烟草FtsZ基因的表达特征分析及其对质体发生的影响

FtsZ (filamentation temperature-sensitive，fts)是广泛存在于原核生物和高等植物中的一种功能蛋白。它控制着原核细胞和质体的分裂过程。研究该基因的表达调控特征，为我们进一步认识原核细胞和质体分裂的分子机制及真核细胞的起源和进化等重要问题提供了新的思路。从烟草克隆FtsZ cDNA，构建了谷胱甘肽转移酶（GST, glutathione S-transferase，EC 2.5.1.18）与FtsZ融合蛋白的表达质粒，并将其导入到在异丙基-β-D-硫代半乳糖苷(IPTG)诱导下能高效表达的JM109大肠杆菌中。高表达的融合蛋白通过谷胱甘肽一琼脂糖(glutathione-agarose)亲和层析和SDS-聚丙烯酰胺凝胶电泳纯化后，用以免疫兔子制备抗血清。免疫印迹法表明烟草FtsZ基因表达具有明显的组织器官特征，在质体（叶绿体）分裂活跃部位表达强：幼嫩花瓣>幼叶>幼根>老叶>茎。黑暗处理l天对FtsZ表达似乎无影响，随黑暗培养时间延长，FtsZ蛋白表达逐渐降低，叶绿体转化成为数目众多（增加2-3倍）体积小的淡黄色或白色质体。该实验结果显示，光对植物FtsZ基因表达很可能无直接影响，FtsZ基因表达强弱是决定质体（叶绿体）分裂和细胞中质体（叶绿体）数目多少的主要原因之一。

Effect of Ru addition to CO/SiO2/HZSM-5 catalysts on Fischer-Tropsch synthesis of gasoline-range hydrocarbons

Microporous HZSM-5 zeolite and mesoporous SiO2 supported Ru-Co catalysts of various Ru adding amounts were prepared and evaluated for Fischer-Tropsch synthesis (FTS) of gasoline-range hydrocarbons (C-5-C-12). The tailor-made Ru-Co/SiO2/HZSM-5 catalysts possessed both micro- and mesopores, which accelerated hydrocracking/hydroisomerization of long-chain products and provided quick mass transfer channels respectively during FTS. In the same time. Ru increased Cor reduction degree by hydrogen spillover, thus CO conversion of 62.8% and gasoline-range hydrocarbon selectivity of 47%, including more than 14% isoparaffins, were achieved simultaneously when Ru content was optimized at 1 wt% in Ru-Co/SiO2/HZSM-5 catalyst.

Gasoline-range hydrocarbon synthesis over cobalt-based Fischer-Tropsch catalysts supported onSiO2/HZSM-5

Two types of SiO2 with different mesopore size and HZSM-5 zeolite were used to prepare hybrid supported cobalt-based catalysts. The textual and structural properties of the catalysts were studied using N-2 physisorption, X-ray diffraction (XRD), and H-2 temperature-programmed reduction (TPR) techniques. Fischer-Tropsch synthesis (FTS) performances of the catalysts were carried out in a fixed-bed reactor. The combination effects of the meso- and micropores of the supports as well as the interaction between supports and cobalt particles on FTS activity are discussed. The results indicate that the catalyst supported on the tailor-made SiO2 and HZSM-5 hybrid maintained both meso- and micropore pores during the preparation process without HZSM-5 particles agglomerating. The mesopores provided quick mass transfer channels, while the micropores contributed to high metal dispersion and accelerated hydrocracking/hydroisomerization reaction rate. High CO conversion of 83.9% and selectivity to gasoline-range hydrocarbons (C-5-C-12) of 55%, including more than 10% isoparaffins, were achieved simultaneously on this type of catalyst.

