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Cited by 633Neutrophil, Myeloperoxidase and Oxidative MechanismsRedox biology and oxidative stressImmune Response and Inflammation

Functions of ROS in Macrophages and Antimicrobial Immunity

Open access
Herb, Marc, Schramm, Michael·2021·Antioxidants
DOI 10.3390/antiox10020313PMC PMC7923022
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Abstract

Reactive oxygen species (ROS) are a chemically defined group of reactive molecules derived from molecular oxygen. ROS are involved in a plethora of processes in cells in all domains of life, ranging from bacteria, plants and animals, including humans. The importance of ROS for macrophage-mediated immunity is unquestioned. Their functions comprise direct antimicrobial activity against bacteria and parasites as well as redox-regulation of immune signaling and induction of inflammasome activation. However, only a few studies have performed in-depth ROS analyses and even fewer have identified the precise redox-regulated target molecules. In this review, we will give a brief introduction to ROS and their sources in macrophages, summarize the versatile roles of ROS in direct and indirect antimicrobial immune defense, and provide an overview of commonly used ROS probes, scavengers and inhibitors.

AI snapshot

Written from the paper’s full text.

TL;DR

This review synthesizes how macrophages use reactive oxygen species (ROS) from NADPH oxidases and mitochondria for direct antimicrobial killing and for redox regulation of immune signaling and inflammasome activation. It stresses that ROS functions are compartment- and subspecies-specific and that many studies lack mechanistic redox targets or use nonspecific tools.

Key findings

  • Nox2-derived extracellular/phagosomal ROS are central for antibacterial defense: Nox2-deficient macrophages fail to produce extracellular ROS after bacterial infection, and chronic granulomatous disease patients with Nox2/subunit mutations are more susceptible to infections.
  • Mitochondria are a major ROS source: complex III produces cytosolic mtROS that can regulate proinflammatory signaling via IKKγ/NEMO disulfide linkage, while matrix mtROS contribute to killing of some pathogens and to NLRP3/AIM2 inflammasome activation; matrix mtROS cannot escape intact mitochondria.
  • ROS also modulate inflammasomes: H2O2 at 10 mM activates NLRP3 in THP-1 cells via TXNIP–thioredoxin dissociation, and human CGD cells lacking Nox2 subunits show no defect in caspase-1/IL-1β/IL-18 after typical stimuli, whereas mouse Nox4 and mtROS have been implicated.
  • The review catalogs ROS probes, scavengers, and inhibitors and warns that total-cellular probes, global scavengers, and nonspecific inhibitors such as DPI and apocynin can mislead; compartment-specific tools and genetic evidence are needed.

Methods

  • Narrative review; no primary experiments, systematic search strategy, or inclusion criteria are stated.
  • Synthesizes literature on ROS chemistry and sources, macrophage antimicrobial functions, immune regulation, inflammasomes, and ROS detection/scavenging/inhibition tools.
  • Main analysis is qualitative synthesis and recommendations; no statistical analysis is reported.

Limitations

  • Stated: only a few studies have performed in-depth ROS analyses, and even fewer have identified precise redox-regulated target molecules.
  • Stated: common experimental pitfalls include using only one ROS probe without rationale, unspecific ROS-source inhibitors or only global scavengers, no genetic evidence for the ROS source, and inconsistent stimuli/macrophage cell types within one study.
  • Evident from the text: as a narrative review, it does not report a systematic literature search or quantitative synthesis, and several cited findings are described as controversial, correlative, or lacking mechanistic proof.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.3390/antiox10020313

Cites

417 of 417 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 369 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Cytosolic ROS levels remained unaffected in macrophages and microglia after vitamin C treatment.

    “Cytosolic ROS levels remained unaffected in macrophages and microglia after vitamin C treatment.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: Vitamin C and its reduced form DHA are cell-impermeable and imported into the cell via Na+ cotransporters in the plasma membrane.
  • Findingown data

    The authors could exclusively observe extracellular ROS scavenging in macrophages (own unpublished data) and microglia.

    “we could exclusively observe extracellular ROS scavenging in macrophages (own unpublished data) and microglia [99].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [99] Wolf, 2020, The tspo-nox1 axis controls phagocyte-triggered pathological angiogenesis in the eye, Nat. Commun. source

      Not checked in the free run.

    • Rests on an assumption: Vitamin C and its reduced form DHA are cell-impermeable and imported into the cell via Na+ cotransporters in the plasma membrane.
  • Premisecitation

    Excessive H2O2 production leads to irreversible oxidation of the sulfenic group to sulfinic and sulfonic groups, resulting in protein malfunction.

    “Excessive H2O2 production, however, leads to further oxidation of the sulfenic group (Cys-SOH) to sulfinic (Cys-SO2H) and sulfonic groups (Cys-SO3H), a process that is irreversible and results in protein malfunction [55].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [55] Winterbourn, 2008, Thiol chemistry and specificity in redox signaling, Free Radic. Biol. Med. source

      Not checked in the free run.

  • Premisecitation

    A tightly controlled increase in cellular ROS levels can have beneficial consequences for the cell, a condition termed oxidative eustress.

    “Moreover, growing evidence suggests that a tightly controlled increase in cellular ROS levels can actually have beneficial consequences for the cell, a condition that has been termed oxidative eustress [29,75].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [29] Sies, 2020, Reactive oxygen species (ros) as pleiotropic physiological signalling agents, Nat. Rev. Mol. Cell Biol. source

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    • cites [75] Niki, 2016, Oxidative stress and antioxidants: Distress or eustress?, Arch. Biochem. Biophys. source

      Not checked in the free run.

  • Premisecitation

    The Nox enzyme family and the respiratory electron transport chain of mitochondria are the two major sources of ROS in many eukaryotic cells.

    “The enzyme family of NADPH oxidases (Nox) [13,83] and the respiratory electron transport chain (ETC) of mitochondria [84,85,86] are the two major sources of ROS in many eukaryotic cells.”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [13] Lambeth, 2014, Nox enzymes and new thinking on reactive oxygen: A double-edged sword revisited, Annu. Rev. Pathol. source

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    • cites [83] Bedard, 2007, The nox family of ros-generating nadph oxidases: Physiology and pathophysiology, Physiol. Rev. source

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    • cites [84] West, 2011, Mitochondria in innate immune responses, Nat. Rev. Immunol. source

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    • cites [85] Brand, 2010, The sites and topology of mitochondrial superoxide production, Exp. Gerontol. source

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    • cites [86] Murphy, 2009, How mitochondria produce reactive oxygen species, Biochem. J. source

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  • Classified as probably not a substantial claim. 25
  • May combine two claims. 101
  • The quote only partly states this claim. 1

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    ROS levels that exceed the capacity of the cellular antioxidant defense system induce oxidative stress.

    “ROS levels that exceed the capacity of the cellular antioxidant defense system induce oxidative stress [29,75].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [29] Sies, 2020, Reactive oxygen species (ros) as pleiotropic physiological signalling agents, Nat. Rev. Mol. Cell Biol. source

      Not checked in the free run.

    • cites [75] Niki, 2016, Oxidative stress and antioxidants: Distress or eustress?, Arch. Biochem. Biophys. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationnone given

    ROS functions in macrophage-mediated immunity comprise direct antimicrobial activity against bacteria and parasites, redox-regulation of immune signaling, and induction of inflammasome activation.

    “Their functions comprise direct antimicrobial activity against bacteria and parasites as well as redox-regulation of immune signaling and induction of inflammasome activation.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Excessive oxidative stress causes damage to cellular components such as proteins, DNA and lipids and results in a number of different pathologies.

    “Excessive oxidative stress causes damage to cellular components such as proteins, DNA and lipids and results in a number of different pathologies [76,77,78].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [76] Thimmulappa, 2006, Nrf2 is a critical regulator of the innate immune response and survival during experimental sepsis, J. Clin. Investig. source

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    • cites [77] Rangasamy, 2005, Disruption of nrf2 enhances susceptibility to severe airway inflammation and asthma in mice, J. Exp. Med. source

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    • cites [78] Cross, 1987, Oxygen radicals and human disease, Ann. Intern. Med. source

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    Premisecitation

    Only a few studies investigated the role of xanthine oxidase in macrophages and suggested involvement in inflammasome activation, regulation of cytokine expression and defense against parasitic infection.

    “however, only a few studies investigated its role in macrophages and suggested involvement in inflammasome activation [144], regulation of cytokine expression [145] and defense against parasitic infection [146].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [144] Ives, 2015, Xanthine oxidoreductase regulates macrophage il1beta secretion upon nlrp3 inflammasome activation, Nat. Commun. source

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    • cites [145] Kim, 2014, The xanthine oxidase-nfat5 pathway regulates macrophage activation and tlr-induced inflammatory arthritis, Eur. J. Immunol. source

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    • cites [146] Ty, 2019, Malaria inflammation by xanthine oxidase-produced reactive oxygen species, EMBO Mol. Med. source

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  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Complex II-fueled mtROS production at complex I is still controversial and could be tissue-specific.

    “This complex II-fueled mtROS production at complex I is still controversial and could be tissue-specific [108].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [108] Neuzil, 2013, Reactive oxygen species are generated by the respiratory complex ii--evidence for lack of contribution of the reverse electron flow in complex i, FEBS J. source

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    Premisecitation

    Nox2 can produce large quantities of ROS, which are a crucial component of the antimicrobial activity of professional phagocytes, including macrophages.

    “In particular Nox2 can produce large quantities of ROS, which are a crucial component of the antimicrobial activity of professional phagocytes, including macrophages [58,93].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [58] Gluschko, 2018, The beta2 integrin mac-1 induces protective lc3-associated phagocytosis of listeria monocytogenes, Cell Host Microbe source

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    • cites [93] Pollock, 1995, Mouse model of x-linked chronic granulomatous disease, an inherited defect in phagocyte superoxide production, Nat. Genet. source

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    Premisereasoning

    Nox2-derived ROS and matrix mtROS were suggested to induce COX, but this suggestion was based exclusively on data obtained with DPI and is not fully conclusive.

    “Nox2-derived ROS [185] and matrix mtROS [186] were suggested to induce COX. However, this suggestion was based exclusively on data obtained with diphenyl iodinium (DPI), a general flavoprotein inhibitor with various side effects and with lack of specificity (see Section 7). Therefore, the findings are not fully conclusive.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [185] Barbieri, 2003, Reactive oxygen species mediate cyclooxygenase-2 induction during monocyte to macrophage differentiation: Critical role of nadph oxidase, Cardiovasc. Res. source

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    • cites [186] Kiritoshi, 2003, Reactive oxygen species from mitochondria induce cyclooxygenase-2 gene expression in human mesangial cells: Potential role in diabetic nephropathy, Diabetes source

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  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Xanthine oxidase generates H2O2 as a byproduct during oxidation of hypoxanthine and xanthine to uric acid.

    “It catalyzes the oxidation of the bases hypoxanthine and xanthine to uric acid, which is the end product of purine catabolism. H2O2 is generated as a byproduct during two oxidation steps of the enzymatic reaction, marking XO as a potential ROS source.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [137] Duley, 2020, Xanthine oxidase-lactoperoxidase system and innate immunity: Biochemical actions and physiological roles, Redox Biol. source

      Not checked in the free run.

