31 research outputs found

    Space-decomposition multiplier method for constrained minimization problems

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    AbstractIn this paper, a new multiplier method that decomposes variable space into decomposed spaces is introduced. This method allows constrained minimization problems to be decomposed into subproblems. A potential constraint strategy that uses only part of the constraint set in the decomposed-space subproblems is also presented to increase the efficiency of this new space-decomposition multiplier method. Three examples are given to demonstrate this method and the potential constraint strategy

    Identification of the genetic determinants of Salmonella enterica serotype Typhimurium that may regulate the expression of the type 1 fimbriae in response to solid agar and static broth culture conditions

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    <p>Abstract</p> <p>Background</p> <p>Type 1 fimbriae are the most commonly found fimbrial appendages on the outer membrane of <it>Salmonella enterica </it>serotype Typhimurium. Previous investigations indicate that static broth culture favours <it>S</it>. Typhimurium to produce type 1 fimbriae, while non-fimbriate bacteria are obtained by growth on solid agar media. The phenotypic expression of type 1 fimbriae in <it>S</it>. Typhimurium is the result of the interaction and cooperation of several genes in the <it>fim </it>gene cluster. Other gene products that may also participate in the regulation of type 1 fimbrial expression remain uncharacterized.</p> <p>Results</p> <p>In the present study, transposon insertion mutagenesis was performed on <it>S</it>. Typhimurium to generate a library to screen for those mutants that would exhibit different type 1 fimbrial phenotypes than the parental strain. Eight-two mutants were obtained from 7,239 clones screened using the yeast agglutination test. Forty-four mutants produced type 1 fimbriae on both solid agar and static broth media, while none of the other 38 mutants formed type 1 fimbriae in either culture condition. The flanking sequences of the transposons from 54 mutants were cloned and sequenced. These mutants can be classified according to the functions or putative functions of the open reading frames disrupted by the transposon. Our current results indicate that the genetic determinants such as those involved in the fimbrial biogenesis and regulation, global regulators, transporter proteins, prophage-derived proteins, and enzymes of different functions, to name a few, may play a role in the regulation of type 1 fimbrial expression in response to solid agar and static broth culture conditions. A complementation test revealed that transforming a recombinant plasmid possessing the coding sequence of a NAD(P)H-flavin reductase gene <it>ubiB </it>restored an <it>ubiB </it>mutant to exhibit the type 1 fimbrial phenotype as its parental strain.</p> <p>Conclusion</p> <p>Genetic determinants other than the <it>fim </it>genes may involve in the regulation of type 1 fimbrial expression in <it>S</it>. Typhimurium. How each gene product may influence type 1 fimbrial expression is an interesting research topic which warrants further investigation.</p

    Lovastatin Modulates Glycogen Synthase Kinase-3β Pathway and Inhibits Mossy Fiber Sprouting after Pilocarpine-Induced Status Epilepticus

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    This study was undertaken to assay the effect of lovastatin on the glycogen synthase kinase-3 beta (GSK-3β) and collapsin responsive mediator protein-2 (CRMP-2) signaling pathway and mossy fiber sprouting (MFS) in epileptic rats. MFS in the dentate gyrus (DG) is an important feature of temporal lobe epilepsy (TLE) and is highly related to the severity and the frequency of spontaneous recurrent seizures. However, the molecular mechanism of MFS is mostly unknown. GSK-3β and CRMP-2 are the genes responsible for axonal growth and neuronal polarity in the hippocampus, therefore this pathway is a potential target to investigate MFS. Pilocarpine-induced status epilepticus animal model was taken as our researching material. Western blot, histological and electrophysiological techniques were used as the studying tools. The results showed that the expression level of GSK-3β and CRMP-2 were elevated after seizure induction, and the administration of lovastatin reversed this effect and significantly reduced the extent of MFS in both DG and CA3 region in the hippocampus. The alteration of expression level of GSK-3β and CRMP-2 after seizure induction proposes that GSK-3β and CRMP-2 are crucial for MFS and epiletogenesis. The fact that lovastatin reversed the expression level of GSK-3β and CRMP-2 indicated that GSK-3β and CRMP-2 are possible to be a novel mechanism of lovatstain to suppress MFS and revealed a new therapeutic target and researching direction for studying the mechanism of MFS and epileptogenesis

    Study of Protein Cross-Links in Copulatory Plughe Major TG4 Cross-linking Sites of SVS I in mouse Seminal Vesicle

