 |
 Previous Article | Next Article 
Journal of Neuroscience, Vol 12, 86-95, Copyright © 1992 by Society for Neuroscience
Protons selectively induce lasting excitation and sensitization to mechanical stimulation of nociceptors in rat skin, in vitro
KH Steen, PW Reeh, F Anton and HO Handwerker
Institut fur Physiologie und Biokybernetik, Universitat Erlangen- Nurnberg, Germany.
In ischemic and in inflamed tissues, pH levels down to 5.4 have been
measured, and this local acidosis may contribute to pain and hyperalgesia
in disease states. To evaluate the role of acid pH in nociception, we have
studied identified primary afferents in a rat skin- saphenous nerve
preparation in vitro where the receptive fields can be superfused at the
highly permeable corium side with controlled solutions. The nerve endings
were exposed to CO2-saturated synthetic interstitial fluid (SIF;pH 6.1) and
to carbogen-gassed SIF phosphate buffered to different acid pH levels (5
min duration, 10 min intervals). Mechanical thresholds were repeatedly
tested in a "blind" fashion by von Frey hair stimulation. Low-threshold
mechanosensitive A beta- (n = 12) and A delta-fibers (n = 11) were not
excited or sensitized by acid pH levels. In 24 of 96 nociceptor type C- and
A delta-fibers, irregular low-frequency discharge with poor response
characteristics was induced. However, a distinct subpopulation of
mechanoheat sensitive, "polymodal" C-units (n = 25; 38%) showed
stimulus-related responses increasing with proton concentration and
encoding the time course of the pH change. Threshold levels were found to
range from pH 6.9 to 6.1; mean maximum discharge was at pH 5.2. All such
fibers responded to CO2 as well as to phosphate-buffered solution at the
same pH 6.1. The CO2 responses, however, displayed significantly shorter
latencies and more pronounced dynamic phases. The carboanhydrase blocker
acetazolamide markedly delayed and reduced the CO2 responses. Prolonged
application of acid pH (30 min) evoked nonadapting activity irrespective of
oxygen supply. Many, but certainly not all, fibers sensitive to protons
were also driven by capsaicin (10(- 6) M, 10(-5) M) and vice versa.
Repeated or prolonged treatment with low pH induced a significant and
lasting decrease of the mechanical (von Frey) thresholds in almost all
C-fibers tested (from 35 to 16 mN, on average), and this occurred whether
or not a fiber was excited by protons. The sensitizing effect was more
pronounced the higher the initial von Frey thresholds (0.75 rank
correlation). This sensitization to mechanical stimulation was in contrast
to the combined action of other inflammatory mediators, bradykinin, 5-HT,
histamine and prostaglandin E2. In conclusion, we suggest that pH
sensitivity of nociceptors may be an important source of pain and
hyperalgesia.
This article has been cited by other articles:
|
|
|
|
 
C. S. Surowy, T. R. Neelands, B. R. Bianchi, S. McGaraughty, R. El Kouhen, P. Han, K. L. Chu, H. A. McDonald, M. Vos, W. Niforatos, et al.
