Identification of Isoflavonoid Metabolites in Humans

Epidemiological studies have associated high soy intake with a lowered risk for certain hormone-dependent diseases, such as breast and prostate cancers, osteoporosis, and cardiovascular disease. Soy is a rich source of isoflavones, diphenolic plant compounds that have been shown to possess several biological activities. Soy is not part of the traditional Western diet, but many dietary supplements are commercially available in order to provide the proposed beneficial health effects of isoflavones without changing the original diet. These supplements are usually manufactured from extracts of soy or red clover, which is another important source of isoflavones. However, until recently, detailed studies of the metabolism of these compounds in humans have been lacking. The aim of this study was to identify urinary metabolites of isoflavones originating from soy or red clover using gas chromatography - mass spectrometry (GC-MS). To examine metabolism, soy and red clover supplementation studies with human volunteers were carried out. In addition, the metabolism of isoflavones was investigated in vitro by identification of metabolites formed during a 24-h fermentation of pure isoflavones with a human fecal inoculum. Qualitative methods for identification and analysis of isoflavone metabolites in urine and fecal fermentation samples by GC-MS were developed. Moreover, a detailed investigation of fragmentation of isoflavonoids in electron ionization mass spectrometry (EIMS) was carried out by means of synthetic reference compounds and deuterated trimethylsilyl derivatives. After isoflavone supplementation, 18 new metabolites of isoflavones were identified in human urine samples. The most abundant urinary metabolites of soy isoflavones daidzein, genistein, and glycitein were found to be the reduced metabolites, i.e. analogous isoflavanones, a-methyldeoxybenzoins, and isoflavans. Metabolites having additional hydroxyl and/or methoxy substituents, or their reduced analogs, were also identified. The main metabolites of red clover isoflavones formononetin and biochanin A were identified as daidzein and genistein. In addition, reduced and hydroxylated metabolites of formononetin and biochanin A were identified; however, they occurred at much lower levels in urine samples than daidzein or genistein or their reduced metabolites. The results of this study show that the metabolism of isoflavones is diverse. More studies are needed to determine whether the new isoflavonoid metabolites identified here have biological activities that contribute to the proposed beneficial effects of isoflavones on human health. Another task is to develop validated quantitative methods to determine the actual levels of isoflavones and their metabolites in biological matrices in order to assess the role of isoflavones in prevention of chronic diseases.

Two Complete Genome Sequences of Phasey Bean Mild Yellows Virus, a Novel Member of the Luteoviridae from Australia

We present here the complete genome sequences of a novel polerovirus from Trifolium subterraneum (subterranean clover) and Cicer arietinum (chickpea) and compare these to a partial viral genome sequence obtained from Macroptilium lathyroides (phasey bean). We propose the name phasey bean mild yellows virus for this novel polerovirus.

Synthesis of Reduced Metabolites of Isoflavonoids, and their Enantiomeric Forms

Isoflavonoids are naturally occurring plant derived biochemicals, which act as phytoalexins. Isoflavonoids are of interest due to their estrogenic and other potential physiological properties, particularly in mammals that typically consume isoflavonoid rich nutrients such as soy and red clover. The literature review of this thesis mainly focuses on the reduced metabolites of hydroxy and/or methoxy substituted isoflavones with four groups: isoflavan-4-ols, isoflav-3-enes, isoflavans and α-methyldeoxybenzoins (1,2-diarylpropan-1-ones), which are all reduced metabolites of food derived isoflavones in mammals. Related isoflavan-4-ones are briefly discussed. Results of an extensive survey of the literature concerning the synthesis of polyhydroxy- or methoxysubstituted isoflavonoids and especially asymmetric approaches are discussed. The experimental section describes new synthetic methods to prepare polyphenolic reduced isoflavonoid structures such as isoflav-3-enes, isoflavan-4-ones, cis- and trans-isoflavan-4-ols, 1,2-diarylpropan-1-ones and isoflavans by various hydride reagents and hydrogenations. The specific reactivity differences of various hydride reagents toward isoflavonoids are discussed. The first enantioselective synthesis of natural (S)-(-)-equol and the opposite enantiomer (R)-(+)-equol is also described by the asymmetric iridium PHOX catalysed hydrogenation of isoflav-3-enes. Both of these equol enantiomers are found to possess biological activity in mammals due to estrogen receptor binding activity. The natural enantiomer prefers estrogen receptor β and the R-enantiomer prefers the estrogen receptor α. Also the precursor, isoflav-3-ene, is found to possess positive biological effects on mammals. In connection with the synthetic work, the (S)-(-)-equol was discovered from serum of ewes after isoflavone rich red clover feeding. The chiral HPLC method was developed to identify natural equol enantiomer for the first time in this species. The first synthesis of natural isoflavonoid (R)-(-)-angolensin and its enantiomer (S)-(+)-angolensin is desribed by the use of recyclable chiral auxiliaries (chiral pseudoephedrines). The method offers a general approach also to other natural polyphenolic 1,2-diarylpropan-1-ones and to further study isoflavonoid metabolism in human and other mammals. The absolute configurations of these new chiral isoflavonoid metabolites were determined by X-ray spectroscopy. Also thorough NMR and MS analysis of synthesised structures are presented.

