1,721,069 research outputs found
The Functional Analysis of TIF1beta on the Regulation of Cell Cycle
No full textTIF1b 是一個上位遺傳調控因子，藉由與其它的染色質蛋白互相作用，能夠在基因體的特定區位上進行染色質的結構重整與轉錄調控。一般認為TIF1b 的抑制功能主要經由其與異染色質蛋白一號(Heterochromatin Protein 1, HP-1)的交互作用所達成。在TIF1b 上的HP-1 box 是主要負責這個交互作用的區段。然而，背後的調控機制仍未完全明瞭。在這篇論文中，我們建立了一個基因置換系統能夠將生性的蛋白質抑制，以外生性的突變蛋白質補回，藉以研究特定胺基酸位點對其功能之影響。此外，利用抗體的專一性，我們也發現TIF1b 能夠在HP-1 box 中的Ser473 上被磷酸化，而這樣的磷酸化是會隨著細胞週期而改變的。此外，利用染色質沉澱的技術，我們也觀察到這樣的磷酸化能夠確實影響到特定基因體區位上的HP-1 召集。最後，我們也觀察到了TIF1b 能夠與組蛋白乙烯基轉化脢PCAF形成錯合物，而此觀察也提供了TIF1b 做為活化因子的可能性。The transcriptional intermediary factor 1β (TIF1β)/KRIP-1/KAP-1/TRIM28 is an epigenetic regulator correlated to transcriptional regulation and chromatin remodeling at designated genomic loci through the interaction with other chromatinic proteins. The repressive capability of TIF1β is thought to be mediated in part by its interaction with HP1. The HP1-box, PXVXL, of TIF1β is reported to be responsible for this interaction.owever, the underlying regulation is still poorly understood. Here we demonstrate the construction of a gene replacement system which replaces the endogenous TIF1β withctopically expressed ones carrying desired alterations. Also, using phospho-specific antibody, it is shown that TIF1β is phosphorylated at Ser473, which is located in theP1-interacting domain, and the global level of this modification fluctuates with the progression of cell cycle. Furthermore, in our ChIP assay, it is shown that theverexpression of wild type and S473A but not the phospho-mimicry S473E mutant of TIF1β can preferentially enrich the recruitment of HP1β to endogenous promoter region of CDC2 and CDC25a. In conclusion, the phosphorylation of TIF1β Ser473 is demonstrated to be functionally correlated to the recruitment of HP1 protein to specific chromatinic loci. Finally, we’ve also provided the evidence that TIF1β can potentially complex with the histone acetyltransferases PCAF, and this interaction is possibly involved in the TIF1β-mediated transcriptional activation reported previously.致謝 3文摘要 5bstract 6aterial and Methods 10ell Culture 10lasmid Construction 10ransfection 11reparation of Whole Cell Extract 11ell Synchronization 11ntibody 12DS-PAGE and Western Blot 12hromatin Immunoprecipitation 13mmunostaining 14esult 15onstruction of Vector-based miRNA against mouse TIF1β 16IF1β Replacement 16he Dynamic Regulation of TIF1β Ser473 Phosphorylation during Cell Cycle Progression 16hospho-mimicry Mutation of TIF1β Ser473 Compromises the Recruitment of HP1β to E2F-responsive elements in vivo 17ndogenous TIF1β can be Co-immunoprecipitated by Overexpressed PCAF 18iscussion 20igure 23igure Legent 30eference 3
Generation and Analysis of Transgenic Mice and Pigs Harboring the Cellulase and Phytase Genes Drived by Porcine Pancreatic Amylase Promoter
No full text本研究旨在將分別由本土水牛瘤胃真菌及牛糞便中分離出之纖維素分解酶（cellulase）、植酸分解酶（phytase）基因與由豬胰臟選殖出之具組織特異性的胰澱粉酶基因啟動子（pancreatic amylase promoter）構築而成的轉殖基因（pAMY-CEL、pAMY-PHY），藉由基因顯微注射及胚移置的技術，產製攜帶有該基因之轉基因小鼠與轉基因豬，冀望達到改進家畜對飼料中纖維素與有機磷的消化與利用，並增加生長效率、減少排泄物，進而達到環保與提升農民收益的最終目標。
在轉基因小鼠產製之試驗中，合計完成267個小鼠原核胚之基因顯微注射，其中245個經基因注射後外表完整且初步判定存活之小鼠胚，分別移置於10隻受胚母鼠，待懷孕期滿後陸續產下合計63隻仔小鼠。離乳後僅存活57隻，分別剪取尾巴組織抽取基因組DNA（genomic DNA）進行聚合酶連鎖反應（polymerase chain reaction）及南方吸漬法（Southern blot）分析，證實其中4隻小鼠確係帶有pAMY-CEL基因者；有6隻小鼠確係帶有pAMY-PHY基因。待此等轉基因小鼠發育達性成熟後，除與具生育力之ICR雄性或雌性小鼠進行配種，以瞭解轉殖基因小鼠的性腺傳承外，並同時收集其排泄物，進一步分析其對纖維素與有機磷的消化利用情形。
在轉基因豬產製之試驗中，合計完成145個豬原核胚之基因注入，其中138個業經完成基因注射後外表完整且存活之豬胚，並移置於6頭發情同期化之受胚豬後，合計獲得13頭仔豬出生；經分別抽取其基因組DNA進行PCR反應及Southern blot分析結果，證實其中5頭(3♀ /2♂)仔豬確係帶有pAMY-CEL基因者；2頭(1♀ /1♂)仔豬確係帶有pAMY-PHY基因。其中除編號Tg 13-2雌性仔豬不幸於出生後第10日即告夭折外，目前均已發育迄保育階段，刻正期待達性成熟，並分別與生育力正常之非轉基因豬完成配種，俾進一步分析確認轉殖基因在性腺之傳承效率，及其對纖維素與有機磷的消化利用情形。Generation and Analysis of Transgenic Mice and Pigs Harboring the Cellulase and Phytase Genes Drived by Porcine Pancreatic Amylase Promoter
Yu-Sheng Lin
Abstract
The objective of this study was to generate transgenic animals carrying designed transgene(s) which includes a gastrointestinal track specific gene promoter and a structure genes derived from the fiber or phosphorous digestive enzymes of native water buffalo’s rumen microorganisms. Those transgenic animals are expected to have better ability to digest fibers, use the phosphorous more efficiently, produce less solid waste, and ultimately brought more profit to the farmers.