    • cites [138] Harris, 1997, The reaction of reduced xanthine dehydrogenase with molecular oxygen. Reaction kinetics and measurement of superoxide radical, J. Biol. Chem. source

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    • cites [139] Huber, 1996, A structure-based catalytic mechanism for the xanthine oxidase family of molybdenum enzymes, Proc. Natl. Acad. Sci. USA source

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    Premisecitation

    Peroxisomes function as a ROS sink for the cytosol and regulate cellular redox balance and redox-dependent signaling.

    “With high concentrations of catalase, glutathione oxidase and SOD, peroxisomes do not only detoxify ROS produced by themselves, but also function as a ROS sink for the cytosol [150,151,152] suggesting a role for peroxisomes in regulation of the redox balance of the cell and therefore in redox-dependent signaling [148,153].”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [150] Schumann, 2008, Special delivery from mitochondria to peroxisomes, Trends Cell Biol. source

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    • cites [151] Lismont, 2015, Redox interplay between mitochondria and peroxisomes, Front. Cell Dev. Biol. source

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    • cites [152] Fransen, 2012, Role of peroxisomes in ros/rns-metabolism: Implications for human disease, Biochim. Biophys. Acta source

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    • cites [148] Fransen, M., Lismont, C., and Walton, P. (2017). The peroxisome-mitochondria connection: How and why?. Int. J. Mol. Sci., 18. source

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    • cites [153] Pan, 2020, Peroxisomes: Versatile organelles with diverse roles in plants, New Phytol. source

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    Premisecitation

    Reports of roles for CYP enzymes in inflammatory responses of macrophages are rare and address regulatory intermediates generated by CYP rather than the role of CYP-mediated ROS generation.

    “however, reports of roles for CYP enzymes in inflammatory responses of macrophages are rare [217,218] and address the regulatory intermediates generated by CYP rather than the role of CYP-mediated ROS generation.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [217] Chen, 2017, Cyp4a in tumor-associated macrophages promotes pre-metastatic niche formation and metastasis, Oncogene source

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    • cites [218] Tian, 2020, Cytochrome p450 1a1 enhances inflammatory responses and impedes phagocytosis of bacteria in macrophages during sepsis, Cell Commun. Signal. source

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    Premisecitation

    Peroxisomes generate O2− and subsequently H2O2 as byproducts during β-oxidation of fatty acids.

    “During the β-oxidation of fatty acids, O2●− and, subsequently, H2O2 are generated as byproducts [147].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [147] Shai, 2018, Systematic mapping of contact sites reveals tethers and a function for the peroxisome-mitochondria contact, Nat. Commun. source

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    Premisecitation

    Only one study investigated involvement of peroxisome-dependent ROS production in macrophage-mediated immunity and suggested a role in direct antibacterial defense and phagocytosis in Drosophila melanogaster.

    “However, so far, only one study investigated involvement of peroxisome-dependent ROS production in macrophage-mediated immunity and suggested a role in direct antibacterial defense and phagocytosis in the fruit fly Drosophila melanogaster [165].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [165] Sheshachalam, 2017, Peroxisome-mediated metabolism is required for immune response to microbial infection, Immunity source

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    Premisecitation

    Expression of various CYP enzymes in macrophages was shown over 20 years ago.

    “Expression of various CYP enzymes in macrophages was shown over 20 years ago [216]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [216] Baron, 1998, Cytochrome p450 1b1: A major p450 isoenzyme in human blood monocytes and macrophage subsets, Biochem. Pharmacol. source

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    Premisecitation

    S.a.-infected RAW cells showed increased matrix mtROS production; treatment with Necrox-5 reduced matrix mtROS levels and increased bacterial burden.

    “S.a.-infected RAW cells showed increased matrix mtROS production and treatment with Necrox-5, a rarely used mitochondrial matrix ROS scavenger, did not only reduce matrix mtROS levels but also increased bacterial burden indicating a role for matrix mtROS for defense against S.a.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [252] Abuaita, 2018, Mitochondria-derived vesicles deliver antimicrobial reactive oxygen species to control phagosome-localized staphylococcus aureus, Cell Host Microbe source

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    Premisecitation

    Mtb uses phagosomes of macrophages as a proliferative niche and inhibits fusion with lysosomes and avoids xenophagy.

    “Like S.a., Mycobacterium tuberculosis (Mtb) uses phagosomes of macrophages as proliferative niche. Mechanisms to survive in phagosomes are the inhibition of fusion with lysosomes and avoiding to be targeted by xenophagy [255].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [255] Queval, 2017, The macrophage: A disputed fortress in the battle against mycobacterium tuberculosis, Front. Microbiol. source

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    Premisecitation

    IRE1-deficient BMDM showed reduced bacterial burden and total cellular ROS levels, suggesting a ROS-mediated antibacterial role for IRE1.

    “Since IRE1-deficient BMDM showed reduced bacterial burden and total cellular ROS levels, a ROS-mediated antibacterial role for IRE1 was suggested.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [249] Abuaita, 2015, The endoplasmic reticulum stress sensor inositol-requiring enzyme 1alpha augments bacterial killing through sustained oxidant production, mBio source

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    Premisecitation

    Nox2-deficient mice showed increased bacterial burden in a pulmonary S.a. infection model, while Nox2-deficient BMDM in vitro did not differ in bacterial load.

    “An in vivo role for Nox2 during infection with S.a. was suggested in a pulmonary infection model [248]. While Nox2-deficent mice showed increased bacterial burden, Nox2-deficient BMDM in vitro did not differ in their bacterial load.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [248] Kohler, 2011, Nadph-oxidase but not inducible nitric oxide synthase contributes to resistance in a murine staphylococcus aureus newman pneumonia model, Microbes Infect. source

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    • cites [249] Abuaita, 2015, The endoplasmic reticulum stress sensor inositol-requiring enzyme 1alpha augments bacterial killing through sustained oxidant production, mBio source

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    Premisecitation

    Individuals with chronic granulomatous disease fail to produce sufficient amounts of ROS by Nox2 and suffer from increased susceptibility to infections.

    “Individuals with chronic granulomatous disease (CGD), a genetic disorder caused by hypo- or amorphic mutations in genes encoding for Nox2 or its subunits, fail to produce sufficient amounts of ROS by Nox2 and in consequence suffer from increased susceptibility to infections [236]”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [236] Yu, H.H., Yang, Y.H., and Chiang, B.L. (2020). Chronic granulomatous disease: A comprehensive review. Clin. Rev. Allergy Immunol. source

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    Premisecitation

    Macrophages often are the first immune cells that encounter invading pathogens.

    “Macrophages often are the first immune cells that encounter invading pathogens [219,220,221].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [219] Jung, 2018, Macrophages and monocytes in 2017: Macrophages and monocytes: Of tortoises and hares, Nat. Rev. Immunol. source

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    • cites [220] Epelman, 2014, Origin and functions of tissue macrophages, Immunity source

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    • cites [221] Davies, 2013, Tissue-resident macrophages, Nat. Immunol. source

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    Premisecitation

    Abuaita et al. suggested that TLR signaling induces delivery of mitochondria-derived vesicles containing SOD2 to S.a.-containing phagosomes, where SOD2 enhances conversion of O2− to H2O2.

    “Abuaita et al. suggested that TLR signaling, instead of triggering mtROS production, induces the delivery of mitochondria-derived vesicles containing the mitochondrial matrix protein SOD2 to S.a.-containing phagosomes. In the phagosomal lumen, the delivered SOD2 then enhances conversion of O2●− to H2O2.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [252] Abuaita, 2018, Mitochondria-derived vesicles deliver antimicrobial reactive oxygen species to control phagosome-localized staphylococcus aureus, Cell Host Microbe source

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    Premisecitation

    TLR2/4/9-deficient RAW cells infected with S.a. showed unaltered matrix mtROS production, while bacterial burden was reduced.

    “TLR-mediated signaling can induce mtROS production [30,61], but in contrast TLR2/4/9-deficient RAW cells infected with S.a. showed unaltered matrix mtROS production, while bacterial burden, however, was reduced.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [252] Abuaita, 2018, Mitochondria-derived vesicles deliver antimicrobial reactive oxygen species to control phagosome-localized staphylococcus aureus, Cell Host Microbe source

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    Premisecitation

    Early cytosolic mtROS released by opened mPTP led to killing of Mtb, but later resulted in necroptosis of macrophages.

    “The early effect of cytosolic mtROS released by the opened mPTP led to killing of Mtb, but later on it resulted in necroptosis of macrophages [129].”
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    • cites [129] Roca, 2013, Tnf dually mediates resistance and susceptibility to mycobacteria via mitochondrial reactive oxygen species, Cell source

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    Premisecitation

    Mtb infection alone was not sufficient to increase total cellular ROS levels or to induce matrix mtROS production in zebrafish macrophages.

    “Mtb infection alone was not sufficient to increase total cellular ROS levels or to induce matrix mtROS production in zebrafish macrophages.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [129] Roca, 2013, Tnf dually mediates resistance and susceptibility to mycobacteria via mitochondrial reactive oxygen species, Cell source

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    • cites [257] Roca, 2019, Tnf induces pathogenic programmed macrophage necrosis in tuberculosis through a mitochondrial-lysosomal-endoplasmic reticulum circuit, Cell source

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    Premisecitation

    Treatment of WT or Nox2-deficient BMDM with NAC or DPI resulted in reduced total cellular ROS levels, reduced bacterial load and reduced induction of IRE1.

    “However, treatment of WT or Nox2-deficient BMDM with the global ROS scavenger N-acetyl-cysteine (NAC) or the flavoprotein inhibitor DPI (see Section 6 and Section 7) resulted in reduced total cellular ROS levels, reduced bacterial load and reduced induction of the Inositol-Requiring Enzyme 1α (IRE1), which is a sensor of endoplasmatic reticulum stress.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [249] Abuaita, 2015, The endoplasmic reticulum stress sensor inositol-requiring enzyme 1alpha augments bacterial killing through sustained oxidant production, mBio source

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    Premisecitation

    LAP activation strictly requires Nox2-derived extracellular ROS production, but Nox2-derived ROS do not seem to be listericidal by themselves.

    “LAP activation strictly requires Nox2-derived extracellular ROS production, but notably, Nox2-derived ROS do not seem to be listericidal by themselves [58].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [58] Gluschko, 2018, The beta2 integrin mac-1 induces protective lc3-associated phagocytosis of listeria monocytogenes, Cell Host Microbe source

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    Premisecitation

    Mtb virulence factor CpsA actively inhibits Nox2 recruitment to phagosomes, preventing Nox2-derived ROS production and LAP killing.

    “Through its virulence factor CpsA, Mtb also actively inhibits Nox2 recruitment to phagosomes. By preventing Nox2-derived ROS production, Mtb effectively avoids being targeted and killed by LAP [241,256].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [241] Herb, 2020, Lc3-associated phagocytosis—The highway to hell for phagocytosed microbes, Semin. Cell Dev. Biol. source

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    • cites [256] Koster, 2017, Mycobacterium tuberculosis is protected from nadph oxidase and lc3-associated phagocytosis by the lcp protein cpsa, Proc. Natl. Acad. Sci. USA source

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    Premisecitation

    For L.m., the integrin Mac-1 is crucial for initiation of ROS production by Nox2, whereas TLR-mediated signaling is completely dispensable.

