成功大學博碩士論文系統

簡易檢索 / 詳目顯示

回結果列表

研究生：	廖偉茵 Liao, Wei-yin
論文名稱：	生理陽離子、三酸甘油酯與lipoprotein-associated phospholipase A2生化感測器的研究 Studies of physiological cations, triglycerides and lipoprotein-associated phospholipase A2 biosensors
指導教授：	劉炯權 Liu, Chung-Chiun 周澤川 Chou, Tse-Chuan
學位類別：	博士 Doctor
系所名稱：	工學院 - 化學工程學系 Department of Chemical Engineering
論文出版年：	2008
畢業學年度：	96
語文別：	英文
論文頁數：	158
中文關鍵詞：	離子感測器、生化感測器、三酸甘油酯、脂蛋白相關磷脂酶A2
外文關鍵詞：	Lipoprotein-associated phospholipase A2, biosensors, Triglycerides, Ion sensors
相關次數：	點閱：74 下載：2
分享至:	分享至facebook 分享至twitter

查詢本校圖書館目錄查詢臺灣博碩士論文知識加值系統勘誤回報

本論文之內容著重於兩種生化感測器的開發，分別為電位式離子感測器與電流式三酸甘油酯與脂蛋白相關磷脂酶A2感測器。這兩種感測器皆以微製造技術製備，以便達成隨時監控人體的狀況的目地，就像市售的血糖儀，人們可以隨時隨地量測自己的血糖一樣。這種儀器的發明使得人類能花費更少的時間就能得知身體狀況，因此開發適用於此種儀器的感測器亦發重要。相同的，能直接偵測心臟疾病的目標分子：三酸甘油酯與脂蛋白相關磷脂酶A2的生化感測器的開發更能替人類有效的預防疾病的發生。因此，本研究最主要的動機即為發展偵測生理陽離子，三酸甘油酯與脂蛋白相關磷脂酶A2生化感測器的晶片。
電位式離子感測器在本研究中有兩種不同的形式的晶片，分別為陣列式與微流體形式的晶片。
離子陣列式晶片，它是以一種可以量測生物樣品中酸鹼值、鉀、鈉、銨和鈣離子濃度的晶片。再開發這種晶片最重要的是發展穩定性佳的參考電極。因此，本研究闡述以網印電極方式來製造平版式的參考電極晶片，來取代傳統參考電極。此晶片式參考電極使用含有氯化鈉的洋菜膠為內電解質以及二氯稀橡膠作為液體接界和絕緣膠。結果顯示，此晶片式參考電極並不受多種生理上重要的離子的濃度影響。將此晶片式參考電極與酸鹼感測器與不同的離子電極結合便可形成一量測酸鹼值的晶片與陣列式離子感測晶片。結果顯示，酸鹼值檢測晶片與陣列式離子感測晶片的靈敏度與酸鹼感測器和離子電極相對於傳統參考電極所得的靈敏度相差不大。由此得知，晶片式參考電極的性能與傳統參考電極相當。
微流體式的晶片亦包含酸鹼值、鉀和鈣離子感測電極。此三種感測器分別為以氧化銥、鉀和鈣離子選擇膜修飾白金電極而成。此晶片包含運輸檢體的動力即微幫浦使檢體能在微流道中通興並流經感測器。結果顯示，這種電位式感測器在此微流頭體晶片中亦能有能斯特反應的能力，靈敏度趨近於理想值。
心臟病在整個人類族群中成為致命的疾病。近年來不僅三酸甘油酯而且脂蛋白相關磷脂酶A2同樣為心血管疾病重要的指標。因此，本研究開發可丟棄式以銥修飾的碳電極來感測血清中的三酸甘油酯與脂蛋白相關磷脂酶A2。感測的機制建立於量測酵素反應的產物：過氧化氫與尼克酰胺腺嘌呤二核苷酸。結果顯示，丁酸甲酯可作為量測脂蛋白相關磷脂酶A2的基直，在0到150個單位的脂蛋白相關磷脂酶A2其靈敏度為1.45 nAU-1。甘油三丁酸酯的量測中得到在0到10豪莫爾濃度其其靈敏度為7.5 nA(mM)-1。同時亦用尿酸及維生素C來評估此感測器，結果顯示影響甚微。最後本研究亦找出最佳的偵測條件於使可丟棄式生化感測器。

This dissertation is focused on two important types of biosensors, (1) potentiometric ion sensors and (2) amperometric biosensors for determining triglycerides (TG) and lipoprotein-associated phospholipase A2 (Lp-PLA2). Both of these kinds of sensors were fabricated by microfabrication technology to achieve the final goal of being useable tools for real-time monitoring of body chemistry. For example, the glucose meter was created for people to detect glucose levels in blood at anytime and anywhere. This kind of instrument ensures that people can spend less time to be concerned about their health than they used to be. There is no doubt that the sensors used in the glucose meter can provide a convenient way to assist people in measuring blood glucose. Hence, the development of the biosensors, which can detect the biomarkers of disease will help people predict the possibility of disease happing. For this reason, developing a chip having multiple sensors for the evaluation of significant physiological cations and biomarkers of diseases (Lp-PLA2 and TG) is the motivation of this research.
The potentiometric ion sensors in this study were created in two different forms i.e. as sensor array chips and microfluidic devices.
An ion sensor array chip is a chip which can measure the pH, K+, Na+, NH4+ and Ca2+ ion concentrations of biological samples. In creating this array, it is necessary to develop a chip type reference electrode to improve the practicability and preciseness. Here, the fabrication of a planar-form, chip type reference electrode using a screen-printing method was developed to replace a commercial reference electrode. The reference electrode chip uses agar gel as the inner electrolyte and chloroprene rubber for the liquid junction and insulator. It was shown that the reference electrode chip is insensitive to most of the physiologically important ionic species. Integration of the reference electrode chip with four different ion selective electrodes (potassium, sodium, ammonium and calcium) and a pH indicator, on a substrate, to form an ion sensor array chip was performed. There was no significant difference between the sensitivity obtained from ion selective electrodes employing our reference electrode chip and from ion selective electrodes formed with a commercial reference electrode. These results indicate that our fabricated reference electrode was at least equal to commercial reference electrodes, in terms of sensitivity, while being compatible with potentiometric ion sensors fabricated on the same substrate as in an ion sensor array.
A microfluidic device with an all-solid-state potentiometric biosensor array was developed by using microfabrication technology. The sensor array included a pH indicator, potassium and calcium ion-selective microelectrodes. The detection system was integrated with a micro-pneumatic pump which can continuously drive fluids into the microchannel through sensors. The sensor array microfluidic device showed near Nerstian responses.
Heart disease has become a major heath concern for entire population. Recently, not only TG but Lp-PLA2 was also an important marker of cardiovascular disease. In this study, detection and quantification of Lp-PLA2 and TG using an iridium modified carbon based biosensor were successfully carried out. The detection procedure was based on measuring the enzymatically produced hydrogen peroxide and NADH from the reactions of methyl ester hydrolysis by Lp-PLA2 and glycerol oxidation by glycerol dehydrogenase. From the results, methyl butyrate can be as the substrate for Lp-PLA2 assay. The detection of Lp-PLA2, in the concentration range 0 to 150 Uml-1, was established as following a linear relationship with a sensitivity of 1.45 nAU-1 in bovine serum. A linear response to glyceryl tributyrate in the concentration range of 0 to 10 mM and a sensitivity of 7.5 nA(mM)-1 in bovine serum. The potential interference of species such as uric acid (UA) and ascorbic acid (AA) was assessed. The incorporation of a selected surfactant and an increase in the incubation temperature appeared to enhance the performance of this biosensor. The conditions for the determination of Lp-PLA2 and TG levels in bovine serum using this biosensor were optimized.

