990 resultados para NETTRA-G1


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将固定相合成与“收敛 /发散”方法相结合 ,合成了第一、二代苯乙炔树枝状分子 .通过 Heck-Cassar-Sonogashira-Hagihara偶联反应 ,将其中心和末端分别修饰上供电子的氨基和拉电子的硝基 ,得到第一、二代中心为氨基、末端为硝基的苯乙炔树枝状分子 NH2 -G1 -( NO2 ) 2 和 NH2 -G2 -( NO2 ) 4.用傅里叶变换红外光谱跟踪了整个固定相合成过程 .苯乙炔树枝状分子的紫外 -可见吸收光谱呈现出规律性变化

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利用ESI/MS~n、LC/MS、MALDI-TOF/MS等技术,从人参四逆汤水煎液中分析和鉴定了10种二萜生物碱:苯甲酰中乌头碱benzoylmesaconitine、苯甲酰乌头碱henzoylaconitine、苯甲酰次乌头碱benzoylhypacoitine、尼奥灵neoline、附子灵fuziline、14-乙酰基塔拉地萨敏14-acetyl-talatisamine、苯甲酰次乌头碱亚油酸酯14-benzoylhypaconine-8-linoleate、苯甲酰去氧乌头碱亚油酸酯14-benzoydeoxyaconine-8-oleate、苯甲酰次乌头碱棕榈酸酯14-benzoylhypaconine-8-palmitate、塔拉地萨敏talatisamine和人参皂甙R_(a1)、R_(a2)、R_(b1)、R_(b2)、R_(b3)、R_c、R_d、R_e、R_(g1)、R_(g2)、R_(g3)、R_f等12种人参皂甙。脂肪酸酯型生物碱和苯甲酰类生物碱的提取组分有抗失血性休克作用和弱的正性肌力作用。人参皂甙的提取组分具有明显的负性肌力作用。

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Dynamic mechanical properties of sulfonated butyl rubber ionomers neutralized with different amine or metallic ion (zinc or barium) and their blends with polypropylene (PP), high-density polyethylene (HDPE), or styrene-butadiene-styrene (SBS) triblock copolymer were studied using viscoelastometry. The results showed that glass transition temperatures of ion pair-containing matrix and ionic domains (T-g1 and T-g2, respectively) of amine-neutralized ionomers were lower than those of ionomers neutralized with metallic ions, and the temperature range of the rubbery plateau on the storage modulus plot for amine-neutralized ionomers was narrower. The modulus of the rubbery plateau for amine-neutralized ionomers was lower than that of ionomers neutralized with zinc or barium ion. With increasing size of the amine, the temperature range for the rubbery plateau decreased, and the height of the loss peak at higher temperature increased. Dynamic mechanical properties of blends of the zinc ionomer with PP or HDPE showed that, with decreasing ionomer content, the T-m of PP or HDPE increased and T-g1 decreased, whereas T-g2 or the upper loss peak temperature changed only slightly. The T-g1 for the blend with SBS also decreased with decreasing ionomer content. The decrease of T-g1 is attributed to the enhanced compatibilization of the matrix of the ionomer-containing ion pairs with amorphous regions of PP or HDPE or the continuous phase of SBS due to the formation of thermoplastic interpenetrating polymer networks by ionic domains and crystalline or glassy domains.

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The morphology and dynamic mechanical properties of blends of poly(ether imide) (PEI) and nylon 66 over the full composition range have been investigated. Torque changes during mixing were also measured. Lower torque values than those calculated by the log-additivity rule were obtained, resulting from the slip at the interface due to low interaction between the components. The particle size of the dispersed phase and morphology of the blends were examined by scanning electron microscopy. The composition of each phase was calculated. The blends of PEI and nylon 66 showed phase-separated structures with small spherical domains of 0.3 similar to 0.7 mu m. The glass transition temperatures (T(g)s) of the blends were shifted inward, compared with those of the homopolymers, which implied that the blends were partially miscible over a range of compositions. T-g1, corresponding to PEI-rich phase, was less affected by composition than T-g2, corresponding to nylon 66-rich phase. This indicated that the fraction of PEI mixed into nylon 66-rich phase increased with decreasing PEI content and that nylon 66 was rarely mixed into the PEI-rich phase. The effect of composition on the secondary relaxations was examined. Both T-beta, corresponding to the motion of amide groups in nylon 66, and T-gamma, corresponding to that of ether groups in PEI, were shifted to higher temperature, probably because of the formation of intermolecular interactions between the components.