    “For L.m., the integrin Mac-1 is crucial for initiation of ROS production by Nox2, whereas Toll-like receptor (TLR)-mediated signaling is completely dispensable [58].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [58] Gluschko, 2018, The beta2 integrin mac-1 induces protective lc3-associated phagocytosis of listeria monocytogenes, Cell Host Microbe source

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    Premisecitation

    TNF priming induces production of antibacterial matrix mtROS by two converging signaling pathways: RIPK1-RIPK3-mediated calcium overload and mPTP opening via RIPK1, RIPK3 and MLKL.

    “Priming with TNF, however, induced production of antibacterial matrix mtROS by two converging signaling pathways. Firstly, TNF mediated induction of matrix mtROS production by ryanoid-receptor-receptor-interacting serine/threonine-protein kinase 1 (RIPK1)-RIPK3-mediated calcium overload of mitochondria [257]. Secondly, since matrix mtROS cannot escape the matrix of intact mitochondria [127,128,129”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [257] Roca, 2019, Tnf induces pathogenic programmed macrophage necrosis in tuberculosis through a mitochondrial-lysosomal-endoplasmic reticulum circuit, Cell source

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    • cites [129] Roca, 2013, Tnf dually mediates resistance and susceptibility to mycobacteria via mitochondrial reactive oxygen species, Cell source

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    Premisecitation

    MST1/2-deficient BMDM show reduced ROS production into phagosomes and increased bacterial burden during L.m. infection.

    “MST1/2-deficient BMDM showed reduced ROS production into phagosomes and increased bacterial burden.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [246] Geng, 2015, Kinases mst1 and mst2 positively regulate phagocytic induction of reactive oxygen species and bactericidal activity, Nat. Immunol. source

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    Premisecitation

    Recognition of bacteria by macrophages leads to ROS production in different cellular compartments, where they fulfill different antibacterial functions.

    “Recognition of bacteria by macrophages leads to ROS production in different cellular compartments, where they fulfill different antibacterial functions.”
    Quote in the paperSubstantial claim0 of 5 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

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    • cites [58] Gluschko, 2018, The beta2 integrin mac-1 induces protective lc3-associated phagocytosis of listeria monocytogenes, Cell Host Microbe source

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    • cites [233] Craig, M., and Slauch, J.M. (2009). Phagocytic superoxide specifically damages an extracytoplasmic target to inhibit or kill salmonella. PLoS ONE, 4. source

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    • cites [234] Wink, 2011, Nitric oxide and redox mechanisms in the immune response, J. Leukoc. Biol. source

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    • cites [235] Slauch, 2011, How does the oxidative burst of macrophages kill bacteria? Still an open question, Mol. Microbiol. source

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    Premisecitation

    Macrophages employ indirect antimicrobial mechanisms including activation of inflammasomes, secretion of cytokines and chemokines, and MHC-dependent presentation of pathogen-derived antigens.

    “Indirect antimicrobial mechanisms include the activation of inflammasomes and the secretion of cytokines and chemokines, which help to orchestrate the subsequent innate and adaptive immune responses [30,231], and the MHC-dependent presentation of pathogen-derived antigens [232].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [231] Descoteaux, 2014, Macrophage cytokines: Involvement in immunity and infectious diseases, Front. Immunol.

      Not checked in the free run.

    • cites [232] Allan, 2014, Nadph oxidase modifies patterns of mhc class ii-restricted epitopic repertoires through redox control of antigen processing, J. Immunol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Macrophages employ direct antimicrobial mechanisms including generation of RNS in the phagosome and delivery of cathepsins and other hydrolases into maturing phagosomes.

    “Beyond the production of ROS, macrophages also employ an array of directly antimicrobial mechanisms, e.g., the generation of reactive nitrogen species (RNS) in the phagosome and the delivery of cathepsins and other hydrolases into maturing phagosomes [59,225,226,227,228,229,230].”
    Quote in the paperNot a substantial claim0 of 7 sources checked
    • cites [59] Schramm, 2014, Riboflavin (vitamin b2) deficiency impairs nadph oxidase 2 (nox2) priming and defense against listeria monocytogenes, Eur. J. Immunol. source

      Not checked in the free run.

    • cites [225] Mitchell, G., Chen, C., and Portnoy, D.A. (2016). Strategies used by bacteria to grow in macrophages. Microbiol. Spectr., 4. source

      Not checked in the free run.

    • cites [226] Weiss, 2015, Macrophage defense mechanisms against intracellular bacteria, Immunol. Rev. source

      Not checked in the free run.

    • cites [227] Beck, 2006, Cutting edge: A novel nonoxidative phagosomal mechanism exerted by cathepsin-d controls listeria monocytogenes intracellular growth, J. Immunol. source

      Not checked in the free run.

    • cites [228] Aktan, 2004, Inos-mediated nitric oxide production and its regulation, Life Sci. source

      Not checked in the free run.

    • cites [229] Utermohlen, 2003, Severe impairment in early host defense against listeria monocytogenes in mice deficient in acid sphingomyelinase, J. Immunol. source

      Not checked in the free run.

    • cites [230] 2000, The contribution of both oxygen and nitrogen intermediates to the intracellular killing mechanisms of c1q-opsonized listeria monocytogenes by the macrophage-like ic-21 cell line, Immunology source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Macrophages engulf pathogens, dead cells and cellular debris by phagocytosis and subsequently degrade the cargo in phagolysosomes.

    “They engulf pathogens, dead cells and cellular debris by phagocytosis and subsequently degrade the cargo in phagolysosomes [222,223,224].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [222] Lemke, 2019, How macrophages deal with death, Nat. Rev. Immunol. source

      Not checked in the free run.

    • cites [223] Haas, 2007, The phagosome: Compartment with a license to kill, Traffic source

      Not checked in the free run.

    • cites [224] Djaldetti, 2002, Phagocytosis--the mighty weapon of the silent warriors, Microsc. Res. Tech. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Fe-SODB is conserved among Toxoplasma parasites, suggesting that other Toxoplasma strains may also possess the ability to detoxify phagosomal ROS.

    “Fe-SODB is conserved among Toxoplasma parasites suggesting that also other Toxoplasma strains may possess this ability, questioning the still controversially discussed direct antiparasitic role for ROS in T.g.-infected macrophages.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Leishmania amazonensis infection in J774 cells induces an increase in Nox2 protein levels and total cellular ROS levels.

    “Leishmania amazonensis infection in J774 cells induced an increase in Nox2 protein levels and total cellular ROS levels [290].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [290] Alonso, 2019, Hif-1alpha-regulated mif activation and nox2-dependent ros generation promote leishmania amazonensis killing by macrophages under hypoxia, Cell. Immunol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    E.c. infection of BMDM caused reduced complex I activity but enhanced complex II activity, and induced no matrix mtROS production.

    “In BMDM infected with E.c., mitochondria showed reduced complex I activity but enhanced complex II activity. While induction of matrix mtROS with rotenone [269], a commonly used positive control for matrix mtROS measurements (see Section 5.2 and Section 7) [30,270,271], was still possible, E.c. infection induced no matrix mtROS production suggesting that the activity shift to complex II did not”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [268] Garaude, 2016, Mitochondrial respiratory-chain adaptations in macrophages contribute to antibacterial host defense, Nat. Immunol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    S.t. infection of BMDM induced mitochondrial recruitment and matrix mtROS production near phagosomes in a TLR-MyD88-TRAF6-dependent manner, and matrix ROS contributed to inactivation of S.t.

    “They demonstrated for the first time that S.t. infection of BMDM induced mitochondrial recruitment and matrix mtROS production near phagosomes in a TLR-myeloid differentiation primary response 88 (MyD88)-TNF receptor associated factor 6 (TRAF6)-dependent manner. The matrix ROS production then contributed to the inactivation of S.t.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [61] West, 2011, Tlr signalling augments macrophage bactericidal activity through mitochondrial ros, Nature source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    ROS have to be produced in direct vicinity of the regulated target and do not excessively saturate the cell until they hit their target at random.

    “supporting the observation of other studies that ROS have to be produced in direct vicinity of the regulated target and do not excessively saturate the cell until they hit their target at random [29,30,37,61,125,273].”
    Quote in the paperSubstantial claim0 of 6 sources checked
    • cites [29] Sies, 2020, Reactive oxygen species (ros) as pleiotropic physiological signalling agents, Nat. Rev. Mol. Cell Biol. source

      Not checked in the free run.

    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [37] Beretta, 2020, Nox4 regulates insp3 receptor-dependent ca(2+) release into mitochondria to promote cell survival, EMBO J. source

      Not checked in the free run.

    • cites [61] West, 2011, Tlr signalling augments macrophage bactericidal activity through mitochondrial ros, Nature source

      Not checked in the free run.

    • cites [125] Pastukh, 2012, Perinuclear mitochondrial clustering creates an oxidant-rich nuclear domain required for hypoxia-induced transcription, Sci. Signal.

      Not checked in the free run.

    • cites [273] Zhou, 2011, A role for mitochondria in nlrp3 inflammasome activation, Nature source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    Several other Nox enzymes were suggested to be involved in defense against infection with protozoan parasites.

    “several other Nox enzymes were suggested to be involved in defense against infection with protozoan parasites.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    SIRT3 deficiency resulted in damaged mitochondria and high matrix mtROS, but these did not contribute to direct antibacterial defense; instead SIRT3 induced antibacterial xenophagy of Mtb.

    “Deficiency for SIRT3 resulted in mitochondria with swollen and disrupted cristae, and damaged mitochondria are known to produce high amounts of matrix mtROS [127,129,130,131]. However, these damage-induced matrix mtROS in SIRT3-deficicent BMDM were only a side effect and did not contribute to direct antibacterial defense. Instead, SIRT3 induced antibacterial xenophagy of Mtb that had escaped from”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [258] Kim, 2019, Sirt3 promotes antimycobacterial defenses by coordinating mitochondrial and autophagic functions, Autophagy source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox2-derived extracellular ROS production is independent of TLR signaling and depends on the integrin Mac-1, which is important for induction of the highly antimicrobial pathway of LAP.

    “Notably, Nox2-derived extracellular ROS production was completely independent of TLR signaling and instead depended on the integrin Mac-1, which was of crucial importance for induction of the highly antimicrobial pathway of LAP [58,241,243].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [58] Gluschko, 2018, The beta2 integrin mac-1 induces protective lc3-associated phagocytosis of listeria monocytogenes, Cell Host Microbe source

      Not checked in the free run.

    • cites [241] Herb, 2020, Lc3-associated phagocytosis—The highway to hell for phagocytosed microbes, Semin. Cell Dev. Biol. source

      Not checked in the free run.

    • cites [243] Herb, 2018, Lc3-associated phagocytosis initiated by integrin itgam-itgb2/mac-1 enhances immunity to listeria monocytogenes, Autophagy source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Salmonella typhimurium can evade degradation in the phagosome and replicate in macrophages.

