2020

• Yamamoto, M;Miura, Y;Kyo, A;Hirayama, K;Kohno, T;Theisen-Kunde, D;Brinkmann, R and Honda, S: Selective retina therapy for subretinal fluid associated with choroidal nevus. Amer J Ophthalm Case Rep, no. 19, pp. 100794, 2020
  

  @article{yamamoto2020,
     author = {Yamamoto, M;Miura, Y;Kyo, A;Hirayama, K;Kohno, T;Theisen-Kunde, D;Brinkmann, R and Honda, S},
     title = {Selective retina therapy for subretinal fluid associated with choroidal nevus},
     journal = {Amer J Ophthalm Case Rep},
     volume = {19},
     pages = {100794},
     ISSN = {2451-9936},
  keywords = {Laser therapy, Choroidal tumor, Retinal pigment epithelium, Retinal disorder}
     DOI = {https://doi.org/10.1016/j.ajoc.2020.100794},
     
     year = {2020},
     type = {Journal Article}
  }
  

• Miura, Yoko: Retinal Pigment Epithelium Organ Culture. in Retinal Pigment Epithelium in Health and Disease, pp. 307-324, Springer, 2020
  

  @inbook{Miura2020,
     author = {Miura, Yoko},
     title = {Retinal Pigment Epithelium Organ Culture},
     booktitle = {Retinal Pigment Epithelium in Health and Disease},
     publisher = {Springer},
     pages = {307-324},
     year = {2020},
     type = {Book Section},
  URL = { https://link.springer.com/chapter/10.1007/978-3-030-28384-1_18}
  }
  

• Yamamoto, M;Miura, Y;Hirayama, K;;Kohno, T;Kabata, D;Theisen-Kunde, D;Brinkmann, R and Honda, S;: Predictive factors of outcome of selective retina therapy for diabetic macular edema. International Ophthalmology, 2020
  

  @article{Miura2020-2,
     author = {Yamamoto, M;Miura, Y;Hirayama, K;;Kohno, T;Kabata, D;Theisen-Kunde, D;Brinkmann, R and Honda, S;},
     title = {Predictive factors of outcome of selective retina therapy for diabetic macular edema},
     journal = {International Ophthalmology},
     ISSN = {1573-2630},
     
     url = {https://doi.org/10.1007/s10792-020-01288-6},
     year = {2020},
     type = {Journal Article}
  }
  

2019

• Pfäffle, C;Spahr, H;Kutzner, L;Burhan, S;Hilge, F;Miura, Y;Hüttmann, G and Hillmann, D: Simultaneous functional imaging of neuronal and photoreceptor layers in living human retina. Optics Letters 44(23), pp. 5671-5674, 2019
  

  @article{Pfäffle2019,
     author = {Pfäffle, C;Spahr, H;Kutzner, L;Burhan, S;Hilge, F;Miura, Y;Hüttmann, G and Hillmann, D},
     title = {Simultaneous functional imaging of neuronal and photoreceptor layers in living human retina},
     journal = {Optics Letters 44(23)},
   
     pages = {5671-5674},
     DOI = {10.1364/OL.44.005671},
     
     year = {2019},
     type = {Journal Article}
  }
  

• Miura, Y;Seifert, E;Rehra, J;Kern, K;Theisen-Kunde, D;Denton, M and Brinkmann, R: Real-time optoacoustic temperature determination on cell cultures during heat exposure: a feasibility study. Int J Hyperth, pp. 1-7, 2019
  

  @article{Miura2019/4,
     author = {Miura, Y;Seifert, E;Rehra, J;Kern, K;Theisen-Kunde, D;Denton, M and Brinkmann, R},
     title = {Real-time optoacoustic temperature determination on cell cultures during heat exposure: a feasibility study},
     journal = {Int J Hyperth},
     pages = {1-7},
     ISSN = {0265-6736},
    
     url = {https://doi.org/10.1080/02656736.2019.1590653},
     year = {2019},
     type = {Journal Article}
  }
  

• Klettner, A and Miura, Y: Porcine RPE/choroidal explant cultures. Weber B., Langmann T., Retinal Degeneration. Methods in Molecular Biology: 1834, pp. 109-118, 2019
  

  @book2019{Miura2019,
     author = {Klettner, A and  Miura, Y},
     title = {Porcine RPE/choroidal explant cultures},
     pages = {109-118 },
  journal = {Weber B., Langmann T., Retinal Degeneration. Methods in Molecular Biology: 1834},
     ISBN = {978-1-4939-8669-2},
    URL = {https://doi.org/10.1007/978-1-4939-8669-9_8},
     year = {2019},
     type = {Book}
  }
  

• Miura, Y; Draxinger, W; Grill, C; Pfeiffer, T; Grisanti, S and Huber, R: MHz-OCT for low latency virtual reality guided surgery: first wet lab experiments on ex-vivo porcine eye. in Proc. SPIE 11078, Optical Coherence Imaging Techniques and Imaging in Scattering Media III, 110780E, no. 11078, 2019
  

  @proceeding{Miura2019,
  author = { Miura, Y; Draxinger, W; Grill, C; Pfeiffer, T; Grisanti, S and Huber, R},
  title = { MHz-OCT for low latency virtual reality guided surgery: first wet lab experiments on ex-vivo porcine eye},
  volume = {11078},
  year = {2019},
  doi = {10.1117/12.2527123},
  URL = {https://doi.org/10.1117/12.2527123},
  keywords = {AG-Huber_OCT},
  booktitle =    {Proc. SPIE 11078, Optical Coherence Imaging Techniques and Imaging in Scattering Media III, 110780E},
  eprint = {}
  }
  

• Detrez, N;Miura, Y;Seifert, E;Theisen-Kunde, D and Brinkmann, R: Heating and optoacoustic temperature determination of cell cultures. in Proc. SPIE 11079, Medical Laser Applications and Laser-Tissue Interactions IX, no. 11079, SPIE, 2019
  

  @inproceedings{Detrez2019,
     author = {Detrez, N;Miura, Y;Seifert, E;Theisen-Kunde, D and Brinkmann, R},
     title = {Heating and optoacoustic temperature determination of cell cultures},
     publisher = {SPIE},
     volume = {11079},
     series = {European Conferences on Biomedical Optics},
  booktitle =    {Proc. SPIE 11079, Medical Laser Applications
  and Laser-Tissue Interactions IX},
     url = {https://doi.org/10.1117/12.2527024},
  keywords = {Laser, Noninvasive thermometry, hyperthermia, temperature measurement, photoacoustics}, optoacoustics,
     year = {2019},
     type = {Conference Proceeding}
  }
  

• Hirayama, K;Manabu Yamamoto, M; Takeya Kohno, T; Miura, Y; Brinkmann, R; Shiraki,K;Theisen-Kunde, D; and Honda, S;: hange in the Thickness of Retinal Layers after Selective Retina Therapy (SRT) in Patients with Central Serous Chorioretinopathy. Osaka City Med., no. 65, pp. 55-63, 2019
  

  @article{Miura2019,
     author = {Hirayama, K;Manabu Yamamoto, M; Takeya Kohno, T; Miura, Y; Brinkmann, R;  Shiraki,K;Theisen-Kunde, D; and Honda, S;},
     title = {Change in the Thickness of Retinal Layers after Selective Retina
  Therapy (SRT) in Patients with Central Serous Chorioretinopathy},
     journal = {Osaka City Med.},
     volume = {65},
     pages = {55-63},
     url = {http://dlisv03.media.osaka-cu.ac.jp/il/meta_pub/G0000438repository_00306096-65-1-55},
     year = {2019},
     type = {Journal Article}
  }
  

• Miura, Y;Bernstein, P S;Dysli, C;Sauer, L and Zinkernagel, M: Fluorophores in the Eye. in Fluorescence Lifetime Imaging Ophthalmoscopy, pp. 35-48, Springer International Publishing, Cham, 2019
  

  @inbook{Miura2019,
     author = {Miura, Y;Bernstein, P S;Dysli, C;Sauer, L and Zinkernagel, M},
     title = {Fluorophores in the Eye},
     booktitle = {Fluorescence Lifetime Imaging Ophthalmoscopy},
     editor = {Zinkernagel, Martin and Dysli, Chantal},
     publisher = {Springer International Publishing},
     address = {Cham},
     pages = {35-48},
     ISBN = {978-3-030-22878-1},
  Keywords = {Retinoid cycle Lipofuscin Macular pigment Collagen/elastin Redox coenzyme Melanin Lipid peroxidation endproducts Advanced glycation endproducts (AGE) },
    
     url = {https://doi.org/10.1007/978-3-030-22878-1_7},
     year = {2019},
     type = {Book Section}
  }
  