(R)-(5-tert-Butyl-2,3-dihydro-1H-inden-1-yl)-3-(1H-indazol-4-yl)-urea (ABT-102) Blocks Polymodal Activation of Transient Receptor Potential Vanilloid 1 Receptors in Vitro and Heat-Evoked Firing of Spinal Dorsal Horn Neurons in Vivo
J. Pharmacol. Exp. Ther.,
September 1, 2008;
326(3):
879 - 888.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. M. Cunha, W. A. Verri Jr., I. R. Schivo, M. H. Napimoga, C. A. Parada, S. Poole, M. M. Teixeira, S. H. Ferreira, and F. Q. Cunha
Crucial role of neutrophils in the development of mechanical inflammatory hypernociception
J. Leukoc. Biol.,
April 1, 2008;
83(4):
824 - 832.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Z. Gerevich, Z. S. Zadori, L. Koles, L. Kopp, D. Milius, K. Wirkner, K. Gyires, and P. Illes
Dual Effect of Acid pH on Purinergic P2X3 Receptors Depends on the Histidine 206 Residue
J. Biol. Chem.,
November 23, 2007;
282(47):
33949 - 33957.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. K. M. Lennerz, C. Dentsch, N. Bernardini, T. Hummel, W. L. Neuhuber, and P. W. Reeh
Electrophysiological characterization of vagal afferents relevant to mucosal nociception in the rat upper oesophagus
J. Physiol.,
July 1, 2007;
582(1):
229 - 242.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. Renner, G. Clarke, T. Grattan, A. Beisel, C. Mueller, U. Werner, G. Kobal, and K. Brune
Caffeine Accelerates Absorption and Enhances the Analgesic Effect of Acetaminophen
J. Clin. Pharmacol.,
June 1, 2007;
47(6):
715 - 726.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. E. Ferris, R. D. Clark, and E. L. Coates
Topical Inhibition of Nasal Carbonic Anhydrase Affects the CO2 Detection Threshold in Rats
Chem Senses,
March 1, 2007;
32(3):
263 - 271.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
O. Poirot, T. Berta, I. Decosterd, and S. Kellenberger
Distinct ASIC currents are expressed in rat putative nociceptors and are modulated by nerve injury
J. Physiol.,
October 1, 2006;
576(1):
215 - 234.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Jiang, K. K. Rau, R. D. Johnson, and B. Y. Cooper
Proton Sensitivity Ca2+ Permeability and Molecular Basis of Acid-Sensing Ion Channels Expressed in Glabrous and Hairy Skin Afferents
J Neurophysiol,
April 1, 2006;
95(4):
2466 - 2478.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Kim, C. Kang, C. Y. Shin, S. W. Hwang, Y. D. Yang, W. S. Shim, M.-Y. Park, E. Kim, M. Kim, B.-M. Kim, et al.
TRPV1 Recapitulates Native Capsaicin Receptor in Sensory Neurons in Association with Fas-Associated Factor 1
J. Neurosci.,
March 1, 2006;
26(9):
2403 - 2412.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Taguchi, J. Sato, and K. Mizumura
Augmented Mechanical Response of Muscle Thin-Fiber Sensory Receptors Recorded from Rat Muscle-Nerve Preparations In Vitro After Eccentric Contraction
J Neurophysiol,
October 1, 2005;
94(4):
2822 - 2831.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Jiang, M. Li, and L. Yue
Potentiation of TRPM7 Inward Currents by Protons
J. Gen. Physiol.,
July 25, 2005;
126(2):
137 - 150.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. K. Sauer, C. Weidner, R. W. Carr, B. Averbeck, U. Nesnidal, P. W. Reeh, and H. O. Handwerker
Can Receptor Potentials Be Detected With Threshold Tracking in Rat Cutaneous Nociceptive Terminals?
J Neurophysiol,
July 1, 2005;
94(1):
219 - 225.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Burstein and M. Jakubowski
Implications of multimechanism therapy: When to treat?
Neurology,
May 24, 2005;
64(10_suppl_2):
S16 - S20.