Base on the purpose described, we have cloned the promoter of porcine pancreatic amylase which would limit the secretion of target enzymes to the duodenum, avoiding the extreme pH value of stomach acid and retained the functions of proteins.
Using the technique of microinjection, the purified transgenes were delivered into the pronuclei of fertilized eggs that were surgically obtained from the donor animals. Those injected eggs then were transferred into the oviduct of the recipients.
To date, one hundred and forty five fertilized eggs were injected and transferred. There were two sows giving birth and thirteen piglets were born. Desired full-length transgenes were found in five of those piglets by the polymerase chain reaction (PCR) analysis. Further, the genomic DNA from these five piglets was extracted from ear pieces and was use in Southern blot analysis. Multiple bands were observed in most of the new born animals. It is possible that the transgenes were inserted as single copy and multiple copies which in the format of tail to tail or head to tail. Nonetheless, these results confirmed that piglets number 29-7 and 29-8 harboring transgenes, pAMY-PHY and pAMY-CEL; piglets number 29-3, 13-2, and 13-5 harboring single transgene pAMY-CEL.
For the experiments of producing transgenic mice, there are sixty three pups were born and genomic DNA was extracted from their tails and used in PCR analysis. There are four pups were found carrying the desired transgenes in full length. The Southern blot analysis is currently on going.目錄
目錄……………………………………………………………….……………..I
表次…………………………………………………………………………....III
圖次………………………………………………………………...………….IV
摘要……………………………………………………………….………..……1
前言………………………………………………………………………...……2
壹、文獻檢討
一、豬之消化道生理………………...…………………………………..….4
二、胰臟……………………..………………………………………..……..5
三、植酸分解酶……………………..………………………………………9
四、纖維素分解酶………..………………………………………..............12
五、產製轉基因動物之方法………….……………………………………14
六、產製轉基因動物之目的………………………………………………19
貳、材料與方法
一、分子選殖之方法………………………………………………………..23
二、轉殖基因之構築……………………………..…….………………......26
三、試驗動物之飼養及超級排卵處理………………..…………………....37
四、基因轉殖動物之分析………………………………..………………..41
參、結果與討論
一、豬胰澱粉酶啟動子之分子選殖…………………………….…………46
二、攜帶pAMY-CEL、pAMY-PHY轉基因小鼠之產製與分析…………50
三、攜帶pAMY-CEL、pAMY-PHY轉基因豬之產製與分析……………55
結論…………………………………………………………………………63
參考文獻…………………………………………………………………….64
英文摘要…………………………………………………………..……….74
作者小傳……………………………………………………………….…76
表 次
表 1.利用GenomeWalker 工具 方法中所使用之寡核苷酸引子……….34
表 2.轉殖基因1216AMY-phrGFP pAMY-CEL和pAMY-PHY基因構築所設計的引子.……………………………….…………..……..…………..36
表 3.偵測帶有pAMY-CEL和pAMY-PHY基因之基因轉殖小鼠及豬時所使用之寡核苷酸引子配對……..……………………………………44
表 4.以基因顯微注射法產製pAMY-CEL和pAMY-PHY 基因轉殖小鼠之效率…………………………………………….………….…………54
表 5.以基因顯微注射法產製pAMY-CEL和pAMY-PHY 基因轉殖豬之效率………………………………………………………………………61
圖 次
圖1.豬隻消化道簡圖………..……………………………………….………6
圖2. α-澱粉酶之立體結構…………………………………………………. 8
圖3植酸之構造…………………………………………………………….11
圖4.纖維素之構造…………………………………..………………….......13
圖5.纖維素分解酶基因主要選殖於瘤胃真菌…………………………….27
圖6.植酸分解酶基因主要選殖於牛糞便中的大腸桿菌…………………28
圖7. PCR選殖載體系統。分別將PCR產物進行接合反應進入pGEM®-T Easy vector中，轉形成質體，並進行定序確定基因序列.…………29
圖8.轉基因之構築策略。將先將選殖出來的胰臟澱粉酶啟動子基因利用接合反應構築在此表現質體上，再將結構基因接合上，形成完整之transgenes（pAMY-CEL and pAMY-PHY）...…………...……………30
圖9.哺乳動物細胞中表現綠色螢光蛋白的表現載體之構築。本研究將豬胰澱粉酶啟動子置入此載體中，並轉染於胰臟腫瘤細胞（AR -42J），用來證明啟動子的活性…………………………….……….31
圖10. GSP1、GSP2、GSP3在豬隻胰臟澱粉酶基因序列上之位置…….32
圖11. genome walkerTM kit實驗方法之簡單意示圖..............…………….35
圖12. 供胚鼠發情同期化暨超級排卵流程圖…………….……………... 38
圖13.供胚豬與受胚豬發情同期化暨超級排卵流程圖.. …....……………40
圖14. 1216pAMY- phrGFP在螢光顯微鏡下（200X）所表現的綠色螢光。A為亮視野，B為暗視野.…………………………….……….……48
圖15. 412pAMY-phrGFP在螢光顯微鏡下（200X）所表現的綠色螢光。A為亮視野，B為暗視野.…..……………………….………………..49
圖16.應用聚合酵素連鎖反應技術結果證實攜帶有pAMY-CEL基因之轉基小鼠……………..………………………..…..………….………..52
圖17.應用聚合酵素連鎖反應技術結果證實攜帶有pAMY-PHY基因之轉基小鼠.………..…..……………..……………………………………..53
圖18.應用聚合酵素連鎖反應技術結果證實攜帶有pAMY-CEL基因之轉基因豬..….………..……………………....…………….………….... 57
圖19. 應用聚合酵素連鎖反應技術分析結果證實帶有pAMY-PHY基因之轉基因豬.…………………………………..…………...…………..58
圖20. 應用南方吸漬法分析結果證明帶有pAMY-CEL基因之轉基因豬.………….………………..………………………………………59
圖21. 應用南方吸漬法分析結果證明帶有pAMY-PHY 基因之轉基因豬.………………………….…………………....…………..……....60
圖22. 攜帶pAMY-CEL、pAMY-PHY基因之五頭轉基因豬……………. 6
Utilization of pork hydrolysate to prepare meat flavoring and studies on the formation of meat aroma