• Hutfilz, A;Sonntag, S;Lewke, B;Theisen-Kunde, D;Grisanti, S;Brinkmann, R and Miura, Y: Fluorescence Lifetime Imaging Ophthalmoscopy of the Retinal Pigment Epithelium During Wound Healing After Laser Irradiation. Translational Vision Science & Technology, no. 8(5), 2019
  

  @article{Hutfilz2019,
     author = {Hutfilz, A;Sonntag, S;Lewke, B;Theisen-Kunde, D;Grisanti, S;Brinkmann, R and Miura, Y},
     title = {Fluorescence Lifetime Imaging Ophthalmoscopy of the Retinal Pigment Epithelium During Wound Healing After Laser Irradiation},
     journal = {Translational Vision Science & Technology},
     volume = {8(5)},
   
     ISSN = {2164-2591},
     DOI = {10.1167/tvst.8.5.12},
     year = {2019},
     type = {Journal Article}
  }
  

• Miura, Y;Lewke, B;Hutfilz, A and Brinkmann, R: Change in fluorescence lifetime of retinal pigment epithelium under oxidative stress. Nippon Ganka Gakkai Zasshi 123 (2), pp. 105-114, 2019
  

  @article{Miura2019/3,
     
     author = {Miura, Y;Lewke, B;Hutfilz, A and Brinkmann, R},
     title = {Change in fluorescence lifetime of retinal pigment epithelium under oxidative stress},
     journal = {Nippon Ganka Gakkai Zasshi } {123 (2)},
    
     pages = {105-114},
     url = {http://journal.nichigan.or.jp/Disp?style=abst&vol=123&year=2019&mag=0&number=2&start=105},
     year = {2019},
     type = {Journal Article}
  }
  
  

• Rudolf, M; Curcio, C A; Schlözer-Schrehardt, U; Sefat, A M M; Tura, A; Aherrahrou, Z; Brinkmann, M; Grisanti, S; Miura, Y and Ranjbar, M: Apolipoprotein A-I mimetic peptide L-4F removes Bruch`s membrane lipids in aged nonhuman primates. Invest Ophthalmol Vis Sci, pp. 461-472, 2019
  

  @article{Miura2019-2,
     author = {Rudolf, M; Curcio, C A; Schlözer-Schrehardt, U; Sefat, A M M; Tura, A; Aherrahrou, Z; Brinkmann, M; Grisanti, S;  Miura, Y and Ranjbar, M},
     title = {Apolipoprotein A-I mimetic peptide L-4F removes Bruch's membrane lipids in aged nonhuman primates},
     journal = {Invest Ophthalmol Vis Sci},
     pages = {461-472},
    
     url = {https://www.ncbi.nlm.nih.gov/pubmed/30707219},
     year = {2019},
     type = {Journal Article}
  }
  

• Draxinger, D; Miura, Y; Grill, C; Pfeiffer, T and Huber, R: A real-time video-rate 4D MHz-OCT microscope with high definition and low latency virtual reality display. in Proc. SPIE 11078, Optical Coherence Imaging Techniques and Imaging in Scattering Media III, 1107802, no. 11078, 2019
  

  @proceeding{Draxinger2019,
  author = {Draxinger, D; Miura, Y; Grill, C; Pfeiffer, T and Huber, R},
  title = {A real-time video-rate 4D MHz-OCT microscope with high definition and low latency virtual reality display},
  volume = {11078},
  year = {2019},
  
  URL = {https://doi.org/10.1117/12.2527177},
  keywords = {AG-Huber_OCT},
  booktitle =    {Proc. SPIE 11078, Optical Coherence Imaging Techniques and Imaging in Scattering Media III, 1107802},
  eprint = {}
  }
  

2018

• Miura, Y: Two-Photon Microscopy (TPM) and Fluorescence Lifetime Imaging Microscopy (FLIM) of Retinal Pigment Epithelium (RPE) of Mice In Vivo. in Mouse Retinal Phenotyping: Methods and Protocols, pp. 73-88, Springer New York, New York, NY, 2018
  

  @inbook{Miura2018,
     author = {Miura, Y},
     title = {Two-Photon Microscopy (TPM) and Fluorescence Lifetime Imaging Microscopy (FLIM) of Retinal Pigment Epithelium (RPE) of Mice In Vivo},
     booktitle = {Mouse Retinal Phenotyping: Methods and Protocols},
     editor = {Tanimoto, Naoyuki},
     publisher = {Springer New York},
    
     pages = {73-88},
     ISBN = {978-1-4939-7720-8},
     url={https://doi.org/10.1007/978-1-4939-7720-8_5},
     year = {2018},
     type = {Book Section}
  }
  

• Seifert, E; Tode, J; Pielen, A; Theisen-Kunde, D; Framme, C; Roider, J; Miura, Y; Birngruber, R and Brinkmann, R: Selective retina therapy: toward an optically controlled automatic dosing. J Biomed Opt 23(11), pp. 1-12, 2018
  

  @article{seifert2018,
     author = {Seifert, E; Tode, J; Pielen, A; Theisen-Kunde, D; Framme, C; Roider, J; Miura, Y; Birngruber, R and Brinkmann, R},
     title = {Selective retina therapy: toward an optically controlled automatic dosing},
     journal = {J Biomed Opt} {23(11)},
     
     pages = {1-12},
     ISSN = {1560-2281 (Electronic)
  1083-3668 (Linking)},
     DOI = {10.1117/1.JBO.23.11.115002},   
  keywords = {algorithm, lasers in medicine, ophthalmology, retinal pigment epithelium, selective retina therapy, selectivity}
     year = {2018},
     type = {Journal Article}
  }
  

• Kern, K; Mertineit, C L; Brinkmann, R and Miura, Y: Expression of heat shock protein 70 and cell death kinetics after different thermal impacts on cultured retinal pigment epithelial cells. Exp Eye Res 170, pp. 117-126, 2018
  

  @article{Miura2018,
     author = {Kern, K; Mertineit, C L; Brinkmann, R and Miura, Y},
     title = {Expression of heat shock protein 70 and cell death kinetics after different thermal impacts on cultured retinal pigment epithelial cells},
     journal = {Exp Eye Res} {170},
    
     pages = {117-126},
     ISSN = {1096-0007 (Electronic)
  0014-4835 (Linking)},
     DOI = {10.1016/j.exer.2018.02.013},
     year = {2018},
     type = {Journal Article}
  }
  

2017

• Baade, A; von der Burchard, C; Lawin, M; Koinzer, S; Schmarbeck, B; Schlott, K; Miura, Y; Roider, J; Birngruber, R and Brinkmann, R: Power-controlled temperature guided retinal laser therapy. J Biomed Opt 22(11), pp. 1-11, 2017
  

  @article{Baade2017,
     author = {Baade, A; von der Burchard, C; Lawin, M; Koinzer, S; Schmarbeck, B; Schlott, K; Miura, Y; Roider, J; Birngruber, R and Brinkmann, R},
     title = {Power-controlled temperature guided retinal laser therapy},
     journal = {J Biomed Opt} {22(11)},
     
     pages = {1-11},
     ISSN = {1083-3668},
     DOI = {10.1117/1.jbo.22.11.118001},
     year = {2017},
     type = {Journal Article}
  }
  

• Miura, Y; Pruessner, J; Mertineit, C L; Kern, K; Muenter, M; Moltmann, M; Danicke, V and Brinkmann, R: Continuous-wave Thulium Laser for Heating Cultured Cells to Investigate Cellular Thermal Effects. J Vis Exp 30(124), 2017
  

  @article{Miura2017,
     author = {Miura, Y; Pruessner, J; Mertineit, C L; Kern, K; Muenter, M; Moltmann, M; Danicke, V and Brinkmann, R},
     title = {Continuous-wave Thulium Laser for Heating Cultured Cells to Investigate Cellular Thermal Effects},
     journal = {J Vis Exp} {30(124)},
     
     ISSN = {1940-087x},
     DOI = {10.3791/54326},
     year = {2017},
     type = {Journal Article},
  
  

• Rudolf, M; Mir Mohi Sefat, A; Miura, Y; Tura, A; Raasch, W; Ranjbar, M; Grisanti, S; Aherrahrou, Z; Wagner, A; Messinger, J D; Garber, D W; Anantharamaiah, G M and Curcio, C A: ApoA-I Mimetic Peptide 4F Reduces Age-Related Lipid Deposition in Murine Bruch`s Membrane and Causes Its Structural Remodeling. Curr Eye Res 3, pp. 1-12, 2017
  

