Circulation. 2003;107:1017-1023
Published online before print February 3, 2003, doi: 10.1161/01.CIR.0000051367.92927.07
Published online before print February 3, 2003, doi: 10.1161/01.CIR.0000051367.92927.07
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
(Circulation. 2003;107:1017.)
© 2003 American Heart Association, Inc.
Basic Science Reports |
High Glucose Causes Upregulation of Cyclooxygenase-2 and Alters Prostanoid Profile in Human Endothelial Cells
Role of Protein Kinase C and Reactive Oxygen Species
From the Department of Cardiovascular Research, Institute of Physiology (F.C., M.E., B.v.d.L., A.K., C.D.G., H.J., T.F.L.), University of Zurich and Cardiovascular Center, University Hospital, Zurich, Switzerland; Department of Experimental Medicine and Pathology (F.C., P.D.P., M.V.), University "La Sapienza," Rome and IRCCS Neuromed, Pozzilli, Italy; and Faculty of Biology (M.B., V.U.), University of Konstanz, Germany.
Correspondence to Francesco Cosentino, MD, PhD, Cardiovascular Research, Institute of Physiology, University of Zurich-Irchel, CH-8057 Zürich, Switzerland. E-mail f_cosentino{at}hotmail.com
Background— Prostaglandins generated by cyclooxygenase (COX) have been implicated in hyperglycemia-induced endothelial dysfunction. However, the role of individual COX isoenzymes as well as the molecular mechanisms linking oxidative stress and endothelial dysfunction in diabetes remains to be clarified.
Methods and Results— Human aortic endothelial cells were exposed to normal (5.5 mmol/L) and high (22.2 mmol/L) glucose. Glucose selectively increased mRNA and protein expression of COX-2. Its upregulation was associated with an increase of thromboxane A2 and a reduction of prostacyclin (PGI2) release. Glucose-induced activation of PKC resulted in the formation of peroxynitrite and tyrosine nitration of PGI2 synthase. NO release was reduced despite 2-fold increase of endothelial NO synthase expression. Phorbol ester caused an increase of COX-2 and endothelial NO synthase expression similar to that elicited by glucose. These effects were prevented by the PKC inhibitor calphostin C. N-acetylcysteine, vitamin C, and calphostin C prevented ROS formation, restored NO release, and reduced colocalization of nitrotyrosine and PGI2 synthase. Expression of p22phox, a subunit of NAD(P)H oxidase, was increased, and diphenyleneiodonium inhibited ROS formation. By contrast, indomethacin did not affect glucose-induced ROS generation.
Conclusions— Thus, high glucose, via PKC signaling, induces oxidative stress and upregulation of COX-2, resulting in reduced NO availability and altered prostanoid profile.
Key Words: diabetes mellitus • nitric oxide • stress
This article has been cited by other articles:
 |
|
 |
|
  J.-L. Balligand, O. Feron, and C. Dessy eNOS Activation by Physical Forces: From Short-Term Regulation of Contraction to Chronic Remodeling of Cardiovascular Tissues Physiol Rev, April 1, 2009; 89(2): 481 - 534. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Labinskyy, P. Mukhopadhyay, J. Toth, G. Szalai, M. Veres, G. Losonczy, J. T. Pinto, P. Pacher, P. Ballabh, A. Podlutsky, et al. Longevity is associated with increased vascular resistance to high glucose-induced oxidative stress and inflammatory gene expression in Peromyscus leucopus Am J Physiol Heart Circ Physiol, April 1, 2009; 296(4): H946 - H956. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. L. Hodnett, J. A. Dearman, C. B. Carter, and R. L. Hester Attenuated PGI2 synthesis in obese Zucker rats Am J Physiol Regulatory Integrative Comp Physiol, March 1, 2009; 296(3): R715 - R721. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Nacci, M. Tarquinio, L. De Benedictis, A. Mauro, A. Zigrino, M. R. Carratu, M. J. Quon, and M. Montagnani Endothelial Dysfunction in Mice with Streptozotocin-induced Type 1 Diabetes Is Opposed by Compensatory Overexpression of Cyclooxygenase-2 in the Vasculature Endocrinology, February 1, 2009; 150(2): 849 - 861. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. P. Kellogg, K. Converso, T. Wiggin, M. Stevens, and R. Pop-Busui Effects of cyclooxygenase-2 gene inactivation on cardiac autonomic and left ventricular function in experimental diabetes Am J Physiol Heart Circ Physiol, February 1, 2009; 296(2): H453 - H461. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Cosentino, R. Battista, A. Scuteri, F. De Sensi, L. De Siati, C. Di Russo, G. G. Camici, and M. Volpe Impact of Fasting Glycemia and Regional Cerebral Perfusion in Diabetic Subjects: A Study With Technetium-99m-Ethyl Cysteinate Dimer Single Photon Emission Computed Tomography Stroke, January 1, 2009; 40(1): 306 - 308. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Shanmugam, M. A. Reddy, and R. Natarajan Distinct Roles of Heterogeneous Nuclear Ribonuclear Protein K and microRNA-16 in Cyclooxygenase-2 RNA Stability Induced by S100b, a Ligand of the Receptor for Advanced Glycation End Products J. Biol. Chem., December 26, 2008; 283(52): 36221 - 36233. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. S. Wilcox and A. Pearlman Chemistry and Antihypertensive Effects of Tempol and Other Nitroxides Pharmacol. Rev., December 1, 2008; 60(4): 418 - 469. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Osto, A. Kouroedov, P. Mocharla, A. Akhmedov, C. Besler, L. Rohrer, A. von Eckardstein, S. Iliceto, M. Volpe, T. F. Luscher, et al. Inhibition of Protein Kinase C{beta} Prevents Foam Cell Formation by Reducing Scavenger Receptor A Expression in Human Macrophages Circulation, November 18, 2008; 118(21): 2174 - 2182. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Li, L. Kuo, and J. N. Stallone Estrogen potentiates constrictor prostanoid function in female rat aorta by upregulation of cyclooxygenase-2 and thromboxane pathway expression Am J Physiol Heart Circ Physiol, June 1, 2008; 294(6): H2444 - H2455. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Syeda, E. Tullis, A. S. Slutsky, and H. Zhang Human neutrophil peptides upregulate expression of COX-2 and endothelin-1 by inducing oxidative stress Am J Physiol Heart Circ Physiol, June 1, 2008; 294(6): H2769 - H2774. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Cosentino, P. Francia, G. G. Camici, P. G. Pelicci, M. Volpe, and T. F. Luscher Final Common Molecular Pathways of Aging and Cardiovascular Disease: Role of the p66Shc Protein Arterioscler. Thromb. Vasc. Biol., April 1, 2008; 28(4): 622 - 628. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Xiang, J. Dearman, S. R. Abram, C. Carter, and R. L. Hester Insulin resistance and impaired functional vasodilation in obese Zucker rats Am J Physiol Heart Circ Physiol, April 1, 2008; 294(4): H1658 - H1666. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y.-J. Chen and J. Quilley Fenofibrate Treatment of Diabetic Rats Reduces Nitrosative Stress, Renal Cyclooxygenase-2 Expression, and Enhanced Renal Prostaglandin Release J. Pharmacol. Exp. Ther., February 1, 2008; 324(2): 658 - 663. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. P. Kellogg, T. D. Wiggin, D. D. Larkin, J. M. Hayes, M. J. Stevens, and R. Pop-Busui Protective Effects of Cyclooxygenase-2 Gene Inactivation Against Peripheral Nerve Dysfunction and Intraepidermal Nerve Fiber Loss in Experimental Diabetes Diabetes, December 1, 2007; 56(12): 2997 - 3005. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Quijano, L. Castro, G. Peluffo, V. Valez, and R. Radi Enhanced mitochondrial superoxide in hyperglycemic endothelial cells: direct measurements and formation of hydrogen peroxide and peroxynitrite Am J Physiol Heart Circ Physiol, December 1, 2007; 293(6): H3404 - H3414. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Authors/Task Force Members, L. Ryden, E. Standl, M. Bartnik, G. V. d. Berghe, J. Betteridge, M.-J. de Boer, F. Cosentino, B. Jonsson, M. Laakso, et al. Guidelines on diabetes, pre-diabetes, and cardiovascular diseases: full text: The Task Force on Diabetes and Cardiovascular Diseases of the European Society of Cardiology (ESC) and of the European Association for the Study of Diabetes (EASD) Eur. Heart J. Suppl., June 1, 2007; 9(suppl_C): C3 - C74. [Full Text] [PDF]
