Pathological effects of the mushroom toxin alpha-amanitin on BALB/c mice

The pathological effects of alpha-amanitin on BALB/c mice after receiving intravenous injection were evaluated by RP-HPLC and mouse genome oligonucleotide microarray. The content of alpha-amanitin in Amanita virosa was about 2833.8 microg/g dry fruiting body. The liver and kidneys showed critical pathological changes after alpha-amanitin poisoning, and sera BUN, Crea, ALT, AST, TBIL and DBIL were the sensitive markers. The compound alpha-amanitin was detected in liver and kidney tissue homogenates by RP-HPLC after 48 h. The results of mouse genome oligonucleotide microarray showed 146 genes' expression changed, which formed the alternant network. The expression of 66 genes decreased, while 80 ones increased with more than two-fold differential expression after 48 h. The compound alpha-amanitin influenced not only RNA polymerase II, but also the expression of its associated genes. The application of mouse oligo chip provided valuable data for further understanding the biological properties and molecular pathogenesis of alpha-amanitin, also might be helpful for screening the curative drug for alpha-amanitin intoxication.     

Composition of the sheath produced by the green alga Chlorella sorokiniana

AIMS: To investigate the chemical characterization of the mucilage sheath produced by Chlorella sorokiniana. METHODS AND RESULTS: Algal mucilage sheath was hydrolysed with NaOH, containing EDTA. The purity of the hydrolysed sheath was determined by an ATP assay. The composition of polysaccharide in the sheath was investigated by high-performance anion-exchange chromatography with pulsed amperometric detection. Sucrose, galacturonic acid, xylitol, inositol, ribose, mannose, arabinose, galactose, rhamnose and fructose were detected in the sheath as sugar components. Magnesium was detected in the sheath as a divalent cation using inductively coupled argon plasma. The sheath matrix also contained protein. CONCLUSIONS: It appears that the sheath is composed of sugars and metals. Mucilage sheath contains many kinds of saccharides that are produced as photosynthetic metabolites and divalent cations that are contained in the culture medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on chemical characterization of the sheath matrix produced by C. sorokiniana.     

Future affective technology for autism and emotion communication

People on the autism spectrum often experience states of emotional or cognitive overload that pose challenges to their interests in learning and communicating. Measurements taken from home and school environments show that extreme overload experienced internally, measured as autonomic nervous system (ANS) activation, may not be visible externally: a person can have a resting heart rate twice the level of non-autistic peers, while outwardly appearing calm and relaxed. The chasm between what is happening on the inside and what is seen on the outside, coupled with challenges in speaking and being pushed to perform, is a recipe for a meltdown that may seem to come ‘out of the blue’, but in fact may have been steadily building. Because ANS activation both influences and is influenced by efforts to process sensory information, interact socially, initiate motor activity, produce meaningful speech and more, deciphering the dynamics of ANS states is important for understanding and helping people on the autism spectrum. This paper highlights advances in technology that can comfortably sense and communicate ANS arousal in daily life, allowing new kinds of investigations to inform the science of autism while also providing personalized feedback to help individuals who participate in the research.     

Identification and characterization of multiple curcumin synthases from the herb Curcuma longa

Curcuminoids are pharmaceutically important compounds isolated from the herb Curcuma longa. Two additional type III polyketide synthases, named CURS2 and CURS3, that are capable of curcuminoid synthesis were identified and characterized. In vitro analysis revealed that CURS2 preferred feruloyl-CoA as a starter substrate and CURS3 preferred both feruloyl-CoA and p-coumaroyl-CoA. These results suggested that CURS2 synthesizes curcumin or demethoxycurcumin and CURS3 synthesizes curcumin, bisdemethoxycurcumin and demethoxycurcumin. The availability of the substrates and the expression levels of the three different enzymes capable of curcuminoid synthesis with different substrate specificities might influence the composition of curcuminoids in the turmeric and in different cultivars.     

Purification, characterization, and directed evolution study of a vitamin D3 hydroxylase from Pseudonocardia autotrophica

Vitamin D₃ (VD₃) is a fat-soluble prohormone that plays a crucial role in bone metabolism, immunity, and control of cell proliferation and cell differentiation in mammals. The actinomycete Pseudonocardia autotrophica is capable of bioconversion of VD₃ into its physiologically active forms, namely, 25(OH)VD₃ or 1α,25(OH)₂VD₃. In this study, we isolated and characterized Vdh (vitamin D₃ hydroxylase), which hydroxylates VD₃ from P. autotrophica NBRC 12743. The vdh gene encodes a protein containing 403 amino acids with a molecular weight of 44,368 Da. This hydroxylase was found to be homologous with the P450 belonging to CYP107 family. Vdh had the same ratio of the V_max values for VD₃ 25-hydroxylation and 25(OH)VD₃ 1α-hydroxylation, while other enzymes showed preferential regio-specific hydroxylation on VD₃. We characterized a collection of Vdh mutants obtained by random mutagenesis and obtained a Vdh-K1 mutant by the combination of four amino acid substitutions. Vdh-K1 showed one-order higher VD₃ 25-hydroxylase activity than the wild-type enzyme. Biotransformation of VD₃ into 25(OH)VD₃ was successfully accomplished with a Vdh-expressed recombinant strain of actinobacterium Rhodococcus erythropolis. Vdh may be a useful enzyme for the production of physiologically active forms of VD₃ by a single cytochrome P450.     