  @article{Miura2017,
     author = {Rudolf, M; Mir Mohi Sefat, A; Miura, Y; Tura, A; Raasch, W; Ranjbar, M; Grisanti, S; Aherrahrou, Z; Wagner, A; Messinger, J D; Garber, D W; Anantharamaiah, G M and Curcio, C A},
     title = {ApoA-I Mimetic Peptide 4F Reduces Age-Related Lipid Deposition in Murine Bruch's Membrane and Causes Its Structural Remodeling},
     journal = {Curr Eye Res} {3},
     pages = {1-12},
     ISSN = {0271-3683},
     DOI = {10.1080/02713683.2017.1370118},
     year = {2017},
     type = {Journal Article},
  
  }
  

2016

• Yasui, Ayako and Yamamoto, Manabu and Hirayama, Kumiko and Shiraki, Kunihiko and Theisen-Kunde, Dirk and Brinkmann, Ralf and Miura, Yoko and Kohno, Takeya: Retinal sensitivity after selective retina therapy (SRT) on patients with central serous chorioretinopathy. Graefe`s Archive for Clinical and Experimental Ophthalmology, pp. 1-12, 2016
  

  @article{Yasui2016,
     author = {Yasui, Ayako and Yamamoto, Manabu and Hirayama, Kumiko and Shiraki, Kunihiko and Theisen-Kunde, Dirk and Brinkmann, Ralf and Miura, Yoko and Kohno, Takeya},
     title = {Retinal sensitivity after selective retina therapy (SRT) on patients with central serous chorioretinopathy},
     journal = {Graefe's Archive for Clinical and Experimental Ophthalmology},
     pages = {1-12},
     abstract = {To assess retinal sensitivity after selective retina therapy (SRT) in patients with central serous chorioretinopathy (CSCR).},
     ISSN = {1435-702X},
     url = {http://dx.doi.org/10.1007/s00417-016-3441-8},
     year = {2016},
     type = {Journal Article}
  }
  

• Ranjbar, Mahdy and Brinkmann, Max Philipp and Tura, Aysegül and Rudolf, Martin and Miura, Yoko and Grisanti, Salvatore: Ranibizumab interacts with the VEGF-A/VEGFR-2 signaling pathway in human RPE cells at different levels. Cytokine, no. 83, pp. 210-216, 2016
  

  @article{Ranjbar2016,
     author = {Ranjbar, Mahdy and Brinkmann, Max Philipp and Tura, Aysegül and Rudolf, Martin and Miura, Yoko and Grisanti, Salvatore},
     title = {Ranibizumab interacts with the VEGF-A/VEGFR-2 signaling pathway in human RPE cells at different levels},
     journal = {Cytokine},
     volume = {83},
     pages = {210-216},
     abstract = {Vascular endothelial growth factor (VEGF) secreted by the retinal pigment epithelium (RPE) plays an important role in ocular homeostasis, but also in diseases, most notably age-related macular degeneration (AMD). To date, anti-VEGF drugs like ranibizumab have been shown to be most effective in treating these pathologic conditions. However, clinical trials suggest that the RPE could degenerate and perish through anti-VEGF treatment. Herein, we evaluated possible pathways and outcomes of the interaction between ranibizumab and human RPE cells (ARPE-19). Results indicate that ranibizumab affects the VEGF-A metabolism in RPE cells from an extra- as well as intracellular site. The drug is taken up into the cells, with the VEGF receptor 2 (VEGFR-2) being involved, and decreases VEGF-A protein levels within the cells as well as extracellularly. Oxidative stress plays a key role in various inflammatory disorders of the eye. Our results suggest that oxidative stress inhibits RPE cell proliferation. This anti-proliferative effect on RPE cells is significantly enhanced through ranibizumab, which does not inhibit RPE cell proliferation substantially in absence of relevant oxidative stress. Therefore, we emphasize that anti-VEGF treatment should be selected carefully in AMD patients with preexistent extensive RPE atrophy.},
     keywords = {Ranibizumab
  RPE
  VEGF-A
  VEGFR-2
  Oxidative stress},
     ISSN = {1043-4666},
     url = {http://www.sciencedirect.com/science/article/pii/S1043466616300722},
     year = {2016},
     type = {Journal Article}
  }
  

• Ranjbar, M. and Brinkmann, M. P. and Zapf, D. and Miura, Y. and Rudolf, M. and Grisanti, S.: Fc Receptor Inhibition Reduces Susceptibility to Oxidative Stress in Human RPE Cells Treated with Bevacizumab, but not Aflibercept. Cell Physiol Biochem, no. 38, pp. 737-47, 2016
  

  @article{Ranjbar2016,
     author = {Ranjbar, M. and Brinkmann, M. P. and Zapf, D. and Miura, Y. and Rudolf, M. and Grisanti, S.},
     title = {Fc Receptor Inhibition Reduces Susceptibility to Oxidative Stress in Human RPE Cells Treated with Bevacizumab, but not Aflibercept},
     journal = {Cell Physiol Biochem},
     volume = {38},
     number = {2},
     pages = {737-47},
     note = {1421-9778
  Ranjbar, Mahdy
  Brinkmann, Max Philipp
  Zapf, Dorinja
  Miura, Yoko
  Rudolf, Martin
  Grisanti, Salvatore
  Journal Article
  Switzerland
  Cell Physiol Biochem. 2016;38(2):737-47. doi: 10.1159/000443030. Epub 2016 Feb 15.},
     abstract = {BACKGROUND/AIMS: VEGF-A is induced by oxidative stress, and functions as a survival factor for various cell types, including retinal pigment epithelial (RPE) cells. Anti-vascular endothelial growth factor (VEGF) drugs like aflibercept and bevacizumab have shown to be most effective in treating neovascular age-related macular degeneration (AMD), however uptake of the drugs might lead to interference with cell physiology. Herein, we evaluated the significance of the Fc receptor (FcR) within this context and moreover explored the impact of VEGF inhibition under normal conditions as well as under oxidative stress, in terms of potential adverse effects. METHODS: ARPE-19 (human RPE) cells were treated with aflibercept and bevacizumab in presence or absence of H2O2 as oxidative stress stimulus. After 24h cells were evaluated for drug uptake, VEGF-A expression and secretion, levels of intracellular reactive oxygen species (ROS) as well as cell proliferation. Experiments were repeated with cells being pre-incubated with an FcR inhibitor prior to drug application. RESULTS: Both drugs inhibited extracellular levels of VEGF-A and were taken up into the RPE, resulting in significantly reduced intracellular levels of VEGF-A. When oxidative stress was applied, intracellular ROS levels in cells treated with both drugs rose, and cell proliferation was reduced. Prior incubation with the FcR inhibitor lessened the uptake of bevacizumab, but not aflibercept into RPE cells, and simultaneously enhanced cell survival under oxidative stress conditions. CONCLUSIONS: Our results indicate that uptake and accumulation of aflibercept and bevacizumab within RPE cells affect the intracellular VEGF-A metabolism negatively, leading to a biologically relevant reduced cell survival under oxidative stress. The FcR plays a substantial role in the uptake of bevacizumab, but not aflibercept, which allows an enhanced RPE cell survival through FcR blockage in an environment dominated by oxidative stress, as clinically significant for various inflammatory retinal disorders.},
     ISSN = {1015-8987},
     DOI = {10.1159/000443030},
     year = {2016},
     type = {Journal Article}
  }
  

2014

• Iwami, H. and Pruessner, J. and Shiraki, K. and Brinkmann, R. and Miura, Y.: Protective effect of a laser-induced sub-lethal temperature rise on RPE cells from oxidative stress. Exp Eye Res, no. 124c, pp. 37-47, 2014
  

