activation of phospholipase-C, Src tyrosine kinases, and PI3 kinases proved to be essential for the AGP-mediated increase in [Ca2+]i, whereas the p38 MAPK and SYK signaling pathways were not involved. Furthermore, antibodies against sialic acid binding, immunoglobulin-like lectin 5 (Siglec-5) and oligosaccharide 3'-sialyl-lactose both antagonized the AGP-induced response and caused an immediate increase in [Ca2+]i in anti-L-selectin-treated neutrophils, which indicates a signaling capacity of Siglec-5. We used modified forms of AGP (treated with mild periodate or neuraminidase) to establish the importance of sialic acid residues. The modified forms of AGP caused a much smaller rise in [Ca2+]i than did unaltered AGP. Affinity chromatography confirmed that unchanged AGP, but not neuraminidase-treated AGP, bound to Siglec-5. Our report provides the first evidence for a signaling capacity by AGP through a defined receptor. Pre-engagement of L-selectin significantly enhanced this signaling capacity.

  • 59.
    Gunnarsson, Peter
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Levander, Louise
    Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
    Påhlsson, Peter
    Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
    Grenegård, Magnus
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    α1-acid glycoprotein (AGP)-induced platelet shape change involvesthe Rho/Rho kinase signalling pathway2009In: Thrombosis and Haemostasis, ISSN 0340-6245, Vol. 102, no 4, p. 694-703Article in journal (Refereed)
    Abstract [en]

    α1-acid glycoprotein (AGP) is an acute-phase protein that contributes to inflammation processes. The role of AGP in platelet activation and thrombosis is, however, largely unknown. Therefore, we thoroughly investigated the effects of AGP on human platelets. Platelets were isolated from healthy volunteers and subsequently exposed to AGP. Platelet responses were monitored as change in light transmission, intracellular calcium concentration, light microscopy and protein phosphorylation by Western blot. We found that AGP induced platelet shape change independently of a second release of adenine nucleotides or thromboxane A2, and that effect was abolished by endotheliumderived platelet inhibitors such as nitric oxide (NO) and adenosine. Furthermore, AGP triggered a minor calcium response and a pronounced Rho/Rho-kinase-dependent increase in Thr696 phosphorylation of myosin phosphatase target subunit 1 (MYPT1). Moreover, the Rho/Rho-kinase inhibitor Y-27632 significantly decreased the AGP-induced shape change. The results also showed that the AGP-elicited shape change was antagonised by pretreatment with low doses of collagen and thrombospondin- 1. Our results describe a novel mechanism by which AGP stimulates platelet shape change via activation of the Rho/Rhokinase signalling pathway. Physiological important platelet inhibitors, such as NO, completely counterbalance the effect of AGP. Hence, the present study indicates that AGP directly contributes to platelet activation, which in turn might have an impact in physiological haemostasis and/or pathological thrombosis.

  • 60.
    Gyllensten, Hanna
    et al.
    Nordic School Public Health NHV, Sweden .
    Hakkarainen, Katja M.
    Nordic School Public Health NHV, Sweden .
    Jonsson, Anna K.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Andersson Sundell, Karolina
    Nordic School Public Health NHV, Sweden .
    Hägg, Staffan
    Linköping University, Department of Medical and Health Sciences, Clinical Pharmacology. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre for Diagnostics, Department of Clinical Pharmacology.
    Rehnberg, Clas
    Karolinska Institute, Sweden .
    Carlsten, Anders
    Nordic School Public Health NHV, Sweden Medical Prod Agency, Sweden .
    Modelling drug-related morbidity in Sweden using an expert panel of pharmacists2012In: INTERNATIONAL JOURNAL OF CLINICAL PHARMACY, ISSN 2210-7703, Vol. 34, no 4, p. 538-546Article in journal (Refereed)
    Abstract [en]

    Background Drug-related morbidity (DRM) is common and to some extent preventable, and associated with considerable costs in patients attending hospital. In outpatients and in the general public corresponding data are limited, but pharmacists expert opinion has suggested high rates of DRM also in US ambulatory care. It is unknown if the results are applicable in Sweden today. Objective To estimate the proportions of patients with DRM and preventable DRM and the cost-of-illness (COI) of DRM in Sweden based on pharmacists expert opinion. Setting Swedish healthcare. Method The study applied a conceptual model of DRM based on a decision tree. An expert panel of pharmacists determined the probabilities of therapeutic outcomes of medication therapy. The COI analysis included direct costs from the healthcare perspective. Sensitivity analyses were performed for variations in probabilities and pathway costs. Main outcome measure DRM included new medical problems (adverse drug reactions, drug dependence and intoxications) and therapeutic failures (insufficient effects of medicines and morbidity due to untreated indication). Results The expert panel estimated that 61 +/- A 14 % (mean +/- A SD) of all patients attending healthcare suffered from DRM, of which 29 +/- A 8 % suffered from new medical problems, 18 +/- A 6 % from therapeutic failures, and 15 +/- A 7 % from a combination of both. The DRM was considered preventable in 45 +/- A 15 % of the patients with DRM. The estimated COI was EUR 997 per patient attending healthcare, corresponding to an annual cost of EUR 6,600 million to the Swedish healthcare system. The COI ranged from EUR 490 to EUR 1,314 when varying the participants probabilities of DRM and clinical outcomes from the first to the third quartile. Of the pathway costs, the COI was most sensitive to variation in the cost of prolonged hospital stay (COI range EUR 953-1,306). Conclusion According to pharmacists expert opinion, a large proportion of patients in Sweden experience DRM and preventable DRM, and the estimated COI of DRM is extensive. Since observational studies have not addressed the burden of DRM to the general public, this study adds the pharmacists perception on DRM. Other healthcare professionals perceptions on DRM need to be investigated in future studies.

  • 61.
    Hallbäck, Ida
    et al.
    Östergötlands Läns Landsting, Center for Diagnostics, Department of Clinical Pharmacology. Linköping University, Department of Medical and Health Sciences. Linköping University, Faculty of Health Sciences.
    Hägg, Staffan
    Linköping University, Department of Medical and Health Sciences, Clinical Pharmacology. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Center for Diagnostics, Department of Clinical Pharmacology.
    Eriksson, Andreas
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Whiss, Per
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    In vitro effects of serotonin and noradrenaline reuptake inhibitors on human platelet adhesion and coagulation2012In: Pharmacological Reports, ISSN 1734-1140, Vol. 64, no 4, p. 979-983Article in journal (Refereed)
    Abstract [en]

    Background: Although several studies show that there is an increased risk of bleeding events during antidepressant treatment with selective serotonin reuptake inhibitors (SSRIs), few studies show direct effects in vitro of SSRIs on hemostasis. less thanbrgreater than less thanbrgreater thanMethods: This study was undertaken to investigate the effects on platelet adhesion and plasma coagulation (APTT and PT) of two common SSRIs, citalopram and sertraline, the selective noradrenaline reuptake inhibitor reboxetine, and the serotonin and noradrenaline reuptake inhibitor venlafaxine. less thanbrgreater than less thanbrgreater thanResults: None of the compounds affected plasma coagulation significantly but all compounds except for venlafaxine inhibited platelet adhesion by approximately 50% or more at the highest concentration (100 mu g/l, p andlt; 0.01). The potency of respective compound to inhibit platelet adhesion to both collagen and fibrinogen surfaces was in the following order; citalopram andgt; sertraline andgt; reboxetine. In contrast, venlafaxine caused a weak but statistically significant increased platelet adhesion to fibrinogen. less thanbrgreater than less thanbrgreater thanConclusion: This study showed that sertraline, citalopram and reboxetine direct and acutely decrease platelet adhesion to both collagen and fibrinogen in vitro. These results also indicate that increased risk for bleeding complications in antidepressant users may not only be explained by depletion of serotonin in platelets.

