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Identification of active agents inducing ossification and novel biomarkers associated with osteoporosis

為了解決Rp Wine rating的問題，作者Zuha Imtiyaz 這樣論述：

TABLE OF CONTENTSABSTRACT ixLIST OF ABBREVIATIONS xi1. INTRODUCTION 11.1 Bone and bone remodeling 11.2 Osteoporosis 61.2.1 Disease epidemiology 71.2.2 Current drugs used for osteoporosis and their mode of action 91.3 Biomarkers of bone formation 121.3.1 SNPs as biomarkers 161.4 Relation of

FTCDNL1 gene with osteoporosis 191.5 Herbs with medicinal value for bone formation 20RATIONALE 25AIM 262. MATERIALS AND METHODS 272.1 Reagents and chemicals used 272.2 Extraction and isolation 292.2.1 Turpinia formosana Nakai. 292.2.2 Euonymus spraguei Hayata. 292.3 Cell culture 302.3.1 H

uman osteoblast (HOb) cells 302.3.2 Human osteosarcoma (MG-63) cells 302.3.3 Cell culture of RAW 264.7 cells and differentiated osteoclasts 312.4 MTT assay for cell viability assay 312.5 ALP activity assay 312.6 Mineralization assay 322.7 RNA isolation and reverse transcription 322.8 Real-tim

e quantitative PCR analysis 332.9 Estrogen receptor expression assay 352.10 Plasmid DNA purification 352.11 DNA transfection 362.12 siRNA transfection 362.13 Western blot 372.14 Immunofluorescence 372.15 Statistical Analysis 383. RESULTS 393.1 Elucidating osteogenic potential of compounds i

solated from Turpinia formosana Nakai 393.1.1 Isolated compounds from Turpinia formosana 393.1.2. Cytotoxicity of compounds isolated from T. formosana in HOb cells 413.1.3 Effect of isolated compounds on ALP activity in HOb cells 433.1.4 Effect of isolated compounds on mineralization in HOb cell

s 453.1.5 Effect of isolated compounds on estrogen receptor expression in HOb cells 473.1.6 Effect of isolated compounds on the genetic markers of bone formation in HOb cells 493.2. Isolated compounds from Euonymus spraguei mediate of osteogenesis through multiple pathways 523.2.1 Effect of vari

ous extractants for E. spraguei on cell viability, ALP activity and mineralization in HOb cells 523.2.2 Compounds isolated from ES 563.2.3 Effect of syringin (8) and (-)-epicatechin (9) on the viability of HOb cells 573.2.4 Effect of syrngin (8) and (-)-epicatechin (9) on ALP activity in HOb cell

s 583.2.5 Effect of syrngin (8) and (-)-epicatechin (9) on mineralization in HOb cells 603.2.6 Effect of syrngin (8) and (-)-epicatechin (9) on Estrogen receptor (ER) expression 623.2.7 Effect of syringin (8) and (-)-epicatechin (9) on the mRNA expression levels of bone remodelling-related genes

633.2.8 Effect of syringin (8) and (-)-epicatechin (9) on BMP-2 pathway 663.2.9 Effect of syringin (8) and (-)-epicatechin (9) on RANKL/OPG 693.2.10 Effect of syringin (8) and (-)-epicatechin (9) on autophagy 713.2.11 Effect of syringin (8) and (-)-epicatechin (9) on OPN expression 733.2.12 Eff

ect of syringin (8) and (-)-epicatechin (9) on the viability of RANKL-induced osteoclasts 753.3. Elucidating the association between FTCDNL1 on bone formation and its role in osteoporosis onset 773.3.1 Transfection of FTCDNL1 into MG-63 cells 773.3.2 Effect of FCTDNL1 overexpression on bone forma

tion-related genes 793.3.3 Effect of FTCDNL1 knockdown on cell viability of MG63 cells 833.3.4 Effect of FTCDNL1 knockdown on ALP activity in MG63 cells 844. DISCUSSION 854.1. Elucidating osteogenic potential of compounds isolated from Turpinia formosana 874.2 Isolated compounds from Euonymus

spraguei mediate of osteogenesis through multiple pathways 904.3. Elucidating the association between FTCDNL1 on bone formation and its role in osteoporosis onset 945. CONCLUSION 96LIST OF PUBLICATIONS 97BIBLIOGRAPHY 98 LIST OF FIGURESFig 1. Types of cells found within the bone tissue 1Fig 2.

