Identification of small molecules that modify the protein folding activity of heat shock protein 70

Molecular chaperones, such as heat shock protein 70 (Hsp70) and its bacterial ortholog DnaK, play numerous important roles in protein folding. In vitro, this activity can be observed by incubating purified chaperones with denatured substrates and measuring the recovery of properly folded protein. In an effort to rapidly identify small molecules that modify this folding activity, we modified an existing method for use in 96-well plates. In this assay, denatured firefly luciferase was treated with a mixture of DnaK and prospective chemical modulators. The luminescence of refolded luciferase was used to follow the reaction progress, and counterscreens excluded compounds that target luciferase; thus, hits from these screens modify protein folding via their effects on the function of the chaperone machine. Using this platform, we screened a pilot chemical library and found five new inhibitors of DnaK and one compound that promoted folding. These chemical probes may be useful in studies aimed at understanding the many varied roles of chaperones in cellular protein folding. Moreover, this assay provides the opportunity to rapidly screen for additional compounds that might regulate the folding activity of Hsp70.     

Effects of media composition on submerged culture spores of the entomopathogenic fungus,Metarhizium anisopliae var. acridum Part 2: Effects of media osmolality on cell wall characteristics,carbohydrate concentrations,drying stability,and pathogenicity

This study evaluates osmolality of a submerged conidia-producing medium in relation to the following spore characteristics: yield, morphology (dimensions and cell wall structure), chemical properties of cell wall surfaces (charge, hydrophobicity, and lectin binding), cytoplasmic polyols and trehalose, and performance (drying stability and pathogenicity). Spore production was increased by the addition of up to 150 g l^?1 polyethylene glycol 200 (PEG). Spores from high osmolality medium (HOM spores) containing 100 g l^?1 PEG had thin cell walls and dimensions more similar to blastospores than submerged conidia or aerial conidia. However, a faint electron-dense layer separating primary and secondary HOM spores’ cell walls was discernable by transmission electron microscopy as found in aerial and submerged conidia but not found in blastospores. HOM spores also appeared to have an outer rodlet layer, unlike blastospores, although it was thinner than those observed in submerged conidia. HOM spores’ surfaces possessed hydrophobic microsites, which was further evidence of the presence of a rodlet layer. In addition, HOM spores had concentrations of exposed N-acetyl-β-d-glucosaminyl residues intermediate between blastospores and submerged conidia potentially indicating a masking of underlying cell wall by a rodlet layer. All spore types had exposed α-d-mannosyl and/or α-d-glucosyl residues, but lacked oligosaccharides. Similar to blastospores, HOM spores were less anionic than submerged conida. Although HOM spores had thin cell walls, they were more stable to drying than blastospores and submerged conidia. Relative drying stability did not appear to be the result of differences in polyol or trehalose concentrations, since trehalose concentrations were lower in HOM spores than submerged conidia and polyol concentrations were similar between the two spore types. HOM spores had faster germination rates than submerged conidia, similar to blastospores, and they were more pathogenic to Schistocerca americana than submerged conidia and aerial conidia.     

西洋参愈伤组织悬浮培养基质中蔗糖和无机元素的消耗状况

植物离体培养所采用的基本基质大多数是广谱性的,各种营养成分是否被充分利用因植物种类不同而异,这方面的研究报道较少[1～3],尤其在西洋参(PanaxquinquefoliumLinn.)芽胞愈伤组织悬浮培养中尚未见报道。本实验通过对西洋参愈伤组织培养基中蔗糖和无机元素消耗的研究,确定培养周期中碳源和无机元素的用量,为西洋参大规模细胞培养中碳源和无机元素及时补充和起始供给浓度的确定提供参数。1　材料与方法1.1　供试材料与培养条件采用本实验室在1999年筛选的西洋参芽胞愈伤组织无性系作为实验材料。先将固体培养基上的愈伤组织转入含有20m…     

蛋白激酶B的PH结构域可溶性表达与纯化及其二级结构分析

蛋白激酶B(亦称为Akt)是一种蛋白质丝氨酸 苏氨酸激酶 ,因其与蛋白激酶A(PKA)和蛋白激酶C(PKC)具有相对高的同源性 ,而被命名为PKB .作为磷脂酰肌醇 3激酶 (PI3 kinase)的下游分子 ,它广泛参与细胞各种功能的调节 .PKB对代谢的影响主要表现为促进蛋白质的合成 ,促进糖原的转运[1] .同时 ,PKB还在细胞增殖与调控中发挥重要作用[2 ] .PH结构域 (pleckstrinhomologydomain)是一种存在于多种信号蛋白质和细胞骨架相关蛋白质中的功能性区域 .它通常由 7个反向平行的 β片层和C末端的一个α螺旋构成 .PH结构域的配体具有一定的多样性…     

Jasmonate effects on in vitro tuberization and tuber bulking in two potato cultivars (Solanum tuberosum L.) under different media and photoperiod conditions

Summary Jasmonic acid (JA) effects on in vitro tuberization of potato nodal explants cvs. Sangre and Russet Burbank were tested under liquid and solid media conditions and 0,8, and 16h photoperiod. Explants taken from stock plants grown on 2.5μM JA-supplemented medium tuberized first, particularly in darkness. The most pronounced benefits of the JA pretreatment were recorded under 16h photoperiod, which is known to inhibit tuberization. Cultivar Sangre benefited from the JA preconditioning of stock plants more than Russet Burbank. Russet Burbank required the JA supplement in tuberization media to reach the same degree of stimulation. Overall, microtubers produced either from JA preconditioned stock plants or on the JA-containing tuberization media were more uniform and larger than from other treatments. Eight hours photoperiod was by far the best treatment for the production of high-quality uniform microtubers. JA conditioning of stock plants prior to taking explants for tuberization is being proposed as a treatment enhancing the quality of microtubers.     