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对四种大型海藻:江篱(Gracilaria textorii)、孔石莼(Ulva pertusa)、海带苗(Laminaria japonica)和多管藻(Polysiphonia urceolata)表面附着弧菌的多样性进行了分析,同时对多样性分析中采用的不同方法进行了比较。 利用TCBS培养基从四种海藻表面总共分离到12株细菌:G1、G2分离自江篱,U3分离自孔石莼,L4、L5、L6分离自海带苗,P7、P8、P9、P10、P11、P12分离自多管藻。菌株G1,G2属于盐单胞菌(Halomonas),其余10株细菌都属于弧菌(Vibrio)。 对12株细菌的酶学活性、抗生素抗性进行了研究。发现除G1、G2外,其余菌株都可产生淀粉酶和明胶酶。菌株G1、G2和U3对氨苄青霉素、链霉素和四环素有很强抗性。 采用多种分子生物学方法(16S rDNA、gyrB、RFLP、DGGE )分析了上述12株菌的系统发育关系,对不同方法获得的结果进行了比较。 12株菌的16S rDNA系统发生树可分为4个明显分支。G1、G2与细菌H. meridiana构成一个分支。U3与V. lentus构成一个分支。L4、L5、L6、P9、P10、P11、P12与V. tasmaniesis形成一个分支。P7、P8与V. splendidus形成一个分支。 以gyrB基因序列为基础构建的系统发生树将G1、G2以外的10株弧菌分成4个较大分支,L4、P9属于同一分支,L6、P8、P10、P11和P12属于一个大的分支, U3、L5各自构成一个分支。菌株G1、G2没有得到gyrB基因序列扩增带。表明试验设计的引物是弧菌特异性引物。相对16S rDNA,不同菌株间gyrB基因序列差异性更大。 RFLP分析结果显示,12株细菌HinfI酶切后得到6种带型,SmaI酶切后得到4种带型。结合16S rDNA、gyrB基因的分析结果可知,RFLP可以在种的水平上有效进行细菌多样性分析。 12株细菌的DGGE分析结果显示,G1、G2与其它菌株电泳图谱有明显不同。菌株U3独自构成一种带型,L5、L6构成一种带型。表明DGGE技术在属的水平上分析细菌多样性效果较好,在种的水平上分析细菌多样性有一定的局限性。

