Detection of covalent enzyme-substrate complexes of nitrilase by ion-spray mass spectroscopy

Nitrilase from Rhodococcus ATCC 39484 was found to consist of two species of Mr 40,258 +/- 2 and 40,388 +/- 2 Da. When the enzyme was incubated with nitrile substrates and the reaction quenched with acid, higher Mr species were observed. The mass differences were consistent with addition of a substrate molecule to each species. These results represent the first reported demonstration that this, or any other nitrilase forms a covalent intermediate with its substrates. The observation that the intermediate, suggested to be either a thioimidate or an acylenzyme, can be trapped by acidification indicates that the rate of breakdown of the intermediate is rate-limiting.     

Pyrophosphate contamination in crystalline phosphocreatine: a source of error in kinetic studies using coupled enzymes.

Commercial crystalline phosphocreatine generally contains a contaminant, probably pyrophosphate, that can inhibit nucleoside triphosphatases and may therefore cause errors in steady-state kinetic studies of creatine kinase-coupled r reactions. Ion-exchange chromatography reveals the presence of pyrophosphate in some batches of phosphocreatine sufficient to account for the observed inhibition. Chromatographically purified phosphocreatine shows no inhibitory effect.     

高原湖泊流域国土空间生态修复优先区诊断及修复研究

高原湖泊流域是高原地区人类活动的重要载体,兼具高生态价值和高脆弱性的特点。随着高原湖泊流域城市化和工业化发展加速,湖泊面积萎缩,污染加剧,流域生态环境受损严重,引发了一系列生态环境问题,如水土流失、水污染、湿地退化、生境质量下降等。亟需开展生态修复以平衡经济发展与生态环境保护之间关系,而基于整体保护与系统治理思维诊断并修复生态修复优先区,是科学有序推进国土空间生态保护与修复的重要抓手。基于此,研究以高原湖泊流域典型代表滇池流域为例,利用人类足迹和景观生态风险模型定量评估生态系统所受负向干扰,以最小累积阻力模型和电路理论构建流域生态网络;提取生态网络受负向干扰较高的关键区域为生态修复优先区并提出针对性修复措施。研究表明：(1)滇池流域人类干扰和生态风险整体较高,人类干扰整体呈核心—边缘递减的圈层式分布,中高生态风险占据了绝大部分区域。人类交通网络大幅扩展了人类干扰和生态风险的强度和深度;(2)区域生态网络呈典型湖泊生态网络特点,38条生态廊道呈放射状或环状分布,连通湖区、山区两大生态空间内共23块生态源地,保障区域生态安全;(3)研究共提取生态源地修复优先区73.83km²     

中国森林生态系统林冠层降雨截留特征

森林生态系统作为陆地生态系统的主体,其发达的林冠层通过调节降水量、改变降水强度等深刻影响着流域全过程水文通量及水分输出。以中国广泛开展的典型森林降雨再分配过程的年尺度监测数据为基础,揭示中国不同类型森林生态系统的降雨再分配及林冠层降雨截留特征,阐明森林生态系统林冠层截留特征与降雨、植被要素的关系。结果表明:我国不同森林生态系统年穿透雨量处于141.4-2450.0 mm之间,年穿透雨率为36.3%-92.3%。5种典型森林生态系统多年平均穿透雨量((445.3±252.9)-(1230.6±479.6) mm)占同期多年平均降雨量的(72.6±9.2)%-(77.4±8.9)%。不同森林生态系统年树干茎流量介于0-508.2 mm之间,占同期年降雨量的0-25.8%。5种典型森林生态系统树干茎流量多年平均值((9.8±17.3)-(87.8±81.6) mm)占同期多年平均降雨量的(1.4±1.9)%-(5.4±4.6)%。不同森林生态系统林冠层年降雨截留范围在25.7-812.9 mm之间,占年降雨量的4.2%-55.6%。5种典型森林生态系统多年平均林冠截留量((154.2±81.6)-(392.2±203.5) mm)占同期年平均降雨量的(18.7±7.4)%-(25.9±8.3)%。进一步分析表明,我国森林生态系统穿透雨量、树干茎流量和林冠层截留量随观测区年降雨量的增加而呈显著增大(P<0.05),年穿透雨率、年树干茎流率随年降雨量的增加呈显著线性上升趋势(P<0.05),而年林冠截留率与年降雨量呈显著的负相关关系(P<0.01),降雨量、叶面积指数是深刻影响森林生态系统林冠层降雨截留率等特征的重要因素。整体上,不同类型森林生态系统林冠截留降雨能力存在明显差异,林冠层截留率突出表现为:落叶林大于常绿林、针叶林大于阔叶林。     

