Chung Hwa University of Medical Technology
Chung Hwa University of Medical Technology RepositoryNot a member yet
1453 research outputs found
Sort by
Signal amplification and optimization of aptamer microarray by rolling circle amplification
Get PDF目 錄
口試委員會審定書
授權書 ii
中文摘要 iii
英文摘要 iv
誌謝 v
第一章、 緒論
1-1研究背景 -1-
1-2 研究目的與動機 -2-
1-3 論文架構 -4-
第二章、 理論分析與探討
2-1 生物晶片之起源 -5-
2-2 生物晶片之種類 -6-
2-3 抗原/抗體反應 -8-
2-4 人類免疫球蛋白E -9-
2-5 鏈菌素與生物素之結合作用 -10-
2-6生物晶片之載台製作 -10-
2-7 相轉換法 -11-
2-8 滾動循環擴增技術 -13-
2-9 適合體 -15-
2-10訊號呈現 -16-
第三章、 實驗方法與材料
3-1 材料 -17-
3-2晶片表面處理 -20-
3-3電泳 -20-
3-4 儀器與器材 -20-
3-5 溶液製備 -21-
3-6 實驗步驟 -22-
3-6.1 生物晶片之製備 -22-
3-6.2 滾動循環擴增技術 -25-
3-6.3 環狀模板之製備 -25-
3-6.4 環狀模板分子成形之驗證 -27-
3-6.5 Agarose gel之配製 -27-
3-6.6 液相RCA驗證 -28-
3-6.7 模板是否與引子互補經RCA之影響(微量離心管) -29-
3-6.8 模板是否與引子互補經RCA之影響(生物晶片) -30-
3-6.9 驗證適合體結合Human IgE之能力 -31-
3-6.10驗證適合體結合Human IgE經RCA放大 -33-
3-6.11 驗證蛋白質之非特異性放大 -34-
3-6.12驗證適合體結合Human IgG之能力 -35-
3-6.13驗證適合體結合Human IgG經RCA放大 -37-
3-6.14 不同濃度適合體結合Human IgE比較其螢光強度 -38-
3-6.15 改變RCA反應時間探討其最佳化反應時間 -39
第四章、 結果與討論
4-1環狀模板分子成形之驗證 -40-
4-2液相RCA驗證 -41-
4-3模板是否與引子互補經RCA之影響(微量離心管) -42-
4-4模板是否與引子互補經RCA之影響(生物晶片) -43-
4-5驗證適合體結合Human IgE之能力 -44-
4-6驗證適合體結合Human IgE經RCA結果 -45-
4-7驗證蛋白質之非特異性放大 -46-
4-8驗證適合體結合Human IgG之能力 -46-
4-9驗證適合體結合Human IgG經RCA結果 -47-
4-10不同濃度適合體結合Human IgE比較其螢光強度 -48-
4-11改變RCA反應時間探討其最佳化反應時間 -48-
4-12 綜合討論 -49-
第五章、結論 -52-
第六章、參考文獻 -53-
附錄 -60-
參考文獻
Ellington A.D., Szostak J.W.(1990) In vitro selection of RNA moleculesthat bind specific ligands, Nature. 346 ,818–822.
Ellington A.D., Szostak J.W. (1992) Selection in vitro of single-stranded
DNA molecules that fold into specific ligand-binding structures , Nature .355, 850–852.
Schweitzer B. , Wiltshire S. , Lambert J. , Malley S.O., Kukanskis K., Zhu Z., Kingsmore S. F., Lizardi P.M., andWard D.C. (2000) , Immunoassays with rolling circle DNA amplification: A versatile platform for ultrasensitive antigen detection. PNAS ,97,10113-10119.
Tuerk C., Gold L. (1990) Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase, Science. 249,505–510.
Lauhon C., Szostak J. (1993) In vitro selection of RNA molecules that
specifically bind riboflavin, FASEB J. 7, A1087.
Huizenga D.E., Szostak J.W. (1995) A DNA aptamer that binds adenosine and ATP, Biochemistry .34, 656–665.
Bunka D.H., Stockley P.G. (2006) Aptamers come of age—at last, Nat.
Rev. Microbiol. 4,588–596.
Wilson D.S., Szostak J.W. (1999) In vitro selection of functional nucleic
acids, Annu. Rev. Biochem. 68,611–647.
Dobrovolskaia E.; Gam A.;SlaterJ. E . (2006),Competition enzyme-linked immunosorbant assay (ELISA) can be a sensitive method for the specific detection of small quantities of allergen in a complex mixture., Clinical and Experimental Allergy, 36,525-530.
Lin F. C., Wang D. M. and Lai J. Y.. (1996). Asymmetric TPX membranes with high gas flux. J. Membr. Sci., 110, 25.
Hsu H.Y, Huang Y.Y (2004), RCA combined nanoparticle – based optical detection technique for protein microarray:a novel approach, Biosensors and Bioelectronics ,20,123-126.
Tok J.B., Cho J., Rando R.R. (2000) RNA aptamers that specifically bind
to a 16S ribosomal RNA decoding region construct, Nucleic Acids
Res. 28, 2902–2910.
Kuby J. (1997) Immunology, 3rd ed. , New York, 1997(Chapter 5).
Ho J.A,Wu L.C, Chang L.H, Hwang K.C, Hwu J.R (2010) Liposome- based immunoaffinity chromatographic assay for the quantitation of immunoglobulin E in human serum, Journal of Chromatography B,2,172-176.
Lai J. Y., Liu M. J. and Lec K. R. (1994). Polycarbonate membrane prepared via a wet phase inversion method for oxygen enrichment from air. J. Membr.Sci., 86, 103.
Lai J. Y., Lin S. F., Lin F. C. and Wang D. M.. (1998). Constructure of ternary phase diagrams in nonsolvent/solvent/PMMA systems. J. Polym. Sc., Part B：Polym. Phys., 36, 607.
Cheng J.Y, Hsieh C.J, Chuang Y.C and Hsieh J.R. (2005) ,Performing microchannel temperature cycling reactions using reciprocating reagent shuttling along a radial temperature gradient.Analyst.130, 931-940
Joerger and Hendrickson(1995)Analyte detection with DNA-labeled antibodies and polymerase chain reaction. Clinical Chemistry,41,1371-1377.
Kesting R. E. (1773) Concerning the microstructure of dry-RO membrane, J.Appl. Polym. Sci., 17, 1771.
Kurata M. (1982). Thermodynamics of polymer solutions. Harwood Academic,London.
Ishizaka K., Ishizaka T., Hornbrook J.M.M., (1967) Allergen-Binding Activity of E, G and A Antibodies in Sera from Atopic Patients: In Vitro Measurements of Reaginic Antibody, J. Immunol. 98, 490-501
Kesting R. E. (1773). Concerning the microstructure of dry-RO membrane, J.Appl. Polym. Sci., 17, 1771.
Kurata M. (1982). Thermodynamics of polymer solutions. Harwood Academic,London.
Lloyd D. (1990). Microporous membrane formation via thermally induced phase separation. I. Solid-liquid phase separation. J. Membr. Sci., 52, 239.
Koizumi M., Breaker R.R. (2000) Molecular recognition of cAMP by an
RNA aptamer, Biochemistry.39, 8983–8992.
Mehta R. H., Madsen D. A. and Kalika D. S. (1995). Microporous membranes based on poly(ether ether ketone) via thermally-induced phase separation.J.Menbr. Sci., 107, 93.
Mulder M. (1991). Basic principle of membrane technology. Kluwer
Academic Publisher, London.
Porschewski P., Grattinger M.A., Klenzke K., Erpenbach A., Blind M.R., Schafer F. (2006) Using aptamers as capture reagents in bead-based
assay systems for diagnostics and hit identification, J. Biomol. Screen.
11,773–781
Jayasena S.D. (1999) An emerging class of molecules that rival
antibodies in diagnostics, Clin. Chem. 45,1628-1650.
S.G.O.Johansson,(1967) RAISED LEVELS OF A NEW IMMUNOGLOBULIN CLASS (IgND) IN ASTHMA, The Lancet,290,951-953.
Strathmann H. and Kock K. (1977). The formation mechanism of phase
inversion membrane. Desalination, 21,241.
Tarasow T.M., Tarasow S.L., Eaton B.E. (1997) RNA-catalysed carbon–
carbon bond formation, Nature. 389,54-57.
Vandeweerdt P. and Berghmans H. (1991). Temperature-coccentration
behavior of solution polydisperse, atactic poly(methyl methacrylate) and its influence on the formation of amorphous, micropous membranes. Macromolecules, 24, 3547.
Shao W., Zhou Z., Laroche I., Lu H., Zong Q., Patel D.D., Kingsmore S., and P.Piccoli S.. (2003) Optimization of Rolling-circle Amplified Protein Microarray for Multiplexed Protein Profiling. Journal of Biomedicine Biotechnology,5,299-307.
Yanagishita H., Nakane T. and Yoshitome H. (1994). Selection criteria for solvent and gelation medium in the phase inversion process. J. Membr. Sci.,89, 215.
Zeman L. and Fraser T. (1993). Formation of air-cast cellulose acetate
membranes. Part II. Kinetic of demixing and macrovoid growth, J. Membr.Sci., 87, 267.
Zsigmongy R. and Bachman W. (1992). Filter and method for producing same.US Patent 1,421,341.