No full text本研究使用五種不同的商用蛋白酶(Bromelain、Flavourzyme、Neutrase、Papain、Alcalase)對碎豬肉進行水解。尋求對豬肉水解效果最佳的酵素，並建立最適的水解條件(溫度、振盪混合速率、水解時間)。結果顯示，以Flavourzyme對碎豬肉進行水解有最好的水解效率。以2% Flavourzyme在40℃、150 rpm下水解10小時為最佳的水解條件，所獲得的水解液之水解率平均為48.8 %。用所製備的豬肉蛋白水解液與其它肉香前驅物(胺基酸、還原糖、核苷酸等)進行熱反應來生產肉類香味料。水解液依序添加不同含量的cysteine、glycine、ribose、thiamine 及 IMP和GMP等物質進行熱反應(pH 7，121℃、30分鐘)並進行感官品評，尋求最佳的反應配方及添加量。再使用最佳反應配方在不同pH下(pH 5~9)進行熱反應並進行官能品評，求出最適的反應pH條件。結果顯示，在10 mL水解液中添加0.25 g cysteine、0.15 g glycine、0.5 g ribose、0.15 g thiamine和0.15 g IMP和GMP，在pH 6，121℃下反應30分鐘所生產的肉類香味料，其品評的分數最高(7.33)，具有接受性良好的肉類香味。pH 5~9的肉類香味料中一共鑑定出60種的香氣化合物，其中以雜環類化合物的種類(40種)及含量最多。在40種的雜環類化合物中有23種為含硫(S)化物。pH 5的部份產生的風味化合物較少。pH 5~9中，pyrazine的含量有隨著pH的上升而逐漸增加的趨勢。分析中鑑定出含量最多的化合物為4-methyl-5-thiazoleathanol，佔總量的50 %以上。該化合物的閾值偏高，其值為10.8 ppm，並非關鍵的肉類香氣化合物。本實驗只有在pH 6樣品中鑑定到2-methyl-3-furanthiol，其含量約佔總量的0.11 ％，為重要的肉類香味化合物。In this study, five different commercial proteases (Bromelain, Flavourzyme, Neutrase, Papain and Alcalase) was used to hydrolyze ground prok in order to find the protease which had the highest pork hydrolytic efficiency and established the optimal hydrolytic conditions (temperature, rotate speed and time). The result indicated that utilization of Flavourzyme to hydrolyze pork had the highest hydrolytic efficiency. The optimal hydrolytic condition is using 2 % Flavourzyme to hydrolyze pork for 10 hours at 40℃ and 150 rpm. The degree of hydrolysis of hydrolysate is 48.8 %. ork protein hydrolysate reacted with meat aroma precursors (amino acids, reducing sugars and nucleotides) to produce meat flavoring by thermal reaction. Hydrolysate reacted (pH 7, 121℃ for 30 min) with different content of cysteine, glycine, ribose, thiamine and IMP&GMP, the samples were tasted and scored by scorer. The best reaction species and contents were found. Reactions were carried out with different pH conditions (pH 5, 6, 7, 8, 9) by thermal reaction. The result showed that 10 mL hydrolysate reacted (pH 6, 121℃ for 30 min) with 0.25 g cysteine、0.15 g glycine、0.5 g ribose、0.15 g thiamine and 0.15 g IMP&GMP to produce the meat flavoring which had the highest score (7.33) and flavorable meat aroma. ixty compounds were identified from different pH (pH5~9) meat flavoring. There were fourty heterocyclic compounds in total meat flavoring. The content of heterocyclic compounds in meat flavoring was more than other compounds. There were twenty-three sulfides in forty heterocyclic compounds. The pH 5 meat flavoring had fewer flavor compounds. In all pH samples, when pH was increasing, pyrazine content was increasing too. 4-methyl-5-thiazoleathanol was identified from all pH samples which had the highest content (above 50 %). The threshold value of 4-methyl-5-thiazoleathanol is 10.8 ppm. It is not the critical compound of meat aroma. In this study, 2-methyl-3-furanthiol was only identified from pH 6 sample. The content of 2-methyl-3-furanthiol is 0.11 %. It is the important compound of meat aroma.口試委員會審定書...........................................i謝......................................................ii文摘要.................................................iii文摘要..................................................iv一章、研究動機...........................................1二章、文獻整理...........................................2.1食品風味................................................2.2.1 滋味化合物...........................................2.1.2 香味化合物...........................................2.1.3 風味增強劑...........................................3.2肉類香味................................................3.2.1 肉類香味料...........................................4.2.2 蛋白質水解物.........................................5.2.3 豬肉不同部位的選擇對蛋白水解物之影響.................6.3 肉類香味前驅物之探討...................................8.3.1 肉中之水溶性部分含有肉香的前驅物.....................8.3.2 胺基酸和醣類為生成共通肉類香氣的前驅物...............8.3.3 油脂為肉類特徵氣味的前驅物...........................8.3.4 三甘油酯並非肉香的前驅物.............................9.3.5 磷脂質為肉香的前驅物.................................9.3.6 維生素E對肉類香味的影響..............................9.3.7 核苷酸對肉類香氣的影響..............................10.3.8 維生素B1(硫胺素)對肉類香氣的影響....................10.4 肉類揮發性香氣的生成反應..............................13.4.1 梅納反應............................................13.4.1.1 梅納反應的機制....................................13.4.1.2 影響梅納反應的因素................................14.4.2 油脂的氧化..........................................16.4.3 維生素B1(硫胺素)的降解..............................17.4.4 蛋白質和游離胺基酸的熱降解..........................17.4.5 醣類的降解..........................................18.4.6 不同反應途徑間的交互作用............................18.5 肉類的香氣化合物......................................22三章、材料與方法........................................24.1 實驗架構..............................................24.1.1 豬肉之最適水解條件探討..............................24.1.2 肉類香味料反應配方與pH條件之探討....................24.1.3 肉類香味料熱反應後的ribose損失率和揮發性成分鑑定....25.2 實驗材料與器材........................................26.2.1 材料................................................26.2.2 試藥................................................26.3 實驗方法..............................................27.3.1 豬肉一般成分分析....................................27.3.1.1 水分含量測定......................................27.3.1.2 灰分含量測定......................................27.3.1.3 粗脂肪含量測定....................................27.3.1.4 粗蛋白含量測定....................................28.3.1.5 粗纖維含量測定....................................28.3.2 豬肉之最適水解條件探討..............................29.3.2.1 蛋白酶和熱處理對碎豬肉之水解率的影響..............29.3.2.2 