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中华哲水蚤是黄、东海浮游动物关键种,连接初级生产与较高营养级,在海洋生态系统中占据核心地位。在以往研究基础上,依据2006年3月至2007年8月黄海9个航次、胶州湾11个航次现场调查数据以及大量室内实验数据,本文研究了中华哲水蚤的繁殖、种群补充以及油脂积累的季节变化,期望初步阐明中华哲水蚤在黄海的种群动态及驱动因素、并构建其生活史概念模型。 3、4月份,黄海与胶州湾中华哲水蚤的生殖腺普遍发育成熟、产卵率较高,中华哲水蚤种群进入繁殖、种群补充的活跃期,恰好可以利用此时较好的食物条件。5、6月份,中华哲水蚤在黄海近岸海区仍表现出较高的产卵率,而陆架海区的中华哲水蚤生殖腺成熟度、产卵率逐渐降低,种群逐渐表现出度夏特征—C5占优势、积累大量脂类。整个夏季,中华哲水蚤的繁殖率、种群补充率在整个黄海都较低,尤其在黄海冷水团区:中华哲水蚤生殖腺停滞于未成熟的GS1-GS2期,产卵率为零。10月,中华哲水蚤在冷水团边缘潮汐锋区的繁殖、种群补充活跃,或许可以解释秋、冬季近岸种群的恢复。11月之后,随着垂直混合的加强,黄海冷水团逐渐消失,此时陆架区的中华哲水蚤种群结束度夏过程,并以较低的繁殖率进行种群补充,直到次年3、4月份食物环境转好,中华哲水蚤种群出现新一轮繁殖、补充高峰。本文依据中华哲水蚤雌体密度、产卵率、孵化率等计算了中华哲水蚤的种群潜在补充率,我们发现,陆架区种群仅在3、4月份出现较强补充以及冬季的微弱补充,而近岸海区除夏季外,于各个季节均可进行比较活跃的种群补充,其中春季种群补充规模、强度最大;整体来看,春季是黄海中华哲水蚤种群补充的最重要时期。 本文详细研究了中华哲水蚤的繁殖与各种环境、雌体自身因素之间的关系:1、统计结果表明,中华哲水蚤的产卵率与海区食物条件(浮游植物生物量、纤毛虫丰度)之间具有紧密联系,而与油囊体积无关;在饥饿培养条件下,中华哲水蚤只能维持3-6天的产卵,而添加食物2-7天内,即可逐渐恢复产卵;体内积累油脂在饥饿过程中有所消耗,不过仍然不能阻止产卵率的下降;这些结果表明中华哲水蚤繁殖的能量所需主要来自于近期摄食,而体内储存脂类可能主要用于代谢所需,是应对不利条件的一种能量缓冲。2、统计分析表明:产卵率与雌体前体长间具有一定的正相关性,这可能由于体长较长的雌体往往具有较高的怀卵量;产卵率与温度之间没有显著的统计关系,温度可能通过调节中华哲水蚤的代谢率、体长等间接影响到产卵率。3、值得一提的是,本文发现中华哲水蚤的产卵率与其生殖腺成熟度之间具有非常紧密的统计关系,这为通过保存样品估算产卵率以及长期历史样品、数据间的比较提供了可能。 春季硅藻水华是黄海及其邻近海域的重要季节性特征。本文针对硅藻水华与中华哲水蚤繁殖的关系进行了多次现场研究,我们发现,硅藻水华因其发生海区、硅藻优势种类不同而对中华哲水蚤繁殖具有复杂多样的影响。2006年3月,胶州湾东部海区发生中肋骨条藻(Skeletonema costatum)水华,中华哲水蚤的产卵率显著增高,但其孵化率相对较低(50%左右);2007年2月,胶州湾东部发生环纹劳德藻(Landeria annulata)水华,中华哲水蚤卵的孵化率在各种处理下均接近100%;2006年4月,黄海东北部发生太平洋海链藻(Thalassiosira pacifica)水华,产卵率与孵化率均处于调查海区的中等水平;2007年4月,黄海中部海区发生中肋骨条藻水华,中华哲水蚤在此次调查海区平均产卵率高达27.8 卵/雌体/天,远远高于以往同时期的调查结果,孵化率、幼体存活率也普遍较高;此次硅藻水华对于黄海陆架区中华哲水蚤的种群补充具有积极的促进作用。综上,硅藻水华对中华哲水蚤繁殖的影响表现出种类、海区特异性,其具体机理仍需进一步研究。 本文研究了黄海中华哲水蚤C5期油脂积累的区域、季节变化,并探讨了油脂积累对中华哲水蚤的生理、生活史的可能作用。黄海近岸区的C5期油囊体积常年较小,而陆架区则表现出明显的季节差异。在陆架区,C5期是中华哲水蚤度夏种群的主要组成部分,其油囊体积与度夏过程有密切联系:最大油囊体积(可占前体部体积的30%以上)出现于度夏准备期(5、6月),随着度夏过程的进行,油囊体积因代谢而不断消耗,至12月时,油囊体积降低到与近岸种群无异。次年4月以后,油脂积累可能随食物条件转好而再次开始积累。我们认为黄海中华哲水蚤所积累油脂除了可以为休眠提供能量以克服较长时期食物缺乏之外,还可能是休眠的诱导因素,对其生活史具有重要意义。 黄海近岸海区与陆架海区在水团特征、中华哲水蚤种群丰度与结构、繁殖特征、种群补充、油脂储存策略等方面具有显著差异,因此,本文分别讨论了这两个区域中华哲水蚤的生活史。黄海陆架海区中华哲水蚤在全年共有4-5个世代:11月末,中华哲水蚤结束度夏过程,种群中占优势的C5期个体开始蜕皮为成体、逐渐成熟、繁殖,由此产下的子代可称之为G0,此世代在食物条件较差的冬季发育成熟后可以产下G1;G1于3月初发育成熟,恰逢春季较好的食物条件,于是在3月初-5月中旬很可能发育G2、G3两个世代;假若陆架区食物环境适宜,5月中旬后很可能产生新的世代—G4;G3与G4两个世代的混合种群共同进入度夏过程,期间,一小部分C5蜕皮为成体,大部分C5保持滞育状态;至12月冷水团消退后,G3、G4世代C5蜕皮、成熟,开始新的生活周期。中华哲水蚤生殖腺成熟度、油囊体积的季节变化与上述概念模型具有较好的对应关系,另外,这两个参数简单易用、对环境变化敏感,可以尝试应用于气候变化与桡足类关系等长期研究中。 以往的研究更多的关注于黄海陆架区的中华哲水蚤种群,对近岸种群的关注较少,本文初步探讨了近岸种群的种群补充、世代情况。相对于陆架区,黄海近岸区中华哲水蚤的生活史更为复杂,区域差异较大。近岸海区中华哲水蚤产卵率、种群周转率较高;同时,近岸海区是多种鱼类的产卵场、索饵场,该区域的各营养级间的能量流动对黄海海洋生态系统的结构功能至关重要。因此,在将来的研究中,近岸海区中华哲水蚤种群的生态学、生活史应该受到同样的重视。