|
|
|
|
|
|
Y. S. Park, J. Kim, Y. Misonou, R. Takamiya, M. Takahashi, M. R. Freeman, and N. Taniguchi Acrolein Induces Cyclooxygenase-2 and Prostaglandin Production in Human Umbilical Vein Endothelial Cells: Roles of p38 MAP Kinase Arterioscler. Thromb. Vasc. Biol., June 1, 2007; 27(6): 1319 - 1325. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-H. Chen, S.-J. Lin, F.-Y. Lin, T.-C. Wu, C.-R. Tsao, P.-H. Huang, P.-L. Liu, Y.-L. Chen, and J.-W. Chen High Glucose Impairs Early and Late Endothelial Progenitor Cells by Modifying Nitric Oxide-Related but Not Oxidative Stress-Mediated Mechanisms Diabetes, June 1, 2007; 56(6): 1559 - 1568. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. You, A. Ren, G. Yan, and J. Sun Activation of Sphingosine Kinase-1 Mediates Inhibition of Vascular Smooth Muscle Cell Apoptosis by Hyperglycemia Diabetes, May 1, 2007; 56(5): 1445 - 1453. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Guo, Z. Xia, J. Jiang, and J. H. McNeill Downregulation of NADPH oxidase, antioxidant enzymes, and inflammatory markers in the heart of streptozotocin-induced diabetic rats by N-acetyl-L-cysteine Am J Physiol Heart Circ Physiol, April 1, 2007; 292(4): H1728 - H1736. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. E. Kanter, F. Johansson, R. C. LeBoeuf, and K. E. Bornfeldt Do Glucose and Lipids Exert Independent Effects on Atherosclerotic Lesion Initiation or Progression to Advanced Plaques? Circ. Res., March 30, 2007; 100(6): 769 - 781. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. G. Camici, M. Schiavoni, P. Francia, M. Bachschmid, I. Martin-Padura, M. Hersberger, F. C. Tanner, P. Pelicci, M. Volpe, P. Anversa, et al. Genetic deletion of p66Shc adaptor protein prevents hyperglycemia-induced endothelial dysfunction and oxidative stress PNAS, March 20, 2007; 104(12): 5217 - 5222. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Szerafin, N. Erdei, T. Fulop, E. T. Pasztor, I. Edes, A. Koller, and Z. Bagi Increased Cyclooxygenase-2 Expression and Prostaglandin-Mediated Dilation in Coronary Arterioles of Patients With Diabetes Mellitus Circ. Res., September 1, 2006; 99(5): e12 - 317. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. W. Smith, R. L. Gamelli, S. B. Jones, and R. Shankar Immunologic Responses to Critical Injury and Sepsis J Intensive Care Med, May 1, 2006; 21(3): 160 - 172. [Abstract] [PDF]
|
|
|
|
|
|
C. Vecchione, A. Aretini, G. Marino, U. Bettarini, R. Poulet, A. Maffei, M. Sbroggio, L. Pastore, M. T. Gentile, A. Notte, et al. Selective Rac-1 Inhibition Protects From Diabetes-Induced Vascular Injury Circ. Res., February 3, 2006; 98(2): 218 - 225. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Zuccollo, C. Shi, R. Mastroianni, K. A. Maitland-Toolan, R. M. Weisbrod, M. Zang, S. Xu, B. Jiang, J. M. Oliver-Krasinski, A. J. Cayatte, et al. The Thromboxane A2 Receptor Antagonist S18886 Prevents Enhanced Atherogenesis Caused by Diabetes Mellitus Circulation, November 8, 2005; 112(19): 3001 - 3008. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Guo, W. Su, S. Allen, H. Pang, A. Daugherty, E. Smart, and M. C. Gong COX-2 Up-regulation and vascular smooth muscle contractile hyperreactivity in spontaneous diabetic db/db mice Cardiovasc Res, September 1, 2005; 67(4): 723 - 735. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. A. Berkowitz, R. Roberts, H. Luan, J. Peysakhov, D. L. Knoerzer, J. R. Connor, and T. C. Hohman Drug Intervention Can Correct Subnormal Retinal Oxygenation Response in Experimental Diabetic Retinopathy Invest. Ophthalmol. Vis. Sci., August 1, 2005; 46(8): 2954 - 2960. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Li, Y.-J. Chen, and J. Quilley Effect of Tempol on Renal Cyclooxygenase Expression and Activity in Experimental Diabetes in the Rat J. Pharmacol. Exp. Ther., August 1, 2005; 314(2): 818 - 824. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Bagi, N. Erdei, A. Toth, W. Li, T. H. Hintze, A. Koller, and G. Kaley Type 2 Diabetic Mice Have Increased Arteriolar Tone and Blood Pressure: Enhanced Release of COX-2-Derived Constrictor Prostaglandins Arterioscler. Thromb. Vasc. Biol., August 1, 2005; 25(8): 1610 - 1616. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Capaldo, M. Galderisi, A. A. Turco, A. D'Errico, S. Turco, A. A. Rivellese, G. de Simone, O. de Divitiis, and G. Riccardi Acute Hyperglycemia Does Not Affect the Reactivity of Coronary Microcirculation in Humans J. Clin. Endocrinol. Metab., July 1, 2005; 90(7): 3871 - 3876. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Lu, X.-L. Wang, T. He, W. Zhou, T. L. Kaduce, Z. S. Katusic, A. A. Spector, and H.-C. Lee Impaired Arachidonic Acid-Mediated Activation of Large-Conductance Ca2+-Activated K+ Channels in Coronary Arterial Smooth Muscle Cells in Zucker Diabetic Fatty Rats Diabetes, July 1, 2005; 54(7): 2155 - 2163. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Rikitake and J. K. Liao Rho-Kinase Mediates Hyperglycemia-Induced Plasminogen Activator Inhibitor-1 Expression in Vascular Endothelial Cells Circulation, June 21, 2005; 111(24): 3261 - 3268. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. S. O'Neill, A. Veves, A. Zanobetti, J. A. Sarnat, D. R. Gold, P. A. Economides, E. S. Horton, and J. Schwartz Diabetes Enhances Vulnerability to Particulate Air Pollution-Associated Impairment in Vascular Reactivity and Endothelial Function Circulation, June 7, 2005; 111(22): 2913 - 2920. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. L. Moens, I. Goovaerts, M. J. Claeys, and C. J. Vrints Flow-Mediated Vasodilation: A Diagnostic Instrument, or an Experimental Tool? Chest, June 1, 2005; 127(6): 2254 - 2263. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Zhou, X.-L. Wang, T. L. Kaduce, A. A. Spector, and H.-C. Lee Impaired arachidonic acid-mediated dilation of small mesenteric arteries in Zucker diabetic fatty rats Am J Physiol Heart Circ Physiol, May 1, 2005; 288(5): H2210 - H2218. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Xiang, J. Naik, and R. L. Hester Exercise-induced increase in skeletal muscle vasodilatory responses in obese Zucker rats Am J Physiol Regulatory Integrative Comp Physiol, April 1, 2005; 288(4): R987 - R991. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Rask-Madsen and G. L. King Proatherosclerotic Mechanisms Involving Protein Kinase C in Diabetes and Insulin Resistance Arterioscler. Thromb. Vasc. Biol., March 1, 2005; 25(3): 487 - 496. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. L. Sheu, F. M. Ho, R. S. Yang, K. F. Chao, W. W. Lin, S. Y. Lin-Shiau, and S.-H. Liu High Glucose Induces Human Endothelial Cell Apoptosis Through a Phosphoinositide 3-Kinase-Regulated Cyclooxygenase-2 Pathway Arterioscler. Thromb. Vasc. Biol., March 1, 2005; 25(3): 539 - 545. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Cai, J. Khoo, and K. M. Channon Augmented BH4 by gene transfer restores nitric oxide synthase function in hyperglycemic human endothelial cells Cardiovasc Res, March 1, 2005; 65(4): 823 - 831. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Beloqui, J. A. Paramo, J. Orbe, A. Benito, I. Colina, A. Monasterio, and J. Diez Monocyte cyclooxygenase-2 overactivity: a new marker of subclinical atherosclerosis in asymptomatic subjects with cardiovascular risk factors? Eur. Heart J., January 2, 2005; 26(2): 153 - 158. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. R. Heitmeier, C. B. Kelly, N. J. Ensor, K. A. Gibson, K. G. Mullis, J. A. Corbett, and T. J. Maziasz Role of Cyclooxygenase-2 in Cytokine-induced {beta}-Cell Dysfunction and Damage by Isolated Rat and Human Islets J. Biol. Chem., December 17, 2004; 279(51): 53145 - 53151. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Kinoshita, T. Azma, K. Nakahata, H. Iranami, Y. Kimoto, M. Dojo, O. Yuge, and Y. Hatano Inhibitory Effect of High Concentration of Glucose on Relaxations to Activation of ATP-Sensitive K+ Channels in Human Omental Artery Arterioscler. Thromb. Vasc. Biol., December 1, 2004; 24(12): 2290 - 2295. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. N. Bratz and N. L. Kanagy Nitric oxide synthase-inhibition hypertension is associated with altered endothelial cyclooxygenase function Am J Physiol Heart Circ Physiol, December 1, 2004; 287(6): H2394 - H2401. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-D. Luo, Y.-Y. Wang, W.-L. Fu, J. Wu, and A. F. Chen Gene Therapy of Endothelial Nitric Oxide Synthase and Manganese Superoxide Dismutase Restores Delayed Wound Healing in Type 1 Diabetic Mice Circulation, October 19, 2004; 110(16): 2484 - 2493. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Nasrallah and R. L. Hebert Reduced IP receptors in STZ-induced diabetic rat kidneys and high-glucose-treated mesangial cells Am J Physiol Renal Physiol, October 1, 2004; 287(4): F673 - F681. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Natarajan and J. L. Nadler Lipid Inflammatory Mediators in Diabetic Vascular Disease Arterioscler. Thromb. Vasc. Biol., September 1, 2004; 24(9): 1542 - 1548. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. He and G. L. King Protein Kinase C{beta} Isoform Inhibitors: A New Treatment for Diabetic Cardiovascular Diseases Circulation, July 6, 2004; 110(1): 7 - 9. [Full Text] [PDF]
|
|
|
|
|
|
A. Kouroedov, M. Eto, H. Joch, M. Volpe, T. F. Luscher, and F. Cosentino Selective Inhibition of Protein Kinase C{beta}2 Prevents Acute Effects of High Glucose on Vascular Cell Adhesion Molecule-1 Expression in Human Endothelial Cells Circulation, July 6, 2004; 110(1): 91 - 96. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Kusaka, G. Kusaka, C. Zhou, M. Ishikawa, A. Nanda, D. N. Granger, J. H. Zhang, and J. Tang Role of AT1 receptors and NAD(P)H oxidase in diabetes-aggravated ischemic brain injury Am J Physiol Heart Circ Physiol, June 1, 2004; 286(6): H2442 - H2451. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Radi Nitric oxide, oxidants, and protein tyrosine nitration PNAS, March 23, 2004; 101(12): 4003 - 4008. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Shanmugam, I. T. Gaw Gonzalo, and R. Natarajan Molecular Mechanisms of High Glucose-Induced Cyclooxygenase-2 Expression in Monocytes Diabetes, March 1, 2004; 53(3): 795 - 802. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Raitakari, T. Ilvonen, M. Ahotupa, T. Lehtimaki, A. Harmoinen, P. Suominen, J. Elo, J. Hartiala, and O. T. Raitakari Weight Reduction With Very-Low-Caloric Diet and Endothelial Function in Overweight Adults: Role of Plasma Glucose Arterioscler. Thromb. Vasc. Biol., January 1, 2004; 24(1): 124 - 128. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Creager, T. F. Luscher, F. Cosentino, and J. A. Beckman Diabetes and Vascular Disease: Pathophysiology, Clinical Consequences, and Medical Therapy: Part I Circulation, September 23, 2003; 108(12): 1527 - 1532. [Full Text] [PDF]
|
|
|
|
|
|
N. Shanmugam, Y. S. Kim, L. Lanting, and R. Natarajan Regulation of Cyclooxygenase-2 Expression in Monocytes by Ligation of the Receptor for Advanced Glycation End Products J. Biol. Chem., September 12, 2003; 278(37): 34834 - 34844. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Marfella, K. Esposito, D. Giugliano, F. Cosentino, M. Eto, B. van der Loo, A. Kouroedov, C. Delli Gatti, H. Joch, T. F. Luscher, et al. Effect of High Glucose on Vasculature * Response Circulation, September 9, 2003; 108 (10): e74 - e74. [Full Text] [PDF]
|
|