![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 12, Issue 1, 27-36, January 2001
1 Mediates Epithelial to
Mesenchymal Transdifferentiation through a RhoA-dependent Mechanism
Vanderbilt-Ingram Cancer Center, Departments of Cancer Biology and
Medicine, Vanderbilt University Medical Center, Nashville, Tennessee
37232
Transforming growth factor-
1 (TGF-) can be tumor suppressive,
but it can also enhance tumor progression by stimulating the complex
process of epithelial-to-mesenchymal transdifferentiaion (EMT). The
signaling pathway(s) that regulate EMT in response to TGF- are not
well understood. We demonstrate the acquisition of a fibroblastoid
morphology, increased N-cadherin expression, loss of junctional
E-cadherin localization, and increased cellular motility as markers for
TGF--induced EMT. The expression of a dominant-negative Smad3 or
the expression of Smad7 to levels that block growth inhibition and
transcriptional responses to TGF- do not inhibit mesenchymal
differentiation of mammary epithelial cells. In contrast, we show that
TGF- rapidly activates RhoA in epithelial cells, and that blocking
RhoA or its downstream target p160ROCK, by the expression
of dominant-negative mutants, inhibited TGF--mediated EMT. The data
suggest that TGF- rapidly activates RhoA-dependent signaling
pathways to induce stress fiber formation and mesenchymal characteristics.
This article has been cited by other articles:
|
|
 |
|
  V. R. Placencio, A.-R. Sharif-Afshar, X. Li, H. Huang, C. Uwamariya, E. G. Neilson, M. M. Shen, R. J. Matusik, S. W. Hayward, and N. A. Bhowmick Stromal Transforming Growth Factor-{beta} Signaling Mediates Prostatic Response to Androgen Ablation by Paracrine Wnt Activity Cancer Res., June 15, 2008; 68(12): 4709 - 4718. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. M. Bailey and J. Liu Caveolin-1 Up-regulation during Epithelial to Mesenchymal Transition Is Mediated by Focal Adhesion Kinase J. Biol. Chem., May 16, 2008; 283(20): 13714 - 13724. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. A. Townsend, J. L. Wrana, G. E. Davis, and J. V. Barnett Transforming Growth Factor-{beta}-stimulated Endocardial Cell Transformation Is Dependent on Par6c Regulation of RhoA J. Biol. Chem., May 16, 2008; 283(20): 13834 - 13841. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. J. Northey, J. Chmielecki, E. Ngan, C. Russo, M. G. Annis, W. J. Muller, and P. M. Siegel Signaling through ShcA Is Required for Transforming Growth Factor {beta}- and Neu/ErbB-2-Induced Breast Cancer Cell Motility and Invasion Mol. Cell. Biol., May 15, 2008; 28(10): 3162 - 3176. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Sebe, S.-K. Leivonen, A. Fintha, A. Masszi, L. Rosivall, V.-M. Kahari, and I. Mucsi Transforming growth factor-{beta}-induced alpha-smooth muscle cell actin expression in renal proximal tubular cells is regulated by p38{beta} mitogen-activated protein kinase, extracellular signal-regulated protein kinase1,2 and the Smad signalling during epithelial-myofibroblast transdifferentiation Nephrol. Dial. Transplant., May 1, 2008; 23(5): 1537 - 1545. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. S. Acharya, S. Majumdar, M. Jacob, J. Hayden, P. Mrass, W. Weninger, R. K. Assoian, and E. Pure Fibroblast migration is mediated by CD44-dependent TGF{beta} activation J. Cell Sci., May 1, 2008; 121(9): 1393 - 1402. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. I. Kim, J. H. Kwak, L. Wang, and M. E. Choi Protein Phosphatase 2A Is a Negative Regulator of Transforming Growth Factor-{beta}1-induced TAK1 Activation in Mesangial Cells J. Biol. Chem., April 18, 2008; 283(16): 10753 - 10763. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Yin, P. Castagnino, and R. K. Assoian ABCG2 Expression and Side Population Abundance Regulated by a Transforming Growth Factor {beta}-Directed Epithelial-Mesenchymal Transition Cancer Res., February 1, 2008; 68(3): 800 - 807. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. J. Galliher-Beckley and W. P. Schiemann Grb2 binding to Tyr284 in T{beta}R-II is essential for mammary tumor growth and metastasis stimulated by TGF-{beta} Carcinogenesis, February 1, 2008; 29(2): 244 - 251. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Morita, T. Mayanagi, and K. Sobue Dual roles of myocardin-related transcription factors in epithelial mesenchymal transition via slug induction and actin remodeling J. Cell Biol., December 3, 2007; 179(5): 1027 - 1042. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Pandey, S. M. Umphress, Y. Kang, J. Angdisen, A. Naumova, K. L. Mercer, T. Jacks, and S. B. Jakowlew Modulation of tumor induction and progression of oncogenic K-ras-positive tumors in the presence of TGF- 1 haploinsufficiency Carcinogenesis, December 1, 2007; 28(12): 2589 - 2596. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Schunke, P. Span, H. Ronneburg, A. Dittmer, M. Vetter, H.-J. Holzhausen, E. Kantelhardt, S. Krenkel, V. Muller, F. C.G.J. Sweep, et al. Cyclooxygenase-2 Is a Target Gene of Rho GDP Dissociation Inhibitor {beta} in Breast Cancer Cells Cancer Res., November 15, 2007; 67(22): 10694 - 10702. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. L. Zandy, M. Playford, and A. M. Pendergast Abl tyrosine kinases regulate cell cell adhesion through Rho GTPases PNAS, November 6, 2007; 104(45): 17686 - 17691. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Beisswenger, C. B. Coyne, M. Shchepetov, and J. N. Weiser Role of p38 MAP Kinase and Transforming Growth Factor-beta Signaling in Transepithelial Migration of Invasive Bacterial Pathogens J. Biol. Chem., September 28, 2007; 282(39): 28700 - 28708. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. C. Willis and Z. Borok TGF-beta-induced EMT: mechanisms and implications for fibrotic lung disease Am J Physiol Lung Cell Mol Physiol, September 1, 2007; 293(3): L525 - L534. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Shirakihara, M. Saitoh, and K. Miyazono Differential Regulation of Epithelial and Mesenchymal Markers by {delta}EF1 Proteins in Epithelial Mesenchymal Transition Induced by TGF-beta Mol. Biol. Cell, September 1, 2007; 18(9): 3533 - 3544. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Zeisberg, C. Yang, M. Martino, M. B. Duncan, F. Rieder, H. Tanjore, and R. Kalluri Fibroblasts Derive from Hepatocytes in Liver Fibrosis via Epithelial to Mesenchymal Transition J. Biol. Chem., August 10, 2007; 282(32): 23337 - 23347. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Wang, M. Zheng, G. Liu, W. Xia, P. J. McKeown-Longo, M.-C. Hung, and J. Zhao Kruppel-Like Factor 8 Induces Epithelial to Mesenchymal Transition and Epithelial Cell Invasion Cancer Res., August 1, 2007; 67(15): 7184 - 7193. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Lamouille and R. Derynck Cell size and invasion in TGF-{beta} induced epithelial to mesenchymal transition is regulated by activation of the mTOR pathway J. Cell Biol., July 24, 2007; 178(3): 437 - 451. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Sonoyama, B.-M. Seo, T. Yamaza, and S. Shi Human Hertwig's Epithelial Root Sheath Cells Play Crucial Roles in Cementum Formation J. Dent. Res., July 1, 2007; 86(7): 594 - 599. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Arciniegas, M. G. Frid, I. S. Douglas, and K. R. Stenmark Perspectives on endothelial-to-mesenchymal transition: potential contribution to vascular remodeling in chronic pulmonary hypertension Am J Physiol Lung Cell Mol Physiol, July 1, 2007; 293(1): L1 - L8. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Xie, M. B. Sukkar, R. Issa, N. M. Khorasani, and K. F. Chung Mechanisms of induction of airway smooth muscle hyperplasia by transforming growth factor-beta Am J Physiol Lung Cell Mol Physiol, July 1, 2007; 293(1): L245 - L253. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Vyas-Read, P. W. Shaul, I. S. Yuhanna, and B. C. Willis Nitric oxide attenuates epithelial-mesenchymal transition in alveolar epithelial cells Am J Physiol Lung Cell Mol Physiol, July 1, 2007; 293(1): L212 - L221. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Choi, S. Y. Park, and C.-K. Joo Transforming Growth Factor-{beta}1 Represses E-Cadherin Production via Slug Expression in Lens Epithelial Cells Invest. Ophthalmol. Vis. Sci., June 1, 2007; 48(6): 2708 - 2718. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J. Galliher and W. P. Schiemann Src Phosphorylates Tyr284 in TGF-{beta} Type II Receptor and Regulates TGF-{beta} Stimulation of p38 MAPK during Breast Cancer Cell Proliferation and Invasion Cancer Res., April 15, 2007; 67(8): 3752 - 3758. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J. LaGier, S. H. Yoo, E. C. Alfonso, S. Meiners, and M. E. Fini Inhibition of Human Corneal Epithelial Production of Fibrotic Mediator TGF-{beta}2 by Basement Membrane-Like Extracellular Matrix Invest. Ophthalmol. Vis. Sci., March 1, 2007; 48(3): 1061 - 1071. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Zhu, A. R. Krakowski, E. E. Dunham, L. Wang, A. Bandyopadhyay, R. Berdeaux, G. S. Martin, L. Sun, and K. Luo Dual Role of SnoN in Mammalian Tumorigenesis Mol. Cell. Biol., January 1, 2007; 27(1): 324 - 339. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Seoane Escaping from the TGF{beta} anti-proliferative control Carcinogenesis, November 1, 2006; 27(11): 2148 - 2156. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Martin-Villar, D. Megias, S. Castel, M. M. Yurrita, S. Vilaro, and M. Quintanilla Podoplanin binds ERM proteins to activate RhoA and promote epithelial-mesenchymal transition J. Cell Sci., November 1, 2006; 119(21): 4541 - 4553. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Fu, F. Liu, S. Su, W. Wang, X. R. Huang, M. L. Entman, R. J. Schwartz, L. Wei, and H. Y. Lan Signaling Mechanism of Renal Fibrosis in Unilateral Ureteral Obstructive Kidney Disease in ROCK1 Knockout Mice J. Am. Soc. Nephrol., November 1, 2006; 17(11): 3105 - 3114. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. D. Andl, B. B. Fargnoli, T. Okawa, M. Bowser, M. Takaoka, H. Nakagawa, A. Klein-Szanto, X. Hua, M. Herlyn, and A. K. Rustgi Coordinated Functions of E-Cadherin and Transforming Growth Factor {beta} Receptor II In vitro and In vivo. Cancer Res., October 15, 2006; 66(20): 9878 - 9885. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. E. Wang, I. Shin, F. Y. Wu, D. B. Friedman, and C. L. Arteaga HER2/Neu (ErbB2) Signaling to Rac1-Pak1 Is Temporally and Spatially Modulated by Transforming Growth Factor {beta} Cancer Res., October 1, 2006; 66(19): 9591 - 9600. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-S. Kim, J.-G. Kim, M.-Y. Moon, C.-Y. Jeon, H.-Y. Won, H.-J. Kim, Y.-J. Jeon, J.-Y. Seo, J.-I. Kim, J. Kim, et al. Transforming growth factor-beta1 regulates macrophage migration via RhoA Blood, September 15, 2006; 108(6): 1821 - 1829. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. M. Zhao and F. M. Hoffmann Inhibition of Transforming Growth Factor-beta1-induced Signaling and Epithelial-to-Mesenchymal Transition by the Smad-binding Peptide Aptamer Trx-SARA Mol. Biol. Cell, September 1, 2006; 17(9): 3819 - 3831. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Chen, B. M. Necela, W. Su, M. Yanagisawa, P. Z. Anastasiadis, A. P. Fields, and E. A. Thompson Peroxisome Proliferator-activated Receptor {gamma} Promotes Epithelial to Mesenchymal Transformation by Rho GTPase-dependent Activation of ERK1/2 J. Biol. Chem., August 25, 2006; 281(34): 24575 - 24587. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Ao, K. Williams, N. A. Bhowmick, and S. W. Hayward Transforming Growth Factor-{beta} Promotes Invasion in Tumorigenic but not in Nontumorigenic Human Prostatic Epithelial Cells Cancer Res., August 15, 2006; 66(16): 8007 - 8016. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Lu, E. O. Harrington, H. Jackson, N. Morin, C. Shannon, and S. Rounds Transforming growth factor-beta1-induced endothelial barrier dysfunction involves Smad2-dependent p38 activation and subsequent RhoA activation J Appl Physiol, August 1, 2006; 101(2): 375 - 384. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G.-P. Sun, M. Kohno, P. Guo, Y. Nagai, K. Miyata, Y.-Y. Fan, S. Kimura, H. Kiyomoto, K. Ohmori, D.-T. Li, et al. Involvements of Rho-Kinase and TGF-beta Pathways in Aldosterone-Induced Renal Injury J. Am. Soc. Nephrol., August 1, 2006; 17(8): 2193 - 2201. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H.-J. Park, S. M. Ward, J. S. Desgrosellier, S. P. Georgescu, A. G. Papageorge, X. Zhuang, J. V. Barnett, and J. B. Galper Transforming Growth Factor beta Regulates the Expression of the M2 Muscarinic Receptor in Atrial Myocytes via an Effect on RhoA and p190RhoGAP J. Biol. Chem., July 21, 2006; 281(29): 19995 - 20002. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Varon, F. Tatin, V. Moreau, E. Van Obberghen-Schilling, S. Fernandez-Sauze, E. Reuzeau, I. Kramer, and E. Genot Transforming Growth Factor {beta} Induces Rosettes of Podosomes in Primary Aortic Endothelial Cells. Mol. Cell. Biol., May 1, 2006; 26(9): 3582 - 3594. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. R. Alexander, N. L. Tran, H. Rekapally, C. E. Summers, C. Glackin, and R. L. Heimark N-cadherin Gene Expression in Prostate Carcinoma Is Modulated by Integrin-Dependent Nuclear Translocation of Twist1. Cancer Res., April 1, 2006; 66(7): 3365 - 3369. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Medici, E. D. Hay, and D. A. Goodenough Cooperation between Snail and LEF-1 Transcription Factors Is Essential for TGF-beta1-induced Epithelial-Mesenchymal Transition Mol. Biol. Cell, April 1, 2006; 17(4): 1871 - 1879. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Deckers, M. van Dinther, J. Buijs, I. Que, C. Lowik, G. van der Pluijm, and P. ten Dijke The Tumor Suppressor Smad4 Is Required for Transforming Growth Factor {beta}-Induced Epithelial to Mesenchymal Transition and Bone Metastasis of Breast Cancer Cells Cancer Res., February 15, 2006; 66(4): 2202 - 2209. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Chen, M. Crawford, R. M. Day, V. R. Briones, J. E. Leader, P. A. Jose, and R. J. Lechleider RhoA Modulates Smad Signaling during Transforming Growth Factor-beta-induced Smooth Muscle Differentiation J. Biol. Chem., January 20, 2006; 281(3): 1765 - 1770. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Maeda, Y. Shintani, M. J. Wheelock, and K. R. Johnson Src Activation Is Not Necessary for Transforming Growth Factor (TGF)-{beta}-mediated Epithelial to Mesenchymal Transitions (EMT) in Mammary Epithelial Cells: PP1 DIRECTLY INHIBITS TGF-{beta} RECEPTORS I AND II J. Biol. Chem., January 6, 2006; 281(1): 59 - 68. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. C. Willis, R. M. duBois, and Z. Borok Epithelial Origin of Myofibroblasts during Fibrosis in the Lung. Proceedings of the ATS, January 1, 2006; 3(4): 377 - 382. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Sotgia, T. M. Williams, W. Schubert, F. Medina, C. Minetti, R. G. Pestell, and M. P. Lisanti Caveolin-1 Deficiency (-/-) Conveys Premalignant Alterations in Mammary Epithelia, with Abnormal Lumen Formation, Growth Factor Independence, and Cell Invasiveness Am. J. Pathol., January 1, 2006; 168(1): 292 - 309. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Saika, K. Ikeda, O. Yamanaka, K. C. Flanders, Y. Ohnishi, Y. Nakajima, Y. Muragaki, and A. Ooshima Adenoviral gene transfer of BMP-7, Id2, or Id3 suppresses injury-induced epithelial-to-mesenchymal transition of lens epithelium in mice Am J Physiol Cell Physiol, January 1, 2006; 290(1): C282 - C289. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Suzuki, Y. Daigo, N. Ishikawa, T. Kato, S. Hayama, T. Ito, E. Tsuchiya, and Y. Nakamura ANLN Plays a Critical Role in Human Lung Carcinogenesis through the Activation of RHOA and by Involvement in the Phosphoinositide 3-Kinase/AKT Pathway Cancer Res., December 15, 2005; 65(24): 11314 - 11325. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Zhang, M. Betson, J. Erasmus, K. Zeikos, M. Bailly, L. P. Cramer, and V. M. M. Braga Actin at cell-cell junctions is composed of two dynamic and functional populations J. Cell Sci., December 1, 2005; 118(23): 5549 - 5562. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. C. Wilkes, H. Mitchell, S. G. Penheiter, J. J. Dore, K. Suzuki, M. Edens, D. K. Sharma, R. E. Pagano, and E. B. Leof Transforming Growth Factor-{beta} Activation of Phosphatidylinositol 3-Kinase Is Independent of Smad2 and Smad3 and Regulates Fibroblast Responses via p21-Activated Kinase-2 Cancer Res., November 15, 2005; 65(22): 10431 - 10440. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Vogelmann, M.-D. Nguyen-tat, K. Giehl, G. Adler, D. Wedlich, and A. Menke TGF{beta}-induced downregulation of E-cadherin-based cell-cell adhesion depends on PI3-kinase and PTEN J. Cell Sci., October 15, 2005; 118(20): 4901 - 4912. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Levy and C. S. Hill Smad4 Dependency Defines Two Classes of Transforming Growth Factor {beta} (TGF-{beta}) Target Genes and Distinguishes TGF-{beta}-Induced Epithelial-Mesenchymal Transition from Its Antiproliferative and Migratory Responses Mol. Cell. Biol., September 15, 2005; 25(18): 8108 - 8125. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Deaton, C. Su, T. G. Valencia, and S. R. Grant Transforming Growth Factor-{beta}1-induced Expression of Smooth Muscle Marker Genes Involves Activation of PKN and p38 MAPK J. Biol. Chem., September 2, 2005; 280(35): 31172 - 31181. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T L Veveris-Lowe, M G Lawrence, R L Collard, L Bui, A C Herington, D L Nicol, and J A Clements Kallikrein 4 (hK4) and prostate-specific antigen (PSA) are associated with the loss of E-cadherin and an epithelial-mesenchymal transition (EMT)-like effect in prostate cancer cells Endocr. Relat. Cancer, September 1, 2005; 12(3): 631 - 643. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. N. Kloth, G. J. Fleuren, J. Oosting, R. X. de Menezes, P. H.C. Eilers, G. G. Kenter, and A. Gorter Substantial changes in gene expression of Wnt, MAPK and TNF{alpha} pathways induced by TGF-{beta}1 in cervical cancer cell lines Carcinogenesis, September 1, 2005; 26(9): 1493 - 1502. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Kang and K.K.H. Svoboda Epithelial-Mesenchymal Transformation during Craniofacial Development J. Dent. Res., August 1, 2005; 84(8): 678 - 690. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Yang, C. Zhong, B. Frenkel, A. H. Reddi, and P. Roy-Burman Diverse Biological Effect and Smad Signaling of Bone Morphogenetic Protein 7 in Prostate Tumor Cells Cancer Res., July 1, 2005; 65(13): 5769 - 5777. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Patel, K.-i. Takagi, J. Suzuki, A. Imaizumi, T. Kimura, R. M Mason, T. Kamimura, and Z. Zhang RhoGTPase Activation Is a Key Step in Renal Epithelial Mesenchymal Transdifferentiation J. Am. Soc. Nephrol., July 1, 2005; 16(7): 1977 - 1984. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