Table of the Contents
English abstract	I
Chinese abstract	IV
Table of the contents	VI
List of Tables  	X
List of Figures	        XI

Chapter 1 Introduction	1
1.1  Reference electrodes in electrochemistry	1
   1.1.1  Reference electrodes of second kind	5
   1.1.2  Criteria for reference electrodes	6
   1.1.3  Microfabricated reference electrodes	11
   1.1.4  The created reference electrode on the chip	12
1.2  pH indicator	13
   1.2.1  General principals of metal-metal oxide electrode	14
   1.2.2  Metal oxide pH sensors	16
1.3  Ion-selective electrode and its response mechanism	17
1.3.1  Chip-type pH sensor and ion sensor array	22
1.4  pH and Ion-selective electrodes in microfluidic system	22
1.4.1  Fluid driving: micropump for ion-sensor array microfluidic chip	23
   1.4.2  Pneumatic micropump	25
1.5  The biology of Atherosclerosis	27
1.5.1  The structure of normal human artery	30
1.5.2  Endothelial dysfunction	32
1.5.3  Stages of atherosclerosis	33
1.5.4  Biomarker of heart disease: Lp-PLA2	36
1.5.5  Structural and catalytic properties of Lp-PLA2	38
1.5.6  Biomarker of heart disease: triglyceride	39
1.5.7  Structure of triglyceride	40
1.5.8  Lp-PLA2 and triglyceride assays by biosensors	42
Chapter 2 Principals 	44
2.1  The mechanism of reference electrode on the chip	44
2.2  The mechanism of iridium oxide pH indicator	44
2.3  Theory of methanol evaluation by chemical reagents	46
2.4  Mechanistic basis of Lp-PLA2 sensing	48
2.5  Mechanistic basis of triglyceride sensing	49
Chapter 3 Experimental	52
3.1  Chemicals, materials, and instruments	52
3.2  Experimental Techniques	56
   3.2.1  Fabrication procedures of the screen-printed solid electrolyte modified Ag/AgCl reference electrode	56
	
       3.2.1.1  Evaluation of the screen-printed solid electrolyte modified Ag/AgCl reference electrode	58
	
3.2.2  Fabrication procedures of the pH indicator electrode	58
3.2.3  pH biosensor manufacture	60
3.2.3.1  Evaluation of pH indicator electrode and pH biosensor	63
	
3.2.4  Ion sensor array chip structure and fabrication	64
3.2.4.1  Evaluation of ion sensor array chip	67
   3.2.5  Ion sensor array microfluidic device structure and fabrication	67
	
       3.2.5.1  Device description	67
       3.2.5.2  Fabrication process of microfluidic device	69
       3.2.5.3  Fabrication process of microelectrodes	70
       3.2.5.4  Evaluation of ion sensor array microfluidic device	71
	
3.2.6  Disposable mini biosensor manufacture	72
       3.2.6.1  Preparation of the iridium nanoparticles dispersed carbon ink	72
	
       3.2.6.2  Screen-printing procedure of disposable min biosensor	72
        3.2.6.3  The procedures of using disposable min biosensor in Lp-PLA2 level assay	73
	
        3.2.6.4  The procedures of using disposable min biosensor in TG level assay	76
	
        3.2.6.5  The procedures of ascorbic acid and uric acid interfering investigation in detection of Lp-PLA2 and TG levels by disposable min biosensor	76
	
3.2.7  Methanol assay	78
        3.2.7.1  Preparation of methanol assay reagents	78
        3.2.7.2  Procedures of methanol assay by using methanol reagents	79
	
3.2.8  Safety note	79
Chapter 4 Results and discussion	80
4.1  The characteristics of reference electrode on the chip	80
4.1.1  The stability and durability of the reference electrode chip	84
	
4.1.2  Fabrication reproducibility of the reference electrode chip	87
	
4.1.3  Analytical applications of the reference electrode chip---pH sensor	89
	
4.1.4  In vitro measurement results of developed chip-type pH biosensor	89
	
4.1.5  Analytical applications of the reference electrode chip in ion sensor array measurements	91
	
4.2.1  Performance of the micropump	94
4.2.2  Response of potentiometric sensors array in the flow injection mode	99
	
4.2.3  Simultaneous responses of the sensors array	103
4.3.1  The performance of biosensor of detecting enzymatically produced hydrogen peroxide and NADH	109
	
4.3.2  Characterization of the Lp-PLA2 sensing mechanism	115
4.3.3  Evaluation of proposed mechanism to determine of Lp-PLA2 levels by a biosensor	117
	
4.3.4  Substrate selection for the Lp-PLA2 assay	120
4.3.5  Amperometrical Lp-PLA2 level assay and the influence of ascorbic acid and uric acid in this measurement	121
	
4.3.6  The surfactant effect in Lp-PLA2 level assay	126
4.3.7  Characterization of TG biosensors	128
4.3.8  Effects of ascorbic acid and uric acid on TG sensing	133
4.3.9  The Influences of surfactant and incubation temperature on TG sensing	135
	
4.3.10  Estimation of triglycerides	139
Chapter 5 Conclusions	143
Chapter 6 Suggestions	147
References	148
 
List of Tables
Table 1-1	Standard potentials for different reference electrodes of the second kind vs. SHE at 25 oC. The electrode processes in each case incorporate the dissociation reaction of the sparingly soluble salt.	9
		
Table 1-2 	Performance of iridium oxide-based pH electrodes made by different methods.	18
Table 3-1	Information of the chemicals and materials used in this study.	52
Table 3-2	Information of the instruments used in this study	55
Table 4-1 	In vitro pH measurement results obtained using developed pH biosensor and commercial pH meter.	93
Table 4-2	Sensitivities of pH indicator and calcium and potassium ion-selective electrodes in a beaker system and in the microfluidic device at 35μlmin-1.	101
Table 4-3 	Selectivity coefficients determined for calcium and potassium ion-selective electrodes in beaker system and in the microfluidic device at 35μl/min.	
102
Table 4-4 	Response times of pH indicator and calcium and potassium ion-selective electrodes in the microfluidic device (A) and in beaker system (B).	107



