Ubiquitinated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis

Ubiquitin-positive, tau- and alpha-synuclein-negative inclusions are hallmarks of frontotemporal lobar degeneration with ubiquitin-positive inclusions and amyotrophic lateral sclerosis. Although the identity of the ubiquitinated protein specific to either disorder was unknown, we showed that TDP-43 is the major disease protein in both disorders. Pathologic TDP-43 was hyper-phosphorylated, ubiquitinated, and cleaved to generate C-terminal fragments and was recovered only from affected central nervous system regions, including hippocampus, neocortex, and spinal cord. TDP-43 represents the common pathologic substrate linking these neurodegenerative disorders.     

农业土地系统研究及其关键科学问题

如何科学合理利用土地是实现人类可持续发展的关键。随着人类对土地利用问题愈发关注,在“土地利用/土地覆盖变化”、“全球土地计划”等国际科学研究计划的推动下,“土地系统科学”的学科体系逐步形成,农业土地系统研究成为土地系统科学的热点方向之一。本文以全球土地计划与土地系统科学为指引,旨在明晰“农业土地系统”的概念,系统梳理农业土地系统研究的技术方法、内容对象以及关键科学问题,为进一步丰富和完善土地系统科学学科体系,推动全球变化、粮食安全及农业可持续研究提供参考。研究认为：第一,多维度格局探测与分析是农业土地系统研究的重要基础：农业土地系统不仅关注耕地与其他土地利用类型之间的相互转换特征、规律和过程,而且更为关注耕地内部多熟种植制度、农作物空间格局、利用集约度、综合生产能力等结构和功能的多维变化,因此,需要依靠多学科/数据的交汇、融合等手段来揭示农业土地系统的复杂特征;第二,多模型耦合的过程与机制解析是农业土地系统研究的核心内容：在明晰农业土地系统时空格局特征的基础上,通过建立土地系统格局与其影响因素之间的关系,并将这种关系在时间维度进行扩展,进而实现农业土地系统变化过程和机制的动态表达,目前,土地变化模型的建模手段已从传统单一的地理模型或经济模型研究转向模型耦合研究,以反映农业土地系统中“人类-环境”的复杂关系;第三,多内容的综合效应评估与调控是农业土地系统研究的关键任务：农业土地系统与全球变化、粮食安全及农业可持续发展等问题密切关联,农业土地系统时空格局探测、过程机制解析的最终目的在于通过协调农业土地系统与农业资源、环境和生态的相互关系,考虑不同系统之间的权衡优化关系,追求土地利用的最佳社会、经济和生态综合效益,以建立人地和谐、可持续的农业土地利用模式。解决农业土地系统研究的这些关键科学问题,将有力促进自然科学和社会科学的融合,推动土地系统科学及相关研究领域的发展。     

水稻和稗草ALS活性测定及农美利选择性机理研究

乙酰乳酸合成酶（ＡＬＳ）是除草剂主要作用靶标,新型除草剂农美利（ｂｉｓｐｙｒｉｂａｃ－ｓｏｄｉｕｍ）即是通过抑制ＡＬＳ的活性使支链氨基酸的生物合成受阻而达到除草目的。作者利用ＡＬＳ脱羧产物３－羟基丁酮与肌酸及甲萘酚形成的复合物,采用比色法测定ＡＬＳ活性和农美利的抑制作用,以明确农美利对水稻和稗草的选择性机理。研究结果表明,水稻和稗草体内ＡＬＳ酶促反应复合产物的吸收峰为５２０ｎｍ;ＡＬＳ活性在不同水稻品种和不同杂划体内各不相同,不同水稻品种依次为汕优６３、秀水１１〉样湖８４〈嘉育２９３,不同杂草依次为稗草、鳢肠〉异型莎草〉千金子。体内法测定农美利对植物体内ＡＬＳ的抑制程度结果显示,水稻ＡＬＳ受抑制快于稗草,恢复也快;不同水稻品种ＡＬＳ以嘉育２９３受抑制较快,秀水１１抑制程度较大,而祥湖８４和汕优６３受抑制慢且程度     

