進階搜尋


 
系統識別號 U0026-0812200911164445
論文名稱(中文) 利用Fos免疫染色探討水楊酸誘發耳鳴在腦幹之作用位置
論文名稱(英文) Origins of salicylate-induced tinnitus in the auditory brainstem of rats revealed by Fos immunohistochemistry
校院名稱 成功大學
系所名稱(中) 生理學研究所
系所名稱(英) Department of Physiology
學年度 92
學期 2
出版年 93
研究生(中文) 朱育靜
研究生(英文) Yu-Ching Chu
電子信箱 jefy6823@yahoo.com.tw
學號 s3690101
學位類別 碩士
語文別 英文
論文頁數 44頁
口試委員 召集委員-華 瑜
指導教授-潘偉豐
口試委員-廖寶琦
中文關鍵字 耳鳴  電灑游離串聯式質譜儀  側上橄欖體神經核  下丘  水楊酸 
英文關鍵字 Fos  lateral superior olivary nucleus  ESI-MS/MS  salicylate  tinnitus  inferior colliculus 
學科別分類
中文摘要   耳鳴是一種在無外在音源下所產生的聽覺,目前對於其生理病理成因尚未清楚。在人類及動物以證實在過量水楊酸下會造成可逆性耳鳴。在中耳局部投與水楊酸實驗中已發現由對聲音反應的聽腦幹誘發電位受到抑制亦支持水楊酸在耳蝸之作用。全身性投與水楊酸則造成中樞聽神經元的過度活化。對於此周邊及中樞作用之矛盾現象目前仍未有清楚的解釋。本研究欲在探討經由兩種不同途徑長期持續投與水楊酸對中樞聽覺核活性所造成的反應,藉由即時表現基因c-fos 之蛋白質產物Fos 為神經元活性標記,利用Fos免疫染色法呈現出聽腦幹核之活性改變。
  本研究實驗大鼠(Sprague Dawley rat)分成兩組分別進行無菌手術。腹腔持續釋藥系統組以將注滿不同濃度(250及360 mg/kg/day)的水楊酸溶液之滲透幫浦(osmotic pump)置入腹腔內7 天;中耳持續釋藥系統組則以吸收不同濃度(5, 50,及250 mM)的水楊酸膠海綿放入中耳腔內6天,並分別以溶媒組實驗動物為對照組。實驗動物最後取腦做冷凍切片及Fos免疫染色並採用ABC呈色法。經顯微鏡觀察聽腦幹區耳蝸核(cochlear nucleus)、下丘(inferior colliculus)、及側上橄欖核(lateral superior olivary nucleus)區之Fos免疫染色分布及量化結果來觀察神經活性變化。另外為得知水楊酸局部(外耳淋巴液)及中樞(腦脊液)濃度對Fos表現之影響以及不同時間點之濃度,本研究另利用電灑游離串聯式質譜分析方法進一步來分析水楊酸濃度。
  實驗結果顯示經由中耳及腹腔持續釋藥這兩種給藥方式均能觀察到Fos表現在下丘及側上橄欖核均有似連續5天單一劑量水楊酸注射的明顯增加現象,此亦證實耳鳴發生的可能性。中耳釋藥組中外耳淋巴液水楊酸含量將快速上升而隨著時間腦脊液中亦有緩升的水楊酸濃度,此證實在被動性通透的存在,例如耳蝸導水管。腹腔投與組中外耳淋巴液水楊酸含量亦快速上升且高於腦脊液,其可能因耳蝸有著主動分泌機制。因此周邊耳蝸水楊酸投與,即使是單側,可引起之耳鳴現象,其可支持周邊作用可能亦包括著但需中樞作用的存在。此活化現象主要是作傳導閥值的微調。對照組動物的Fos相對高數量可能來自於寂靜環境處理下之生理性耳鳴,其可能於多日後因適應而消失。在耳蝸核中,所有實驗動物Fos的表現並不會增加,此亦意味著耳鳴的來源主要來自於中樞而非周邊傳入。因此本研究亦證實了水楊酸引起之耳鳴成因是先作用於外周耳蝸進而影響中樞聽覺系統之過度活化所致。