溫度對碎豬肉之水解率的影響........................29.3.2.3 振盪混合速率對碎豬肉之水解率的影響................29.3.2.4 蛋白酶添加量及時間對碎豬肉之水解率的影響..........29.3.3 水解液之水解率相關測定..............................30.3.3.1 總氮 ( Total nitrogen, TN ).......................30.3.3.2 甲醛態氮 ( Formol nitrogen, FN )..................30.3.3.3 蛋白質水解率 (Degree of hydrolysis, DH, FN/TN)....31.3.4 肉類香味料反應配方與pH條件之探討....................31.3.4.1 Cysteine和glycine之添加量對肉類香味料的影響.......31.3.4.2不同醣類之添加對肉類香味料的影響...................31.3.4.3 Ribose之添加量對肉類香味料的影響..................31.3.4.4 Thiamine之添加量對肉類香味料的影響................31.3.4.5 IMP和GMP之添加量對肉類香味料的影響................32.3.4.6反應pH對肉類香味料的影響...........................32.3.4.7 肉類香味料之感官品評..............................32.3.5 肉類香味料熱反應後的ribose損失率....................33.3.6 肉類香味料之揮發性成分鑑定..........................33四章、結果與討論..................................34.1 豬後腿瘦肉之一般成分分析..............................34.2 豬肉之最適水解條件探討................................36.2.1 蛋白酶和熱處理對碎豬肉之水解率的影響................36.2.2 溫度對碎豬肉之水解率的影響..........................40.2.3 振盪混合速率對碎豬肉之水解率的影響..................40.2.4 水解時間對碎豬肉之水解率及水解液氣味的影響..........43.3 肉類香味料反應配方與pH條件之探討......................46.3.1 Cysteine和glycine的添加量對肉類香味料的影響.........46.3.2不同醣類之添加對肉類香味料的影響.....................46.3.3 Ribose之添加量對肉類香味料的影響....................50.3.4 Thiamine之添加量對肉類香味料的影響..................50.3.5 IMP和GMP之添加量對肉類香味料的影響..................53.3.6 反應pH對肉類香味料的影響............................53.4 肉類香味料熱反應後的ribose損失率......................56.5 肉類香味料之揮發性成分鑑定............................59五章、結論..............................................71六章、參考文獻..........................................73七章、附錄..............................................79目錄2-1、核糖核苷酸可轉換為肉類香氣化合物...................112-2、維生素B1的熱降解...................................122-3、半胱胺酸的熱降解可產生硫化氫、乙醛、氨和二氧化碳...224-1、未經加熱之碎豬肉以不同蛋白酶水解之水解率...........394-2、經預加熱之碎豬肉以不同蛋白酶水解之水解率...........394-3、使用不同醣類製備之樣品的顏色.......................494-4、Ribose標準品的液相層析圖譜.........................564-5、(A) pH 6樣品熱反應前的液相層析圖譜.................574-5、(B) pH 6樣品熱反應後的液相層析圖譜.................574-6、pH 6樣品之二氯甲烷萃出物中的揮發性成分之氣相層析圖.60目錄2-1、豬肉不同部位之一般組成成分..........................72-2、維生素B1在不同系統模式下所產生的熱降解產物.........202-3、維生素B1和半胱胺酸反應所產生的熱降解產物...........212-4、熟牛肉中的重要肉類香氣化合物.......................234-1、本實驗之豬後腿瘦肉一般成分分析.....................354-2、未經加熱之碎豬肉以不同蛋白酶水解之水解率...........374-3、經預加熱之碎豬肉以不同蛋白酶水解之水解率...........384-4、溫度對碎豬肉之水解率的影響.........................414-5、振盪混合速率對碎豬肉之水解率的影響.................424-6、水解時間對碎豬肉之水解率及水解液氣味的影響.........444-7、蛋白酶添加量及水解時間對碎豬肉之水解率的影響.......454-8、不同cysteine及glycine添加量對肉類香味料之香氣接受性喜好度的影響........474-9、不同醣類對肉類香味料之香氣接受性喜好度的影響.......484-10、不同ribose添加量對肉類香味料之香氣接受性喜好度的影響................514-11、不同thiamine添加量對肉類香味料之香氣接受性喜好度的影響................524-12、不同IMP和GMP添加量對肉類香味料之香氣接受性喜好度的影響................544-13、pH對肉類香味料之香氣接受性喜好度的影響............554-14、不同pH樣品熱反應後的ribose損失率..................584-15、pH 5樣品中所鑑定出的香氣化合物....................614-16、pH 6樣品中所鑑定出的香氣化合物....................624-17、pH 7樣品中所鑑定出的香氣化合物....................634-18、pH 8樣品中所鑑定出的香氣化合物....................644-19、pH 9樣品中所鑑定出的香氣化合物....................654-20、pH 5~9樣品中所鑑定出的香氣化合物之比較...........6
Sex, Menopause, Metabolic Syndrome, and All-Cause and Cause-Specific Mortality-Cohort Analysis from the Third National Health and Nutrition Examination Survey
No full textObjective: This study assessed the mortality risk associated with metabolic syndrome (MetS) for participants from the Third National Health and Nutrition Examination Survey. Design, Setting, and Patients: The study analyzed mortality data from 1364 men and 1321 women aged 40 yr and older based on their MetS status defined by National Cholesterol Education Program Adult Treatment Panel III. Subjects initially using insulin, oral hypoglycemic, antihypertensive , or lipid-lowering medications were excluded. Main Outcome Measures: All-cause, cardiovascular, cardiac, and noncardiovascular mortality were obtained from the Third National Health and Nutrition Examination Survey-linked mortality follow-up file through December 31, 2000. Results: The prevalence of MetS was 33 and 29% for men and women, respectively. In the male subjects, there was no significant association between MetS and mortality. In the women, MetS was an independent risk factor for all-cause mortality [ hazard ratio (HR) 1.84, 95% confidence interval (CI) 1.29-2. 64, P = 0.001], cardiovascular mortality (HR 1.96, 95% CI 1. 21-3.17, P = 0.007), cardiac mortality (HR 1 .88, 95% CI 1.15 -3.09, P = 0.01), and noncardiovascular mortality (HR 1. 80, 95% CI 1.13-2.87, P = 0.01). The HR was stronger when postmenopausal women were analyzed separately and became nonsignificant in the premenopausal cohort. The sex-specific HR remained unchanged, regardless of the MetS criteria used or the inclusion of actively treated subjects. Conclusions: MetS poses a significant increase in mortality risk through an observation period as long as 12 yr, primarily in postmenopausal women, that is not apparent in men and premenopausal women. Sex is an important effect modifier of all-cause and cause-specific death
