Wan Tai Fung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 102-111). / Abstracts in English and Chinese. / Thesis committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Chinese Abstract --- p.v / Acknowledgements --- p.vii / General Abbreviations --- p.ix / Abbreviations of Chemicals --- p.xi / Table of Contents --- p.xii / List of Figures --- p.xvi / List of Tables --- p.xvi / Chapter 1 --- Literature Review --- p.1 / Chapter 1.1 --- Soybeans --- p.1 / Chapter 1.1.1 --- Nutrient composition of soybean --- p.1 / Chapter 1.1.2 --- Nitrogen fixation and assimilation in soybean --- p.3 / Chapter 1.1.3 --- The role in nitrogen allocation and controlling the nitrogen sink-source relationship of asparagine --- p.3 / Chapter 1.1.4 --- Characterization of asparagine synthetase --- p.8 / Chapter 1.1.4.1 --- Biochemistry and molecular background of plant asparagine synthetase --- p.8 / Chapter 1.1.4.2 --- Asparagine synthetase in Arabadopsis thaliana --- p.9 / Chapter 1.1.4.3 --- "Asparagine synthesis in soybean, Glycine max" --- p.10 / Chapter 1.1.4.4 --- "Asparagine synthetase in rice, Oryza sativa" --- p.11 / Chapter 1.2 --- Seed protein quality and quantity improvement --- p.13 / Chapter 1.2.1 --- Nutrition composition of rice --- p.13 / Chapter 1.2.2 --- Molecular approaches for improving seed storage protein quality --- p.14 / Chapter 1.2.2.1 --- Protein sequence modification --- p.14 / Chapter 1.2.2.2 --- Synthetic genes --- p.16 / Chapter 1.2.2.3 --- Overexpression of homologous genes --- p.17 / Chapter 1.2.2.4 --- Transfer and expression of heterologous genes --- p.18 / Chapter 1.2.2.5 --- "Manipulation of pathway synthesizing essential amino acids, aspartate family amino acid" --- p.19 / Chapter 1.2.3 --- Research in improving rice seed protein quality and quantity --- p.22 / Chapter 1.3 --- Hypothesis and objective of this study --- p.23 / Chapter 2 --- Materials and Methods --- p.25 / Chapter 2.1 --- Materials --- p.25 / Chapter 2.1.1 --- Plant materials --- p.25 / Chapter 2.1.2 --- Bacterial strains and vectors --- p.26 / Chapter 2.1.3 --- Growth conditions for soybean --- p.26 / Chapter 2.1.4 --- Chemicals and reagents --- p.26 / Chapter 2.1.5 --- "Buffer, solution and gel" --- p.26 / Chapter 2.1.6 --- Commercial kits --- p.27 / Chapter 2.1.7 --- Equipments and facilities used --- p.27 / Chapter 2.1.8 --- Primers --- p.27 / Chapter 2.2 --- Methods --- p.28 / Chapter 2.2.1 --- Growth condition for plant materials --- p.28 / Chapter 2.2.1.1 --- General conditions for planting soybean --- p.28 / Chapter 2.2.1.2 --- Soybean seedlings for gene expression profile analysis --- p.28 / Chapter 2.2.1.3 --- Mature soybean for gene expression profile analysis --- p.29 / Chapter 2.2.1.4 --- Mature soybean for cloning of AS I and AS2 full length cDNA --- p.30 / Chapter 2.2.1.5 --- Mature soybean seed for amino acid profile analysis --- p.30 / Chapter 2.2.1.6 --- General conditions for planting transgenic rice in CUHK --- p.30 / Chapter 2.2.1.7 --- Transgenic rice seedling for PCR screening --- p.31 / Chapter 2.2.1.8 --- Transgenic rice for functional test and seed for biochemical analysis --- p.31 / Chapter 2.2.2 --- Molecular techniques --- p.32 / Chapter 2.2.2.1 --- Total RNA extraction --- p.32 / Chapter 2.2.2.2 --- Denaturing gel electrophoresis for RNA --- p.33 / Chapter 2.2.2.3 --- Northern blot analysis --- p.33 / Chapter 2.2.2.3.1 --- Chemiluminescent detection --- p.33 / Chapter 