英文摘要   Tinnitus is a phantom sensation of sound in the absence of external stimulations and its pathogenesis is unclear. Overdose of salicylate (SA) is known to cause tinnitus in both human and animals. Local application of SA in the bulla suppresses evoked potential to sound suggesting a peripheral action of SA at the cochlea. Systemic application of SA also induces over-activities of central auditory neurons suggesting a central action. In this study, the effects of SA applied at two different routes were compared in order to elucidate the action sites. Fos-immunohistochemistry was used to reveal the hyper-activity in the brainstem nuclei. Adult rats were divided into two groups for aseptic surgery. The first group was intraperitoneally implanted with osmotic pumps that would sustain release SA for 7 days. The second group had a SA-presoaked gelfoam placed over the round window in the bullae for 6 days. Controls received similar procedures with vehicle-treatments. Brains were processed for Fos-immunohistochemistry on thick frozen sections according to the conventional ABC method. For comparison, a limited number of animals also received a 5-daily SA injection. For some animals, SA levels in the perilymph and cerebrospinal fluid were measured at different time points with mass spectrometry (ESI-MS/MS).
  Both the osmotic pump and gelfoam applications that released SA slowly were effective in raising Fos-activity labeling both in the inferior colliculus and lateral superior olivary complex in a manner rather similar to the multiple-daily injection protocol, suggesting a possible occurrence of tinnitus. Following intra-bullar applications there was a rapid increase of SA in perilymph followed by a delayed increase in CSF to about the level, suggesting an effective fluid passage linking the two compartments likely through the cochlear aqueduct. Intra-peritoneal applications also led to a rapid rise in SA level in the perilymph at a level higher than the CSF, suggesting the presence of an active secretion mechanism in the cochlea. The fact that SA when applied locally to the cochlea, even on one side, was effective suggested that a peripheral site was likely involved but may also require the action at the central nervous system. The relatively high level of Fos-expression at the IC of control rats was discussed with respect to a physiological tinnitus, which might subside in Fos-expression after animals were adapted to a silent environment.


論文目次 Abstract
Abstract (in Chinese)

1. Introduction
1.1 Tinnitus 1
1.2 What is the relationship between salicylate
(SA) and tinnitus in human? 1
1.3 Animal studies on SA-induced tinnitus 2
1.4 The auditory brainstem and midbrain versus
SA-induced tinnitus 3
1.4.1 Inferior colliculus (IC) 3
1.4.2 Lateral olivary nucleus (LSO) and cochlea
nucleus (CN) 3
1.4.3 SA & auditory nuclei 3
1.5 Fos as a neural activity marker 4
1.6 Methods of SA measurement 5
1.7 Aims of this study 6

2. Materials
2.1 Instruments for online clean-up device with
ESI-MS/MS system 8
2.2 Chemicals and materials 9
2.3 Immunohistochemistry 10
2.4 Anesthetics 10
2.5 Solutions preparation 11

3. Methods
3.1 Establishing the SA analytical method 15
3.1.1 Online clean-up device and ESI-MS/MS system
15
3.1.2 Optimization of MS instrument 15
3.1.3 Online clean-up system 17
3.1.4 Validation of the analytic method 17
3.1.5 Analysis of the biological samples 20
3.1.6 Pre-treatment of the bio-sample 20
3.2 Animal preparation 21
3.2.1 Acute experiments 21
3.2.2 Chronic experiments 22
3.3 Perfusion and tissue preparation for Fos
Immunohistochemistry 23
3.4 Fos Immunohistochemistry 23
3.5 Data analysis 24
3.5.1 The counts and location of Fos-labeled
neurons 24
3.5.2 3D reconstruction of the Fos-labeled
neurons 24

4. Results
4.1 Establishing the SA analytical method 26
4.1.1 Validation of the analytic method 26
4.1.1.1 Selectivity 26
4.1.1.2 MDL (method detection limit) 26
4.1.1.3 Calibration and Linearity 26
4.1.1.4 Precision 27
4.1.1.5 Recovery 27
4.2 Time profiles of SA levels in perilymph and
cerebral spinal fluid (CSF) after acute
applications of SA 28
4.2.1 Intra-bulla application with SA-presoaked
gelfoam 28
4.2.2 Peritoneal application with SA single dose
injection 28
4.3 Residual SA levels in perilymph and CSF
after chronic application of SA via
intra-bulla or intra-peritoneal routes
28
4.4 Side effects of chronic SA application 29
4.5 Fos-activity changes in the inferior
colliculus (IC) 30
4.5.1 Chronic intra-bulla application of SA 30
4.5.2 Chronic intra-peritoneal application of SA
30
4.6 Fos-activity changes in the cochlear
nucleus (CN) 30
4.7 Fos-activity changes in the lateral
superior olive (LSO) 31