不同压缩程序对海量生物信息数据压缩效率的比较分析

海量生物信息数据的不断涌现迫切需要在数据压缩技术方面进行更多研究,以减轻服务器存储压力和提高网络传输及数据分析的效率。目前虽然已开发出大量数据压缩软件,但对于海量生物信息数据而言,应该选用何种软件和方法进行数据压缩,尚缺乏详细的综合比较分析。本文选择生物信息学领域中GenBank数据库中的典型核酸和蛋白质序列数据库以及典型生物信息软件Blast和EMBOSS为例,采用不同数据压缩软件进行综合比较分析,结果发现经典压缩软件compress的总体压缩效率很高,除压缩比率可接受之外,其压缩时间相对其他软件而言显著减少,甚至比并行化的hzip2（pbzip2）和gzip（pigz）软件的时间还少很多,故可优先考虑使用。7-Zip软件虽然具有最高的压缩比率,但压缩过程十分耗时,可用于数据的长期储存;而采用bzip2、rar以及gzip等软件压缩的文件,虽然压缩比率较7-Zip的偏低,但压缩过程相对而言还比较快速。具体应用中推荐使用经典压缩软件compress以及并行化运行的pbzip2和pigz软件,三者可作为同时兼顾压缩比率和压缩时间的优选。     

In vitro molecular pattern classification via DNA-based weighted-sum operation

Recent progress in molecular computation suggests the possibility of pattern classification in vitro. Weighted sum is a primitive operation required by many pattern classification problems. Here we present a DNA-based molecular computation method for implementing the weighted-sum operation and its use for molecular pattern classification in a test tube. The weights of the classifier are encoded as the mixing ratios of the differentially labeled probe DNA molecules, which are competitively hybridized with the input-encoding target molecules to compute the decision boundary of classification. The computation result is detected by fluorescence signals. We experimentally verify the underlying weight encoding scheme and demonstrate successful discrimination of two-group labels of synthetic DNA mixture patterns. The method can be used for direct computation on biomolecular data in a liquid state.     

An arginyl residue in rice UDP-arabinopyranose mutase is required for catalytic activity and autoglycosylation

Plants use UDP-arabinofuranose (UDP-Araf) to donate Araf residues in the biosynthesis of Araf-containing complex carbohydrates. UDP-Araf itself is formed from UDP-arabinopyranose (UDP-Arap) by UDP-arabinopyranose mutase (UAM). However, the mechanism by which this enzyme catalyzes the interconversion of UDP-Arap and UDP-Araf has not been determined. To gain insight into this reaction, functionally recombinant rUAMs were reacted with UDP-Glc or UDP-Araf. The glycosylated recombinant UAMs were fragmented with trypsin, and the glycopeptides formed were then identified and sequenced by LC-MS/MS. The results of these experiments, together with site-directed mutagenesis studies, suggest that in functional UAMs an arginyl residue is reversibly glycosylated with a single glycosyl residue, and that this residue is required for mutase activity. We also provide evidence that a DXD motif is required for catalytic activity.     

Incorporation and remodeling of phosphatidylethanolamine containing short acyl residues in yeast

Phosphatidylethanolamine (PE) is one of the essential phospholipids in the yeast Saccharomyces cerevisiae. We have previously shown that a yeast strain, the endogenous PE synthesis of which was controllable, grew in the presence of PE containing decanoyl residues (diC10PE) when PE synthesis was repressed. In this study, we investigated the fate of diC10PE, its uptake and remodeling in yeast. Deletion of the genes encoding Lem3p/Ros3p or P-type ATPases, Dnf1p and Dnf2p, impaired the growth of the mutants in the medium containing diC10PE, suggesting the involvement of these proteins in the uptake of diC10PE. Analysis of the metabolism of deuterium-labeled diC10PE by electrospray ionization tandem mass spectrometry revealed that it was rapidly converted to deuterium-labeled PEs containing C16 or C18 acyl residues. The probable intermediate PEs that contained decanoic acid and C16 or C18 fatty acids as acyl residues were also detected. In addition, a substantial amount of decanoic acid was released into the culture medium during growth in the presence of diC10PE. These results imply that diC10PE was remodeled to PEs with longer acyl residues and used as membrane components. Defects in the remodeling of diC10PE in the deletion mutants of ALE1 and SLC1, products of which were capable of acyl-transfer to the sn− 2 position of lyso-phospholipids, suggested their involvement in the introduction of acyl residues to the sn− 2 position of lyso-phosphatidylethanolamine in the remodeling reaction of diC10PE. Our results also suggest the presence of a mechanism to maintain the physiological length of PE acyl residues in yeast.     

RNA模式分析进展

研究表明,R N A模式在基因表达调控方面起着重要作用.由于RN A模式不仅与初级序列有关,更多的表现为高级结构(一般为二级结构)的保守性,所以R N A模式的识别比D N A模式的识别要复杂的多.近十几年里,对R N A模式分析作了大量的计算方面的研究,包括:R N A结构的预测、识别和已知的类型相似的R N A模式、在一组功能或进化相关的基因中找出共同的R N A模式.这里对上述3个方面的计算方法的发展和研究进行了综述.