林明泉。 1999。臨床血清免疫學 第二版。
呂偉立。DNA 型式碳水化合物抗原之生物偵測器的開發及應用。2004。國立中央大學化學研究所碩士論文。
葉庭秀。利用滾動循環擴增技術之細胞激素陣列化螢光分析訊號放大與最適化探討。2007。中原大學化學工程學系碩士學位論文。
吳炳賢。基因微陣列在濕阻證病理造型研究上之應用。2003。中國醫藥學院中國藥學研究所碩士論文。
黃朝裕。壓印式蛋白質微陣列晶片之印台研發。1991。國立清華大學工程與系統科學所微機電組碩士論文。[[abstract]]適合體是一種單股核酸序列，其會在特殊環境狀態下折疊形成四級結構。當適合體折疊形成此種特殊結構時即會具備高特異性、敏感性結合標的分子的能力。適合體所能結合的標的分子主要包括蛋白質、小分子、DNA及RNA。滾動循環擴增技術(Rolling circle amplification，RCA)是一種核酸放大的技術，此種技術可應用於蛋白質檢測上。傳統的免疫分析方式結合RCA(Immuno-RCA)的檢測技術，此種技術有一個主要的缺點即是需建構出antibody-DNA hybrid之架構且建構出此hybrid具有困難性且耗時，因此在本論文中我們藉由Aptamer-DNA之架構去改善Immuno-RCA的缺點。適合體在於使用上有幾項優點為(1)研發容易，(2)穩定性高，(3)合成容易，(4)用途廣泛。在此我們利用適合體結合RCA技術( Aptamer- RCA )發展出一種高敏感性且高特異性檢測蛋白質的技術。本論文中，我們以人類免疫球蛋白E( Human IgE )作為標的蛋白，發展出過敏原晶片檢測系統，並證實適合體結合RCA技術是一種高敏感性、特異性，並可達廣泛的檢測範圍。因適合體可廣泛應用在於多種蛋白質檢測上，因此將此種平臺應用在於過敏檢測上將具有很好的願景。
Aptamers are single-stranded nucleic acids(ss DNA or ss RNA) that are able to fold into defined tertiary structures. If aptamer fold into the special structure, it will have high specificity and affinity while binding with target molecular. Targets to which aptamers have been generated against successfully include proteins, small molecules, and RNA. A different nucleic acid amplification technique called rolling circle amplification (RCA),is adapted to the detection of proteins. A defect of immuno-RCA is that the hybrids can be problematic and time-consuming, so we used aptamer-DNA to improve this problem. Aptamers have some advantages: (1) easy to development, (2) stable, (3) robust methods for their synthesis, and (4) many extensive applications. Aptamer-based rolling circle amplification (aptamer-RCA) is a kind of high sensitivity and high specificity method for the detection of protein. In this research, we utilized human IgE as a model target to develop allergy biochip. And it was demonstrated that the aptamer-RCA presented method was highly sensitive and specific with a broaden detection range and a low detection limit. Because of the wide availability of aptamers for multiple proteins, this platform holds great promise in allergy examination
Immobilization of enzyme on nitrocellulose film for pH-EGFET type biosensors
Get PDF口試委員審定書
授權書 ii
中文摘要 iv
英文摘要 v
誌謝 vi
第一章 緒論
1.1 研究背景 1
1.2 研究目的與動機 4
1.3 論文架構 6
第二章 理論分析與探討
2.1 生物感測器之種類與應用 8
2.2 酵素固定化技術 11
2.3 延伸式閘極感測場效電晶體之原理 13
2.4 相轉換法（Phase inversion process） 15
2.5 成膜理論 18
2.6 尿素之生理作用機制 22
2.7 尿素循環 25
第三章 材料方法
3.1 實驗材料 26
3.2 實驗流程 27
3.3 感測器之製作 28
3.4 量測系統 29
3.5 硝化纖維素之表面活化 30
3.6 尿素感測器之酵素固定化作用 31
第四章 結果與討論
4.1 硝化纖維素溶液之phase inversion process 32
4.2 硝化纖維素的濃度與旋轉塗佈轉速會影響pH感測度 34
4.3 尿素感測器之反應電壓 35
第五章 結論 36
參考文獻 37
圖表目錄
圖1-1、生物感測器之原理 44
圖1-2、Scheme of the second measurement setup with the microreactor packed with lipase immobilised onto the NC sheets and lipase coupled to glass beads coated with keratin. 45
圖1-3、論文架構圖 46
圖2-1、電化學生物感測器之種類 47
圖2-2、硫酸溶液的氧化還原反應 49
圖2-3、pH Optical sensor 50
圖2-4、QCMs(quartz-crystal microbalances ) 51
圖2-5、MOSFET、ISFET、ENFET之構造 52
圖2-6、離子感測場效電晶體(ISFET) 53
圖2-7、酵素固定化方式(a)吸附(b)微粒包覆(c)包埋(d)交聯(e)共價鍵結 54 圖2-8、延伸式閘極感測場效電晶體（EGFET）之結構圖 55
圖2-9、非對稱離子選擇元件之等效電路 56圖2-10、Schematic representation of a ternary phase diagram of polymer/solvent/nonsolvent 57
圖2-11、Schematic repressentation of casting film/coagulant interface 58
圖2-12、尿素循環 59
圖3-1、實驗流程圖 60
圖3-2、SnO2/ITO玻璃元件基本架構 61
圖3-3、測量系統 62
圖3-4、硝化纖維素溶液配製流程 63
圖3-5、尿素感測器之酵素固定化 64
圖4-1、不同比例的去離子水於硝化纖維素溶液中其元件之感測度 65
圖4-2、添加不同比例去離子水之硝化纖維素溶液之SEM 66
圖4-3、不同濃度的硝化纖維素修飾於延伸式閘極感測場效電晶體表面之感測度 67
圖4-4、不同轉速塗佈修飾硝化纖維膜之元件其pH感測度 68
圖4-5、尿素反應曲線圖 69
圖4-6、尿素濃度64 mg/dl之反應電壓 70
圖4-7、尿素感測器校正曲線圖 71
圖4-8、尿素感測器穩定度測試 72
參考文獻
Abeyrathne PD, Lam JS. (2007). Conditions that allow for effective transfer of membrane proteins onto nitrocellulose membrane in Western blots. Can. J. Microbiol. 53, 526-532.
Ahmad A.L., Low S.C., Shukor S.R.Abd. (2007). Effect of membrane cast thickness on controlling the macrovoid structure in lateral flow nitrocellulose membrane and determination of its characteristics. Scripta Materialia 57, 743-746.
Bergveld P. (1970). Development of an ion sensitive solid-state device for neurophysiological measurements. IEEE Transactions on Bio-medical Engineering 17, 70-71.
Boubriak O.A., Soldatkin A.P., Starodub N.F., Sandrovsky A.K., El’skaya A.K. (1995). Determination of urea in blood serum by a urease biosensor based on an ion-sensitive filed-effect transistor. Sensors and actuators B, 26-27, 429-431.
Carles Puig-Lleixà, Cecília Jiménez, Julián Alonso, Jordi Bartrolí. (1999). Polyurethane-acrylate photocurable polymeric membrane forion-sensitive field-effect transistor based urea biosensors. Analytica Chimica Acta 389, 179-188.
Charles M. Barksdale. (1998). On-probe immunoaffinity extraction by matrix-assisted laser desorption/ionization mass spectrometry, Analtical Chemistry 70, 498-503.
Chung-We P., Jung-Chuan C., Tai-Ping S., Shen-Kan H. (2005). Development of the tin oxide pH electrode by the sputtering method. Sensors and Actuators B, 108, 863-869.
Clark L. C., C. Lyois.(1962). Electrode system for continuous monitoring in cardiovascular surgery. Annals of the New York Academy of Sciences 102, 29-33.
Flory P. J. (1953). Principles of polymer chemistry. Cornell University Press. New York.
F. C. Lin, D. M. Wang and J. Y. Lai. (1996). Asymmetric TPX membranes with high gas flux. J. Membr. Sci., 110, 25.
Gershoni J.M., Palade G.E. (1983). Protein blotting: principles and applications. Anal. Biochem. 131, 1-15.
Janta J. (1983). Electrochemistry of chemically sensitive field effect transistors. Sensors and Actuators B, 4, 255-265.
J. Y. Lai, M. J. Liu and K. R. Lec. (1994). Polycarbonate membrane prepared via a wet phase inversion method for oxygen enrichment from air. J. Membr. Sci., 86, 103.
J. Y. Kim, H. K. Lee, K. J. Bank and S. C. Kim. (1997). Liquid-liquid phase separation in polysulfone/solvent/water systems. J. Appl. Polym. Sci., 65,2643.
J. Y. Lai, S. F. Lin, F. C. Lin and D. M. Wang. (1998). Constructure of ternary phase diagrams in nonsolvent/solvent/PMMA systems. J. Polym. Sc., Part B：Polym. Phys., 36, 607.
Jia-Chyi Chen, Jung-Chuan Chou, Tai-Ping Sun, Shen-Kan Hsiung. (2003). Portable urea biosensor based on the extended-gate field effect transistor. Sensors and actuators B, 91, 180-186.
Jun-Qi Wang, Jung-Chuan Chou, Tai-Ping Sun, Shen-Kan Hsiung, Guang-Bin Hsiung. (2003). pH-based potentiometrical flow injection biosensor for urea. Sensors and actuators B, 91, 5-10.
Kesting R. E. (1773). Concerning the microstructure of dry-RO membrane, J. Appl. Polym. Sci., 17, 1771.
Kurata M. (1982). Thermodynamics of polymer solutions. Harwood Academic, London.
Katsube T., Araki T., Hara M., Yaji T., Si Kobayashi and K. Suzuki. (1986). A multi-species biosensor with extended-gate field effect transistors. Proceeding of 6th Sensor Symposium, Tsukuba Japan, 211-214.
Kailasapathy K., Chin J. (2000). Survival and therapeutic potential of probiotic organisms with reference to Lactobacillus acidophilus and Bifidobacterium spp., Immunology and Cell Biology 78, 80-88.
Lloyd D. (1990). Microporous membrane formation via thermally induced phase separation. I. Solid-liquid phase separation. J. Membr. Sci., 52, 239.
Li-Te Yin, Jung-Chuan Chou, Wen-Yaw Chung, Tai-Ping Sun and Shen-Kan Hsiung. (1999). Separative structure ISFETs on a glass substrate. in:Proceeding of the International Symposium on Photonics and Applications, Nanyang Technological University, Singapore 3987, 416-419.
Li-Te Yin, Jung-Chuan Chou, Wen-Yaw Chung, Tai-Ping Sun, Shen-Kan Hsiung. (2000). Separate structure extended gate H+-ion sensitive field effect transistor on a glass substrate, Sensors and Actuators 71, 106-111.
Li-Ye Yin, Chih-Yu Hu and Cheng-Hao Chang. (2008). A single layer nitrocellulose substrate for fabricating protein chips. Sensors and Actuators B. 130, 374-378.
Mulder M. (1991). Basic principle of membrane technology. Kluwer Academic Publisher, London.
Mehta R. H., Madsen D. A. and Kalika D. S. (1995). Microporous membranes based on poly(ether ether ketone) via thermally-induced phase separation. J.Menbr. Sci., 107, 93.
Martin Bakker, Fred van de Velde, Fred van Rantwijk, Roger A. Sheldon. (2000). Highly efficient immobilization of glycosylated enzymes into polyurethane foams. Biotechnology and Bioengineering70, 342-348.
Marhall L. Summar, MD. (2003). Urea cycle disorders overviews. Genereviews, April 29.
Nygaard A.P., Hall B.D. (1963). A method for the detection of RNA-DNA complexes. Biochemical and Biophysical Research Communications 12, 98-104.
Pijanowska D.G., Baraniecka A, Wiater R., Ginalska G., Lobarzewski J., Torbicz W. (2001). The pH-detection of triglycerides. Sensors and Actuator B. 78, 263-266.
Rebriiev A.V., Starodub N.F. (2004). Enzymatic biosensor based on the ISFET and photopolymeric membrane for the determination of urea. Electroanalysis 16, 1891-1895.
Rasooly Avraham. (2006). Moving biosensors to point-of-care cancer diagnostics. Biosensors and Bioelectronics 21, 1847-1850.
Southern E.M. (1975). Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of molecular biology 98, 503-517.
Strathmann H. and Kock K. (1977). The formation mechanism of phase inversion membrane. Desalination, 21,241.
Stefan Oehler, Regina Alex, Andrew Barke. (1999). Is nitrocellulose filter binding really a universal assay for protein-DNA interations, Analytical Biochemistry 268, 330-336.
Sant W., M.L., J. Launay, Do Conto T., Martinez A., Temple-Boyer P. (2003). Development of chemical field effect transistors for the detection of urea. Sensors and actuators B, 95, 309-314.
Starodub N. F., Rebriev A.V. (2007). Liquid photopolymerizable compositions as immobilized matrix of biosensors. Bioelectrochemistry 71, 29-32.
Tompa H. (1956). Polymer solutions. Butterworths, London.
Tai-Horng Young, Leo-Wang Chen. (1995). Pore formation mechanism of membranes from phase inversion process, Desalination 103, 233-247.
Pourciel
Van Der Spoegel J., Lauks I., Chan P. and Babic D. (1983). The extended gate chemical sensitive filed effect transistor as multi-species microprobe. Sensors and Actuators B. 4, 291-298.
Vandeweerdt P. and Berghmans H. (1991). Temperature-coccentration behavior of solution polydisperse, atactic poly(methyl methacrylate) and its influence on the formation of amorphous, micropous membranes.Macromolecules, 24, 3547.
Xiaoli Liang, David M. Lubman, David T. Rossi, Gerald D. Nordblom, Charles M. Barksdale. (1998). On-probe immunoaffinity extraction by matrix-assisted laser desorption/ionization mass spectrometry, Analtical Chemistry 70, 498-503.
Yanagishita H., Nakane T. and Yoshitome H. (1994). Selection criteria for solvent and gelation medium in the phase inversion process. J. Membr. Sci., 89, 215.