 
List of Figures
Figure 1-1	Simple forms of hydrogen electrode: (a) Hildebrand electrode; (b) and (c) the different modifications of Hildebrand electrode.	3
Figure 1-2	Construction of the measurement system for measure the potential of Cu by using SHE as reference electrode.	4
Figure 1-3	(A) Standard laboratory reference electrode. a; electrode head. b; reference system. c; reference electrolyte. d; electrode stem. e; filler. f; separator. (B) Commercial reference electrode from Bioanalytical systems Inc. 1; electrolyte. 2; Ag/AgCl. 3; porous ceramic frit.	7
Figure 1-4	Scientific forms of the calomel electrode: (a) standard from; (b) form simplified from standard type; (c) from suitable for technical measurements.	8
Figure 1-5	Experimental assembly for measuring ISEs in a zero current galvanic cell.	20
Figure 1-6	Basic cell notation of ISE cell assembly. Numbers and vertical bars denote boundary potentials. The sum of all individual boundary potentials make up the observed cell potential.	20
Figure 1-7	Classification of micropumps with different actuation methods.	24
Figure 1-8 	(Ⅰ): A) Plexiglas microchip module. Dimensions: 40 mm x 40 mm x 19.175 mm. B) Schematic diagram of the plexiglas microchip module. C) Cross-section of the mounting of a glass-silicon microchip on the microchip module showing the major components. (Ⅱ): Operating principles of a semi-disposable CNC (computer numerical control) machined plexiglass microvalve system. (a) valve open, (b) valve closed and (c) bottom view.	26
Figure 1-9	Cross-section and top views of three-layer (A) and four-layer (B) monolithic pneumatic PDMS membrane microvalves.	28
Figure 1-10	Schematic diagram of two-step micro RT-PCR chip integrated with micropumps and microvalves for controlling biosample transportation.	29
Figure 1-11	Structure of a normal large artery. This illustration displays the three distinct layers of the vessel wall: intima, media, and adventitia as well as the endothelium and the external internal elastic lamina.	31
Figure 1-12	Migration of monocytes into the arterial intima during the early stages of atherogenesis.	34
Figure 1-13	Participation of inflammation in all stages of atherosclerosis. A: initiation of atherosclerosis. B: progression of atherosclerosis. C: thrombotic complication of atherosclerosis	35
Figure 1-14	Lp-PLA2 in atherosclerosis-specific inflammatory pathways.	37
Figure 1-15	Structure of a triacylglycerol (triglyceride). The mixed triacylglycerol shown here has three different fatty acids attached to the glycerol backbone. When glycerol has two different fatty acids at C-1 and C-3, the C-2 is a chiral center.	41
Figure 2-1	Schematic drawing of the mechanisms of reference electrode chip and the commercial reference electrode.	45
Figure 2-2	(a) Reaction of the Schiff's reagent with an aldehdye (b) methanol oxidation by potassium permanganate (C) de-colorization reactions of potassium permanganate.	47
		
