Participation of DNA repair in the response to 5-fluorouracil

The anti-metabolite 5-fluorouracil (5-FU) is employed clinically to manage solid tumors including colorectal and breast cancer. Intracellular metabolites of 5-FU can exert cytotoxic effects via inhibition of thymidylate synthetase, or through incorporation into RNA and DNA, events that ultimately activate apoptosis. In this review, we cover the current data implicating DNA repair processes in cellular responsiveness to 5-FU treatment. Evidence points to roles for base excision repair (BER) and mismatch repair (MMR). However, mechanistic details remain unexplained, and other pathways have not been exhaustively interrogated. Homologous recombination is of particular interest, because it resolves unrepaired DNA intermediates not properly dealt with by BER or MMR. Furthermore, crosstalk among DNA repair pathways and S-phase checkpoint signaling has not been examined. Ongoing efforts aim to design approaches and reagents that (i) approximate repair capacity and (ii) mediate strategic regulation of DNA repair in order to improve the efficacy of current anticancer treatments. Received 08 September 2008; received after revision 25 September 2008; accepted 03 October 2008     

两种工业用Al_2O_3吸附剂的结构和性质的研究

对用于净化对苯二甲酸生产中循环氮气的氧化铝吸附剂G(德国制)和W(国产),进行了结构和性质的研究。根据经不同温度熔烧的样品的X光衍射图,确定G的主要物相为η-Al_2O_3,混有少量薄水铝石;W的主要物相为薄水铝石,混有少量拟薄水铝石。这一结论又通过再水合后样品的X光衍射图得到肯定。这两种氧化铝吸附剂在孔结构,表面性质和吸附性能上都显示出明显的差别。故在应用时不宜轻易互换替代。     

Intrinsic motions along an enzymatic reaction trajectory

The mechanisms by which enzymes achieve extraordinary rate acceleration and specificity have long been of key interest in biochemistry. It is generally recognized that substrate binding coupled to conformational changes of the substrate-enzyme complex aligns the reactive groups in an optimal environment for efficient chemistry. Although chemical mechanisms have been elucidated for many enzymes, the question of how enzymes achieve the catalytically competent state has only recently become approachable by experiment and computation. Here we show crystallographic evidence for conformational substates along the trajectory towards the catalytically competent 'closed' state in the ligand-free form of the enzyme adenylate kinase. Molecular dynamics simulations indicate that these partially closed conformations are sampled in nanoseconds, whereas nuclear magnetic resonance and single-molecule fluorescence resonance energy transfer reveal rare sampling of a fully closed conformation occurring on the microsecond-to-millisecond timescale. Thus, the larger-scale motions in substrate-free adenylate kinase are not random, but preferentially follow the pathways that create the configuration capable of proficient chemistry. Such preferred directionality, encoded in the fold, may contribute to catalysis in many enzymes.     

A topographically forced asymmetry in the martian circulation and climate

Large seasonal and hemispheric asymmetries in the martian climate system are generally ascribed to variations in solar heating associated with orbital eccentricity. As the orbital elements slowly change (over a period of >104 years), characteristics of the climate such as dustiness and the vigour of atmospheric circulation are thought to vary, as should asymmetries in the climate (for example, the deposition of water ice at the northern versus the southern pole). Such orbitally driven climate change might be responsible for the observed layering in Mars' polar deposits by modulating deposition of dust and water ice. Most current theories assume that climate asymmetries completely reverse as the angular distance between equinox and perihelion changes by 180 degrees. Here we describe a major climate mechanism that will not precess in this way. We show that Mars' global north-south elevation difference forces a dominant southern summer Hadley circulation that is independent of perihelion timing. The Hadley circulation, a tropical overturning cell responsible for trade winds, largely controls interhemispheric transport of water and the bulk dustiness of the atmosphere. The topography therefore imprints a strong handedness on climate, with water ice and the active formation of polar layered deposits more likely in the north.     

Flying in a flock comes at a cost in pigeons

Flying birds often form flocks, with social, navigational and anti-predator implications. Further, flying in a flock can result in aerodynamic benefits, thus reducing power requirements, as demonstrated by a reduction in heart rate and wingbeat frequency in pelicans flying in a V-formation. But how general is an aerodynamic power reduction due to group-flight? V-formation flocks are limited to moderately steady flight in relatively large birds, and may represent a special case. What are the aerodynamic consequences of flying in the more usual 'cluster' flock? Here we use data from innovative back-mounted Global Positioning System (GPS) and 6-degrees-of-freedom inertial sensors to show that pigeons (1) maintain powered, banked turns like aircraft, imposing dorsal accelerations of up to 2g, effectively doubling body weight and quadrupling induced power requirements; (2) increase flap frequency with increases in all conventional aerodynamic power requirements; and (3) increase flap frequency when flying near, particularly behind, other birds. Therefore, unlike V-formation pelicans, pigeons do not gain an aerodynamic advantage from flying in a flock. Indeed, the increased flap frequency, whether due to direct aerodynamic interactions or requirements for increased stability or control, suggests a considerable energetic cost to flight in a tight cluster flock.     