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A 21-d laboratory experiment was conducted to study, the phosphorus (P) utilization of two different diets by redlip mullet Liza haematocheila T. & S. Sand-filtered water in salinity 30 and temperature 25 degrees C was used. Twenty-nine fish individuals were divided into three groups: 11 to group 1 (G1) fed on diet 1, 11 to group 2 (G2) fed on diet 2, and 7 to contrast group. Diet 1 was a commercial feed, more valuable in nutrition than diet 2 that similar to natural detritus. The results show the intake phosphorus (IP) of G1 was significantly higher than that of G2, and both increased linearly with body size at a certain amount of diet. The retention phosphorus (RP) in fish of G1 was lower than G2. The relationship between retention phosphorus and body size was positive and stronger in G2. Significant difference in faecal phosphorus (FP) was found between G1 and G2. Body size significantly impacted the excretion phosphorus (EP) in G1 but G2. The loss of intake phosphor-us in G1 was 10.83-20.27 mg per g fish weight gain, higher than that in G2 for 6.63-9.56. Of the phosphor-us, about 10% was allocated into growth, 50% in faeces, and the rest lost in excretion. The main part of phosphorus was lost in faeces but excretion. The phosphorus budget of the fish could be described as 100IP = 7.40RP + 47.39FP + 36.63EP (Diet 1) or 100IP = 11.93RP + 56.64FP + 21.76EP (Diet 2).