水稻乙酰乳酸合成酶基因的克隆和功能分析

乙酰乳酸合成酶（ALS）是植物和微生物中亮氨酸、异亮氨酸和缬氨酸合成途径的一个关键酶，该基因在植物抗除草剂基因工程中具有重要的应用价值。利用生物信息学分析，自水稻中分离了ALS基因的cDNA和基因组DNA序列。序列分析表明，ALS基因没有内含子，GC碱基含量高并不对称分布。软件预测ALS基因是单拷贝基因位于第4号染色体上，Southern杂交验证了此结果。构建A岱基因的植物表达载体并转化烟草，转基因烟草通过PCR-Southern验证。EXACT-RT-PCR结果表明，水稻ALS基因在转基因烟草的叶片和根部均有转录发生。利用甲嘧磺隆进行除草剂抗性测试，和对照烟草相比转基因烟草除草剂抗性高达50mg·L^-1。     

Activity of Acetolactate Synthase from Maize (Zea mays L. ) as Influenced by Chlorsulfuron and Tribenuron-methyl

Study on relative sensitivity of maize (Zea mays L. ) Nongda108 and Nongda3138 to sulfonylurea herbicide chlorsulfuron and tribenuron-methyl using maize taproot length by sand bioassy indicated that, Nongda3138 had higher tolerance to chlorsulfuron and tribenuron-methyl than Nongda108 did. Chlorsulfuron had stronger growth inhibition to maize Nongda108 and Nongda3138 than tribenuron-methyl did. Study on target enzyme of sulfonylurea herbicide acetolactate synthase (ALS) showed that, chlorsulfuron and tribenuron-methyl inhibited ALS in vitro strongly, and non-competitively. In the same concentration of inhibitors,chlorsuifuron had stronger ALS activity inhibition than tribenuron-methyl did. Lower level of chlorsulfuron and tribenuron-methyl has no ALS activity inhibition in vivo, the ALS inhibition only occurred in the condition of high concentration of chlorsulfuron and tribenuron-methyl in vivo.     

氯磺隆和苯磺隆对玉米乙酰乳酸合成酶抑制作用的研究

 用玉米沙培主根长度法测定了玉米品种农大 3138和农大 10 8对氯磺隆和苯磺隆的相对敏感性。结果表明 ,农大 3138耐药能力比农大 10 8强 ,氯磺隆对玉米生长的影响比苯磺隆大。有关玉米离体乙酰乳酸合成酶(ALS)的研究结果表明 ,氯磺隆和苯磺隆对两种玉米离体ALS酶具有强烈的抑制作用 ,是玉米ALS酶的非竞争性抑制剂 ,氯磺隆对两种玉米ALS酶的抑制作用强于苯磺隆。低浓度的氯磺隆和苯磺隆对活体ALS酶无抑制作用 ,高浓度下才抑制活体ALS酶的活性。     

Retrograde viral delivery of IGF-1 prolongs survival in a mouse ALS model

Amyotrophic lateral sclerosis (ALS) is a progressive, lethal neuromuscular disease that is associated with the degeneration of spinal and brainstem motor neurons, leading to atrophy of limb, axial, and respiratory muscles. The cause of ALS is unknown, and there is no effective therapy. Neurotrophic factors are candidates for therapeutic evaluation in ALS. Although chronic delivery of molecules to the central nervous system has proven difficult, we recently discovered that adeno-associated virus can be retrogradely transported efficiently from muscle to motor neurons of the spinal cord. We report that insulin-like growth factor 1 prolongs life and delays disease progression, even when delivered at the time of overt disease symptoms.     

Functional role of caspase-1 and caspase-3 in an ALS transgenic mouse model

Mutations in the copper/zinc superoxide dismutase (SOD1) gene produce an animal model of familial amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disorder. To test a new therapeutic strategy for ALS, we examined the effect of caspase inhibition in transgenic mice expressing mutant human SOD1 with a substitution of glycine to alanine in position 93 (mSOD1(G93A)). Intracerebroventricular administration of zVAD-fmk, a broad caspase inhibitor, delays disease onset and mortality. Moreover, zVAD-fmk inhibits caspase-1 activity as well as caspase-1 and caspase-3 mRNA up-regulation, providing evidence for a non-cell-autonomous pathway regulating caspase expression. Caspases play an instrumental role in neurodegeneration in transgenic mSOD1(G93A) mice, which suggests that caspase inhibition may have a protective role in ALS.     

抗苯磺隆猪殃殃乙酰乳酸合成酶的突变研究

[目的]近几年,中国大部分冬小麦田猪殃殃(Galium agarine L.)用苯磺隆已无法有效控制.为明确猪殃殃对苯磺隆产生抗药性的分子机制,本试验从分子水平上开展研究,以初步明确乙酰乳酸合成酶(ALS)氨基酸序列的突变位点.[方法]通过对敏感和抗药性猪殃殃生物型ALS基因片段进行扩增、克隆和测序,比对2种生物型的ALS序列.[结果]与敏感生物型猪殃殃的ALS相比,抗药性猪殃殃生物型ALS有3个位点发生突变,其中位于高度保守区Domain B的第574位色氨酸被甘氨酸所取代.[结论]该位点的突变可能是猪殃殃对苯磺隆产生抗药性的主要原因.     