Interplay between osteoblasts and osteoclasts 3Fig 3. Involvement of various cells at different stages of bone remodelling 4Fig 4. Static and dynamic changes in bone 7Fig 5. Growth rate of osteoporosis, estimated number of osteoporotic fracture worldwide by 2050 9Fig 6. Mechanism of action of th

e current drugs for osteoporosis. 11Fig 7. Role of various biomarkers at various stages of bone formation 14Fig 8. Signalling cascade of BMP-2 pathway. 16Fig 9. Display of the importance of a SNP and its significance in the phenotype 17Fig 10. Pictorial representation of aim of the study. 26Fig

11. Structures of the isolated compounds elucidated using various spectroscopic techniques. 40Fig 12. Cytotoxic effect of compounds isolated from T.formosna. 41Fig 13. Induction of ALP activity by 1, 2, 3 and 6. 43Fig 14. Mineral deposition by 1, 2, 3 and 6. 46Fig 15. Expression of ERs affected

by 1, 2, 3, and 6. 48Fig 16. 1 elevates mRNA expression of bone formation-related genes. 50Fig 17. Pictorial summary of underlying mechanism for osteogenesis by 1. 51Fig 18. Solvent test for extraction of E. spraguei (ES). 54Fig 19. Structure of compounds isolated from ES. 56Fig 20. Cell viabi

lity of HOb cells on 8 and 9 treatments. 57Fig 21. Induction of ALP activity by 8 and 9. 58Fig 22. Mineral deposition by 8 and 9.. 61Fig 23. Influence of 8 and 9 on ER expression. 62Fig 24. Expression of bone formation-related genes after 8 and 9 treatments.. 65Fig 25. 8 and 9 mediates their ef

fect via BMP-2 signalling pathway. 67Fig 26. Attenuation of RANKL/OPG ratio by 8 and 9. 69Fig 27. Modulation of autophagy by 8 and 9. 72Fig 28. Induction of OPN by 8 and 9. 74Fig 29. Cytotoxic effect of 8 and 9 on osteoclasts. 75Fig 30. Pictorial summary of osteogenesis induced by 8 and 9. 76F

ig 31. Transient transfection of FCTDNL1 in MG63 cells. 78Fig 32. mRNA expression levels of bone formation related biomarkers after FTCDNL1 overexpression. 81Fig 33. FTCDNL1 knockdown does not affect viability of MG63 cells. 83Fig 34. ALP activity of MG63 cells after FTCDNL1 knockdown. 84Fig 35.

Mechanism of 8 and 9 in RANK-RANKL interaction 92Fig 36. Role of autophagy in bone formation. 93Fig 37. Pictorial representation of osteogenesis with FTCDNL1 and future prospects. 95LIST OF TABLESTable 1. Primer and probe combination used for real-time PCR 34Table 2. Primer sequences used for r

eal-time PCR 35Table 3. Sequences of siRNA used for FTCDNL1 knockdown 37Table 4. Phytochemical as therapeutic agents and their plant source 85

不同加工方法與配方對傳統印尼糯米糕感官特性之影響

為了解決Rp Wine rating的問題，作者鄧金蓮 這樣論述：

摘要印尼傳統的糯米糕 (Klepon) 在印尼是一個最有名的傳統點心，來自於爪哇，是一個內餡包黑糖而外面用椰絲包覆，通常是烹煮過後食用的甜點。他是印尼的一種美食遺產，推展到其他國家將是一項重要的工作。在全球市場擴張的時代，跨文化研究與科學及快速的感官品評評估技術得到了更多的注意，因為研究發現文化因素對食品的接受性和喜好性有顯著性的影響。有別於傳統的描述分析技術，選擇適合項目法(CATA)是一種以消費者品評員進行食品描述分析的快速方法，已被大量使用在評估產品開發和配方的確定及為行銷目的定義消費者有感的關鍵感官特性。本研究的目的是利用9分法和選擇適合分析法評估3個因子所製備的12種糯米糕，包含不

同種類的糯米粉(印尼糯米粉，台灣長糯米粉和台灣圓糯米粉), 糯米粉百分比(100％ 和 90％添加了10％湯種米麵團)及不同的烹飪方法(煮和蒸)對產品的感官接受性和物理特性的影響，同時了解印尼和台灣消費者對於感官特性感覺上的差異。本研究招募了120位印尼當地消費者(60位評估煮及60位評估蒸的樣品)使用傳統品評單，和136位台灣消費者(71位評估煮及65位評估蒸的樣品)於品評小室使用電腦化的品評單，評估消費者的喜歡程度、24種感官屬性、5種感官感覺與購買意圖。樣品提供順序使用拉丁威廉方塊設計。質地分析和顏色範圍使用質地分析儀和色差儀進行分析。研究結果顯示糯米粉的種類和烹飪方法與糯米粉百分比的交

互作用影響了印尼消費者對產品的消費者接受性；台灣消費者對產品的消費者接受性則受到糯米粉的種類、糯米粉百分比與烹飪方法與糯米粉百分比交互作用的影響。台灣消費者的9分法測試的平均值比印尼消費者低，使用Tukey’s HSD，13個印尼糯米糕的消費者接受程度沒有顯著差異(p>0.05)。選擇適合項目法的結果顯示印尼消費者能夠顯著的區分出樣品的感官特性(p