    “Salmonella typhimurium (S.t.), a pathogen responsible for severe food-borne illness and a major cause of diarrheal diseases [259], also can evade degradation in the phagosome and replicate in macrophages [260].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [260] Eriksson, 2003, Unravelling the biology of macrophage infection by gene expression profiling of intracellular salmonella enterica, Mol. Microbiol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Toxoplasma parasites can detoxify phagosomal ROS, as demonstrated with Toxoplasma cruzii in phagosomes of J774A.1 macrophage-like cells and BMDM.

    “The ability of Toxoplasma parasites to detoxify phagosomal ROS was demonstrated with the evolutionary related Toxoplasma cruzii in phagosomes of J774A.1 macrophage-like cells (J774) and BMDM [283].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [283] Martinez, 2019, Cytosolic fe-superoxide dismutase safeguards trypanosoma cruzi from macrophage-derived superoxide radical, Proc. Natl. Acad. Sci. USA source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Infection of peritoneal macrophages with Listeria monocytogenes induces extracellular ROS production that is strictly Nox2-dependent and cytosolic ROS production that is completely independent of Nox2 and all other Nox isoforms.

    “Infection of PM with L.m. induced the production of extracellular ROS (see Section 5.2), which was strictly Nox2-dependent, and the production of cytosolic ROS (see Section 5.2), which was completely independent not only of Nox2 but also of all other Nox isoforms.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

  • May combine two claims.

    Premiseown data

    Toxoplasma gondii infection increases total cellular ROS levels in WT BMDM in vitro, and this increase is abolished in Nox1- or Nox2-deficient BMDM.

    “T.g. infection also increased total cellular ROS levels in WT BMDM in vitro, which was abolished in Nox1- or Nox2-deficient BMDM, while parasitic burden was increased, suggesting a role for Nox1 or Nox2 as important sources of ROS in antiparasitic defense.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Macrophages have a dual role in Leishmania infection: they kill some phagocytosed parasites but also constitute a replicative niche.

    “Macrophages manage to kill some of the phagocytozed parasites but also constitute a replicative niche [287,288].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [287] Scott, 2016, Cutaneous leishmaniasis: Immune responses in protection and pathogenesis, Nat. Rev. Immunol. source

      Not checked in the free run.

    • cites [288] Srivastava, 2016, Possibilities and challenges for developing a successful vaccine for leishmaniasis, Parasites Vectors source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Administration of the matrix mtROS scavenger mitoTEMPO to mice during influenza A virus infection decreases airway inflammation, body weight loss, and mortality.

    “Administration of the matrix mtROS scavenger mitoTEMPO to mice during infection with influenza A virus decreased inflammation of the airways, body weight loss and mortality.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [296] To, 2020, Mitochondrial reactive oxygen species contribute to pathological inflammation during influenza a virus infection in mice, Antioxid. Redox Signal. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Leishmania major actively impairs recruitment of Nox2 to phagosomes, leading to reduced extracellular ROS production and reduced activation of LAP.

    “At least one Leishmania subspecies, Leishmania major, actively impairs recruitment of Nox2 to phagosomes, which leads to reduced extracellular ROS production and reduced activation of LAP [241,289].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [241] Herb, 2020, Lc3-associated phagocytosis—The highway to hell for phagocytosed microbes, Semin. Cell Dev. Biol. source

      Not checked in the free run.

    • cites [289] Matte, C., Casgrain, P.A., Seguin, O., Moradin, N., Hong, W.J., and Descoteaux, A. (2016). Leishmania major promastigotes evade lc3-associated phagocytosis through the action of gp63. PLoS Pathog., 12. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox2-derived ROS inactivate phagosomal cathepsins L and S in a redox-dependent manner, reducing uncoordinated proteolysis of engulfed peptides and increasing proper antigen presentation by MHC class II molecules.

    “Nox2-dervied ROS inactivated in a redox-dependent manner the phagosomal cathepsins L and S, thereby reducing uncoordinated proteolysis of engulfed peptides and increasing proper presentation as antigens by major histocompatibility complex (MHC)-class II molecules (see Figure 6).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [232] Allan, 2014, Nadph oxidase modifies patterns of mhc class ii-restricted epitopic repertoires through redox control of antigen processing, J. Immunol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Phagocytosis of IgG-opsonized particles induces extracellular ROS production, which is abolished in Nox2-deficient BMDM.

    “Phagocytosis of immunoglobulin G (IgG)-opsonized particles induced extracellular ROS production, which was abolished in Nox2-deficient BMDM.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [232] Allan, 2014, Nadph oxidase modifies patterns of mhc class ii-restricted epitopic repertoires through redox control of antigen processing, J. Immunol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox1/Nox4 double-deficient mice resemble Nox1-deficient mice, excluding an interconnection between the two Nox enzymes.

    “Nox1/Nox4 double-deficient mice resembled Nox1-deficient mice excluding an interconnection between the two Nox enzymes.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [319] Liang, 2019, Nadph oxidase 1 in liver macrophages promotes inflammation and tumor development in mice, Gastroenterology source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox1-deficient, but not Nox4-deficient, animals show strongly reduced tumor numbers and size and reduced liver damage in a hepatocellular carcinoma model.

    “Nox1-deficient, but not Nox4-deficient, animals showed strongly reduced tumor numbers and size and reduced liver damage.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [319] Liang, 2019, Nadph oxidase 1 in liver macrophages promotes inflammation and tumor development in mice, Gastroenterology source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Treatment with MitoQ reduces only IL-1β cleavage, whereas TNF and IL-10 levels remain unaltered.

    “Moreover, treatment with MitoQ, a substance known to scavenge matrix mtROS (see Section 6) [314,315], gave contrasting results: only IL-1β cleavage was reduced, whereas TNF and IL-10 levels remained unaltered.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [314] Murphy, 2007, Targeting antioxidants to mitochondria by conjugation to lipophilic cations, Annu. Rev. Pharmacol. Toxicol. source

      Not checked in the free run.

    • cites [315] Kelso, 2001, Selective targeting of a redox-active ubiquinone to mitochondria within cells: Antioxidant and antiapoptotic properties, J. Biol. Chem. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    High concentrations of H2O2 (10 mM) disrupt the interaction of TXNIP with thioredoxin, leading to TXNIP interaction with NLRP3, resulting in caspase-1 activation and IL-1β cleavage.

    “High concentrations of H2O2 (10 mM) disrupted the interaction of the thioredoxin-interacting protein (TXNIP) with thioredoxin leading to its interaction with NLRP3 resulting in caspase-1 activation and finally IL-1β cleavage.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [334] Zhou, 2010, Thioredoxin-interacting protein links oxidative stress to inflammasome activation, Nat. Immunol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox1-deficient BMDM show no ROS production and no expression of cytokines.

    “Nox1-deficient BMDM showed no ROS production and no expression of cytokines, nicely corroborating the in vivo findings.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [319] Liang, 2019, Nadph oxidase 1 in liver macrophages promotes inflammation and tumor development in mice, Gastroenterology source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    After LPS stimulation, metformin-treated BMDM show reduced total cellular ROS levels, decreased IL-1β cleavage, decreased TNF secretion, and enhanced secretion of the anti-inflammatory cytokine IL-10.

    “After stimulation with LPS, metformin-treated BMDM showed reduced total cellular ROS levels (see Section 5.1), decreased IL-1β cleavage, decreased TNF secretion yet enhanced secretion of the anti-inflammatory cytokine IL-10, suggesting a regulating role for ROS during proinflammatory signaling, which can be inhibited via Metformin treatment.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [313] Kelly, 2015, Metformin inhibits the production of reactive oxygen species from nadh:Ubiquinone oxidoreductase to limit induction of interleukin-1beta (il-1beta) and boosts interleukin-10 (il-10) in lipopolysaccharide (lps)-activated macrophages, J. Biol. Chem. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox1-derived ROS production is necessary for upregulation of iNOS, but not vice versa.

    “Nox1-derived ROS production was necessary for upregulation of inducible NO synthase (iNOS), but not vice versa.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [320] Liu, 2020, Nox1/nadph oxidase in bone marrow-derived cells modulates intestinal barrier function, Free Radic. Biol. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    NLRP3-, Caspase-1-, and Caspase 1l-deficient macrophages show normal secretion of TNF and IL-6 after LPS stimulation, indicating that the NLRP3 inflammasome is not involved in TNF and IL-6 secretion.

    “Not surprisingly, the NLRP3 inflammasome was, with exception for IL-1β, not involved in cytokine secretion as NLRP3-, Caspase-1- and Caspase 1l-deficient macrophages showed normal secretion of TNF and IL-6 (see Section 4.3).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [63] Bulua, 2011, Mitochondrial reactive oxygen species promote production of proinflammatory cytokines and are elevated in tnfr1-associated periodic syndrome (traps), J. Exp. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Macrophages lacking the p22phox subunit show cytokine secretion similar to Nox2-deficient macrophages, indicating that Nox enzymes are dispensable for cytokine secretion after LPS stimulation.

    “Moreover, macrophages lacking the p22phox subunit showed cytokine secretion similar to Nox2-deficient macrophages, nicely showing that Nox enzymes were dispensable for cytokine secretion after LPS stimulation.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [63] Bulua, 2011, Mitochondrial reactive oxygen species promote production of proinflammatory cytokines and are elevated in tnfr1-associated periodic syndrome (traps), J. Exp. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox2 deficiency in antigen-presenting BMDM results in increased cleavage of several investigated phagocytosed antigens and reduced MHC class II-dependent presentation.

    “Accordingly, Nox2 deficiency of antigen-presenting BMDM resulted in increased cleavage of several investigated phagocytosed antigens and reduced MHC-class II-dependent presentation.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [232] Allan, 2014, Nadph oxidase modifies patterns of mhc class ii-restricted epitopic repertoires through redox control of antigen processing, J. Immunol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Cytokine secretion is not altered in Nox2-deficient or Nox2/TNFR1 double-deficient macrophages.

    “Cytokine secretion was not altered in Nox2-deficient or Nox2/TNFR1 double-deficient macrophages.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [63] Bulua, 2011, Mitochondrial reactive oxygen species promote production of proinflammatory cytokines and are elevated in tnfr1-associated periodic syndrome (traps), J. Exp. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In Nox1-deficient animals after chemically induced liver damage, total ROS levels in tumor tissue, tumor cell proliferation, and hepatic levels of IL-6 and TNF are reduced.

    “Total ROS levels in tumor tissue, tumor cell proliferation and hepatic levels of the proinflammatory cytokines IL-6 and TNF were reduced after chemically induced liver damage in Nox1-deficient animals.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [319] Liang, 2019, Nadph oxidase 1 in liver macrophages promotes inflammation and tumor development in mice, Gastroenterology source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox1-deficient ileal macrophages, but not epithelial cells, show reduced iNOS expression after LPS stimulation.

    “Nox1-deficient ileal macrophages, but not epithelial cells, showed reduced iNOS expression after LPS stimulation.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [320] Liu, 2020, Nox1/nadph oxidase in bone marrow-derived cells modulates intestinal barrier function, Free Radic. Biol. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In both Nox1- and iNOS-deficient animals, the intestinal barrier is impaired, whereas cell death and inflammation in the ileum are unaffected.

