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  1. AU="Massimiliano Stagi"
  2. AU="Graeme S. Cumming"
  3. AU="Field, Belinda G"
  4. AU="Erhart, Stephen"
  5. AU="Glenn Smith"
  6. AU="Shillingford, Shanelle R"
  7. AU="Ahn, Sung Soo"
  8. AU="Salih, Harith M"
  9. AU="Clayton, Philip A"
  10. AU="Soto, A. Garcia"
  11. AU="Jones, Daniel OB"
  12. AU="Chen, Maosheng"
  13. AU="Li, Zhengxi"
  14. AU="Toshiya Takahashi"
  15. AU=Hickey Chelsea L.
  16. AU="Badhrinarayanan, Shreya"
  17. AU="Milani, Liliana"
  18. AU="Reinhardt, Klaus"
  19. AU="Caudillo-Flores, Uriel"
  20. AU="Yin, Yizhen"
  21. AU=Kaushansky Kenneth
  22. AU="Golla, Jaya Prakash"
  23. AU="Penn, Marc S"
  24. AU="Montero, Vincent"
  25. AU="Etevenon, Pierre"
  26. AU="Hyseni, Agon"
  27. AU="Seitzman, Natalie"
  28. AU="Loukil, Abdelhalim"
  29. AU="Giammusso, Bruno"
  30. AU="Kaplan, Jonathan E"
  31. AU=Francolini Giulio
  32. AU="Yuhu Li"
  33. AU=Kim Moojung
  34. AU="Vise, Luciana M"
  35. AU="Marcinowska, Zuzanna"
  36. AU="Graff, Pablo"

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  1. Artikel ; Online: Impact of various buffers and weak bases on lysosomal and intracellular pH

    Jeffrey A. Kraut / Izaak J. Cheetham‐Wilkinson / Laura E. Swan / Massimiliano Stagi / Ira Kurtz

    FASEB BioAdvances, Vol 5, Iss 4, Pp 149-

    Implications for infectivity of SARS‐CoV‐2

    2023  Band 155

    Abstract: Abstract Acidification of the cellular lysosome is an important factor in infection of mammalian cells by SARS‐CoV‐2. Therefore, raising the pH of the lysosome would theoretically be beneficial in prevention or treatment of SARS‐CoV‐2 infection. Sodium ... ...

    Abstract Abstract Acidification of the cellular lysosome is an important factor in infection of mammalian cells by SARS‐CoV‐2. Therefore, raising the pH of the lysosome would theoretically be beneficial in prevention or treatment of SARS‐CoV‐2 infection. Sodium bicarbonate, carbicarb, and THAM are buffers that can be used clinically to provide base to patients. To examine whether these bases could raise lysosomal pH and therefore be a primary or adjunctive treatment of SARS‐CoV‐2 infection, we measured lysosomal and intracellular pH of mammalian cells after exposure to each of these bases. Mammalian HEK293 cells expressing RpH‐LAMP1‐3xFLAG, a ratiometric sensor of lysosomal luminal pH, were first exposed to Hepes which was then switched to sodium bicarbonate, carbicarb, or THAM and lysosomal pH measured. In bicarbonate buffer the mean lysosomal pH was 4.3 ± 0.1 (n = 20); p = NS versus Hepes (n = 20). The mean lysosomal pH in bicarbonate/carbonate was 4.3 ± 0.1 (n = 21) versus Hepes (n = 21), p = NS. In THAM buffer the mean lysosomal pH was 4.7 ± 0.07 (n = 20) versus Hepes (4.6 ± 0.1, n = 20), p = NS. In addition, there was no statistical difference between pHi in bicarbonate, carbicarb or THAM solutions. Using the membrane permeable base NH4Cl (5 mM), lysosomal pH increased significantly to 5.9 ± 0.1 (n = 21) compared to Hepes (4.5 ± 0.07, n = 21); p < 0.0001. Similarly, exposure to 1 mM hydroxychloroquine significantly increased the lysosomal pH to (5.9 ± 0.06, n = 20) versus Hepes (4.3 ± 0.1, n = 20), p < 0.0001. Separately steady‐state pHi was measured in HEK293 cells bathed in various buffers. In bicarbonate pHi was 7.29 ± 0.02 (n = 12) versus Hepes (7.45 ± 0.03, [n = 12]), p < 0.001. In cells bathed in carbicarb pHi was 7.27 ± 0.02 (n = 5) versus Hepes (7.43 ± 0.04, [n = 5]), p < 0.01. Cells bathed in THAM had a pHi of 7.25 ± 0.03 (n = 12) versus Hepes (7.44 ± 0.03 [n = 12]), p < 0.001. In addition, there was no statistical difference in pHi in bicarbonate, carbicarb or THAM solutions. The results of ...
    Schlagwörter alkalinization ; carbicarb ; lysosomal pH ; sars‐Covid‐ 19 ; sodium bicarbonate ; tham ; Biology (General) ; QH301-705.5
    Thema/Rubrik (Code) 572
    Sprache Englisch
    Erscheinungsdatum 2023-04-01T00:00:00Z
    Verlag Wiley
    Dokumenttyp Artikel ; Online
    Datenquelle BASE - Bielefeld Academic Search Engine (Lebenswissenschaftliche Auswahl)