Figure 2-3	Schematic diagram showing the hydrolysis of oxidized phospholipids by Lp-PLA2 and the associated mechanism for the determination of Lp-PLA2 levels by the disposable mini biosensor. PC (phosphatidylcholine), PUFA (polyunsaturated fatty acid), ox-PC (oxidatively-modified phosphatidylcholine), ox-FA (oxidized fatty acid) and Lyso-PC (lysophosphatidylcholine).	50
Figure 2-4	Schematic diagrams of the mechanism of TG determination by the iridium modified carbon biosensors.	51
Figure 3-1	Schematic diagrams of the screen-printed solid electrolyte modified Ag/AgCl reference electrode and the right hand side is its cross-sectional view. Ⅰ: alumina substrate. Ⅱ: Ag electrode. Ⅲ: Ag/AgCl. Ⅳ: agar gel. Ⅴ: chloroprene rubber.	57
Figure 3-2	Schematic diagram of potentiometric measurement using two-electrode-system setup for evaluation of solid electrolyte modified Ag/AgCl reference electrode. The electrochemical working-shop has three ports including: working electrode (green line), counter electrode (red line) and reference electrode (yellow line). (For two-electrode system, red line and yellow line have to connect together with a commercial reference electrode.)	59
Figure 3-3	Schematic diagrams of the pH indicator and the right hand side is its cross-sectional view. Ⅰ: alumina substrate. Ⅱ: Pt electrode. Ⅲ: iridium oxide. Ⅳ: insulator.	61
Figure 3-4	Schematic diagrams of formation processes of the pH sensor. The right hand side is the cross-sectional view of finished pH sensor. 	62
Figure 3-5	Schematic diagrams of formation processes of ion sensor array chip. (A) A complete structure of ion sensor array chip. (B) Procedure of formation electronic circuit of ion sensor array chip. (C) Formation of reference electrode and (D) ion detectors of the ion sensor array chip.	65
Figure 3-6	A three-dimensional schematic representation of the ion sensor array microfluidic device.	68
Figure 3-7	The photograph of the disposable mini biosensor.	74
Figure 3-8	The flowchart of PLA2 level measurement by a biosensor.	75
Figure 3-9	The flowchart of triglyceride measurement by a biosensor.	77
Figure 4-1	(a) The dynamic emf response curves of the reference electrode chip versus a commercial Ag/AgCl reference electrode in various chloride ion concentration solutions. (b) The dynamic emf response curves of Ag/AgCl electrode versus a commercial Ag/AgCl reference electrode in various potassium chloride concentration solutions. (c) Typical calibration curve of Ag/AgCl electrode versus a commercial Ag/AgCl reference electrode in various potassium chloride concentration solutions.	82
Figure 4-2	SEM images of chloroprene rubber. (a) They were captured at two different places of top side (the surface faces to the sample solution). (b) They were taken at two different places of bottom (the surface faces to the agar gel) side.	83
Figure 4-3	Stability tests of the reference electrode chip in unstirred 10-6 M (▓), 10-4 M (○), 10-2 M (△), 1 M (＋), and 3 M (◇) KCl solutions.	85
Figure 4-4	The dynamic emf response curves of the reference electrode chip versus a commercial reference electrode with continuously varying concentrations of nitrate, acetate, carbonate, phosphate, and sulfate and in different pH buffer solutions. Operation temperature: 25 oC ± 3 oC. Solution volume: 30 ml.	86
Figure 4-5	The dynamic emf response curves of 10 reference electrode chips versus a commercial reference electrode in continuously changed KCl concentrations. Operation temperature: 25 oC ± 3 oC. Solution volume: 30 ml.	88
Figure 4-6	Stability test of reference electrode chip versus a commercial reference electrode in different pH solutions (a). Response curves of an iridium oxide modified Pt pH indicator electrode versus a commercial reference electrode (b) and reference electrode chip (c). These curves were obtained by titrating 1 M KOH solutions against 0.1 M KNO3-0.01 M H3PO4-H3BO3-CH3COOH and the pH of the test solution was varied from 2.00 to 10.11. Operation temperature: 25 oC ± 3 oC. Solution volume: 30 ml.	90
Figure 4-7	Potential response curves of pH indicator with commercial reference electrode (solid line) and developed pH biosensor (dashed line) in: (Ⅰ): standard buffer pH=4.01, (Ⅱ): standard buffer pH=7.01, (Ⅲ): standard buffer pH=10.01, (Ⅳ): Coca Cola, (Ⅴ): tap water, (Ⅵ): orange juice, (Ⅶ): milk (low fat), (Ⅷ): human serum, and (Ⅸ): human whole blood. Operation temperature: 25 oC ± 3 oC.	92
Figure 4-8	The reference electrode chip integrated with ammonium (a), potassium (b), sodium (c) and calcium (d) ion-selective electrodes. The solid lines in each figure (from a to d) were emf responses of ion-selective electrodes versus a commercial reference electrode with stepwise concentration changes of NH4Cl, KCl, NaCl and CaCl2 solutions from 10-8 M to 0.3 M. The dash lines shows the potential responses of ion-selective electrodes versus the reference electrode chip, ion sensor array, with stepwise concentration changes in the NH4Cl, KCl, NaCl and CaCl2 solutions. Operation temperature: 25 oC ± 3 oC. Solution volume: 30 ml.	97
Figure 4-9	Schematic representation of the peristaltic activation of three sets of thin films to pump the fluids to flow in specific direction. (B) Relationship between pumping rate and driving frequency for the peristaltic pneumatic micropump. The maximum pumping rate is 52.4μlmin-1 at a frequency of 36 Hz with the pressure 10 psi.	98
Figure 4-10	Calibration plots determined individually for pH indicator, calcium and potassium ion-selective electrodes in the microfluidic device operated in flow injection potentiometry mode at 35μlmin-1. The calibration solution made as 0.1 M KNO3 - 0.01 M H3PO4-H3BO3-CH3COOH and adjusted to the desired pH with 1 M KOH or 1 M HNO3 as required was injected into the device. KNO3 and Ca(NO3)2 stock solutions were diluted as necessary to give a serious of calibration solutions ranging from 10-1 M to 10-8 M.	100
Figure 4-11	Typical simultaneous response curves of pH indicator, calcium and potassium ion-selective electrodes in the microfluidic device at 35μlmin-1.	105
Figure 4-12	Sensitivities of pH indicator, calcium and potassium ion-selective electrodes at different flow rates in the microfluidic device.	106
Figure 4-13	The cyclic voltammorgrams of the biosensor in the presence of different amount of hydrogen peroxide in PBS (pH=7.4) buffer solution. Scan rate: 10 mV/sec. Operation temperature: 25 oC. Solution volume: 10 ml. Only one biosensor was used. CVs were obtained without stirring	110
Figure 4-14	The cyclic voltammorgrams of the biosensor in the presence of different amount of methanol in PBS (pH=7.4) buffer solution. Alcohol oxidase: 10U. Incubation time: 1 min. Scan rate: 10 mV/sec. Operation temperature: 25 oC. Solution volume: 1 ml. Only one biosensor was used. CVs were obtained without stirring.	111