  • 62.
    Hammarström, Per
    et al.
    Linköping University, Department of Physics, Chemistry and Biology, Biochemistry. Linköping University, The Institute of Technology.
    Ali Malik, Muhammad
    Linköping University, Department of Physics, Chemistry and Biology. Linköping University, The Institute of Technology.
    Mishra, Rajesh
    Linköping University, Department of Physics, Chemistry and Biology, Biochemistry. Linköping University, The Institute of Technology.
    Svensson, Samuel
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Tengvall, Pentti
    Linköping University, Department of Physics, Chemistry and Biology, Applied Physics . Linköping University, The Institute of Technology.
    Lundström, Ingemar
    Linköping University, Department of Physics, Chemistry and Biology, Applied Physics . Linköping University, The Institute of Technology.
    A catalytic surface for amyloid fibril formation2008In: Journal of Physics, Conference Series, ISSN 1742-6588, E-ISSN 1742-6596, Vol. 100Article in journal (Refereed)
    Abstract [en]

    A hydrophobic surface incubated in a solution of protein molecules (insulin monomers) was made into a catalytic surface for amyloid fibril formation by repeatedly incubate, rinse and dry the surface. The present contribution describes how this unexpected transformation occurred and its relation to rapid fibrillation of insulin solutions in contact with the surface. A tentative model of the properties of the catalytic surface is given, corroborated by ellipsometric measurements of the thickness of the organic layer on the surface and by atomic force microscopy. The surfaces used were spontaneously oxidized silicon made hydrophobic through treatment in dichlorodimethylsilane.

  • 63.
    Hedlund, Petter
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Editorial Comment on Characteristics of Spontaneous Activity in the Bladder Trigone: in EUROPEAN UROLOGY, vol 56, issue 2, pg 354.2009In: European Urology, ISSN 0302-2838, E-ISSN 1873-7560, Vol. 56, no 2, p. 354-354Article in journal (Other academic)
    Abstract [en]

    n/a

  • 64.
    Henricson, Joakim
    et al.
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences.
    Droog Tesselaar, Erik
    Linköping University, Department of Biomedical Engineering. Linköping University, The Institute of Technology.
    Persson, Karin
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Nilsson, Gert
    Linköping University, Department of Biomedical Engineering, Biomedical Instrumentation. Linköping University, The Institute of Technology.
    Sjöberg, Folke
    Linköping University, Department of Clinical and Experimental Medicine, Burn Unit . Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of Plastic Surgery, Hand surgery UHL.
    Assessment of microvascular function by study of the dose‐response effects of iontophoretically applied drugs (acetylcholine and sodium nitroprusside): Methods and comparison with in vitro studies2007In: Microvascular Research, ISSN 0026-2862, E-ISSN 1095-9319, Vol. 73, no 2, p. 143-149Article in journal (Refereed)
    Abstract [en]

    Current knowledge about vascular function stems mainly from pharmacological in vitro studies using mounted vascular strips on a strain gauge. We know of no paper that has systematically examined the possibility of assessing the conventional dose–response effects of iontophoresis and laser Doppler investigation of vasoactive substances and compared those relations to data obtained from strips mounted on a strain gauge.

    We used the vasoactive substances acetylcholine (endothelium dependent) and sodium nitroprusside (endothelium independent) and an antagonist (atropine) to enable further investigations in the receptor physiology of iontophoresis.

    Dose–response curves from the iontophoresis experiments showed close similarity to those obtained by vascular strips mounted on a strain gauge. The coefficient of variation (CV) of the dose–response factors found in iontophoresis (both inter and intra experimental variability) was low. The iontophoretic effective dose of 50% (ED50) for acetylcholine and nitroprusside had only CVs of 25% and 26%, respectively, compared with 71% and 77% for the vascular strips. Acetylcholine-induced response was antagonized by iontophoresis of atropine. Contrary to expectations, this antagonism was not competitive.

    The results show that iontophoresis in combination with laser Doppler technology produces reproducible and reliable dose–response curves that picture the vascular effects of vasoactive drugs.

  • 65.
    Jacobsson, Leif S.
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Persson, Karin
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Aberg, Gunnar
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Andersson, Rolf G. G.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Karlberg, Bengt E.
    Linköping University, Department of Medicine and Care, Internal Medicine. Linköping University, Faculty of Health Sciences.
    Olsson, Anders G.
    Linköping University, Department of Medicine and Care, Internal Medicine. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Local Health Care Services in Central Östergötland, Department of Endocrinology and Gastroenterology.
    Antiatherosclerotic Effects of the Angiotensin-Converting Enzyme Inhibitors Captopril and Fosinopril in Hypercholesterolemic Minipigs1994In: Journal of Cardiovascular Pharmacology, ISSN 0160-2446, E-ISSN 1533-4023, Vol. 24, no 4, p. 670-677Article in journal (Refereed)
    Abstract [en]

    We evaluated the two angiotensin-converting enzyme (ACE) inhibitors captopril and fosinopril with regard to possible antiatherosclerotic effects in minipigs. Experimental hypercholesterolemia and atherosclerosis was produced in 33 minipigs of the Gottingen strain by an egg yolk/cholesterol-enriched diet for 1 year. One group (n = 11) was fed the atherogenic diet alone and served as a control. A second group (n = 11) received captopril (80 mg/kg/day) added to the atherogenic diet, and a third group (n = 11) was treated in the same manner but with fosinopril (8 mg/kglday). The drug treatments produced significant reduction in serum ACE activity associated with a reactive increase in plasma renin activity (PRA), but had only minor effects on plasma lipids and lipoproteins. At the end of the treatment period, all animals were killed and examined for degree of atherosclerosis. The percentage of atherosclerotic area in the abdominal aorta was significantly lower in both drug-treated groups as compared with controls. Furthermore, accumulation of cholesterol in the thoracic and abdominal aorta was inhibited by drug treatment. Finally, the percentage of intimal thickening in abdominal aorta was significantly reduced in the drug-treated groups. In conclusion, the ACE inhibitors captopril and fosinopril inhibited development of atherosclerosis in hypercholesterolemic minipigs.