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    小鼠貯精囊分泌液以還原性 SDS-PAGE 電泳分析,可以觀察到七個明顯的蛋白,按照分子量大小命名為 SVS I-VII。而實際上在貯精囊液中幾乎沒有 SVS I、SVS II、和SVS III以單體形式存在,它們會以雙硫鍵連結成分子量非常大的不同蛋白質複合體 (high molecular weight complexes, HMWCs)。經由免疫組織染色法證明 SVS I、SVS II、SVS III蛋白是組成交配栓的蛋白成分。將交配栓以還原性SDS-PAGE sample buffer 粹取,幾乎沒有 SVS I 和 SVS II 被溶解出來,只有少量 SVS III 會被溶解出來,顯示在交配栓中 SVS I-III 之間除了雙硫鍵之外還有其他共價鍵參與其中。進一步純化凝固腺中轉麩胺醯酶 (TG4),並且與組織內的轉麩胺醯酶 (TG2) ,比較催化貯精囊分泌蛋白的酵素活性。我們發現 TG4 對於 HMWCs 具有較高的活性。然而 TG2 對於還原後貯精囊液中自由的 SVS I-III 蛋白具有較佳的活性。這種高效率的 TG4 催化在,精液凝固而形成交配栓是一重要的生化反應,其生殖的意義會加以討論。VS I 蛋白質序列共有 820 個胺基酸,其中 43 個為麩胺酸 43 個為離胺酸。,我們根據其序列製備 7 段重組蛋白,分成 1-78 胺基酸序列為 F1 片段、79-259 胺基酸序列為 F2 片段、260-405 胺基酸序列為 F3 片段、406-500 胺基酸序列為 F4 片段、501-650 胺基酸序列為 F5 片段、651-715 胺基酸序列為 F6 片段以及 716-796 胺基酸序列為 F7 片段。比較 F1~F7 片段被 TG4 催化後所接上 BPNH2 或 A25 peptide的數量。F2 片段對於 BPNH2 作用的活性遠大於其他片段,而 A25 peptide 對於各片段的反應活性相對較低。質譜分析經 TG4 催化,而連結上 BPNH2 的 F2 片段,鑑定出 Q232 和 Q254 是TG4 作用標的。將 F2 片段的 Q232 和 Q254 突變為三種 F2 片段 (Q232G、Q254G 和Q232G/Q254G),BPNH2 連接上去的數量相對原始 F2 片段減少,確認 Q232 和 Q254 為 TG4 催化的主要目標。Resolution of mouse seminal vesicle secretion (SVS) by reducing SDS-PAGE can clearly identified seven major monomer proteins tentatively designated as SVS I-VII according to the decreasing order of their molecular masses. However, no monomer forms of SVS I-III are present in SVS. Instead, they appear in various high molecular weight complexes (HMWCs) formed by inter-polypeptide disulfide bridges among SVS I, SVS II and SVS III. The HMWCs become the predominant protein in SVS. We were able to immunodetect SVS I-III in cross sections of mouse copulatory plug. Heating the clotted lump in SDS-PAGE sample buffer in the presence of DTT released none of SVS I or SVS II but only a trace of SVS III into the solution, indicative of covalent cross-links in addition to disulfide bonds among the SVS I-III proteins in the plug. Further, we purified type 4 transglutaminase (TG4) from the mouse coagulating gland to homogeneity, and compared it to type 2 transglutaminase (TG2) from guinea pig liver in terms of enzymatic ability to cross-link proteins in the SVS. We found that TG4 showed much greater activity than TG2 in catalyzing the cross-links between HMWC proteins. On the contrary, it was less active than TG2 in cross-linking any of the free SVS I-III proteins released from the reduced HMWC. The reproductive significance of more efficient TG4 catalysis is discussed.ased on the SVSI-deduced protein sequence, SVS I contains 820 amino acid residues in which there are 43 glutamine and 43 lysine residues. We produced 7 recombinant polypeptide fragments including residues 1-78/F1, residues 79-259/F2, residues 260-405/F3, residues 406-500/F4, residues 501-650/F5, residues 651-715/F6, and residues 716-796/F7, and measured the covalent incorporation of 5-(biotinamido)pentylamine (BPNH2) or biotin-TVQQEL (A25 peptide) to each of F1-to-F7 by TG4. F2 was much more active than the other fragments during the BPNH2-glutamine incorporation, Relatively, a low extent of A25 cross-linking to any one of the seven polypeptide fragments was observed. The MS analysis of BPNH2-F2 conjugate identified Q232 and Q254 as the two major TG4 cross-linking sites. This was substantiated by the result that much less BPNH2 was cross-linked to any one of three F2 mutants, including Q232G and Q254G obtained from single-site mutation, and Q232G/Q254G from double-site mutation.縮寫表 III要 IV bstract V 一章 緒論 1.1 生殖學概論 1.2 哺乳類雄性生殖系統 1.3 附屬性腺的研究 2.3.1 附屬性腺的差異 2.3.2 附屬性腺的生理功能 3.3.3 附屬性腺的病理研究 4.4 前列腺的發育與功能 4.5 貯精囊發育與功能 4.6 交配栓 6.6.1 交配栓的形成 6.6.2 交配栓的功能 6.7 轉麩胺醯胺酶的功能 7.7.1 轉麩胺醯胺酶的分類 7.7.2 轉麩胺醯胺酶催化的轉化醯基反應 8.7.3 轉麩胺醯胺酶的活性反應區和結構 8.7.4轉麩胺醯胺酶的基質選擇性 9.8 研究背景 10.8.1小白鼠貯精囊 SVS I 之研究 10.8.2交配栓內貯精囊液蛋白的共價鍵結 11二章 材料與方法 12.1實驗材料 12.1.1 化學試劑與酵素 12.1.2 實驗動物 12.2 SVS I-III 抗體製備 12.2.1 重組質體的製備 12.2.2 重組蛋白的純化 13.3 免疫組織染色 13.4 對角線電泳分析 14.5 小鼠 TG4 純化 14.6 Solid-Phase Microtiter Assay 15.7 蛋白質交聯實驗 15.8 SVS I F1-F7 片段的製備 15.8.1 重組質體的製備 15.8.2 重組蛋白的純化 16.9 製備接上 BPNH2 的 SVS I F2 片段 17三章 實驗結果 18.1交配栓內貯精囊液蛋白的共價鍵結 18.1.1小鼠 SVS I、SVS II、SVS III 抗體製備 18.1.2 SVS I-III是交配栓的組成成分8.1.3 貯精囊分泌蛋白 SVS I-III 會形成高分子聚合物 18.1.4交配栓內 SVS I-III 之間共價鍵結 19.1.5 生殖系統特有的轉麩胺醯酶 19.1.6 純化及鑑定小鼠凝固腺分泌蛋白TG4 20.1.7 TG4酵素活性測定 20.1.8 在貯精囊分泌蛋白中SVS I-III是TG4的基質 21.1.9 HMWCs 分子間雙硫鍵對於TG作用的影響 21.2 鑑定貯精囊蛋白SVS I被凝固腺液之轉麩胺醯酶催化的活性部位 22.2.1 製備SVS I F1~F7片段重組蛋白 22.2.2 鑑定 SVS I 被催化的活性部位 22四章 討論 58.1 SVS I-III 參與交配栓的形成 58.2 TG4 催化 HMWCs 內的蛋白鍵結 58.3 小鼠 SVS I 與 TG 的作用 59eferences 6