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Radhakrishnan and K. A. Sluka
Acetazolamide, a Carbonic Anhydrase Inhibitor, Reverses Inflammation-Induced Thermal Hyperalgesia in Rats
J. Pharmacol. Exp. Ther.,
May 1, 2005;
313(2):
921 - 927.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Dang, K. Bielefeldt, and G. F. Gebhart
Differential Responses of Bladder Lumbosacral and Thoracolumbar Dorsal Root Ganglion Neurons to Purinergic Agonists, Protons, and Capsaicin
J. Neurosci.,
April 13, 2005;
25(15):
3973 - 3984.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Schreckenberger, T. Siessmeier, A. Viertmann, C. Landvogt, H. -G. Buchholz, R. Rolke, R. -D. Treede, P. Bartenstein, and F. Birklein
The unpleasantness of tonic pain is encoded by the insular cortex
Neurology,
April 12, 2005;
64(7):
1175 - 1183.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Sugiura, K. Dang, K. Lamb, K. Bielefeldt, and G. F. Gebhart
Acid-Sensing Properties in Rat Gastric Sensory Neurons from Normal and Ulcerated Stomach
J. Neurosci.,
March 9, 2005;
25(10):
2617 - 2627.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Koulchitsky, M. J.M. Fischer, R. De Col, P. M. Schlechtweg, and K. Messlinger
Biphasic Response to Nitric Oxide of Spinal Trigeminal Neurons With Meningeal Input in Rat-Possible Implications for the Pathophysiology of Headaches
J Neurophysiol,
September 1, 2004;
92(3):
1320 - 1328.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Levy, M. Jakubowski, and R. Burstein
Disruption of communication between peripheral and central trigeminovascular neurons mediates the antimigraine action of 5HT1B/1D receptor agonists
PNAS,
March 23, 2004;
101(12):
4274 - 4279.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Ikoma, M. Fartasch, G. Heyer, Y. Miyachi, H. Handwerker, and M. Schmelz
Painful stimuli evoke itch in patients with chronic pruritus: Central sensitization for itch
Neurology,
January 27, 2004;
62(2):
212 - 217.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Mamet, M. Lazdunski, and N. Voilley
How Nerve Growth Factor Drives Physiological and Inflammatory Expressions of Acid-sensing Ion Channel 3 in Sensory Neurons
J. Biol. Chem.,
December 5, 2003;
278(49):
48907 - 48913.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. M. Wise, C. J. Wysocki, and T. Radil
Time-Intensity Ratings of Nasal Irritation from Carbon Dioxide
Chem Senses,
November 1, 2003;
28(9):
751 - 760.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. T. Hamamoto and D. A. Simone
Characterization of Cutaneous Primary Afferent Fibers Excited by Acetic Acid in a Model of Nociception in Frogs
J Neurophysiol,
August 1, 2003;
90(2):
566 - 577.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. J. Valenzano, E. R. Grant, G. Wu, M. Hachicha, L. Schmid, L. Tafesse, Q. Sun, Y. Rotshteyn, J. Francis, J. Limberis, et al.
N-(4-Tertiarybutylphenyl)-4-(3-chloropyridin-2-yl)tetrahydropyrazine -1(2H)-carbox-amide (BCTC), a Novel, Orally Effective Vanilloid Receptor 1 Antagonist with Analgesic Properties: I. In Vitro Characterization and Pharmacokinetic Properties
J. Pharmacol. Exp. Ther.,
July 1, 2003;
306(1):
377 - 386.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Distler, P. K. Rathee, K. S. Lips, O. Obreja, W. Neuhuber, and M. Kress
Fast Ca2+-Induced Potentiation of Heat-Activated Ionic Currents Requires cAMP/PKA Signaling and Functional AKAP Anchoring
J Neurophysiol,
May 1, 2003;
89(5):
2499 - 2505.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Orstavik, C. Weidner, R. Schmidt, M. Schmelz, M. Hilliges, E. Jorum, H. Handwerker, and E. Torebjork
Pathological C-fibres in patients with a chronic painful condition
Brain,
March 1, 2003;
126(3):
567 - 578.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Dirajlal, L. E. Pauers, and C. L. Stucky
Differential Response Properties of IB4-Positive and -Negative Unmyelinated Sensory Neurons to Protons and Capsaicin
J Neurophysiol,
January 1, 2003;
89(1):
513 - 524.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Mamet, A. Baron, M. Lazdunski, and N. Voilley
ProInflammatory Mediators, Stimulators of Sensory Neuron Excitability via the Expression of Acid-Sensing Ion Channels
J. Neurosci.,
December 15, 2002;
22(24):
10662 - 10670.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Gu and L.-Y. Lee
Alveolar hypercapnia augments pulmonary C-fiber responses to chemical stimulants: role of hydrogen ion
J Appl Physiol,
July 1, 2002;
93(1):