Zsigmongy R. and Bachman W. (1992). Filter and method for producing same. US Patent 1,421,341.
Zeman L. and Fraser T. (1993). Formation of air-cast cellulose acetate membranes. Part II. Kinetic of demixing and macrovoid growth, J. Membr. Sci., 87, 267.
黃冠勛。探討添加劑對巨型孔洞形成之影響。2002。中原大學化學工程系碩士學位論文。
陳佳琪。可拋棄式尿素感測器與前置放大器之研究。2003。中原大學電子工程系碩士學位論文。
王彥盛。利用酵素固定化方式製備尿素感測器之研究。2004。國立雲林科技大學電子與資訊工程研究所碩士論文。
謝振榮。2006年8月。物理雙月刊，廿十八卷四期，704-710。[[abstract]]本篇研究中，我們首先提出利用旋轉塗佈技術將硝化纖維膜固定於延伸式閘極感測場效電晶體，並將酵素固定於硝化纖維膜上用以探討蛋白質之固定效率。旋轉塗佈所使用的硝化纖維素溶液之濃度將會影響延伸式閘極感測場效電晶體之感測度，因此為了找出表面最佳化的條件且又能維持其感測度與蛋白質固定能力，針對硝化纖維溶液的配製包含固定化基材濃度、溶劑與非溶劑比例與旋轉塗佈的轉速做探討，其結果顯示配製硝化纖維溶液最主要的參數為9%硝化纖維、3%去離子水與3000 rpm之轉速，將此硝化纖維溶液旋轉塗佈於延伸式閘極感測場效電晶體上其感測度仍可維持在53±1.2 mV/pH，此外為了了解延伸式閘極感測場效電晶體經硝化纖維膜修飾後其固定蛋白質能力，我們利用尿素酶固定於經硝化纖維膜修飾的延伸式閘極感測場效電晶體，形成尿素感測器，此尿素感測器於尿素濃度為64 mg/dl之尿素溶液中其反應電壓達136mV，硝化纖維膜表面的多孔性形成一種低擴散障蔽，使此尿素感測器於反應時間約20秒時即達最大反應電壓。尿素感測器穩定度之測試，元件在尿素溶液中反覆量測，量測27次後此酵素感測器仍然可保持其良好的工作性能。與包埋固定法相互對照，硝化纖維膜之固定蛋白質能力相似，但其形成較低的擴散障蔽。
In this paper, we propose an EGFET biosensor which the enzyme was immobilized on the nitrocellulose film. A porous nitrocellulose layer formed by phase inversion process was deposited on SnO2/ITO gate directly by spin-coated technology. To reach a surface chemistry condition can corresponding EGFET has appropriate pH-sensitivity and protein immobilization capability, the optimized nitrocellulose film manufacturing conditions were discussed through factors of inverse phase process, spin speed of the spin coater concentration of nitrocellulose. To understand the protein immobilization ability of nitrocellulose modified EGFET, we immobilized the urease on nitrocellulose film which used as an urea biosensor. The device shows a response of 136mV in a urea concentration of 64mg/dl. The porous surface of nitrocellulose shows low diffusion barrier, which has a fast response time of 20 sec. After a stability test, we found the sensor could maintain its excellent performance for detection of urea samples at least repeat measured of 27 times during 60 days. It is concluded that the developed nitrocellulose modified EGFET is perspective for its biosensor application for the determination enzyme or antigen-antibody reaction that the proposed method to prepare a selective biological membrane
Studies on the GABAA Receptor of Betel Quid Extracts in Male Rat
Get PDF目錄
中文摘要……………………………………………………………… ii
英文摘要…………………………………………………………… iv
致謝………………………………………………………………… v
目錄…………………………………………………………………… vi
縮寫表……………………………………………………………… vii
第一章 緒論………………………………………………… 1
第一節 前言……………………………………………………… 1
第二章 文獻探討
第一節 檳榔及其相關成分…………………………………………4
第二節 檳榔對神經系統的影響……………………………………6
第三節 GABA………………………………………………………8
(一) 中樞神經系統………………………………………… 9
(二) 周邊系統………………………………………………10
第四節 GABAA 受體…………………………………………… 11
第五節 Mitogen-activated protein kinase (MAPK) 訊息傳遞路徑
與細胞凋亡……………………………………………… 12
第六節 十個基因簡介…………………………………………… 15
第三章 目的………………………………………………… 19
第四章 材料與方法………………………………………… 20
第一節 實驗設計…………………………………………………20
第二節 實驗動物…………………………………………………20
第三節 實驗前處理與方法………………………………………21
第四節 組織Total RNA萃取…………………………………….25
第五節 反轉錄PCR分析……………………………………… 26
第六節 即時聚合酶鏈鎖反應……………………………………27
第七節 組織蛋白質萃取…………………………………………29
第八節 西方墨點法分析…………………………………………31
第五章 結果………………………………………………… 35
第六章 討論………………………………………………… 38
第七章 結論………………………………………………… 41
參考文獻…………………………………………………… 42
表與圖……………………………………………………… 48
附錄………………………………………………………… 61
參考文獻
B. J. BOUCHER & N. MANNAN. Metabolic effects of the consumption of Areca catechu(2002). Addiction Biology 7, 103- 110.
Burton-Bradley BG: Papua and New Guinea trans-cultural psychiatry: Some implications of betel chewing. Med J Aust 1960;2:744-6.
Cathy L. Bell-Horner, Akiko Dohi, Quynh Nguyen, Glenn H. Dillon,
Meharvan Singh(2006). ERK/MAPK Pathway Regulates GABAA Receptors. Received 27 February 2006; revised 5 July 2006; accepted 7 July 2006.
Chu NS: Effects of betel chewing on the central and autonomic nervous systems. J Biomedical Science 2001;8:229-36.
Chu NS: Betel chewing increases the skin temperature: effects of atropine and propranolol.Neurosci Lett 1995;194:130-2.
Chu NS: Effect of betel chewing on RR interval variation. J Formos Med Assoc1995; 94: 106-10.
Christopher J. Martyniuka, Rosalie Awada, Rachel Hurleya,
Thomas E. Fingerb, Vance L(2007). Trudeaua, Glutamic acid decarboxylase 65, 67, and GABA-transaminase mRNA expression and total enzyme activity in the goldfish (Carassius auratus) brain.Brainr Esearch 1147 154-166.
Denys V. Volgin , Leszek Kubin(2007).Regionally selective effects of GABA on hypothalamic GABAA receptor mRNA in vitro. Received 30 November 2006.
David F.Owens and Arnold R. Kriegstein. IS THERE MORE TO GABA THANSYNAPTIC INHIBITION?( 2002). VOLUME 3 |SEPTEMBER 715.
Dowse GK. Betel-nut chewing and diabetes in Papua New Guinea
and elsewhere. Diabetologia 1994;37:1062-4.
Hitomi FUJIOKA, Keitaro YAMANOUCHI1, Tatsuo AKEMA2 and
Masugi NISHIHARA1(2007). The Effects of GABA on Embryonic Gonadotropin-Releasing Hormone Neurons in Rat Hypothalamic Primary Culture. Journal of Reproduction and Development, Vol. 53, No. 2.
Haubrich DR, Watson DR: Effect of pilocarpine or arecoline administration on acetylcholine levels and serotonin turnover in rat brain. J Pharmacol Exp Ther 1972;181:19-27.
IARC. Betel quid areca-nut chewing. IARC Monogr Eval Carcinog
Risk Chem Hum 1985;37:137-202.
Ko YC, Chiang TA, Chang SJ, Hsieh SF. Prevalence of betel quid
chewing habit in Taiwan and related sociodemographic factors. J
Oral Pathol Med 1992;21:261-4.
Ko YC, Huang YL, Lee CH, Chen MJ, Lin LM, Tsai CC. Betel quid
chewing, cigarette smoking and alcohol consumption related to oral
cancer in Taiwan. J Oral Pathol Med 1995;24:450-3.
Molinengo L, Fundaro AM, Cassone MC: Action of a chronic arecoline administration on mouse motility and on acetylcholine concentrations in the CNS. J Pharm Pharmacol 1988; 40:821-2.
Mondadore C, Hengerer B, Ducret T, Borkowski J: Delayed emergence of effects of memory-enhancing drugs: implication for dynamics of long-term potentiation. Proc Natl Acad Sci USA 1994; 91: 2041-5.
Muir CS. Kirk R: Betel, tobacco and cancer of the month. Br J Cancer 1960; 14: 597-608.
Mark C. McLeod , Suresh Sundram , Brian Dean (2008) Treatment with haloperidol and diazepam alters GABAA receptor density in the rat brain.Progress in Neuro-Psychopharmacology & Biological Psychiatry 32560–567.
M. J. W. Chang, C. Y. Ko, R. F. Lin, L. L. Hsieh. Biological Monitoring of Environment Exposure to Safrole and the Taiwanese
Betel Quid Chewing (2002). Arch. Environ. Contam. Toxicol. 43, 432–437.
Mannan N, Boucher BJ, Evans SJ. Increased waist size and weight in
relation to consumption of Areca catechu (betel-nut); a risk factor for
increased glycaemia in Asians in east London. Br J Nutr 2000;83: 267
-75.
Mashberg A, Boffetta P, Winkelman R, Garfinkel L. Tobacco
smoking, alcohol drinking, and cancer of the oral cavity and oropharynx among U.S. veterans. Cancer 1993;72:1369-75.
Marshall JR, Graham S, Haughey BP, et al. Smoking, alcohol,
dentition and diet in the epidemiology of oral cancer. Eur J Cancer B
Oral Oncol 1992;28B:9-15.
Nelson BS, Heischober B: Betal nut: A common drug used by naturalized citizens from India, Far East Asia and the South Pacific Islands. Ann Emerg Med 1999;34: 238-43.
N.G. Bowery & T.G. Smart(2006) GABA and glycine as neurotransmitters: a brief history. British Journal of Pharmacology 147, S109–S119.
Olga A. Sergeeva, Nadja Andreeva, Maurice Garret, Annette Scherer, and Helmut L. Haas(2005). Pharmacological Properties of GABAA Receptors in RatHypothalamic Neurons Expressing the ε-Subunit .The Journal of Neuroscience, January 5, 2005 • 25(1):88 –95.
Raffaele KC, Berardi A, Asthana S, Morris P, Haxby JV, Soncrant TT: Effects of long-term continuous infusion of the muscarinic cholinergic agonist arecoline on verbal memory in dementia of the Alzheimer type. Psychopharmacol Bull 1991;27: 315-9.
Robert A. Rissman,Angel L. De Blas and David M. Armstrong(2007), GABAA receptors in aging and Alzheimer’s disease. Journal of Neurochemistry, 103, 1285–1292.
Shu-Chun Lin, Suu-Yi Lu, Szu-Ying Lee, Chi-Yen Lin, Chun-Hsien Chen and Kuo-Wei Chang( 2005).Areca (betel) nut extract activates mitogen-activated protein kinases and NF-jB in oral keratinocytes. Int. J. Cancer: 116, 526–535.
Smismans A, Schuit F, Pipeleers D. Nutrient regulation of gamma-
aminobutyric acid release from islet beta cells. Diabetologia 1997
;40:1411-5.
T.L. Kash, J.R. Trudell and N.L. Harrison(2004) Structural elements involved in activation of the γ -aminobutyric acid type A (GABAA) receptor. Biochemical Society Transactions Volume 32, part 3.
Trivedy C, Warnakulasuriya S, Peters TJ. Areca nuts can have
deleterious effects. Bmj 1999;318:1287.
Verspohl EJ, Tacke R, Mutschler E, Lambrecht G. Muscarinic
receptor subtypes in rat pancreatic islets: binding and functional
studies. Eur J Pharmacol 1990;178:303-11.