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  2. Artikel ; Online: Effects of Human RelA Transgene on Murine Macrophage Inflammatory Responses

    Stamatia Papoutsopoulou / Lorna Morris / Andrew Bayliff / Thomas Mair / Hazel England / Massimiliano Stagi / François Bergey / Mohammad Tauqeer Alam / Raheleh Sheibani-Tezerji / Philip Rosenstiel / Werner Müller / Vitor A. P. Martins Dos Santos / Barry J. Campbell

    Biomedicines, Vol 10, Iss 757, p

    2022  Band 757

    Abstract: The NFκB transcription factors are major regulators of innate immune responses, and NFκB signal pathway dysregulation is linked to inflammatory disease. Here, we utilised bone marrow-derived macrophages from the p65-DsRedxp/IκBα-eGFP transgenic strain to ...

    Abstract The NFκB transcription factors are major regulators of innate immune responses, and NFκB signal pathway dysregulation is linked to inflammatory disease. Here, we utilised bone marrow-derived macrophages from the p65-DsRedxp/IκBα-eGFP transgenic strain to study the functional implication of xenogeneic (human) RelA(p65) protein introduced into the mouse genome. Confocal imaging showed that human RelA is expressed in the cells and can translocate to the nucleus following activation of Toll-like receptor 4. RNA sequencing of lipid A-stimulated macrophages, revealed that human RelA impacts on murine gene transcription, affecting both non-NFκB and NFκB target genes, including immediate-early and late response genes, e.g., Fos and Cxcl10 . Validation experiments on NFκB targets revealed markedly reduced mRNA levels, but similar kinetic profiles in transgenic cells compared to wild-type. Enrichment pathway analysis of differentially expressed genes revealed interferon and cytokine signaling were affected. These immune response pathways were also affected in macrophages treated with tumor necrosis factor. Data suggests that the presence of xenogeneic RelA protein likely has inhibitory activity, altering specific transcriptional profiles of key molecules involved in immune responses. It is therefore essential that this information be taken into consideration when designing and interpreting future experiments using this transgenic strain.
    Schlagwörter macrophage ; NFκB ; RelA(p65) ; inflammation ; lipid A ; lipopolysaccharide ; Biology (General) ; QH301-705.5
    Thema/Rubrik (Code) 616
    Sprache Englisch
    Erscheinungsdatum 2022-03-01T00:00:00Z
    Verlag MDPI AG
    Dokumenttyp Artikel ; Online
    Datenquelle BASE - Bielefeld Academic Search Engine (Lebenswissenschaftliche Auswahl)

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  3. Artikel ; Online: TREM2-transduced myeloid precursors mediate nervous tissue debris clearance and facilitate recovery in an animal model of multiple sclerosis.

    Kazuya Takahashi / Marco Prinz / Massimiliano Stagi / Olga Chechneva / Harald Neumann

    PLoS Medicine, Vol 4, Iss 4, p e

    2007  Band 124

    Abstract: BACKGROUND: In multiple sclerosis, inflammation can successfully be prevented, while promoting repair is still a major challenge. Microglial cells, the resident phagocytes of the central nervous system (CNS), are hematopoietic-derived myeloid cells and ... ...

    Abstract BACKGROUND: In multiple sclerosis, inflammation can successfully be prevented, while promoting repair is still a major challenge. Microglial cells, the resident phagocytes of the central nervous system (CNS), are hematopoietic-derived myeloid cells and express the triggering receptor expressed on myeloid cells 2 (TREM2), an innate immune receptor. Myeloid cells are an accessible source for ex vivo gene therapy. We investigated whether myeloid precursor cells genetically modified to express TREM2 affect the disease course of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. METHODS AND FINDINGS: EAE was induced in mice by immunization with a myelin autoantigen. Intravenous application of TREM2-transduced bone marrow-derived myeloid precursor cells at the EAE peak led to an amelioration of clinical symptoms, reduction in axonal damage, and prevention of further demyelination. TREM2-transduced myeloid cells applied intravenously migrated into the inflammatory spinal cord lesions of EAE-diseased mice, showed increased lysosomal and phagocytic activity, cleared degenerated myelin, and created an anti-inflammatory cytokine milieu within the CNS. CONCLUSIONS: Intravenously applied bone marrow-derived and TREM2-tranduced myeloid precursor cells limit tissue destruction and facilitate repair within the murine CNS by clearance of cellular debris during EAE. TREM2 is a new attractive target for promotion of repair and resolution of inflammation in multiple sclerosis and other neuroinflammatory diseases.
    Schlagwörter Medicine ; R
    Thema/Rubrik (Code) 616 ; 610
    Sprache Englisch
    Erscheinungsdatum 2007-04-01T00:00:00Z
    Verlag Public Library of Science (PLoS)
    Dokumenttyp Artikel ; Online
    Datenquelle BASE - Bielefeld Academic Search Engine (Lebenswissenschaftliche Auswahl)

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