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Incipient UV-Induced Structural Changes in Neutrophil Granulocytes: Morphometric and Texture Analysis of Two-Dimensional Digital Images

Published online by Cambridge University Press:  24 February 2016

Ivan Grbatinić  and
Nebojša T. Milošević
Ivan Grbatinić*
Affiliation:
Laboratory of Digital Image Processing, School of Medicine, University of Belgrade, Visegradka 2, Belgrade, Serbia
Nebojša T. Milošević
Affiliation:
Department of Biophysics, School of Medicine, University of Belgrade, Visegradka 2, Belgrade, Serbia
*
*Corresponding author. igrbson@gmail.com
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Abstract

The aim of this study is to determine the ability and consequent significance of fractal and lacunarity analysis together with computational morphometric and gray-level co-occurrence matrix (GLCM) analysis in detecting subtle initial UVB-induced chromatin and cytosolic changes in neutrophil granulocytes. In addition, the direction and potential significance of the observed changes is speculated. Feulgen-stained neutrophils are pictured and their digitalized images are analyzed in specialized software for digital image processing and ImageJ analysis. Significant statistical difference is observed (p<0.01) between UV-treated neutrophils and controls in the vast majority of cytoplasmic morphometric parameters. Nuclear GLCM analysis was very sensitive with just GLCM homogeneity as non-significant between UV-treated cells and controls (p>0.05). For other parameters there was mostly high statistical significance (p>0.05). Significant unmatched correlations were found as sensitive markers of early morphological changes in cells exposed to UV light. In addition, the correlation between nuclear area and entropy was determined and was highly significant (p<0.001). UVB light, due to its high absorbance by DNA molecules, leads to double behavior of the cells. On one hand, cells start to rearrange but on the other UV light starts very early to immediately damage the cell. All these processes are very subtle in their intensity and GLCM analysis and computational imaging methods based on fractal geometry, i.e. fractal and morphometric analysis, in particular their combination, are very sensitive for detecting and describing these early chromatin changes.

Keywords

neutrophil granulocytesfractal analysislacunarity analysiscomputational morphometric analysisgray-level co-occurrence matrix analysis
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 22 , Issue 2 , April 2016 , pp. 387 - 393
Copyright
© Microscopy Society of America 2016 