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全球变化的研究发现:19世纪以来全球平均气温大约上升了0.6℃,并预计在21世纪会升高更多。温室效应的直接结果是导致全球地面增温,温度的升高会增加下垫面的蒸发量。根据全球水平衡原理,蒸发量应该与降水量相等,因而降水量也会相应增加。气候变化的情景模式研究表明,世界各地雨量变异大,在高纬度地区,包括北欧地区,降水会增加。 流域是水生系统的一个重要的汞和甲基汞的输入源。欧洲淡水系统中,鱼体汞含量过高的湖泊主要局限于北方的高纬度森林地区。在斯堪的纳维亚半岛,就有上万湖泊中的鱼体汞含量超过了健康食用标准0.5 mg/kg。水生系统甲基汞输入源包括:水体或沉积物通过甲基化形成甲基汞,通过大气干、湿沉降输入的甲基汞和陆地生态系统的地表径流输入的甲基汞。 本研究的主要目标是搞清楚未来气候变化引起的大气降水量的增加对北方高纬度森林流域土壤中汞的迁移和形态转化有何影响。未来大气降水量的增加可能会增强森林地区流域土壤中汞的迁移,进而对水生系统的汞循环产生影响。一方面,如果地下水水位上涨,当水流过富含有机质结合含汞土壤层时,将会增加土壤中汞和甲基汞的迁移性,由此而导致流域向河流、湖泊等水生系统中汞输出通量的增加。另一方面,氧化还原电位改变、DOC和营养物质的释放会加剧汞在土壤中的释放过程,并且可能会促进汞的形态转化,可能增加毒性较强的甲基汞的产量,最终导致甲基汞从土壤向水生系统的输出通量的增加。 本研究选择瑞典西海岸花园湖(Gårdsjön)的G1流域作为研究对象,在流域内进行人工模拟增加降雨试验,模拟未来气候变化降水量增加的情景,观测流域地表径流中汞和甲基汞的变化。并在流域内选择四个点(1×1m²),进行汞同位素加入示踪试验。测定土壤剖面甲基汞和总汞及其同位素组成,来分析汞的迁移和形态转化。主要研究结果如下: 1. 通过对G1流域土壤中甲基汞分布的调查,发现甲基汞含量范围为0.020-3.1ng/g,甲基汞含量与TOC含量具有正相关的关系,表明土壤甲基汞的迁移受TOC迁移的影响。估算G1流域的甲基汞储库通量为123g/km2,G1流域甲基汞储库容为0.65g。 2. 通过甲基汞含量和甲基汞储库在土壤剖面的分布,可以看出表层腐殖质层甲基汞浓度最高,甲基汞最富集,而在较深的土壤层位甲基汞储库容量最大,表明甲基汞具有向下迁移的趋势。 3. 根据总汞同位素组成结果,汞同位素(198Hg(NO3)2)加入土壤中后,202Hg/198Hg比值迅速下降,然后随时间推移而上升,表明加入土壤中的汞与土壤中原始汞形态不同,具有不同的迁移方式,且加入土壤中二价汞的迁移性比土壤中的原始汞更强。计算表明,外界加入的汞,将有50-60%长期滞留在土壤中。 4. 根据甲基汞的同位素组成结果,Wet点的新加入的198Hg甲基汞分数要大于其它三点,表明湿润土壤中汞甲基化速率要快于较干燥的土壤中。另外,Wet点的甲基汞分数比以前无人工模拟增加降雨的同位素示踪结果高很多,表明降水量增加会导致汞的甲基化作用在土壤中的形态转化过程中占主导地位,最终导致甲基汞产率的提高。 5. 人工模拟增加降雨试验进行后,地表径流中总汞的浓度没有发生变化,而甲基汞的浓度增大了很多,从0.03ng/L增大到了1ng/L以上,表明降水的增加可以促进土壤中汞的甲基化作用。 6. 地表径流向流域外总汞和甲基汞的输出量的计算显示,受人工模拟增加降雨的影响,流域地表径流量也会增加,最终年地表径流中总汞和甲基汞的输出通量都变大。