    A comparative analysis of data mining techniques for prediction of postprandial blood glucose: A cohort study

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    The use of advanced predictive techniques and reasoning models has greatly assisted clinicians in improving the diagnosis, prognosis, and treatment of diabetes. Although numerous studies have focused on the relationship between abnormal blood glucose levels and diabetes, few have focused on the risk forecasting of postprandial blood glucose levels in patients with diabetes. This work aimed to develop a model for the prediction of postprandial blood glucose levels to screen for undiagnosed diabetes cases in a cohort study. The performance of the proposed model was then compared with those of five other datamining techniques: random forest (RF), support vector machine (SVM), C5.0, multilayer perceptron (MLP), and logistic regression (LR). The data of 1,438 patients who were admitted to Landseed Hospital, Northern Taiwan, over the period of 2006 and 2013 were collected and used to evaluate the performances of the data-mining techniques. Compared with the 4.5, SVM, MLP, and LR models, the RF model had the best prediction capability for postprandial blood glucose levels in terms of the overall correct classification rate. The results of this study underscore the importance of identifying the preclinical symptoms of abnormal blood glucose levels. The proposed model provides precise reasoning and prediction and can be used to help physicians improve the diagnosis, prognosis, and treatment of patients with diabetes

    Risk Assessment of Coastal Flooding under Different Inundation Situations in Southwest of Taiwan (Tainan City)

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    The Pacific island countries are particularly vulnerable to the effects of global warming including more frequent and intense natural disasters. Seawater inundation, one of the most serious disasters, could damage human property and life. Regional sea level rise, highest astronomic tide, vertical land motions, and extreme sea level could result in episodic, recurrent, or permanent coastal inundation. Therefore, assessing potential flooding areas is a critical task for coastal management plans. In this study, a simulation of the static flooding situation in the southwest coast of Taiwan (Tainan city) at the end of this century was conducted by using a combination of the Taiwan Digital Elevation Model (DEM), regional sea level changes reconstructed by tide gauge and altimetry data, vertical land deformation derived from leveling and GPS data, and ocean tide models. In addition, the extreme sea level situation, which typically results from high water on a spring tide and a storm surge, was also evaluated by the joint probability method using tide gauge records. To analyze the possible static flood risk and avoid overestimation of inundation areas, a region-based image segmentation method was employed in the estimated future topographic data to generate the flood risk map. In addition, an extreme sea level situation, which typically results from high water on a spring tide and a storm surge, was also evaluated by the joint probability method using tide gauge records. Results showed that the range of inundation depth around the Tainan area is 0–8 m with a mean value of 4 m. In addition, most of the inundation areas are agricultural land use (60% of total inundation area of Tainan), and two important international wetlands, 88.5% of Zengwun Estuary Wetlands and 99.5% of Sihcao Wetlands (the important Black-faced Spoonbills Refuge) will disappear under the combined situation. The risk assessment of flooding areas is potentially useful for coastal ocean and land management to develop appropriate adaptation policies for preventing disasters resulting from global climate change

    Drug administration regimen.

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    <p>All the drugs were injected subcutaneously after SE induction.</p

    The experimental design and Timm’s score recorded from different time points.

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    <p>(A) The experimental design of TLE animal model and drug administration. (B) Timm’s score recorded from control and 1 to 3 months after SE induction. 1 month after SE induction, the Timm’s score was significantly increased compared with control group and show an increasing tendency with time. (*compared with control group).</p
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