Metabolic Syndrome and Long-Term Outcomes in United States Adults - Cohort Analysis from the Third National Health and Nutrition Survey (Nhanes Iii)
No full textModal sensitivities, spatial signal distribution andverage of segmented ring sensors and control effect ofegmented ring actuators
No full text對於薄殼類型之結構的分佈控制，必須要有效的致動和動態析。本研究是估算片段分佈的電-機械性質的壓電薄殼感測器用在環薄殼。由於感測器的訊號是模態決定，所以必須先學習然模態的特性而且特別強調於環向之模態。其次，在分佈片段感測器使用分割的技巧且設計不同的長度，使得藉著模態形變估算訊號。此，全部的輸出訊號總共包含了四個部份：1)環向之振動振幅所造成彎曲應變而感生訊號，2)徑向之振動的振幅所造成曲應變而感生訊號，3)環向之振動的振幅所造成膜應變而感生號，4)徑向之振動的振幅所造成膜應變而感生訊號。膜應變與曲應變主要分別由環向與徑向之自然頻率伴隨而生。所以感測的敏感度可以在分成：1)徑向型態敏感度與2)環向型態敏感。用參數學習(例如：環的半徑、感測器的厚度、環的厚度片段感測器的大小)而引導計算出在自然振動之下，所產生在的壓電片感測器所得之訊號。所有的資料指示著，產生的總訊主要是由環向分量貢獻為較多，而不是徑向分量。次，在環薄殼上包覆致動層，藉著推導致動片之控制效，瞭解在徑向模式時，彎曲控制效應支配整個控制效應，反之環向模式時，膜控制效應會是主要控制效應的主幹。後，使用參數學習（例如：環的半徑、致動器的厚度、環厚度和片段致動器的大小與施加電壓大小）而瞭解這些幾何參影響控制效應之大小。Distributed monitoring is essential to effective actuation andibration control of shell-type structures. In this thesis, electro-mechanicsf the segmented distributed piezoelectric shell sensors applied to ringhells are evaluated. Since sensor signals are modal dependent, naturalodal characteristics, emphasizing the circumferential mode, are studiedirst. Then, with the segmentation technique, distributed segmentedensors with various lengths are designed and their signal generationsesulting from modal strains are evaluated.he total output signal includes four components: 1) a bending strainnduced signal by the circumferential oscillatory amplitude, 2) a bendingtrain induced signal by the transverse oscillatory amplitude, 3) aembrane strain induced signal by the circumferential oscillatorymplitude and 4) a membrane strain induced signal by the transversescillatory amplitude respectively at the transverse and circumferentialomponent frequencies. Furthermore, the sensor sensitivities are dividednto: 1) the transverse modal sensitivity and 2) the circumferential modalensitivity.arametric studies (e.g., ring radius, sensor thickness, ring thicknessnd sensor segment length) are conducted to evaluate the spatial signalistributions and component signal generations of segmented ring sensors.ll data indicate that ring’s circumferential, not transverse, componentode dominates the total signal generation.hen, distributed control actions induced by electrostrictive actuatoregments are evaluated the total effect can be divided into twoicroscopic control actions: the circumferential membrane and bendingontrol actions. The bending control action in total control action is majorontribution at the transverse mode. The membrane control action in totalIontrol actions is major contribution at the circumferential mode.inally, parametric studies (e.g., ring radius, actuator thickness, ringhickness, actuator segment length and applied voltage) are conducted tovaluate control effect.目錄試委員審定書........................................................................................ I謝............................................................................................................II要.......................................................................................................... IIIBSTRACT..............................................................................................V錄.........................................................................................................VII目錄...................................................................................................... IX目錄................................................................................................... XIV1 章 序論..............................................................................................1.1 引言..................................................................................................1.2 介紹..................................................................................................4.3 研究範圍..........................................................................................52 章 環之結構與震動分析理論..........................................................7.1 樂夫(LOVE)方程式..........................................................................7.2 深薄殼方程式................................................................................14.2.1 膜作用力(Membrane Forces).................................................15.2.2 彎曲力距(Bending Moments)................................................16.3 環薄殼運動方程式........................................................................16.4 環薄殼之自然振動分析................................................................183 章 