Figure 4-15	(A) Chronoamperograms of the biosensor to the different activities of methanol. (B) calibration plot for the mini-biosensor in different methanol concentrations with PBS (pH=7.4) buffer solution, which contained 10 U alcohol oxidase. The applied potential was 0.2 V vs. printed Ag/AgCl. Alcohol oxidase: 10U. Incubation time: 1 min. Operation temperature: 37 ±3 oC. Solution volume: 1 ml. Stead steady state current response curves were obtained without stirring.	113
Figure 4-16	The cyclic voltammorgrams of the biosensor in the presence with and without glycerol in PBS (pH=7.4) buffer solution. Glycerol dehydrogenase: 8U. Substrate: 0.1 M NAD+. Incubation time: 1 min. Scan rate: 10 mV/sec. Operation temperature: 25 oC. Solution volume: 1 ml. Only one biosensor was used. CVs were obtained without stirring.	114
Figure 4-17	(A) Chronoamperograms of the biosensor to the different activities of glycerol. (B) Glycerol calibration: current measured at fixed time (20 s) versus different glycerol concentrations. The applied potential was 0.15 V vs. printed Ag/AgCl. Glycerol dehydrogenase: 8U. Substrate: 0.1 M NAD+. Incubation time: 1 min. Operation temperature: 25 oC. Solution volume: 1 ml. Stead steady state current response curves were obtained without stirring.
                Only one biosensor was used.	116
Figure 4-18	UV spectrograms of: (a) phosphate buffer pH 7.0, (b) 0.01 M methanol aqueous solution, (c) 0.01 M butyric acid solution, (d) pure PLA2 solution and (e) the hydrolysis solution consisting of 50 μl methyl butyrate in 1ml PLA2 solution (incubated for 2 hours), each of the above was mixed with the methanol assay reagents using the procedures described in section 3.5.	118
Figure 4-19	Cyclic voltammorgrams of the biosensor in the presence of different amounts of PLA2 in 0.4 M methyl butyrate in bovine serum solutions which additionally contained 5U of alcohol oxidase. This experiment was accomplished using one biosensor and 90 seconds was used for incubating the reaction after each addition of PLA2. The scan rate was 10 mV/sec. The operation temperature was 37 ±3 oC.	119
Figure 4-20	Effects of the substrate’s carbon chain length in Lp-PLA2 level assay. Changes in current were obtained in 1 ml of 0.4 M methyl acetate, methyl butyrate, methyl hexanoate and methyl octanoate in bovine serum. Each serum sample contained alcohol oxidase (5U) and PLA2 (25U). The reaction was incubated for 90 sec. The applied potential was 0.2V versus printed Ag/AgCl and the operating temperature was 37 ±3 oC. The error bars represent the data obtained from four individual min biosensors used for each substrate analysis.	122
Figure 4-21	(a) The steady state current response curves taken from a single biosensor in response to different PLA2 levels. The test sample was 1ml of 0.4 M methyl butyrate in bovine serum solution which contained 5U of alcohol oxidase. The applied potential was 0.2V versus printed Ag/AgCl and the operating temperature was 37 ±3 oC. (b) The corresponding calibration plot obtained after 10 seconds on the steady state current response curves from 0 to 150 U (the error bars represent data for each PLA2 level obtained from three new individual biosensors together with the biosensor used for steady state current response curves. Therefore, for standard deviation n=4.).	123
Figure 4-22	(a) Effect of uric acid (UA) and ascorbic acid (AA) on PLA2 activity calibration curves. Currents versus PLA2 activity were obtained in 0.4 M methyl butyrate in bovine serum (■), containing 416 μmoleL-1 UA (▲) and 86 μmoleL-1 AA (●). (b) The calibration curves after elimination of the background currents. The test sample for each data point additionally contained 5U of alcohol oxidase. The incubation time was 90 seconds. The applied potential was 0.2V versus printed Ag/AgCl and the operating temperature was 37 ±3 oC.	125
Figure 4-23	Effect of the surfactant triton x 100 quantity contained in 0.4 M methyl butyrate bovine serum solutions of the PLA2 activity determination. The solution for each data point contains 5U of alcohol oxidase and 25 U of PLA2. The applied potential is 0.2V versus printed Ag/AgCl.	127
Figure 4-24	(a) Chronoamperograms of the biosensor to the different activities of PLA2 in 0.4 M methyl butyrate bovine serum solutions which contained 0.25% triton X-100 (v/v) and 5U of alcohol oxidase. (b) PLA2 calibration: current measured at fixed time (10 s) vs enzyme activity. The applied potential is 0.2V versus printed Ag/AgCl.	129
Figure 4-25	(a) Cyclic voltammorgrams of the biosensor with different amounts of glycerol in bovine serum containing NAD+ (0.01 M) and GDH (8U). (b) cyclic voltammorgrams of the biosensor with different amounts of glyceryl tributyrate in bovine serum with the presence of lipase (10 mg), NAD+ (0.01 M) and GDH (8U). The scan rates of (a) and (b) were controlled at 10 mV/sec.	130
Figure 4-26	(a) Chronoamperograms of the biosensor to different concentrations of glyceryl tributyrate. The applied potential was 0.15V versus printed Ag/AgCl. (b) The calibration curve was plotted using the current measured at fixed time interments (10th s) versus glyceryl tributyrate concentrations. The serum volume was 1ml containing lipase (10 mg), NAD+ (0.01 M) and GDH (8U). 	132
Figure 4-27	(a) Effect of uric acid and ascorbic acid on glyceryl tributyrate calibration curves. Currents vs. different glyceryl tributyrate concentrations were obtained in bovine serum solutions (■), containing 416 μmoleL-1 UA (▲) and 86 μmoleL-1 AA (●). (b) The calibration curves after background current elimination. The serum (1ml), addtionally contained lipase (10 mg), NAD+ (0.01 M) and glycerol dehydrogenase (8U). The applied potential was 0.15V versus printed Ag/AgCl.	134
Figure 4-28	Effect of the surfactant triton x 100 quantity contained of the biosensor in 2 mM glyceryl tributyrate bovine serum. The samples for each data point contained lipase (10 mg), NAD+ (0.01 M) and GDH (8U). The applied potential was 0.15V versus printed Ag/AgCl.	136
Figure 4-29	Effect of the incubation temperture of the biosensor in evaluation of 2 mM glyceryl tributyrate bovine serum. The samples for each data point contained lipase (10 mg), NAD+ (0.01 M) and GDH (8U). The applied potential was 0.15V versus printed Ag/AgCl.	138
Figure 4-30	Cyclic voltammorgrams of the biosensor with different amounts of sunflower seed oil in the presence of lipase (10 mg), NAD+ (0.01 M) and GDH (8U) bovine serum. The scan rate was controlled at 10 mV/sec.	140
Figure 4-31	(a) Chronoamperograms of the biosensor to the different concentrations of sunflower oil. The applied potential was 0.15V versus printed Ag/AgCl. (b) The calibration curve was plotted using the current measured at fixed time (10th s) interments vs. different sunflower seed oil concentrations. The samples for each curves was 1ml of bovine serum, which additionally contained lipase (10 mg), NAD+ (0.01 M) and GDH (8U).	141
                                    