181 - 188.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C.-C. Chen, A. Zimmer, W.-H. Sun, J. Hall, M. J. Brownstein, and A. Zimmer
A role for ASIC3 in the modulation of high-intensity pain stimuli
PNAS,
June 25, 2002;
99(13):
8992 - 8997.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Xie, M. P. Price, A. L. Berger, and M. J. Welsh
DRASIC Contributes to pH-Gated Currents in Large Dorsal Root Ganglion Sensory Neurons by Forming Heteromultimeric Channels
J Neurophysiol,
June 1, 2002;
87(6):
2835 - 2843.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Koda and K. Mizumura
Sensitization to Mechanical Stimulation by Inflammatory Mediators and by Mild Burn in Canine Visceral Nociceptors In Vitro
J Neurophysiol,
April 1, 2002;
87(4):
2043 - 2051.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. M. Hamalainen, G. F. Gebhart, and T. J. Brennan
Acute Effect of an Incision on Mechanosensitive Afferents in the Plantar Rat Hindpaw
J Neurophysiol,
February 1, 2002;
87(2):
712 - 720.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Le Bars, M. Gozariu, and S. W. Cadden
Animal Models of Nociception
Pharmacol. Rev.,
December 1, 2001;
53(4):
597 - 652.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Malick, M. Jakubowski, J. K. Elmquist, C. B. Saper, and R. Burstein
A neurohistochemical blueprint for pain-induced loss of appetite
PNAS,
August 14, 2001;
98(17):
9930 - 9935.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. E. Cairns, J. W. Hu, L. Arendt-Nielsen, B. J. Sessle, and P. Svensson
Sex-Related Differences in Human Pain and Rat Afferent Discharge Evoked by Injection of Glutamate Into the Masseter Muscle
J Neurophysiol,
August 1, 2001;
86(2):
782 - 791.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-M. Zhang, Y. Homma, W. E. Ackerman, and S. J. Brull
Topical Application of Acidic Bupivacaine to the Lumbar Ganglion Induces Mechanical Hyperalgesia in the Rat
Anesth. Analg.,
August 1, 2001;
93(2):
466 - 471.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Garcia-Anoveros, T. A. Samad, L. Zuvela-Jelaska, C. J. Woolf, and D. P. Corey
Transport and Localization of the DEG/ENaC Ion Channel BNaC1{alpha} to Peripheral Mechanosensory Terminals of Dorsal Root Ganglia Neurons
J. Neurosci.,
April 15, 2001;
21(8):
2678 - 2686.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. A.S. Breslin, T. N. Gingrich, and B. G. Green
Ibuprofen as a Chemesthetic Stimulus: Evidence of a Novel Mechanism of Throat Irritation
Chem Senses,
January 1, 2001;
26(1):
55 - 65.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-M. Dessirier, M. O'Mahony, M. Iodi-Carstens, and E. Carstens
Sensory Properties of Citric Acid: Psychophysical Evidence for Sensitization, Self-desensitization, Cross-desensitization and Cross-stimulus-induced Recovery Following Capsaicin
Chem Senses,
December 1, 2000;
25(6):
769 - 780.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-M. Dessirier, C. T. Simons, M. I. Carstens, M. O'Mahony, and E. Carstens
Psychophysical and Neurobiological Evidence that the Oral Sensation Elicited by Carbonated Water is of Chemogenic Origin
Chem Senses,
June 1, 2000;
25(3):
277 - 284.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. J. Caterina, A. Leffler, A. B. Malmberg, W. J. Martin, J. Trafton, K. R. Petersen-Zeitz, M. Koltzenburg, A. I. Basbaum, and D. Julius
Impaired Nociception and Pain Sensation in Mice Lacking the Capsaicin Receptor
Science,
April 14, 2000;
288(5464):
306 - 313.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
H. Hirata, J. W. Hu, and D. A. Bereiter
Responses of Medullary Dorsal Horn Neurons to Corneal Stimulation by CO2 Pulses in the Rat
J Neurophysiol,
November 1, 1999;
82(5):
2092 - 2107.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. T. Simons, J. -M. Dessirier, M. I. Carstens, M. O'Mahony, and E. Carstens
Neurobiological and Psychophysical Mechanisms Underlying the Oral Sensation Produced by Carbonated Water
J. Neurosci.,
September 15, 1999;
19(18):
8134 - 8144.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Honore, P. M. Menning, S. D. Rogers, M. L. Nichols, A. I. Basbaum, J.-M. Besson, and P. W. Mantyh
Spinal Substance P Receptor Expression and Internalization in Acute, Short-Term, and Long-Term Inflammatory Pain States
J. Neurosci.,
September 1, 1999;
19(17):
7670 - 7678.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. J. Mannion, M. Costigan, I. Decosterd, F. Amaya, Q.-P. Ma, J. C. Holstege, R.-R. Ji, A. Acheson, R. M. Lindsay, G. A. Wilkinson, et al.