  • 66. Johansson, H
    et al.
    Bengtsson, Torbjörn
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    Platelets and plasma modulate oxygen radical production in neutrophils by mechanisms involving adenosine and arachidonic acid metabolites2001In: Molecular Biology of the Cell, ISSN 1059-1524, E-ISSN 1939-4586, Vol. 12, p. 1415-Conference paper (Other academic)
  • 67.
    Josefsson, Martin
    et al.
    Linköping University, Department of Physics, Chemistry and Biology. Linköping University, The Institute of Technology.
    Rydberg, Irene
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Determination of methylphenidate and ritalinic acid in blood, plasma and oral fluid from adolescents and adults using protein precipitation and liquid chromatography tandem mass spectrometry-A method applied on clinical and forensic investigations2011In: JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, ISSN 0731-7085, Vol. 55, no 5, p. 1050-1059Article in journal (Refereed)
    Abstract [en]

    A validated, accurate and sensitive LC-MS/MS method for determination of racemic methylphenidate and its metabolite ritalinic acid has been developed. The analytes were quantified by tandem mass spectrometry operating in positive electrospray ionization mode with multiple reaction monitoring. Blood, plasma and oral fluid samples of 100 mu l were prepared by simple precipitation with 200 RI of an aqueous solution of zinc sulphate in methanol. Corresponding deuterated internal standards were used for quantification. Calibrations for methylphenidate and ritalinic acid were linear within the selected range of 0.2-30 ng/ml and 10-1500 ng/ml in blood or plasma and in the range of 1-500 ng/ml and 0.25-125 ng/ml in oral fluid, respectively. The method was successfully applied for the analysis of samples from patients treated with methylphenidate in the dose range of 36-72 mg/day and some representative ante mortem and post mortem samples from clinical and forensic toxicological investigations. A three to fourfold higher concentration of methylphenidate was found in oral fluid compared with blood while for ritalinic acid the concentrations were about 25-fold lower in oral fluid.

  • 68.
    Junker, J P E
    et al.
    Harvard University.
    Lonnqvist, Susanna
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences.
    Karlsson, L K
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences.
    Grenegard, Magnus
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Kratz, Gunnar
    Linköping University, Department of Clinical and Experimental Medicine, Plastic Surgery, Hand Surgery and Burns. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Sinnescentrum, Department of Plastic Surgery, Hand surgery UHL.
    Endothelial differentiation of human dermal fibroblasts in JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, vol 6, issue SI, pp 149-1492012In: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, John Wiley and Sons , 2012, Vol. 6, no SI, p. 149-149Conference paper (Refereed)
    Abstract [en]

    n/a

  • 69.
    Junker, J P
    et al.
    Harvard University.
    Lönnqvist, Susanna
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences.
    Karlsson, L K
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences.
    Grenegård, Magnus
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Kratz, Gunnar
    Linköping University, Department of Clinical and Experimental Medicine, Plastic Surgery, Hand Surgery and Burns. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Sinnescentrum, Department of Plastic Surgery, Hand surgery UHL.
    ENDOTHELIAL DIFFERENTIATION OF HUMAN DERMAL FIBROBLASTS in WOUND REPAIR AND REGENERATION, vol 20, issue 2, pp A27-A272012In: WOUND REPAIR AND REGENERATION, Wiley-Blackwell , 2012, Vol. 20, no 2, p. A27-A27Conference paper (Refereed)
    Abstract [en]

    n/a

  • 70.
    Kambiz Fotoohi, Alan
    et al.
    Karolinska Institute.
    Coulthard, Sally A
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Albertioni, Freidoun
    Karolinska Institute.
    Thiopurines: Factors influencing toxicity and response2010In: BIOCHEMICAL PHARMACOLOGY, ISSN 0006-2952, Vol. 79, no 9, p. 1211-1220Article in journal (Refereed)
    Abstract [en]

    Thiopurines are the backbone of current anti-leukemia regimens and have also been effective immunosuppressive agents for the past half a century. Extensive research on their mechanism of action has been undertaken, yet many issues remain to be addressed to resolve unexplained cases of thiopurine toxicity or treatment failure. The aim of this review is to summarize current knowledge of the mechanism of thiopurine action in experimental models and put into context with clinical observations. Clear understanding of their metabolism will contribute to maximizing efficacy and minimizing toxicity by individually tailoring therapy according to the expression profile of relevant factors involved in thiopurine activation pathway.

  • 71.
    Karlsson, Annika M.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Signaling for color change in melanophores: and a biosensor application2001Doctoral thesis, monograph (Other academic)
    Abstract [en]

    Melanophores are dark brown pigment cells located in the skin of fish, amphibia, reptiles, and many invertebrates. The color of the animal can change via rearrangement of pigment granules, melanosomes, in the cells. The dark melanophores can either hide colorful cells so that the animal appears dark, or let through colors from underneath. The animal regulates its colors and patterns via communicating nerve cells and hormones in the blood stream. It is nowadays well established that melatonin-stimulation of melanophores results in aggregation of melanosomes to the cell center and that the evident outcome is more transparent cells. It has previously been shown that the activity of serine and threonine kinases as well as phosphatases regulates the distribution of melanosomes in the cells.

    We wanted to study if tyrosine phosphorylations were involved in the regulation of melanosome aggregation. Melatonin-stimulated signaling in the African clawed frog, Xenopus laevis, melanophores was examined. Melansome aggregation was accompanied by tyrosine phosphorylation as shown by immunoblots. Inhibition of tyrosine phosphorylation reduced melanosome aggregation by melatonin, and the phosphorylation most likely regulated pigment aggregation. Tyrosine phosphorylation of the protein was mediated via a Gi/o protein coupled receptor, probably the melatonin receptor Mel1c. The phosphorylation was most likely not a result of the classical Gi/o protein pathway, as Src-kinase and mitogen-activated protein kinase seemed required for phosphorylation and melanosome aggregation. Two candidates for the phosphorylated protein were presented, talin and β-spectrin.

    The possible involvement of nitric oxide in melanosome aggregation by melatonin was investigated. Nitric oxide appeared to be necessary for melanosome aggregation. The effect of nitric oxide synthase inhibition on melanosome aggregation was not mediated via changes in the tyrosine-phosphorylated protein. We speculated that nitric oxide could affect melanosome distribution via modifications of the actin cytoskeleton. The use of recombinant melanophores as a biosensor has also been examined. A human G protein coupled receptor, opioid receptor 3, was inserted into melanophores by electroporation. The transfected melanophores responded dose-dependently to opioids and an inhibitor of opioid receptors reduced the aggregation response. Future melanophore biosensors migh detect a variety of substances, such as narcotics, pheromones, odors, and tastes.