References
[1] A.J. Bard, L.R. Faulkner, Electrochemical methods: fundamentals and applications, John Wiley, New York, 2001.
[2] D.J.G. Ives, G.J. Janz, Reference electrodes, theory and practice, Academic Press, New York,, 1961.
[3] D. Ammann, Ion-selective microelectrodes: Principles, Design and Application, Springer-Verlag, Berlin, New York 1986.
[4] L.A. Kaplan, A.J. Pesce, S.C. Kazmierczak, Clinical chemistry: theory, analysis, correlation, Mosby, St. Louis, 2003.
[5] T.H. Gieling, G. van Straten, H.J.J. Janssen, H. Wouters, ISE and chemfet sensors in greenhouse cultivation, Sens. Actuators B-Chem., 105 (2005) 74-80.
[6] S. Martinoia, G. Massobrio, L. Lorenzellib, Modeling ISFET microsensor and ISFET-based microsystems: a review, Sens. Actuators B-Chem., 105 (2005) 14-27.
[7] C.H. Hamann, W. Vielstich, A. Hamnett, Electrochemistry, Wiley-VCH, Weinheim, 2007.
[8] J.S. Newman, K.E. Thomas-Alyea, Electrochemical systems, Wiley-Interscience, Hoboken, N.J., 2004.
[9] D. Diamond, E. McEnroe, M. McCarrick, A. Lewenstam, Evaluation of a new solid-state reference electrode junction material for ion-selective electrodes, Electroanalysis, 6 (1994) 962-971.
[10] L. Tymecki, Z. Zwierkowska, R. Koncki, Screen-printed reference electrodes for potentiometric measurements, Anal. Chim. Acta, 526 (2004) 3-11.
[11] A.W.J. Cranny, J.K. Atkinson, Thick film silver silver chloride reference electrodes, Meas. Sci. Technol., 9 (1998) 1557-1565.
[12] P.J. Kinlen, J.E. Heider, D.E. Hubbard, A Solid-State Ph Sensor-Based on a Nafion-Coated Iridium Oxide Indicator Electrode and a Polymer-Based Silver-Chloride Reference Electrode, Sens. Actuators B-Chem., 22 (1994) 13-25.
[13] M.A. Nolan, S.H. Tan, S.P. Kounaves, Fabrication and characterization of a solid state reference electrode for electroanalysis of natural waters with ultramicroelectrodes, Anal. Chem., 69 (1997) 1244-1247.
[14] H.R. Kim, Y.D. Kim, K.I. Kim, J.H. Shim, H. Nam, B.K. Kang, Enhancement of physical and chemical properties of thin film Ag/AgCl reference electrode using a Ni buffer layer, Sens. Actuators B-Chem., 97 (2004) 348-354.
[15] K. Eine, S. Kjelstrup, K. Nagy, K. Syverud, Towards a solid state reference electrode, Sens. Actuators B-Chem., 44 (1997) 381-388.
[16] K. Nagy, K. Eine, K. Syverud, O. Aune, Promising new solid-state reference electrode, J. Electrochem. Soc., 144 (1997) L1-L2.
[17] H.J. Lee, U.S. Hong, D.K. Lee, J.H. Shin, H. Nam, G.S. Cha, Solvent-processible polymer membrane-based liquid junction-free reference electrode, Anal. Chem., 70 (1998) 3377-3383.
[18] H.J. Yoon, J.H. Shin, S.D. Lee, H. Nam, G.S. Cha, T.D. Strong, R.B. Brown, Solid-state ion sensors with a liquid junction-free polymer membrane-based reference electrode for blood analysis, Sens. Actuators B-Chem., 64 (2000) 8-14.
[19] E. Bakker, Hydrophobic membranes as liquid junction-free reference electrodes, Electroanalysis, 11 (1999) 788-792.
[20] Y.M. Mi, S. Mathison, E. Bakker, Polyion sensors as liquid junction-free reference electrodes, Electrochem. Solid State, 2 (1999) 198-200.
[21] H. Suzuki, T. Hirakawa, S. Sasaki, I. Karube, Micromachined liquid-junction Ag/AgCl reference electrode, Sens. Actuators B-Chem., 46 (1998) 146-154.
[22] H. Suzuki, A. Hiratsuka, S. Sasaki, I. Karube, Problems associated with the thin-film Ag/AgCl reference electrode and a novel structure with improved durability, Sens. Actuators B-Chem., 46 (1998) 104-113.
[23] H. Suzuki, T. Hirakawa, S. Sasaki, I. Karube, An integrated three-electrode system with a micromachined liquid-junction Ag/AgCl reference electrode, Anal. Chim. Acta, 387 (1999) 103-112.
[24] H. Suzuki, T. Taura, Thin-film Ag/AgCl structure and operational modes to realize long-term storage, J. Electrochem. Soc., 148 (2001) E468-E474.
[25] T. Matsuo, H. Nakajima, Characteristics of reference electrodes using a polymer gate ISFET, Sens. Actuators, 5 (1984) 293-305.
[26] H. Nakajima, M. Esashi, T. Matsuo, The cation concentration response of polymer gate ISFET, J. Electrochem. Soc., 129 (1982) 141-143.
[27] M. Fujihira, M. Fukui, T. Osa, Chemically modified pararylene gate field-effect transistor-preparation of pH insensitive parylene gate for chemical modofication, J. Electroanal. Chem., 106 (1980) 413-418.
[28] H. Perrot, N. Jaffrezicrenault, N.F. Derooij, H.H. Vandenvlekkert, Ionic detection using differential measurement between an ion-selective FET and a reference FET, Sens. Actuators, 20 (1989) 293-299.
[29] J.M. Chovelon, J.J. Fombon, P. Clechet, N. Jaffrezicrenault, C. Martelet, A. Nyamsi, Y. Cros, Sensitization of dielectric surfaces by chemical grafting-application to pH ISFETS and REFETS, Sens. Actuators B-Chem., 8 (1992) 221-225.
[30] A. Simonis, H. Luth, J. Wang, M.J. Schoning, New concepts of miniaturised reference electrodes in silicon technology for potentiometric sensor systems, Sens. Actuators B-Chem., 103 (2004) 429-435.
[31] N. Oyama, T. Ohsaka, S. Ikeda, K. Okuaki, A solid-state reference electrode based on bilayer coating with poly(p,p'-biphenaol) and polyimide films for the gate of field-effect transistor, Anal. Sci., 5 (1989) 729-734.
[32] W. Potter, C. Dumschat, K. Cammann, Miniaturized reference electrode based on a perchlorate-selective field-effect transistor, Anal. Chem., 67 (1995) 4586-4588.
[33] W.Y. Liao, T.C. Chou, Fabrication of a planar-form screen-printed solid electrolyte modified Ag/AgCl reference electrode for application in a potentiometric biosensor, Anal. Chem., 78 (2006) 4219-4223.
[34] S. Glab, A. Hulanicki, G. Edwall, F. Ingman, Metal-metal oxide and metal-oxide electrodes as pH sensors, Crit. Rev. Anal. Chem., 21 (1989) 29-47.
[35] G. Edwall, Improved Antimony-Antimony(Iii) Oxide Ph Electrodes, Med. Biol. Eng. Comput., 16 (1978) 661-669.
[36] R.E.F. Einerhand, W.H.M. Visscher, E. Barendrecht, pH measurement in strong KOH solutions with a bismuth electrode, Electrochim. Acta, 34 (1989) 345-353.
[37] C.C. Liu, B.C. Bocchicchio, P.A. Overmyer, M.R. Neuman, Palladium-palladium oxide miniature pH electrode, Science, 207 (1980) 188-189.
[38] W.T. Grubb, L.H. King, Palladium-palladium oxide pH electrodes, Anal. Chem., 52 (1980) 270-273.
[39] A. Fog, R.P. Buck, Electronic semiconducting oxide as pH sensors, Sens. Actuator, 5 (1984) 137-146.
[40] K. Pasztor, A. Sekiguchi, N. Shimo, N. Kitamura, H. Masuhara, Electrochemically-deposited RuO2 films as pH sensors, Sens. Actuators B-Chem., 14 (1993) 561-562.
[41] H.N. McMurray, P. Douglas, D. Abbot, NOVEL Thick-film pH sensors based on ruthenium dioxide glass composites, Sens. Actuators B-Chem., 28 (1995) 9-15.
[42] S. Yao, M. Wang, M. Madou, A pH electrode based on melt-oxidized iridium oxide, J. Electrochem. Soc., 148 (2001) H29-H36.
[43] S.A.M. Marzouk, S. Ufer, R.P. Buck, T.A. Johnson, L.A. Dunlap, W.E. Cascio, Electrodeposited iridium oxide pH electrode for measurement of extracellular myocardial acidosis during acute ischemia, Anal. Chem., 70 (1998) 5054-5061.