Neurotrophins: Peripherally and centrally acting modulators of tactile stimulus-induced inflammatory pain hypersensitivity
PNAS,
August 3, 1999;
96(16):
9385 - 9390.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. L. Durham and A. F. Russo
Regulation of Calcitonin Gene-Related Peptide Secretion by a Serotonergic Antimigraine Drug
J. Neurosci.,
May 1, 1999;
19(9):
3423 - 3429.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Yamamura, A. Malick, N. L. Chamberlin, and R. Burstein
Cardiovascular and Neuronal Responses to Head Stimulation Reflect Central Sensitization and Cutaneous Allodynia in a Rat Model of Migraine
J Neurophysiol,
February 1, 1999;
81(2):
479 - 493.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Jung, S. W. Hwang, J. Kwak, S.-Y. Lee, C.-J. Kang, W. B. Kim, D. Kim, and U. Oh
Capsaicin Binds to the Intracellular Domain of the Capsaicin-Activated Ion Channel
J. Neurosci.,
January 15, 1999;
19(2):
529 - 538.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C.-C. Chen, S. England, A. N. Akopian, and J. N. Wood
A sensory neuron-specific, proton-gated ion channel
PNAS,
August 18, 1998;
95(17):
10240 - 10245.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Carstens, N. Kuenzler, and H. O. Handwerker
Activation of Neurons in Rat Trigeminal Subnucleus Caudalis by Different Irritant Chemicals Applied to Oral or Ocular Mucosa
J Neurophysiol,
August 1, 1998;
80(2):
465 - 492.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. DAOUI, C. COGNON, E. NALINE, X. EMONDS-ALT, and C. ADVENIER
Involvement of Tachykinin NK3 Receptors in Citric Acid-induced Cough and Bronchial Responses in Guinea Pigs
Am. J. Respir. Crit. Care Med.,
July 1, 1998;
158(1):
42 - 48.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Vyklicky, H. Knotkova-Urbancova, Z. Vitaskova, V. Vlachova, M. Kress, and P. W. Reeh
Inflammatory Mediators at Acidic pH Activate Capsaicin Receptors in Cultured Sensory Neurons From Newborn Rats
J Neurophysiol,
February 1, 1998;
79(2):
670 - 676.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Burstein, H. Yamamura, A. Malick, and A. M. Strassman
Chemical Stimulation of the Intracranial Dura Induces Enhanced Responses to Facial Stimulation in Brain Stem Trigeminal Neurons
J Neurophysiol,
February 1, 1998;
79(2):
964 - 982.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Waldmann, F. Bassilana, J. de Weille, G. Champigny, C. Heurteaux, and M. Lazdunski
Molecular Cloning of a Non-inactivating Proton-gated Na+ Channel Specific for Sensory Neurons
J. Biol. Chem.,
August 22, 1997;
272(34):
20975 - 20978.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Ebersberger, M. Ringkamp, P. W. Reeh, and H. O. Handwerker
Recordings From Brain Stem Neurons Responding to Chemical Stimulation of the Subarachnoid Space
J Neurophysiol,
June 1, 1997;
77(6):
3122 - 3133.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. M. Bove and M. A. Moskowitz
Primary Afferent Neurons Innervating Guinea Pig Dura
J Neurophysiol,
January 1, 1997;
77(1):
299 - 308.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Kress and P. W. Reeh
More sensory competence for nociceptive neurons in culture
PNAS,
December 24, 1996;
93(26):
14995 - 14997.
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. J. Welsh, M. P. Price, and J. Xie
Biochemical Basis of Touch Perception: Mechanosensory Function of Degenerin/Epithelial Na+ Channels
J. Biol. Chem.,
January 18, 2002;
277(4):
2369 - 2372.
[Full Text]
[PDF]
|
|
|
|
|
|
| |