Effect of Manipulations on the Host Selection Behavior of Sitobion avenae (Homoptera: Aphididae)

The study of aphid host selection and feeding behavior is difficult because aphids have to penetrate the plant to reach their feeding site, phloem tissue. The activity of the stylets, salivation or food intake, can not be observed externally and requires an indirect visualization technique such as the Electric Penetration Graph (EPG). The plant selection behavior of Sitobion avenae on potato varied depending on whether an ethological or EPG method was used to study it. A similar variation did not occur with Myzus persicae or Rhopalosiphum padi. The application of water-based silver conductive paint onto the thorax, as normally used for EPG, or onto the abdomen of Sitobion avenae alates resulted in increased duration and frequency of probing compared to results from ethological observations. Our results indicated that EPG manipulations might have different effects on different species of aphids and that a comparison of EPG and ethological data is required to confirm that the EPG method does not bias aphid feeding behavior.     

Enantiomers of a nonylphenol isomer: absolute configurations and estrogenic potencies

Enantiomers of 4-(1,1,2-trimethylhexyl)phenol, a chiral isomer of the endocrine disrupting chemical nonylphenol, have been resolved and isolated by preparative chiral HPLC. The absolute configurations of the enantiomers were then determined by an X-ray crystallographic study of the (-)-camphanoyl derivative of the first eluted enantiomer NP(35)E1. The first enantiomer (NP(35)E1) and the second enantiomer (NP(35)E2) eluted were found to have the S and R absolute configurations, respectively. The estrogenic potencies of the S and R enantiomers were tested by the E-screen assay. A slight difference was observed in the relative proliferative effect between the S enantiomer and R enantiomer in the E-screen assay.     

电子健康档案管理探讨

电子健康档案管理以健康需求为导向,以电子健康档案为基础,以电子网络为依托提供个性化的健康服务。在新医改的大背景下,电子健康档案管理通过“早预防、早诊断、早治疗”,降低医疗费用,提高人们的生活质量,营造和谐的社会氛围,贯彻落实科学发展观,以适应全面建设小康社会的需求。     

Spent mushroom substrates as component of growing media for germination and growth of horticultural plants

This research work was conducted in order to investigate the possibility of using spent mushroom substrate (SMS) in the production of horticultural seedlings replacing part of the peat in the growing media. Three vegetable species with different salt sensitivities, the less sensitive being tomato (Lycopersicon esculentum var. Muchamiel), the moderately salt-sensitive being courgette (Cucurbita pepo L. var. Afrodite F1) and the most salt-sensitive being pepper (Capsicum annum L. var. Lamuyo F1) were grown in 12 media containing SMS of two types of mushroom (Agaricus bisporus (SMS-AB) and Pleurotus ostreatus (SMS-PO)) or a mixture of both 50% (v/v) (SMS-50), as well as peat in various ratios. The proportions of each residue in the mixtures elaborated with peat were 25%, 50%, 75% and 100% v/v residue. A substrate of 100% peat was used as control. The experiment was arranged in a completely-randomised design with two replicates per treatment under greenhouse conditions. Prior to sowing, some physical, physico-chemical and chemical properties of the growing media were determined and seed germination and fresh weight of seedling were also measured. In most of the cases, the addition of SMS to the growing media produced an increase in the pH values, salt contents, macro and micronutrient concentrations and a decrease in the water holding capacity contents in comparison to peat, whereas great differences were found in the air capacity values between SMS-based substrates and peat. Up to 75% SMS can be used in mixtures with peat for seed germination of the plant species studied. Regarding the most suitable SMS-based substrates for plant growth, any substrate could be used for tomato seedling production. However, all SMS-AB-based substrates and the media containing low dose of SMS-PO and SMS-50 were adequate for growth of courgette and pepper.     

Follow the money: international donors,external homelands and their effect on Romani media and advocacy

ABSTRACT

While Roma are both Europe’s largest minority, there is no “homeland” state that claims to guard their interest. The lack of “an external national homeland” [Brubaker, R. 1996. Nationalism Reframed: Nationhood and the National Question in the New Europe. Cambridge: Cambridge University Press] to watchdog and safeguard their rights has a particular effect on how Roma engage as political actors and subjects. International donors/foundations have assumed the role of “external homeland”. This article explores the effect that donors’ funding priorities have on Romani advocacy – specifically Romani journalism. Drawing upon multi-sited fieldwork in five countries, extensive document analysis, and interviews, Idemonstrate that the change in Romani media content over the past two decades reflects the shift in funders’ priorities, particularly in relation to the European project. “The Roma” become a tool for donors and European institutions to build a “European” identity while Romani-led advocacy becomes increasingly marganilized.