    “In both Nox1-and iNOS-deficient animals, the intestinal barrier was impaired whereas cell death and inflammation in the ileum were unaffected.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [320] Liu, 2020, Nox1/nadph oxidase in bone marrow-derived cells modulates intestinal barrier function, Free Radic. Biol. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Liver macrophages, identified via cell-type specific Nox1 deletion, promote a proinflammatory liver environment, hepatocyte proliferation, tumor development, and liver damage, while Nox1 deletion in hepatocytes and bile duct cells has no effect.

    “Liver macrophages were identified via cell-type specific Nox1 deletion as promoters of a proinflammatory liver environment, hepatocyte proliferation, tumor development and liver damage, while Nox1 deletion in hepatocytes and bile duct cells had no effect.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [319] Liang, 2019, Nadph oxidase 1 in liver macrophages promotes inflammation and tumor development in mice, Gastroenterology source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Recognition of DAMPs released from damaged hepatocytes increases total cellular ROS levels and induces ERK1/2-mediated expression of IL-6 and TNF in BMDM.

    “Mechanistically, recognition of damage-associated molecular patterns (DAMP) released from damaged hepatocytes in vitro increased total cellular ROS levels (see also Section 5.1) and induced extracellular signal-regulated kinases 1/2 (ERK1/2)-mediated expression of IL-6 and TNF in BMDM.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [319] Liang, 2019, Nadph oxidase 1 in liver macrophages promotes inflammation and tumor development in mice, Gastroenterology source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Expression of a redox-insensitive NEMO mutant in peritoneal macrophages, BMDM, and MEF completely abolishes disulfide linkage of NEMO, proinflammatory signaling, and cytokine secretion in response to bacterial infection.

    “expression of a redox-insensitive NEMO mutant in PM, BMDM and MEF through transfection of in vitro-generated mRNA [123] completely abolished disulfide linkage of NEMO, proinflammatory signaling and cytokine secretion in response to bacterial infection.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [123] Herb, 2019, Highly efficient transfection of primary macrophages with in vitro transcribed mrna, J. Vis. Exp.

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    PMN recruitment to the peritoneal cavity is similar in WT and Nox2-deficient animals, but Nox2-deficient animals show prolonged PMN persistence and enhanced chemokine secretion.

    “Polymorphnuclear leukocyte (PMN) recruitment to the peritoneal cavity was similar in WT and Nox2-deficient animals, however, the latter showed prolonged persistence of PMN and the recruited cells showed enhanced secretion of chemokines.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [318] Whitmore, 2013, Nox2 protects against prolonged inflammation, lung injury, and mortality following systemic insults, J. Innate Immun. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox1-deficient animals show reduced cleavage of tight junctions in the intestine, suggesting a critical role for Nox1-derived ROS in regulation of iNOS expression and MMP9 secretion by intestinal macrophages and subsequent cleavage of ileal tight junctions.

    “Accordingly, Nox1-deficient animals showed reduced cleavage of tight junctions in the intestine suggesting a critical role for Nox1-derived ROS in regulation of iNOS expression and MMP9 secretion by intestinal macrophages and subsequent cleavage of ileal tight junctions.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [320] Liu, 2020, Nox1/nadph oxidase in bone marrow-derived cells modulates intestinal barrier function, Free Radic. Biol. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Phagocytosis of uric acid crystals leads to an increase in total cellular ROS levels and dissociation of TXNIP from thioredoxin.

    “Phagocytosis of uric acid crystals, as inflammasome-activating stimulus, also led to increase in total cellular ROS levels (see also Section 5.1) and dissociation of TXNIP from thioredoxin.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [334] Zhou, 2010, Thioredoxin-interacting protein links oxidative stress to inflammasome activation, Nat. Immunol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox2-deficient animals show increased PMN recruitment to the lung, more thrombi, and hemorrhage.

    “Bronchoalveolar lavage analysis revealed also increased recruitment of PMN to the lung in Nox2-deficient animals as well as more thrombi and hemorrhage.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [318] Whitmore, 2013, Nox2 protects against prolonged inflammation, lung injury, and mortality following systemic insults, J. Innate Immun. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    WT BMDM treated with LPS/ATP show enhanced expression of Nox4, whereas Nox4-deficient BMDM treated with ATP/LPS show reduced levels of IL-1β and reduced ASC speck formation.

    “WT BMDM treated with LPS/ATP showed enhanced expression of Nox4, whereas Nox4-deficient BMDM treated with ATP/LPS showed reduced levels of IL-1β and reduced ASC speck formation.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [343] Moon, 2016, Nox4-dependent fatty acid oxidation promotes nlrp3 inflammasome activation in macrophages, Nat. Med. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    For analysis of compartment-specific functions of ROS, the use of other substances is required.

    “For analysis of compartment-specific functions of ROS, the use of other substances is required.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Isoluminol is cell-impermeable and ideal for exclusive measurement of extracellular ROS, including ROS produced into endosomes and phagosomes; it cannot differentiate ROS subspecies.

    “Isoluminol is a derivative of the chemiluminescent luminol with the amino group placed on a more exposed position on the molecule. This modification makes isoluminol cell-impermeable. This makes it ideal for exclusive measurement of extracellular ROS, which also includes ROS produced into the lumen of endosomes and phagosomes [30,58,99,254,353,361]. However, the probe cannot differ between”
    Quote in the paperNot a substantial claim0 of 6 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [58] Gluschko, 2018, The beta2 integrin mac-1 induces protective lc3-associated phagocytosis of listeria monocytogenes, Cell Host Microbe source

      Not checked in the free run.

    • cites [99] Wolf, 2020, The tspo-nox1 axis controls phagocyte-triggered pathological angiogenesis in the eye, Nat. Commun. source

      Not checked in the free run.

    • cites [254] Lundqvist, 1996, Isoluminol-enhanced chemiluminescence: A sensitive method to study the release of superoxide anion from human neutrophils, Free Radic. Biol. Med. source

      Not checked in the free run.

    • cites [353] Walrand, 2002, Is the neutrophil reactive oxygen species production measured by luminol and lucigenin chemiluminescence intra or extracellular? Comparison with dcfh-da flow cytometry and cytochrome c reduction, Clin. Chim. Acta Int. J. Clin. Chem. source

      Not checked in the free run.

    • cites [361] Dahlgren, 1999, Respiratory burst in human neutrophils, J. Immunol. Methods source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Trolox, like Tiron, is a water-soluble derivative of vitamin E and globally scavenges ●OOH and ●OOR.

    “Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), like Tiron, is a water-soluble derivative of vitamin E and globally scavenges ●OOH and ●OOR [378,379].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [378] Dugas, 2000, Evaluation of the total peroxyl radical-scavenging capacity of flavonoids: Structure-activity relationships, J. Nat. Prod. source

      Not checked in the free run.

    • cites [379] Davies, 1988, Vitamin e analogue trolox c. E.S.R. And pulse-radiolysis studies of free-radical reactions, Biochem. J.

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    As a globally working ROS scavenger, usage of NAC can give only insights into general involvement of ROS in the process of interest.

    “As a globally working ROS scavenger, usage of NAC can give only insights into general involvement of ROS in the process of interest.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: NAC is a commonly used global ROS scavenger that works extra- and intracellularly.

  • May combine two claims.

    Premisereasoning

    5/6-Carboxy-DCF is retained in the cytosol and can specifically detect cytosolic ROS levels, but cannot differentiate ROS subspecies.

    “5-(and -6)-Carboxy-2′,7′-dihydrochlorofluorescein-diacetat (5/6-Carboxy-DCF) is a derivative of H2DCF-DA that contains two additional carboxyl groups that enhance its hydrophilicity and therefore strongly increase its retention in the cytosol. Therefore, in contrast to H2DCF-DA, which detects total cellular ROS levels, the green fluorescent 5/6-Carboxy-DCF can be used to specifically detect cytos”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [99] Wolf, 2020, The tspo-nox1 axis controls phagocyte-triggered pathological angiogenesis in the eye, Nat. Commun. source

      Not checked in the free run.

    • cites [359] Hempel, 1999, Dihydrofluorescein diacetate is superior for detecting intracellular oxidants: Comparison with 2′,7′-dichlorodihydrofluorescein diacetate, 5(and 6)-carboxy-2′,7′-dichlorodihydrofluorescein diacetate, and dihydrorhodamine 123, Free Radic. Biol. Med. source

      Not checked in the free run.

    • cites [364] Mak, 2017, Glutathione primes t cell metabolism for inflammation, Immunity source

      Not checked in the free run.

  • May combine two claims.

    Premisereasoning

    H2DCF-DA is cell-permeable, its acetate groups are cleaved by cellular esterases to H2DCF, which can be oxidized by ROS to fluorescent DCF; because it remains diffusible, only total cellular ROS levels can be detected.

    “2′,7′-Dichlordihydrofluorescein-diacetat (H2DCF-DA) is a cell-permeable derivative of fluorescein and one of the most commonly used ROS probes. After its diffusion into the cytosol, the two acetate groups are cleaved by cellular esterases generating 2′,7′-dichlordihydrofluorescein (H2DCF). This molecule can now be oxidized by ROS to generate the highly green fluorescent 2′,7′-dichlorofluorescein (”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [355] Ushijima, 1997, Chemiluminescence from human polymorphonuclear leukocytes activated with opsonized zymosan, Free Radic. Biol. Med. source

      Not checked in the free run.

    • cites [359] Hempel, 1999, Dihydrofluorescein diacetate is superior for detecting intracellular oxidants: Comparison with 2′,7′-dichlorodihydrofluorescein diacetate, 5(and 6)-carboxy-2′,7′-dichlorodihydrofluorescein diacetate, and dihydrorhodamine 123, Free Radic. Biol. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Amplex Red is cell-impermeable and specifically detects H2O2; in the presence of HRP and H2O2, it is oxidized to red fluorescent 10-acetyl-3,7-dihydroxypenoxazine, making it optimal for extracellular H2O2.

    “Amplex Red also is cell-impermeable but, in contrast to isoluminol, detects specifically H2O2 [362]. In the presence of HRP and H2O2, Amplex Red is oxidized to red fluorescent 10-acetyl-3,7-dihydroxypenoxazine [363]. In consequence, this probe is optimal for determination of specifically extracellular generation of H2O2.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [362] Karakuzu, 2019, Amplex red assay for measuring hydrogen peroxide production from caenorhabditis elegans, Bio Protoc. source

      Not checked in the free run.

    • cites [363] Mohanty, 1997, A highly sensitive fluorescent micro-assay of h2o2 release from activated human leukocytes using a dihydroxyphenoxazine derivative, J. Immunol. Methods source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Location of Nox4 to mitochondria was demonstrated by Western blot analysis of cellular fractions, and Nox4-deficient BMDM show reduced matrix mtROS production after stimulation with LPS and nigericin.

