Development and evaluation of an automated annotation pipeline and cDNA annotation system

Manual curation has long been held to be the gold standard for functional annotation of DNA sequence. Our experience with the annotation of more than 20,000 full-length cDNA sequences revealed problems with this approach, including inaccurate and inconsistent assignment of gene names, as well as many good assignments that were difficult to reproduce using only computational methods. For the FANTOM2 annotation of more than 60,000 cDNA clones, we developed a number of methods and tools to circumvent some of these problems, including an automated annotation pipeline that provides high-quality preliminary annotation for each sequence by introducing an uninformative filter that eliminates uninformative annotations, controlled vocabularies to accurately reflect both the functional assignments and the evidence supporting them, and a highly refined, Web-based manual annotation tool that allows users to view a wide array of sequence analyses and to assign gene names and putative functions using a consistent nomenclature. The ultimate utility of our approach is reflected in the low rate of reassignment of automated assignments by manual curation. Based on these results, we propose a new standard for large-scale annotation, in which the initial automated annotations are manually investigated and then computational methods are iteratively modified and improved based on the results of manual curation.

Definition and spatial annotation of the dynamic secretome during early kidney development

The term secretome has been defined as a set of secreted proteins (Grimmond et al. [2003] Genome Res 13:1350-1359). The term secreted protein encompasses all proteins exported from the cell including growth factors, extracellular proteinases, morphogens, and extracellular matrix molecules. Defining the genes encoding secreted proteins that change in expression during organogenesis, the dynamic secretome, is likely to point to key drivers of morphogenesis. Such secreted proteins are involved in the reciprocal interactions between the ureteric bud (UB) and the metanephric mesenchyme (AM) that occur during organogenesis of the metanephros. Some key metanephric secreted proteins have been identified, but many remain to be determined. In this study, microarray expression profiling of E10.5, E11.5, and E13.5 kidney and consensus bioinformatic analysis were used to define a dynamic secretome of early metanephric development. In situ hybridisation was used to confirm microarray results and clarify spatial expression patterns for these genes. Forty-one secreted factors were dynamically expressed between the E10.5 and E13.5 timeframe profiled, and 25 of these factors had not previously been implicated in kidney development. A text-based anatomical ontology was used to spatially annotate the expression pattern of these genes in cultured metanephric explants.

The Wnt receptor Ryk is required for Wnt5a-mediated axon guidance on the contralateral side of the corpus callosum

Ryk (receptor related to tyrosine kinase) has been shown to be a novel Wnt receptor in both Caenorhabditis elegans and Drosophila melanogaster. Recently, Ryk-Wnt interactions were shown to guide corticospinal axons down the embryonic mouse spinal cord. Here we show that, in Ryk-deficient mice, cortical axons project aberrantly across the major forebrain commissure, the corpus callosum. Many mouse mutants have been described in which loss-of-function mutations result in the inability of callosal axons to cross the midline, thereby forming Probst bundles on the ipsilateral side. In contrast, loss of Ryk does not interfere with the ability of callosal axons to cross the midline but impedes their escape from the midline into the contralateral side. Therefore, Ryk(-/-) mice display a novel callosal guidance phenotype. We also show that Wnt5a acts as a chemorepulsive ligand for Ryk, driving callosal axons toward the contralateral hemisphere after crossing the midline. In addition, whereas callosal axons do cross the midline in Ryk(-/-) embryos, they are defasciculated on the ipsilateral side, indicating that Ryk also promotes fasciculation of axons before midline crossing. In summary, this study expands the emerging role for Wnts in axon guidance and identifies Ryk as a key guidance receptor in the establishment of the corpus callosum. Our analysis of Ryk function further advances our understanding of the molecular mechanisms underlying the formation of this important commissure.

Transcript annotation in FANTOM3: Mouse gene catalog based on physical cDNAs

T he international FANTOM consortium aims to produce a comprehensive picture of the mammalian transcriptome, based upon an extensive cDNA collection and functional annotation of full-length enriched cDNAs. The previous dataset, FANTOM(2), comprised 60,770 full- length enriched cDNAs. Functional annotation revealed that this cDNA dataset contained only about half of the estimated number of mouse protein- coding genes, indicating that a number of cDNAs still remained to be collected and identified. To pursue the complete gene catalog that covers all predicted mouse genes, cloning and sequencing of full- length enriched cDNAs has been continued since FANTOM2. In FANTOM3, 42,031 newly isolated cDNAs were subjected to functional annotation, and the annotation of 4,347 FANTOM2 cDNAs was updated. To accomplish accurate functional annotation, we improved our automated annotation pipeline by introducing new coding sequence prediction programs and developed a Web- based annotation interface for simplifying the annotation procedures to reduce manual annotation errors. Automated coding sequence and function prediction was followed with manual curation and review by expert curators. A total of 102,801 full- length enriched mouse cDNAs were annotated. Out of 102,801 transcripts, 56,722 were functionally annotated as protein coding ( including partial or truncated transcripts), providing to our knowledge the greatest current coverage of the mouse proteome by full- length cDNAs. The total number of distinct non- protein- coding transcripts increased to 34,030. The FANTOM3 annotation system, consisting of automated computational prediction, manual curation, and. nal expert curation, facilitated the comprehensive characterization of the mouse transcriptome, and could be applied to the transcriptomes of other species.