李啟賓:檳榔萃取液對雄鼠內生性抗氧化作用及生殖效應之研究，大
仁科技大學環境管理研究所.碩士論文(2008).
李音惠: 周邊GABA 對於雄鼠胃排空的抑制效應，國立陽明大學醫學院生理學研究所.碩士論文(2006)
鄭景暉. 嚼食檳榔的健康危害(不包括口腔癌與口腔癌前病變), 中華
民國92年.[[abstract]]近年來許多研究報告紛紛出爐，指出嚼食檳榔可能誘發其他多種系統疾病，包括氣喘(asthma) 、心律不整(cardiac arrhythmia) 、糖尿病(diabetes) 、免疫系統之影響、呼吸系統疾病、心血管系統、消化系統、神經系統功能、泌尿生殖系統之障礙等。嚼食檳榔對神經系統之影響已被確定，可在中樞神經系統與末梢神經系統而產生不同的影響，主要是因為檳榔子（Betel nut）中的檳榔素(Arecoline)與檳榔次鹼(Arecaidine)有關，這兩種檳榔生物鹼(alkaloids)可作用在毒蕈鹼受體(muscarinic receptor) 。 GABA為中樞神經系統中主要的抑制性神經傳導物質，目前已知有三種GABA 受體存在：GABAA、GABAB 與GABAC 受體，過去研究證實周邊神經與組織也有GABA與其受體的存在，GABA 是CNS 中最主要的抑制性神經傳導物質，參與約30% 的腦神經突觸的訊息傳遞，並且調節大部分神經的生理活動。本實驗目的以檳榔萃取液餵食實驗動物，藉由天數不同觀察雄鼠各個腦區其GABA Receptor的表現量，在實驗中我們使用（100 mg/kg/day）荖花檳榔劑量，以餵管方式分別餵食三組(15、30、45天)並犧牲取全腦，萃取mRNA並以RT-PCR方式反轉錄cDNA，再以Real – Time PCR分析其基因表現量。研究結果發現以檳榔萃取液餵食45天的雄鼠，其GABAA Receptor表現量明顯上升，顯示檳榔萃取液可促進老鼠腦部之GABAA Receptor表現量。另外，我們利用西方墨點法的技術來觀察是否GABAA Receptor的蛋白質表現量，經由ERK/MAPK pathway的調控，跟mRNA表現量一致。在未來我們將陸續進行一系列檳榔與神經系統的研究，獲得更多科學證據證明檳榔對於神經系統的傷害，並期盼未來研究能作為臨床醫學上之參考指標，藉此讓國人與衛生單位重視此現象，進而達到宣導與預防之目標。
ABSTRACT
It is estimated that 2 million person in Taiwan have the habit of chewing betel. Reports have shown that chronic betel quid (BQ) chewing is associated with diverse systemic effects, including , Diabetes Hypertension,Metabolic syndrome. There are four main arecal alkaloids, arecoline,arecaidine, guvacine and guvacoline. They also bind to GABA receptors in the brain, contributing to their psychoactive effects. Arecoline is not a simple activating ligand for the GABA receptor since it acts as a GABA receptor `blocker’, preventing normal GABA inhibition of neurotransmission. Previous studies show that chewing BQ may affect the nervous system.We investigated whether administration of Betel Quid Extraction (BQE) induces nervous transmission in the brain of SD rats. BQE (100mg/kg weight/day) was administered orally for 15, 30, 45 day.In addition, parameters studied inlucued the time course of mRNA expression of induced by BQE in SD rats.The results showed that GABRE gene mRNA expression to appear up-regulation. Mitogen - activated protein kinase superfamilies, including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinases (JNK) and p38, together with transcription factor GABRE, are important signaling elements. We examined the activation of these signaling pathways in SD Rat, treated with Betel Quid Extraction (BQE).In addition, recent reports have raised the potential links between BQ chewing and cancers of oropharynx, esophagus, stomach, urinary bladder and liver. Further in-depth studies are needed to highlight the association between BQ chewing and the body health
The Effect of Rhodopseudomonas sphaeroides to Nitrogen Source in Aquatic System
No full text英文摘要 ................................................ IV
致謝 .................................................... V
目錄 .................................................... VI
圖目錄 .................................................. XI
表目錄 .................................................. XIV
第壹章 緒論 ............................................. 1
1.1. 研究動機 ........................................... 1
1.2. 研究目的 ........................................... 3
第貳章 文獻回顧 ......................................... 5
2.1. 前言 ............................................... 5
2.2. 光合菌簡介與應用 ................................... 11
2.3. 光合反應的固氮和能量獲得的形式 ...................... 16
2.3.1. 光合反應的固氮方式 ............................... 16
2.3.2. 光合菌的獲能形式 ................................. 17
2.4. 光合菌的營養需求 ................................... 18
2.4.1. 光合菌的碳源 ..................................... 18
2.4.2. 光合菌的氮源 ..................................... 19
2.5. 影響固氮作用的因素 ................................. 20
2.6. 光合菌除氮功能研究現況 ............................. 21
2.6.1. 光合菌之研究現況 ................................. 21
2.6.2. 現今生物性除氮方式 ............................... 23
第參章 實驗材料與分析方法 ............................... 26
3.1. 實驗材料 .......................................... 26
3.1.1. 微生物 .......................................... 26
3.1.2. 本實驗使用之培養基 ............................... 27
3.1.3. 實驗用藥品 ....................................... 28
3.2. 實驗用儀器與設備 ................................... 32
3.2.1. 儀器設備 ......................................... 32
3.2.2. 光合菌培養條件和測試儀器 ........................... 32
3.3. 光合菌的培養方法 .................................... 33
3.3.1. 全封閉式厭氣光照培養(厭氣光合菌的培養) ............. 33
3.3.2. 光合菌菌種保存 .................................... 34
3.3.3. 光合菌解凍生長 .................................... 34
3.4. 分析方法 ............................................ 34
3.4.1. 細胞固定化 ........................................ 34
3.4.2. 分光光度計測量方法 ................................. 34
3.4.3. 血球計數盤 ....................................... 35
3.4.4. 水中銨分析 ........................................ 36
3.4.5. 水中亞硝酸鹽(NO2)的分析 ........................... 38
3.4.6. 格蘭氏染色法 ..................................... 39
第肆章 實驗設計 ......................................... 42
4.1. 預備試驗 ............................................ 42
4.1.1 採樣時間與地點 ..................................... 42
4.1.2 水樣採集 ........................................... 42
4.1.3 水質分析方法 ....................................... 42
4.2. 光合菌之培養、定性、定量 ............................. 43
4.2.1 光合菌之收集及吸收波長鑑定 .......................... 43
4.2.2 標準曲線製作 ....................................... 44
4.2.3 光合菌培養環境的設計與製作 ......................... 45
4.2.4 光合菌之生長曲線製作 ............................... 46
4.3. 不同濃度的氮源對光合菌之影響 ........................ 48
4.3.1 針對不同銨氮濃度對光合菌之影響 ......................48
4.3.2 針對不同亞硝酸鹽濃度對光合菌之影響 ................. 48
4.4. 不同菌量的光合菌對氮源之影響 ........................ 49
4.4.1 針對不同光合菌菌量對銨氮混合溶液影響 ................ 49
4.4.2 針對不同光合菌菌量對亞硝酸鹽混合溶液影響 ............ 49
4.5. 應用於養殖池水之模擬試驗 ............................. 50
4.6. 實驗流程 ............................................ 54
4.7. 除氮率計算公式 ...................................... 55
第伍章 結果與討論 ......................................... 56
5.1. 預備試驗 ............................................ 56
5.2. 光合菌之培養、定性、定量 .............................59
5.2.1 R. sphaeroides 吸收光譜 ............................ 59
5.2.2 R. sphaeroides 外型 ................................ 61
5.2.3 光合菌的培養環境 ................................... 62
5.2.4 光合菌之標準曲線製作 ............................... 65
5.2.5 光合菌之生長曲線製作 ................................ 66
5.2.6 銨氮濃度及亞硝酸鹽濃度之標準曲線製作 ................. 68
5.3. 不同濃度的氮源對光合菌之影響 ......................... 70
5.3.1 針對不同銨氮濃度對光合菌之影響 ...................... 70
5.3.2 針對不同亞硝酸鹽濃度對光合菌之影響 .................. 72
5.4. 不同菌量的光合菌對氮源之影響 ......................... 74
5.4.1 針對不同光合菌菌量對銨氮溶液影響 .................... 74
5.4.2 針對不同光合菌菌量對亞硝酸鹽影響 .................... 77
5.5. 應用於養殖池水之模擬試驗 ............................ 80
5.5.1 養殖池水對光合菌去除銨氮效果影響試驗 ............... 80
5.5.2 養殖池水對光合菌去除亞硝酸鹽效果影響試驗 ........... 82
5.5.3 養殖池水中含光合菌及不含光合菌的酸鹼度變化 ......... 84
第陸章 結論與建議 ....................................... 87
6.1. 結論 .............................................. 87
6.2. 建議 ............................................... 88
第柒章 參考文獻 ......................................... 90
圖目錄
圖2-1、全面的疾病防治系統 ............................... 6
圖2-2、氮循環 ........................................... 7
圖2-3、養殖池中的共生關係 ................................. 8
圖2-4、益生菌利用周遭產物淨化環境 ......................... 10
圖2- 5、Metabolic processes of R. palustris. ............. 22
圖2- 6、 Rhodopseudomonas sphaeroides(ESM) ............... 22
圖2-7、光合菌光合作用 .................................... 23
圖3-1、血球計數盤 .........................................36
圖3-2、格蘭氏染色劑 ....................................... 41
圖4-1、對半稀釋法 ......................................... 44
圖4-2、光合反應培養系統 ................................... 45
圖4-3、不再添加營養物質密閉空間之微生物生長曲線圖 .......... 47
圖4-4、實驗室內鰻魚養殖魚缸 ............................... 53
圖4-5、光合菌對氮源影響實驗設計流程圖 ..................... 55
圖5-1、嘉義鰻魚養殖池 .................................... 57
圖5-2、R. sphaeroides 吸收光譜 ........................... 60
圖5-3、使用結晶紫染色R. sphaeroide 放大400 倍 ............ 61
圖5-4、光合菌反應器 ...................................... 63
圖5-5、光合菌反應器內部照度值及溫度比較 ................... 64
圖5-6、R. sphaeroides 定量標準曲線 ....................... 65
圖5-7、R. sphaeroides 之生長曲線與pH 變化 ................ 67
圖5-8、R. sphaeroides 接種後培養第一小時 ................. 67
圖5-9、R. sphaeroides 接種後培養第九十六小時 .............. 68
圖5-10、銨氮濃度之標準曲線 ................................. 69
圖5-11、亞硝酸鹽濃度之標準曲線 .............................. 69
圖5-12、不同濃度的銨氮對光合菌之影響 ......................... 71
圖5-13、不同濃度的銨氮對光合菌之除氮率比較 ...................... 71
圖5-14、不同濃度的亞硝酸鹽對光合菌之影響 ....................... 73
圖5-15、不同濃度的亞硝酸鹽對光合菌之除氮率比較 ..................73
圖5-16、不同菌量的光合菌對銨氮溶液影響 ......................... 75
圖5-17、不同菌量的光合菌對銨氮之除氮率比較 ..................... 76
圖5-18、不同菌量的光合菌對亞硝酸鹽影響 ......................... 78
圖5-19、不同菌量的光合菌對亞硝酸鹽之除氮率比較 ................. 78
圖5-20、不同光合菌菌量對銨氮及亞硝酸鹽影響顏色變化圖 ............ 79
圖5-21、養殖池水中去除銨氮效果影響試驗 ......................... 81
圖5-22、養殖池水中去除亞硝酸鹽效果影響試驗 ..................... 83
圖5-23、養殖池水中除氮效率 .................................... 83
圖5-24、加入光合菌及沒有加入光合菌溶液的酸鹼度變化 ............. 86
圖5-25、養殖池水對光合菌除氮效果影響試驗其顏色變化 ............. 86
表目錄
表2-1、光合菌菌體組成 ........................................ 12
表2-2、光合菌B 群維生素組成 ................................... 12
表2-3、具有氫氣之生產能力之厭氧光合菌 ......................... 15
表2-4、光合菌的分類學特徵 .................................... 16
表2-5、在各種培養條件下光合菌的生長及其獲能形式 ............... 18
表2-6、A comparison of NH4+, NO2- ,DO,BOD, H2S,pH and Temp. . 57
表2-7、A comparison of NH4+, NO2- , PO4, ΣN, ΣP ............ 58
表2-8、現今生物性除氮歸納 .................................... 25
表3-1、BCRC13100 紫色不含硫菌Rhodopseudomonas sphaeroides..... 26
表3-2、Medium 66 ............................................ 27
表3-3、紫色不含硫菌的生長的培養配方 ........................... 27
表4-1、各種菌綠素的吸收波長 .................................. 44
表4-2、不同濃度的氮源對光合菌之影響不同銨氮及亞硝酸鹽濃度配置表 51
表4-3、不同菌量的光合菌對氮源之影響不同光合菌菌量配置表 ........ 52
表4-4、應用於養殖池水之模擬試驗養殖池水之菌量及無機氮配置表..... 53
表5-1、A comparison of NH4+, NO2- ,DO,BOD, H2S,pH and Temp.,℃.57
表5-2、A comparison of NH4+, NO2- , PO4, ΣN, ΣP ............ 58
表5-3、不同濃度的銨氮對光合菌之除氮率比較表 .................... 71
表5-4、不同濃度的亞硝酸鹽對光合菌之除氮率比較表 ................ 73
表5-5、不同菌量的光合菌對銨氮之除氮率比較表 .................... 76
表5-6、不同菌量的光合菌對亞硝酸鹽之除氮率比較表 ................ 79
表5-7、養殖池水中除氮效率 ..................................... 84
表5-8、光合菌於養殖池水中pH 值變化 ............................. 85
參考文獻
Aiking, H. and Sojka, G., 1979. Response of Rhodopseudomonas capsulata to illumination and growth rate in a light-limited continuous culture. J. Bacteriol., 139(1):530-536.