5. Discussion
5.1. The validation results of the analytic
method for SA 32
5.1.1. Recovery 32
5.1.2. Precision 32
5.2. The profile of SA in perilymph and CSF
after SA acute application 32
5.3. SA residual in perilymph and CSF at the end
of SA chronic applications 33
5.4. SA dose is enough to induce tinnitus-like
pattern 33
5.5. Side effects of chronic SA application 33
5.6. The possible mechanism of salicylate-
induce tinnitus-like pattern in the IC 34
5.7. Fos expression in the high-frequency region
at the CIC 34
5.8. Tinnitus-like Fos pattern in the IC of
control rats 35

6. Summary 37
7. References 38
8. Tables 44




參考文獻 Aitkin LM, Irvine DR, Nelson JE, Merzenich MM, Clarey JC.,(1986) Frequency representation in the auditory midbrain and forebrain of a marsupial, the northern native cat (Dasyurus hallucatus). Brain Behav. Evol., 2917-28

Allen R.E. Concise Oxford Dictionary of Current English. Oxford: Clarendon Press, 1990
Baselt R.C., (1987) Salicylate. Analytical procedures for therapeutic drug mornitoring and emergency toxicology 2nd edition, 256-262

Bauer C.A., Brozoski T.J., Rojas P., Boley J., Wyder,M., (1999) Behavioral model of chronic tinnitus in rats. Otolaryngol. Head Neck Surg. 121, 457-462

Bauer CA, Brozoski TJ, Holder TM, Caspary DM., (2000) Effects of chronic salicylate on GABAergic activity in rat inferior colliculus. Hear. Res. 147, 175-182

Bauer C.A., Brozoski T.J., (2001) Assessing tinnitus and prospective tinnitus therapeutics using a psychophysical animal model. J. Assoc. Res. Otolaryngol. 2, 54-64

Carretta D., Hervé-Minvielle A., Bajo V.M., Villa A.E., Rouiller E.M., (1999) c-fos expression in the auditory pathways related to the significance of acoustic signals in rats performing a sensorymotor task. Brain Res 841,170–183

Cazals Y., Li X.Q., Aurousseau C., Didier A., (1988) Acute effects of noradrenalin related vasoactive agents on the ototoxicity of aspirin: An experimental study in the guinea pig. Hear. Res. 36, 89-96

Cazals Y., (2000) Auditory sensori-neural alterations induced by salicylate. Prog. Neurobiol. 62(6), 583-631

Chen G., Jastreboff P.L., (1995) Salicylate induced abnormal activity in the inferior colliculus of rats. Hear. Res. 82, 158-178

Chermak G.D., Dengerink J.E., (1987) Characteristics of temporary noise-induced tinnitus in male and female subjects. Scand. Audiol. 16, 67-73

Coudray C., Mangournet C., Bouhadjeb S., Faure H., Favier A., (1996) Rapid high-performance liquid chromatographic assay for salicylic acid in plasma without solvent extraction. J. Chromatogr. Sci. 34 (4), 166-173

Day R.O., Graham G.G., Bieri D., Brown M., Cairns D., Harris G., Hounsell J., Platt-Hepworth S., Reeve R., Sambrook P.N., (1989) Concentration-response relationships for salicylate-induced ototoxicity in normal volunteers. Br. J. Clin. Pharm. 28, 695-702

Didier A., Miller J.M., Nuttall A.L., (1993) The vascular component of sodium salicylate ototoxicity in the guinea pig. Hear. Res. 69, 199-206

Dragunow M., Faull R., (1989) The use of c-fos as a metabolic marker in neuronal pathway tracing, J. Neurosci. Methods 29, 261–265

Eggermont J., (1990) On the pathophysiology of tinnitus: a review and a peripheral model. Hear. Res. 48, 111-123

Ehret G., Fischer R., (1991) Neuronal activity and tonotopy in the auditory system visualized by c-fos gene expression. Brain Res., 567-354

Ellenhorn M.J., (1997) Ellenhorn’s Medical Toxicology: Diagnosis and Treatment of Human Poisoning, 2nd ed. Baltimore: Williams & Wilkins, 210-223

Fitzgerald, J.J., Robertson, D., Johnstone, B.M., (1993) Effects of intra-cochlear perfusion of salicylates on cochlear microphonic and other auditory responses in the guinea pig. Hear. Res. 67,147-156

Friauf E., (1995) c-fos immunocytochemical evidence for acoustic pathway mapping in rats. Exp. Brain Res. 66, 217–224