  @article{Iwami2014,
     author = {Iwami, H. and Pruessner, J. and Shiraki, K. and Brinkmann, R. and Miura, Y.},
     title = {Protective effect of a laser-induced sub-lethal temperature rise on RPE cells from oxidative stress},
     journal = {Exp Eye Res},
     volume = {124c},
     pages = {37-47},
     note = {1096-0007
  Iwami, Hisashi
  Pruessner, Joachim
  Shiraki, Kunihiko
  Brinkmann, Ralf
  Miura, Yoko
  Journal article
  Exp Eye Res. 2014 May 5;124C:37-47. doi: 10.1016/j.exer.2014.04.014.},
     abstract = {Recently introduced new technologies that enable temperature-controlled laser irradiation on the RPE allowed us to investigate temperature-resolved RPE cell responses. In this study we aimed primarily to establish an experimental setup that can realize laser irradiation on RPE cell culture with the similar temperature distribution as in the clinical application, with a precise time/temperature history. With this setup, we conducted investigations to elucidate the temperature-dependent RPE cell biochemical responses and the effect of transient hyperthermia on the responses of RPE cells to the secondary-exposed oxidative stress. Porcine RPE cells cultivated in a culture dish (inner diameter = 30 mm) with culture medium were used, on which laser radiation (lambda = 1940 nm, spot diameter = 30 mm) over 10 s was applied as a heat source. The irradiation provides a radially decreasing temperature profile which is close to a Gaussian shape with the highest temperature in the center. Power setting for irradiation was determined such that the peak temperature (Tmax) in the center of the laser spot at the cells reaches from 40 degrees C to 58 degrees C (40, 43, 46, 50, 58 degrees C). Cell viability was investigated with ethidium homodimer III staining at the time points of 3 and 24 h following laser irradiation. Twenty four hours after laser irradiation the cells were exposed to hydrogen peroxide (H2O2) for 5 h, followed by the measurement of intracellular glutathione, intracellular 4-hydroxynonenal (HNE) protein adducts, and secreted vascular endothelial growth factor (VEGF). The mean temperature threshold for RPE cell death after 3 h was found to be around 52 degrees C, and for 24 h around 50 degrees C with the current irradiation setting. A sub-lethal preconditioning on Tmax = 43 degrees C significantly induced the reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio, and decreased H2O2-induced increase of intracellular 4-HNE protein adducts. Although sub-lethal hyperthermia (Tmax = 40 degrees C, 43 degrees C, and 46 degrees C) caused a slight increase of VEGF secretion in 6 h directly following irradiation, secondary exposed H2O2-induced VEGF secretion was significantly reduced in the sub-lethally preheated groups, where the largest effect was seen following the irradiation with Tmax = 43 degrees C. In summary, the current results suggest that sub-lethal thermal laser irradiation on the RPE at Tmax = 43 degrees C for 10 s enhances cell defense system against oxidative stress, with increasing the GSH/GSSG ratio. Together with the results that the decreased amount of H2O2-induced 4-HNE in sub-lethally preheated RPE cells was accompanied by the lower secretion of VEGF, it is also strongly suggested that the sub-lethal hyperthermia may modify RPE cell functionality to protect RPE cells from oxidative stress and associated functional decrease, which are considered to play a significant role in the pathogenesis of age-related macular degeneration and other chorioretinal degenerative diseases.},
     ISSN = {0014-4835},
     DOI = {10.1016/j.exer.2014.04.014},
     year = {2014},
     type = {Journal Article}
  }
  

• Rudolf, M. and Mohi, A. and Dettbarn, M. C. and Miura, Y. and Aherrahrou, Z. and Ranjbar, M. and Mutus, B. and Knobloch, J. K.: Detection of esterified cholesterol in murine Bruch`s membrane wholemounts with a perfringolysin O-based cholesterol marker. Invest Ophthalmol Vis Sci, no. 55, pp. 4759-67, 2014
  

  @article{Rudolf2014,
     author = {Rudolf, M. and Mohi, A. and Dettbarn, M. C. and Miura, Y. and Aherrahrou, Z. and Ranjbar, M. and Mutus, B. and Knobloch, J. K.},
     title = {Detection of esterified cholesterol in murine Bruch's membrane wholemounts with a perfringolysin O-based cholesterol marker},
     journal = {Invest Ophthalmol Vis Sci},
     volume = {55},
     number = {8},
     pages = {4759-67},
     note = {1552-5783
  Rudolf, Martin
  Mohi, Armin
  Dettbarn, Marie C
  Miura, Yoko
  Aherrahrou, Zouhair
  Ranjbar, Mahdy
  Mutus, Bulent
  Knobloch, Johannes K M
  Journal Article
  Research Support, Non-U.S. Gov't
  United States
  Invest Ophthalmol Vis Sci. 2014 Jul 1;55(8):4759-67. doi: 10.1167/iovs.14-14311.},
     abstract = {PURPOSE: To investigate the effects of Bruch's membrane (BrM) neutral lipid deposition in mouse models and its significance to aging and age-related macular degeneration, it is essential to reliably detect small quantities of neutral lipids including esterified cholesterol (EC). In chorioretinal sections and BrM wholemounts, we tested a novel fluorescent cholesterol marker based on the bacterial toxin perfringolysin O (PFO) and compared results with those obtained with the classic cholesterol dye filipin. METHODS: An engineered plasmid containing the specific cholesterol binding domain (D4) of PFO fused to green fluorescent protein (GFP) was expressed in cultured E. coli, isolated, purified, and concentrated. A total of 150 BrM-choroid wholemounts and chorioretinal sections of 11- to 13-month-old ApoE(null) mice were prepared and stained with PFO/D4-GFP or filipin for EC. Samples were examined by epifluorescence microscopy. RESULTS: The fluorescence intensity of PFO/D4-GFP was strong, stable, and, if small quantities of EC were present, superior to filipin. In all specimens, we could sharply locate the PFO/D4-GFP signal to BrM. A semiquantitative evaluation of BrM lipid deposition is possible by measuring PFO/D4-GFP fluorescence intensity. CONCLUSIONS: The use of PFO/D4-GFP allowed a robust and direct detection of EC in aged murine BrM. In wholemount samples, its strong and stable fluorescence facilitated a semiquantitative evaluation of BrM-EC content over a large area. The patterns of EC deposition in murine BrM wholemounts are comparable with findings in human BrM wholemounts. Perfringolysin O/D4-GFP could be an important tool for investigating the effects of BrM lipid deposition in mouse models.},
     keywords = {Aging/*metabolism/pathology
  Animals
  Bacterial Toxins
  Bruch Membrane/*metabolism/ultrastructure
  Cells, Cultured
  Cholesterol Esters/*metabolism
  Clostridium perfringens
  Disease Models, Animal
  Feasibility Studies
  Female
  Hemolysin Proteins/*diagnostic use
  Humans
  Macular Degeneration/*diagnosis/metabolism
  Mice
  Mice, Inbred C57BL
  Microscopy, Electron, Transmission},
     ISSN = {0146-0404},
     DOI = {10.1167/iovs.14-14311},
     year = {2014},
     type = {Journal Article}
  }
  

• Koinzer, S. and Caliebe, A. and Portz, L. and Saeger, M. and Miura, Y. and Schlott, K. and Brinkmann, R. and Roider, J.: Comprehensive detection, grading, and growth behavior evaluation of subthreshold and low intensity photocoagulation lesions by optical coherence tomographic and infrared image analysis. Biomed Res Int, no. 2014, pp. 492679, 2014
  

  @article{Koinzer2014,
     author = {Koinzer, S. and Caliebe, A. and Portz, L. and Saeger, M. and Miura, Y. and Schlott, K. and Brinkmann, R. and Roider, J.},
     title = {Comprehensive detection, grading, and growth behavior evaluation of subthreshold and low intensity photocoagulation lesions by optical coherence tomographic and infrared image analysis},
     journal = {Biomed Res Int},
     volume = {2014},
     pages = {492679},
     note = {2314-6141
  Koinzer, Stefan
  Caliebe, Amke
  Portz, Lea
  Saeger, Mark
  Miura, Yoko
  Schlott, Kerstin
  Brinkmann, Ralf
  Roider, Johann
  Journal Article
  Research Support, Non-U.S. Gov't
  United States
  Biomed Res Int. 2014;2014:492679. doi: 10.1155/2014/492679. Epub 2014 May 12.},
     abstract = {PURPOSE: To correlate the long-term clinical effect of photocoagulation lesions after 6 months, as measured by their retinal damage size, to exposure parameters. We used optical coherence tomographic (OCT)-based lesion classes in order to detect and assess clinically invisible and mild lesions. METHODS: In this prospective study, 488 photocoagulation lesions were imaged in 20 patients. We varied irradiation diameters (100/300 microm), exposure-times (20-200 ms), and power. Intensities were classified in OCT images after one hour, and we evaluated OCT and infrared (IR) images over six months after exposure. RESULTS: For six consecutive OCT-based lesion classes, the following parameters increased with the class: ophthalmoscopic, OCT and IR visibility rate, fundus and OCT diameter, and IR area, but not irradiation power. OCT diameters correlated with exposure-time, irradiation diameter, and OCT class. OCT classes discriminated the largest bandwidth of OCT diameters. CONCLUSION: OCT classes represent objective and valid endpoints of photocoagulation intensity even for "subthreshold" intensities. They are suitable to calculate the treated retinal area. As the area is critical for treatment efficacy, OCT classes are useful to define treatment intensity, calculate necessary lesion numbers, and universally categorize lesions in clinical studies.},
     DOI = {10.1155/2014/492679},
     url = {http://dx.doi.org/10.1155/2014/492679},
     year = {2014},
     type = {Journal Article}
  }
  