    “Location of Nox4 to mitochondria was demonstrated by Western blot analysis of cellular fractions and Nox4-deficient BMDM showed reduced matrix mtROS production after stimulation with LPS and nigericin.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [343] Moon, 2016, Nox4-dependent fatty acid oxidation promotes nlrp3 inflammasome activation in macrophages, Nat. Med. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Tiron is a vitamin E analog, cell-permeable and acts as a global O2●− scavenger.

    “Tiron (1,2-dihydroxybenzene-3,5-disulfonate) is a vitamin E analog, cell-permeable and acts as a global O2●− scavenger [375,376,377].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [375] Manzano, 2000, Human renal mesangial cells are a target for the anti-inflammatory action of 9-cis retinoic acid, Br. J. Pharm. source

      Not checked in the free run.

    • cites [376] Krishna, 1992, The catecholic metal sequestering agent 1,2-dihydroxybenzene-3,5-disulfonate confers protection against oxidative cell damage, Arch. Biochem. Biophys. source

      Not checked in the free run.

    • cites [377] Hein, 1998, Ldls impair vasomotor function of the coronary microcirculation: Role of superoxide anions, Circ. Res. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Lucigenin does not directly interact with O2●− but instead with cytochrome c and XO, increases NADPH oxidation, and binds NO with low affinity; therefore it should not be used for ROS detection.

    “However, other studies demonstrated that this probe does not directly interact with O2●−, but instead with cytochrome c and XO [357], increases NADPH oxidation and binds to NO with low affinity [358]. Since a big repertoire of other ROS probes is available, lucigenin should therefore not be used for ROS detection.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [357] Ostrachovitch, 1999, Lucigenin is a mediator of cytochrome c reduction but not of superoxide production, Arch. Biochem. Biophys. source

      Not checked in the free run.

    • cites [358] Skatchkov, 1999, Validation of lucigenin as a chemiluminescent probe to monitor vascular superoxide as well as basal vascular nitric oxide production, Biochem. Biophys. Res. Commun. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    MitoSOX accumulates in the mitochondrial matrix and specifically detects O2●−, but can be oxidized by other ROS while diffusing into mitochondria; Nox2 can cause false positive signals.

    “MitoSOX Red (MitoSOX) is a modified DHE analog attached to a triphenylphosphonium (TPP) group, which leads to accumulation of the probe in the mitochondrial matrix [365,366]. Like DHE itself, MitoSOX specifically detects O2●− and emits red fluorescence after the reaction (see Figure 7) [30,99,360]. In contrast to DHE, MitoSOX does so mainly in the mitochondrial matrix, though. Notably, MitoSOX can”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [365] Mukhopadhyay, 2007, Simultaneous detection of apoptosis and mitochondrial superoxide production in live cells by flow cytometry and confocal microscopy, Nat. Protoc. source

      Not checked in the free run.

    • cites [366] Robinson, 2006, Selective fluorescent imaging of superoxide in vivo using ethidium-based probes, Proc. Natl. Acad. Sci. USA source

      Not checked in the free run.

    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [99] Wolf, 2020, The tspo-nox1 axis controls phagocyte-triggered pathological angiogenesis in the eye, Nat. Commun. source

      Not checked in the free run.

    • cites [360] Wang, 2018, Measurement of reactive oxygen species (ros) and mitochondrial ros in ampk knockout mice blood vessels, Methods Mol. Biol source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    DHE reacts rather specifically with cellular O2●−, producing ethidium and 2-hydroxyethidium; it crosses membranes and can be oxidized anywhere, so only total cellular O2●− production can be detected; DNA binding can cause false localization interpretations.

    “Dihydroethidium (DHE) is a red fluorescent probe that rather specifically reacts with cellular O2●−, resulting in the two fluorescent products ethidium and 2-hydroxyethidium. Only 2-hydroxyethidium is generated after reaction with O2●−, while ethidium is formed by nonspecific redox reactions. However, DHE easily crosses cellular membranes and therefore can be oxidized by O2●− anywhere in and”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [360] Wang, 2018, Measurement of reactive oxygen species (ros) and mitochondrial ros in ampk knockout mice blood vessels, Methods Mol. Biol source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Ebselen is a heterocyclic seleno-organic compound and exerts glutathione reductase activity.

    “Ebselen is a heterocyclic seleno-organic compound and exerts glutathione reductase activity.”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [380] Matsushita, 2004, Effect of ebselen, a scavenger of reactive oxygen species, on chondrocyte metabolism, Mod. Rheumatol. source

      Not checked in the free run.

    • cites [381] Mugesh, 2013, Glutathione peroxidase activity of ebselen and its analogues: Some insights into the complex chemical mechanisms underlying the antioxidant activity, Curr. Chem. Biol. source

      Not checked in the free run.

    • cites [382] Nakamura, 2002, Ebselen, a glutathione peroxidase mimetic seleno-organic compound, as a multifunctional antioxidant. Implication for inflammation-associated carcinogenesis, J. Biol. Chem. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Tempol is a SOD mimetic and catalyzes the dismutation of O2●− into H2O2.

    “Tempol (4-Hydroxy-Tempo) is a SOD mimetic and catalyzes the dismutation of O2●− into H2O2.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [372] Bernardy, 2017, Tempol, a superoxide dismutase mimetic agent, inhibits superoxide anion-induced inflammatory pain in mice, Biomed. Res. Int. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nox4-deficient BMDM show reduced fatty acid uptake into mitochondria, reduced fatty acid oxidation, and reduced levels of CPT1.

    “Nox4-deficient BMDM also showed reduced fatty acid uptake into mitochondria, reduced fatty acid oxidation and reduced levels of carnitine palmitoyltransferase 1 (CPT1), a key enzyme for fatty acid oxidation.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [343] Moon, 2016, Nox4-dependent fatty acid oxidation promotes nlrp3 inflammasome activation in macrophages, Nat. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Global scavenging of ROS with NAC or matrix mtROS scavenging with mitoTEMPO reduces Caspase-1 activation and IL-1β secretion, while Nox2-deficiency has no effect.

    “Global scavenging of ROS with NAC or matrix mtROS scavenging with mitoTEMPO resulted in reduced Caspase-1 activation and IL-1β secretion, while Nox2-deficiency had no effect.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [346] Crane, 2014, Mitochondrial ros potentiates indirect activation of the aim2 inflammasome, Front. Microbiol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Tempol is cell-permeable and not compartment-specific and hence reduces O2●− levels everywhere in the cell.

    “Of note, Tempol is cell-permeable and not compartment-specific and hence reduces O2●− levels everywhere in the cell [373,374].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [373] Rak, 2000, Neuroprotection by the stable nitroxide tempol during reperfusion in a rat model of transient focal ischemia, J. Neurosurg. source

      Not checked in the free run.

    • cites [374] Nassar, 1999, Effects of the superoxide dismutase-mimic compound tempol on oxidant stress-mediated endothelial dysfunction, Antioxid. Redox Signal. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    GKT136901 and GKT137831 inhibit Nox1, Nox4 and Nox5 and in higher concentrations also Nox2, Duox1, Duox2, and XO.

    “Both compounds inhibit Nox1, Nox4 and Nox5 and in higher concentrations also Nox2, Duox1, Duox2 [406,407,408,409], and XO [392,393,408,409].”
    Quote in the paperNot a substantial claim0 of 6 sources checked
    • cites [406] Strengert, 2014, Mucosal reactive oxygen species are required for antiviral response: Role of duox in influenza a virus infection, Antioxid. Redox Signal. source

      Not checked in the free run.

    • cites [407] Sedeek, 2010, Critical role of nox4-based nadph oxidase in glucose-induced oxidative stress in the kidney: Implications in type 2 diabetic nephropathy, Am. J. Physiol. Ren. Physiol. source

      Not checked in the free run.

    • cites [408] Gaggini, 2011, Design, synthesis and biological activity of original pyrazolo-pyrido-diazepine, -pyrazine and -oxazine dione derivatives as novel dual nox4/nox1 inhibitors, Bioorgan. Med. Chem. source

      Not checked in the free run.

    • cites [409] Aoyama, 2012, Nicotinamide adenine dinucleotide phosphate oxidase in experimental liver fibrosis: Gkt137831 as a novel potential therapeutic agent, Hepatology source

      Not checked in the free run.

    • cites [392] Teixeira, 2017, Therapeutic potential of nadph oxidase 1/4 inhibitors, Br. J. Pharm. source

      Not checked in the free run.

    • cites [393] Altenhofer, 2015, Evolution of nadph oxidase inhibitors: Selectivity and mechanisms for target engagement, Antioxid. Redox Signal. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Vitamin C acts as a water-soluble reducing agent, donating electrons to ROS and therefore acts as a ROS eliminating substance.

    “Vitamin C (ascorbic acid and its anion ascorbate) acts as water-soluble reducing agent [383], donating electrons to ROS and therefore acts as ROS eliminating substance [384,385].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [383] Frei, 1989, Ascorbate is an outstanding antioxidant in human blood plasma, Proc. Natl. Acad. Sci. USA source

      Not checked in the free run.

    • cites [384] Retsky, 1993, Ascorbic acid oxidation product(s) protect human low density lipoprotein against atherogenic modification. Anti- rather than prooxidant activity of vitamin c in the presence of transition metal ions, J. Biol. Chem. source

      Not checked in the free run.

    • cites [385] Du, 2012, Ascorbic acid: Chemistry, biology and the treatment of cancer, Biochim. Biophys. Acta

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    VAS2870 is a well validated NADPH oxidase inhibitor as it shows no intrinsic antioxidant activity, does not inhibit other flavoproteins and inhibits NADPH oxidase-mediated ROS production in vitro and in vivo.

    “VAS2870 is a well validated NADPH oxidase inhibitor [341] as it shows no intrinsic antioxidant activity, does not inhibit other flavoproteins and inhibits NADPH oxidase-mediated ROS production in vitro and in vivo.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [341] Altenhofer, 2012, The nox toolbox: Validating the role of nadph oxidases in physiology and disease, Cell Mol. Life Sci. source

      Not checked in the free run.

  • May combine two claims.

    Premisenone given

    ROS scavengers directly remove ROS from the system, but optimally do not influence the sources of ROS production.

    “While ROS scavengers directly remove ROS from the system, but optimally do not influence the sources of ROS production”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Ebselen effectively removes H2O2 and ONOO−.

    “It effectively removes H2O2 and ONOO− [30,380,381,382].”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [380] Matsushita, 2004, Effect of ebselen, a scavenger of reactive oxygen species, on chondrocyte metabolism, Mod. Rheumatol. source

      Not checked in the free run.

    • cites [381] Mugesh, 2013, Glutathione peroxidase activity of ebselen and its analogues: Some insights into the complex chemical mechanisms underlying the antioxidant activity, Curr. Chem. Biol. source

      Not checked in the free run.

    • cites [382] Nakamura, 2002, Ebselen, a glutathione peroxidase mimetic seleno-organic compound, as a multifunctional antioxidant. Implication for inflammation-associated carcinogenesis, J. Biol. Chem. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    DPI's definition as a specific Nox2 inhibitor is outdated and statements about Nox2 involvement based on sole use of DPI are highly questionable.

    “Therefore, its definition as specific Nox2 inhibitor is outdated and statements about Nox2 involvement based on sole use of this substance are highly questionable.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Apocynin was long considered as a specific Nox2 inhibitor, but several studies have shown that this is not the case.