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References

Arita, K., Yamamoto, Y., Takehara, Y., Utsumi, T., Kanno, T., Miyaguchi, C., Akiyama, J., Yoshioka, T. & Utsumi, K. (2003). Mechanisms of enhanced apoptosis in HL-60 cells by UV-irradiated n-3 and n-6 polyunsaturated fatty acids. Free Radic Biol Med 35, 189–199.CrossRefGoogle ScholarPubMed
Assefa, D., Keller, H., Menard, C., Laperriere, N., Ferrari, R.J. & Yeung, I. (2010). Robust texture features for response monitoring of glioblastoma multiforme on T1-weighted and T2-FLAIR MR images: A preliminary investigation in terms of identification and segmentation. Med Phys 37, 1722–1736.Google Scholar
Bancaud, A., Huet, S., Daigle, N., Mozziconacci, J., Beaudouin, J. & Ellenberg, J. (2009). Molecular crowding affects diffusion and binding of nuclear proteins in heterochromatin and reveals the fractal organization of chromatin. EMBO J 28, 3785–3798.CrossRefGoogle ScholarPubMed
Batista, L.F., Kaina, B., Meneghini, R. & Menck, C.F. (2009). How DNA lesions are turned into powerful killing structures: Insights from UV-induced apoptosis. Mutat Res 681, 197–208.CrossRefGoogle ScholarPubMed
Bizzarri, M., Giuliani, A., Cucina, A., D’Anselmi, F., Soto, A.M. & Sonnenschein, C. (2011). Fractal analysis in a systems biology approach to cancer. Semin Cancer Biol 21, 175–182.CrossRefGoogle Scholar
Cabrera, J. (2005). TextureAnalyzer. Available at http://rbsinfonihgov/ij/plugins/texturehtml.Google Scholar
Cadet, J., Grand, A. & Douki, T. (2015). Solar UV radiation-induced DNA bipyrimidine photoproducts: Formation and mechanistic insights. Top Curr Chem 356, 249–275.Google Scholar
Castelli, C. & Losa, G.A. (2001). Ultrastructural complexity of nuclear components during early apoptotic phases in breast cancer cells. Anal Cell Pathol 23, 1–9.Google Scholar
Chan, H.P., Wu, Y.T., Sahiner, B., Wei, J., Helvie, M.A., Zhang, Y., Moore, R.H., Kopans, D.B., Hadjiiski, L. & Way, T. (2010). Characterization of masses in digital breast tomosynthesis: Comparison of machine learning in projection views and reconstructed slices. Med Phys 37, 3576–3586.Google Scholar
Cornish, T.B. (2007). GLCMTextureToo. Available at http://tobycornishcom/downloads/imagej.Google Scholar
D’Anselmi, F., Valerio, M., Cucina, A., Galli, L., Proietti, S., Dinicola, S., Pasqualato, A., Manetti, C., Ricci, G., Giuliani, A. & Bizzarri, M. (2011). Metabolism and cell shape in cancer: A fractal analysis. Int J Biochem Cell Biol 43, 1052–1058.Google Scholar
Dokukin, M.E., Guz, N.V., Gaikwad, R.M., Woodworth, C.D. & Sokolov, I. (2011). Cell surface as a fractal: Normal and cancerous cervical cells demonstrate different fractal behavior of surface adhesion maps at the nanoscale. Phys Rev Lett 107, 028101.Google Scholar
Ehrhart, J.C., Gosselet, F.P., Culerrier, R.M. & Sarasin, A. (2003). UVB-induced mutations in human key gatekeeper genes governing signalling pathways and consequences for skin tumourigenesis. Photochem Photobiol Sci 2, 825–834.CrossRefGoogle ScholarPubMed
Farrell, A.W., Halliday, G.M. & Lyons, J.G. (2011). Chromatin structure following UV-induced DNA damage—repair or death? Int J Mol Sci 12, 8063–8085.Google Scholar
Gill, S.S., Anjum, N.A., Gill, R., Jha, M. & Tuteja, N. (2015). DNA damage and repair in plants under ultraviolet and ionizing radiations. Scientific World Journal 2015, 250158.Google Scholar
Haralick, R.M., Shanmugam, K. & Dinstein, I. (1973). Textural features for image classification. IEEE Trans Syst Man Cybern 3, 610–621.CrossRefGoogle Scholar
Huber, M.B., Nagarajan, M.B., Leinsinger, G., Eibel, R., Ray, L.A. & Wismüller, A. (2011). Performance of topological texture features to classify fibrotic interstitial lung disease patterns. Med Phys 38, 2035–2044.Google Scholar
Jambawalikar, S., Li, H., Shah, S., Fisher, P. & Button, T. (2011). SU-E-I-04: Texture feature based CAD for breast cancer detection. Med Phys 38, 3396.Google Scholar
Kao, E.F., Kuo, Y.T., Hsu, J.S., Chou, M.C. & Liu, G.C. (2011). Zone-based analysis for automated detection of abnormalities in chest radiographs. Med Phys 38, 4241–4250.CrossRefGoogle ScholarPubMed
Karperien, A. (1999–2014). FracLacforImageJ. V.2.5.Google Scholar
Kulms, D. & Schwarz, T. (2000). Molecular mechanisms of UV-induced apoptosis. Photodermatol Photoimmunol Photomed 16, 195–201.Google Scholar