2013

• Miura, Y. and Huettmann, G. and Orzekowsky-Schroeder, R. and Steven, P. and Szaszak, M. and Koop, N. and Brinkmann, R.: Two-Photon Microscopy and Fluorescence Lifetime Imaging of Retinal Pigment Epithelial Cells under Oxidative Stress. Invest Ophthalmol Vis Sci, 2013
  

  @article{Miura2013,
     author = {Miura, Y. and Huettmann, G. and Orzekowsky-Schroeder, R. and Steven, P. and Szaszak, M. and Koop, N. and Brinkmann, R.},
     title = {Two-Photon Microscopy and Fluorescence Lifetime Imaging of Retinal Pigment Epithelial Cells under Oxidative Stress},
     journal = {Invest Ophthalmol Vis Sci},
     note = {Miura, Yoko
  Huettmann, Gereon
  Orzekowsky-Schroeder, Regina
  Steven, Philipp
  Szaszak, Marta
  Koop, Norbert
  Brinkmann, Ralf
  ENG
  2013/04/06 06:00
  Invest Ophthalmol Vis Sci. 2013 Apr 4. pii: iovs.13-11808v1. doi: 10.1167/iovs.13-11808.},
     abstract = {PURPOSE: The aim of this study was to investigate the autofluorescence (AF) of the RPE with two-photon microscopy (TPM) and fluorescence lifetime imaging (FLIM) under normal and oxidative stress conditions. METHODS: Porcine RPE-choroid explants were used for investigation. The RPE-choroid tissue was preserved in a perfusion organ culture system. Oxidative stress was induced by laser photocoagulation with frequency-doubled Nd:YAG laser (532 nm) and by exposure to different concentrations (0, 1, 10 mM) of ferrous sulfate (FeSO4) for 1 hr. At indicated time points after exposure, the tissue was examined with TPM and FLIM. Intracellular reactive oxygen species around the photocoagulation lesion were detected with chloromethyl-2'7'-dichlorofluorescein diacetate (CM-H2DCFDA). Melanosomes were isolated from RPE cells and its fluorescence properties were investigated under normal and oxidized conditions. RESULTS: Under normal condition, AF in RPE cells with TPM is mostly originated from melanosomes, which has a very short fluorescence lifetime (FLT) (mean=117 ps). Under oxidative stress induced by laser irradiation and FeSO4 exposure, bright granular AF appears inside and around RPE cells, whose FLT is significantly longer (mean=1388 ps) than the FLT of the melanosome-AF. Excitation and emission peaks are found at 710-750 nm and 450-500 nm, respectively. Oxidative stress increases the fluorescence intensity of the melanosomes but does not change their FLT. CONCLUSION: TPM reveals acute oxidative stress-induced bright AF granules inside and around RPE cells which can be clearly discriminated from melanosomes by FLIM. TPM combined with FLIM is a useful tool of live-cell analysis to investigate functional alterations of the RPE.},
     year = {2013}
  }
  

• Miura, Y and Huettmann, G and Orzekowsky-Schroeder, R and Steven, P and Szaszák, M and Koop, N and Brinkmann, R: Two-photon Microscopy and Fluorescence Lifetime Analysis of Lipid Peroxidation Product in Photoreceptor Outer Segment and in Retinal Pigment Epithelial Cell. ARVO Meeting Abstracts, March 26, 2012, 2013
  

  @misc{Miura2013,
     author = {Miura, Y and Huettmann, G and Orzekowsky-Schroeder, R and Steven, P and Szaszák, M and Koop, N and Brinkmann, R },
     title = {Two-photon Microscopy and Fluorescence Lifetime Analysis of Lipid Peroxidation Product in Photoreceptor Outer Segment and in Retinal Pigment Epithelial Cell},
     publisher = {ARVO Meeting Abstracts},
     month = {March 26, 2012 },
     url = {http://www.abstractsonline.com/Plan/ViewAbstract.aspx?sKey=57630548-893d-4e45-9ddc-b6f547dd4ff0&cKey=d08a30bc-fe98-40a2-8a1c-1b171e4becd3&mKey=f0fce029-9bf8-4e7c-b48e-9ff7711d4a0e},
     year = {2013},
     type = {Poster}
  }
  

• Brinkmann, Ralf and Iwami, Hisashi and Pruessner, Joachim and Danicke, Veit and Miura, Yoko: Temperature-dependent response of retinal pigment epithelial cells to laser irradiation. Invest. Ophthalmol. Vis. Sci., no. 54, pp. 1809-, 2013
  

  @article{Brinkmann2013,
     author = {Brinkmann, Ralf and Iwami, Hisashi and Pruessner, Joachim and Danicke, Veit and Miura, Yoko},
     title = {Temperature-dependent response of retinal pigment epithelial cells to laser irradiation},
     journal = {Invest. Ophthalmol. Vis. Sci.},
     volume = {54},
     number = {6},
     pages = {1809-},
     abstract = {PurposeSublethal thermal therapy of the retinal pigment epithelium (RPE) is discussed as a new prophylactic therapy for age-related macular degeneration. However, temperature-dependent RPE cell effects have not been well elucidated. We investigated the biochemical responses of RPE cells following sublethal to lethal thermal laser irradiation. MethodsPorcine RPE cells cultured in a dish (33mm) were heated with a Thulium laser (1.92{micro}m, 1-20W, 10s) over a spot of 3mm. Temperatures during irradiation were measured with thermocouples. Cell viability was examined using annexin-V, ethidium homodimer III and Hoechst 33342 for detecting apoptotic, necrotic and living cell, respectively, by using fluorescence microscopy for localization and flow cytometry for quantification. Secretion of vascular endothelial growth factor (VEGF) for 6h following irradiation on different temperatures was assessed with Elisa assay. In order to examine a protective effect of sublethal hyperthremia, the cells were heated up to 45C 24h prior to the exposure of 2 mM hydroxyl peroxide (H2O2) for 5 h. The involvement of TRPV (Transient Receptor Potential Vanilloid)-1 receptor, which is activated with temperatures > 43C, was investigated by adding capsazepin, a TRPV-1 inhibitor, before irradiation. ResultsCell apoptosis and necrosis was observed 24 h after irradiation with a central peak temperature [&ge;]52C. Fluorescence microscopy revealed apoptotic cells around the central necrotic area. VEGF secretion for 6h after irradiation was significantly increased at peak temperatures between 40 and 52C in a temperature dependent manner (max. 110%, p<0.05), whereas the total secretion decreases with temperatures > 52C. Pre-irradiation onto 45C significantly reduced H2O2-induced cell death after 5h compared to non-heated cells (total cell death: 15.6% to 10.2%, necrosis: 6% to 4 %, early apoptosis: 5.1% to 3.6%; p<0.01). These effects were not observed in the existence of capsazepin during laser irradiation. ConclusionsThe number of apoptotic and necrotic RPE cells increase at least over 24h following thermal laser irradiation. Sublethal temperatures between 40 and 52C seem to induce various cellular responses as VEGF secretion, which might be related to the protective effect against oxidative stress. Results with capsazepin suggest that TRPV-1 channel activation by hyperthermia is essential to exert this protective effect.},
     url = {http://abstracts.iovs.org/cgi/content/abstract/54/6/1809},
     year = {2013},
     type = {Journal Article}
  }
  

2012

• Treumer, F. and Klettner, A. and Baltz, J. and Hussain, A. A. and Miura, Y. and Brinkmann, R. and Roider, J. and Hillenkamp, J.: Vectorial release of matrix metalloproteinases (MMPs) from porcine RPE-choroid explants following selective retina therapy (SRT): towards slowing the macular ageing process. Exp Eye Res, no. 97, pp. 63-72, 2012
  