稀土与酸性磷（膦）酸酯固体配合物的合成、结构和振动光谱研究

本文合成了Ln(DMD)_3 (Ln = La, Pr, Sm, Eu, Ho, Yb, Lu, Y), Ln(DEP)_3 (Ln = La, Sm, Eu, Ho, Yb), Ln (DBP)_3 (Ln = La, Eu, Yb), Ln(BBP)_3 (Ln = La, Eu, Yb), Ln(DEHP)_3 (Ln = La, Ce,..., Lu, Y), Ln(EH[EHP])_3 (Ln = La, Ce,..., Lu, Y), 其中Ln(DMP)_3, Ln(DEP)_3, Ln (DBP)_3, Ln(BBP)_3为本文首次合成。培养出了未见文献报导的Pr(DMP)_3, La(DMP)_3·TMP配合物的单晶，晶体结构表明，Pr(DMP)_3中Pr原子的配位数为6， 配位多面体为稍扭曲的正八面体，配体以“O-P-O”桥和稀土离子配位而形成奇特的“双桥二十四环”的环套环的网状结构。La(DMP)_3·TMP中，La的配位数为7，配位多面体为扭曲的单帽三棱柱，配合物中中性磷酸酯TMP以P=0中的O与La配位，酸性配体以“O-P-O”桥的形式和La配位而形成奇特的“交替二—四桥”的链状结构。这两种配位方式及结构是文献中未报导过的。对配合物进行了热分析，根据热失重百分数并结合残渣的红外光谱证实，最终的分解产物为稀土偏酸盐。研究了配合物在4000-100cm~(-1)范围内的FTS-IR光谱和1500-50cm~(-1)范围内的拉曼光谱，对其主要的振动谱带进行了归属。其中150cm~(-1)附近的“稀土敏感带”指认为Ln-O配位键的伸缩振动带（νLn-O）。我们还指认了与配位密切相关的PO_2基团的ν_(as)PO_2带为1200-1136cm~(-1)，ν_sPO_2带为1126- 1064cm~(-1)。同时对Ln(DRP)_3配合物的ν_(as)PO_2(c), ν_sPO_2(c), νc-o-(p)异相，νc-o-(p)同相，νc-o-(o)，δPO_2等的振动带以及Ln(RRP)_3配合物中νp-o-(c), νp-c, νc-c-(o), νc-c-(p), δPO_2等的振动带进行了相应的指认。νLn-o频率较低，力常数（约为0.2mDyn/A）较小，所以Ln-o键基本上为离子键，配合物Δν值（Δν = ν_(as)PO_2 - ν_sPO_2）远小于Na盐中的Δν值，Ln-O键具有部分共价性质且随“镧系收缩”Ln-O键共价成分增大。通过对Ln(DRP)_3类配合物振动光谱的研究，Ln(DRP)_3配合物具有和Pr(DMP)_3单晶相似的配位形式和结构类型。Ln(RRP)_3的振动光谱和Ln(EH[EHP])_3的快原子轰击质谱结果, Ln(RRP)_3类配合物具有与Pr(DMP)_3相类似的确配位形式和结构类型。利用配合物的结构，对HDEHP和HEH[EHP]萃取稀土离子的萃取机理给予解释和说明。

基于多层抽样反馈的传感器网络时间同步算法

针对目前传感器网络时间同步算法存在的问题,提出了一种全网时间同步算法FTS(full-scale time synchronization),其基本思想是从整体角度对传感器网络实施逐轮次的推送式的时间同步操作,并通过少量抽样节点的反馈数据计算时间同步操作的有关参数。在常见的TelosB节点上给出了FTS算法的实现。实验和分析表明,FTS算法是一类收敛快速、资源高效、同步精度较高和运算复杂度较低的时间同步方法,目前已经在无线传感器网络测试平台上得到了应用。

一种手性性质随不同金属离子改变的共轭聚合物

化学传感器能识别一种特殊分子或一类分子的分子装置 ,基于共轭聚合物的化学传感器的设计原理一般都是利用其分子的电子效应和空间构象的变化 [1～ 4] ,而旋光分子的手性同样与其电子和空间结构相关 .本文试图建立一种依靠化合物手性 (如旋光度 )的变化检测金属离子的新方法 .由于含手性联萘结构的共轭聚合物具有较好的旋光热稳定性和高的旋光值 [5,6] ,以及聚合物主链上联吡啶基团可与许多过渡金属离子配位[4,7] ,我们设计合成了一种同时含有联吡啶和手性联萘基团的共轭聚合物 .结果表明 ,不同过渡金属离子与聚合物联吡啶单元的作用可引起聚合物吸收和发射光谱的变化 ,同时导致聚合物手性的变化 .1　实验部分1.1　仪器与试剂　 1 H NMR谱用 d-氯仿作溶剂 ,用四甲基硅烷作内标在 Varian3 0 0 MHz核磁仪上测定 ;红外光谱用 Bioard FTS-7傅里叶变换红外谱仪测定 ;凝胶凝透色谱 (GPC)用聚苯乙烯作标准 ,四氢呋喃作洗脱剂 ,在 Waters5 10仪器上测定 ;旋光度用 Perkin Elmer3 41LC型旋光仪测定 ;紫外 -可见光谱用 Varian Cray 5 0光谱仪测定 ;...