[44] G. Papeschi, S. Bordi, C. Beni, L. Ventura, Use of an iridium electrode for direct measurement of PI of proteins after ioselectric-focusing in polyacrylasmide-gel, Biochimica Et Biophysica Acta, 453 (1976) 192-199.
[45] P.J. Kinlen, J.E. Heider, D.E. Hubbard, A Solid-state pH sensors-based on a nafion-coated iridium oxide indicator electrode and a polymer-based silver-chloride reference electrode, Sens. Actuators B-Chem., 22 (1994) 13-25.
[46] I. Song, K. Fink, J.H. Payer, Metal oxide metal pH sensor: Effect of anions on pH measurements, Corrosion, 54 (1998) 13-19.
[47] J.E. Baur, T.W. Spaine, Electrochemical deposition of iridium(IV) oxide from alkaline solutions of iridium(III) oxide, J. Electroanal. Chem., 443 (1998) 208-216.
[48] A.J. Bard, M. Stratmann, Encyclopedia of electrochemistry, Wiley-VCH, Weinheim ; Cambridge, 2002.
[49] E. Bakker, P. Buhlmann, E. Pretsch, Carrier-based ion-selective electrodes and bulk optodes. 1. General characteristics, Chem. Rev., 97 (1997) 3083-3132.
[50] P. Buhlmann, E. Pretsch, E. Bakker, Carrier-based ion-selective electrodes and bulk optodes. 2. Ionophores for potentiometric and optical sensors, Chem. Rev., 98 (1998) 1593-1687.
[51] S.H. Wang, T.C. Chou, C.C. Liu, Development of a solid-state thick film calcium ion-selective electrode, Sens. Actuators B-Chem., 96 (2003) 709-716.
[52] S.H. Wang, T.C. Chou, Immobilized ionophore calcium ion sensor modified by montmorillonite, Electroanalysis, 12 (2000) 468-470.
[53] C.S. Zhang, D. Xing, Y.Y. Li, Micropumps, microvalves, and micromixers within PCR microfluidic chips: Advances and trends, Biotechnol. Adv., 25 (2007) 483-514.
[54] S.J. Lee, S.Y. Lee, Micro total analysis system (mu-TAS) in biotechnology, Appl. Microbiol. Biotechnol., 64 (2004) 289-299.
[55] A. Nisar, N. AftuIpurkar, B. Mahaisavariya, A. Tuantranont, MEMS-based micropumps in drug delivery and biomedical applications, Sens. Actuators B-Chem., 130 (2008) 917-942.
[56] H.T.G. Vanlintel, F.C.M. Vandepol, S. Bouwstra, A piezoelectric micropump based on micromachining of silicon, Sens. Actuators B-Chem., 15 (1988) 153-167.
[57] J.G. Smits, Piezoelectric micropump with 3 valves working peristaltically, Sens. Actuator A-Phys., 21 (1990) 203-206.
[58] D.J. Laser, J.G. Santiago, A review of micropumps, J. Micromech. Microeng., 14 (2004) R35-R64.
[59] P. Woias, Micropumps - past, progress and future prospects, Sens. Actuators B-Chem., 105 (2005) 28-38.
[60] R. Zengerle, J. Ulrich, S. Kluge, M. Richter, A. Richter, A bidirectional silicon micropump, Sens. Actuator A-Phys., 50 (1995) 81-86.
[61] T. Bourouina, A. Bosseboeuf, J.P. Grandchamp, Design and simulation of an electrostatic micropump for drug-delivery applications, J. Micromech. Microeng., 7 (1997) 186-188.
[62] M.M. Teymoori, E. Abbaspour-Sani, Design and simulation of a novel electrostatic peristaltic micromachined pump for drug delivery applications, Sens. Actuator A-Phys., 117 (2005) 222-229.
[63] A. Machauf, Y. Nemirovsky, U. Dinnar, A membrane micropump electrostatically actuated across the working fluid, J. Micromech. Microeng., 15 (2005) 2309-2316.
[64] S. Bohm, W. Olthuis, P. Bergveld, A plastic micropump constructed with conventional techniques and materials, Sens. Actuator A-Phys., 77 (1999) 223-228.
[65] C. Yamahata, C. Lotto, E. Al-Assaf, M.A.M. Gijs, A PMMA valveless micropump using electromagnetic actuation, Microfluid. Nanofluid., 1 (2005) 197-207.
[66] C. Yamahata, M. Chastellain, V.K. Parashar, A. Petri, H. Hofmann, M.A.M. Gijs, Plastic micropump with ferrofluidic actuation, J. Microelectromech. Syst., 14 (2005) 96-102.
[67] F.C.M. Vandepol, H.T.G. Vanlintel, M. Elwenspoek, J.H.J. Fluitman, A thermopneumatic micropump based on micro-engineering techniques, Sens. Actuator A-Phys., 21 (1990) 198-202.
[68] O.C. Jeong, S.W. Park, S.S. Yang, J.J. Pak, Fabrication of a peristaltic PDMS micropump, Sens. Actuator A-Phys., 123-24 (2005) 453-458.
[69] O.C. Jeong, S.S. Yang, Fabrication and test of a thermopneumatic micropump with a corrugated p plus diaphragm, Sens. Actuator A-Phys., 83 (2000) 249-255.
[70] J.H. Kim, K.H. Na, C.J. Kang, Y.S. Kim, A disposable thermopneumatic-actuated micropump stacked with PDMS layers and ITO-coated glass, Sens. Actuator A-Phys., 120 (2005) 365-369.
[71] C.Y. Lee, G.B. Lee, J.L. Lin, F.C. Huang, C.S. Liao, Integrated microfluidic systems for cell lysis, mixing/pumping and DNA amplification, J. Micromech. Microeng., 15 (2005) 1215-1223.
[72] J.F. Chen, M. Wabuyele, H.W. Chen, D. Patterson, M. Hupert, H. Shadpour, D. Nikitopoulos, S.A. Soper, Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate, Anal. Chem., 77 (2005) 658-666.
[73] G.Q. Hu, Q. Xiang, R. Fu, B. Xu, R. Venditti, D.Q. Li, Electrokinetically controlled real-time polymerase chain reaction in microchannel using Joule heating effect, Anal. Chim. Acta, 557 (2006) 146-151.
[74] E.T. Lagally, P.C. Simpson, R.A. Mathies, Monolithic integrated microfluidic DNA amplification and capillary electrophoresis analysis system, Sens. Actuators B-Chem., 63 (2000) 138-146.
[75] P.K. Yuen, L.J. Kricka, P. Wilding, Semi-disposable microvalves for use with microfabricated devices or microchips, J. Micromech. Microeng., 10 (2000) 401-409.
[76] Anderson RC, Su X, Bogdan GJ, Fenton J. A miniature integrated device for automated multistep genetic assays. Nucleic Acids Res., 28 (2000) e60.
[77] W.H. Grover, A.M. Skelley, C.N. Liu, E.T. Lagally, R.A. Mathies, Monolithic membrane valves and diaphragm pumps for practical large-scale integration into glass microfluidic devices, Sens. Actuators B-Chem., 89 (2003) 315-323.
[78] E.T. Lagally, C.A. Emrich, R.A. Mathies, Fully integrated PCR-capillary electrophoresis microsystem for DNA analysis, Lab Chip, 1 (2001) 102-107.
[79] E.T. Lagally, J.R. Scherer, R.G. Blazej, N.M. Toriello, B.A. Diep, M. Ramchandani, G.F. Sensabaugh, L.W. Riley, R.A. Mathies, Integrated portable genetic analysis microsystem for pathogen/infectious disease detection, Anal. Chem., 76 (2004) 3162-3170.
[80] N.M. Toriello, C.N. Liu, R.A. Mathies, Multichannel reverse transcription-polymerase chain reaction microdevice for rapid gene expression and biomarker analysis, Anal. Chem., 78 (2006) 7997-8003.
[81] C.N. Liu, N.M. Toriello, R.A. Mathies, Multichannel PCR-CE microdevice for genetic analysis, Anal. Chem., 78 (2006) 5474-5479.
[82] P. Liu, T.S. Seo, N. Beyor, K.J. Shin, J.R. Scherer, R.A. Mathies, Integrated portable polymerase chain reaction-capillary electrophoresis microsystem for rapid forensic short tandem repeat typing, Anal. Chem., 79 (2007) 1881-1889.
[83] C.S. Liao, G.B. Lee, H.S. Liu, T.M. Hsieh, C.H. Luo, Miniature RT-PCR system for diagnosis of RNA-based viruses, Nucleic Acids Res., 33 (2005).
[84] K.Y. Lien, W.C. Lee, H.Y. Lei, G.B. Lee, Integrated reverse transcription polymerase chain reaction systems for virus detection, Biosens. Bioelectron., 22 (2007) 1739-1748.
[85] F.C. Huang, C.S. Liao, G.B. Lee, An integrated microfluidic chip for DNA/RNA amplification, electrophoresis separation and on-line optical detection, Electrophoresis, 27 (2006) 3297-3305.