Armstong, D.A., 1979. Nitrogen toxicity to crustacea and aspects of its dynamics in culture system. In : D.Lewis and J. Ling (eds.), 2nd Biennial Crustacean Health Workshop. Texas A and M Sea Grant, TAMM-SE-79-114, Texas, pp. 329-360.
Austin,B.(1985) Antibiotics pollution from fish farm :effects on aquatic .Microbiol.Sci.,2(4):113-7.
Berger's Manual of Systematic Bacteriology, Vol. 1. Williams & Wilkins, Baltimore, MD, 1984, p.545-548.
Bovendeur, J., Eding, E.H. and Henken, A.M., 1987. Design and performance of a water recirculative system for high-density culture of the African catfish, Clarias gariepinus. Aquaculture, 63: 329-353.
Brunson, M.W., C.G. Lutz and R.M. Durborow. 1994. Algae blooms in commercial fish production farms. Southern Regional Aquaculture Center (SRAC) Publication No. 466.
Burrow, R.E., 1964. Effects of accumulated excretory products on hatchery-reared salmons. Bur. Sport. Fish. Wild. Res. Rep. No. 66.pp.12.
Cameron, J.N., 1971. Methemoglobin in erythrocytes of rainbow trout. Comp. Physical. 40A:743-749.
Campbell, J.W. 1973. Nitrogen excretion. In: C.L. Prosser (Ed) Comparative Animal Physiology. Third Edition W.D. Saunders, Toronoto, Ont: 966 pp.
Castillo F.,Caballero F.J.and Conrado M.,1986,“Nitrite uptake system in photosythetic bacteria Rhodopseudomonas capsulata E1F1.”Biochimica Biophysica Acta.848,16-23.
Chen, J.C. and Kou, Y.Z., 1992. Effects of ammonia on growth and molting of Penaeus japonicus juveniles. Aquaculture, 104: 249-260.
Chen, J.C., Liu, P.C. and Lei, S.C., 1990. Toxicities of ammonia and nitrite to Penaeus Monodon adolescents. Aquaculture. 89: 253-262.
Chiang, H.C. and Lee, J.C., 1986. Study of treatment and reuse of aquaculture wastewater in Taiwan. Aquacul. Eng., 5: 301-312.
Chien, Y. H. and Liang, R. Y.,1994,Paper presented in World Aquaculture ’94, New Orleans, Louisiana, Jan. p.12.
Chien, Y.H. and Wei, T.Z., 1993. The effects of photosynthetic bacteria Rhodopseudomonas capsulatus on submerged sediment chemistry under various temperature, salinity, and light intensity regime. Paper presented in Would Aquaculture ''93, Torremolinos, Spain, May 26-28.
Colt, J.E. and Tchobanoglous, G.T., 1978. Evaluation of short-term toxicity of nitrogenous compounds to channel catfish, Ictalurud punctatus. Aquaculture, 8: 209-224.
Dierstein, R. and Drews, G., 1975. Control of composition and activity of the photosynthetic apparatus of Rhodopseudomonas capsulata grown in ammonium-limited continuous culture. Arch. Microbiol., 106(3): 227-235.
Dobao, M.M. Luque, M.M. and Castillo, F., 1993. Nitrate reductase activity in spheroplasts from Rhodobacter capsulatus E1F1 requiresa periplasmic protein. Arch. Microbiol., 160: 471-476.
Herbert, R. A., 1976. Isolatio and identification of photosynthetic bacteria (Rhodospirillaceae) from antarctic marine and freshwater sediments. J. Appl. Bacteriol., 41: 75-80.
Hirotani, H., Ohigashi, H., Kobayashi, M., Koshimizu, K. and Takahashi, E. FEMS Microbiol. Lett. 77,p.13, 1991.
Nagadomi,H. Hiromitsu,T. Takeno,K. Watanabe,M and Sasak,K ,1999 ; Treatment of aquarium water by denitrifying photosynthetic bacteria using immobilized polyvinyl alcohol beads ; JOURNAL OF BIOSCIENCE AND BIOENGINEERING;Vol. 87, No. 2, 189-193.
Imhoff, J.F., 1982. Occurrence and evolutionary significance of two sulfate assimilation pathways in the Rhodospirillaceae. Arch. Microbiol. 132: 197-203.
Imhoff, J.F., Truper, H.G. and Pfenning, N., 1984. Rearrangement of the species and genera of the phototrophic purple nonsulfur bacteria. J. System. Bacteriol., 34(3): 340-343.
Kim J.S.,Ito K. and Takahashi H.,1981 “Production of molecular hydrogen by Rhodopseudomonas sp..” J. Ferment. Technol., Vol. 59,p.185-190.
Klaas J. Hellingwerf, WIM DE VRIJ, AND WIL N. KONINGS,Wavelength Dependence of Energy Transduction in Rhodopseudomonas sphaeroides: Action Spectrum of Growth,JOURNAL OF BACTERIOLOGY,Vol. 151,p. 534-541,Aug. 1982.
Kobayashi., M 1972. Utilization of photosynthetic bacteria. Proc. Ifs. Ferment. Technol., 527-531.
Koiller, M. and Avtalion, R.R., 1985. A laboratory scale recycling water unit for tilapia breeding. Aquacult. Eng., 4: 235-246.
Kranz R. G. and Haselkorn R.,1986, “Anaerobic regulation of nitrogen - fixation genes in Rhodopseudomonas capsulata. ”Proc.Natl.Acad.Sci.83,6805-6809.
Tseng,Kuo-Feng,2004, Submerged biofilter used in a recirculating eel culture system,Aquacultural Engineering 31,17-30.
Lascelles, J., 1959. Biochem. J. 72: 508.
Lee, C. M., Chen, P. C., Wang, C. C., Tung, Y. C., 2002 . Photohydrogen production using purple nonsulfur bacteria with hydrogen fermentation reactor effluent. International Journal of Hydrogen Energy, 27(11-12).
Lee.kwangyong, Nitrogen Removal from Wastewaters by Microalgae Without Consuming Organic Carbon Sources, J.microbiol.Biotechnol.,2002,12(6),979-985.
Lloyd, R. and Orr, L.D., 1969. The diuretic response of rainbow trout to sublethal concentrations of ammonia. Water Res., 3: 335-344.
Madigan, M.T. and Gest, h., 1979. Growth of the photosynthetic bacterium Rhodopseudomonas capsulata chemoautotrophically in darkness with H2 as the energy source. J. Bacteriol. 137(1): 524-530.
Mangum, C.P., Cilverthron, S.U., Harris, J.L., Towle, D.W. and Krall, A.R., 1976. The relationship between blood pH , ammonia excreyion and adaption to low salinity in the blue crab Callinectes sapidus. J. Exp. Zool., 195: 129-136.
McEwan, A. G.,Greenfield A. J.,Wetzstein H.G.,Jackson J.B.and Ferguson S.J.,1985, “Nitrous oxide reduction by members of the family Rhodospirillaceae and the nitrous oxide reductase of Rhodopseudomonas capsulata. ”J.bacteriol.164,823-830.
Miller, G.E. and Libey, G.S., 1984. Evalution of a trickling in a recirculating aquaculture system containing channel catfish. Aquacult. Eng., 3:39-57.
Olson, K.R. and Fromm, P.O., 1971. Excretion of urea by two teleost exposed to different concentration of ambient ammonia. Comp. Biochem. Physiol., 40A: 999-1007.
Pfennig, N. 1967. Annu. Rev. Microbiol., 21, 285.
Pfennig, N. Ann. Rev. Microbiol. 31, p.275, 1977.
Pillay ,T.V.R.,1992.Aquaculture and Environment. Halsted Press. New York ,Toronto.
Poxton, M.G. and Allouse, S.B., 1987. Cyclical fluctuation in ammonia and nitrite-nitrogen resulting from the feeding of turbot, Scophthalmus maximus, in recirculating system. Aquacult. Eng., 6: 301-322.
Reddy, K.R. and Patrick, Jr. W. H., 1984. Nitrogen transformation loss in flooded soils and sediment. CRC Critical Review in Environ. Control., 13: 273-309.
Sasikala, C. and Ramana, C. V., Adv. Appl. Microbiol. 41,p. 174,1995.
Sawada, H., Parr, R.C. and Rogers, P.L., 1977. Photosynthetic bacteria in waste treatment. J. Ferment. Technol., 55(4): 326-336.
Sharma, B. and R.C. 1977. Nitrification and nitrogen removal. Water Res., 11:879-925.
Smith, C.E. and W.G. Williams 1974. Experimental nitrite toxicity in rainbow trout and chinook salmon. Trans. Amm. Fish. Soc., 103:389-390.
Smith, C.E.and R.C. Russo 1975. Nitrite-induced methemoglobinemia in rainbow trout. Prog. Fish-Cult., 37(3):150-152.
Spotte, S. 1979. Fish and Invertebrate Culture, Water Management in Closed Systems, 2nd end, Wiley-Interscience, New York, 114 p.
Stevens, p., Sypt,H.V., Vos, P.D. and Ley, J.D., 1983. Comparative study on H2 evolution from DL-lactate, acetate and butyrate by different strains of Rhodopseudomonas capsulata in a new type of reactor. Bioterchnol. Lett., 5(6): 369-374.
