GST pull-down筛选冠突曲霉MAT的互作蛋白*

目的: 冠突曲霉(Aspergillus cristatus)是一种同宗结合菌,它的产孢受渗透压调控,与构巢曲霉的光调控产孢机制存在较大差异。冠突曲霉的有性生殖主要受MAT1-1-1和MAT1-2-1调控,但MAT基因对该菌有性生殖的调控机制仍不清楚。期望筛选得到冠突曲霉MAT的互作蛋白,为深入研究冠突曲霉有性产孢机制奠定基础。方法: 利用GST pull-down联合液相色谱-串联质谱(LC-MS/MS)技术筛选可能与冠突曲霉MAT1-1-1和MAT1-2-1互作的蛋白,结合ProteinPilot和冠突曲霉基因组注释结果进行互作蛋白的注释及GO分析,其中互作蛋白SI65_00917和SI65_03348利用RT-qPCR探索它们与有性发育的联系,并利用酵母双杂交技术初步验证它们与MAT蛋白的互作关系。结果: 成功构建了GST-MAT1-1-1、GST-MAT1-2-1表达载体,诱导表达纯化出目的诱饵蛋白,分别利用诱饵蛋白捕获冠突曲霉总蛋白中的互作蛋白,经分析、筛选共鉴定出与MAT1-1-1互作的蛋白56个,与MAT1-2-1互作的蛋白413个。GO分析表明,这些蛋白参与翻译调控、代谢过程、蛋白质转运及蛋白结合等生物学过程,具有核苷酸结合活性、催化活性、蛋白结合活性;RT-qPCR结果表明互作蛋白SI65_00917可能与有性发育相关。酵母双杂交结果表明,SI65_00917蛋白具有自激活作用,可能是转录因子;SI65_03348蛋白与MAT1-1-1、MAT1-2-1在酵母中均有互作。结论: MAT通过与其他蛋白直接或间接的相互作用调控其有性发育过程。     

樟子松固沙林林水关系研究进展及对营林实践的指导

中国有着世界上最大面积的人工林, 如何维持人工林的可持续性已成为气候变化背景下需要面对的重大挑战。樟子松(Pinus sylvestris var. mongolica)以其抗旱、抗寒、耐贫瘠等优良特性成为中国北方生态治理中最主要的常绿针叶树种之一, 近70年来发挥了巨大的防风固沙与生态固碳功能。然而, 随着林分的生长与气候变化, 樟子松人工固沙林正经历着越来越严峻的环境胁迫, 部分地区出现了林分“早衰”或死亡的现象, 引起了人们对樟子松固沙林适应与应对气候变化能力的担忧。该文在回溯樟子松基本生物学特征与引种推广历史, 系统总结近年来樟子松林林水关系研究新成果的基础上, 全面分析了樟子松固沙林林水关系存在的主要矛盾, 并提出了基于林水关系相协调的林分经营措施的调整: 由倡导防护功能为主的单一目标向包含林分稳定性、生态固碳功能、可持续发展等多目标平衡方向调整; 由以沙地森林景观培育为主向以良好土壤生境培育为主的方向调整; 由倡导天然更新为主向以人工造林与天然更新相结合的世代更新方向调整。在立足于北方沙地脆弱生境与气候变化客观现实的基础背景下, 应坚持樟子松在固沙林生态系统演化过程中的先锋种与建群种地位, 基于“以水定绿”原则, 采取“隔行带伐+再造林”等方式开展林分密度动态调控, 促进林分向异龄林结构演化, 促进樟子松固沙林生态服务的优质化和生态固碳功能的最大化。     

中国真猛犸象和披毛犀化石14C年代研究新进展

真猛犸象（Mammuthus primigenius）和披毛犀（Coelodonta antiquitatis）是北半球高纬度地区晚更新世动物群的主要成员,其消亡的年代和原因一直是国际学术界关注的热点科学问题。本文对黑龙江青冈县英贤村最新出土的5个真猛犸象和5个披毛犀化石进行了AMS¹⁴C年代测定,结果均大于4万年,部分化石可能已经超出了目前¹⁴C的测定范围。通过整理并对比已公开发表的中国境内两种动物化石的¹⁴C年代学数据,本文认为早期常规¹⁴C测年方法所获得的年代值需要重新考虑其准确性。埋藏地层与最新的AMS¹⁴C测年数据显示,我国真猛犸象化石年代主要集中于MIS3阶段;披毛犀在我国消亡的时间很可能晚于真猛犸象,至少延续到末次冰消期。中国猛犸象-披毛犀动物群化石仍然需要开展更多的年代学研究。     

A Biomechanical Model of Tumor-Induced Intracranial Pressure and Edema in Brain Tissue