Guitton M.J., Caston J., Ruel J., Johnson R.M., Pujol R.M., Puel J.L., (2003) Salicylate induces tinnitus through activation of cochlear NMDA receptors. J. Neurosci. 23, 3944-3952

Heffner H.E., Harrington I.A., (2002) Tinnitus in hamsters following exposure to intense sound. Hear. Res. 170, 83-95

Heller M.F., Bergman M., (1953) Tinnitus aurium in normally hearing persons. Ann. Otol Rhinol. Laryngol. 62, 73–83

Huang J.L., Morgan D.J., (1993) Simple direct injection high-performance liquid chromatographic method to determine quinidine in plasma J. Chromatogr., 620 (2), 278-80

Huffman R.F., Henson O.W., (1990) The descending auditory pathway and acoustic motor systems: connections with the inferior colliculus. Brain Res. Rev. 15, 295-323

Hunt S.P., Williams S., Pini A., Errington M., Bliss T., Evan G., (1989) Visualization of Brain Functions, Macmillan Press, London, 319-329

Jastreboff P.J., Sasaki C.T., (1986)
Salicylate-induced changes in spontaneous activity of single units in the inferior colliculus in the guinea pig. J. Acoust. Soc. Am. 80, 1384-1391

Jastreboff P.J., Brennan J.F., Sasaki, C.T., (1988a) An animal model for tinnitus. Laryngoscope 98, 280-286

Jastreboff P.J., Issing W., Brennan J.F., Sasaki C.T. (1988b) Pigmentation, anesthesia, behavioral factors, and salicylate uptake. Arch. Otolaryngol. Head Neck Surg. 114, 186-191

Jastreboff P.J., Brennan J.F., Colemam J.K., Sasaki, C.T. (1988c) Phantom auditory sensation in rats: An animal model for tinnitus. Behav.Neurosci. 102, 811-822

Jastreboff P.J., (1990) Phantom auditory perception (tinnitus): mechanisms of generation and perception. Neurosci. Res. 8, 221-254

Jastreboff P.J., Brennan, J.F., (1994) Evaluating the loudness of phantom auditory perception (tinnitus) in rats. Audiology 33, 202-217

Jastreboff P.J., Sasaki C.T., (1994) An animal model of tinnitus: A decade of development. Am. J. Oto.15 (1), 19-27

Jastreboff P.J., Gray W.C., Gold S.L. (1996) Neurophysiological approach to tinnitus patients. Am. J. Otol.17, 236-240

Jastreboff P.J., Jastreboff M.M., (2000) Tinnitus Retraining therapy (TRT) as a method for treatment of tinnitus and hyperacusis patients. J. Am. Acad.Audiol. 11,162-177

Jung TT, Miller SK, Rozehnal S, Woo HY, Park YM, Baer W., (1992) Effect of round window membrane application of salicylate and indomethacin on hearing and levels of arachidonic acid metabolites in perilymph. Acta Otolaryngol Suppl. 493, 81-87

Jung T.T., Rhee C.K., Lee C.S., Park Y.S., Choi D.C., (1993) Ototoxicity of salicylate, non-steroidal anti-inflammatory drugs, and quinine. Otolaryngol Clin North Am. 26(5), 791-810

Karnes HT, Shui G., Shah VP, (1991) Validation of bioanalytical methods. Pharm. Res.8 (4), 421-426

Katzung B.G., (1998) Basic and Clinical Pharmacology (7th edition), Appleton & Lange, Norwalk, CN. 578-585

Kaltenbach J.A., (2000) Neurophysiologic mechanisms of tinnitus. J. Am. Acad. Audiol. 11(3), 125-137

Kennedy HJ, Evans MG, Crawford AC, Fettiplace R., (2003) Fast adaptation of mechanoelectrical transducer channels in mammalian cochlear hair cells. Nat. Neusci. 6(8), 832-836

Kenmochi M., Eggermont J.J., (1997) Salicylate and quinine affect the central nervous system. Hear. Res. 113, 110-116

Kees F., Jehnich D., Grobecker H., (1996) Simultaneous determination of acetylsalicylic acid and salicylic acid in human plasma by high-performance liquid chromatography.
J. Chromatogr. B 677, 172-177

King R.C., Miller-Stein C., Magiera D.J., Brann J., (2002) Description and validation of a staggered parallel high performance liquid chromatography system for good laboratory practice level quantitative analysis by liquid chromatography/tandem mass spectrometry. Rapid Commu. Mass Spectrom.16(1), 43-52