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The aim of the present study is twofold. Firstly, the paper investigates the undrained cyclic and post-cyclic behaviour of two silica sands by means of multi-stage cyclic triaxial tests. Secondly, based on the post-cyclic response observed in the element test, the authors formulate a simplified stress–strain relationship that can be conveniently used for the construction of p–y curves for liquefiable soils. The multi-stage loading condition consists of an initial cyclic loading applied to cause liquefaction, followed by undrained monotonic loading that aimed to investigate the post-cyclic response of the liquefied sample. It was found that due to the tendency of the liquefied soil to dilate upon undrained shearing, the post-liquefaction strain–stress response was characterised by a distinct strain–hardening behaviour. The latter is idealized by means of a bi-linear stress–strain model, which can be conveniently formulated in terms of three parameters, i.e.: (i) take-off shear strain, γto, i.e. shear strain required to mobilize 1 kPa of shear strength; (b) initial secant shear modulus, G1, defined as 1/γto; (c) post-liquefied shear modulus at large strain, G2 (γ⪢γto). Based on the experimental results, it is concluded that these parameters are strongly influenced by the initial relative density of the sample, whereby γto decreases with increasing relative density. Differently both shear moduli (G1 and G2) increases with increasing relative density. Lastly, the construction of new p–y curves for liquefiable soils based on the idealized bi-linear model is described.

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Dissertação apresentada à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Psicologia, com especialização em Psicologia Social e das Organizações.

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The Silurian-Devonian Galway Granite Complex (GGC ~425-380Ma) is defined here as a suite of granitoid plutons that comprise the Main Galway Granite Batholith and the Earlier Plutons. The Main Batholith is a composite of the Carna Pluton in the west and the Kilkieran Pluton in the east and extends from Galway City ~130km to the west. The Earlier Plutons are spatially, temporally and structurally distinct, situated northwest of the Main Batholith and include the Roundstone, Omey, Inis and Letterfrack Plutons. The majority of isotopic and structural data currently available pertain to the Kilkieran Pluton, several tectonic models have already been devised for this part of the complex. These relate emplacement of the Kilkieran Pluton to extension across a large east-west Caledonian lineament, i.e. the Skird Rocks Fault, during late Caledonian transtension. No chronological data have been published that directly and accurately date the emplacement of the Carna Pluton or any of the Earlier Plutons. There is also a lack of data pertaining to the internal structure of these intrusions. Accordingly, no previous study has established the mechanisms of emplacement for the Earlier Plutons and only limited work is available for the Carna Pluton. As a consequense of this, constituents of the GGC have not previously been placed in a context relative to each other or to regional scale Silurio-Devonian kinematics. The current work focuses on the Omey, Roundstone and Carna Plutons. Here, results of detailed field and Anisotropy of Magnetic Susceptibiliy (AMS) fabric studies are presented. This work is complemented by geological mapping that focuses on fault dynamics and contact relationships. Interpretation of AMS data is aided by rock magnetic experiment data and petrographic microstructural evaluations of representative samples. A new geological map of the the Omey Pluton demonstrates that this intrusion has a defined roof and base which are gently inclined parallel to the fold hinge of the Connemara Antiform. AMS and petrographic data show the intrusion is cross cut by NNW-SSE shear zones that extend into the country rock. These pre-date and were active during magma emplacement. It is proposed that the Omey pluton was emplaced as a discordant phacolith. Pre-existing subvertical D5 faults in the host rock were reactived during emplacement, due to regional sinistral transpression, and served as centralised ascent conduits. A central portion of the Roundstone Pluton was mapped in detail for the first time. Two facies are identified, G1 forms the majority of the pluton and coeval G2 sheets cross cut G1 at the core of the pluton. NNW-SSE D5 faults mapped in the country rock extend across the pluton. These share a geometrical relationship with the distribution of submagmatic strain in the pluton and parallel the majoity of mapped subvertical G2 dykes. These data indicate that magma ascent was controlled by NNW-SSE conduits that are inherently related to those identifed in the Omey Pluton. It is proposed that the Roundstone Pluton is a punched laccolith, the symmetry and structure of which was controlled by pre-exising host rock structures and regional sinistral transpressive stress which presided during emplacement. Field relationships show the long axis of the Carna Pluton lies parallel to mulitple NNW-SSE shear zones. These are represented on a regional scale by the Clifden-Mace Fault which cross cuts the core of this intrusion. AMS and petrographic data show concentric emplacement fabrics were tectonically overprinted as magma cooled from the magmatic state due to this faulting. It is proposed that the Clifden-Mace Fault system was active during ascent and emplacement of the magma and that pluton inflation only terminated as this controlling structure went into compression due to the onset of regional transtension. U-Pb zircon laser ablation inductively coupled mass spectrometry (LA-ICP-MS) data has been compiled from four sample sites. New geochronological data from the Roundstone Pluton (RD1 = ± 3.2Ma) represent the oldest age determination obtained from any member of the GGC and demonstrates that this pluton predates the Carna Pluton by ~10Ma and probably intruded synchronously with the Omey Pluton (~422.5 ± 1.7Ma). Chronological data from the Carna Pluton (CN2 = 412.9 ± 2.5Ma; CN3 = 409.8 ± 7.2Ma; CN4 = 409.6 ± 3.6Ma) represent the first precise magma crystallisation age for this intrusion. This work shows this pluton is 10Ma older than the Kilkieran Pluton and that the supply of magma into the Carna Pluton had terminated by ~409Ma. Chronological, magnetic and field data have been utilised to evaluate the kinematic evolution of the Caledonides of western Ireland throughout the construction of the GGC. It is proposed that the GGC was constructed during four distinct episodes. The style of emplacement and the conduits used for magma transport to the site of emplacement was dependent on the orientation of local structures relative to the regional ambiant stress field. This philosophy is used to critically evaluate and progress existing hypotheses on the transition from regional transpression to regional transtension at the end of the Caledonian Orogeny.

