基于核酸侵入反应的生物分子检测技术的研究进展

核酸侵入反应是由5＇核酸内切酶或flap内切酶催化的,能够识别切割核酸片段形成的特异性结构的一类反应。近年来发展了很多基于该反应的生物大分子检测技术,能够对DNA、RNA、miRNA及蛋白质进行高灵敏、高特异性的测定。这些技术大都无需扩增待测靶标,极大地降低了扩增产物交叉污染的风险,在临床检测中具有很大的应用前景。本文对这些检测技术的原理及应用作简要综述。     

基于核酸侵入反应的生物分子检测技术的研究进展

核酸侵入反应是由5’核酸内切酶或flap内切酶催化的,能够识别切割核酸片段形成的特异性结构的一类反应。近年来发展了很多基于该反应的生物大分子检测技术,能够对DNA、RNA、miRNA及蛋白质进行高灵敏、高特异性的测定。这些技术大都无需扩增待测靶标,极大地降低了扩增产物交叉污染的风险,在临床检测中具有很大的应用前景。本文对这些检测技术的原理及应用作简要综述。     

“It was the photograph of the little boy”: reflections on the Syrian Vulnerable Persons Resettlement Programme in the UK

This article examines the “Syrian Vulnerable Persons Resettlement Programme” (SVPRP) as a specific British response to the “European refugee crisis”. Based on an analysis of media reporting (2014–17) and empirical evidence from agencies and volunteers tasked with implementing the programme, this essay reveals the ethical and political ambiguities at its heart. By focusing on the notion of “vulnerability” I argue that the humanitarian configuration of a refugee worthy of care is implicated in two significant practices: exceptionalising a small group of Syrians as legitimate targets for compassion and constructing compassion itself as a rationed resource in a climate of anti-immigrant hostility, austerity and Brexit.     

Eukaryotic lagging strand DNA replication employs a multi-pathway mechanism that protects genome integrity

In eukaryotic nuclear DNA replication, one strand of DNA is synthesized continuously, but the other is made as Okazaki fragments that are later joined. Discontinuous synthesis is inherently more complex, and fragmented intermediates create risks for disruptions of genome integrity. Genetic analyses and biochemical reconstitutions indicate that several parallel pathways evolved to ensure that the fragments are made and joined with integrity. An RNA primer is removed from each fragment before joining by a process involving polymerase-dependent displacement into a single-stranded flap. Evidence in vitro suggests that, with most fragments, short flaps are displaced and efficiently cleaved. Some flaps can become long, but these are also removed to allow joining. Rarely, a flap can form structure, necessitating displacement of the entire fragment. There is now evidence that post-translational protein modification regulates the flow through the pathways to favor protection of genomic information in regions of actively transcribed chromatin.     

The Messinian event: What happened to the peri-Mediterranean mammalian communities and local climate?

What truly happened in the terrestrial ecosystems in response to the famous Messinian salinity crisis is still the matter of extensive discussions. Did mammals record any faunal and/or a climatic or environmental fluctuation is a question that still remains open. Our objective is to investigate mammalian faunas before, during and after the crisis within different terrestrial basins surrounding the Mediterranean Sea. We therefore apply here two methods, the cenogram method and a transfer function based on murine (rodents) species richness to better understand if our proxies can record any qualitative or quantitative climatic or environmental change. The results indicate that mammal faunas do not record any particular shift in climate or environment at the scale of the whole peri-Mediterranean area. The trend is different at the regional scale of terrestrial basins as temperatures increase (Calatayud-Daroca-Teruel Basin) or decrease (Languedoc-Roussillon region) punctually occurs just after the crisis; stable conditions in different areas (e.g. Greece) contradict these trends. Biases in the fossil record or in the methodologies could produce such discrepancies.     

Is the keystone role of figs maintained across a gradient of increasing human disturbance?

Fig trees provide important resources to tropical animals, but is their keystone role affected by human disturbance? In Sri Lanka, we found that more bird species were found in close association with fruiting figs in urban areas, suggesting their role is accentuated there, and justifying their planting.     

Opportunism and Diversification: Entrepreneurship and Livelihood Strategies in Uncertain Times

As economic crisis deepens across Europe people are forced to find innovative strategies to accommodate circumstances of chronic uncertainty. Even with a second multi-billion euro bailout package secured for Greece, the prospects of a sustainable recovery in the near future look bleak. However, crisis has also created dynamic spaces for entrepreneurial opportunism and diversification resulting in social mobility, relocation, shifts in livelihood strategy and a burgeoning informal economy. Although economic systems are currently undergoing radical reassessment, social demands such as competitive consumption remain. Opportunities for investment in renewable energy programmes, especially photovoltaics, are also pervasive. By considering cases of business opportunism and livelihood diversification in relation to Max Weber's concept of wertrational and notions of uncertainty, this article brings new perspectives to strategies of negotiating the worst economic crisis in living memory.     