朝鲜淫羊藿中生物碱类新成分的分离提取及结构鉴定

淫羊藿系小檗科淫羊藿属植物 ,具有补肾壮阳、强筋骨、祛风湿之功效 .现代药理研究表明 ,它还具有防治骨质疏松和提高免疫功能等作用 .2 0 0 0年版中国药典收载了 5种淫羊藿属植物 ,朝鲜淫羊藿为其中一种 ,是长白山道地中药材之一 .朝鲜淫羊藿中化学成分的研究多集中在黄酮类化合物方面 [1～ 6 ] ,而对其生物碱类化合物的研究较少 [7,8] .本文对朝鲜淫羊藿中生物碱类化合物进行了研究 ,并发现了一种新的生物碱 (6-羟基 -1 1 ,1 2 -二甲氧基 -2 ,2 -二甲基 -1 ,8-二氧 -1 ,3 ,4,8-四氢 -2 H -7-氧杂 -2 -氮 -苯并 [c]菲 ) ,采用核磁共振波谱技术鉴定了该化合物的结构 ,并用电喷雾多级串联质谱对其结构的正确性进行了验证 .1　实验部分1 .1　仪器　APEX FT-ICRMS高分辨质谱仪 (德国 Bruker Daltonnics公司 ) ;FTS-7型红外光谱仪(美国 BIO-RAD公司 ) ;AV40 0核磁共振波谱仪 (德国 Bruker公司 ) ,以 CDCl3作溶剂及内标 ;X-4数字显示显微熔点仪 (北京泰克仪器有限公司 ) ;MSn在 ...

四[对-氧代(2-山萮酸乙酯基)]苯基卟啉银(Ⅱ)、镉(Ⅱ)的合成及其LB膜

Barraud等将具有两亲性的长链取代基接到苯环的对位上,制备了四[对-氧代(2-山?酸乙酯基)]苯基卟啉,具有较好的成膜性能,我们按其方法合成了两个高价过渡金属Ag(Ⅱ)、Cd(Ⅱ)的四[对-氧代(2-山?乙酯基)]苯基卟啉配合物,对它们的结构进行了表征,测定了π-A曲线及LB膜的可见吸收光谱和红外光谱。 所用试剂三氯甲烷、吡啶、乙醚、丙酸等使用前蒸馏,山?酸、醋酸银、醋酸镉均为分析纯。仪器为UV-3000型紫外可见分光光度计;FTS-20型红外光谱仪;JES-FE 3AS型顺磁共振

[Sm(CH_3CN)_9]·(AlCl_4)3·CH_3CN的合成和晶体结构

关于过渡金属乙腈配合物的合成和催化性能的研究在文献中已有很多报道。但是,含稀土元素的这类配合物的合成及催化性能的研究直到1986年才见报道。Thomas利用金属Eu和NOBF_4在乙腈中的反应制得了配合物[Eu(CH_3CN)_3(BF_4)_3]_x。本文报道了新乙睛稀土配合物[Sm(CH_3CN)_9]·(AlCl_4)_3·CH_3CN的合成及其晶体结构与分子结构。 (一) 实验 所有操作都在氩气保护下进行。无水SmCl_3用NH_4Cl法制得。AlCl_3经升华后使用。CH_3CN经P_2O_5干燥蒸馏。IR用Perkin-Elmer FTS-20红外光谱仪测定。

WASP-25b: a 0.6 MJ planet in the Southern hemisphere

We report the detection of a 0.6 MJ extrasolar planet by WASP-South, WASP-25b, transiting its solar-type host star every 3.76 d. A simultaneous analysis of the WASP, FTS and Euler photometry and CORALIE spectroscopy yields a planet of Rp= 1.22 RJ and Mp= 0.58 MJ around a slightly metal-poor solar-type host star, [Fe/H]=- 0.05 ± 0.10, of R*= 0.92 Rsun and M*= 1.00 Msun. WASP-25b is found to have a density of ?p= 0.32 ?J, a low value for a sub-Jupiter mass planet. We investigate the relationship of planetary radius to planetary equilibrium temperature and host star metallicity for transiting exoplanets with a similar mass to WASP-25b, finding that these two parameters explain the radii of most low-mass planets well.