Brain tumor growth and tumor-induced edema result in increased intracranial pressure (ICP), which, in turn, is responsible for conditions as benign as headaches and vomiting or as severe as seizures, neurological damage, or even death. Therefore, it has been hypothesized that tracking ICP dynamics may offer improved prognostic potential in terms of early detection of brain cancer and better delimitation of the tumor boundary. However, translating such theory into clinical practice remains a challenge, in part because of an incomplete understanding of how ICP correlates with tumor grade. Here, we propose a multiphase mixture model that describes the biomechanical response of healthy brain tissue—in terms of changes in ICP and edema—to a growing tumor. The model captures ICP dynamics within the diseased brain and accounts for the ability/inability of healthy tissue to compensate for this pressure. We propose parameter regimes that distinguish brain tumors by grade, thereby providing critical insight into how ICP dynamics vary by severity of disease. In particular, we offer an explanation for clinically observed phenomena, such as a lack of symptoms in low-grade glioma patients versus a rapid onset of symptoms in those with malignant tumors. Our model also takes into account the effects tumor-derived proteases may have on ICP levels and the extent of tumor invasion. This work represents an important first step toward understanding the mechanisms that underlie the onset of edema and ICP in cancer-afflicted brains. Continued modeling effort in this direction has the potential to make an impact in the field of brain cancer diagnostics.     

大肠杆菌代谢工程改造合成L-高丝氨酸及其衍生物研究进展

l-高丝氨酸及其衍生物(O-琥珀酰-l-高丝氨酸和O-乙酰-l-高丝氨酸)是生物合成l-甲硫氨酸的前体,同时也是合成多种C4化合物(异丁醇、g-丁内酯、1,4-丁二醇、2,4-二羟基丁酸等)和l-草铵膦等的平台化合物。因此,发酵法生产l-高丝氨酸及其衍生物成为近年内研究的热点。然而,利用生物法合成l-高丝氨酸及其衍生物仍存在一些不足之处,如发酵产量不高或糖酸转化率过低等。此外,对l-高丝氨酸及其衍生物合成的总体代谢和调控机制鲜有报道。本文综述了大肠杆菌代谢工程改造合成l-高丝氨酸及其衍生物O-琥珀酰-l-高丝氨酸和O-乙酰-l-高丝氨酸的研究进展,从底物摄取、关键节点碳流分配改造、辅酶NADPH的循环供应以及目标产物的外运输出等方面,系统分析了大肠杆菌全发酵法生产l-高丝氨酸及其衍生物的代谢途径及改造策略,为其后续代谢改造及生物法生产提供一定的研究思路。     

Structure of cytochrome c oxidase: a comparison of the bacterial and mitochondrial enzymes

It has been almost 5 years since the first structures of cytochrome c oxidase, from Paracoccus denitrificans and bovine heart mitochondria, were revealed. Since then many different proton pumping mechanisms have been proposed for the enzyme; however, no definitive conclusion has been achieved. In this article, we revisit the original structures of bacterial and mitochondrial oxidases and try to clarify similarities as well as differences between the two structures.     

An insensitive acetylcholinesterase confers resistance to methomyl in the beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae)

Two forms of acetylcholinesterase were identified in field populations of the beet armyworm, Spodoptera exigua (Hübner), collected from cotton in San Joaquin Valley, CA. Strains (BESS and BKRR) homogeneous for each variant were isolated and their relative susceptibilities to methomyl, chlorpyrifos, and chlorpyrifos-oxon assessed by topical application bioassay. In comparisons with a laboratory susceptible strain (DOW), BKRR and BESS expressed 68-fold and sevenfold resistance, respectively, to the carbamate methomyl. Neither strain was cross-resistant to chlorpyrifos or its oxygen analog (chlorpyrifos-oxon). In biochemical studies, the BKRR AChE enzyme was approximately 30-fold and sixfold more insensitive to methomyl and chlorpyrifos-oxon, respectively, compared with the DOW enzyme. The correlation between the toxicological and biochemical studies provides strong evidence that target-site insensitivity is an important mechanism of resistance to methomyl. The lack of significant cross-resistance to chlorpyrifos suggests that the insensitive AChE in these field populations was selected by methomyl alone and not by the organophosphate.     

Il-17 mobilizes peripheral blood stem cells with short- and long-term repopulating ability in mice

Autologous and allogeneic bone marrow transplantations have evolved as important cancer therapy modalities. For both indications, peripheral blood has been shown to have distinct advantages over bone marrow as the stem cell source. Cytokine combinations for mobilization have enhanced stem cell yield and accelerated engraftment. However, novel mobilizing agents and strategies are needed to further improve clinical outcomes. Within the donor graft, the dynamic equilibrium between T cells and stem cells critically influences engraftment and transplantation results. IL-17 is a cytokine produced almost exclusively from activated T cells. IL-17 was expressed in vivo with adenovirus technology. Here, proof-of-principle studies demonstrate that IL-17 effectively mobilizes hemopoietic precursor cells (CFU-granulocyte-erythrocyte-macrophage-monocyte, CFU-high proliferative potential) and primitive hemopoietic stem cells (Lin(-/low)c-kit(+)Sca1(+)). Moreover, mouse IL-17 adenovirus-mobilized peripheral blood stem cells rescued lethally irradiated mice. Bone marrow was found to be 45-75% of donor origin at 1 year. In secondary recipients, donor-derived bone marrow cells ranged from 45 to 95%. These data show that IL-17 mobilizes stem cells in mice with short- and long-term reconstituting capacity. Additional comparative studies are needed as well as studies in tumor models to refine distinct potential clinical applications for IL-17-mobilized peripheral blood stem cells.