臺灣博碩士論文加值系統

第一部分
鮑氏不動桿菌為近年來嚴重之院內感染病原，尤其是多重抗藥性菌株的產生，造成治療上的困難。這些多重抗藥性菌株的治療需要使用到許多後線抗生素，老虎黴素即是其中之一。但隨著老虎黴素的使用，對他產生不具感受性的菌株也隨之出現。而這些抗藥性有絕大部分都與輸出幫浦AdeABC的表現增加有關，此輸出幫浦之調控是由上游的雙因子調控機制AdeR及AdeS所控制。利用電泳膠體遲滯的結果我們發現，調控因子AdeR會辨認到adeR與adeABC中間的順反子區間上的直接重複序列而非adeABC之啟動子，其結合會抑制下游輸出幫浦的表現。另外由臨床菌株發現在AdeR調控因子上核酸結合的功能區塊發生改變，會降低AdeR對於直接重複序列的結合能力，造成下游輸出幫浦表現量增加，對老虎黴素之抗藥性也增加 (MIC = 16 μg/mL)。此研究對輸出幫浦之調控機制探討能幫助我們更了解鮑氏不動桿菌產生抗藥性的原因，有助未來臨床治療及新藥開發之契機。
第二部分
本研究的目的在使用綠色合成的方式生產奈米銀。以硝酸銀為銀離子的前驅物，再以葡萄糖及三甲基硝酸幾丁聚醣(TMCN)分別作為還原劑以及穩定劑。在室溫下加入鹼性溶液後不斷攪拌即可完成奈米銀合成。過程中無需消耗能源或使用貴重儀器。調整氫氧化鈉、葡萄糖及三甲基硝酸幾丁聚醣的濃度會影響奈米銀之粒徑，表面電位及產量。所生產之奈米銀顆粒粒徑大小約60 nm，並帶有正電位。藉著針對三甲基硝酸幾丁聚醣修飾之奈米銀進行其物理及化學特性之鑑定，包括光子光譜儀鑑定、都卜勒雷射界面電位分析、 穿透式電子顯微鏡、X光繞射及X光電子能譜分析等。並以硼氫化鈉測定奈米銀之催化活性。藉由微量培養基稀釋法證明三甲基硝酸幾丁聚醣修飾之奈米銀對於鮑氏不動桿菌、大腸桿菌、綠膿桿菌、金黃色葡萄球菌皆具有抗菌活性。最小抑菌濃度(MIC)值小於6.13 μg/mL，另外此奈米銀對於多重抗藥性鮑氏不動桿菌也具有抗菌活性，其MIC值為小於 12.25 μg/mL。

Part 1
Acinetobacter baumannii is an increasing threat of nosocomial infections in recent years, especially the emergence of multi-drug resistant strains (multi-drug resistant Acinetobacter baumannii; MDRAB). Infections caused by multi-drug resistant A. baumannii could cause longer hospital stay and higher treatment costs. Tigecycline, a board-spectrum tetracycline derivative, is considered as the last antibiotic choice for the MDRAB. However, resistance to Tigecycline was reported following the drug usage worldwide. These resistances are mainly associated with overexpression of efflux pump, especially AdeABC, which is regulated by a corresponding two-component system AdeRS. In the research, we found that the regulatory factor AdeR could recognize the direct repeat on intercistronic region between adeR and adeA. This interaction inhibits the downstream efflux pump expression. In addition, mutations on AdeR DNA binding domain show lower affinity to the direct repeat sequences, and elevate the expression level of efflux pump, leading to high resistance to tigecycline (MIC = 16 μg/mL). This result is very useful for understanding the mechanism of tigecycline resistance of A. baumannii.
Part 2
We generate a silver nanoparticles using green synthesis. In this process, silver nitrate is used as a precursor of silver ions, and then glucose and trimethyl nitrate chitosan (TMCN) are used as a reducing agent and stabilizer, respectively. The whole reaction of silver nanoparticle synthesis could be done at room temperature after adding alkaline solution and mixing thoroughly. There is no need to consume energy or to use expensive equipment. Adjusting the concentration of sodium hydroxide, glucose and TMCN will affect the particle size, zeta potential and formation yield of silver nanoparticles. The average size of this silver nanoparticles (TMCN-AgNPs) is around 60 nm with positive surface charge. The physical and chemical properties of this nanoparticles were characterized by UV-Vis spectroscopy, X-ray diffraction and X-ray photoelectron spectroscopy. The catalytic activity of TMCN-AgNPs was determined by reduction of 4-nitrophenol using NaBH4 as a reducing agent. The antibacterial activity of TMCN-AgNPs was evaluated by broth microdilution method, and was proved to have antibacterial activity against Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The minimum inhibitory concentration (MIC) was < 6.13 μg/mL. Moreover, TMCN-AgNPs also showed antibacterial activity against multidrug-resistant Acinetobacter baumannii from clinical isolated, and the MIC value was < 12.25 μg/mL.

目錄
第一部分
鮑氏不動桿菌之AdeR調控蛋白對adeABC輸出幫浦表現之研究
摘要..........................................3
壹、研究目的...................................4
貳、材料與方法................................17
參、實驗結果..................................35
肆、討論 .....................................42
伍、圖表 .....................................49
陸、參考文獻..................................71

第二部分
具正介面電位三甲基硝酸幾丁聚醣修飾之奈米銀: 其催化特性及抗菌活性, 包括多重抗藥性鮑氏不動桿菌
摘要..........................................86
壹、研究目的...................................88
貳、材料與方法................................100
參、實驗結果..................................109
肆、討論......................................120
伍、圖表......................................127
陸、參考文獻..................................150

總結.........................................159

Part 1
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Part 2
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