環之感測器................................................................................21.1 感測器之輸出訊號........................................................................21.2 感測器之分割技術........................................................................24.3 感測器之敏感度............................................................................274 章 環之致動器................................................................................29.1 分佈電滯性致動片之控制力........................................................30.2 分佈致動器控制效應....................................................................32.3 環薄殼之致動器............................................................................36.4 致動器之效應................................................................................395 章 結果與討論................................................................................42.1 自然頻率與振幅比........................................................................45.2 模態型式........................................................................................48.3 環之壓電片感測器的輸出訊號....................................................57.4 敏感度分析....................................................................................72.5 環之電滯性致動器控制效應........................................................826 章 總結..........................................................................................10
Dissection of the Cryptococcus neoformans light response and characterization of Cryptococcus neoformans SSN8 homologue
No full text隱球菌為人體重要病原真菌。實驗室過去的研究發現，藍光抑制隱球菌生殖菌絲的形成。進一步利用同源序列的比對，在隱球菌基因體中，找到真菌保守性藍光調控基因CWC1及CWC2，並證明二者為隱球菌藍光反應不可或缺的因子(Lu et al., 2005)。為進一步探討隱球菌光反應Cwc複合體下游抑制菌絲形成的相關機制，本研究以農桿菌逢機突變轉殖技術，利用 CWC1基因過度表現株完全抑制生殖菌絲形成的特性，進行回復生殖菌絲生長突變株之篩選。利用此suppressor screening的策略，經篩選4132個農桿菌轉殖株，其中64株具有不同程度回復生殖菌絲生長之情形。其中一農桿菌轉殖株AZ5，T-DNA插入破壞CWC2基因之表現，印證了本篩選策略的有效性。另一轉殖株AY18，經實驗證明，T-DNA插入破壞隱球菌SSN8同源基因，並因此造成生殖菌絲回復生長之特徵。啤酒酵母菌SSN8基因，又稱為SRB11/UME3/CYCC/RYE2，為轉錄調控mediator complex的一員，參與糖類利用、減數分裂、細胞生長，以及轉錄反應等生理之調控。為瞭解SSN8基因在隱球菌中所扮演之角色，本研究亦建構野生型ssn8突變株、回復突變株，及過度表現株。SSN8基因突變造成的性狀包括，生殖菌絲的減少， haploid fruiting菌絲的大量生成、黑色素的明顯累積，而在營養充分的培養條件下，ssn8突變株細胞形態發生改變，並且在固體培養基表面可見侵入菌絲的形成等。相關菌株性狀分析的結果顯示，SSN8基因參與隱球菌許多生理及發生的過程，並且為負向調控之角色，而SSN8基因如何調控相關生理反應，以及其與Cwc複合體間的關連性，有待進一步的探討。Cryptococcus neoformans is an important human fungal pathogen. In our previous investigation, we demonstrated that blue light negatively regulates the sexual filamentous growth via the Cwc1/Cwc2 complex in C. neoformans. To further understand how light inhibits mating filamentation, we conducted a suppressor screening using Agrobacterium-mediated insertional mutagenesis to identify the components interacting with or downstream of the Cwc complex under the CWC1 overexpression strain background. Among 4132 nourseothricin-resistant transformans screened, we identified 64 strains restored different levels of mating filamentation. The sites of T-DNA integration among 21 transformans were recovered. One strain, AZ5, restored the highest level of filamentation was observed and that T-DNA was found to be inserted at the 5’-UTR region of the CWC2 gene. The finding of CWC2 gene in the screen validates our approach. In this study, we further characterized the transformant AY18 in which T-DNA integration affected the activity of a gene homologous to the Saccharomyces cerevisiae SSN8. In budding yeast, SSN8 gene, also named SRB11/UME3/CYCC/RYE2, has been demonstrated to play diverse roles in sugar utilization, meiosis, cell phase control, and transcription regulation. To reveal the roles of SSN8 gene in C. neoformans, we generated the ssn8 mtant, reconstituted strain, and overexpression strain under the wild-type background. The mutant phenotypes included slightly reduced mating filamentation, extensive haploid filamentation, increased melanin accumulation, changed cellular morphology and production of invasive hyphae under rich YPD growth condition. Our results suggest that C. neoformans SSN8 homologue plays critical roles in the diverse physiological and differentiation processes. This is the first demonstration of mediator component in C. neoformans. Its roles related to the Cwc complex remain to be further investigated.中文摘要 1
英文摘要 2
第一章 研究動機 4
第二章 前人研究 5
2.1隱球菌 (Cryptococcus neoformans) 5
2.2隱球菌生活史 6
2.3隱球菌致病因子 7
2.4 隱球菌光反應之研究 9
2.5 隱球菌轉殖方法之研究 11
2.6啤酒酵母菌SSN8同源基因之研究 13
2.6.1 SSN8同源基因的發現 14
2.6.2 UME gene family的發現 15
2.6.3 RYE gene family的研究 17
2.6.4 C-type cyclin之研究 18
2.6.5 The mediator complex 20
2.7 SSN3同源基因之研究 23
第三章 材料與方法 25
3.1 實驗菌株以及培養條件 25
3.2農桿菌基因轉殖流程 25
3.2.1 農桿菌以及隱球菌的培養 26
3.2.2 農桿菌的活化 26
3.2.3 ATMT感染試驗 26
3.2.4轉殖株的篩選 27
3.3 農桿菌轉植菌落的篩選 27
3.4 T-DNA插入位點目標基因之鑑定 28
3.5 SSN8同源基因突變質體之建構及SSN8同源基因突變株之篩選 29
3.6 SSN8基因回覆突變株質體的製備及回覆突變株的篩選 30
3.7 SSN8基因過度表現株的製備以及篩選 30
3.8基因槍轉殖技術 (biolistic transformation) 30
3.8.1 宿主細胞的培養 31
3.8.2宿主細胞的製備 31
3.8.3轉殖DNA的製備 32
3.8.4基因槍之操作流程 32