  • 72.
    Karlsson, Annika M.
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Bjuhr, Katarina
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Testorf, Martin
    Linköping University, Department of Biomedical Engineering, Physiological Measurements. Linköping University, The Institute of Technology.
    Öberg, Åke
    Linköping University, Department of Biomedical Engineering, Physiological Measurements. Linköping University, The Institute of Technology.
    Lerner, Ethan
    Bunsen Rush Laboratories, Dallas, TX, USA.
    Lundström, Ingemar
    Linköping University, Department of Physics, Chemistry and Biology, Applied Physics. Linköping University, The Institute of Technology.
    Svensson, Samuel P.S.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Biosensing of opioids using frog melanophores2002In: Biosensors and Bioelectronics, ISSN 0956-5663, Vol. 17, no 4, p. 331-335Article in journal (Refereed)
    Abstract [en]

    Spectacular color changes of fishes, frogs and other lower vertebrates are due to the motile activities of specialized pigment containing cells. Pigment cells are interesting for biosensing purposes since they provide an easily monitored physiological phenomenon. Melanophores, containing dark brown melanin pigment granules, constitute an important class of chromatophores. Their melanin-filled pigment granules may be stimulated to undergo rapid dispersion throughout the melanophores (cells appear dark), or aggregation to the center of the melanophores (cells appear light). This simple physiological response can easily be measured in a photometer. Selected G protein coupled receptors can be functionally expressed in cultured frog melanophores. Here, we demonstrate the use of recombinant frog melanophores as a biosensor for the detection of opioids. Melanophores were transfected with the human opioid receptor 3 and used for opiate detection. The response to the opioid receptor agonist morphine and a synthetic opioid peptide was analyzed by absorbance readings in an aggregation assay. It was shown that both agonists caused aggregation of pigment granules in the melanophores, and the cells appeared lighter. The pharmacology of the expressed receptors was very similar to its mammalian counterpart, as evidenced by competitive inhibition by increasing concentrations of the opioid receptor inhibitor naloxone. Transfection of melanophores with selected receptors enables the creation of numerous melanophore biosensors, which respond selectively to certain substances. The melanophore biosensor has potential use for measurement of substances in body fluids such as saliva, blood plasma and urine.

  • 73.
    Karlsson, Annika M.
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Lerner, Michael R.
    Department of Dermatology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
    Unett, David
    Department of Dermatology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
    Lundström, Ingemar
    Linköping University, Department of Physics, Chemistry and Biology, Applied Physics. Linköping University, The Institute of Technology.
    Svensson, Samuel P.S.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Melatonin-induced organelle movement in melanophores is coupled to tyrosine phosphorylation of a high molecularweight protein2000In: Cellular signalling, ISSN 0898-6568, Vol. 12, no 7, p. 469-474Article in journal (Refereed)
    Abstract [en]

    Melanophores, brown to black pigment cells from, for example, Xenopus laevis, contain mobile melanin filled organelles, and are well suited for studies on organelle movement. The intracellular regulation of the movement seems to be controlled by serine and threonine phosphorylations and dephosphorylations. Melatonin induces aggregation of the melanosomes to the cell centre through a Gi/o-protein-coupled receptor, Mel1c, which leads to an inhibition of PKA and a stimulation of PP2A. However, this study shows that the melatonin-induced aggregation of melanosomes is also accompanied by tyrosine phosphorylation of a protein with a molecular weight of 280 kDa. Cells pre-incubated with genistein, an inhibitor of tyrosine phosphorylations, showed inhibited melanosome movement after melatonin stimulation, and a lower degree of tyrosine phosphorylation of the 280 kDa protein. The adenylyl cyclase activator forskolin, and the Gi/o protein inhibitor pertussis toxin, also inhibited tyrosine phosphorylation of the 280 kDa protein. The results indicate that melatonin stimulation generates tyrosine phosphorylation of a high molecular weight protein, an event that seems to be essential for melanosome aggregation.

  • 74.
    Karlsson, Jan O.G.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Antioxidant activity of mangafodipir is not a new finding [4] (multiple letters)2004Other (Other academic)
    Abstract [en]

    [No abstract available]

  • 75.
    Karlsson, Jan Olof
    et al.
    Linköping University, Department of Medical and Health Sciences. Linköping University, Faculty of Health Sciences.
    Adolfsson, Karin
    County Hospital Ryhov, Sweden .
    Thelin, Bo
    County Hospital Ryhov, Sweden .
    Jynge, Per
    Linköping University, Department of Medical and Health Sciences, Division of Drug Research. Linköping University, Faculty of Health Sciences.
    Andersson, Rolf
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Falkmer, Ursula G
    Linköping University, Department of Medical and Health Sciences. Linköping University, Faculty of Health Sciences.
    First Clinical Experience with the Magnetic Resonance Imaging Contrast Agent and Superoxide Dismutase Mimetic Mangafodipir as an Adjunct in Cancer Chemotherapy-A Translational Study2012In: Translational Oncology, ISSN 1944-7124, E-ISSN 1936-5233, Vol. 5, no 1, p. 32-38Article in journal (Refereed)
    Abstract [en]

    Preclinical research suggests that the clinically approved magnetic resonance imaging contrast agent mangafodipir may protect against adverse events (AEs) caused by chemotherapy, without interfering negatively with the anticancer efficacy. The present translational study tested if pretreatment with mangafodipir lowers AEs during curative (adjuvant) FOLFOX6 chemotherapy in stage III colon cancer (Dukes C). The study was originally scheduled to include 20 patients, but because of the unforeseen withdrawal of mangafodipir from the market, the study had to be closed after 14 patients had been included. The withdrawal of mangafodipir was purely based on commercial considerations from the producer and not on any safety concerns. The patients were treated throughout the first 3 of 12 scheduled cycles. Patients were randomized to a 5-minute infusion of either mangafodipir or placebo (7 in each group). AEs were evaluated according to the National Cancer Institutes (NCI) Common Terminology Criteria for Adverse Events and the Sanofi-NCI criteria. The primary end points were neutropenia and neurosensory toxicity. There were four AEs of grade 3 (severe) and one AE of grade 4 (life threatening) in four patients in the placebo group, whereas there were none in the mangafodipir group (P andlt; .05). Of the grade 3 and 4 events, two were neutropenia and one was neurosensory toxicity. Furthermore, white blood cell count was statistically, significantly higher in the mangafodipir group than in the placebo group (P andlt; .01) after treatment with FOLFOX. This small feasibility study seems to confirm what has been demonstrated preclinically, namely, that pretreatment with mangafodipir lowers AEs during adjuvant 5-fluorouracil plus oxaliplatin-based chemotherapy in colon cancer patients.