Vaporization and collision modeling of liquid fuel sprays in a co-axial fuel and air pre-mixer

Droplet collision occurs frequently in regions where the droplet number density is high. Even for Lean Premixed and Pre-vaporized (LPP) liquid sprays, the collision effects can be very high on the droplet size distributions, which will in turn affect the droplet vaporization process. Hence, in conjunction with vaporization modeling, collision modeling for such spray systems is also essential. The standard O'Rourke's collision model, usually implemented in CFD codes, tends to generate unphysical numerical artifact when simulations are performed on Cartesian grid and the results are not grid independent. Thus, a new collision modeling approach based on no-time-counter method (NTC) proposed by Schmidt and Rutland is implemented to replace O'Rourke's collision algorithm to solve a spray injection problem in a cylindrical coflow premixer. The so called ``four-leaf clover'' numerical artifacts are eliminated by the new collision algorithm and results from a diesel spray show very good grid independence. Next, the dispersion and vaporization processes for liquid fuel sprays are simulated in a coflow premixer. Two liquid fuels under investigation are jet-A and Rapeseed Methyl Esters (RME). Results show very good grid independence in terms of SMD distribution, droplet number distribution and fuel vapor mass flow rate. A baseline test is first established with a spray cone angle of 90 degrees and injection velocity of 3 m/s and jet-A achieves much better vaporization performance than RME due to its higher vapor pressure. To improve the vaporization performance for both fuels, a series of simulations have been done at several different combinations of spray cone angle and injection velocity. At relatively low spray cone angle and injection velocity, the collision effect on the average droplet size and the vaporization performance are very high due to relatively high coalescence rate induced by droplet collisions. Thus, at higher spray cone angle and injection velocity, the results expectedly show improvement in fuel vaporization performance since smaller droplet has a higher vaporization rate. The vaporization performance and the level of homogeneity of fuel-air mixture can be significantly improved when the dispersion level is high, which can be achieved by increasing the spray cone angle and injection velocity. (C) 2012 Elsevier Ltd. All rights reserved.

Evidence for X(5) critical point symmetry in 128Ce

Lifetimes of excited states in 128Ce were measured using the recoil distance Doppler-shift (RDDS) and the Doppler-shift attenuation (DSAM) methods. The experiments were performed at the Wright Nuclear Structure Laboratory of Yale University. Excited states of 128Ce were populated in the 100Mo(32Si,4n) reaction at 120 MeV and the nuclear γ decay was measured with an array of eight Clover detectors positioned at forward and backward angles. The deduced yrast transition strengths together with the energies of the levels within the ground-state (gs) band of 128Ce are in agreement with the predicted values for the X(5) critical point symmetry. Thus, we suggest 128Ce as a benchmark X(5) nucleus in the mass A ≈ 130 region. © World Scientific Publishing Company.

复合式高能γ探测器优化设计Monte Carlo模拟计算

用MonteCarlo方法系统模拟计算了以NaI晶体与BGO晶体为探测介质的clover���cluster复合式高能γ探测器的效率 .对于相同的介质几何 ,BGO复合式探测器的全能峰效率远高于相应的NaI复合式探测器 .用多块76× 1 2 7BGO晶体制作成的clover���cluster复合式高能γ探测器对 2 2 0MeV的γ光子的全能峰特征效率仍然高于 40 % ,绝对效率增加因子达 2 4与 2 7.

Experimental study of the beta-delayed neutron decay of N-21

The first spectroscopic study for the beta decay of N-21 is carried out based on beta-n, beta-gamma, and beta-n-gamma coincidence measurements. The neutron-rich N-21 nuclei are produced by the fragmentation of the E/A=68.8 MeV Mg-26 primary beam on a thick Be-9 target and are implanted into a thin plastic scintillator that also plays the role of beta detector. The time of flight of the emitted neutrons following the beta decay are measured by the surrounding neutron sphere and neutron wall arrays. In addition, four clover germanium detectors are used to detect the beta-delayed gamma rays. Thirteen new beta-delayed neutron groups are observed with a total branching ratio of 90.5 +/- 4.2%. The half-life for the beta decay of N-21 is determined to be 82.9 +/- 7.5 ms. The level scheme of O-21 is deduced up to about 9 MeV excitation energy. The experimental results for the beta decay of N-21 are compared to the shell-model calculations.

奇奇核170Re在束g谱学和176Ir EC/b+衰变研究

本论文在 166MeV 的束流能量下，利用重离子熔合蒸发反应 142 Nd（32 S， 1p3nγ ）170 Re 布居了 170 Re 的高自旋激发态，用 12 套带 BGO反康普顿抑制的高 纯锗探测器阵列进行了在束 X-γ 和γ γ − 符合测量。在 210MeV 的束流能量下， 利用重离子熔合蒸发反应 146 Nd（35 Cl，5nγ ）176 Ir 产生具有 β+ / EC 衰变性质的 核素 176 Ir, 由氦喷嘴快速带传输系统将反应产物送到低本底区，用一台小平面探 测器、一台轴对称型高纯锗探测器和一台 CLOVER 探测器进行了γ γ − 符合和 t γ − 单谱测量。基于这些测量结果，研究了 170 Re 的高自旋态和 176 Ir 的 β+ / EC 衰变性质。 本工作基于γ−γ符合关系及周围邻近核已知信息的综合分析，首次建立了包 括 16 个能级 26 条γ 跃迁的双奇核 170 Re 的转动带能级纲图，并利用推转壳模型、 粒子转子模型等对其进行了分析和讨论。根据实验提取出的准粒子顺排、 Routhian、旋称劈裂等结构信息和邻近双奇核带结构系统学知识的比较分析，指 出该带的两准粒子组态是 11/213/2 hi π ν ⊗ ，并且该带在低自旋出现旋称反转。 基于对 176 Ir 核 β+ / EC衰变实验数据的离线处理分析，对早先发表的 176 Ir 衰 变γ 跃迁进行确认的同时又发现了 4 个新能级和 13 条新的γ 射线，丰富了 176 Os 核的低位激发态能级纲图。根据典型γ 射线的衰变时间谱建议了 176 Ir核的一个长 寿命低自旋同核异能态，同时通过两准粒子耦合的半经验计算建议了其组态。

双奇核188Au的高自旋态能级结构研究

作为核物理的前沿领域之一，高自旋态的研究能够提供核结构、核形变、核子耦合等非常有价值的信息。本论文内容为研究188Au 高自旋态的结构特性。　　利用能量为86 和90MeV的19F束流，通过重离子熔合蒸发反应173Yb(19F,4nγ)研究了双奇核188Au 的高自旋态能级结构。利用GEMINI 探测器阵列进行了γ射线的激发函数、X-γ和γ-γ-t 符合测量，共获取了约160×106 个符合事件。基于实验测量结果，对原有的双奇核188Au 能级纲图做了较大的修改。新发现了两个转动带，分别对应建立在π1/2[541]⊗ν1/2[510] 和π1/2[541]⊗νi13/2 组态上的双退耦带和半退耦带。对前人所建立的正宇称带能级进行了修改，发现了一条新的20+态能级，并且指定其为πh11/2⊗2νi13/2h9/2的四准粒子结构。考查与费米面临近的单粒子轨道γ形变驱动效应，188Au 晕态带的γ∼70°非轴对称的形变可能是由h11/2− 奇质子驱动的。π1/2[541]⊗νi13/2 带观察到在低自旋发生旋称反转现象，对这个组态带的A∼180 区旋称反转系统性分析表明，反转点的转动惯量c Θ 函数不随NpNn而变，这与其它几个区有明显的不同。 另外，本论文还介绍了利用16 路Segment 型Clover 探测器的γ射线线性极化测量方法。并且利用60Co 源的两条级联γ射线对其灵敏度进行测试，发现能有效地测量γ射线的电磁属性，从而确定能级的宇称

17B的实验研究

核反应总截面是描述原子核反应基本特征的一个重要的物理量，从中可以得到有关核反应、核结构和核内核子分布的信息。利用透射法测量核的反应总截面对于模型没有依赖性，因而得到的结果更加可靠。在兰州放射性束流线（RIBLL）上利用透射法测量了17B与C、Pb靶的反应，得到了17B与C、Pb的反应总截面和去除双中子截面，同时首次尝试用CLOVER���测器测量与碎片符合的γ能谱。利用半经验的Shen公式可以很好地拟合稳定核的反应总截面的实验结果，但是并不能很好地描述17B的实验结果，说明17B具有奇异结构。采用描述不稳定核的多种密度分布形式代入零力程的Glauber模型计算17B与C的反应总截面，并与实验结果进行了比较。发现采用Skyrme－Hatree－Fock（SHF）、双Gauss、Gauss＋HO计算的密度分布形式都能够很好地拟合整个能区内的实验结果。通过各种密度分布形式提取了17B的核子密度分布，发现17B的中子密度分布有很大的弥散，并证实了17B是由核芯15B加两个价中子组成的晕结构。将Skyrme－Hatree－Fock（SHF）、双Gauss、Gauss＋HO密度分布形式，输入Glauber模型与实验结果比较，提取了17B的物质均方根半径，不同密度分布提取的均方根半径值在实验误差范围内一致，也进一步验证了17B的晕结构。采用Woods－Saxon势来描述17B核外的价中子在核芯中形成的势场，通过解薛定鄂方程来求解价中子处于特定轨道的波函数。假设17B核外的价中子处于纯的s分波和d分波都不能很好地和实验结果符合。所以认为核外的价中子应该是混合组态，通过拟合实验结果求解出s分波谱学因子的值，证明核外的价中子处在2s1/2轨道的几率更大

12N、12B、11C 在Si靶上反应总截面研究

核反应总截面是描述原子核反应基本特征的一个重要的物理量，从中可以得到有关核反应、核结构和核内核子分布的信息。在由放射性束所产生的奇异核结构与各种反应机制的研究中，反应总截面更是具有特殊的重要性。具有奇异核结构的核的一个典型物理现象就是其反应总截面要比稳定核大得多，I.Tanihata等人最早就是通过对放射性束流的相互作用截面的测量发现了具有奇异结构的核－中子晕核。由于反应截面的测量对探测器的要求不高，而且数据分析过程相对较为简单，因此反应截面的测量已经成为放射性束物理研究的一个非常重要的实验手段。目前，奇异核研究的重点在丰中子一侧，理论预言的很多中子晕核如8He、11Li、11、14Be、15、17、19C、等已经从实验上得到证实，而理论预言的质子晕核目前只有8B得到实验证实，其他的核实验数据较少或相互矛盾，无法作出准确的结论。12N就是其中之一，理论学家认为其具有质子晕结构；而实验数据主要集中在的反应截面测量上，实验数据较少又相互矛盾或不肯定，不能得出确切结论。因此，进一步开展研究是十分必要和有意义的。考虑到以上因素，充分利用兰州放射性束流线现有的实验条件，我们采用束流透射法测量了中能区多个能量点12N、11C、12B与Si靶的反应总截面和12N去质子截面。同时用CLOVER���测器测量与碎片符合的γ能谱。利用透射法测量核的反应总截面对于模型没有依赖性，因而得到的结果更加可靠。利用Shen公式和采用各种密度分布形式的零力程Glauber模型计算了12N、11C、12B的激发函数并与实验结果比较，提取了12N、11C、12B的密度分布与核半径，对12N去质子截面作了简单分析。使用Shen公式拟合实验数据时发现，Shen公式在整个能区范围内基本上可以很好的拟合12B、11C的反应数据，但Shen计算得到的12N的反应总截面与实验测量值有明显的差异，计算结果偏小。在使用Glauber模型拟合实验数据之前，分别使用有限力程和零力程Glauber模型拟合了12C＋12C的实验数据。在计算时，两种Glauber模型输入的密度分布采用Gauss密度分布形式。结果显示，使用零力程Glauber模型可以更好的拟合实验数据。分别使用高斯密度分布、HO密度分布、双参数费米密度分布、SHF模型计算得到的密度分布、基于少体模型的Gauss＋尾巴密度分布形式代入零力程Glauber模型拟合实验数据。对于12B、11C、12C，这些密度分布形式代入零力程Glauber模型计算得到的激发函数曲线在整个能区范围内都可以很好的拟合实验数据。但是对于12N，只有基于少体模型的Gauss＋尾巴密度分布形式代入零力程Glauber模型计算得到的激发函数曲线可以拟合实验结果。其他的密度分布形式计算得到的激发函数曲线明显低于实验数据。提取的12B、11C、12N均方根半径显示，12N的核半径明显大于其镜像核12B及其周围的核素。对12B、11C、12N的分析结果表明：12N的密度分布形式和稳定核有明显的不同，最后一个质子的密度分布具有很大的弥散，其可能具有奇异结构。对12N去质子截面的简单分析也表明了这一点

Geography and Genetics of Ecological Speciation in Pea Aphids

During ecological speciation, divergent natural selection drives evolution of ecological specialization and genetic differentiation of populations on alternate environments. Populations diverging onto the same alternate environments may be geographically widespread, so that divergence may occur at an array of locations simultaneously. Spatial variation in the process of divergence may produce a pattern of differences in divergence among locations called the Geographic Mosaic of Divergence. Diverging populations may vary in their degree of genetic differentiation and ecological specialization among locations. My dissertation examines the pattern and evolutionary processes of divergence in pea aphids (Acyrthosiphon pisum) on alfalfa (Medicago sativa) and clover (Trifolium pretense). In Chapter One, I examined differences among North American aphid populations in genetic differentiation at nuclear, sequence-based markers and in ecological specialization, measured as aphid fecundity on each host plant. In the East, aphids showed high host-plant associated ecological specialization and high genetic differentiation. In the West, aphids from clover were genetically indistinguishable from aphids on alfalfa, and aphids from clover were less specialized. Thus, the pattern of divergence differed among locations, suggesting a Geographic Mosaic of Divergence. In Chapter Two, I examined genomic heterogeneity in divergence in aphids on alfalfa and clover across North America using amplified fragment length polymorphisms (AFLPs). The degree of genetic differentiation varied greatly among markers, suggesting that divergent natural selection drives aphid divergence in all geographic locations. Three of the same genetic markers were identified as evolving under divergent selection in the eastern and western regions, and additional divergent markers were identified in the East. In Chapter Three, I investigated population structure of aphids in North America, France, and Sweden using AFLPs. Aphids on the same host plant were genetically similar across many parts of their range, so the evolution of host plant specialization does not appear to have occurred independently in every location. While aphids on alfalfa and clover were genetically differentiated in most locations, aphids from alfalfa and clover were genetically similar in both western North America and Sweden. High gene flow from alfalfa onto clover may constrain divergence in these locations.

Análisis y evaluación de pérdidas en la cosecha de trébol blanco (Trifolium repens L.) de dos sistemas de recolección : el tradicional y recolección directa

p.127-131

The ecology of Bacillus thuringiensis on the Phylloplane: colonization from soil, plasmid transfer, and interaction with larvae of Pieris brassicae

Seedlings of clover (Triflorium hybridum) were colonized by Bacillus thuringiensis when spores and seeds were co-inoculated into soil. Both a strain isolated in the vegetative form from the phylloplane of clover, 2810-S-4, and a laboratory strain, HD-1, were able to colonize clover to a density of about 1000 CFU/g leaf when seeds were sown in sterile soil and to a density of about 300 CFU/g leaf in nonsterile soil. A strain lacking the characteristic insecticidal crystal proteins produced a similar level of colonization over a 5-week period as the wild type strain, indicating that crystal production was not a mitigating factor during colonization. A small plasmid, pBC16, was transferred between strains of B. thuringiensis when donor and recipient strains were sprayed in vegetative form onto leaves of clover and pak choi (Brassica campestris var. chinensis). The rate of transfer was about 0.1 transconjugants/recipient and was dependent on the plant species. The levels of B. thuringiensis that naturally colonized leaves of pak choi produced negligible levels of mortality in third instar larvae of Pieris brassicae feeding on the plants. Considerable multiplication occurred in the excreted frass but not in the guts of living insects. Spores in the frass could be a source of recolonization from the soil and be transferred to other plants. These findings illustrate a possible cycle, not dependent on insect pathology, by which B. thuringiensis diversifies and maintains itself in nature.

Projecto de um sistema de rega para uma exploração agrícola na Beira Gomes

Trabalho de Projecto de Natureza Científica para obtenção do grau de Mestre em Engenharia Civil