An exploration of seniors’ motivation to use mobile brain-exercise software

This article reports on the development of an iPhone-based brain-exercise tool for seniors involving a series of focus groups (FGs) and field trials (FTs). Four FGs with 34 participants were conducted aimed at understanding the underlying motivational and de-motivational factors influencing seniors’ engagement with mobile brain-exercise software. As part of the FGs, participants had approximately 40 minutes hands-on experience with commercially available brain-exercise software. A content analysis was conducted on the data resulting in a ranking of 19 motivational factors, of which the top three were challenge, usefulness and familiarity and 15 de-motivational factors, of which the top-three were usability issues, poor communication and games that were too fast. Findings were used to inform the design of three prototype brain-exercise games for the iPhone contained within one overall application, named Brain jog. Subsequently, two FTs were conducted using Brain jog to investigate the part that time exposure has to play in shaping the factors influencing engagement. New factors arose with respect to the initial FGs including the motivational factor feedback and the de-motivational factor boring. The results of this research provide valuable guidelines for the design and evaluation of mobile brain-exercise software for seniors.

Nutrição clínica aplicada à alimentação escolar

Relatório de estágio de mestrado, Nutrição Clínica, Universidade de Lisboa, Faculdade de Medicina, 2015

Transcriptional regulation of the mannosyltransferase-encoding gene pigm in inherited glycosylphosphatidylinositol (GPI) deficiency