    “Apocynin was long considered as specific Nox2 inhibitor. However, several studies have shown that this is indeed not the case.”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [340] Wingler, 2011, Nox1, 2, 4, 5: Counting out oxidative stress, Br. J. Pharm. source

      Not checked in the free run.

    • cites [395] Trevelin, 2016, Apocynin and nox2 regulate nf-kappab by modifying thioredoxin-1 redox-state, Sci. Rep. source

      Not checked in the free run.

    • cites [396] Heumuller, 2008, Apocynin is not an inhibitor of vascular nadph oxidases but an antioxidant, Hypertension source

      Not checked in the free run.

    • cites [397] Aldieri, 2008, Classical inhibitors of nox nad(p)h oxidases are not specific, Curr. Drug Metab. source

      Not checked in the free run.

    • cites [398] Weiwer, 2009, Inhibition of human vascular nadph oxidase by apocynin derived oligophenols, Bioorgan. Med. Chem. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    GKT136901 also scavenges ONOO−, a RNS, and therefore works more like an antioxidant than a specific Nox inhibitor.

    “Notably, GKT136901 also scavenges ONOO−, a RNS [410] and therefore works more like an antioxidant than a specific Nox inhibitor.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [410] Schildknecht, 2014, The nox1/4 inhibitor gkt136901 as selective and direct scavenger of peroxynitrite, Curr. Med. Chem. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Interpretationreasoning

    For compartment-specific removal of ROS, substances described below are required.

    “For this, compartment-specific removal of ROS is required, which is possible with the substances described below.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    MitoTEMPO is a combination of antioxidant TEMPO attached to the TPP group and hence dismutates O2●− to H2O2 in the mitochondrial matrix.

    “MitoTEMPO is a combination of the antioxidant TEMPO attached to the TPP group and hence dismutates O2●− to H2O2 in the mitochondrial matrix [314,391]”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [314] Murphy, 2007, Targeting antioxidants to mitochondria by conjugation to lipophilic cations, Annu. Rev. Pharmacol. Toxicol. source

      Not checked in the free run.

    • cites [391] Smith, 1999, Selective targeting of an antioxidant to mitochondria, Eur. J. Biochem. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    The use of ROS source inhibitors is an option to block ROS production and analyze possible ROS sources when genetic modifications are not an option.

    “the use of ROS source inhibitors is an option to block ROS production and analyze possible ROS sources when genetic modifications are not an option.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    GKT136901 and GKT137831 were identified by Laleul and colleagues during a screen for Nox4 inhibitors.

    “GKT136901 and GKT137831 were identified by Laleul and colleagues during a screen for Nox4 inhibitors [405].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [405] Laleu, 2010, First in class, potent, and orally bioavailable nadph oxidase isoform 4 (nox4) inhibitors for the treatment of idiopathic pulmonary fibrosis, J. Med. Chem. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation

    No direct antioxidative side effects of GKT137831 have been reported so far, highlighting it as the most promising inhibitor for Nox enzymes in vitro and in vivo.

    “In contrast, so far no direct antioxidative side effects of GKT 137831 were reported [392] highlighting this substance as the most promising inhibitor for Nox enzymes not only in vitro but also in vivo [94,409,411,412,413,414].”
    Quote in the paperNot a substantial claim0 of 7 sources checked
    • cites [392] Teixeira, 2017, Therapeutic potential of nadph oxidase 1/4 inhibitors, Br. J. Pharm. source

      Not checked in the free run.

    • cites [94] Carnesecchi, 2011, A key role for nox4 in epithelial cell death during development of lung fibrosis, Antioxid. Redox Signal. source

      Not checked in the free run.

    • cites [409] Aoyama, 2012, Nicotinamide adenine dinucleotide phosphate oxidase in experimental liver fibrosis: Gkt137831 as a novel potential therapeutic agent, Hepatology source

      Not checked in the free run.

    • cites [411] Gorin, 2015, Targeting nadph oxidase with a novel dual nox1/nox4 inhibitor attenuates renal pathology in type 1 diabetes, Am. J. Physiol. Ren. Physiol. source

      Not checked in the free run.

    • cites [412] Carnesecchi, 2009, Nadph oxidase-1 plays a crucial role in hyperoxia-induced acute lung injury in mice, Am. J. Respir. Crit. Care Med. source

      Not checked in the free run.

    • cites [413] Green, 2012, The nox4 inhibitor gkt137831 attenuates hypoxia-induced pulmonary vascular cell proliferation, Am. J. Respir. Cell Mol. Biol. source

      Not checked in the free run.

    • cites [414] Bettaieb, 2015, Hepatocyte nicotinamide adenine dinucleotide phosphate reduced oxidase 4 regulates stress signaling, fibrosis, and insulin sensitivity during development of steatohepatitis in mice, Gastroenterology source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    DPI inhibits Nox enzymes, complex I of the ETC, iNOS, XO, and calcium transporters.

    “This includes Nox enzymes but also, for example, complex I of the ETC [63,250,251], iNOS [400,401] and XO [394,399] as well as calcium transporters [402].”
    Quote in the paperNot a substantial claim0 of 7 sources checked
    • cites [63] Bulua, 2011, Mitochondrial reactive oxygen species promote production of proinflammatory cytokines and are elevated in tnfr1-associated periodic syndrome (traps), J. Exp. Med. source

      Not checked in the free run.

    • cites [250] Lambeth, 2008, Nox enzymes as novel targets for drug development, Semin. Immunopathol. source

      Not checked in the free run.

    • cites [251] Bloxham, 1979, The relationship of diphenyleneiodonium-induced hypoglycaemia to the specific covalent modification of nadh-ubiquinone oxidoreductase, Biochem. Soc. Trans. source

      Not checked in the free run.

    • cites [400] Geyer, 1997, Nitric oxide synthase activity in tissues of the bovine eye, Graefe. Arch. Clin. Exp. Ophthalmol. Albrecht von Graefes Arch. Klin. Exp. Ophthalmol. source

      Not checked in the free run.

    • cites [401] Stuehr, 1991, Inhibition of macrophage and endothelial cell nitric oxide synthase by diphenyleneiodonium and its analogs, FASEB J. Off. Publ. Fed. Am. Soc. Exp. Biol.

      Not checked in the free run.

    • cites [394] Wind, 2010, Comparative pharmacology of chemically distinct nadph oxidase inhibitors, Br. J. Pharm. source

      Not checked in the free run.

    • cites [399] Tew, 1993, Studies on the inhibitory mechanism of iodonium compounds with special reference to neutrophil nadph oxidase, Biochem. J.

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Vitamin E does not scavenge nonradical ROS like H2O2 and does not scavenge ROS in hydrophilic cellular compartments such as the lumen of cell organelles or the cytosol.

    “It does not scavenge nonradical ROS like H2O2 and does not scavenge ROS in hydrophilic cellular compartments such as the lumen of cell organelles or the cytosol [75].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [75] Niki, 2016, Oxidative stress and antioxidants: Distress or eustress?, Arch. Biochem. Biophys. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    VAS2870 was the first published inhibitor that resulted from a systematic screening effort for selective NADPH oxidase inhibitors by Tegtmeier and colleagues.

    “VAS2870 was the first published inhibitor that resulted from a systematic screening effort for selective NADPH oxidase inhibitors by Tegtmeier and colleagues [394].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [394] Wind, 2010, Comparative pharmacology of chemically distinct nadph oxidase inhibitors, Br. J. Pharm. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Vitamin E is a lipophilic molecule that is exclusively integrated into biological membranes, where it efficiently scavenges radical ROS, mainly ●OOR and therefore acts as a major defense against lipid peroxidation in cells.

    “Vitamin E is a lipophilic molecule that is exclusively integrated into biological membranes, where it efficiently scavenges radical ROS, mainly ●OOR and therefore acts a major defense against lipid peroxidation in cells [388,389,390].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [388] Grabowski, 2005, Tocopherol content and activities of tyrosine aminotransferase and cystine lyase in arabidopsis under stress conditions, J. Plant Physiol. source

      Not checked in the free run.

    • cites [389] Ni, 2012, Vitamin e protects against lipid peroxidation and rescues tumorigenic phenotypes in cowden/cowden-like patient-derived lymphoblast cells with germline sdhx variants, Clin. Cancer Res. source

      Not checked in the free run.

    • cites [390] Newaz, 1998, Effect of alpha-tocopherol on lipid peroxidation and total antioxidant status in spontaneously hypertensive rats, Am. J. Hypertens. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    MitoTEMPO and MitoQ both are targeted to and accumulated in mitochondria due to their TPP group.

    “MitoTEMPO and MitoQ both are targeted to and accumulated in mitochondria due to their TPP group.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [314] Murphy, 2007, Targeting antioxidants to mitochondria by conjugation to lipophilic cations, Annu. Rev. Pharmacol. Toxicol. source

      Not checked in the free run.

    • cites [391] Smith, 1999, Selective targeting of an antioxidant to mitochondria, Eur. J. Biochem. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    MitoQ is an analog of ubiquinone and prevents lipid peroxidation in mitochondrial membranes.

    “whereas MitoQ is an analog of ubiquinone and prevents lipid peroxidation in mitochondrial membranes [314,315].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [314] Murphy, 2007, Targeting antioxidants to mitochondria by conjugation to lipophilic cations, Annu. Rev. Pharmacol. Toxicol. source

      Not checked in the free run.

    • cites [315] Kelso, 2001, Selective targeting of a redox-active ubiquinone to mitochondria within cells: Antioxidant and antiapoptotic properties, J. Biol. Chem. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    A number of studies have demonstrated extra- and intracellular ROS scavenging activities of vitamin C in different cell lines.

    “While a number of studies have demonstrated extra- and intracellular ROS scavenging activities of vitamin C in different cell lines [386,387]”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [386] Padh, 1991, Vitamin c: Newer insights into its biochemical functions, Nutr. Rev. source

      Not checked in the free run.

    • cites [387] Liang, 2001, Vitamin c transport systems of mammalian cells, Mol. Membr. Biol. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Electron leakage into the mitochondrial matrix is increased by stigmatellin/myxothiazol, resulting in increased generation of matrix mtROS.

    “Electron leakage into the mitochondrial matrix in consequence is increased, which results in increased generation of matrix mtROS [416].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [416] Stowe, 2009, Mitochondrial reactive oxygen species production in excitable cells: Modulators of mitochondrial and cell function, Antioxid. Redox Signal. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Blockade of rotenone to the binding site of ubiquinone leads to reduced electron flow to complex III and reverse electron flow through complex I into the mitochondrial matrix, leading to increased generation of matrix mtROS.

    “Blockade of rotenone to the binding site of ubiquinone leads to reduced electron flow to complex III and reverse electron flow through complex I into the mitochondrial matrix leading to increased ROS generation in this compartment, namely matrix mtROS [269,416,419,420,421].”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [269] Sena, 2013, Mitochondria are required for antigen-specific t cell activation through reactive oxygen species signaling, Immunity source

      Not checked in the free run.