  • 76.
    Karlsson, Jan Olof G
    et al.
    PledPharma AB, Sweden .
    Kurz, Tino
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Flechsig, Susanne
    Expt Pharmacol and Oncology Berlin Buch GmbH, Germany .
    Nasstrom, Jacques
    PledPharma AB, Sweden .
    Andersson, Rolf
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Superior Therapeutic Index of Calmangafodipir in Comparison to Mangafodipir as a Chemotherapy Adjunct2012In: TRANSLATIONAL ONCOLOGY, ISSN 1944-7124, Vol. 5, no 6, p. 492-502Article in journal (Refereed)
    Abstract [en]

    Mangafodipir is a magnetic resonance imaging contrast agent with manganese superoxide dismutase (MnSOD) mimetic activity. The MnSOD mimetic activity protects healthy cells against oxidative stress-induced detrimental effects, e. g., myelosuppressive effects of chemotherapy drugs. The contrast property depends on in vivo dissociation of Mn2+ from mangafodipir-about 80% dissociates after injection. The SOD mimetic activity, however, depends on the intact Mn complex. Complexed Mn2+ is readily excreted in the urine, whereas dissociated Mn2+ is excreted slowly via the biliary route. Mn is an essential but also a potentially neurotoxic metal. For more frequent therapeutic use, neurotoxicity due to Mn accumulation in the brain may represent a serious problem. Replacement of 4/5 of Mn2+ in mangafodipir with Ca2+ (resulting in calmangafodipir) stabilizes it from releasing Mn2+ after administration, which roughly doubles renal excretion of Mn. A considerable part of Mn2+ release from mangafodipir is governed by the presence of a limited amount of plasma zinc (Zn2+). Zn2+ has roughly 10(3) and 10(9) times higher affinity than Mn2+ and Ca2+, respectively, for fodipir. Replacement of 80% of Mn2+ with Ca2+ is enough for binding a considerable amount of the readily available plasma Zn2+, resulting in considerably less Mn2+ release and retention in the brain and other organs. At equivalent Mn2+ doses, calmangafodipir was significantly more efficacious than mangafodipir to protect BALB/c mice against myelosuppressive effects of the chemotherapy drug oxaliplatin. Calmangafodipir did not interfere negatively with the antitumor activity of oxaliplatin in CT26 tumor-bearing syngenic BALB/c mice, contrary calmangafodipir increased the antitumor activity. Translational Oncology (2012) 5, 492-502

  • 77.
    Karlsson, Lisa K.
    et al.
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences.
    Junker, Johan
    Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
    Grenegård, Magnus
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Kratz, Gunnar
    Linköping University, Department of Clinical and Experimental Medicine, Burn Unit . Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of Plastic Surgery, Hand surgery UHL.
    Human Dermal Fibroblasts and Single-Cell Clone Fibroblasts Have theCapacity to Alter Their Phenotype Towardsan Endothelial-Like Cell type2009In: European Cells and Materials, ISSN 1473-2262, E-ISSN 1473-2262Article in journal (Other academic)
    Abstract [en]

    We investigated the capacity of normal human dermal fibroblasts to alter their phenotype into an endothelialcell-like phenotype. By utilising in vitro cell culture models, the part played by different types of serum andmedium constituents in inducing a phenotypic change of fibroblasts was investigated. The experiments usedprimary cultures of human endothelial cells, human dermal fibroblasts and single-cell clone fibroblasts. Thelatter cell type was obtained by clonal expansion using a micromanipulator technique. The results showed thatthe presence of human serum in the cell culture medium caused both types of fibroblasts to express vonWillebrand factor, to incorporate fluorochrome-labelled LDL, and to start forming capillary-like networks in asimilar way to endothelial cells. The phenotypic shift was detectable after 4 days of cell culture and reached amaximum after 7-10 days. To our knowledge this is the first report to describe differentiation of humanfibroblasts towards an endothelial cell-like phenotype. The results also show that the underlying mechanism ofthe phenotypic shift is a change in gene expression in the dermal fibroblasts and not fusion between different celltypes. Collectively, the present results indicate that human dermal fibroblasts may be a novel cell source forcreating vascular endothelium.

  • 78.
    Karlsson, Lisa K
    et al.
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences.
    Junker, Johan P E
    Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
    Grenegård, Magnus
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Kratz, Gunnar
    Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of Plastic Surgery, Hand surgery UHL.
    Human Dermal Fibroblasts: A Potential Cell Source for Endothelialization of Vascular Grafts2009In: Annals of Vascular Surgery, ISSN 0890-5096, E-ISSN 1615-5947, Vol. 23, no 5, p. 663-674Article in journal (Refereed)
    Abstract [en]

    BACKGROUND: Recently, there has been an intense ongoing search for suitable cell sources for vascular tissue engineering. Previous studies report that cells with multilineage potential have been found within the connective stroma of the skin. In line with this, preliminary data from our group suggest that human dermal fibroblasts have the capacity to alter their phenotype into an endothelial cell-like phenotype in vitro. As a first step in using these cells in vascular tissue engineering, we investigated their ability to form an endothelial cell-like layer on a scaffold in vitro. Furthermore, we studied the possibility of seeding dermal fibroblasts on a scaffold and later commencing with induction toward an endothelial cell-like phenotype. METHODS: Cells cultured in either normal fibroblast medium or endothelial induction medium were seeded on a gelatin-based scaffold. To study the organization of cells, routine staining was performed. Differentiation was confirmed by Western blotting and immunohistochemistry with antibodies directed toward molecules commonly used to identify endothelial cells. RESULTS AND CONCLUSION: Our data support that human dermal fibroblasts differentiated toward endothelial cell-like cells prior to seeding showed histological resemblance to mature endothelial cells, while fibroblasts seeded and later induced into endothelial differentiation grew in multilayer. However, expression of various surface molecules indicative of an endothelial phenotype was seen using both techniques. In conclusion, the results presented in this study indicate that human dermal fibroblasts differentiated toward an endothelial cell-like phenotype may be a novel cell source for endothelialization of vascular grafts.

  • 79.
    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.

  • 80.
    Kingbäck, Maria
    et al.
    Linköping University, Department of Medical and Health Sciences, Clinical Pharmacology. Linköping University, Faculty of Health Sciences.
    Josefsson, M
    Linköping University, Department of Medical and Health Sciences, Pharmacology.
    Kugelberg, Fredrik
    Linköping University, Department of Medical and Health Sciences, Clinical Pharmacology. Linköping University, Faculty of Health Sciences.
    Bengtsson, Finn
    Linköping University, Department of Medical and Health Sciences, Clinical Pharmacology. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre for Diagnostics, Department of Clinical Pharmacology.
    Ahlner, Johan
    Linköping University, Department of Medical and Health Sciences, Forensic Science and Toxicology . Linköping University, Faculty of Health Sciences.
    Carlsson, Björn
    Linköping University, Department of Medical and Health Sciences, Clinical Pharmacology. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre for Diagnostics, Department of Clinical Pharmacology.
    Stereoselective analysis of venlafaxine and its three major metabolites by liquid chromatography with electrospray tandem mass spectrometry2008Conference paper (Other academic)
  • 81. Kjellgren, K.I.
    et al.
    Svensson, Samuel
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Pharmacology .
    Ahlner, Johan
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Clinical Pharmacology .
    Saljo, R.
    Antihypertensive treatment and patient autonomy - The follow-up appointment as a resource for care2000In: Patient Education and Counseling, ISSN 0738-3991, E-ISSN 1873-5134, Vol. 40, no 1, p. 39-49Article in journal (Refereed)
    Abstract [en]

    Since hypertension is a chronic condition which generally requires long-term commitment to pharmacological therapy as well as alterations of patient lifestyle, the patient-physician communication in the clinical setting is an important determinant of the quality of care and health outcome. The aim of the present study was to explore the structure and content of the communication between the patient and the physician, and the process of decision-making at a routine follow-up appointment for hypertension. The study was based on 51 audio-recordings of authentic consultations. Most patients had a passive role in the consultations, and initiated few topics of conversation. The few topics that the patients initiated were usually not about hypertension. Patients' questions about medication mainly referred to unwanted effects of the drugs. Little time was invested in discussing risks related to hypertension. A collaborative shared decision-making was seldom observed in the consultations. Copyright (C) 2000 Elsevier Science Ireland Ltd.