基于稀疏型机载激光雷达数据的风景林参数估测

稀疏型机载激光雷达（LiDAR）数据,由于点云密度低,难以对单木尺度的森林参数进行估测,在森林资源监测领域的潜在应用价值没有得到充分发挥。以江苏省南京市紫金山国家森林公园为研究区,以2007年机载激光雷达Optech ALTM LiDAR数据、2007年森林资源规划设计调查数据为主要信息源,在机载LiDAR数据预处理、特征参数提取的基础上,采用多元线性回归、随机森林、支持向量机3种方法,估测小班林分平均树高、平均胸径和单位蓄积量并进行对比分析,同时对森林参数进行空间制图。结果表明:1） 高度百分位数和累计高度百分位数是影响小班平均胸径、平均树高和平均蓄积量的主要特征参数;2）3个遥感估测模型精度对比分析表明,平均胸径、平均树高、单位蓄积量3个参数的估测精度,均是随机森林算法最高,支持向量机次之,多元线性回归最低;3）平均树高、平均胸径、单位蓄积量的空间分布规律一致,人为干扰严重的公园边缘地区和土层瘠薄、立地条件差的山脊较低,山南及山北中部最高。稀疏型机载激光雷达（ALS）数据在林分尺度的森林参数估计中具有较高的精度,可以用于森林资源规划设计调查小班的测树因子估测。本研究应用稀疏型机载激光雷达数据估测风景林森林参数,为稀疏型LiDAR数据在森林资源规划设计调查中的推广应用提供参考。     

Wild-type nonneuronal cells extend survival of SOD1 mutant motor neurons in ALS mice

The most common inherited [correct] form of amyotrophic lateral sclerosis (ALS), a neurodegenerative disease affecting adult motor neurons, is caused by dominant mutations in the ubiquitously expressed Cu-Zn superoxide dismutase (SOD1). In chimeric mice that are mixtures of normal and SOD1 mutant-expressing cells, toxicity to motor neurons is shown to require damage from mutant SOD1 acting within nonneuronal cells. Normal motor neurons in SOD1 mutant chimeras develop aspects of ALS pathology. Most important, nonneuronal cells that do not express mutant SOD1 delay degeneration and significantly extend survival of mutant-expressing motor neurons.     

Mutations in dynein link motor neuron degeneration to defects in retrograde transport

Degenerative disorders of motor neurons include a range of progressive fatal diseases such as amyotrophic lateral sclerosis (ALS), spinal-bulbar muscular atrophy (SBMA), and spinal muscular atrophy (SMA). Although the causative genetic alterations are known for some cases, the molecular basis of many SMA and SBMA-like syndromes and most ALS cases is unknown. Here we show that missense point mutations in the cytoplasmic dynein heavy chain result in progressive motor neuron degeneration in heterozygous mice, and in homozygotes this is accompanied by the formation of Lewy-like inclusion bodies, thus resembling key features of human pathology. These mutations exclusively perturb neuron-specific functions of dynein.     

Effect of Chlorimuron-Ethyl on Biochemical Mechanism in Tolerant Sugar Beet

Effect of chlorimuron-ethyl on biochemical mechanism in tolerant sugar beet was investigated to provide basic data on using the tolerant genotype properly. Tolerant sugar beet was used to analyze its biochemical mechanism under chlorimuron- ethyl stress with frame culture in field and water culture. Glutathione S-transferase （GST） activity of leaves in tolerant sugar beet was remarkably increased as chlorimuron-ethyl was preemergence applicated at 0.5 and 1.5 g a.i. hw1, at the same time glutathione （GSH） content increased 50.0-490.1 p.g g-~. GST activity of sensitive sugar beet decreased 122.6 U mg-~ min-1 compared with tolerant sugar beet and GSH content only increased to 7.4 p.g g-~ at chlorimuron-ethyl 0.5 g a.i. ha-~ in sensitive sugar beet. The higher GST activity and GSH content conjugated chlorimuron-ethyl absorbed in tolerant sugar beet and made it lost activity. Acetolactate synthases （ALS） activity of the tolerant sugar beet increased to 62.5 and 70.6%, respectively in seedling and leaf growth period, at the same time ALS activity of the sensitive variety was decreased to 36.8 and 64.8%, respectively. The rapidly increased GST activity, GSH content, and ALS, the target enzyme activity were the important pathways for enduring chlorimuron-ethyl in tolerant sugar beet.     