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BRCA1 has been implicated in numerous DNA repair pathways that maintain genome integrity, however the function responsible for its tumor suppressor activity in breast cancer remains obscure. To identify the most highly conserved of the many BRCA1 functions, we screened the evolutionarily distant eukaryote Saccharomyces cerevisiae for mutants that suppressed the G1 checkpoint arrest and lethality induced following heterologous BRCA1 expression. A genome-wide screen in the diploid deletion collection combined with a screen of ionizing radiation sensitive gene deletions identified mutants that permit growth in the presence of BRCA1. These genes delineate a metabolic mRNA pathway that temporally links transcription elongation (SPT4, SPT5, CTK1, DEF1) to nucleopore-mediated mRNA export (ASM4, MLP1, MLP2, NUP2, NUP53, NUP120, NUP133, NUP170, NUP188, POM34) and cytoplasmic mRNA decay at P-bodies (CCR4, DHH1). Strikingly, BRCA1 interacted with the phosphorylated RNA polymerase II (RNAPII) carboxy terminal domain (P-CTD), phosphorylated in the pattern specified by the CTDK-I kinase, to induce DEF1-dependent cleavage and accumulation of a RNAPII fragment containing the P-CTD. Significantly, breast cancer associated BRCT domain defects in BRCA1 that suppressed P-CTD cleavage and lethality in yeast also suppressed the physical interaction of BRCA1 with human SPT5 in breast epithelial cells, thus confirming SPT5 as a relevant target of BRCA1 interaction. Furthermore, enhanced P-CTD cleavage was observed in both yeast and human breast cells following UV-irradiation indicating a conserved eukaryotic damage response. Moreover, P-CTD cleavage in breast epithelial cells was BRCA1-dependent since damage-induced P-CTD cleavage was only observed in the mutant BRCA1 cell line HCC1937 following ectopic expression of wild type BRCA1. Finally, BRCA1, SPT5 and hyperphosphorylated RPB1 form a complex that was rapidly degraded following MMS treatment in wild type but not BRCA1 mutant breast cells. These results extend the mechanistic links between BRCA1 and transcriptional consequences in response to DNA damage and suggest an important role for RNAPII P-CTD cleavage in BRCA1-mediated cancer suppression.