3.9隱球菌少量DNA的抽取 33
3.10隱球菌大量DNA之萃取 34
3.11隱球菌RNA的抽取 34
3.12南方雜合分析 35
3.13北方雜合分析 37
3.14菌株交配子代之分離實驗 38
3.15交配分析試驗(Mating assay) 39
3.16單核菌絲之分析實驗 ( Monokaryotic fruiting assay ) 39
3.17黑色素形成分析 (Melanin assay) 39
3.18莢膜表現性狀之分析 40
3.19親緣樹狀分析 (Phylogenetic analysis) 40
第四章 結果 41
4.1農桿菌轉殖菌株之篩選 41
4.2 T-DNA插入破壞目標基因之分析 42
4.3隱球菌農桿菌突變轉殖菌株AZ5之分析 43
4.4隱球菌農桿菌突變轉殖株AY18菌株表現型態及T-DNA插入分析 44
4.5隱球菌CWC1基因過度表現之ssn8突變株之建構 46
4.6隱球菌CWC1基因過度表現ssn8突變株之表現型態分析 46
4.7隱球菌SSN8同源基因之探討 46
4.8 ssn8野生型突變株、回復突變株及SSN8基因過度表現株之建構 48
4.8.1 ssn8野生型突變株之篩選 48
4.8.2 ssn8 + SSN8回覆突變株的建構 48
4.8.3 SSN8同源基因過度表現株的建構 49
4.9 ssn8突變株與SSN8基因過度表現菌株之表現型態分析 49
4.9.1生殖菌絲的形成分析 49
4.9.2 Haploid fruiting 單核菌絲的形成分析 50
4.9.3黑色素的形成分析 51
4.9.4 37℃生長測定 52
4.9.5菌株細胞型態之觀察 52
4.9.6侵入菌絲之形成觀察 52
4.10 ssn8 cwc1及ssn8 ste12α等雙突變株表現型態之觀察 53
第五章 討論 55
圖表 65
參考文獻 106
附錄 118
表目錄
表一、本實驗所使用之隱球菌菌株 66
表二、本實驗所使用之引子 69
表三、隱球菌農桿菌突變轉殖株 70
表四、定序確認插入位置之隱球菌農桿菌突變轉殖菌株 71
圖目錄
圖一、隱球菌農桿菌突變株生殖菌絲形成及南方雜合分析 73
圖二、隱球菌農桿菌突變轉殖株AZ5之生殖菌絲表現型態與T-DNA插入位置之分析 75
圖三、隱球菌農桿菌突變轉殖株AY18之表現型態分析 77
圖四、隱球菌農桿菌轉殖株AY18 T-DNA插入位置之確認 79
圖五、隱球菌農桿菌轉殖株AY18子代之基因型分析 81
圖六、隱球菌Ssn8與其他同源蛋白質胺基酸序列之比對及親源分析結果 83
圖七、隱球菌SSN8基因突變載體之建構及YKC38突變轉殖株之南方雜合分析結果 85
圖八、有性生殖子代PCR篩選確認各種基因型之ssn8突變株 87
圖九、隱球菌SSN8基因突變載體及各式ssn8突變株與回覆突變株之南方雜合分析結果 89
圖十、隱球菌ssn8突變株、ssn8 + SSN8回覆突變株，及SSN8基因過度表現株之生殖菌絲形成分析 91
圖十一、隱球菌ssn8突變株，ssn8 + SSN8回覆突變株，以及SSN8基因過度表現株monokaryotic fruiting菌絲之形成分析 93
圖十二、隱球菌ssn8突變株，ssn8 + SSN8回覆突變株，以及SSN8基因過度表現株之黑色素生成分析 95
圖十三、隱球菌ssn8突變株，ssn8 + SSN8回覆突變株，以及SSN8基因過度表現株之37℃生長測定 97
圖十四、隱球菌ssn8突變株，ssn8 + SSN8回覆突變株，以及SSN8基因過度表現株細胞形態之觀察99
圖十五、隱球菌ssn8突變株侵入菌絲之觀察101
圖十六、cwc1 ssn8與ste12α ssn8雙突變株生殖菌絲形成之觀察103
圖十七、cwc1 ssn8與ste12α ssn8雙突變株單核菌絲形成之觀察10
Platform of Personal Health Records for Diabetes Mellitus Care
No full text慢性疾病的照護需要病人自我健康管理的配合，藉由改善病人的生活習慣以達到緩和或改進病人的疾病與健康情況。 傳統的自我健康管理，病人利用紙本或是文書軟體紀錄自己的生活習慣、生理數值、或是用藥以及就醫紀錄等。此種方式在資料管理及維護上有諸多不便之處。本篇研究希望藉由實行個人健康管理平台，幫助病人紀錄生理數值和生活習慣與個管師良好互動，最後達到促進病人主動積極的參與健康照護並改善其健康狀況。The health care of chronic disease need combine with patient self-management for improving patient''s lifestyle and health behavior and finally improving patient''s disease and health status. In traditional self-management, patients record their health behavior, vital signs, medication and healthcare encounters on paper or by word processing software. These recording manners may result in difficult to maintain or manage data. This study was to build a personal health record application to help patients record their vital signs, health behavior and message or interact with case managers, and finally assist patients in actively involving their health care and improving their health status
Development of an Intelligent Scheduling System for a Semiconductor Foundry Fab
No full text由於晶圓廠就如同一個大型的彈性製造系統且晶圓的製程相當複雜，因此派工問題對於生產控制扮演相當重要的角色。對於動態的派工系統而言有兩個關鍵性的議題主宰著彈性製造系統的效能；其中一個就是如何選出適當且關鍵的顯著系統資訊當作判斷系統狀況的指標，另外一個就是派工系統的設計方法-也就是所謂分類機器的設計。相對於單一個派工法則，如果我們能有效的依據一些有用的系統的資訊為當時的系統狀況選擇適合的派工法則並且建立一個動態的派工法則知識庫則會使得彈性製造系統的產出提高。為了提供彈性製造系統一個可以選取最佳系統資訊並且有高歸納力的分類器，本篇論文提出了一個自我組織的派工模型。為了達到上述的目標，本論文所提出的派工系統模型將會包含了模糊理論、基因演算法、還有類神經網路。除此之外，本研究更利用模擬軟體建立了一個彈性製造系統，並且在模擬實驗證明本論文所提出的動態派工模型相較於單一派工法則可以得到較優產出。With the selection of the real-time salient information of machines and parts and then a rule’s dispatching mechanism is built for the scheduling task, the dynamic scheduling rules would outperform static ones in a flexible manufacturing system (FMS). Scheduling plays an important role in the production control in a foundry fab, which can be seen as a huge FMS. For a dynamic scheduling system, two critical issues dominate the performance of a scheduled FMS, one is the selection of system attributes and the other is the design of the dispatching mechanism, namely, the classifier design. This thesis proposes a self-organizing scheduling model (SOSM) aiming to provide a FMS with the optimal attribute selection and high generalization, high-accuracy classifier. The proposed scheduling model combines several intelligent methods to achieve these goals, including fuzzy set theory, genetic algorithms, and neural networks. A typical FMS model is conducted in the experiments for the demonstration. Experimental results show that the SOSM outperforms the static dispatching rules under three different performance criteria.中文摘要 i
Abstract ii
Contents . iv
List of Tables v
List of Figures vi
Chapter 1 Introduction 1
1.1 Motivation 1
1.2 Literature Survey 2
1.2.1 FMS 2
1.2.2 Dynamic Scheduling Problems of a FMS 3
1.3 Contributions 4
1.4 Thesis Organization 5
Chapter 2 Background Knowledge 6
2.1 Overview of Machine Learning and Intelligent Scheduling 6
2.1.1 Applying Machine Learning to Intelligent Scheduling 6
2.2 Attribute selection approaches 8
2.3 Support Vector Machine 10
2.3.1 Linearly separable SVM 10
2.3.2 Linearly nonseparable SVM 11
2.3.3 Nonlinear SVM 14
2.3.4 Multi-class Support Vector Machines 16