Kuo, H.R., Lapidus, A. & Lambert, W.C. (1998). Identification of early apoptosis in Feulgen-stained cultured cells in situ by computerized image analysis. Cytometry 33, 420–427.3.0.CO;2-F>CrossRefGoogle ScholarPubMed
Li, H., Liu, L., Xing, D. & Chen, W.R. (2010). Inhibition of the JNK/Bim pathway by Hsp70 prevents Bax activation in UV-induced apoptosis. FEBS Lett 584, 4672–4678.CrossRefGoogle ScholarPubMed
Liao, Y.Y., Tsui, P.H., Li, C.H., Chang, K.J., Kuo, W.H., Chang, C.C. & Yeh, C.K. (2011). Classification of scattering media within benign and malignant breast tumors based on ultrasound texture-feature-based and Nakagami-parameter images. Med Phys 38, 2198–2207.CrossRefGoogle ScholarPubMed
Lopes, R. & Betrouni, N. (2009). Fractal and multifractal analysis: A review. Med Image Anal 13, 634–649.Google Scholar
Losa, G.A. & Castelli, C. (2005). Nuclear patterns of human breast cancer cells during apoptosis: Characterisation by fractal dimension and co-occurrence matrix statistics. Cell Tissue Res 322, 257–267.Google Scholar
Mancebo, S.E. & Wang, S.Q. (2014). Skin cancer: Role of ultraviolet radiation in carcinogenesis. Rev Environ Health 29, 265–273.CrossRefGoogle ScholarPubMed
Palomera-Sanchez, Z. & Zurita, M. (2011). Open, repair and close again: Chromatin dynamics and the response to UV-induced DNA damage. DNA Repair (Amst) 10, 119–125.CrossRefGoogle ScholarPubMed
Pantic, I., Dacic, S., Brkic, P., Lavrnja, I., Jovanovic, T., Pantic, S. & Pekovic, S. (2015). Discriminatory ability of fractal and grey level co-occurrence matrix methods in structural analysis of hippocampus layers. J Theor Biol 370, 151–156.Google Scholar
Pantic, I., Harhaji-Trajkovic, L., Pantovic, A., Milosevic, N.T. & Trajkovic, V. (2012 a). Changes in fractal dimension and lacunarity as early markers of UV-induced apoptosis. J Theor Biol 303, 87–92.Google Scholar
Pantic, I., Pantic, S. & Basta-Jovanovic, G. (2012 b). Gray level co-occurrence matrix texture analysis of germinal center light zone lymphocyte nuclei: Physiology viewpoint with focus on apoptosis. Microsc Microanal 18, 470–475.Google Scholar
Ranjanomennahary, P., Ghalila, S.S., Malouche, D., Marchadier, A., Rachidi, M., Benhamou, C. & Chappard, C. (2011). Comparison of radiograph-based texture analysis and bone mineral density with three-dimensional microarchitecture of trabecular bone. Med Phys 38, 420–428.Google Scholar
Rass, K. & Reichrath, J. (2008). UV damage and DNA repair in malignant melanoma and nonmelanoma skin cancer. Adv Exp Med Biol 624, 162–178.CrossRefGoogle ScholarPubMed
Reichrath, J. & Rass, K. (2014). Ultraviolet damage, DNA repair and vitamin D in nonmelanoma skin cancer and in malignant melanoma: An update. Adv Exp Med Biol 810, 208–233.Google Scholar
Takemura, T., Shinomiya, N. & Rokutanda, M. (1999). Role of G1 phase in the UV-induced apoptosis of EL-4 mouse lymphoma cells. Apoptosis 4, 245–253.CrossRefGoogle ScholarPubMed
Tremblay, M., Toussaint, M., D’amours, A. & Conconi, A. (2009). Nucleotide excision repair and photolyase repair of UV photoproducts in nucleosomes: Assessing the existence of nucleosome and non-nucleosome rDNA chromatin in vivo. Biochem Cell Biol 87, 337–346.Google Scholar
Uppal, S.O., Voronine, D.V., Wendt, E. & Heckman, C.A. (2010). Morphological fractal analysis of shape in cancer cells treated with combinations of microtubule-polymerizing and -depolymerizing agents. Microsc Microanal 16, 472–477.CrossRefGoogle ScholarPubMed
Wang, X., Becker, F.F. & Gascoyne, P.R. (2010). The fractal dimension of cell membrane correlates with its capacitance: A new fractal single-shell model. Chaos 20, 043133.Google Scholar
Waugh, S.A., Lerski, R.A., Bidaut, L. & Thompson, A.M. (2011). The influence of field strength and different clinical breast MRI protocols on the outcome of texture analysis using foam phantoms. Med Phys 38, 5058–5066.CrossRefGoogle ScholarPubMed
Ying, S. & Hacker, G. (2007). Apoptosis induced by direct triggering of mitochondrial apoptosis proceeds in the near-absence of some apoptotic markers. Apoptosis 12, 2003–2011.Google Scholar
Yokoyama, H. & Mizutani, R. (2014). Structural biology of DNA (6–4) photoproducts formed by ultraviolet radiation and interactions with their binding proteins. Int J Mol Sci 15, 20321–20338.CrossRefGoogle ScholarPubMed
Zhang, Y., Wang, J.S., Chen, L.L., Cheng, X.K., Heng, F.Y., Wu, N.H. & Shen, Y.F. (2004). Repression of hsp90beta gene by p53 in UV irradiation-induced apoptosis of Jurkat cells. J Biol Chem 279, 42545–42551.CrossRefGoogle ScholarPubMed