Suhaimi, M., Kiessens, J. and Verstraete, W., 1987. NH4+-N assimilation by Rhodobacter capsulatus ATCC 23782 grown axenically and non-axenically in N and C rich media. J. Appl. Bacteriol., 62: 53-64.
Kikuta and Fukada, Water quality control of aquarium water for admired fishes. In: M. Murakami, Editor, Examples of purification of the waters for environmental use, Power-Sha, Tokyo (1996), pp. 11–17.
Takakuwa, S., Odom, J.M. and Wall, J.D., 1983. Hydrogen uptake deficient mutants of Rhodopseudomonas capsulatus, Arch. Microbiol., 136: 20-25.
Takayashi, M., and Ichimora, S., 1970. Photosynthetic properties and growth of photosynthetic bacteria in lakes. Limnol. Oceanogr., 15: 929-944.
Thurston, R.V., Russo, R.C. and Emerson, K, 1979. Aqueous ammonia equilibrium-tabulation of percent un-ionzed ammonia. Enviromental Protection Agency Ecological Research series EPA-600/3-79-091.
Vincenzini, M., Materassi,R., Sili,C., and Balloni, W., Int. J.Hydrogen Energy, 11, p.623, 1986.
Weston,D.P.(1996) Environmental considerations in the use of antibacterial drugs in aquaculture. In:Baird,D., nBeveridge ,M.V.M., Kelly,L.A.,Muir,J.F.(Eds.),Aquaculture and Water Resource Management.Blackwell,Oxford,140-165.
Willison, J. C., 1993. Biochemical genetics revisted: the use of mutants to stuy carbon and nitrogen metabolism in the photosynthetic bacteria. FEMS Microbiol. Rev., 104: 1-38.
Yen, H.C. and Marrs, B., 1977. Growth of Rhodopseudomonas capsulatus under anaerobic dark conditions with dimethyl sulfoxide. Arch. Biochem. Biophysiol., 181: 411-418.
小林正泰，1978，光合细菌处理高浓度有机废水《发酵与工业(日)》，1978 / 36 / 09，P.753-760。
小林達治，1967，光合菌的大量培養方法，＜特許公報＞(日)，昭42-11979，1967。
小林達治，1975。光合成細菌 基礎 應用II。土肥誌，46，p.148-156。
冉繁華、陳秀男，2006，益生菌在養殖管理系統的研發與應用，縵魚養殖之健康管理，行政院農委會水產試驗所，167-190頁。
矢木修身，用水的廢水，（發酵的工業），19(8)，1977，949-951。
佐藤敏生，1984。光合成細菌 窒素代謝p. 47-61.北村 博、森田茂廣、山下仁平編，光合成細菌， 學會出版。
宋作人，1984，活性污泥法添加及未添加光合菌處理魚巿場廢水，國立成功大學環境工程研究所碩士論文。
林智中，光合菌的分離、培養及在養殖方面應用之研究，台灣國立海洋科技大學，碩士論文，1999，NO.088NTOU0086021。
張志強，2000，光合菌在光合菌反應器產氫之研究，國立中央大學化學工程研究所碩士論文。
張怡塘，2002，微生物實驗，高立出版，第二版修訂。
陳秀男 ，1994， 蝦病管理對策。行政院農委會，144 pp。
陳耕鎔，2007，光合菌對水中硝酸鹽去除率之研究 ，崑山環工所 ，專題研究。
黃世鈴，2006， 縵魚養殖之健康管理。行政院農委會水產試驗所。
黃汝賢、曾國鋒、廖一久，1991，養殖池水循環處理利用系統之探討，p.119~131。第十六屆廢水處理技術研討會論文集。
劉敏主，2006，血液學檢查法，行政院農業委員會家畜衛生試驗所參考手冊，動物傳染病實驗室檢驗方法第十章。http://vettech.nvri.gov.tw/Articles/handbook/73.html
澤田英子、佐藤敏生，1984，脫窒光合成細菌 脫窒素作用。P.175-180。北村 博、森田茂廣、山下仁平編，光合成細菌，學會出版 。
環境保護署環境檢驗所，2002，水質檢測方法，NIEA W418.51C。http://www.niea.gov.tw/niea/WATER/W41851C.htm
環境保護署環境檢驗所，2005，水質檢測方法，NIEA W446.52C。http://www.niea.gov.tw/niea/WATER/W446.52C.htm
魏志榮，陳瑤湖，1989。在不同溫度、鹽度、與光照度下光合菌對養殖池底質及水質之影響。國立海洋大學，水產養殖研究所。
羅美棧，孫全杰，羅玉輝，巫健次，1973。以光合菌及綠藻處理水肥之研究(第三期研究報告)。臺灣環境衛生，5(1)：21-39。
蘇偉成、劉富光，2005臺灣水產養殖的永續經營。科學發展，385期，42-49頁。[[abstract]]水中氮循環對水產養殖系統有極大的影響，氮源對於魚蝦貝類所產生的毒性傷害，往往是水產養殖系統中無形的殺手，而氮循環中固氮作用所產生的銨氮(Ammonium-NH4+)及亞硝酸鹽(Nitrogen dioxide-NO2- )，在海水或淡水中濃度高於2mg/L時，魚類即可能產生病變，有鑑於此，本研究期望藉由光合菌 (Photosynthesis Bacteria；PSB)有效控制水中無機氮之濃度，將對水產養殖業有所幫助。
本研究利用紫色不含硫光合菌(Rhodopseudomonas sphaeroides P4株菌)為材料，藉由此微生物製劑方式來控制水中氮源之含量，並對光合菌量的添加菌量及除氮效率之影響加以研究。
本實驗結果顯示光合菌培養至一百二十小時後，菌量可達飽和量濃度109no./ml，光合菌培養過程中的水中酸鹼值於第二十四小後開始趨向弱鹼性，其光合菌數與Abs(OD660)吸光值兩者的相關性R2=0.9941，我們將藉由Abs(OD660)所測得之吸光值作為定量光合菌濃度(Klaas,1982)。第二部分顯示，在五種不同水中亞硝酸鹽濃度下，菌量108 no. /ml於24小時後，水中亞硝酸鹽去除率達100%，另外光合菌在五種不同銨氮濃度下，光合菌菌量於108 no./ml作為120小時後，銨氮去除率平均都可達58%。第三部分顯示不同菌量濃度中，以109no. /ml菌量去除亞硝酸鹽效率最佳，而菌量108no. /ml去除銨氮效率最佳可逹93%。第四部分結果顯示，養殖池水對除氮效率上並無影響效果，但光合菌可穩定養殖池水中的酸鹼度。
The nitrogen cycle of aquatic ecosystem plays an important role, especially inaquatic farms. The effluent of nitrogen source is an invisible killer that could harm the aqua products. In the seawater or fresh water systems, the concentration of Ammonium(NH4+) and Nitrous acid(NO2-) higher than 2mg/L may cause the pathological change of fish. Therefore,this research was designed to study the use of Photosynthetic bacteria (PSB) to biomanipulate the concentration of inorganic nitrogen in the aquatic system. It might be helpful to the aquatic farms and aquatic environments if the increase of nitrogen concentrate is controlled efficiently. Rhodopseudomonas sphaeroides was used in this study and Microbial Pesticides methods were applied to control the content of nitrogen source in aquatic experiments.
The results show that the amount of bacteria was saturated after 120 hours culture of R. sphaeroides. The saturated bacterial concentration reached to 109no./ml. In the process of culture, the pH value becomes weak alkalescence after 24 hour culture. The correlation between the amount of R.sphaeroides and the absorbance value of optical density at 660 nm is R2=0.9941. The Abs(OD660) values were utilized to measure the quantities of R. sphaeroides(Klaas,1982). Then, R. sphaeroides (108 no./ml) were added in five different concentrations of Nitrous acid in water to study the removal of Nitrous acid after 24 hours. The results show that all Nitrogen acid can be removed by R. sphaeroides completely. Next, R. sphaeroides (108 no./ml) were added in five different concentrations of Ammonium to study the removal rate of Ammonium. The removal rate of Ammonium is about 58% after 120 hours. Third, among three levels of bacterium concentration, the concentration of 109no. /ml is the best for removing Nitrous acid. Fourth, R. sphaeroides can stabilize the pH value in the water of aquaculture pond, but it has no effect on the removal of Nitrogen
Hyperosmolarity enhanced the fibrosis susceptibility by phosphorylation of type II TGF-β receptors and up-regulation of Tbeta-RI in MDCK cells
No full text中文摘要 i
英文摘要 iii
致謝 v
縮寫表 vii
第一章 緒論
一、糖尿病與糖尿病腎病變之盛行率 1
二、甘露醇（D -mannitol）簡介 2
三、腎纖維化之致病機轉 3
四、乙型轉型因子以及乙型轉型因子受器結構與命名 4
五、乙型轉型生長因子之作用與調節 6
六、腎纖維化與乙型轉型生長因子受器之作用 7
七、纖維化與乙型轉型生長因子之訊息傳導路徑 8
八、蛋白質降解之泛素蛋白酶體降解途徑 12
九、研究目的 14
第二章 材料與方法
一、試劑與材料 15
二、方法 16
1. 細胞培養 16
(1) 繼代細胞 16
(2) 活化細胞 17
(3) 凍細胞 17
2. 細胞數目 17
3. 細胞存活率 18
4. 酵素連結免疫吸附分析(ELISA) 18
5. 總量核酸核酸萃取及cDNA的製備 18
6. 反轉錄聚合酵素連鎖反應(RT-PCR) 19
7. 瓊脂膠體電泳 20
8. 細胞內第二型乙型轉型生長因子受器磷酸化分析 20
9. 細胞蛋白質總量測定 21
10.西方墨點分析 21
11.免疫沉澱法 23
12.統計分析 24
第三章 結果
一、高滲透壓以及乙型轉型生長因子對纖維蛋白之影響 25
二、高滲透壓對乙型轉型生長因子之影響 26
三、高滲透壓對乙型轉型生長因子接受器之影響 26
1、高滲透壓對乙型轉型生長因子受器蛋白質表現之影響 26
2、高滲透壓對第二型乙型轉型生長因子受器磷酸化之影響 27
四、高滲透壓增加第一型乙型轉型生長因子受器蛋白質之探討
27
五、高滲透壓對乙型轉型生長因子受器蛋白質上Smad 7- Smurf l
之影響 28
六、高滲透壓導致第一型乙型轉型生長因子受器蛋白質半衰期增加
29
七、高滲透壓對乙型轉型生長因子訊息傳導路徑-Smad途徑之影響
30
第四章 討論 31-36
第五章 圖表 36-45
第六章 參考文獻 46-53
參考文獻
Ateshian, G. A., Soltz, M. A., Mauck, R. L., Basalo, I. M., Hung, C. T. & Lai, W. M. 2003 The role of osmotic pressure and tension-compression nonlinearity in the frictional response of articular cartilage. Transport in Porous Media 50, 5–33.
Attisano L, Wrana JL, Lopez-Casillas F, Massague J: TGF-beta receptors and actions. Biochem Biophys Acta 1222:71-80, 1994
Ayo SH, Radnik R, Garoni JA, Troyer DA, Kreisberg JI: High glucose increase DAG mass and activates PKC in mesangial cell cultures. Am J Physiol 261:F571-577,1991
Border WA, Okuda S, Languino LR, Ruoslahti E: Transforming growth factor-b regulates production of proteoglycans by mesangial cells. Kidney Int 37:689-695, 1990
Border WA, Noble NA. Transforming growth factor beta in tissue fibrosis. N Engl J Med 1994; 331: 1286–92.
Border WA, Noble NA. Evidence that TGF- should be a therapeutic target in diabetic nephropathy. Kidney Int 1998; 54: 1390–1391.
