Regulation of the Polycomb protein RING1B ubiquitination by USP7

The E3 ubiquitin ligase RING1B plays an important role in Polycomb-mediated gene silencing by monoubiquitinating histone H2A. Both the activity and stability of RING1B are controlled by ubiquitination in two distinct manners. Self ubiquitination of RING1B generates K6, K27 and K48-based mixed polyubiquitin chain, and is required for its activity as a ligase. On the other hand, its proteasomal degradation is mediated by another ligase; E6-AP catalyzes the formation of K48-based chains. Since these two modes of ubiquitination target the same lysine residues and are therefore mutually exclusive, an important mode of regulation of RING1B should be at the level of deubiquitination. Here we identify USP7 as a deubiquitinating enzyme that regulates the ubiquitination state of RING1B. RING1B interacts with USP7, which is mediated in part by its RING domain. In addition, USP7 was found in a complex with other Polycomb proteins, suggesting a broad role in regulating these complexes. Although, USP7 directly and specifically deubiquitinates RING1B in vitro and in vivo, it does not discriminate between the activating and proteolysis-targeting modes of ubiquitination, and therefore has a stabilizing effect on RING1B.     

Muscodor fengyangensis sp. nov. from southeast China: morphology, physiology and production of volatile compounds

The fungal genus Muscodor was erected on the basis of Muscodor albus, an endophytic fungus originally isolated from Cinnamomum zeylanicum. It produces a mixture of volatile organic compounds (VOCs) with antimicrobial activity that can be used as mycofumigants. The genus currently comprises five species. Here we describe the isolation and characterization of a new species of Muscodor on the basis of five endophytic fungal strains from leaves of Actinidia chinensis, Pseudotaxus chienii and an unidentified broad leaf tree in the Fengyangshan Nature Reserve, Zhejiang Province, Southeast of China. They exhibit white colonies on potato dextrose agar (PDA) media, rope-like mycelial strands, but did not sporulate. The optimum growth temperature is 25°C. The results of a phylogenetic analysis based on four loci (ITS1-5.8S-ITS2, 28S rRNA, rpb2 and tub1) are consistent with the hypothesis that these five strains belong to a single taxon. All five strains also produce volatile chemical components with antimicrobial activity in vitro, which were different from those previously described for other Muscodor species.     

Synthesis and analytical investigation of C‐terminally modified peptide aldehydes and ketone: application to oxime ligation

C‐terminally modified peptides aldehyde (glycinal and alpha‐oxo aldehyde peptides) and ketone (pyruvic acid‐containing peptide) were synthesised to get new insights into the mechanism of acido‐catalysed oxime ligation. Their tetrahedral hydrated forms were investigated in solution and in the gas phase, using NMR and in‐source collision‐induced dissociation mass spectrometry, respectively, and the kinetics of the oximation reactions followed using analytical HPLC. The results obtained confirmed that the first step of the oximation reaction was the limiting step for the pyruvic acid‐containing peptides because of the steric effect and of the carbon angular strain of the ketone. The second step is the determining step for the aldehyde peptides because the basicity of the oxygen of the hydroxyl function of the tetrahedral form is greater for glycinal than for alpha‐oxo aldehyde. These data strongly suggest that the hydrated form of the aldehyde partner has to be considered when oxime reactions are performed in aqueous buffer. Copyright © 2011 European Peptide Society and John Wiley & Sons, Ltd.     

Comprehensive proteome analysis in Cenococcum geophilum Fr. as a tool to discover drought-related proteins

Cenococcum geophilum is a widely distributed ectomycorrhizal fungus potentially playing a significant role in resistance and resilience mechanisms of its tree hosts exposed to drought stress. In this study, we performed a large scale protein analysis in pure cultures of C. geophilum in order to gain first global insights into the proteome assembly of this fungus. Using 1-D gel electrophoresis coupled with ESI-MS/MS, we indentified 638 unique proteins. Most of these proteins were related to the metabolic and cellular processes, and the transport machinery of cells. In a second step, we examined the influence of water deprivation on the proteome of C. geophilum pure cultures at three time points of gradually imposed drought. The results indicated that 12 proteins were differentially abundant in mycelia subjected to drought compared to controls. The induced responses in C. geophilum point towards regulation of osmotic stress, maintainance of cell integrity, and counteracting increased levels of reactive oxygen species formed during water deprivation.     

A comparative study of enol aldehyde formation from betamethasone, dexamethasone, beclomethasone and related compounds under acidic and alkaline conditions

Enol aldehydes are one type of key degradation and metabolic intermediates from a group of corticosteroids containing the 1,3-dihydroxyacetone side chain on their D-rings, such as betamethasone, dexamethasone, beclomethasone, and related compounds. The formation of enol aldehydes from these corticosteroids is via acid-catalyzed β-elimination of water from the side chain, a process known as Mattox rearrangement. It was recently reported by our group that enol aldehydes could also be formed directly from the corresponding 17,21-diesters of these corticosteroids but only under alkaline condition, which was proposed to follow a variation pathway of the original Mattox rearrangement. In this paper, we report the results of a comparative study of enol aldehyde formation from these structurally similar corticosteroids (under the original acidic Mattox condition) and their 17,21-diesters (under the alkaline Mattox variation condition), respectively. In general, enol aldehydes were found to be formed under both conditions; however, the ratios of the E- and Z-isomers of the enol aldehyde were different in each case. The only exception was beclomethasone 17,21-diester under the alkaline condition, where a competing elimination of HCl from the 9,11-positions became predominant. These results can be explained by their structural differences with regard to the Mattox mechanism and its variation pathway. Lastly, solvent effect under acidic condition was studied between an aprotic and a protic solvent and the result suggests that enol aldehyde formation is greatly favored in an aprotic environment.     