Base excision repair and design of small molecule inhibitors of human DNA polymerase β

Base excision repair (BER) can protect a cell after endogenous or exogenous genotoxic stress, and a deficiency in BER can render a cell hypersensitive to stress-induced apoptotic and necrotic cell death, mutagenesis, and chromosomal rearrangements. However, understanding of the mammalian BER system is not yet complete as it is extraordinarily complex and has many back-up processes that complement a deficiency in any one step. Due of this lack of information, we are unable to make accurate predictions on therapeutic approaches targeting BER. A deeper understanding of BER will eventually allow us to conduct more meaningful clinical interventions. In this review, we will cover historical and recent information on mammalian BER and DNA polymerase β and discuss approaches toward development and use of small molecule inhibitors to manipulate BER. With apologies to others, we will emphasize results obtained in our laboratory and those of our collaborators.     

鱼类营养学研究的普遍问题及建议

在确定鱼类对潜在营养源的利用或营养物需求的研究中,常存在一些问题.本文将讨论这些问题,并提出这类实验的一般步骤. 1、饲料 商用饲料需注明饲料及厂家的全称.实验饲料需列出所有原料的标准名称及国际饲料号.精饲料及添加剂需列出原料供应商及地址.基础饲料应能提供实验鱼所需的全部营养.除了所测成分外,饲料的组成应是恒定的.最好能根据消化率数据确定饲料配方.饲料制作的步骤应描述清楚.饲料使用前应冷冻保存.饲料应采用国际上接受的方法进行化学分析. 2、实验条件 应描述饲养单元的大小、位置、水源及水处理方法、水温、流速、溶氧、水质参数、光照条件、实验鱼的详细情况及其他实验条件.养鱼系统包括流水、循环过滤水及静水系统.静水系统不能用于营养需求实验,循环水系统亦不宜用于维生素及无机盐需求量实验. 3、实验设计 每一饲养单元是一个重复,单元内的个体不能看作重复样.每一处理至少需要3个重复.实验鱼需经过一段时间暂养才能用于实验.实验周期至少8周,微营养物需求实验需要12-20周.实验期增重率应在200%-500%以上.饲养单元应随机分配于各个处理.投喂方式可以是饱食或限食.若采用限食投喂,投喂水平应接近饱食. 4、数据评价 如果对照鱼生长率较预期的低,或死亡率较高,或实验结果与预期的不同,则有必要重复实验.需求量实验应得到剂量反应曲线,否则须重新设计实验.建议采用拐点法或回归法确定最低需求量.     

The status of Wnt signalling regulates neural and epidermal fates in the chick embryo

The acquisition of neural fate by embryonic ectodermal cells is a fundamental step in the formation of the vertebrate nervous system. Neural induction seems to involve signalling by fibroblast growth factors (FGFs) and attenuation of the activity of bone morphogenetic protein (BMP). But FGFs, either alone or in combination with BMP antagonists, are not sufficient to induce neural fate in prospective epidermal ectoderm of amniote embryos. These findings suggest that additional signals are involved in the specification of neural fate. Here we show that the state of Wnt signalling is a critical determinant of neural and epidermal fates in the chick embryo. Continual Wnt signalling blocks the response of epiblast cells to FGF signals, permitting the expression and signalling of BMP to direct an epidermal fate. Conversely, a lack of exposure of epiblast cells to Wnt signals permits FGFs to induce a neural fate.     

Rapid cloning of high-affinity human monoclonal antibodies against influenza virus

Pre-existing neutralizing antibody provides the first line of defence against pathogens in general. For influenza virus, annual vaccinations are given to maintain protective levels of antibody against the currently circulating strains. Here we report that after booster vaccination there was a rapid and robust influenza-specific IgG+ antibody-secreting plasma cell (ASC) response that peaked at approximately day 7 and accounted for up to 6% of peripheral blood B cells. These ASCs could be distinguished from influenza-specific IgG+ memory B cells that peaked 14-21 days after vaccination and averaged 1% of all B cells. Importantly, as much as 80% of ASCs purified at the peak of the response were influenza specific. This ASC response was characterized by a highly restricted B-cell receptor (BCR) repertoire that in some donors was dominated by only a few B-cell clones. This pauci-clonal response, however, showed extensive intraclonal diversification from accumulated somatic mutations. We used the immunoglobulin variable regions isolated from sorted single ASCs to produce over 50 human monoclonal antibodies (mAbs) that bound to the three influenza vaccine strains with high affinity. This strategy demonstrates that we can generate multiple high-affinity mAbs from humans within a month after vaccination. The panel of influenza-virus-specific human mAbs allowed us to address the issue of original antigenic sin (OAS): the phenomenon where the induced antibody shows higher affinity to a previously encountered influenza virus strain compared with the virus strain present in the vaccine. However, we found that most of the influenza-virus-specific mAbs showed the highest affinity for the current vaccine strain. Thus, OAS does not seem to be a common occurrence in normal, healthy adults receiving influenza vaccination.