等位基因特异性扩增技术鉴定慈姑对磺酰脲类除草剂抗药性

以抗感磺酰脲类除草剂慈姑为材料,采用ASPCR技术鉴定ALS功能区第324位氨基酸突变位点.结果表明,抗性慈姑生态型ALS第324位点发生了丙氨酸替代苏氨酸（感性生态型）的突变.同时发现,抗性慈姑生态型ALS第197位点也发生了非脯氨酸的替代突变.说明简便的ASPCR技术对ALS基因单碱基抗性突变鉴定是可行的.     

Study on Mutations in ALS for Resistance to Tribenuron-Methyl in Galium aparine L.

In recent years,Galium aparine L.has not been controlled by tribenuron-methyl in major Chinese winter wheat fields.The objective of this study is to understand the molecular basis of the resistance mechanism to tribenuron-methyl in G.aparine and to find the specific mutation sites in amino acid sequence of acetolactate synthase(ALS)in the resistant biotype of G.aparine.Fragments that encode the ALS were amplified and cloned from susceptible(S)and resistant(R)biotypes of G.aparine to tribenuron-methyl and sequenced subsequently.The result showed that the nucleotide sequence of Rbiotype of G.aparine differed from that of the S biotype with three amino acid substitutions,of which,the amino acid substitution of Trp574(TGG)to Gly(GGG)is located in the highly conserved region Domain B.The substitution of Trp574might be responsible for the resistance to tribenuron-methyl in the R-biotype of G.aparine.     

Induction of nitric oxide-dependent apoptosis in motor neurons by zinc-deficient superoxide dismutase

Mutations in copper, zinc superoxide dismutase (SOD) have been implicated in the selective death of motor neurons in 2 percent of amyotrophic lateral sclerosis (ALS) patients. The loss of zinc from either wild-type or ALS-mutant SODs was sufficient to induce apoptosis in cultured motor neurons. Toxicity required that copper be bound to SOD and depended on endogenous production of nitric oxide. When replete with zinc, neither ALS-mutant nor wild-type copper, zinc SODs were toxic, and both protected motor neurons from trophic factor withdrawal. Thus, zinc-deficient SOD may participate in both sporadic and familial ALS by an oxidative mechanism involving nitric oxide.     

机载激光雷达林区数字高程模型的建立

选择地处草原和森林过渡地带的上库力农场作为研究区(E120°36'50.48″~120°52'56.53″,N50°21'11.08″~50°24'32″),由机载激光雷达Leica ALS60采集实验数据,对Terra Solid分类获取的地形点建立数字高程模型(DEM);利用IDL编译一次样条有限元内插法对点云数据进行分块处理,分析生产DEM的精度。结果表明:1.0、1.5、2.0 m三种不同分辨率的DEM精度,分别为0.034、0.078、0.096 m。     

Induced pluripotent stem cells generated from patients with ALS can be differentiated into motor neurons

The generation of pluripotent stem cells from an individual patient would enable the large-scale production of the cell types affected by that patient's disease. These cells could in turn be used for disease modeling, drug discovery, and eventually autologous cell replacement therapies. Although recent studies have demonstrated the reprogramming of human fibroblasts to a pluripotent state, it remains unclear whether these induced pluripotent stem (iPS) cells can be produced directly from elderly patients with chronic disease. We have generated iPS cells from an 82-year-old woman diagnosed with a familial form of amyotrophic lateral sclerosis (ALS). These patient-specific iPS cells possess properties of embryonic stem cells and were successfully directed to differentiate into motor neurons, the cell type destroyed in ALS.     