  @article{Miura2012,
     author = {Treumer, F. and Klettner, A. and Baltz, J. and Hussain, A. A. and Miura, Y. and Brinkmann, R. and Roider, J. and Hillenkamp, J.},
     title = {Vectorial release of matrix metalloproteinases (MMPs) from porcine RPE-choroid explants following selective retina therapy (SRT): towards slowing the macular ageing process},
     journal = {Exp Eye Res},
     volume = {97},
     number = {1},
     pages = {63-72},
     note = {Treumer, F
  Klettner, A
  Baltz, J
  Hussain, A A
  Miura, Y
  Brinkmann, R
  Roider, J
  Hillenkamp, J
  eng
  England
  2012/03/06 06:00
  Exp Eye Res. 2012 Apr;97(1):63-72. doi: 10.1016/j.exer.2012.02.011. Epub 2012 Feb 22.},
     abstract = {The purpose of this study was to investigate release of matrix metalloproteinases (MMP) 2 and 9 during retinal pigment epithelium (RPE) wound healing after Selective Retina Therapy (SRT) with laser energy levels below and above the threshold of RPE cell death. Following exposure to SRT using a prototype pulsed Nd:YLF laser with energies of 80-180 mJ/cm(2) fresh porcine RPE-monolayers with Bruch's membrane and choroid were cultured in modified Ussing chambers which separate the apical (RPE-facing) and basal (choroid facing) sides of the RPE monolayer. Threshold energy for RPE cell death and wound healing were determined with calcein-AM viability test. Inactive and active forms of MMP 2 and 9 were quantified within tissue samples and in the culture medium of the apical and basal compartments of the Ussing chamber using gelatine zymography. Laser energies of 160-180 mJ/cm(2) resulted in cell death within 1 h while 120-140 mJ/cm(2) resulted in delayed death of exposed RPE cells. All cells survived 80 and 100 mJ/cm(2). Laser spots healed within 6 days after SRT accompanied by a transient vectorial increase of MMPs. SRT with 180 mJ/cm(2) increased active MMP 2 by 1.9 (p < 0.05) and 1.6 (p < 0.05) fold in tissue and basal compartments, respectively, without alterations in the apical compartment. Pro-MMP 2 levels were also significantly increased in all compartments (p < 0.05). Release of MMP 9 was not altered. Laser energy below the threshold of RPE cell death did not alter the release of MMP 2 or 9. The findings suggest that the release of active MMP 2 on the basal side of the RPE during wound healing following SRT may address age-related pathological changes of Bruch's membrane with a potential to slow degenerative macular ageing processes before irreversible functional loss has occurred.},
     keywords = {Animals
  Cell Death
  Cell Survival
  Choroid/*enzymology/pathology
  Diffusion Chambers, Culture
  Fluoresceins/metabolism
  *Laser Therapy
  Lasers, Solid-State
  Macular Degeneration/enzymology/pathology/*surgery
  Matrix Metalloproteinase 2/*metabolism
  Matrix Metalloproteinase 9/*metabolism
  Organ Culture Techniques
  Retinal Pigment Epithelium/*enzymology/pathology
  Sensory Thresholds
  Swine
  Wound Healing/*physiology},
     year = {2012}
  }
  

• Treumer, F. and Klettner, A. and Baltz, J. and Hussain, A. A. and Miura, Y. and Brinkmann, R. and Roider, J. and Hillenkamp, J.: Vectorial release of matrix metalloproteinases (MMPs) from porcine RPE-choroid explants following selective retina therapy (SRT): towards slowing the macular ageing process. Exp Eye Res, no. 97, pp. 63-72, 2012
  

  @article{Treumer2012,
     author = {Treumer, F. and Klettner, A. and Baltz, J. and Hussain, A. A. and Miura, Y. and Brinkmann, R. and Roider, J. and Hillenkamp, J.},
     title = {Vectorial release of matrix metalloproteinases (MMPs) from porcine RPE-choroid explants following selective retina therapy (SRT): towards slowing the macular ageing process},
     journal = {Exp Eye Res},
     volume = {97},
     number = {1},
     pages = {63-72},
     note = {1096-0007
  Treumer, F
  Klettner, A
  Baltz, J
  Hussain, A A
  Miura, Y
  Brinkmann, R
  Roider, J
  Hillenkamp, J
  Journal Article
  England
  Exp Eye Res. 2012 Apr;97(1):63-72. doi: 10.1016/j.exer.2012.02.011. Epub 2012 Feb 22.},
     abstract = {The purpose of this study was to investigate release of matrix metalloproteinases (MMP) 2 and 9 during retinal pigment epithelium (RPE) wound healing after Selective Retina Therapy (SRT) with laser energy levels below and above the threshold of RPE cell death. Following exposure to SRT using a prototype pulsed Nd:YLF laser with energies of 80-180 mJ/cm(2) fresh porcine RPE-monolayers with Bruch's membrane and choroid were cultured in modified Ussing chambers which separate the apical (RPE-facing) and basal (choroid facing) sides of the RPE monolayer. Threshold energy for RPE cell death and wound healing were determined with calcein-AM viability test. Inactive and active forms of MMP 2 and 9 were quantified within tissue samples and in the culture medium of the apical and basal compartments of the Ussing chamber using gelatine zymography. Laser energies of 160-180 mJ/cm(2) resulted in cell death within 1 h while 120-140 mJ/cm(2) resulted in delayed death of exposed RPE cells. All cells survived 80 and 100 mJ/cm(2). Laser spots healed within 6 days after SRT accompanied by a transient vectorial increase of MMPs. SRT with 180 mJ/cm(2) increased active MMP 2 by 1.9 (p < 0.05) and 1.6 (p < 0.05) fold in tissue and basal compartments, respectively, without alterations in the apical compartment. Pro-MMP 2 levels were also significantly increased in all compartments (p < 0.05). Release of MMP 9 was not altered. Laser energy below the threshold of RPE cell death did not alter the release of MMP 2 or 9. The findings suggest that the release of active MMP 2 on the basal side of the RPE during wound healing following SRT may address age-related pathological changes of Bruch's membrane with a potential to slow degenerative macular ageing processes before irreversible functional loss has occurred.},
     keywords = {Animals
  Cell Death
  Cell Survival
  Choroid/*enzymology/pathology
  Diffusion Chambers, Culture
  Fluoresceins/metabolism
  *Laser Therapy
  Lasers, Solid-State
  Macular Degeneration/enzymology/pathology/*surgery
  Matrix Metalloproteinase 2/*metabolism
  Matrix Metalloproteinase 9/*metabolism
  Organ Culture Techniques
  Retinal Pigment Epithelium/*enzymology/pathology
  Sensory Thresholds
  Swine
  Wound Healing/*physiology},
     ISSN = {0014-4835},
     DOI = {10.1016/j.exer.2012.02.011},
     year = {2012},
     type = {Journal Article}
  }
  

• Koinzer, S. and Schlott, K. and Ptaszynski, L. and Bever, M. and Kleemann, S. and Saeger, M. and Baade, A. and Caliebe, A. and Miura, Y. and Birngruber, R. and Brinkmann, R. and Roider, J.: Temperature-controlled retinal photocoagulation - a step toward automated laser treatment. Invest Ophthalmol Vis Sci, no. 53, pp. 3605-14, 2012
  

  @article{Koinzer2012,
     author = {Koinzer, S. and Schlott, K. and Ptaszynski, L. and Bever, M. and Kleemann, S. and Saeger, M. and Baade, A. and Caliebe, A. and Miura, Y. and Birngruber, R. and Brinkmann, R. and Roider, J.},
     title = {Temperature-controlled retinal photocoagulation - a step toward automated laser treatment},
     journal = {Invest Ophthalmol Vis Sci},
     volume = {53},
     number = {7},
     pages = {3605-14},
     note = {Using Smart Source Parsing
  Jun 14; Print 2012 Jul},
     abstract = {Purpose. Retinal laser photocoagulation carries the risk of overtreatment due to effect variation of identically applied lesions. The degree of coagulation depends on the induced temperature increase and on exposure time. We introduce temperature controlled photocoagulation (TCP), which uses optoacoustics to determine individually exposure times necessary to create reproducible lesions. Methods. Optoacoustic temperature measurement relies on pressure waves that are excited in the retinal tissue by repetitive low-energy laser pulses. Signal amplitudes correlate with tissue temperature and are detected by a transducer in the laser contact lens. We used a continuous wave (CW) photocoagulator for treatment irradiation and superimposed probe laser pulses for simultaneous temperature measurement. Optoacoustic data of 1500 lesions (rabbit) were evaluated to develop an algorithm that controls exposure times automatically in TCP. Lesion diameters of 156 TCP lesions were compared to 156 non-controlled lesions. Histology was performed after 1 hour, and 1 and 4 weeks. Results. TCP resulted in exposure times from 4 to 800 ms depending on laser power chosen. Ophthalmoscopic and histologic lesion diameters were independent of power between 14 and 200 mW. TCP lesions barely were visible with a mean diameter equal to the treatment beam (130 mum). In contrast, standard lesion diameters increased linearly and statistically significantly with power. Histology confirmed sparing of the ganglion and nerve fiber layers in TCP. Conclusions. TCP facilitates uniform retinal lesions over a wide power range. In a clinical setting, it should generate soft and reproducible lesions independently of local tissue variation and improve safety, particularly at short exposure times.},
     year = {2012}
  }