    • cites [416] Stowe, 2009, Mitochondrial reactive oxygen species production in excitable cells: Modulators of mitochondrial and cell function, Antioxid. Redox Signal. source

      Not checked in the free run.

    • cites [419] Lambert, 2004, Inhibitors of the quinone-binding site allow rapid superoxide production from mitochondrial nadh:Ubiquinone oxidoreductase (complex i), J. Biol. Chem. source

      Not checked in the free run.

    • cites [420] Panov, 2005, Rotenone model of parkinson disease: Multiple brain mitochondria dysfunctions after short term systemic rotenone intoxication, J. Biol. Chem. source

      Not checked in the free run.

    • cites [421] Buckingham, 2002, Topology of superoxide production from different sites in the mitochondrial electron transport chain, J. Biol. Chem. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    While increasing matrix mtROS generation, rotenone effectively blocks ROS production by complex III and therefore generation of cytosolic mtROS.

    “Of note, while increasing matrix mtROS generation, the reduction of electron flow to complex III by rotenone effectively blocks ROS production by this complex and therefore generation of cytosolic mtROS [30,125].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [125] Pastukh, 2012, Perinuclear mitochondrial clustering creates an oxidant-rich nuclear domain required for hypoxia-induced transcription, Sci. Signal.

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S2 (checkable).

    Interpretationreasoning

    The highly complex and structured production of ROS in macrophages is often reduced to the phrase "ROS levels in macrophages".

    “In consequence, the highly complex and structured production of ROS in macrophages is often reduced to the phrase “ROS levels in macrophages”.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisereasoning

    Antimycin A acts as an inhibitor of the respiratory chain at complex III in terms of ATP production, but its influence on mtROS production is compartment-specific: it induces cytosolic mtROS production and abolishes matrix mtROS production.

    “Antimycin A acts as inhibitor of the respiratory chain at complex III in terms of ATP production, but its influence on mtROS production is compartment-specific. It induces cytosolic mtROS production instead of inhibiting it, while matrix mtROS production is abolished.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisereasoning

    Rotenone is a potent inducer of matrix mtROS generation and is widely used as a positive control for ROS probes targeted to the mitochondrial matrix, such as MitoSOX.

    “Therefore, rotenone is a potent inducer of the generation of matrix mtROS and hence is widely used as positive control for ROS probes targeted to the mitochondrial matrix, such as MitoSOX [30,270,271].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [270] Sazanov, 2014, The mechanism of coupling between electron transfer and proton translocation in respiratory complex i, J. Bioenerg. Biomembr. source

      Not checked in the free run.

    • cites [271] Orr, 2013, Inhibitors of ros production by the ubiquinone-binding site of mitochondrial complex i identified by chemical screening, Free Radic. Biol. Med. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In the IMS, leaked electrons react with O2 to generate ROS, which reach the cytosol.

    “In the IMS, leaked electrons react with O2 to generate ROS, which reach the cytosol [30,109,121,125,423].”
    Quote in the paperSubstantial claim0 of 5 sources checked
    • cites [30] Herb, M., Gluschko, A., Wiegmann, K., Farid, A., Wolf, A., Utermohlen, O., Krut, O., Kronke, M., and Schramm, M. (2019). Mitochondrial reactive oxygen species enable proinflammatory signaling through disulfide linkage of nemo. Sci. Signal., 12. source

      Not checked in the free run.

    • cites [109] Quinlan, 2012, Mitochondrial complex ii can generate reactive oxygen species at high rates in both the forward and reverse reactions, J. Biol. Chem. source

      Not checked in the free run.

    • cites [121] Han, 2003, Voltage-dependent anion channels control the release of the superoxide anion from mitochondria to cytosol, J. Biol. Chem. source

      Not checked in the free run.

    • cites [125] Pastukh, 2012, Perinuclear mitochondrial clustering creates an oxidant-rich nuclear domain required for hypoxia-induced transcription, Sci. Signal.

      Not checked in the free run.

    • cites [423] Chen, 2003, Production of reactive oxygen species by mitochondria: Central role of complex iii, J. Biol. Chem. source

      Not checked in the free run.

  • May combine two claims.

    Premisereasoning

    Myxothiazol and stigmatellin, like antimycin A, block electron flow to complex III, but at the Qo site, resulting in the opposite effect.

    “Myxothiazol and stigmatellin, like antimycin A, block electron flow to complex III. However, they do so at the Qo site of the complex, which results in the opposite effect of the one observed with antimycin A.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Antimycin A blocks the Qi site of complex III of the ETC, preventing electrons from entering the complex, leading to increased leakage of electrons into the IMS.

    “Antimycin A blocks the Qi site of complex III of the ETC [86,422], thereby preventing electrons from entering the complex, which leads to increased leakage of electrons into the IMS.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [86] Murphy, 2009, How mitochondria produce reactive oxygen species, Biochem. J. source

      Not checked in the free run.

    • cites [422] Bleier, 2013, Superoxide generation by complex iii: From mechanistic rationales to functional consequences, Biochim. Biophys. Acta source

      Not checked in the free run.

118 of 118

  • The paper relies on this without support.

    Only a few studies have performed in-depth ROS analyses and even fewer have identified the precise redox-regulated target molecules.

    “However, only a few studies have performed in-depth ROS analyses and even fewer have identified the precise redox-regulated target molecules.”

  • The paper relies on this without support.

    ROS are necessary for inflammasome activation.

    “It is also clear that ROS are necessary for inflammasome activation.”

  • The paper relies on this without support.

    The antimicrobial role for ROS against parasites in general is unquestioned.

    “While the antimicrobial role for ROS against parasites in general is unquestioned, the involved ROS sources remain controversial and need further investigations.”

  • The paper relies on this without support.

    The general role of ROS for inflammasome activation is unquestioned.

    “Like in nearly all fields of ROS-mediated immunity in infected macrophages, the general role of ROS for inflammasome activation is unquestioned.”

  • The paper relies on this without support.

    NAC is a commonly used global ROS scavenger that works extra- and intracellularly.

    “N-acetyl cysteine (NAC) is a commonly used global ROS scavenger, which works extra- and intracellularly.”

  • The paper relies on this without support.

    Vitamin C and its reduced form DHA are cell-impermeable and imported into the cell via Na+ cotransporters in the plasma membrane.

    “Vitamin C and its reduced form dehydroascorbic acid (DHA) are cell-impermeable and imported into the cell via Na+ cotransporters in the plasma membrane.”

  • The paper relies on this without support.

    Only a few studies have performed in-depth ROS analyses, and even fewer have provided mechanistic insights into redox-regulated targets.

    “However, only a few studies have performed in-depth ROS analyses, and even fewer have provided mechanistic insights into the redox-regulated targets.”
7 of 7

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • During an infective process, macrophages and other innate immunity cells especially produce ROS and RNS (•O2−, •NO and ONOO-), but excessive production can cause injury and toxicity to host cells.

    “During an infective process, macrophages and other innate immunity cells especially produce ROS and RNS (•O2−, •NO and ONOO—), but excessive production can cause injury and toxicity to host cells [121].”

    From The Role of Reactive Species on Innate Immunity (2022) · cites this paper as [121]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Reduced bacterial burden is called evidence of an antibacterial role for IRE1

    Major

    Both ROS-lowering treatment and IRE1 deficiency are said to reduce bacterial burden. Those reported directions conflict with the passage's conclusion that IRE1 has a ROS-mediated antibacterial role.

  • Parasite name differs from the cited reference

    Minor

    The text calls the organism "Toxoplasma cruzii," whereas the title of its cited reference names "trypanosoma cruzi."

  • Inflammasome receptor count conflicts with the list

    Minor

    The paper says five receptor proteins were identified, then names eight canonical inflammasome receptors: six NLR-family members, AIM2, and Pyrin.

  • NEMO cysteine residue differs from the cited reference

    Minor

    The text identifies the second redox-sensitive NEMO residue as Cys374, but the title of its cited reference identifies it as Cys347.

4 of 4

Cites

2 retracted

Flagged first, then by citations

Research works printed but not linked: 9

  1. Haber, 1997, The catalytic decomposition of hydrogen peroxide by iron salts, Proc. R. Soc. Lond. Ser. A Math. Phys. Sci., 147, 332
  2. Marinho, 2013, The cellular steady-state of h2o2: Latency concepts and gradients, Methods Enzym., 527, 3, 10.1016/B978-0-12-405882-8.00001-5 · DOI · Paper page
  3. Rhee, 2006, Cell signaling. H2o2, a necessary evil for cell signaling, Science, 312, 1882, 10.1126/science.1130481 · DOI · Paper page
  4. Lewis, 1990, Leukotrienes and other products of the 5-lipoxygenase pathway. Biochemistry and relation to pathobiology in human diseases, New Engl. J. Med., 323, 645, 10.1056/NEJM199009063231006 · DOI
  5. Omura, 1964, The carbon monoxide-binding pigment of liver microsomes. I. Evidence for its hemoprotein nature, J. Biol. Chem., 239, 2370, 10.1016/S0021-9258(20)82244-3 · DOI · Paper page
  6. Martinon, 2002, The inflammasome: A molecular platform triggering activation of inflammatory caspases and processing of proil-beta, Mol. Cell, 10, 417, 10.1016/S1097-2765(02)00599-3 · DOI · Paper page
  7. Cadenas, 2013, Chapter fifteen—Using hyper as a molecular probe to visualize hydrogen peroxide in living plant cells: A method with virtually unlimited potential in plant biology, Methods in Enzymology, Volume 527, 275, 10.1016/B978-0-12-405882-8.00015-5 · DOI
  8. Palmer, 1968, Studies on the respiratory chain-linked reduced nicotinamide adenine dinucleotide dehydrogenase. Xiv. Location of the sites of inhibition of rotenone, barbiturates, and piericidin by means of electron paramagnetic resonance spectroscopy, J. Biol. Chem., 243, 844, 10.1016/S0021-9258(19)81742-8 · DOI · Paper page
  9. Panov, 2005, Rotenone model of parkinson disease: Multiple brain mitochondria dysfunctions after short term systemic rotenone intoxication, J. Biol. Chem., 280, 42026, 10.1074/jbc.M508628200 · DOI · Paper page

Cited by

Sorted by citations

Authors
Herb, M., & Schramm, M.
Affiliations
University of Cologne (DE)
Corresponding author
Schramm, Michael
ORCID
Venue
Antioxidants
Publisher
MDPI AG
Published
2021-02-19
Volume
10
Issue
2
Pages
313
ISSN
2076-3921 (ISSN-L)
DOI
10.3390/antiox10020313
PMC
PMC7923022
OpenAlex
W3131190077
Cited by
633
References
417
Citations by year
2026: 85 · 2025: 173 · 2024: 129 · 2023: 129 · 2022: 87 · 2021: 30
FWCI
9.60
Type
journal article
Language
en
Open access
Licence
Locations
  • Antioxidants (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Kölner Universitäts PublikationsServer (Universität zu Köln) (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • Deutsche Forschungsgemeinschaft (1627/-21)
  • Köln Fortune (302/2020)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
  • Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
Keywords
reactive oxygen species, macrophages, antimicrobial immunity, redox regulation, inflammasome activation, immune signaling, ROS scavengers

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-06.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-06.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

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