Improved lux reporters for use in Staphylococcus aureus

The use of luxABCDE (lux) offers certain advantages over other reporters, such as: lacZ and xylE. It is real time and its signal generation is produced without the requirement for any additional substrates. In some bacteria such as Staphylococcus spp, light production by luciferase is restricted because of a limited availability of endogenous substrates such as fatty acid aldehyde. We describe the construction of promoterless-lux cloning vectors, pGYlux and pAmilux. S. aureus carrying B. subtilis xyl/tetO promoter fused to the lux genes of pGYlux gave up to a 2.5-fold enhancement of luminescence over S. aureus carrying the xyl/tetO promoter fused to lux genes of the previously published parent vector pAL2. Furthermore, pAmilux showed a 6-fold enhancement of lux expression when compared to pGYlux in S. aureus. This was achieved by cloning the constitutive ami promoter upstream of the luxCDE genes to increase endogenous fatty acid aldehyde production while maintaining its reporter functionality by fusing promoters to the luxAB genes.     

Acrolein scavenging: a potential novel mechanism of attenuating oxidative stress following spinal cord injury

It has long been established that oxidative stress plays a critical role in the pathophysiology of spinal cord injury, and represents an important target of therapeutic intervention following the initial trauma. However, free radical scavengers have been largely ineffective in clinical trials, and as such a novel target to attenuate oxidative stress is highly warranted. In addition to free radicals, peroxidation of lipid membranes following spinal cord injury (SCI) produces reactive aldehydes such as acrolein. Acrolein is capable of depleting endogenous antioxidants such as glutathione, generating free radicals, promoting oxidative stress, and damaging proteins and DNA. Acrolein has a significantly longer half‐life than the transient free radicals, and thus may represent a potentially better target of therapeutic intervention to attenuate oxidative stress. There is growing evidence, from our lab and others, to suggest that reactive aldehydes such as acrolein play a critical role in oxidative stress and SCI. The focus of this review is to summarize the cellular and biochemical mechanisms of acrolein‐induced membrane damage, mitochondrial injury, oxidative stress, cell death, and functional loss. Evidence will also be presented to suggest that acrolein scavenging may be a novel means of therapeutic intervention to attenuate oxidative stress and improve recovery following traumatic SCI.     

实验性大鼠溃疡性结肠炎菌群失调的诊断及复方树舌的治疗观察

目的探讨对溃疡性结肠炎厌氧菌菌群失调简便检查方法,寻找有效治疗溃疡性结肠炎的中药合剂。方法用冰乙酸诱发大鼠结肠炎模型,用复方树舌中药治疗,在治疗期间分别检测每组大鼠厌氧菌代谢产物、挥发性脂肪酸的含量、了解肠厌氧菌群改变,一般切片行HE染色,观察病理变化。结果（1）模型组大鼠挥发性脂肪酸明显低于正常组（P〈0．05）;pH从第2天起升高明显高于正常对照组（P〈0．05）,各治疗组与自然恢复组比较差异有显著性（P〈0．05）。（2）自然恢复组与各用药组间的炎症评分比较P〈0．01,各治疗组间比较,树舌加丽珠肠乐组组织损伤评分小（P〈0．05）。结论（1）测定厌氧菌的代谢产物挥发性脂肪酸含量,即可对肠内厌氧菌菌群失调做出判断。（2）实验证明,复方树舌中药可作为益生元调节肠道菌群失调及黏膜的修复,对溃疡性结肠炎疗效显著。     

Age-Dependent Variations of Volatile Emissions and Inhibitory Activity Toward <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in Tomato Leaves Treated with Chitosan Oligosaccharide

We investigated variations in the level and composition of volatiles emitted by tomato leaves at different ages. Our focus also included their antifungal properties and responses to chitosan oligosaccharide. Based on leaf position, the release of volatiles decreased over time. Young leaves produced high levels of C6-aldehyde, which is mainly composed of hexenal, while the volatiles emitted by more mature leaves largely comprised terpenes, particularly β-phellandrene and caryophyllane. In young upper leaves, the main components (up to 86% of the total) were hexenal, β-phellandrene, and caryophyllane. Their levels decreased steadily over time, from 386.3 μg g⁻¹ fresh weight (FW) in young leaves to 113.2 μg g⁻¹ FW in old tissues. Volatiles emitted from young leaves exhibited the best antifungal activity against spore germination and hyphal growth by Botrytis cinerea and Fusarium oxysporum. Leaves became more susceptible to oligosaccharide treatment with increasing age. When young tissues were exposed to chitosan, we found declines in both the quantity of volatiles and their ability to inhibit fungal growth. Compared with the control, the amount of volatiles from young tissues was 88.4% lower after such treatment. In contrast, contents of volatiles from old and adult leaves were dramatically increased by chitosan oligosaccharide. Likewise, their inhibitory effect was significantly enhanced. Therefore, our results suggest that these volatiles are responsible for antifungal activity and may play a role in age-related resistance by tomato.     

Biofiltration of composting gases using different municipal solid waste-pruning residue composts: monitoring by using an electronic nose

The concentration of volatile organic compounds (VOCs) during the composting of kitchen waste and pruning residues, and the abatement of VOCs by different compost biofilters was studied. VOCs removal efficiencies greater than 90% were obtained using composts of municipal solid waste (MSW) or MSW-pruning residue as biofilter material. An electronic nose identified qualitative differences among the biofilter output gases at very low concentrations of VOCs. These differences were related to compost constituents, compost particle size (2-7 or 7-20 mm), and a combination of both factors. The total concentration of VOCs determined by a photoionization analyser and inferred from electronic nose data sets were correlated over an ample range of concentrations of VOCs, showing that these techniques could be specially adapted for the monitoring of these processes.