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BACKGROUND: Scythe/BAT3 is a member of the BAG protein family whose role in apoptosis has been extensively studied. However, since the developmental defects observed in Bat3-null mouse embryos cannot be explained solely by defects in apoptosis, we investigated whether BAT3 is also involved in cell-cycle progression. METHODS/PRINCIPAL FINDINGS: Using a stable-inducible Bat3-knockdown cellular system, we demonstrated that reduced BAT3 protein level causes a delay in both G1/S transition and G2/M progression. Concurrent with these changes in cell-cycle progression, we observed a reduction in the turnover and phosphorylation of the CDK inhibitor p21, which is best known as an inhibitor of DNA replication; however, phosphorylated p21 has also been shown to promote G2/M progression. Our findings indicate that in Bat3-knockdown cells, p21 continues to be synthesized during cell-cycle phases that do not normally require p21, resulting in p21 protein accumulation and a subsequent delay in cell-cycle progression. Finally, we showed that BAT3 co-localizes with p21 during the cell cycle and is required for the translocation of p21 from the cytoplasm to the nucleus during the G1/S transition and G2/M progression. CONCLUSION: Our study reveals a novel, non-apoptotic role for BAT3 in cell-cycle regulation. By maintaining a low p21 protein level during the G1/S transition, BAT3 counteracts the inhibitory effect of p21 on DNA replication and thus enables the cells to progress from G1 to S phase. Conversely, during G2/M progression, BAT3 facilitates p21 phosphorylation by cyclin A/Cdk2, an event required for G2/M progression. BAT3 modulates these pro- and anti-proliferative roles of p21 at least in part by regulating cyclin A abundance, as well as p21 translocation between the cytoplasm and the nucleus to ensure that it functions in the appropriate intracellular compartment during each phase of the cell cycle.

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Proapoptotic Bcl-2 family members, such as Bax, promote release of cytochrome c from mitochondria, leading to caspase activation and cell death. It was previously reported that modulator of apoptosis protein 1 (MOAP-1), an enhancer of Bax activation induced by DNA damage, is stabilized by Trim39, a protein of unknown function. In this paper, we show that MOAP-1 is a novel substrate of the anaphase-promoting complex (APC/C(Cdh1)) ubiquitin ligase. The influence of Trim39 on MOAP-1 levels stems from the ability of Trim39 (a RING domain E3 ligase) to directly inhibit APC/C(Cdh1)-mediated protein ubiquitylation. Accordingly, small interfering ribonucleic acid-mediated knockdown of Cdh1 stabilized MOAP-1, thereby enhancing etoposide-induced Bax activation and apoptosis. These data identify Trim39 as a novel APC/C regulator and provide an unexpected link between the APC/C and apoptotic regulation via MOAP-1.

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Tripartite motif 39 (Trim39) is a RING domain-containing E3 ubiquitin ligase able to inhibit the anaphase-promoting complex (APC/C) directly. Through analysis of Trim39 function in p53-positive and p53-negative cells, we have found, surprisingly, that p53-positive cells lacking Trim39 could not traverse the G1/S transition. This effect did not result from disinhibition of the APC/C. Moreover, although Trim39 loss inhibited etoposide-induced apoptosis in p53-negative cells, apoptosis was enhanced by Trim39 knockdown in p53-positive cells. Furthermore, we show here that the Trim39 can directly bind and ubiquitylate p53 in vitro and in vivo, leading to p53 degradation. Depletion of Trim39 significantly increased p53 protein levels and cell growth retardation in multiple cell lines. We found that the relative importance of Trim39 and the well-characterized p53-directed E3 ligase, murine double minute 2 (MDM2), varied between cell types. In cells that were relatively insensitive to the MDM2 inhibitor, nutlin-3a, apoptosis could be markedly enhanced by siRNA directed against Trim39. As such, Trim39 may serve as a potential therapeutic target in tumors with WT p53 when MDM2 inhibition is insufficient to elevate p53 levels and apoptosis.