3年高氮添加对黄山松土壤真菌网络稳定性的影响

土壤真菌群落对维持生态系统功能至关重要,然而氮沉降等环境变化如何影响其稳定性尚不清楚。利用高通量测序和网络分析,探究了连续三年不同氮添加水平对土壤真菌群落特征的影响,并通过计算"内聚力"量化连通性以预测真菌网络的稳定性。结果表明,真菌多样性及其群落稳定性对氮添加的响应存在明显的时间效应。相比于前2年,第3年氮添加改变了真菌β多样性,且低氮添加显著提高了0-10 cm土层真菌Chao1和Shannon指数。网络分析发现,随着氮添加量的增加真菌网络正连接比例增加,平均聚类系数和模块性降低。内聚力分析表明,第3年高氮添加显著降低了两个土层的真菌负:正内聚力的绝对值,而低氮添加提高或不改变两者的比值。这说明高剂量氮添加将通过破坏真菌群落的稳定性影响其生态系统服务功能。研究为评价不同水平氮添加下森林土壤真菌群落稳定性提供了新的见解,对评价氮沉降的生态效应具有科学意义。     

DAZL regulates proliferation of human primordial germ cells by direct binding to precursor miRNAs and enhances DICER processing activity

Understanding the molecular and cellular mechanisms of human primordial germ cells (hPGCs) is essential in studying infertility and germ cell tumorigenesis. Many RNA-binding proteins (RBPs) and non-coding RNAs are specifically expressed and functional during hPGC developments. However, the roles and regulatory mechanisms of these RBPs and non-coding RNAs, such as microRNAs (miRNAs), in hPGCs remain elusive. In this study, we reported a new regulatory function of DAZL, a germ cell-specific RBP, in miRNA biogenesis and cell proliferation. First, DAZL co-localized with miRNA let-7a in human PGCs and up-regulated the levels of >100 mature miRNAs, including eight out of nine let-7 family, miR21, miR22, miR125, miR10 and miR199. Purified DAZL directly bound to the loops of precursor miRNAs with sequence specificity of GUU. The binding of DAZL to the precursor miRNA increased the maturation of miRNA by enhancing the cleavage activity of DICER. Furthermore, cell proliferation assay and cell cycle analysis confirmed that DAZL inhibited the proliferation of in vitro PGCs by promoting the maturation of these miRNAs. Evidently, the mature miRNAs up-regulated by DAZL silenced cell proliferation regulators including TRIM71. Moreover, DAZL inhibited germline tumor cell proliferation and teratoma formation. These results demonstrate that DAZL regulates hPGC proliferation by enhancing miRNA processing.     

Phosphorus Regulates the Level of Signaling Molecules in Rice to Reduce Cadmium Toxicity

Phosphorus treatment can reduce Cd accumulation and Cd toxicity in rice, but alterations in the internal regulatory network of rice during this process have rarely been reported. We have removed the effect of cadmium phosphate precipitation from the hydroponic system, treated a pair of different Cd-response rice varieties with different levels of phosphorus and cadmium and examined the changes in physiological indicators and regulatory networks. The results demonstrated that phosphorus treatment significantly reduced Cd accumulation in both types of rice, although the antioxidant systems within the two types of rice produced opposite responses. Overall, 3 mM phosphorus treatment to Cd-N decreased the expression of OsIAA17 and OsACO1 by 32% and 37%, respectively, while increasing the expression of OsNR2 by 83%; these three genes regulate the synthesis of auxin, ethylene, and nitric oxide in rice. IAA and NO levels in rice shoots increased by 24% and 96%, respectively, and these changes contribute to Cd detoxification. The cadmium transporter genes OsHMA2, OsIRT1, and OsABCC1 were significantly down-regulated in Cd-N roots after triple phosphorus treatment. These data suggest that phosphorus treatment can reduce Cd accumulation and enhance Cd resistance in rice by affecting the expression of signaling molecules.     

Acoustic signal dominance in the multimodal communication of a nocturnal mammal

Multimodal communication in animals is common, and is particularly well studied in signals that include both visual and auditory components. Multimodal signals that combine acoustic and olfactory components are less well known. Multimodal communication plays a crucial role in agonistic interactions in many mammals, but relatively little is known about this type of communication in nocturnal mammals. Here, we used male Great Himalayan leaf-nosed bats Hipposideros armiger to investigate multimodal signal function in acoustic and olfactory aggressive displays. We monitored the physiological responses (heart rate [HR]) when H. armiger was presented with 1 of 3 stimuli: territorial calls, forehead gland odors, and bimodal signals (calls + odors). Results showed that H. armiger rapidly increased their HR when exposed to any of the 3 stimuli. However, the duration of elevated HR and magnitude of change in HR increased significantly more when acoustic stimuli were presented alone compared with the presentation of olfactory stimuli alone. In contrast, the duration of elevated HR and magnitude of change in HR were significantly higher with bimodal stimuli than with olfactory stimuli alone, but no significant differences were found between the HR response to acoustic and bimodal stimuli. Our previous work showed that acoustic and chemical signals provided different types of information; here we describe experiments investigating the responses to those signals. These results suggest that olfactory and acoustic signals are non-redundant signal components, and that the acoustic component is the dominant modality in male H. armiger, at least as it related to HR. This study provides the first evidence that acoustic signals dominate over olfactory signals during agonistic interactions in a nocturnal mammal.