Ceshi Chen & Lydia E. Matesic. The Nedd4-like family of E3 ubiquitin ligases and cancer. Cancer Metastasis Rev (2007) 26:587–604DOI 10.1007/s10555-007-9091-x
Chen Y, Lebrun JJ, Vale W. Regulation of transforming growth factor - and activin-induced transcription by mammalian Mad proteins. Proc Natl Acad Sci USA 93, 12992-12997, 1996
Chiasson JL, Aris-Jilwan N, Belanger R, et al. Diagnosis and treatment of diabetic ketoacidosis and hyperglycemic hyperosmolar state. CMAJ 2003; 168: 859-66
Ciehanover A, Hod Y, Hershko A. (1978). A heat-stable polypeptide component of an ATP-dependent proteolytic system from reticulocytes. Biochem Biophys Res Commun. 81(4):1100–1105.
Ciechanover A. (2000). Early work on the ubiquitin proteasome system, an interview with Aaron Ciechanover. Cell Death Differ Sep;12(9):1167–77.
de Larco, J.E. and Todaro, G.J. (1978) Growth factors from murine sarcoma virus-transformed cells. Proc Natl Acad Sci U S A, 75, 4001-4005.
Derynck R: TGF-beta-receptor-mediated signaling. Trends Biochem Sci 19:548-553, 1994
Ebner R,Chen RH,Shum L,et al. Cloning of a type I TGF-beta receptor and its effect on TGF-beta binding to the type Ⅱ receptor Science,1993,260(5112):1344.
Ebner R, Chen RH, Lawler S, Zioncheck T, Derynck R. Science 262,900-902, 1993
Eddy AA: Molecular basis of renal fibrosis. Pediatr Nephrol 15: 290–301, 2000.
Goldknopf IL, Busch H. (1977). Isopeptide linkage between nonhistone and histone 2A polypeptides of chromosomal conjugate-protein A24. Proc Natl Acad Sci USA 74(3):864-8.
Gudrun Valdimarsdottir, Marie-José Goumans, Fumiko Itoh, Susumu Itoh3, Carl-Henrik Heldin3 and Peter ten Dijke. Smad7 and protein phosphatase 1α are critical determinants in theduration of TGF-β/ALK1 signaling in endothelial cells.BMC Cell Biology 2006, 7:16 doi:10.1186/1471-2121-7-16
Haas AL, Warms JV, Hershko A, Rose IA. Ubiquitin-activating enzyme: Mechanism and role in protein-ubiquitin conjugation. J Biol Chem. 1982 March 10;257(5):2543–8.
Heldin, C.H., Miyazono, K. and ten Dijke, P. (1997) TGF-beta signalling from cell membrane to nucleus through SMAD proteins. Nature, 390, 465-471.
Hostetter TH: Diabetic nephropathy. In: Brenner BM, Rector RC eds. The kidney. pp.1695-1727. W.B. Saunders Co. Philadelphia, USA, 1991.
Hung-Chang Liu, Tung-Nan Liao, Tao-Chen Lee, Fen Liu, Lea-Yea, Chuang, Jinn-Yuh GuhShu- Min-Shou Hu, Ya-Lin Yang, Shyh-Horng Lin,Min-Yuan Hung, Jau-Shyang Huang, Tsung-Jen Hung, Chun-Da Chen,Tai-An Chiang, Jun-Yu Chan,Sen-Yi Chen, and Yu-Lin Yang. Albumin Induces Cellular Fibrosis by Upregulating Transforming Growth Factor-Beta Ligand and Its Receptors in Renal Distal Tubule Cells. Grant sponsor: National Science Council, Taiwan; Grant number: NSC93-2314-B-273-001.
Husnjak K, Elsasser S, Zhang N, Chen X, Randles L, Shi Y, Hofmann K, Walters KJ, Finley D, Dikic I. (2008). Proteasome subunit Rpn13 is a novel ubiquitin receptor. Nature 22, 481–488.
Itoh S, Landstrom M, Hermansson A, Itoh F, Heldin CH, Heldin NE & ten Dijke P 1998 Transforming growth factor beta1 induces nuclear export of inhibitory Smad7. Journal of Biological Chemistry 273 29195–29201.
J Hand Surg [Br]. (2006) Feedback inhibition of high TGF-beta1 concentrations on myofibroblast induction and contraction by Dupuytren's fibroblasts. Epub Oct;31(5):473-83.
Kitabchi AE, Umpierrez GE, Murphy MB, et al. Management of hyperglycemic crises in patients with diabetes. Diabetes Care 2001; 24: 131-53.
Kretzschmar M, Liu F, Hata A, Doody J, Massague J: The TGF-b family mediate Smad1 is phosphorylated directly and activated functionally by the BMP receptor kinase. Gene Dev 11,984-995, 1997
Lai, W. M., Hou, J. S. & Mow, V. C. 1991 A triphasic theory for the swelling and deformation behaviours of articular cartilage. J. Biomech. Eng. 113, 245–258.
Lawler S: The type II transforming growth factor-b receptor autophosphorylates not only on serine and threonine but also on tyrosine residues. J Biol Chem 272, 14850-14859, 1997
Leask, A. and Abraham, D. J. (2004). TGF- signaling and the fibrotic response. FASEBJ. 18, 816-827
Lodish, H, Berk A, Matsudaira P, Kaiser CA, Krieger M, Scott MP, Zipursky SL, Darnell J. (2004). Molecular Cell Biology, 5th ed., ch.3, pp 66–72. New York: WH Freeman.
LeRoy EC, Trojanowska MI, Smith EA. Cytokines and human fibrosis. Eur Cytokine Netw 1990; 1: 215-219
Massague´, J. (1998) Annu. Rev. Biochem. 67, 753–791
Massague J. Receptors for the TGF-beta family. Cell,1992,69(7):1067.
Milhorat TH, Pan J. The blood-brain barrier and cerebral edema. In: Cottrell JE, Smith DS, eds. Anesthesia and Neurosurgery. St. Louis: Mosby; 2001:115-127.
Nakamura T: Isolation and characterization of activin receptor from mouse embryonal carcinoma cells: Identification of its serine/threonine/tyrosine protein kinase activity. J Biol Chem 267, 18924-18928, 1992
Nakao A: TGF- receptor-mediated signaling through Smad2, Smad3 and Smad4. EMBO 16, 5353-5362, 1997
Nakao A: Identification of Smad7, a TGF- inducible antagonist of TGF- signaling. Nature 389, 631-635, 1997
Nakao A, Afrakhte M, Moren A, Nakayama T, Christian JL, Heuchel R, Itoh S, Kawabata M, Heldin NE, Heldin CH et al. 1997 Identification of Smad7, a TGFbeta-inducible antagonist of TGF-beta signalling. Nature 389 631–635.
Rik Derynck & Ying E. Zhang: Smad-dependent and Smad-independent pathways in TGF- family signalling Nature 425, 577-584 (9 October 2003)
Risseeuw EP, Daskalchuk TE, Banks TW, Liu E, Cotelesage J, Hellmann H, Estelle M, Somers DE, Crosby WL. Protein interaction analysis of SCF ubiquitin E3 ligase subunits from Arabidopsis. Plant J. 2003 Jun;34(6):753-67.
Rocco MV, Chen Y, Goldfarb S: Elevated glucose stimulates TGF- gene expression and bioactivity in proximal tubule. Kidney Int 41:107-114, 1992
Russette M,Lyons RM,Mosos HL. Transforming growth faclors and the regulation of cell proliferation. Eur J Biochem,1990,187(3):467.
Schiller M, Javelaud D, Mauviel A. TGF-beta-induced SMAD signaling and gene regulation: consequences for extracellular matrix remodeling and wound healing. J Dermatol Sci 2004; 35: 83-92
Schreiner P, Chen X, Husnjak K, Randles L, Zhang N, Elsasser S, Finley D, Dikic I, Walters KJ, Groll M. (2008). Ubiquitin docking at the proteasome through a novel pleckstrin-homology domain interaction. Nature 22, 548–552.
Sharma K, Ziyadeh FN, Alzahabi B et al. Increased renal production of
transforming growth factor-1 in patients with type II diabetes. Diabetes
1997; 46: 854–859.
Sharma K, Ziyadeh FN: The emerging role of transforming growth factor-b in kidney diseases. Am J Physiol 266: F829 –F842, 1994
Shenkin HA, Goluboff B, Haft H. The use of mannitol for reduction of intracranial pressure in intracranial surgery. J Neurosurg 1962; 19: 897-900.
Striker GE, Peten EP, Caromen MA, Pesce CM, Schmidt K, Yang C-W, Elliot SJ, Striker LJ: The kidney disease of diabetes mellitus (KDDM): a cell and molecular biology approach. Diabetes/Metab Rev 9: 37-56, 1993.
T Yamaguchi, A Kurisaki, N Yamakawa1, K Minakuchi2 and H Sugino: FKBP12 functions as an adaptor of the Smad7–Smurf1 complex on activin type I receptor. Journal of Molecular Endocrinology (2006) 36, 569–579
ten Dijke, P., and Hill, C. (2004) Trends Biochem. Sci. 29, 265–273
Takahito Itoh , Atsushi Yamauchi , Enyu Imai , Naohiko Ueda , Takenobu K a m a d a.(1995) . Phosphatase toward MAP kinase is regulated by osmolarity in Madin-Darby canine kidney (MDCK) cells. FEBS 123-126.
Takanori Ebisawa, Minoru Fukuchi, Gyo Murakami, Tomoki Chiba, Keiji Tanaka,Takeshi Imamura, and Kohei Miyazono.(2001) Smurf1 Interacts with Transforming Growth Factor- Type I Receptor through Smad7 and Induces Receptor Degradation. JOURNAL OF BIOLOGICAL CHEMISTRY, 12477–12480.
Thrower JS, Hoffman L, Rechsteiner M, Pickart CM. (2000). Recognition of the polyubiquitin proteolytic signal. EMBO J 19, 94–102.
Topper JN: Vascular MADs: Two novel MAD-related genes selectively inducible by flow in human endothelium. Proc Natl Acad Sci USA 94, 9314-9319, 1997
Uitto J, Kouba D. Cytokine modulation of extracellular matrix gene expression: relevance to fibrotic skin diseases. J Dermatol Sci 2000; 24 Suppl 1: S60-S69.
Verrecchia, F., Pessah, M., Atfi, A. and Mauviel, A. (2000) Tumor necrosis factor-alpha inhibits transforming growth factor-beta /Smad signaling in human dermal fibroblasts via AP-1 activation. J Biol Chem, 275,30226-30231.
Verrecchia F, Mauviel A. TGF-beta and TNF-alpha: antagonistic cytokines controlling type I collagen gene expression. Cell Signal 2004; 16: 873-880.
Wakefield LM, Winokur TS, Hollands RS,et al. Recombinant latent transforming growth factor beta 1 has a longer plasma half-life in rats than active transforming growth factor beta 1 and a different tissue distribution. J Clin Invest,1990,86(6):1976.
Wieser R, Wrana JL, Massague J: GS domain mutations that constitutively activate Tb R-I, the downstream signaling component in the TGF-b receptor complex. EMBO 14, 2199-2208, 1995
Wise BL, Chater N. The value of hypertonic mannitol solution in decreasing brain mass and lowering cerebrospinal fluid pressure. J Neurosurg 1962; 19:1038-43.
Wolf G, Sharma K, Chen Y: High glucose-induced proliferation in mesangial cells is reversed by autocrine TGF-beta. Kidney Int 42:647-656, 1992
Wrana JL (1998) TGF-beta receptors and signalling mechanisms. Miner Electrolyte Metab 24:120–130.