山东省小麦田播娘蒿对双氟磺草胺抗性水平及靶标抗性机理

【背景】播娘蒿（Descurainia sophia）是一种冬小麦田分布广泛且危害严重的阔叶杂草,双氟磺草胺则是目前冬小麦田防除阔叶杂草应用面积最大的一种ALS抑制剂类除草剂。双氟磺草胺经多年应用后,对部分区域冬小麦田播娘蒿防治效果下降,可能与当地播娘蒿对双氟磺草胺产生抗药性有关。【目的】明确山东省冬小麦田播娘蒿对双氟磺草胺的抗性水平和抗性机理,为制定小麦田播娘蒿等阔叶杂草精准区域防控提供理论依据。【方法】以播娘蒿为研究对象,在温室内采用整株生物测定法测定40个播娘蒿种群对双氟磺草胺和对比药剂苯磺隆、2甲4氯共3种除草剂的抗性水平,同时根据播娘蒿ALS基因序列,设计引物,提取对双氟磺草胺高抗的播娘蒿单株基因组DNA,测序获得序列与敏感型基因进行比对,查找突变位点,明确靶标抗性机理。【结果】抗性水平测定结果表明,40个播娘蒿种群中有32个对双氟磺草胺敏感,占80.00%,低抗、中抗和高抗种群分别有3、3和2个,JN-1、JNI-2、LY-2为低抗种群,LC-3、LY-4、YT-1为中抗种群,相对抗性指数（RI）分别为49.00、26.44、21.09,BZ-1和DZ-3属于高抗种群,RI分别为52.00和194.00。对BZ-1和DZ-3进行的ALS基因检测结果表明,BZ-1种群ALS基因第197位氨基酸发生CCT（Pro）到TCT（Ser）或CTT（Leu）突变,DZ-3种群ALS基因第574位氨基酸发生TGG（Trp)到TTG（Leu）突变。另外,40个种群中对对比药剂苯磺隆产生抗性的种群有19个,占总样点数的47.50%,低抗种群有11个,中抗种群有6个,RI分别为38.05、13.55、11.54、10.45、11.50、11.02,高抗种群为DZ-3和LY-4,RI分别为244.75和68.50;40个播娘蒿种群对另一种对比药剂2甲4氯没有产生抗药性。【结论】山东省小麦田采集的40个播娘蒿种群中已有20.00%的种群对双氟磺草胺产生抗性,且高抗双氟磺草胺的播娘蒿种群发生不同位置氨基酸取代。播娘蒿对苯磺隆的抗性仍很严重,但对激素类除草剂2甲4氯未产生抗药性。针对播娘蒿发生区域,不能单一使用双氟磺草胺,应推广多种作用机理的除草剂交替、混和使用,从而延缓和控制杂草产生抗药性,同时扩大杀草谱、降低除草剂使用量。     

山东省小麦田大穗看麦娘抗性水平、靶标抗性机理及田间防除效果测定

【背景】大穗看麦娘（Alopecurus myosuroides）是我国大陆近几年新发展蔓延的一种恶性禾本科杂草,目前已在山东、河南、河北、安徽等省有分布,且分布面积不断扩大。【目的】明确山东省冬小麦田大穗看麦娘对常用除草剂的抗性水平及部分种群产生抗性的机理,并评价不同除草剂对其田间防除效果,为制定小麦田大穗看麦娘防控技术规程提供理论依据。【方法】室内采用整株生物测定法测定9个大穗看麦娘种群对乙酰乳酸合成酶（ALS）抑制剂除草剂啶磺草胺、甲基二磺隆以及乙酰辅酶A羧化酶（ACC）抑制剂除草剂唑啉草酯、炔草酯、精噁唑禾草灵共5种除草剂的抗性水平,并对产生抗性的种群进行靶标基因检测,同时分别在冬前和冬后开展田间试验,评价不同除草剂对大穗看麦娘的田间防除效果。【结果】室内测定结果表明,种群8（JN2）对啶磺草胺和甲基二磺隆产生明显抗性,相对抗性指数分别达到47.32、15.97,靶标基因检测显示,该种群内植株ALS基因编码的第197位点氨基酸发生由脯氨酸（CCC）到苏氨酸（ACC）的突变;所有种群对唑啉草酯、炔草酯和精噁唑禾草灵均表现敏感。田间试验结果表明,冬前使用除草剂对大穗看麦娘的防除效果优于冬后使用除草剂的效果,冬前唑啉草酯对大穗看麦娘鲜重防效为98.6%,而冬后处理为89.1%;冬前使用啶磺草胺、甲基二磺隆对大穗看麦娘的株防效和鲜重防效在72.2%—89.3%,冬后使用则为68.6%—83.2%;唑啉草酯、炔草酯和精噁唑禾草灵对大穗看麦娘均表现出很好的防除效果,冬前使用的株防效和鲜重防效均在96.2%以上,冬后使用在82.6%—92.2%。【结论】供试的9个大穗看麦娘种群中,发现1个种群对甲基二磺隆、啶磺草胺产生较高抗性,但未发现对唑啉草酯等产生抗性的种群,室内试验和田间试验结果具有一致性。