Klimeš J., Sochor J., Zahradnicek M., Sedlacek J., (1992) Simultaneous high-performance liquid chromatographic determination of salicylates in whole blood, plasma and isolated erythrocytes. J. Chromatogr. 584, 221-228

Krivosikova Z., Spustova V., Dzurik R., (1996) A highly sensitive HPLC method for the simultaneous determination of acetylsalicylic, salicylic and salicyluric acids in biological fluids: Pharmacokinetic, metabolic and monitoring implications. Methods Find. Exp. Clin. Pharmacol.18(8), 527-532

Kujawa S.G., Fallon M., Bobbin R.P., (1992) Intracochlear salicylate reduces low-intensity acoustic cochlea microphonic distortion products. Hear. Res. 64, 73-80

Kwong T.C., (1987) Salicylate measurement: Clinical Usefulness and Methodology. CritRev. Clin. Lab. Sci. 25, 137–159

Le Prell C.G., Shore S.E., Hughes L.F., Bledsoe S.C. Jr., (2003) Disruption of lateral efferent pathways: functional changes in auditory evoked responses. J Assoc Res Otolaryngol. 4(2), 276-290

Liao P.C., Li C.M., Lin L.C., Hung C.W., (2002) An online automatic sample cleanup system for the quantitative detection of the benzene exposure biomarkers S-phenylmercapturic acid in human urine by electrospray Ionization tandem mass spectrometry. J. Anal. Toxicol. 26, 205-210

Liberman M.C., (1990) Effects of chronic cochlear de-efferentation on auditory-nerve response. Hear Res. 49, 209-223

Lin L.C., Tyan Y.C., Shih T.S., Chang Y.C., Liao P.C., (2004) Development and validation of an isotope-dilution electrospray ionization tandem mass spectrometry method with an on-line sample clean-up device for the quantitative analysis of the benzene exposure biomarker S-phenylmercapturic acid in human urine. Rapid Commun. Mass Spectrom.18, 1310-1316

Liu J.H., Smith P. C., (1996) Direct analysis of salicylic acid, salicyl acyl glucuronide, salicyluric acid and gentisic acid in human plasma and urine by high-performance liquid chromatography. J. Chromatogr. B. 675 (1), 61-70

Lobarinas E., Sun W., Cushing R., Salvi R., (2003) A novel behavioral paradigm for assessing tinnitus using schedule-induced polydipsia avoidance conditioning (SIP-AC). Hear. Res.190, 109-114

Manabe Y, Yoshida S, Saito H, Oka H., (1997) Effects of lidocaine on salicylate-induced discharge of neurons in the inferior colliculus of the guinea pig. Hear. Res. 103, 192-198

McFadden D., Plattsmier H.S., Pasanen E.G., (1984) Aspirin-induced hearing loss as a model of sensorineural hearing loss. Hear. Res.16, 251-260

McMahon G.P., Kelly M.T., (1998) Determination of aspirin and salicylic Acid in human plasma by column-switching liquid chromatography using on-line solid phase extraction. Anal. Chem. 70, 409-414

Medina J.E., Drescher D.G., (1981) The amino-acid content of perilymph and cerebrospinal fluid from guinea-pigs and the effect of noise on the amino-acid composition of perilymph. Neuroscience 6(3), 505-509

Morgan J.I., Curran T., (1991) Stimulus-transcription coupling in the nervous system: involvement of the inducible proto-oncogenes fos and jun. Annu. Rev. Neurosci.14, 421–451

Muller M., Klinke R., Arnold W., Oestreicher E.,(2003) Auditory nerve fiber responses to salicylate revisited. Hear. Res. 183, 37-43

Myers E.M., Bernstein J.M., (1965) Salicylate and ototoxicity. Arch. Otolaryngol. 82, 483-493

Ochi K., Eggermont J.J., (1996) Effects of salicylate on neural activity in cat primary auditory cortex. Hear. Res. 95, 63–76

O'Kruk R.J., Adams M.A., Philp R.B., (1984) Rapid and sensitive determination of acetylsalicylic acid and its metabolites using reversed-phase high-performance liquid chromatography. J. Chromatogr. B, 310, 343-352

Oliver D., He D.Z., Klocker N., Ludwig J., Schulte U., Waldegger S., Ruppersberg J.P., Dallos P., Fakler B., (2001) Intracellular anions as the voltage sensor of prestin, the outer hair cell motor protein. Science 292, 2340-2343

Paxinos G., Watson C., (1986) The Rat Brain in Stereotaxic Coordinates. Academic Press, New York

