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In determining the structure of large proteins by NMR, it would be desirable to obtain complete backbone, side-chain, and NOE assignments efficiently, with a minimum number of experiments and samples. Although new strategies have made backbone assignment highly efficient, side-chain assignment has remained more difficult. Faced with the task of assigning side-chains in a protein with poor relaxation properties, the Tetrahymena histone acetyltransferase tGCN5, we have developed an assignment strategy that would provide complete side-chain assignments in cases where fast 13C transverse relaxation causes HCCH-TOCSY experiments to fail. Using the strategy presented here, the majority of aliphatic side-chain proton and carbon resonances can be efficiently obtained using optimized H(CC-CO)NH-TOCSY and (H)C(C-CO)NH-TOCSY experiments on a partially deuterated protein sample. Assignments can be completed readily using additional information from a 13 C-dispersed NOESY-HSQC spectrum. Combination of these experiments with H(CC)NH-TOCSY and (H)C(C)NH-TOCSY may provide complete backbone and side-chain assignments for large proteins using only one or two samples.  相似文献   
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Shi L  Lu Y  Sun J  Zhang J  Sun C  Liu J  Shen J 《Biomacromolecules》2003,4(5):1161-1167
We describe here a new approach to construct a multilayer enzyme/polyelectrolyte film on a structured transparent indium-tin oxide (ITO) covered glass electrode surface as micropattern, on which two different types of enzyme distributed laterally on one common substrate without interference. The multilayer film was prepared by alternate electric field directed layer-by-layer assembly deposition and alternate deposition of different redox enzymes and polyelectrolyte poly(diallyldimethylammonium chloride) (PDDA) onto the site-selective ITO glass electrode surface. The cyclic voltammogram, obtained from the ITO glass electrode modified with the glucose oxidase (GO(X))/PDDA and catalase (CA(T))/PDDA multilayers, revealed that the bioelectrocatalytic response is directly correlated to the number of deposition bilayers. From the analysis of cyclic voltammetric characterization, the coverage of catalytically active enzymes per enzyme/PDDA bilayer during the multilayer formation was homogeneous, which demonstrates that the multilayer is constructed in a spatially ordered manner. Also, from the atomic force microscopy and Brewster angle microscopy measurements, more information of the multilayer constructed by different methods on the modified electrode surface is obtained and compared. This fabrication technique is simple and would be applicable to the construction of a thickness- and area-controlled biopattern composed of multi-enzymes as well as multiple biomaterials.  相似文献   
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We report the results of a genetic screen to identify molecules important for synapse formation and/or maintenance. siRNAs were used to decrease the expression of candidate genes in neurons, and synapse development was assessed. We surveyed 22 cadherin family members and demonstrated distinct roles for cadherin-11 and cadherin-13 in synapse development. Our screen also revealed roles for the class 4 Semaphorins Sema4B and Sema4D in the development of glutamatergic and/or GABAergic synapses. We found that Sema4D affects the formation of GABAergic, but not glutamatergic, synapses. Our screen also identified the activity-regulated small GTPase Rem2 as a regulator of synapse development. A known calcium channel modulator, Rem2 may function as part of a homeostatic mechanism that controls synapse number. These experiments establish the feasibility of RNAi screens to characterize the mechanisms that control mammalian neuronal development and to identify components of the genetic program that regulate synapse formation and/or maintenance.  相似文献   
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Li  Luhua  An  Chang  Wang  Zhongni  Xiong  Fumin  Wang  Yingxi  Ren  Mingjian  Xu  Ruhong 《Plant Growth Regulation》2022,98(1):127-139
Plant Growth Regulation - Anthocyanidin synthase (ANS) is involved in the synthesis of anthocyanins, which are important phytonutrients because of their beneficial effects on human health. Here, we...  相似文献   
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Objectives

The objective of this study was to identify the risk factors for ARI in the Australian community.

Methods

We used a national survey of 7578 randomly selected respondents in 2008–2009 to identify the risk factors of ARI. A case was defined as a person experiencing cold or flu with one or more symptoms of: fever, chills, sore throat, runny nose, or cough in the previous four weeks.

Results

There were 19.8% (1505/7578) of respondents who reported ARI in the four weeks prior to the survey. Age was an independent risk factor for ARI, with the risk of acquiring ARI decreasing as age increased. Respondents reporting asthma (OR 1.4, 95%CI: 1.2–1.7) or having someone in their house attending childcare (OR 1.6, 95%CI: 1.2–2.1) were more likely to report ARI.

Conclusions

It is important to identify ways of interrupting transmission of ARI amongst children. Improving identification of risk factors will enable targeted interventions for this exceedingly common syndrome.  相似文献   
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在整理保存于中国科学院植物研究所中国国家植物标本馆(PE)的国产合瓣花植物模式标本时,根据《国际藻类、菌类、植物命名法规》(墨尔本法规)规则9.5,发现一些植物名称的模式为合模式,其中包括杜鹃花科的奇异杜鹃、怒江杜鹃、绢毛杜鹃、橙黄杜鹃、光蕊杜鹃、棕背川滇杜鹃、拟镰果杜鹃、镰果杜鹃、粘毛杜鹃、粗毛杜鹃、腾冲杜鹃、剑川杜鹃和红粗毛杜鹃。遵照规则8.1、9.11和9.12,以及辅则9A.3的精神,对这13个名称做出后选模式指定。  相似文献   
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Pharmacological stimulation of adipose tissue remodeling and thermogenesis to increase energy expenditure is expected to be a viable therapeutic strategy for obesity. Berberine has been reported to have pharmacological activity in adipose tissue to anti-obesity, while the mechanism remains unclear. Here, we observed that berberine significantly reduced the body weight and insulin resistance of high-fat diet mice by promoting the distribution of brown adipose tissue and thermogenesis. We have further demonstrated that berberine activated energy metabolic sensing pathway AMPK/SIRT1 axis to increase the level of PPARγ deacetylation, which leads to promoting adipose tissue remodeling and increasing the expression of the thermogenic protein UCP-1. These findings suggest that berberine that enhances the AMPK/SIRT1 pathway can act as a selective PPARγ activator to promote adipose tissue remodeling and thermogenesis. This study proposes a new mechanism for the regulation of berberine in adipose tissue and offers a great prospect for berberine in obesity treatment  相似文献   
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