  • 82.
    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.

  • 83.
    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 .
    S. B. Clark, Robert
    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 .
    Demarchi, Francesca
    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 .
    E. Palmer, Glen
    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 .
    Zhu, Xiongwei
    Case Western Reserve University, OH 44106 USA .
    L. Deter, Russell
    Baylor University, TX 77030 USA .
    Tabas, Ira
    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.

  • 84.
    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.

  • 85.
    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.

  • 86.
    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.

  • 87.
    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.

  • 88.
    Kurz, Tino
    et al.
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Grant, Derek
    GE Healthcare, Oslo, Norway.
    Andersson, Rolf
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Robertson, Towart
    Audacter Consulting, Helensburgh, Scotland.
    De Cesare, Michelandrea
    Fondazione IRCCS Istituto Nazionale Tumori di Milano, Milano, Italy.
    Karlsson, Jan Olof G.
    Linköping University, Department of Medical and Health Sciences, Clinical Pharmacology. Linköping University, Faculty of Health Sciences.
    Effects of MnDPDP andICRF-187 on Doxorubicin-Induced Cardiotoxicityand Anticancer Activity12012In: Translational Oncology, ISSN 1944-7124, E-ISSN 1936-5233, Vol. 5, no 4, p. 252-259Article in journal (Refereed)
    Abstract [en]

    Oxidative stress participates in doxorubicin (Dx)–induced cardiotoxicity. The metal complex MnDPDP and its metaboliteMnPLED possess SOD-mimetic activity, DPDP and PLED have, in addition, high affinity for iron. Mice wereinjected intravenously with MnDPDP, DPDP, or dexrazoxane (ICRF-187). Thirty minutes later, mice were killed, theleft atria were hung in organ baths and electrically stimulated, saline or Dx was added, and the contractility wasmeasured for 60 minutes. In parallel experiments, 10 μM MnDPDP or MnPLED was added directly into the organbath. The effect of MnDPDP on antitumor activity of Dx against two human tumor xenografts (MX-1 and A2780)was investigated. The in vitro cytotoxic activity was studied by co-incubating A2780 cells with MnDPDP, DPDP,and/or Dx. Dx caused a marked reduction in contractile force. In vivo treatment with MnDPDP and ICRF-187 attenuatedthe negative effect of Dx. When added directly into the bath, MnDPDP did not protect, whereas MnPLEDattenuated the Dx effect by approximately 50%. MnDPDP or ICRF-187 did not interfere negatively with the antitumoractivity of Dx, either in vivo or in vitro. Micromolar concentrations of DPDP but not MnDPDP displayed anin vitro cytotoxic activity against A2780 cells. The present results show that MnDPDP, after being metabolized toMnPLED, protects against acute Dx cardiotoxicity. Both in vivo and in vitro experiments show that cardioprotectiontakes place without interfering negatively with the anticancer activity of Dx. Furthermore, the results suggest thatthe previously described cytotoxic in vivo activity of MnDPDP is an inherent property of DPDP.

    Translational Oncology (2012) 5, 252–259

  • 89.
    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.

  • 90.
    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.

  • 91.
    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)
  • 92.
    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.

  • 93.
    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.

  • 94.
    Kälvegren, Hanna
    et al.
    Linköping University, Department of Medical and Health Sciences, Division of Cardiovascular Medicine. Linköping University, Faculty of Health Sciences.
    Andersson, Johanna
    Grenegård, Magnus
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Bengtsson, Torbjörn
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Platelet activation triggered by Chlamydia pneumoniae is antagonized by 12-lipoxygenase inhibitors but not cyclooxygenase inhibitors.2007In: European Journal of Pharmacology, ISSN 0014-2999, E-ISSN 1879-0712, Vol. 566, no 1-3, p. 20-27Article in journal (Refereed)
    Abstract [en]

    Chlamydia pneumoniae is a respiratory pathogen that has been linked to cardiovascular disease. We have recently shown that C. pneumoniae activates platelets, leading to oxidation of low-density lipoproteins. The aim of the present study was to evaluate the inhibitory effects of different pharmacological agents on platelet aggregation and secretion induced by C. pneumoniae.

    Platelet interaction with C. pneumoniae was studied by analyzing platelet aggregation and ATP-secretion with Lumi-aggregometry.

    Platelet aggregation and ATP-secretion induced by C. pneumoniae was markedly inhibited by the NO-donor S-nitroso-N-acetyl-d,l-penicillamine (SNAP), an effect that was counteracted by the guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ). Pre-treatment of platelets with the 12-lipoxygenase (12-LOX) inhibitors cinnamyl-3,4-dihydroxy-α-cyanocinnamate (CDC) and 5,6,7-trikydroxyflavone (baicalein) completely blocked the activation, whereas the cyclooxygenase (COX) inhibitors 2-acetyloxybenzoic acid (aspirin) and (8E)-8-[hydroxy-(pyridin-2-ylamino)methylidene]-9-methyl-10,10-dioxo-10$l^(6)thia-9-azabicyclo[4.4.0]deca-1,3,5-trien-7-one (piroxicam) had no inhibitory effects. Opposite to C. pneumoniae-induced activation, platelets stimulated by collagen were inhibited by the COX-inhibitors but were unaffected by the 12-LOX-inhibitors. The platelet activating factor (PAF) antagonist Ginkgolide B blocked the C. pneumoniae-induced platelet activation, whereas the responses to collagen were unaffected. Furthermore, the P2Y1 and P2Y12 purinergic receptor antagonists 2'-deoxy-N6-methyladenosine 3',5'-bisphosphate (MRS2179) and N(6)-(2-methyl-thioethyl)-2-(3,3,3-trifluoropropylthio)-beta,gamma-dichloromethylene-ATP (cangrelor) inhibited the aggregation and secretion caused by C. pneumoniae.

    It is well-known that the efficacy of COX inhibitors in the prevention and treatment of cardiovascular disease varies between different patients, and that patients with low responses to aspirin have a higher risk to encounter cardiovascular events. The findings in this study showing that platelets stimulated by C. pneumoniae are unaffected by COX inhibitors but sensitive to 12-LOX inhibitors, may thus be of importance in future management of atherosclerosis and thrombosis.