[86] M.A. Unger, H.P. Chou, T. Thorsen, A. Scherer, S.R. Quake, Monolithic microfabricated valves and pumps by multilayer soft lithography, Science, 288 (2000) 113-116.
[87] J. Liu, M. Enzelberger, S. Quake, A nanoliter rotary device for polymerase chain reaction, Electrophoresis, 23 (2002) 1531-1536.
[88] J. Liu, C. Hansen, S.R. Quake, Solving the "world-to-chip" interface problem with a microfluidic matrix, Anal. Chem., 75 (2003) 4718-4723.
[89] C.J. Packard, D.J. Rader, Lipids and atherosclerosis, Taylor & Francis, London, 2006.
[90] A.J. Lusis, Atherosclerosis, Nature, 407 (2000) 233-241.
[91] A.C. Newby, An overview of the vascular response to injury: a tribute to the late Russell Ross, Toxicol. Lett., 112 (2000) 519-529.
[92] R.O. Cannon, 3rd, Role of nitric oxide in cardiovascular disease: focus on the endothelium, Clin. Chem., 44 (1998) 1809-1819.
[93] S. Dimmeler, I. Fleming, B. Fisslthaler, C. Hermann, R. Busse, A.M. Zeiher, Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation, Nature, 399 (1999) 601-605.
[94] P. Libby, Vascullar biology of atherosclerolsis: Overview and state of the art, Am. J. Cardiol., 91 (2003) 3A-6A.
[95] N.A. Flavahan, Atherosclerosis or lipoprotein-induced endothelial dysfunction – potential mechanisms underlying reduction in edrf/nitric oxide activity, Circulation, 85 (1992) 1927-1938.
[96] A.D. Callow, Endothelial dysfunction in atherosclerosis, Vascul Pharmacol, 38 (2002) 257-258.
[97] R. Ross, Rous-Whipple Award Lecture. Atherosclerosis: a defense mechanism gone awry, Am. J. Pathol., 143 (1993) 987-1002.
[98] R. Ross, Atherosclerosis is an inflammatory disease, Am. Heart J., 138 (1999) S419-420.
[99] C. Monaco, E. Andreakos, S. Kiriakidis, M. Feldmann, E. Paleolog, T-cell-mediated signalling in immune, inflammatory and angiogenic processes: the cascade of events leading to inflammatory diseases, Curr. Drug Targets Inflamm. Allergy, 3 (2004) 35-42.
[100] R.M. Faruqi, P.E. Dicorleto, Mechanisms of monocyte recruitment and accumulation, British Heart Journal, 69 (1993) S19-S29.
[101] C.L. Klein, H. Kohler, F. Bittinger, M. Wagner, I. Hermanns, K. Grant, J.C. Lewis, C.J. Kirkpatrick, Comparative studies on vascular endothelium in vitro. I. Cytokine effects on the expression of adhesion molecules by human umbilical vein, saphenous vein and femoral artery endothelial cells, Pathobiology, 62 (1994) 199-208.
[102] R.J. Faull, Adhesion molecules in health and disease, Australian and New Zealand Journal of Medicine, 25 (1995) 720-730.
[103] P. Libby, P.M. Ridker, A. Maseri, Inflammation and atherosclerosis, Circulation, 105 (2002) 1135-1143.
[104] R.T. Dean, D.T. Kelly, Atherosclerosis : gene expression, cell interactions, and oxidation, Oxford University Press, Oxford, 2000.
[105] R. Ross, Atherosclerosis--an inflammatory disease, N. Engl. J. Med., 340 (1999) 115-126.
[106] R.R. Packard, P. Libby, Inflammation in atherosclerosis: from vascular biology to biomarker discovery and risk prediction, Clin. Chem., 54 (2008) 24-38.
[107] A. Zalewski, C. Macphee, Role of lipoprotein-associated phospholipase A(2) in atherosclerosis - Biology, epidemiology, and possible therapeutic target, Arterioscler. Thromb. Vasc. Biol., 25 (2005) 923-931.
[108] A.D. Tselepis, M. John Chapman, Inflammation, bioactive lipids and atherosclerosis: potential roles of a lipoprotein-associated phospholipase A2, platelet activating factor-acetylhydrolase, Atheroscler. Suppl., 3 (2002) 57-68.
[109] J.H. Min, M.K. Jain, C. Wilder, L. Paul, R. Apitz-Castro, D.C. Aspleaf, M.H. Gelb, Membrane-bound plasma platelet activating factor acetylhydrolase acts on substrate in the aqueous phase, Biochemistry, 38 (1999) 12935-12942.
[110] O.G. Berg, M.H. Gelb, M.D. Tsai, M.K. Jain, Interfacial enzymology: the secreted phospholipase A(2)-paradigm, Chem. Rev., 101 (2001) 2613-2654.
[111] R.A. Deems, Interfacial enzyme kinetics at the phospholipid/water interface: practical considerations, Anal. Biochem., 287 (2000) 1-16.
[112] V. Fuster, R. Ross, E.J. Topol, Atherosclerosis and coronary artery disease, Lippincott-Raven, Philadelphia, 1996.
[113] A.L. Lehninger, D.L. Nelson, M.M. Cox, Lehninger principles of biochemistry, W.H. Freeman, New York, 2005.
[114] J. Moiroux, P.J. Elving, Effects of adsorption, electrode material, and operational variables on oxidation of dihydronicotinamide asenine-dinucleotinamide at carbon electrodes, Anal. Chem., 50 (1978) 1056-1062.
[115] H. Jaegfeldt, Adsorption and electrochemical oxidation behavior of NADH at a clean platinum-electrode, J. Electroanal. Chem., 110 (1980) 295-302.
[116] S.A. Wring, J.P. Hart, Chemically modified carbon-based electrodes and their application as electrochemical sensors for the analysis of biologically important compounds – a review, Analyst, 117 (1992) 1215-1229.
[117] M. Hedenmo, A. Narvaez, E. Dominguez, I. Katakis, Reagentless amperometric glucose dehydrogenase biosensor based on electrocatalytic oxidation of NADH by osmium phenanthrolinedione mediator, Analyst, 121 (1996) 1891-1895.
[118] B.W. Carlson, L.L. Miller, Mechanism of the oxidation of NADH by quinones – energetic of one-electron and hydride routes, J. Am. Chem. Soc., 107 (1985) 479-485.
[119] A. Kitani, Y.H. So, L.L. Miller, An electrochemical study of the kinetics of NADH being oxidized by diimine derived form diaminobenzenes and diaminopyrimimids, J. Am. Chem. Soc., 103 (1981) 7636-7641.
[120] K. Essaadi, B. Keita, L. Nadjo, R. Contant, Oxidation of NADH by oxometalates, Journal of Electroanal. Chem., 367 (1994) 275-278.
[121] F.Q. Xu, H.W. Li, S.J. Cross, T.F. Guarr, Electrocatalytic oxidation of NADH at poly(metallopthalocyanine)-modified electrode, Journal of Electroanal. Chem., 368 (1994) 221-225.
[122] M. Ishiyama, K. Sasamoto, M. Shiga, Y. Ohkura, K. Ueno, K. Nishiyama, I. Taniguchi, Novel disulfonated tetrazolium salt that can be reduced to a water-soluble formazan and its application to the assay of lactate-deydrogenase, Analyst, 120 (1995) 113-116.
[123] C.E. Banks, R.G. Compton, Exploring the electrocatalytic sites of carbon nanotubes for NADH detection: an edge plane pyrolytic graphite electrode study, Analyst, 130 (2005) 1232-1239.
[124] J. Wang, G. Rivas, M. Chicharro, Glucose microsensor based on electrochemical deposition of iridium and glucose oxidase onto carbon fiber electrodes, Journal of Electroanal. Chem., 439 (1997) 55-61.
[125] M.C. Rodriguez, G.A. Rivas, Glucose biosensor prepared by the deposition of iridium and glucose oxidase on glassy carbon transducer, Electroanalysis, 11 (1999) 558-564.
[126] J. Shen, L. Dudik, C.C. Liu, An iridium nanoparticles dispersed carbon based thick film electrochemical biosensor and its application for a single use, disposable glucose biosensor, Sens. Actuators B-Chem., 125 (2007) 106-113.
[127] Y.C. Luo, J.S. Do, C.C. Liu, An amperometric uric acid biosensor based on modified Ir-C electrode, Biosens. Bioelectron., 22 (2006) 482-488.

校內：2011-08-15公開
校外：2018-08-15公開

簡易檢索 / 詳目顯示

相關論文