Development of a multi-tiered speech annotation system for accented English

This paper discusses methodological issues in the development of a multitiered, phonetic annotation system, intended to capture pronunciation variation in the speech of second language learners and to serve in construction of a data base for training ASR models to recognize major pronunciation variants in the assessment of accented English.

A corpus-based analysis of transfer effects and connected speech processes in Vietnamese English

This paper presents a corpus-based descriptive analysis of the most prevalent transfer effects and connected speech processes observed in a comparison of 11 Vietnamese English speakers (6 females, 5 males) and 12 Australian English speakers (6 males, 6 females) over 24 grammatical paraphrase items. The phonetic processes are segmentally labelled in terms of IPA diacritic features using the EMU speech database system with the aim of labelling departures from native-speaker pronunciation. An analysis of prosodic features was made using ToBI framework. The results show many phonetic and prosodic processes which make non-native speakers’ speech distinct from native ones. The corpusbased methodology of analysing foreign accent may have implications for the evaluation of non-native accent, accented speech recognition and computer assisted pronunciation- learning.

Image annotation with Photocopain

Photo annotation is a resource-intensive task, yet is increasingly essential as image archives and personal photo collections grow in size. There is an inherent con?ict in the process of describing and archiving personal experiences, because casual users are generally unwilling to expend large amounts of e?ort on creating the annotations which are required to organise their collections so that they can make best use of them. This paper describes the Photocopain system, a semi-automatic image annotation system which combines information about the context in which a photograph was captured with information from other readily available sources in order to generate outline annotations for that photograph that the user may further extend or amend.

The science and technology of corpus, and corpus for science and technology

Corpus Linguistics is a young discipline. The earliest work was done in the 1960s, but corpora only began to be widely used by lexicographers and linguists in the late 1980s, by language teachers in the late 1990s, and by language students only very recently. This course in corpus linguistics was held at the Departamento de Linguistica Aplicada, E.T.S.I. de Minas, Universidad Politecnica de Madrid from June 15-19 1998. About 45 teachers registered for the course. 30% had PhDs in linguistics, 20% in literature, and the rest were doctorandi or qualified English teachers. The course was designed to introduce the use of corpora and other computational resources in teaching and research, with special reference to scientific and technological discourse in English. Each participant had a computer networked with the lecturer’s machine, whose display could be projected onto a large screen. Application programs were loaded onto the central server, and telnet and a web browser were available. COBUILD gave us permission to access the 323 million word Bank of English corpus, Mike Scott allowed us to use his Wordsmith Tools software, and Tim Johns gave us a copy of his MicroConcord program.

The discourse of climate change: a corpus-based approach

Based on Goffman’s definition that frames are general ‘schemata of interpretation’ that people use to ‘locate, perceive, identify, and label’, other scholars have used the concept in a more specific way to analyze media coverage. Frames are used in the sense of organizing devices that allow journalists to select and emphasise topics, to decide ‘what matters’ (Gitlin 1980). Gamson and Modigliani (1989) consider frames as being embedded within ‘media packages’ that can be seen as ‘giving meaning’ to an issue. According to Entman (1993), framing comprises a combination of different activities such as: problem definition, causal interpretation, moral evaluation, and/or treatment recommendation for the item described. Previous research has analysed climate change with the purpose of testing Downs’s model of the issue attention cycle (Trumbo 1996), to uncover media biases in the US press (Boykoff and Boykoff 2004), to highlight differences between nations (Brossard et al. 2004; Grundmann 2007) or to analyze cultural reconstructions of scientific knowledge (Carvalho and Burgess 2005). In this paper we shall present data from a corpus linguistics-based approach. We will be drawing on results of a pilot study conducted in Spring 2008 based on the Nexis news media archive. Based on comparative data from the US, the UK, France and Germany, we aim to show how the climate change issue has been framed differently in these countries and how this framing indicates differences in national climate change policies.

A new venture in corpus-based lexicography: towards a dictionary of academic English

This paper asserts the increasing importance of academic English in an increasingly Anglophone world, and looks at the differences between academic English and general English, especially in terms of vocabulary. The creation of wordlists has played an important role in trying to establish the academic English lexicon, but these wordlists are not based on appropriate data, or are implemented inappropriately. There is as yet no adequate dictionary of academic English, and this paper reports on new efforts at Aston University to create a suitable corpus on which such a dictionary could be based.