  • 95.
    Kälvegren, Hanna
    et al.
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Bylin, Helena
    Linköping University, Department of Clinical and Experimental Medicine, Occupational and Environmental Medicine . Linköping University, Faculty of Health Sciences.
    Leanderson, Per
    Linköping University, Department of Clinical and Experimental Medicine, Occupational and Environmental Medicine . Linköping University, Faculty of Health Sciences.
    Richter, Arina
    Linköping University, Department of Medicine and Health Sciences, Cardiology . Linköping University, Faculty of Health Sciences.
    Grenegård, Magnus
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Bengtsson, Torbjörn
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Chlamydia pneumoniae induces nitric oxide synthase and lipoxygenase-dependent production of reactive oxygen species in platelets — effects on oxidation of low-density lipoproteins.2005In: Thrombosis and Haemostasis, ISSN 0340-6245, Vol. 94, no 2, p. 327-335Article in journal (Refereed)
    Abstract [en]

    There is increasing evidence that Chlamydia pneumoniae is linked to atherosclerosis and thrombosis. In this regard, we have recently shown that C. pneumoniae stimulates platelet aggregation and secretion, which may play an important role in the progress of atherosclerosis and in thrombotic vascular occlusion. The aims of the present study were to investigate the effects of C. pneumoniae on platelet-mediated formation of reactive oxygen species (ROS) and oxidation of low-density lipoprotein (LDL) in vitro. ROS production was registered as changes in 2´,7`-dichlorofluorescin- fluorescence in platelets with flow cytometry. LDL-oxidation was determined by measuring thiobarbituric acid reactive substances (TBARs). We found that C. pneumoniae stimulated platelet production of ROS.Polymyxin B treatment of C. pneumoniae, but not elevated temperature, abolished the stimulatory effects on platelet ROS- production, which suggests that chlamydial lipopolysaccharide has an important role. In hibition of nitric oxide synthase with nitro-L-arginine, lipoxygenase with 5,8,11-eicosatriynoic acid and protein kinase C with GF 109203X significantly lowered the production of radicals. In contrast, inhibition of NADPH-oxidase with di-phenyleneiodonium (DPI) did not affect the C. pneumoniae induced ROS-production. These findings suggest that the activities of nitric oxide synthase and lipoxygenase are the sources for ROS and that the generation is dependent of the activity of protein kinase C.The C. pneumoniae-induced ROS-production in platelets was associated with an extensive oxidation of LDL, which was significantly higher compared to the effect obtained by separate exposure of LDL to C. pneumoniae or platelets. In conclusion, C. pneumoniae interaction with platelets leading to aggregation, ROS-production and oxidative damage on LDL, may play a crucial role in the development of atherosclerotic cardiovascular disease.

  • 96.
    Kälvegren, Hanna
    et al.
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Fridfeldt (Berggren), Jonna
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Garvin, Peter
    Linköping University, Department of Medicine and Health Sciences, Division of Preventive and Social Medicine and Public Health Science. Linköping University, Faculty of Health Sciences.
    Wind, Lena
    Leanderson, Per
    Linköping University, Department of Clinical and Experimental Medicine, Occupational and Environmental Medicine . Linköping University, Faculty of Health Sciences.
    Kristenson, Margaretha
    Linköping University, Department of Medicine and Health Sciences, Division of Preventive and Social Medicine and Public Health Science. Linköping University, Faculty of Health Sciences.
    Kihlström, Erik
    Linköping University, Department of Clinical and Experimental Medicine, Clinical Microbiology . Linköping University, Faculty of Health Sciences.
    Bengtsson, Torbjörn
    Linköping University, Department of Medicine and Health Sciences, Pharmacology . Linköping University, Faculty of Health Sciences.
    Richter, Arina
    Linköping University, Department of Medicine and Health Sciences, Cardiology . Linköping University, Faculty of Health Sciences.
    Correlation between rises in Chlamydia pneumoniae-specific antibodies, platelet activation and lipid peroxidation after percutaneous coronary intervention.2008In: European Journal of Clinical Microbiology and Infectious Diseases, ISSN 0934-9723, E-ISSN 1435-4373, Vol. 27, no 7, p. 503-511Article in journal (Refereed)
    Abstract [en]

    We recently showed that Chlamydia pneumoniae activates platelets in vitro, with an associated oxidation of low-density lipoproteins. The aim of this study was to investigate whether C. pneumoniae is released during percutaneous coronary intervention (PCI) and, thereby, causes platelet activation and lipid peroxidation. Seventy-three patients undergoing coronary angiography and following PCI or coronary artery bypass graft (CABG) and 57 controls were included in the study. C. pneumoniae antibodies, serotonin and lipid peroxidation were measured before and 24 h, 1 month and 6 months after angiography. The results show that serum C. pneumoniae IgA concentrations were significantly higher in patients than in the controls. Furthermore, in 38% of the C. pneumoniae IgG positive patients, the C. pneumoniae IgG concentration increased 1 month after PCI. The levels of C. pneumoniae IgG antibodies 1 month after PCI correlated with plasma-lipid peroxidation (r = 0.91, P < 0.0001) and platelet-derived serotonin (r = 0.62, P = 0.02). There was no elevation in the total serum IgG 1 month after PCI. In conclusion, the present results suggest that PCI treatment of coronary stenosis releases C. pneumoniae from the atherosclerotic lesions, which leads to platelet activation and lipid peroxidation.

  • 97.
    Kälvegren, Hanna
    et al.
    Linköping University, Department of Medical and Health Sciences, Division of Cardiovascular Medicine. Linköping University, Faculty of Health Sciences.
    Fridfeldt, Jonna
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Bengtsson, Torbjörn
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    The role of plasma adenosine deaminase in chemoattractant-stimulated oxygen radical production in neutrophils2010In: EUROPEAN JOURNAL OF CELL BIOLOGY, ISSN 0171-9335, Vol. 89, no 6, p. 462-467Article in journal (Refereed)
    Abstract [en]

    Objectives: Adenosine deaminase (ADA) has a role in many immunity mediated disorders, such as asthma, tuberculosis and coronary artery disease. This study aims to investigate the ability of plasma ADA to modulate reactive oxygen species (ROS) production in neutrophils, and examine the involvement of adenosine and the cyclic AMP signaling pathway in this process. Methods: Neutrophils were stimulated, in the absence or presence of plasma, with the chemotactic peptide fMLP (formyl-methionyl-leucyl-phenylalanine), and the ROS production was determined with luminol-enhanced chemiluminescence. Activity of ADA was measured spectrophotometrically. Results: Plasma dose-dependently amplified the ROS generation in fMLP-stimulated neutrophils. In parallel, incubation of neutrophils in plasma elevated the total ADA-activity approximately 10 times from 1.3 U/ml to 12 U/ml. Inhibition of ADA, or type IV phosphodiesterases, significantly lowered the plasma-mediated ROS production. Furthermore, the high-affinity adenosine A(1) receptor antagonists DPCPX and 8-phenyltheophylline markedly inhibited the plasma-induced respiratory burst in neutrophils, suggesting an AI receptor-mediated mechanism. Conclusions: This study suggests that plasma ADA amplifies the release of toxic oxygen radicals from neutrophils through a downregulation of the inhibitory adenosine/cAMP-system and an enhanced activation of the stimulatory adenosine A(1)-receptor. This mechanism has probably a crucial role in regulating neutrophil function and in the defence against microbial infections. However, a sustained neutrophil activation could also contribute to inflammatory disorders such as atherosclerosis.