RESUMO:O glicosilfosfatidilinositol (GPI) é um complexo glicolipídico utlizado por dezenas de proteínas, o qual medeia a sua ancoragem à superfície da célula. Proteínas de superfície celular ancoradas a GPI apresentam várias funções essenciais para a manutenção celular. A deficiência na síntese de GPI é o que caracteriza principalmente a deficiência hereditária em GPI, um grupo de doenças autossómicas raras que resultam de mutações nos genes PIGA, PIGL, PIGM, PIGV, PIGN, PIGO e PIGT, os quais sao indispensáveis para a biossíntese do GPI. Uma mutação pontual no motivo rico em GC -270 no promotor de PIGM impede a ligação do factor de transcrição (FT) Sp1 à sua sequência de reconhecimento, impondo a compactação da cromatina, associada à hipoacetilação de histonas, e consequentemente, impedindo a transcrição de PIGM. Desta forma, a adição da primeira manose ao GPI é comprometida, a síntese de GPI diminui assim como as proteínas ligadas a GPI à superficie das células. Pacientes com Deficiência Hereditária em GPI-associada a PIGM apresentam trombose e epilesia, e ausência de hemólise intravascular e anemia, sendo que estas duas últimas características definem a Hemoglobinúria Paroxística Nocturna (HPN), uma doença rara causada por mutações no gene PIGA. Embora a mutação que causa IGD seja constitutiva e esteja presente em todos os tecidos, o grau de deficiência em GPI varia entre células do mesmo tecido e entre células de tecidos diferentes. Por exemplo nos granulócitos e linfócitos B a deficiência em GPI é muito acentuada mas nos linfócitos T, fibroblastos, plaquetas e eritrócitos é aproximadamente normal, daí a ausência de hemólise intravascular. Os eventos transcricionais que estão na base da expressão diferencial da âncora GPI nas células hematopoiéticas são desconhecidos e constituem o objectivo geral desta tese. Em primeiro lugar, os resultados demonstraram que os níveis de PIGM mRNA variam entre células primárias hematopoiéticas normais. Adicionalmente, a configuração dos nucleossomas no promotor de PIGM é mais compacta em células B do que em células eritróides e tal está correlacionado com os níveis de expressão de PIGM, isto é, inferior nas células B. A presença de vários motivos de ligação para o FT específico da linhagem megacariocítica-eritróide GATA-1 no promotor de PIGM sugeriu que GATA-1 desempenha um papel regulador na sua transcrição. Os resultados mostraram que muito possivelmente GATA-1 desempenha um papel repressor em vez de activador da expressão de PIGM. Resultados preliminares sugerem que KLF1, um factor de transcrição restritamente eritróide, regula a transcrição de PIGM independentemente do motivo -270GC. Em segundo lugar, a investigação do papel dos FTs Sp demonstrou que Sp1 medeia directamente a transcrição de PIGM em ambas as células B e eritróide. Curiosamente, ao contrário do que acontece nas células B, em que a transcrição de PIGM requer a ligação do FT geral Sp1 ao motivo -270GC, nas células eritróides Sp1 regula a transcrição de PIGM ao ligar-se a montante e não ao motivo -270GC. Para além disso, demonstrou-se que Sp2 não é um regulador directo da transcrição de PIGM quer nas células B quer nas células eritróides. Estes resultados explicam a ausência de hemólise intravascular nos doentes com IGD associada a PIGM, uma das principais características que define a HPN. Por último, resultados preliminares mostraram que a repressão da transcrição de PIGM devida à mutação patogénica -270C>G está associada com a diminuição da frequência de interacções genómicas em cis entre PIGM e os seus genes “vizinhos”, sugerindo adicionalmente que a regulação de PIGM e desses genes é partilhada. No seu conjunto, os resultados apresentados nesta tese contribuem para o conhecimento do controlo transcricional de um gene housekeeping, específico-detecido, por meio de FTs genéricos e específicos de linhagem.-------------ABSTRACTC: Glycosylphosphatidylinositol (GPI) is a complex glycolipid used by dozens of proteins for cell surface anchoring. GPI-anchored proteins have various functions that are essential for the cellular maintenance. Defective GPI biosynthesis is the hallmark of inherited GPI deficiency (IGD), a group of rare autosomal diseases caused by mutations in PIGA, PIGL, PIGM, PIGV, PIGN, PIGO and PIGT, all genes indispensable for GPI biosynthesis. A point mutation in the -270GC-rich box in the core promoter of PIGM disrupts binding of the transcription factor (TF) Sp1 to it, imposing nucleosome compaction associated with histone hypoacetylation, thus abrogating transcription of PIGM. As a consequence of PIGM transcriptional repression, addition of the first mannose residue onto the GPI core and thus GPI production are impaired; and expression of GPI-anchored proteins on the surface of cells is severely impaired. Patients with PIGM-associated IGD suffer from life-threatening thrombosis and epilepsy but not intravascular haemolysis and anaemia, two defining features of paroxysmal nocturnal haemoglobinuria (PNH), a rare disease caused by somatic mutations in PIGA. Although the disease-causing mutation in IGD is constitutional and present in all tissues, the degree of GPI deficiency is variable and differs between cells of the same and of different tissues. Accordingly, GPI deficiency is severe in granulocytes and B cells but mild in T cells, fibroblasts, platelets and erythrocytes, hence the lack of intravascular haemolysis.The transcriptional events underlying differential expression of GPI in the haematopoietic cells of PIG-M-associated IGD are not known and constitute the general aim of this thesis. Firstly, I found that PIGM mRNA levels are variable amongst normal primary haematopoietic cells. In addition, the nucleosome configuration in the promoter of PIGM is more compacted in B cells than in erythroid cells and this correlated with the levels of PIGM mRNA expression, i.e., lower in B cells. The presence of several binding sites for GATA-1, a mega-erythroid lineage-specific transcription factor (TF), at the PIGM promoter suggested that GATA-1 has a role on PIGM transcription. My results showed that GATA-1 in erythroid cells is most likely a repressor rather than an activator of PIGM expression. Preliminary data suggested that KLF1, an erythroid-specific TF, regulates PIGM transcription but independently of the -270GC motif. Secondly, investigation of the role of the Sp TFs showed that Sp1 directly mediates PIGM transcriptional regulation in both B and erythroid cells. However, unlike in B cells in which active PIGM transcription requires binding of the generic TF Sp1 to the -270GC-rich box, in erythroid cells, Sp1 regulates PIGM transcription by binding upstream of but not to the -270GC-rich motif. Additionally, I showed that Sp2 is not a direct regulator of PIGM transcription in B and erythroid cells. These findings explain lack of intravascular haemolysis in PIGM-associated IGD, a defining feature of PNH. Lastly, preliminary work shows that transcriptional repression of PIG-M by the pathogenic -270C>G mutation is associated with reduced frequency of in cis genomic interactions between PIGM and its neighbouring genes, suggesting a shared regulatory link between these genes and PIGM. Altogether, the results presented in this thesis provide novel insights into tissuespecific transcriptional control of a housekeeping gene by lineage-specific and generic TFs.