Turning points in the evolution of peroxidase–catalase superfamily: molecular phylogeny of hybrid heme peroxidases

Heme peroxidases and catalases are key enzymes of hydrogen peroxide metabolism and signaling. Here, the reconstruction of the molecular evolution of the peroxidase–catalase superfamily (annotated in pfam as PF00141) based on experimentally verified as well as numerous newly available genomic sequences is presented. The robust phylogenetic tree of this large enzyme superfamily was obtained from 490 full-length protein sequences. Besides already well-known families of heme b peroxidases arranged in three main structural classes, completely new (hybrid type) peroxidase families are described being located at the border of these classes as well as forming (so far missing) links between them. Hybrid-type A peroxidases represent a minor eukaryotic subfamily from Excavates, Stramenopiles and Rhizaria sharing enzymatic and structural features of ascorbate and cytochrome c peroxidases. Hybrid-type B peroxidases are shown to be spread exclusively among various fungi and evolved in parallel with peroxidases in land plants. In some ascomycetous hybrid-type B peroxidases, the peroxidase domain is fused to a carbohydrate binding (WSC) domain. Both here described hybrid-type peroxidase families represent important turning points in the complex evolution of the whole peroxidase–catalase superfamily. We present and discuss their phylogeny, sequence signatures and putative biological function.     

New record and southern range extension for the Mearn's grasshopper mouse ( Onychomys arenicola Mearns, 1896) in Veracruz, Mexico

Mearn’s grasshopper mouse ( Onychomys arenicola ) in Mexico is found primarily in the central and northern states. This is the first report of the genus Onychomys in the state of Veracruz, based on 7 captured specimens (3 collected). This finding extends the species’ known distribution by approximately 470 km east-southeast. In addition, this record increases the alpha diversity of terrestrial mammals for the state of Veracruz to 192 species.     

Participation of the second extracellular loop of claudin-5 in paracellular tightening against ions, small and large molecules

Tight junctions control paracellular permeability. Here, we analyzed the impact of residues in the second extracellular loop (ECL2) of mouse claudin-5 on paracellular permeability. Stable expression of claudin-5_{wild type} in MDCK-II cells—but not that of mutants R145A, Y148A, Y158A or E159Q—increased transepithelial electrical resistance and decreased fluorescein permeation. Expression of claudin-5_Y148A, _Y158A or _E159Q enhanced permeability of FITC-dextran_10 kDa, which was unchanged in cells expressing claudin-5_{wild type} or claudin-5_R145A. In contrast, targeting to tight junctions, strand morphology and tight junction assembly were unchanged. It is concluded that R145 is unessential for trans-interaction of claudin-5, but necessary for tightening against small solutes and ions. The highly conserved residues Y148, Y158 and E159 in ECL2 of claudin-5 contribute to homo- and/or heterophilic trans-interaction between classic claudins and thereby tighten the paracellular space against ions, small and large molecules. These results provide novel insights into the molecular function of tight junctions.     

Forecasting Private Consumption by Consumer Surveys

Survey‐based indicators are widely seen as leading indicators for economic activity. As such, consumer confidence might be informative for the future path of private consumption. Although the indicators receive high attention in the media, their forecasting power often appears to be very limited. This paper takes a fresh look at the data that serve as a basis for the consumer confidence indicator (CCI) reported by the EU Commission for the euro area. Different pooling methods are applied to exploit the survey information. Forecasts are based on mixed data sampling (MIDAS) and bridge equations. While the CCI does not outperform the autoregressive benchmark, the new indicators are able to raise forecasting performance. The best performing indicator should be built upon pre‐selection methods. Data‐driven aggregation methods should be preferred to determine the weights of the individual ingredients. Copyright © 2011 John Wiley & Sons, Ltd.     

Replacement of huntingtin exon 1 by trans-splicing

Huntington’s disease (HD) is an autosomal-dominant neurodegenerative disorder caused by polyglutamine expansion in the amino-terminus of huntingtin (HTT). HD offers unique opportunities for promising RNA-based therapeutic approaches aimed at reducing mutant HTT expression, since the HD mutation is considered to be a “gain-of-function” mutation. Allele-specific strategies that preserve expression from the wild-type allele and reduce the levels of mutant protein would be of particular interest. Here, we have conducted proof-of-concept studies to demonstrate that spliceosome-mediated trans-splicing is a viable molecular strategy to specifically repair the HTT allele. We employed a dual plasmid transfection system consisting of a pre-mRNA trans-splicing module (PTM) containing HTT exon 1 and a HTT minigene to demonstrate that HTT exon 1 can be replaced in trans. We detected the presence of the trans-spliced RNA in which PTM exon 1 was correctly joined to minigene exons 2 and 3. Furthermore, exon 1 from the PTM was trans-spliced to the endogenous HTT pre-mRNA in cultured cells as well as disease-relevant models, including HD patient fibroblasts and primary neurons from a previously described HD mouse model. These results suggest that the repeat expansion of HTT can be repaired successfully not only in the context of synthetic minigenes but also within the context of HD neurons. Therefore, pre-mRNA trans-splicing may be a promising approach for the treatment of HD and other dominant genetic disorders.     

Examining Conditions that Influence Evaluation use within a Humanitarian Non-Governmental Organization in Burkina Faso (West Africa)

Program evaluation can support capacity building and inform practice and policy. Yet long-term efforts to ensure evaluation use (EU) in the humanitarian sector are seldom documented, leaving much uncertainty about EU conditions. This study examined conditions that influenced EU by stakeholders of a humanitarian non-governmental organization (NGO) in Burkina Faso striving to base its health care program on solid evidence. It used 36 qualitative semi-structured interviews and a single case study design to document stakeholders’ (n?=?26) perception of EU conditions. Analyses focussed on characteristics of five broad conditions of research use previously documented. Results demonstrate that EU was facilitated by intended users with proactive attitudes, research experience, and willingness to participate in program evaluations. Also helpful was an organizational culture that valued learning, feedback, and accountability, wherein leaders collaborated toward common goals. Evaluation-based knowledge that met information needs and that was actionable, contextualized, and quickly accessible enhanced EU. Knowledge transfer strategies promoting EU were diverse, participatory, adapted to needs, and regularly followed up. Evaluators who were trusted, experienced, credible, and adaptable, promoted EU most effectively. Conversely, EU was compromised when intended users felt distrusting, uninformed, or unable to engage in program evaluations. Knowledge contradicting expectations or deemed inapplicable impeded EU. Adapting knowledge transfer strategies required time and interactions. Initially, evaluations were not sufficiently adapted and put into plain language, which hampered EU. EU conditions are numerous and intricately interrelated, but interpersonal relationships, trust, and effective communication are key conditions for evaluators and stakeholders wishing to promote EU.

     

多笔画联机手写字符识别的动态时间规正算法

在单笔画符号(或字符)联机手写识别中,动态时间规正(DTW)算法遵循时间次序约束和边界约束,并具有较高的识别率.为了将此算法应用于多笔画符号识别,常用而简单的方法是按照人们的手写顺序连接多笔画符号为单笔画符号.但此方法存在一个问题:人们常使用不同的笔画顺序和笔画方向书写同一个符号,用朴素(Brute Force)方法寻找所有笔画可能性非常耗时.为了降低计算复杂度,文中提出了DTW A*算法.在部分笔画匹配时,此算法保留着次序约束,并用A*算法降低计算复杂度.文中还通过流程图数据库多笔画符号识别实验对比了DTW A*算法、DTW算法、改良Hausdorff距离3种算法的性能,结果表明DTW A*算法具有最高的识别率和最好的稳定性.