• Iwami, Hisashi and Ptaszynski, Lars and Danicke, Veit and Brinkmann, Ralf and Miura, Yoko: Sublethal Hyperthermia-induced Vascular Endothelial Growth Factor Secretion And Its Contribution To Adoptive Response Of Retinal Pigment Epithelial Cell. Invest. Ophthalmol. Vis. Sci., no. 53, pp. 4782-, 2012
  

  @article{Iwami2012,
     author = {Iwami, Hisashi and Ptaszynski, Lars and Danicke, Veit and Brinkmann, Ralf and Miura, Yoko},
     title = {Sublethal Hyperthermia-induced Vascular Endothelial Growth Factor Secretion And Its Contribution To Adoptive Response Of Retinal Pigment Epithelial Cell},
     journal = {Invest. Ophthalmol. Vis. Sci.},
     volume = {53},
     number = {6},
     pages = {4782-},
     abstract = {PurposeTo investigate temperature increase-induced secretion of vascular endothelial growth factor (VEGF) from retinal pigment epithelial (RPE) cells and its contribution to adoptive response relating to cell defence system against oxidative stress. MethodsPorcine RPE cells on 35 mm culture dish were used in the study. Thulium laser ({lambda}=1940 nm, spot size 33 mm was utilized as a heat source. Temperature increase during irradiation for different power and time setting at cell level was measured with thermocouple, and power and time setting of the experiment was determined based on this calibration. Culture medium was replaced by 1.2 ml phosphate buffer saline and then laser was irradiated with different power settings for 10 seconds, so that the peak temperature reaches from 40{degrees}C to 65{degrees}C. Cellular viability after laser irradiation was examined with MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay immediately after irradiation. VEGF secretion was investigated with enzyme-linked immunosorbent assay (ELISA) at 2 and 24 hrs after irradiation. Contribution of a temperature-dependent calcium channel, TRPV (transient receptor potential vanilloid) channels in laser-induced VEGF secretion was investigated using TRPV channel blocker, ruthenium red (20 {micro}M). TRPV channel blocker-containing medium was replaced by the normal medium soon after laser irradiation. Hydrogen peroxide (H2O2) or advanced glycation endproduct (AGE)-was exposed after 6 hrs of laser irradiation and cell viability was examined with MTT assay. ResultsPeak temperature threshold for immediate RPE cell death was found around 55 {degrees}C with our irradiation setting. VEGF secretion was increased after sub-lethal irradiation in power-dependent manner, which was partially suppressed by TRPV channel blocker. Sublethal laser irradiation reduced H2O2 and AGE-induced cell death and this effect was smaller in the cells treated with TRPV channel inhibitor during laser irradiation. ConclusionsSublethal temperature increase-induced VEGF production might contribute to the enhancement of RPE cell defence system against oxidative stress.},
     url = {http://abstracts.iovs.org/cgi/content/abstract/53/6/4782},
     year = {2012},
     type = {Journal Article}
  }
  

2011

• Miura, Yoko: Retinal pigment epithelium-choroid organ culture. Expert Rev Ophthalmol, no. 6, pp. 669-680, 2011
  

  @article{Miura2011,
     author = {Miura, Yoko},
     title = {Retinal pigment epithelium-choroid organ culture},
     journal = {Expert Rev  Ophthalmol},
     volume = {6},
     number = {6},
     pages = {669-680},
     abstract = {The retinal pigment epithelium (RPE) plays a vital role in retinal function, and therefore studies on RPE provide a significant benefit for our visual function. In order to obtain useful information from study results, choosing the suitable experimental model for each purpose of study is of great importance. Although RPE cell cultures are widely used, cells in cell culture have significantly different phenotypical characteristics from in vivo cells. The advantage of using native tissue is that cells in the tissue have close biological properties to in vivo conditions. This review describes basic characteristics of native RPE–choroid tissues in comparison to RPE cells in cell culture and introduces the possibility of preserving tissues in different culture systems. Advantages and disadvantages of organ culture and suitable studies with recent study results are also introduced.},
     year = {2011}
  }

• Brinkmann, R and Koinzer, S and Schlott, K and Ptaszynski, L and Bever, M and Baade, A and Miura, Y and Birngruber, R and Roider, J: Realtime temperature determination during retinal photocoagulation on patients.. Proc SPIE, no. 7885, pp. 78850R, 2011
  

  @article{Brinkmann2011,
     author = {Brinkmann, R and Koinzer, S and Schlott, K and Ptaszynski, L and Bever, M and Baade, A and Miura, Y and Birngruber, R and Roider, J},
     title = {Realtime temperature determination during retinal photocoagulation on patients.},
     journal = {Proc SPIE},
     volume = {7885},
     pages = {78850R},
     abstract = {Retinal photocoagulation is a long time established treatment for a variety of retinal diseases, most commonly applied for diabetic macular edema and diabetic retinopathy. The damage extent of the induced thermal coagulations depend on the temperature increase and the time of irradiation. So far, the induced temperature rise is unknown due to intraocular variations in light transmission and scattering and RPE/choroidal pigmentation, which can vary inter- and intraindividually by more than a factor of four. Thus in clinical practice, often stronger and deeper coagulations are applied than therapeutically needed, which lead to extended retinal damage and strong pain perception. The final goal of this project focuses on a dosimetry control, which automatically generates a desired temperature profile and thus coagulation strength for every individual coagulation spot, ideally unburden the ophthalmologist from any laser settings. In this paper we present the first realtime temperature measurements achieved on patients during retinal photocoagulation by means of an optoacoustic method, making use of the temperature dependence of the thermal expansion coefficient of retinal tissue. Therefore, nanosecond probe laser pulses are repetitively and simultaneously applied with the treatment radiation in order to excite acoustic waves, which are detected at the cornea with an ultrasonic transducer embedded in the contact lens and then are processed by PC.},
     url = {http://proceedings.spiedigitallibrary.org/proceeding.aspx?articleid=732381},
     year = {2011},
     type = {Journal Article}
  }
  

2010

• Miura, Y and Klettner, A and Roider, J: VEGF antagonists decrease barrier function of retinal pigment epithelium in vitro: possible participation of intracellular glutathione. Invest Ophthalmol Vis Sci, no. 51, pp. 4848-55, 2010
  

  @article{Miura2010,
     author = {Miura, Y and Klettner, A and Roider, J},
     title = {VEGF antagonists decrease barrier function of retinal pigment epithelium in vitro: possible participation of intracellular glutathione},
     journal = {Invest Ophthalmol Vis Sci},
     volume = {51},
     number = {9},
     pages = {4848-55},
     note = {Miura, Yoko
  Klettner, Alexa
  Roider, Johann
  United States
  Invest Ophthalmol Vis Sci. 2010 Sep;51(9):4848-55. Epub 2010 Apr 30.},
     abstract = {PURPOSE: To investigate the influence of VEGF antagonists on the barrier function of the retinal pigment epithelium and underlying mechanisms. METHODS: Porcine RPE cells were cultured on six-well membrane inserts. The cells were exposed to bevacizumab (62.5 microg/mL) or ranibizumab (25 microg/mL) for 24 hours (short term) or 9 days (long term). Transepithelial flux of FITC-dextran and intracellular levels of reduced glutathione (GSH) at normal and low-glucose conditions were investigated at different points in time. The influence of the addition of triamcinolone acetonide (TA) was investigated. The effect of GSH depletion on RPE permeability was examined using L-buthionine sulfoximine (BSO), a gamma-glutamylcysteine synthethase inhibitor. RESULTS: After short-term exposure, VEGF antagonists increased the transepithelial flux of FITC-dextran significantly on day 2. Bevacizumab, but not ranibizumab, increased permeability up to 9 days. Under long-term exposure, both drugs enhanced permeability for 7 days; bevacizumab had the stronger effect. The addition of TA inhibited this increase. At the ninth day of short- and long-term exposure, bevacizumab-exposed cells, but not ranibizumab-exposed cells, exhibited a significantly lower GSH level. In the low-glucose condition, both drugs accelerated the decrease of intracellular GSH for the first 48 hours. GSH depletion increased the permeability of retinal pigment epithelium. TA had no effect on BSO-induced GSH depletion. CONCLUSIONS: The results suggest that bevacizumab and ranibizumab may decrease RPE barrier function, with bevacizumab exhibiting a prolonged and more profound effect. Combination with TA is thought to be beneficial because of its protective effect on stabilizing RPE junctional integrity.},
     keywords = {Angiogenesis Inhibitors/ pharmacology
  Animals
  Antibodies, Monoclonal/ pharmacology
  Antibodies, Monoclonal, Humanized
  Buthionine Sulfoximine/pharmacology
  Cell Membrane Permeability/drug effects
  Cells, Cultured
  Dextrans/pharmacokinetics
  Enzyme Inhibitors/pharmacology
  Fluorescein-5-isothiocyanate/analogs & derivatives/pharmacokinetics
  Glucose/pharmacology
  Glutathione/ metabolism
  Retinal Pigment Epithelium/cytology/ drug effects/ metabolism
  Swine
  Tight Junctions/drug effects/metabolism
  Vascular Endothelial Growth Factor A/ antagonists & inhibitors/metabolis
  AutoPhoN},
     ISSN = {1552-5783 (Electronic)
  0146-0404 (Linking)},
     DOI = {10.1167/iovs.09-4699},
     year = {2010},
     type = {Journal Article}
  }
  