Flap Endonuclease Activity of Gene 6 Exonuclease of Bacteriophage T7

Flap endonucleases remove flap structures generated during DNA replication. Gene 6 protein of bacteriophage T7 is a 5′–3′-exonuclease specific for dsDNA. Here we show that gene 6 protein also possesses a structure-specific endonuclease activity similar to known flap endonucleases. The flap endonuclease activity is less active relative to its exonuclease activity. The major cleavage by the endonuclease activity occurs at a position one nucleotide into the duplex region adjacent to a dsDNA-ssDNA junction. The efficiency of cleavage of the flap decreases with increasing length of the 5′-overhang. A 3′-single-stranded tail arising from the same end of the duplex as the 5′-tail inhibits gene 6 protein flap endonuclease activity. The released flap is not degraded further, but the exonuclease activity then proceeds to hydrolyze the 5′-terminal strand of the duplex. T7 gene 2.5 single-stranded DNA-binding protein stimulates the exonuclease and also the endonuclease activity. This stimulation is attributed to a specific interaction between the two proteins because Escherichia coli single-stranded DNA binding protein does not produce this stimulatory effect. The ability of gene 6 protein to remove 5′-terminal overhangs as well as to remove nucleotides from the 5′-termini enables it to effectively process the 5′-termini of Okazaki fragments before they are ligated.     

LY294002对CML急变期细胞中Wnt／β-catenin信号通路的影响及其效应

β-catening在慢性粒细胞白血病急变过程中发挥着重要作用,而其受BCR／ABLTL其下游信号通路调控的具体分子机制尚未完全阐明。该研究旨在探讨PI3K-AK聪号通路对慢粒急变期细胞的影响及其对Wnt／β-catenin信号通路的调控作用。采用P13K-AKT信号通路的靶向抑制剂LY294002作用于慢粒急变期K562细胞,MTT法检测其对细胞增殖的影响,甲基纤维素克隆形成实验检测细胞的克隆形成能力,Western blot检测pAKT（Thr308）的表达变化,RT-PCR和Western blot分别检测β-catenin及其下游靶基因c—myc、cyclinD1的mRNA和蛋白表达情况。结果显示,10,20,40μmol/L的LY294002作用细胞24h后,抑制了K562细胞的增殖以及克隆形成能力。该效应呈浓度依赖的方式。3种浓度的LY294002处理细胞后,PI3K—AKT信号通路明显被抑制,pAKT（Thr308）的蛋白表达明显减少;β-catenin的mRNA表达无明显改变,但其蛋白水平依次减少;β-catenin的下游靶基因c-myc、cyclin D1的mRNA和蛋白水平均明显降低。综上所述,抑制PI3K-AKT信号通路可抑制白血病K562细胞的增殖和克隆形成能力,其机制可能与抑制Wnt／β-catenin信号通路相关。     

Homogenization of freshwater fish faunas after the elimination of a natural barrier by a dam in Neotropics

Aim We quantify biotic homogenization of fish fauna caused by the elimination of a natural barrier between two freshwater ecoregions. We also evaluated fish introductions by different mechanisms such as aquaculture, angling and the aquarium trade in the homogenization of fish assemblages. The relative importance of native extinctions in the homogenization process was assessed by simulating the exclusion of threatened species in the data set. Location Paraná River, south‐eastern South America. Methods A fish species list of the Parana River Basin was organized in a subset of species distributions, according to pre‐ and post‐introductions caused by the elimination of the natural barrier and by other mechanisms. Biotic homogenization was verified by the use of Jaccard’s and Bray–Curtis’s coefficients, Whittaker’s beta diversity index, non‐metric multidimensional scaling analysis (NMDS) and nonparametric tests. Results For all subsets of species distributions, we observed an increase in the number of non‐native species in common related to the introductions. Between 40 and 52% of the species currently present in the Upper Paraná Basin dispersed upstream from the Lower Paraná after the construction of Itaipu Dam, including at least 1 class, 2 orders, 4 families and 16 genera of fish. Jaccard’s coefficient between the Upper and Lower Parana River increased by 6–7.5% only considering the Itaipu Dam influence and 10.5% considering all mechanisms of fish introductions. More than 50% of the increase in similarity was caused by the elimination of the barrier. Our results indicated functional homogenization related to large‐bodied Siluriformes (catfish). Main conclusions Itaipu Lake flooded a natural barrier and allowed hydrologic connectivity between the Upper and Lower Paraná River, and many fishes of the lower part of the river were able to colonize the upper stretches. The homogenization of the two assemblages between these adjacent aquatic regions was an unpredicted result of hydropower implementation. Introductions by dam can also shift longitudinal and latitudinal body size patterns (i.e. Bergmann's rule).