2.2.2.3.2 --- Film development --- p.34 / Chapter 2.2.2.4 --- Preparation of single-stranded DIG-labeled PCR probes --- p.34 / Chapter 2.2.2.4.1 --- Primer design for the PCR probes of --- p.34 / Chapter 2.2.2.4.2 --- Amplification of AS1 and AS2 internal PCR fragments --- p.34 / Chapter 2.2.2.4.3 --- Quantitation of purified AS1 and AS2 PCR fragments --- p.35 / Chapter 2.2.2.4.4 --- Biased PCR to make single-stranded DNA probes --- p.35 / Chapter 2.2.2.4.5 --- Probe quantitation --- p.36 / Chapter 2.2.2.5 --- Probe specificity test --- p.37 / Chapter 2.2.2.6 --- Cloning of full length cDNA --- p.37 / Chapter 2.2.2.6.1 --- First strand cDNA synthesis from RNA of high protein content soybean leaf --- p.37 / Chapter 2.2.2.6.2 --- PCR for amplification of AS1 and AS2 full length cDNA --- p.38 / Chapter 2.2.2.6.3 --- Preparation of pBluescript II KS(+) T-vector for cloning --- p.38 / Chapter 2.2.2.6.4 --- Ligation of DNA inserts into pBluescript II KS(+) T-vector --- p.39 / Chapter 2.2.2.6.5 --- Preparation of E. coli DH5α CaCl2-mediaed competent cells --- p.39 / Chapter 2.2.2.6.6 --- Transformation of E. coli DH5α competent cell --- p.40 / Chapter 2.2.2.7 --- Screening of recombinant plasmids --- p.40 / Chapter 2.2.2.7.1 --- Isolation of recombinant plasimid DNA from bacterial cells --- p.41 / Chapter 2.2.2.7.2 --- PCR screening on recombinant plasmids --- p.41 / Chapter 2.2.2.7.3 --- DNA gel electrophoresis --- p.41 / Chapter 2.2.2.8 --- Sequencing and homology search --- p.42 / Chapter 2.2.2.9 --- Functional test using transgenic plant --- p.43 / Chapter 2.2.2.9.1 --- Preparation of chimeric gene constructs and recombinant plasmids --- p.43 / Chapter 2.2.2.9.2 --- Agrobacterium mediated transformation into rice calli to regenerate transgenic AS1/ AS2 rice --- p.44 / Chapter 2.2.2.10 --- PCR Screenig of homozygous and heterozygous transgenic plants --- p.44 / Chapter 2.2.2.10.1 --- Isolation of genomic DNA from transgenic plants --- p.45 / Chapter 2.2.2.10.2 --- PCR screening using genomic DNA --- p.46 / Chapter 2.2.2.11 --- Quantitative PCR analysis on transgenic plants --- p.48 / Chapter 2.2.3 --- Biochemical Analysis --- p.49 / Chapter 2.2.3.1 --- Quantitative amino acid analysis in mature soybean seeds --- p.49 / Chapter 2.2.3.2 --- Quantitative amino acid analysis in mature transgenic rice grain --- p.49 / Chapter 3 --- Results --- p.50 / Chapter 3.1 --- Amino acid analysis on mature soybean seeds --- p.50 / Chapter 3.2 --- Expression pattern analysis of AS genes by Northern Blot analysis --- p.54 / Chapter 3.2.1 --- Making of single strand digoxigenin (DIG)-labeled probe --- p.54 / Chapter 3.2.2 --- Probe specificity --- p.57 / Chapter 3.2.3 --- AS expression level under light/dark treatments by Northern Blot analysis --- p.58 / Chapter 3.2.4 --- AS expression level in young seedlings by Northern Blot analysis --- p.62 / Chapter 3.2.5 --- AS expression level in podding soybean by Northern Blot analysis --- p.64 / Chapter 3.3 --- Cloning of AS genes from high protein content soybeans --- p.66 / Chapter 3.3.1 --- "PCR amplification of AS1 and AS2 full length cDNA from the first-strand cDNA of high portein content cultivar soybean, YuDoul2" --- p.66 / Chapter 3.3.2 --- Nucleotide sequences analysis of AS1 and AS2 full-length cDNA clones --- p.68 / Chapter 3.4 --- Construction of AS1 and AS2 transgenic rice --- p.75 / Chapter 3.4.1 --- Construction of AS1 and AS2 constructs --- p.75 / Chapter 3.4.2 --- Transformation of chimeric gene constructs into Agrobacterium tumefaciens --- p.75 / Chapter 3.4.3 --- Agrobacterium mediated transformation into Oryza sativa calli to regenerate transgenic rice --- p.76 / Chapter 3.4.4 --- PCR screening of transgene from transgenic AS1 and AS2 rice --- p.76 / Chapter 3.4.5 --- Quantitative PCR analysis of the transgene expression --- p.81 / Chapter 3.4.6 --- Quantitative amino acid analysis in mature transgenic rice grain --- p.83 / Chapter 4 --- Discussion --- p.89 / Chapter 4.1 --- The role of asparagine and asparagine synthetase in nitrogen assimilation and sink-source relationship in soybean --- p.89 / Chapter 4.2 --- Comparative study of AS between different high seed protein content crops --- p.92 / Chapter 4.3 --- The attempt to find out the reason for the strong AS1 expression detected in high protein soybean cultivars --- p.92 / Chapter 4.4 --- Other factors affecting seed protein contents --- p.93 / Chapter 4.5 --- Rice seed quality improvement by nitrogen assimilation enhancement --- p.94 / Chapter 4.6 --- Comparative study of amino acid profile and seed total protein in other transgenic rice --- p.95 / Chapter 4.7 --- Possible reason of higher seed protein content in AS2 transgenic rice --- p.96 / Chapter 4.8 --- Selectable marker --- p.97 / Chapter 5 --- Conclusion and Prespectives --- p.99 / Chapter 6 --- References --- p.102 / Chapter 7 --- Appendix --- p.112 / Appendix I: Major chemicals and reagents used in this research --- p.112 / "Appendix II: Major buffer, solution and gel used in this research" --- p.114 / Appendix III: Commercial kits used in this research --- p.117 / Appendix IV: Major equipments and facilities used in this research --- p.118 / Appendix V: Primer list --- p.119

No / The influence of surfactants on the stability of cyclodextrin (CD) Pickering emulsions is not well understood. In this study, we report two-way effects of Tween 80 and soybean lecithin (PL) on the long term stability of Pickering emulsions stabilized by the self-assembled microcrystals of alpha-CD and medium chain triglycerides (MCT). The CD emulsions in the absence and presence of Tween 80 or PL at different concentrations were prepared and characterized by the droplet size, viscosity, contact angle, interfacial tension and residual emulsion values. After adding Tween 80 and PL, similar effects on the size distribution and contact angle were observed. However, changes of viscosity and interfacial tension were significantly different and two-way effects on the stability were found: (i) synergistic enhancement by Tween 80; (ii) inhibition at low and enhancement at high concentrations by PL. The stability enhancement of Tween 80 was due to the interfacial tension decrease caused by the interaction of Tween 80 with CD at the o/w interface at lower concentrations, and significant viscosity increase caused by the Tween 80-CD assembly in the continuous phase. For PL at low concentrations, the replacement of alpha-CD/MCT by alpha-CD/PL particles at the o/w interface was observed, leading to inhibitory effects. High concentrations of PL resulted in an extremely low interfacial tension and stable emulsion. In conclusion, the extensive inclusion of surfactants by CD leads to their unique effects on the stability of CD emulsions, for which the changes of viscosity and interfacial tension caused by host-guest interactions play important roles.

Conclusion. We observed a small but independent effect of soy intake and lipid lowering effect, even after taking into account the other important predicting factors - initial cholesterol, body composition, physical activity, dietary intake and age. The beneficial effect between soy protein intake and lipid profile were observed even with this relatively low level of soy protein consumption suggests that the effect of soy protein supplement use on lipid profile may be much greater than those observed here. The results of our study add to the existing evidence that soy protein may be beneficial in human lipid profile. Our data will be useful for planning effective education programs as well as providing background information for further interventional studies to prevent coronary heart disease. / Coronary Heart Disease (CHD) is the major cause of death in most developed countries and is rapidly increasing in developing countries. Recent studies showed that natural menopause confers a threefold increase in CHD risk. While many risk factors, such as hypertension, diabetes mellitus, obesity and physical inactivity contribute to the risk for CHD, lipid abnormalities are the major factor. Hyperlipidemia plays a central role in the atherosclerotic process. Recent studies showed that consuming soy, a food containing large amounts of soy protein, improves the plasma lipoprotein profile by decreasing total cholesterol, LDL cholesterol, triglycerides as well as increasing HDL level. Although soy is a main component of traditional Asian food, many of the studies on soy consumption have been conducted in Caucasian populations (table 1.2), among whom soy intake is rather low or almost nil, it was difficult to explore the association of soy protein intake and lipid profile in those populations. Soy products such as tofu and soymilk are traditional Chinese foods. With the changing dietary pattern, it gives rise to a range of intake from traditional to modern and increasing incidence of cardiovascular disease Hong Kong poses a unique opportunity for the investigation of the relation between soy protein intake and lipid profile. / For baseline age stratified subgroup analysis, our study results showed no association between soy protein intake and lipid pro file in women whose baseline age younger than 55.3 years old, but we did observe a positive association in women belonging to older subgroup. In the 12-month follow up analysis, for women whose baseline age was older than 55.3 years (mean age=58.4+/-2.1), after controlling for the potential confounders, soy protein intake was significantly associated with HDL cholesterol concentration (Linear Regression p=0.033, ANCOVA=0.011, P value for trend p=0.014), total cholesterol/HDL ratio (Linear Regression p=0.045) and LDL/HDL ratio (Linear Regression p=0.037). Similar observation was observed in the yearly change rate of HDL in 24-month follow up (Linear Regression p=0.047, P value for trend p=0.043). / For women whose initial cholesterol level was higher or equal to 200mg/dL, in our 2-year longitudinal analysis, after controlling for the potential confounders, soy protein intake was significantly associated with HDL (Linear Regression p=0.041) and cholesterol/HDL ratio (ANCOVA=0.022). We also observed a statistically significant trend for higher HDL cholesterol (p=0.038), with an increase of 11.4g in soy protein intake between the 1st and 3rd tertiles, our data showed a 3.8% increase in HDL. / In the 12-month longitudinal analyses, after controlling for the potential confounders, soy protein intake was significantly associated with HDL concentration (Linear Regression p=0.036). We also observed a statistically significant trend for higher HDL cholesterol (p=0.036), with an increase of 10.9g in soy protein intake between the 1st and 3rd tertiles, our data showed a 7.9% increase in HDL. / Methods. 307 women aged between 48 to 62 years were recruited from community subjects residing in housing estates in Shatin. Women within the first 12 years of menopause, with no history of malabsorption syndromes, chronic liver kidney diseases, parathyroid diseases, gastric operation or cancer and without currently taking lipid lowering therapy were included in the study. We estimated the dietary intake of soy foods and other key nutrients by using quantitative food frequency method. We recorded serum values of fasting cholesterol, LDL cholesterol, HDL cholesterol and triglycerides as well as other covariance measurement. Soy protein consumption was categorized as tertiles of intake and related to lipid profile. / Objectives. In order to study the relation between soy protein intake and lipid profile in the early postmenopausal Chinese women in Hong Kong, we conducted the study from February 2000 to February 2002, as a part of the population-based soy consumption and bone mineral density study. The hypothesis to be tested is that high intake of dietary soy protein has a beneficial effect on lipid profile in the early postmenopausal Chinese women in Hong Kong. / Results. In our cross-sectional analysis, our findings showed that habitual dietary soy protein intake had a weak but statistically significant correlation with triglyceride concentration (Linear Regression p=0.045, ANCOVA p=0.045 P value for trend p=0.023), and the soy protein beneficial effects were more pronounced in women whose % of total body fat were higher than 33.4%. After controlling for the potential confounders, soy protein intake was significantly associated with triglyceride concentration (Linear Regression p=0.048, P value for trend =0.021), the average decrease in triglycerides were 24.6% and 29.1 % in the 2nd and 3rd tertile compared with the 1st tertile respectively. / Lam Siu Hung. / "February 2006." / Adviser: Ho Suzanne Sutying. / Source: Dissertation Abstracts International, Volume: 67-11, Section: B, page: 6300. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 181-191). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Diss. (sammanfattning) Umeå : Univ., 2007. / Härtill 5 uppsatser.