Yoshio Terada, Kimio Tomita, MiwakKo. HommaO, Hiroshi Nonoguchi, TiaxinY ang,Takehisa Yamada, Yasuhito YuasaO, Edwin G. Krebsn, Sei Sasaki, and FumiakMi arumo. Sequential Activation of Raf-1 Kinase, Mitogen-activated Protein( M A P ) Kinase Kinase, MAP Kinase, and S6 Kinase byHyperosmolality in Renal Cell. Journal of Biological Chemistry pp. 31296-31301, 1994
Young BA, Johnson RJ, Alpers CE, Eng E, Gordon K, Floege J, Couser WG, Seidel K. Cellular events in the evolution of experimental diabetic nephropathy. Kidney Int. 1995 Mar;47(3):935-44.
Zhang Y, Feng X, Wu R, Derynck R. Nature 383, 168-172, 1996
Zhang Y, Feng XH, Wu RY, Derynck R: Receptor-associated Mad homologues synergize as effectors of the TGF-beta response. Nature (in press), 1997
Ziyadeh FN, Snipes ER, Watanabe M: High glucose induces cell hypertrophy and stimulates collagen gene transcription in proximal tubule. Am J Physiol 259: F704-F714, 1990
Ziyadeh FN, Goldfarb S. The renal tubulointerstitium in diabetes mellitus. Kidney Int 1991;39:464–75.
Ziyadeh FN: Significance of tubulointerstitial changes in diabetic renal disease. Kidney Int 54(suppl): S10-13, 1996.
楊景民、郝麗瑾 (1995) 滲透壓及其臨床應用。荷澤醫專學報，7 (1)：3-6。[[abstract]]根據衛生署2006年所公佈的資料顯示，糖尿病為95年國人十大死因的第四名。高血糖高滲透壓狀態 (簡稱HHS) 是糖尿病最嚴重的急性併發症之一，其死亡率居高不下。
乙型轉型生長因子(TGF-β)是一個纖維激素，與腎臟纖維化的進展有相關，而隨後活化乙型轉型生長因子受器(TGF-βRII和TGF-βRI)，造成細胞外基質蛋白堆積，導致腎臟的纖維化。在這研究當中，我們利用甘露醇誘發高滲透壓來證明在遠端腎小管細胞中第二型乙型轉型生長因子受器磷酸化和第一型乙型轉型生長因子受器表現。我們使用狗腎遠端小管(MDCK)細胞，將細胞培養於不同滲透壓濃度的情況。
在高滲透壓下，經過乙型轉型生長因子(10ng/ml)之處理，藉由酵素連結免疫吸附分析來觀察纖維蛋白表現量。使用西方墨點法來研究乙型轉型生長因子受器以及下游蛋白質的表現。我們發現在甘露醇會提升乙型轉型生長因子所誘導纖維蛋白增加，然而，在滲透壓濃度劑量及時間點下，遠端小管細胞第二型乙型轉型生長因子受器上酪胺酸336位置磷酸化。高滲透壓下也會導致第一型乙型轉型生長因子受器以及乙型轉型生長因子下游訊息路徑Smad的表現。另外，使用cyclohexamide刺激，我們發現高滲透壓增加第一型乙型轉型生長因子受器半衰期，並且使抑制第一型乙型轉型生長因子受器走向 ubiquitination – proteosomal水解路徑。由以上這些結果來看，遠端腎小管細胞在高滲透壓會提升纖維化的敏感性，藉由第二型乙型轉型生長因子受器磷酸化以及第一型乙型轉型生長因子受器表現。
Diabetes mellitus have taken the fourth place in the ten major causes of the death of according to the reports issued by the Department of Health in Taiwan, 2006. Hyperglycemic hyperosmolar state (HHS) is one of the most serious acute complications of diabetes and its mortality rate had been higher.
Transforming growth factor beta 1(TGF-β1) is a fibrokine implicated in the progression of renal fibrosis, followed TGF-β1 receptor (TGF-β RII and TGF-β RI) activation resulting in accumulation of extracllular matrix protein and caused renal cellular fibrosis. In this study, we utilized mannitol-inducing hyperosmolarity to elucidate the phosphorylation of type II TGF-β receptors and expression of Tbeta-RI in distal renal tubular cells. We were used Madin-Darby Canine Kidney (MDCK) cells, cultured in the presence of various on concentration of osmotic.
Hyperosmolarity induce fibronectin protein level by TGF-β1(10ng/ml) using enzyme-linked immunosorbent assay. Western blotting were used to investigate the protein expression of TGF-βreceptors and downstream protein. We showed that mannitol enhanced TGF-β–induced increase in fibronectin in MDCK cells. Moreover, Osmotic concentration time- and dose- dependently induced the phosphorylation of Tbeta RII in tyrosine 336 in MDCK cells. Hyperosmolarity also induced increase expression of Tbeta RI and Smad signal pathway. In addition, by using cyclohexamide, we showed that hyperosmolarity significantly increased down-regulated the half-life and inhibited protein levels of TbetaRI by poly-ubiquitination and proteosomal degradation. These results suggest that hyperosmolarity increased the fibrosis susceptibility by phosphorylation of type II TGF-β receptors and up-regulation of the Tbeta-RI expression in MDCK cells
Recovery of Valuable Metals in Waste Batteries by a New Thermal Separation Method
No full text[[abstract]]新型高溫分離法回收電池中有價值金屬之研究隨經濟的快速發達，乾電池用量亦隨之逐年增加，近年來廢乾電池之處理廣泛受到關注，為此申請人以熱熔融法為原思考出發點，改良成新型之高溫分離技術來處理廢乾電池，本研究之目的係針對廢乾電池中有價值金屬之回收技術做整體探討，研究工作預計分成三年完成，第一年擬於實驗室中之高溫分離系統完成碳鋅電池之分離處理，希望能高效率回收電池中之Fe、Mn 與Zn，並釐清添加劑對分離機制之影響。第二年將修改系統以配合鎳鎘電池之特性，以高效率地回收電池中之Cd、Fe 與Ni，之後將整合前兩年實驗經驗，希望能在第三年於模廠進行實驗，分別測試碳鋅、鹼性與鎳鎘電池，以模廠操作經驗來研擬出渣系統之改善方法，並提出未來實廠運作時可行之設計方式，總體而言，本計畫將以提供關於高溫分離技術許多科學與實務上之資訊，提供從事相關領域研究之先進做參考。
With rapid economic developing, the amount of used batteries also increases significantly. The final disposal of used batteries is of concern at present so we apply a thermal separation technology of modified vitrification. The object of the three-year research is to evaluate the effect of this technology on the recovery of useful metals from used batteries. In this study, it is attempted to treat C-Zn batteries to recover C, Fe, Mn, and Zn with high efficiency and investigate the effect of the additives on the metal separation. In the second year, we will modify the thermal separation system for the treatment of Ni-Cd batteries and try to recover Cd, Fe and Ni effectively. Based on the operation experience during the first two years, it is planed to conduct the tests in a pilot plant. Three types of batteries, including C-Zn, alkaline, and Ni-Cd batteries, will be treated with different parameters. We wish to improve the way of discharging molten materials and provide useful experience for the operation in a real scale plant. Overall speaking, the whole project will provide much useful scientific and practical information for researchers who interest in the thermal separation technology
An Externsion of Trust and Privacy with TPB and TAM Model in the Intial Adoption of On-Line Shopping
No full text研究領域:管理科
Gene Therapy and Analysis of the Effect of Anti-Inflammatory Factors for Osteoarthritis
No full text研究領域:臨床醫學類, 生物技術
計畫編號:NSC98-2314-B273-001-MY
Squid Frozen Storage Quality Improvement and Product Development Research
No full text研究領域:食品科技, 漁業類
計畫編號:98農科-3.1.4-漁-F1(1)[[abstract]]1.計畫目標： (1)現有遠洋魷魚儲運品質易因魚獲後處理及冷凍條件之不穩定，造成魷魚外皮膜色素胞之變色，影響魷魚外觀及商品價值。為維持捕獲魷魚遠洋儲運中之品質，需提供業者可行的儲運品質提昇方案，確保長時低溫儲運過程中魚體表面乾燥、氧化及色素胞生理能機崩解造成的劣變現象得以控制並維持較高的商品價值。 (2)魷魚提供國人優良的蛋白質來源及豐富的高度不飽和脂肪酸，是國人良好的海洋食品來源。但魷魚肌肉蛋白質組織結構的特性，造成魷魚經過加熱或乾燥後質感大幅改變，形成硬而較具彈性的肉質組織，對幼兒及銀髮族而言並不是一種容易入口消化的蛋白質，為有效提昇資源的利用實需提供轉變原有肉質結構為易被消化分解的蛋白質的加工方式，並提高其附加價值，從單純的食品提昇為具保健機能特質的食品。產業結構層次藉此得以優質化及精緻化，創造更多的市場競爭力及經濟力。 2.重要工作項目： (1)針對魷魚遠洋儲運過程中魚體表皮因儲運環境產生的生化變化，藉助加工技術的開發，希望能有效的將其劣化的速度減緩或停止。此技術可能需藉助保鮮劑之效益，調控儲運過程中對表皮組織產生的乾燥變性、氧化反應及各項生化劣變以維持最大的商品價值及最少的經濟損耗。 (2)藉著酵素加工技術的利用有效的將不易分解利用的魷魚蛋白質資源轉化為具機能特性的小分子胜肽及胺基酸，分析此類產品的自由基清除效力、降血壓能力、抑制糖解酵素活性的表現及酪氨酸酶抑制活性。探討其各種機能特性表現與加工條件間之關係，應用加工條件之調控強化其機能性之表現，拓廣產品可能應用方向。 3.預期效益：（1）改善遠洋魷魚儲運品質，提升商品價格，穩定魷魚原料供應品質，確保原料安全衛生。（2）改善魷魚蛋白質資源的有效利用，提昇產品機能特性，拓展魷魚加工層次的優質及精緻化，確保漁民收益。
1. Program goals: (1) Frozen squid undergo deterioration during storage primary due to discoloration of skin. Improper way of handling and storage environment cause the reject of product and thus economic loss. The aim of this project is to develop effective treatment to keep squid quality without dehydration, oxidation and discoloration during frozen storage. (2)Squid are excellent marine food resources can provide a rich source of protein and high unsaturated fatty acids. However, the organizational structure of squid muscle protein properties, to withstand hot and dry will form a more flexible and hardness succulent organization, is not easy for human digestion and absorption. In order to effectively enhance the use of resources, can be developed to provide easily digested protein hydrolysis decomposition products processing. This is not only enhance its value-added, will also be upgraded to a health care function of the characteristics of food. Industrial level could be high-quality and refined to create more market competitiveness and economic strength. 2. Key projects: (1)This project focuses on gaining a better understanding on how the deterioration reaction occurs. Properly handling and food additives would be efficient for prevention squid from an inappropriate commercial value loss by dehydration, lipid oxidation and fading skin color. (2)Bioactive peptides can be released by enzymatic proteolysis of squid protein and the free radical scavenging activity, angiotensin-I-converting enzyme inhibitory activity α-amylase inhibitory activity, tyrosinase inhibitory activity were assayed. Controlled enzymatic hydrolysis of squid proteins may produce a series of small polypeptides which can modify and even improve functional properties and the application of squid proteins. 3. Anticipated benefits: (1)Squid storage quality improve and enhance the value of catch merchandise. Squid supply can keep quality and safety. (2)The effective development and utilization of resources and upgrade the squid protein functional properties of products. Industrial level could be high-quality and refined to create more market competitiveness and economic strength and to ensure that fishermen's income
The Innovative Research about the Development of Intelligent Solar Illumination System(I)
No full text研究領域:能源工程, 光電工程
計畫編號:NSC98-2622-E273-004-CC