Peng G.W., Gadalla M.A., Smith V., Peng A., Chiou W.L., (1978) Simple and rapid high-pressure liquid chromatographic simultaneous determination of aspirin, salicylic acid, and salicyluric acid in plasma. J. Chromatogr. 67, 710-712

Penner M.J., Jastreboff P.J. (1996) Tinnitus: psychophysical observations in humans and an animal model. In: van de Water, T.R., Popper, A.N., Fax, R.R. (Eds.), Clinical Aspects of Hearing. Springer, New York, pp. 258-304

Puel J.L. Bobbin R.P., Fallon M., (1990) Salicylate, mefenamate, meclofenamate, and quinine on cochlea potential. Arch. Otolaryngol. Head Neck Surg. 102, 66-73

Rainsford K.D., (1984) Aspirin and salicylates. Butterworths, Boston.P

Rumble R.H., Roberts M.S., (1981) Determination of aspirin and its major metabolites in plasma by high-performance liquid chromatography without solvent extraction.
J. Chromatogr., 225, 252-260

Sagar S.M., Sharp F.R., Curran T., (1988) Expression of c-fos protein in brain: metabolic mapping at the cellular level. Science 240, 1328-1331

Seligmann H, Podoshin L, Ben-David J, Fradis M, Goldsher M., (1996) Drug-induced tinnitus and other hearing disorders. Drug Saf. 14(3), 198-212

Shen J., Wanwimolruk S., Clark C.R., Roberts M.S., (1990) A sensitive assay for aspirin and its metabolites using reversed-phase ion-pair high-performance liquid chromatography. J. Liq. Chromarogr.13 (4), 751-761

Sheng M., Greenberg M.E., (1990) The regulation and function of c-fos and other immediate early genes in the nervous system. Neuron 4(4), 477-485

Souverain S., Rudaz S., Veuthey J.L., (2004) Restricted access materials and large particle supports for on-line sample preparation: an attractive approach for biological fluids analysis. J. Chromatogr. B 801(2), 141-156

Stewart M.J., Watson I.D., (1987) Analyrical reviews in clinical chemistry: methods for the estimation of salicylate and paracetamol in serum, plasma, and urine. Ann. Clin. Biochem. 24, 552-565

Takano T., Hata S.J., (1996) High-performance liquid chromatographic determination of finasteride in human plasma using direct injection with column switching. J. Chromatogr. B, 676 (1), 141-146

Thalmann R., (1992) Protein profiles of perilymph and endolymph of the guinea pig. Hear Res. 63, 37-42

Trinder P., (1954) Rapid determination of salicylate in biological fluids. J. Biochem. 57, 301-303

Wallhäusser-Franke E., Braun S, Langner G., (1996) Salicylate alters 2-DG uptake in the auditory system: a model for tinnitus? NeuroReport 7, 1585-1588

Wallhäusser-Franke E., (1997) Salicylate evokes c-fos expression in the brain stem: implications for tinnitus. NeuroReport 8, 725-728

Wersall J., (1995) Ototoxic antibiotics: a review. Acta Otolaryngol Suppl. 519, 26-29

Wu J.L., Chiu T.W., Poon P. W. F., (2003) Differential changes in Fos-immunoreactivity at the auditory brainstem after chronic injections of salicylate in rats. Hear. Res. 176, 80-93

Veuthey J.L., Souverain S., Rudaz S., (2004) Column-switching procedures for the fast analysis of drugs in biologic samples. Ther. Drug Monit. 26(2), 161-166

Yan W., Suga N., (1998) Corticofugal modulation of the midbrain frequency map in the bat auditory system. Nat. Neurosci. 1, 54-58

Yu Z., Westerlund D., (1996) Direct injection of large volumes of plasma in a column-switching system for the analysis of local anaesthetics. II. Determination of bupivacaine in human plasma with an alkyldiol silica precolumn. J Chromatogr A. 725(1), 149-155

Xu, J.F., Poon, P.W.F., Chen, X.Y., Chung, S.N., (1990) Computerbased three dimensional reconstructions of nuclear subdivisions in the inferior colliculus. Comput. Engin. Appl. (Chinese) 6, 7-10
論文全文使用權限
  • 同意授權校內瀏覽/列印電子全文服務,於2004-08-25起公開。
  • 同意授權校外瀏覽/列印電子全文服務,於2004-08-25起公開。


  • 如您有疑問,請聯絡圖書館
    聯絡電話:(06)2757575#65773
    聯絡E-mail:etds@email.ncku.edu.tw