• Miura, Y. and Klettner, A. and Noelle, B. and Hasselbach, H. and Roider, J.: Change of morphological and functional characteristics of retinal pigment epithelium cells during cultivation of retinal pigment epithelium-choroid perfusion tissue culture. Ophthalmic Res, no. 43, pp. 122-33, 2010
  

  @article{Miura2009,
     author = {Miura, Y. and Klettner, A. and Noelle, B. and Hasselbach, H. and Roider, J.},
     title = {Change of morphological and functional characteristics of retinal pigment epithelium cells during cultivation of retinal pigment epithelium-choroid perfusion tissue culture},
     journal = {Ophthalmic Res},
     volume = {43},
     number = {3},
     pages = {122-33},
     note = {Using Smart Source Parsing
  Epub 2009 Oct 29},
     abstract = {AIMS: To evaluate the changes of morphological and functional characteristics of the retinal pigment epithelium (RPE)-choroid perfusion culture during cultivation. METHODS: PorcineRPE-choroid tissue was cultivated in a perfusion tissue culture system. After the indicated times, histology, immunolocalization of collagen IV and von Willebrand factor, RPE cell viability with calcein-AM, TUNEL assay and occludin immunolocalization of RPE cells were examined. The tissue was treated with selective RPE treatment laser after different time periods and the wound healing response was characterized. Vascular endothelial growth factor secretion was measured by enzyme-linked immunosorbent assay. RESULTS: On day 8, prominent morphological degenerative changes of RPE cells were observed in histology. According to the immunohistochemistry for collagen IV, the Bruch's membrane did not display any obvious decomposition until day 8. Von Willebrand factor staining decreased during cultivation, especially at the choriocapillaris. Calcein-AM staining and TUNEL assay displayed the increase of apoptotic changes in only a minority of the cells on day 4, but in many cells on day 8. Occludin delocalization was observed on day 8. Selective RPE treatment laser-produced wounds were completely closed by monolayer RPE when wounded on fresh and 3-day-old cultures, but not when wounded on 6-day-old cultures. Vascular endothelial growth factor secretion was stable between days 2 and 5, but increased after that. CONCLUSION: Under the stated culture perfusion conditions, porcine RPE-choroid tissue was suitable for experimentation up to 5 days of maintenance.},
     year = {2010}
  }

• Miura, Y. and Klettner, A. and Noelle, B. and Hasselbach, H. and Roider, J.: Change of morphological and functional characteristics of retinal pigment epithelium cells during cultivation of retinal pigment epithelium-choroid perfusion tissue culture. Ophthalmic Res, no. 43, pp. 122-33, 2010
  

  @article{Miura2009,
     author = {Miura, Y. and Klettner, A. and Noelle, B. and Hasselbach, H. and Roider, J.},
     title = {Change of morphological and functional characteristics of retinal pigment epithelium cells during cultivation of retinal pigment epithelium-choroid perfusion tissue culture},
     journal = {Ophthalmic Res},
     volume = {43},
     number = {3},
     pages = {122-33},
     note = {1423-0259
  Miura, Yoko
  Klettner, Alexa
  Noelle, Bernhard
  Hasselbach, Heike
  Roider, Johann
  Journal Article
  Switzerland
  Ophthalmic Res. 2010;43(3):122-33. doi: 10.1159/000252979. Epub 2009 Oct 29.},
     abstract = {AIMS: To evaluate the changes of morphological and functional characteristics of the retinal pigment epithelium (RPE)-choroid perfusion culture during cultivation. METHODS: PorcineRPE-choroid tissue was cultivated in a perfusion tissue culture system. After the indicated times, histology, immunolocalization of collagen IV and von Willebrand factor, RPE cell viability with calcein-AM, TUNEL assay and occludin immunolocalization of RPE cells were examined. The tissue was treated with selective RPE treatment laser after different time periods and the wound healing response was characterized. Vascular endothelial growth factor secretion was measured by enzyme-linked immunosorbent assay. RESULTS: On day 8, prominent morphological degenerative changes of RPE cells were observed in histology. According to the immunohistochemistry for collagen IV, the Bruch's membrane did not display any obvious decomposition until day 8. Von Willebrand factor staining decreased during cultivation, especially at the choriocapillaris. Calcein-AM staining and TUNEL assay displayed the increase of apoptotic changes in only a minority of the cells on day 4, but in many cells on day 8. Occludin delocalization was observed on day 8. Selective RPE treatment laser-produced wounds were completely closed by monolayer RPE when wounded on fresh and 3-day-old cultures, but not when wounded on 6-day-old cultures. Vascular endothelial growth factor secretion was stable between days 2 and 5, but increased after that. CONCLUSION: Under the stated culture perfusion conditions, porcine RPE-choroid tissue was suitable for experimentation up to 5 days of maintenance.},
     keywords = {Animals
  *Apoptosis
  Bruch Membrane/pathology
  Cell Survival
  Choroid/metabolism/*pathology/surgery
  Collagen Type IV/metabolism
  Enzyme-Linked Immunosorbent Assay
  Fluoresceins/metabolism
  Immunoenzyme Techniques
  In Situ Nick-End Labeling
  Laser Therapy
  Membrane Proteins/metabolism
  Occludin
  Organ Culture Techniques
  Retinal Pigment Epithelium/metabolism/*pathology/surgery
  Swine
  Time Factors
  Vascular Endothelial Growth Factor A/metabolism
  Wound Healing
  von Willebrand Factor/metabolism},
     ISSN = {0030-3747},
     DOI = {10.1159/000252979},
     year = {2010},
     type = {Journal Article}
  }
  

2009

• Miura, Y. and Roider, J.: Triamcinolone acetonide prevents oxidative stress-induced tight junction disruption of retinal pigment epithelial cells. Graefes Arch Clin Exp Ophthalmol, no. 247, pp. 641-9, 2009
  

  @article{Miura2009,
     author = {Miura, Y. and Roider, J.},
     title = {Triamcinolone acetonide prevents oxidative stress-induced tight junction disruption of retinal pigment epithelial cells},
     journal = {Graefes Arch Clin Exp Ophthalmol},
     volume = {247},
     number = {5},
     pages = {641-9},
     note = {Using Smart Source Parsing
  May; Epub 2009 Feb 3},
     abstract = {PURPOSE: Oxidative stress is known to disrupt the integrity of retinal pigment epithelium (RPE) tight junctions. The goal of this study is to evaluate the effect of triamcinolone acetonide (TA) on the junctional integrity of RPE under oxidative stress and to identify the underlying mechanisms. METHODS: Second passage porcine RPE cells were cultured on 6-well membrane inserts until 4 weeks after reaching confluence. Cells were incubated with TA (10(-5) M) for 30 min. FITC-containing medium was added to the upper chamber (cell's apical side). The cells were then challenged with 1 mM Hydrogen Peroxide (H(2)O(2)). After 5 h, the fluorescence intensity of the medium from lower chamber (cell's basolateral side) was measured using a fluorescence spectrofluorophotometer. This transepithelial flux of FITC-dextran was measured until the 21st day. The immunolocalization of occludin and F-actin was examined with fluorescence microscope. Reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio was determined by a colorimetric assay kit. RESULTS: Non-lethal oxidative stress by H(2)O(2) increased transepithelial flux of FITC-dextran significantly. TA inhibited this increase and preserved the lower flux through the whole experimental period. This permeability change by H(2)O(2) was reversible and recovered to the normal level within 3 weeks. In immunohistological study, H(2)O(2) reduced linear occludin staining at the cell border and increased actin stress fibers. TA prevented H(2)O(2)-induced disruption of junctional assembly of occludin and F-actin. Glutathione assay demonstrated that intracellular GSH/GSSG ratio decreased significantly with H(2)O(2), while TA preserved this ratio by up-regulating GSH synthesis. CONCLUSIONS: TA has a protective effect against oxidative stress-induced disruption of RPE tight junction by preserving cellular redox state.},
     year = {2009}
  }