  • 98.
    Kälvegren, Hanna
    et al.
    Linköping University, Department of Medical and Health Sciences. Linköping University, Faculty of Health Sciences.
    Majeed, Meytham
    Linköping University, Department of health and environment.
    Bengtsson, Torbjörn
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Chlamydia pneumoniae binds to platelets and triggers P-selectin expression and aggregation: A causal role in cardiovascular disease?2003In: Arteriosclerosis, Thrombosis and Vascular Biology, ISSN 1079-5642, E-ISSN 1524-4636, Vol. 23, no 9, p. 1677-1683Article in journal (Refereed)
    Abstract [en]

    Objective - Evidence linking Chlamydia pneumoniae to atherosclerotic cardiovascular disease is expanding. Platelets are considered to play an essential role in cardiovascular diseases, however, so far platelets have not been associated with an infectious cause of atherosclerosis. This study aims to clarify the interaction between Cpneumoniae and platelets and possibly present a novel mechanism in the pathogenesis of atherosclerosis.

    Methods and Results - The effects of C pneumoniae on platelet aggregation and secretion were assessed with lumiaggregometry, and the ability of C pneumoniae to bind to platelets and stimulate expression of P-selectin was analyzed with flow cytometry. We found that Cpneumoniae, at a chlamydia:platelet ratio of 1:15, adheres to platelets and triggers P-selectin expression after 1 minute and causes an extensive aggregation and ATP secretion after 20 minutes of incubation. Inhibition of glycoprotein IIb/IIIa with Arg-Gly-Asp-Ser or abciximab markedly reduced C pneumoniae-induced platelet aggregation. Exposure of C pneumoniae to polymyxin B, but not elevated temperature, abolished the stimulatory effects on platelet activation, suggesting that chlamydial lipopolysaccharide has an active role. In contrast, other tested bacteria had no or only moderate effects on platelet functions.

    Conclusion - Our findings demonstrate a new concept of how C pneumoniae activates platelets and thereby may cause atherosclerosis and thrombotic vascular occlusion.

     

  • 99.
    Kälvegren, Hanna
    et al.
    Linköping University, Department of Medical and Health Sciences, Division of Cardiovascular Medicine. Linköping University, Faculty of Health Sciences.
    Skoglund, Caroline
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Helldahl, Christian
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Lerm, Maria
    Linköping University, Department of Clinical and Experimental Medicine, Medical Microbiology. Linköping University, Faculty of Health Sciences.
    Grenegård, Magnus
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Bengtsson, Torbjörn
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Toll-like receptor 2 stimulation of platelets is mediated by purinergic P2X1-dependent Ca2+ mobilisation, cyclooxygenase and purinergic P2Y1 and P2Y12 receptor activation2010In: Thrombosis and Haemostasis, ISSN 0340-6245, Vol. 103, no 2, p. 398-407Article in journal (Refereed)
    Abstract [en]

    Toll-like receptor 2 (TLR2), which recognise and respond to conserved microbial pathogen-associated molecular patterns, is expressed on the platelet surface. Furthermore, it has recently been shown that the TLR2/1 agonist Pam(3)CSK(4) stimulates platelet activation. The aim of the present study was to clarify important signalling events in Pam(3)CSK(4)-induced platelet aggregation and secretion. Platelet interaction with Pam(3)CSK(4) and the TLR2/6 agonist MALP-2 was studied by analysing aggregation, ATP-secretion, [Ca2+](i) mobilisation and thromboxane B2 (TxB(2)) production. The results show that Pam(3)CSK(4) but not MALP-2 induces [Ca2+](i) increase, TxB(2) production, dense granule secretion and platelet aggregation. Preincubation of platelets with MALP-2 inhibited the Pam(3)CSK(4)-induced responses. The ATP-secretion and aggregation in Pam(3)CSK(4)-stimulated platelets was impeded by the purinergic P2X(1) inhibitor MRS 2159, the purinergic P2Y(1) and P2Y(12) antagonists MRS 2179 and cangrelor, the phospholipase C inhibitor U73122, the calcium chelator BAPT-AM and aspirin. The calcium mobilisation was lowered by MRS 2159, aspirin and U73122 whereas the TxB(2) production was antagonised by MRS 2159, aspirin and BAPT-AM. When investigating the involvement of the myeloid differentiation factor-88 (MyD88) -dependent pathway, we found that platelets express MyD88 and interleukin 1 receptor-associated kinase (IRAK-1), which are proteins important in TLR signalling. However, Pam(3)CSK(4) did not stimulate a rapid (within 10 minutes) phosphorylation of IRAK-1 in platelets. In conclusion, the results show that Pam(3)CSK(4)-induced platelet aggregation and secretion depends on a P2X(1)-mediated Ca2+ mobilisation, production of TxA(2) and ADP receptor activation. The findings in this study further support a role for platelets in sensing bacterial components.

  • 100.
    Laskar, Amit
    et al.
    Linköping University, Department of Clinical and Experimental Medicine, Experimental Pathology. Linköping University, Faculty of Health Sciences.
    Andersson, Rolf G G
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Li, Wei
    Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
    Fodipir (Dp-dp) and its dephosphorylated derivative PLED are involved in mangafodipir mediated cyto-protection against 7β-hydroxycholesterol induced cell death2013Manuscript (preprint) (Other academic)
    Abstract [en]

    Mangafodipir exerts pharmacological effects, including vascular relaxation and protection against oxidative stress and cell death induced by oxysterols. Additionally, mangafodipir has been proposed for cardiovascular imaging. The primary metabolite of mangafodipir, manganese dipyridoxyl ethyldiamine (MnPLED) and its constituent, dipyridoxyl diphosphate (Dp-dp) also known as fodipir, are pharmacologically active. However, whether they affect oxysterol induced cytotoxicity is currently unknown. In this study, we examine whether the mangafodipir metabolite affects 7β-hydroxycholesterol (7βOH) induced cell death and identify the underlying mechanisms. U937 cells were pre-treated or not with mangafodipir substrate (Ms) (200 μm), MnPLED (100 μM) or Dp-dp (100 μM) for 8 hours and then exposed to 7βOH (28 μM) for 18 hours. Our results revealed that pre-treatment with MnPLED or Dp-dp protected against 7βOH induced cellular reactive oxygen species (ROS) production, apoptosis, and lysosomal membrane permeabilization (LMP). MnPLED and Dpdp in par with Ms, confer protection against 7βOH induced cytotoxicity by reducing  cellular ROS and stabilization of lysosomal membrane. These results suggest that, fodipir is the active part in mangafodipir, which shows the noted effects and its activity is conserved in MnPLED. These results further confirm the cyto-protective effect of mangafodipir and justify its potential use as a cyto-protective adjuvant.

12345 51 - 100 of 231
CiteExportLink to result list
Permanent link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf