Interactions of thiamine with anions: (Hthiamine)(thiamine) heptaiododimercurate dihydrate and its dimethanol monohydrate

In the title compounds, 3-[(4-amino-2-methyl-5-pyrimidinio)methyl] -5-(2-hydroxyethyl)-4-methylthiazolium(2+) 3-[(4-amino-2-mcthyl-5-pyrimidinyl)methyl]-5-(2-hydroxyethyl)-4-methylthiazolium( 1+) heptaiododimercurate dihydrate, (C12H18N4OS)(C12H17N4OS)[Hg2I7]. 2H(2)O, (I), and its dimethanol monohydrate, (C12H18N4OS)(C12H17N4OS)[Hg2I7]. 2CH(3)OH . H2O (2), a crystallographic centre of symmetry in (1) or a twofold axis in (2) is imposed between the protonated and deprotonated thiamine molecules, resulting in a statistically half-occupied proton attached at N1' of the pyrimidine ring. The Hg2I73- anion, residing on the centre of symmetry in (1) or on the twofold axis in (2), interacts with two thiamine molecules, each through a C2-H ... I ... pyrimidine-ring interaction. This bridging interaction is a characteristic of thiamine in the F conformation.

A NOVEL METAL THIAMINE COMPLEX WITH A CYCLIC DIMER FORMED BY METAL-BRIDGED 2 THIAMINE LIGANDS - CRYSTAL-STRUCTURE OF [MN(THIAMINE)CL2(H2O)]2[THIAMINE]2CL4.2H2O

The crystal structure of [Mn(thiamine)Cl2(H2O)]2[thiamine]2Cl4.2H2O has been determined by X-ray diffraction methods. The compound contains a cyclic dimer of a complex cation with two thiamine ligands bridged by two Mn(II) ions across a crystallographic center of symmetry. Each Mn(II) is coordinated by two chloride atoms, a water molecule, a N(1') atom of the pyrimidine from a thiamine and an O(53) atom of the hydroxyethyl side chain from another thiamine. There are two free-base thiamine molecules related by a center of symmetry in the unit cell, which form a base-pair through the hydrogen bonds. Both the independent thiamine molecules in the asymmetric unit assume the common F conformation with phi-T = 10.0(9) and 3.6(10) and phi-P = 85.6(7) and 79.6(7), respectively. The compound provides a possible model for a metal-bridged enzyme-coenzyme complex in thiamine catalysis. Crystallographic data: triclinic, space group P1BAR, a = 12.441(4), b = 13.572(4), c = 11.267(3) angstrom, alpha = 103.15(2), beta 89.03(3), gamma = 115.64(2)-degrees, Z = 1, D(calc) = 1.524 g cm-3, and R = 0.050 for 3019 observed reflections with I > 3-sigma(I).

青海糯性春小麦综合标记辅助育种研究

选用全糯小麦品种‘糯麦1号’与青海主要栽培品种‘阿勃’杂交，综合利用改良碘染色法、SDS-PAGE法和Waxy基因的分子标记等，对杂交后代进行了鉴定筛选。最终从F2代鉴定出了5粒全糯种子，从F_代鉴定出8株Wx-B1亚基缺失的植株。对全糯株系F_3代的农艺性状进行评价，5个全糯株系的综合农艺性状都优于‘糯麦1号’，与‘阿勃’较接近。测定全糯株系和Wx-B1亚基缺失植株F_4代种子的直链淀粉含量，5个全糯株系F_4代种子的直链淀粉含量接近于0，8个Wx-B1亚基缺失植株F_4代种子的直链淀粉含量在总 体上比‘阿勃’的直链淀粉含量低。研究表明，采用综合标记辅助选择可快速而准确地获得适合在青海栽培的全糯和部分糯性小麦。

弱筋小麦品种高分子量谷蛋白亚基组成分析

为了有效地利用从国内其他地区引进的低蛋白弱筋种质材料,采用SDS-PAGE技术对63份材料进行了高分子量谷蛋白亚基组成的分析.结果表明,参试材料中共有14种HMW-GS类型,Glu-A1位点上有Null、1和2~*三种类型,以Null为主(52.4%);Glu-B1位点上有13+16、17+18、7、7+8、7+8+9和7+9六种类型,以7+8为主(55.6%);Glu-D1位点上有10、12、2+12、5+10、5+12五种类型,以2+12为主(61.9%),而5+10为27%.亚基组合类型共有22种,以"1,7+8,2+12"为主(22.2%).品质评分频率最高的是8分,为38.1%,其次为6分,为14.3%,5分的为9.5%,但品质评分为10分的也有5个材料,频率为7.9%.优质亚基含量等同或高于其他同类研究,品质评分也相对较高,这说明我国弱筋小麦的选择,需要加强对HMW-GS组成的分析.

硬粒小麦-节节麦人工合成种HMW-GS组成及1.5＋10亚基的遗传分析

对CIMMYT的99份硬粒小麦-节节麦人工合成种（简称合成种）的HMW-GS组成分析发现，Glu-B1和Glu-D1位点的变异类型比普通小麦丰富，分别有9种和12种亚基类型；筛选出含有比5＋10亚基更优质的1．5＋10和5＋12亚基的合成种分别有8份和1份；含有优质亚基1．5＋10的合成种与普通小麦杂交结实正常；对2个合成种与2个普通小麦品种的8个止反交组合F1种子电泳发现，优质亚基1．5＋10在F1代能正常表达，双亲所有亚基任F1代都得到表达，表现共显性遗传。本研究为优质亚基1．5＋10和5＋12转育到普通小麦中奠定了基础。

青海高原春小麦HMW－GS等位基因变异及其对面团流变学特性的作用

为了给青海高原春小麦改良提供资料，分析了于海育成和国内外引进的125个春小麦品种（系）的HMW－GS等位基因变异，并测定了其中58个品种（系）在青海高原环境下的面团流变学特性。结果表明：（1）参试春小麦品种（系）的HMW－GS存在广泛的变异，Glu-A1位点出现3个等位基因，Glu-B1位点出现5个等位基因，Glu-D1位点出现4个等位基因；共出现了22种HMW－GS组合形式，其中1，7＋8，2＋12和N，7＋8，2＋12组合类型出现频率最高，并发现了个别罕见的变异类型，如2＋10，10。（2）亚基组合类型对沉淀值、形成时间、稳定时间、评价值等品质性状影响较大，对籽粒蛋白质含量、出粉率和面粉吸水率的影响不显著。试验还表明，HMW－GS组成与面粉的筋力有密切关系。

小麦体细胞无性系Glu-1FH基因突变体的遗传分析

对Glu - 1 基因的Glu - A1 和Glu - B1 位点都发生突变的1 个小麦体细胞无性系后代的高分子量谷蛋白亚基组成进行的分析结果, 进一步证实了体细胞无性系发生了基因突变, 且发现同源染色体的等位基因易发生相同突变, 突变体多是纯合基因型。认为这是体细胞无性系变异稳定快的遗传根源。

初论特提斯构造域一些大型-超大型金属矿床集中区聚矿构造

特提斯构造域是目前世界关注和研究的地学前沿科学问题之一。I．Metcalfe从构造演化角度将其划分为古特提斯、中特提斯及新特提斯，代表了从古生代特斯洋三次打开与闭合的构造过程。从聚矿构造类型看，一些大型-超大型金属矿床不但与其构造域中的大陆边缘的张裂-伸展变形过程有密切关系。而且与其打开的同时，澳大利亚陆块内前泥盆纪大洋盆地的闭合及造山运动有关。

大河边—新晃超大型重晶石矿床地球化学特征及形成的地质背景

本文从矿物岩石学、地球化学和沉积盆地分析角度，研究贵州天柱大河边－湖南贡溪两个超大型重晶石矿床中重晶石（矿）岩及其共生岩石的地球化学特征及构造地质背景。天柱－新晃－玉屏寒武纪热水沉积成矿盆地是发育在大陆斜坡上的断陷型热水沉积成矿盆地，由于同生断层作用将盆地切割成一系列次级盆地，大河边－碧林及龙背－铜盆盖三级热水沉积成矿盆地是大型重晶石矿床的构造定位空间。重晶石矿层主要赋存于下寒武统牛蹄塘组第一岩性段的黑色岩系中。 重晶石矿层是由海底低温热水同生沉积作用形成（105－192℃），古热水场的地球化学类型为硫酸盐型热水。硫酸盐型热水发生大规模同生沉积成岩成矿作用形成超大型重晶石矿床，重晶石矿层之上的黑色碳质粘土岩可能形成于封闭、还原、滞流的深水沉积环境，黑色碳质粘土岩构成矿层的封闭保存条件。

大型铜矿集区成矿聚集机制与隐伏矿床定位预测－以中条山矿集区和铜陵矿集区为例

本文在分析了大型铜矿聚区找矿预测研究的重要性、不利条件及突破途径、预测理论和探测技术的基础上，以中条山和铜陵两大铜矿聚集区为主要研究区域，通过研究中条山胡－篦型铜矿床和铜陵地区主要铜矿床的大地构造背景和矿床的地质地球化学特征，分析成矿机制和成矿控制因素，总结了成矿规律和找矿标志。应用这些研究成果，并根据找矿工作靶区内系统的地质调查、构造地球化学研究和三频激电测量结果，对中条山矿集区的秦家沟测区和老宝滩测区以及铜陵矿集区的凤凰山矿田进行了隐伏矿床定位预测。通过上述研究，主要取得了如下几项结论：（1）利用综合信息在生产矿山的周边及深部的开展找矿预测研究解决中条山和铜陵这种大型矿集区内矿山接替资源的最理想途径之一。（2）中条山矿聚集区的胡家峪－篦子沟型铜矿床是在地壳递进演化过程中经多次不同性质的地壳活动及相关成矿作用而形成的多因复成矿床，篦子沟组的黑色片岩、层间滑动构造、面理同斜褶皱－冲断构造、钠长石化、方柱石化及阳起石化、硅化、碳酸盐化等和特征的三频激电相对相位异常是胡篦型铜矿床的找矿标志。（3）在中条山秦家沟测区的A1区和老宝滩测区的B1区内存在隐伏的胡－篦型铜矿体（4）铜陵矿集区内的大规模铜聚集成矿作用主要与受控于基底断裂的燕山晚期大规模中浅成的岩浆活动所造成，成矿定位过程是受构造的、围岩的和岩体的多方面因素非线性耦合控制的复杂的动力学过程。（5）铜陵矿集区的凤凰山矿田的深部存在中型以上的矽卡岩－斑岩复合型铜金隐伏矿床。

Vitamina C, caratenóides, compostos fenólicos e atividade antioxidante em genótipos de banana.

A banana é um alimento altamente energético (cerca de 100 calorias por 100g de polpa), cujos hidratos de carbono (em torno de 22%) são facilmente assimiláveis. Contém vitaminas C, A, B1 e B2, e pequenas quantidades de vitamina D, e uma maior percentagem de potássio, fósforo, cálcio e ferro, quando comparada com a maçã ou laranja (DE MARTINI et al., 1990).

Caratterizzazione proteomica di fluidi e tessuti in diverse condizioni fisio-patologiche

This thesis has been focused on the proteomic characterization of human saliva from donors of different ages, starting from birth up to adult age, and pediatric brain tumor tissues. The first study has been performed in order to compare the acid-insoluble fraction of saliva from preterm with at-term newborns and adults and establish if differences exist. In the second study medulloblastoma and pilocytic astrocytoma pediatric brain tumor extracts have been compared. In both studies 2- DE analysis was coupled with high resolution tandem mass spectrometry (MS/MS). The proteomic characterization of the acid-insoluble fractions of saliva from preterm newborns allowed to integrate data previously obtained on the acid-soluble fraction by HPLC-electrospray ionization (ESI)-mass spectrometry (MS), and to evidence several differences between preterm newborns, at-term newborns and adults. Spots differentially expressed between the three groups, according to image analysis of the gels, were submitted to in-gel tryptic digestion and the peptide mixture analyzed by high performance HPLC-ESI-MS/MS for their characterization. By this strategy, we identified three over-expressed proteins in atterm newborns with respect to preterm newborns and adults (BPI fold-containing family A member 1, two proteoforms of annexin A1, and keratin type 1 cytoskeletal 13), and several over-expressed proteins in adults (fatty acid-binding protein, S100A6, S100A7, two proteoforms of S100A9, several proteoforms of prolactin-inducible protein, Ig kappa chain, two proteoforms of cystatin SN, one proteoform of cystatin S and several proteoforms of α-amylase 1). Moreover, for the first time, it was possible to assign by MS/MS four spots of human saliva 2-DE, already detected by other authors, to different proteoforms of S100A9. The strategy applied used a sequential staining protocol to the 2-DE gels, first with Pro-Q Diamond, that allows specific detection of phosphoproteins, and successively with total protein SYPRO Ruby stain. In the second study, proteomic analysis of two pediatric brain tumor tissues pointed out differences between medulloblastoma, the prevalent malignant tumor in childhood, and pilocytic astrocytoma, the most common, that only rarely shows a malignant progression. Due to the limited availability of bioptic tissue, the study was performed on pooled tumor tissues, and was focused on acid-insoluble fraction to integrate the characterization performed by a group of colleagues in Rome on the acid-soluble fraction by high performance HPLC-ESI-MS/MS. The results indicated that the two tumors exhibit different proteomic profiles and evidenced interesting differential expression of several proteins. Among them, peroxiredoxin- 1, peptidyl-prolyl cis–trans isomerase A, heterogeneous nuclear ribonucleoproteins A2/B1, mitochondrial isoform of malate dehydrogenase, nucleoside diphosphate kinase A, glutathione S-transferase P and fructose bisphosphate aldolase A resulted significantly over-expressed in medulloblastoma while glial fibrillary acidic protein, serotransferrin, α crystallin B chain, ferritin light chain, annexin A5, fatty acid-binding protein (brain), sorcin and apolipoprotein A-I resulted significantly over-expressed in pilocytic astrocytoma. In conclusion, the work done allowed to evidence the usefulness of using an integrated bottom-up/top-down approach, based on 2-DE-MS analysis and high performance MS in order to obtain a complete characterization of the proteome under investigation, revealing and identifying, not only peptides and small proteins, but also proteins with higher MW, that often it is not possible to identify by using exclusively a top-down ESI-MS approach.

Estudo da suscetibilidade aos antibióticos em isolados de Enterobacteriaceae provenientes de suiniculturas Portuguesas

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Caracterização da estrutura populacional de isolados de Escherichia coli provenientes de suiniculturas de produção intensiva e extensiva de Portugal

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Novel ingredients from brewers' spent grain - bioactivity in cell culture model systems and bioactivity retention in fortified food products

Functional food ingredients, with scientifically proven and validated bioactive effects, present an effective means of inferring physiological health benefits to consumers to reduce the risk of certain diseases. The search for novel bioactive compounds for incorporation into functional foods is particularly active, with brewers’ spent grain (BSG, a brewing industry co-product) representing a unique source of potentially bioactive compounds. The DNA protective, antioxidant and immunomodulatory effects of phenolic extracts from both pale (P1 - P4) and black (B1 – B4) BSG were examined. Black BSG extracts significantly (P < 0.05) protected against DNA damage induced by hydrogen peroxide (H2O2) and extracts with the highest total phenolic content (TPC) protected against 3-morpholinosydnonimine hydrochloride (SIN-1)-induced oxidative DNA damage, measured by the comet assay. Cellular antioxidant activity assays were used to measured antioxidant potential in the U937 cell line. Extracts P1 – P3 and B2 - B4 demonstrated significant (P < 0.05) antioxidant activity, measured by the superoxide dismutase (SOD) activity, catalase (CAT) activity and gluatathione (GSH) content assays. Phenolic extracts P2 and P3 from pale BSG possess anti-inflammatory activity measured in concanavalin-A (conA) stimulated Jurkat T cells by an enzyme-linked immunosorbent assay (ELISA); significantly (P < 0.05) reducing production of interleukin-2 (IL-2), interleukin-4 (IL-4, P2 only), interleukin-10 (IL-10) and interferon-γ (IFN-γ). Black BSG phenolic extracts did not exhibit anti-inflammatory effects in vitro. Hydroxycinnamic acids (HA) have previously been shown to be the phenolic acids present at highest concentration in BSG; therefore the HA profile of the phenolic extracts used in this research, the original barley (before brewing) and whole BSG was characterised and quantified using high performance liquid chromatography (HPLC). The concentration of HA present in the samples was in the order of ferulic acid (FA) > p-coumaric acid (p-CA) derivatives > FA derivatives > p-CA > caffeic acid (CA) > CA derivatives. Results suggested that brewing and roasting decreased the HA content. Protein hydrolysates from BSG were also screened for their antioxidant and anti-inflammatory potential. A total of 34 BSG protein samples were tested. Initial analyses of samples A – J found the protein samples did not exert DNA protective effects (except hydrolysate H) or antioxidant effects by the comet and SOD assays, respectively. Samples D, E, F and J selectively reduced IFN-γ production (P < 0.05) in Jurkat T cells, measured using enzyme linked immunosorbent assay (ELISA). Further testing of hydrolysates K – W, including fractionated hydrolysates with molecular weight < 3, < 5 and > 5 kDa, found that higher molecular weight (> 5 kDa) and unfractionated hydrolysates demonstrate greatest anti-inflammatory effects, while fractionated hydrolysates were also shown to have antioxidant activity, by the SOD activity assay. A commercially available yogurt drink (Actimel) and snack-bar and chocolate-drink formulations were fortified with the most bioactive phenolic and protein samples – P2, B2, W, W < 3 kDa, W < 5 kDa, W > 5 kDa. All fortified foods were subjected to a simulated gastrointestinal in vitro digestion procedure and bioactivity retention in the digestates was determined using the comet and ELISA assays. Yogurt fortified with B2 digestate significantly (P < 0.05) protected against H2O2-induced DNA damage in Caco-2 cells. Greatest immunomodulatory activity was demonstrated by the snack-bar formulation, significantly (P < 0.05) reducing IFN-γ production in con-A stimulated Jurkat T cells. Hydrolysate W significantly (P < 0.05) increased the IFN-γ reducing capacity of the snack-bar. Addition of fractionated hydrolysate W < 3 kDa and W < 5 kDa to yogurt also reduced IL-2 production to a greater extent than the unfortified yogurt (P < 0.05).