Chapter 3 Selecting Optimal System Attributes with GA-based FFEI 20
3.1 Introduction 20
3.1.1 Supervised Feature Mining (SFM) 21
3.1.2 Relative Importance and Weighting Factors 21
3.2 Fuzzy entropy based feature evaluation index 23
3.2.1 Minimizing the FFEI via GA 28
3.3 Experiment result of feature selection 29
3.3.1 Validity demonstration with classification accuracy 32
Chapter 4 Scheduler Design Using Support Vector Machines 34
4.1 Introduction 35
4.1.1 Scheduler training part 36
4.2 Construct the Scheduling Knowledge Base 38
4.3 Adjusting Dispatching Rule with Voting Strategy 41
4.3.1 One-against-one Multi-classification SVM 41
4.3.2 Voting Strategy 41
4.4 Preparing the Training Data 43
4.5 On-line Scheduling Part 43
Chapter 5 Experiments 45
5.1 Layout and Assumptions of FMS 45
5.1.1 Training Set Preparation 51
5.1.2 Testing Set Preparation 53
5.1.3 Data Scaling 53
5.2 Experimental Results 53
Chapter 6 Conclusion 67
6.1 Future work 6
Thansamay, Pai Thiaw, Tham Bun:ransnational Modernity of Thai-Isan Migrant Workers
No full text 台灣的跨國移工研究，多著重跨國移工於台灣社會內部所遇限制，及遭逢勞動控制所形成的反抗，忽略了跨國移工處於跨國場域中，因而缺乏跨國主義取向的研究。本研究從跨國主義觀點出發，討論位於兩地間的跨國連結。以「現代性」為主題，本研究認為，跨國連結不只是鉅觀經濟結構限制，與微觀對於城市的想像，更須從地方社群的社會關係，及其文化意涵出發，始能理解跨國連結現象，在這些不同層次的再連結。 以泰國東北Isan勞工為研究對象，本研究首先從政治經濟學出發，探討跨國移動形成的客觀因素。勞動市場分割，與跨國移工生產所得，與再生產消費的分離，促成跨國移動的背景。而泰國東北地區，與全球各地的政經變化，使Isan移工們於不同時代，前往相異地區工作。 客觀的政治經濟結構，對於行動者的意識假設過於簡單。在泰國東北，學者指出泰國內部的政經變化，造成Isan人們對於「城市現代性」的嚮往，因而前往都市或繁華他國工作。但本研究認為，城鄉區隔遠比此單線嚮往複雜，Isan移工們往往處於兩地矛盾間，既期待城市生活，卻又對東北家鄉有所依戀。 處於現代兩地矛盾間的Isan移工，須從地方的社會關係及其文化意涵了解。泰國東北Isan女性，符合東北女性對於家庭的經濟角色，而外出工作。另一方面，她也運用了「趕得上時代」(ทันสมัย thansamay)的論述，以此新建立的社會關係，形成對於家鄉性別控制的反抗。 面臨女性遠行威脅，與對其性解放的擔憂，男性跨國移工的出現，可視為此一脈絡下的回應。在返鄉後，男性以泰國東北文化中，強調男性正面移動性的「遠行」(ไปเที่ยว pai thiaw)論述，以及交織著個人威望及社會關係的「作功德」(ทำบุญ tham bun)實踐，重申其在泰國東北社會中的地位。 Studies of transnational migrant workers in Taiwan put more emphasis on their limitations in Taiwan, and their resistances facing labor controls. Those studies often ignore transnational workers are locating in a transnational field. Therefore, there are few “transnationalism” studies. From transnationalism, this study discusses connections of transnational field. Take “modernity” as the main issue, this study argues, the transnational connections are not only formed by macro political economy or micro imagination of urban life. We should pay more attention to local social relations and their cultural meanings, and reconnect those connections of different levels. First, from aspects of political economy, this study examines the objective factors forming movement of Thai-Isan workers. The division of labor market and the separation of reproduction from production constitute the background of transnational movement. Besides, Thai and global politico-economic change push Thai-Isan workers to work in different regions. Studies of political economy often simplify actors’ motivations. In Thai-Isan, scholars indicate, the political economic changes cause the desires of ‘urban modernity’, and push Thai-Isan workers to work in the city or abroad. However, this study argues, the rural-urban division is more complex than one-way desire. Thai-Isan workers are often situated in the dilemma of urban life and their northeastern hometown. The dilemma of Thai-Isan migrant workers should be understood with local social relations and cultural meanings. Thai-Isan women work outside fitting economic roles of women in Thai-Isan community. Hoever, they use discourses of “up-to-date” (ทันสมัย thansamay) to form new social relations outside their hometown, and to resist gender control with their home community. Facing threat of women’s mobility and their sexual freedom, the emergence of transnational Thai-Isan male workers could be seen as a reaction to this context. After returning home, men use the discourses of “going playing” (ไปเที่ยว pai thiaw) to emphasize their positive experiences of mobility. Besides, they spend money they earned outside to “make merit” (ทำบุญ tham bun). Those discourses and practices, which are crucial in Thai-Isan socio-cultural life, are used by transnational Thai-Isan male workers to reclaim their status within social relations in Thai-Isan community.口試委員會審定書 ……………………ii謝 ……………………………………iii文摘要 ………………………………vi文摘要 ………………………………vii錄 ……………………………………viii表目錄 ………………………………x一章 導言……………………………01二章 政治經濟………………………11第一節 「工作」意涵………………11第二節 地方經濟……………………12第三節 情勢變化……………………18三章 城鎮之間………………………23第一節 論「現代性」………………24第二節 嚮往城市……………………26第三節 依戀家鄉……………………29四章 「現代」東北…………………33第一節 東北男女……………………36第二節 社會緊張……………………39第三節 「現代」女性………………43第四節 男性回應……………………45五章 「現代」男性…………………47第一節 行動意義……………………47第二節 遠行經驗……………………54第三節 功德儀式……………………60六章 討論與結論……………………70跋：誰的跨國現代性？ ……………78用書目 ………………………………85錄 ……………………………………99附錄一 泰文─中文對照 ……………99附錄二 泰國曼谷各種職業收入……101附錄三 老鼠唧與老鼠啁……………103附錄四 大學之星……………………10
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