Comparing de novo and reference-based transcriptome assembly strategies by applying them to the blood-sucking bug Rhodnius prolixus

High Throughput Sequencing capabilities have made the process of assembling a transcriptome easier, whether or not there is a reference genome. But the quality of a transcriptome assembly must be good enough to capture the most comprehensive catalog of transcripts and their variations, and to carry out further experiments on transcriptomics. There is currently no consensus on which of the many sequencing technologies and assembly tools are the most effective. Many non-model organisms lack a reference genome to guide the transcriptome assembly. One question, therefore, is whether or not a reference-based genome assembly gives better results than de novo assembly. The blood-sucking insect Rhodnius prolixus-a vector for Chagas disease-has a reference genome. It is therefore a good model on which to compare reference-based and de novo transcriptome assemblies. In this study, we compared de novo and reference-based genome assembly strategies using three datasets (454, Illumina, 454 combined with Illumina) and various assembly software. We developed criteria to compare the resulting assemblies: the size distribution and number of transcripts, the proportion of potentially chimeric transcripts, how complete the assembly was (completeness evaluated both through CEGMA software and R. prolixus proteome fraction retrieved). Moreover, we looked for the presence of two chemosensory gene families (Odorant-Binding Proteins and Chemosensory Proteins) to validate the assembly quality. The reference-based assemblies after genome annotation were clearly better than those generated using de novo strategies alone. Reference-based strategies revealed new transcripts, including new isoforms unpredicted by automatic genome annotation. However, a combination of both de novo and reference-based strategies gave the best result, and allowed us to assemble fragmented transcripts.

Développement de méthodes d'assemblage de génomes de novo adaptées aux bactéries endosymbiotes

Le but de ce projet était de développer des méthodes d'assemblage de novo dans le but d'assembler de petits génomes, principalement bactériens, à partir de données de séquençage de nouvelle-génération. Éventuellement, ces méthodes pourraient être appliquées à l'assemblage du génome de StachEndo, une Alpha-Protéobactérie inconnue endosymbiote de l'amibe Stachyamoeba lipophora. Suite à plusieurs analyses préliminaires, il fut observé que l’utilisation de lectures Illumina avec des assembleurs par graphe DeBruijn produisait les meilleurs résultats. Ces expériences ont également montré que les contigs produits à partir de différentes tailles de k-mères étaient complémentaires pour la finition des génomes. L’ajout de longues paires de lectures chevauchantes se montra essentiel pour la finition complète des grandes répétitions génomiques. Ces méthodes permirent d'assembler le génome de StachEndo (1,7 Mb). L'annotation de ce génome permis de montrer que StachEndo possède plusieurs caractéristiques inhabituelles chez les endosymbiotes. StachEndo constitue une espèce d'intérêt pour l'étude du développement endosymbiotique.

Achieving low cost and high quality aero structure assembly through integrated digital metrology systems

Measurement assisted assembly (MAA) has the potential to facilitate a step change in assembly efficiency for large structures such as airframes through the reduction of rework, manually intensive processes and expensive monolithic assembly tooling. It is shown how MAA can enable rapid part-to-part assembly, increased use of flexible automation, traceable quality assurance and control, reduced structure weight and improved aerodynamic tolerances. These advances will require the development of automated networks of measurement instruments; model based thermal compensation, the automatic integration of 'live' measurement data into variation simulation and algorithms to generate cutting paths for predictive shimming and drilling processes. This paper sets out an architecture for digital systems which will enable this integrated approach to variation management. © 2013 The Authors.

Factors Controlling The Deposition Of Silk Fibroin Nanofibrils During Layer-by-layer Assembly.

The layer-by-layer technique has been used as a powerful method to produce multilayer thin films with tunable properties. When natural polymers are employed, complicated phenomena such as self-aggregation and fibrilogenesis can occur, making it more difficult to obtain and characterize high-quality films. The weak acid and base character of such materials provides multilayer systems that may differ from those found with synthetic polymers due to strong self-organization effects. Specifically, LbL films prepared with chitosan and silk fibroin (SF) often involve the deposition of fibroin fibrils, which can influence the assembly process, surface properties, and overall film functionality. In this case, one has the intriguing possibility of realizing multilayer thin films with aligned nanofibers. In this article, we propose a strategy to control fibroin fibril formation by adjusting the assembly partner. Aligned fibroin fibrils were formed when chitosan was used as the counterpart, whereas no fibrils were observed when poly(allylamine hydrochloride) (PAH) was used. Charge density, which is higher in PAH, apparently stabilizes SF aggregates on the nanometer scale, thereby preventing their organization into fibrils. The drying step between the deposition of each layer was also crucial for film formation, as it stabilizes the SF molecules. Preliminary cell studies with optimized multilayers indicated that cell viability of NIH-3T3 fibroblasts remained between 90 and 100% after surface seeding, showing the potential application of the films in the biomedical field, as coatings and functional surfaces.

Ergonomic Planned Supply in an Automotive Assembly Line

The objective of this research is to identify the benefits of ergonomic improvements in workstations and in planned parts supply in an automotive assembly line. Another aim is to verify to what extent it is possible to create competitive advantages in the manufacturing area with reduction in vehicle assembly time by using technological investments in ergonomics with benefits to the worker and to the company. The Methods Time Measurement (MTM) methodology is chosen to measure the process time differences. To ensure a reliable comparison, a company in Brazil that has two different types of assembly line installations in the same plant was observed, and both assembly lines were under the same influences in terms of human resources, wages, food, and educational level of the staff. In this article, the first assembly line is called ""new"" and was built 6 years ago, with high investments in ergonomic solutions, in the supply system, and in the process. The other is called ""traditional"" and was built 23 years ago with few investments in the area. (C) 2010 Wiley Periodicals, Inc.

Quantitative description of the self-assembly of patchy particles into chains and rings

We numerically study a simple fluid composed of particles having a hard-core repulsion complemented by two patchy attractive sites on the particle poles. An appropriate choice of the patch angular width allows for the formation of ring structures which, at low temperatures and low densities, compete with the growth of linear aggregates. The simplicity of the model makes it possible to compare simulation results and theoretical predictions based on the Wertheim perturbation theory, specialized to the case in which ring formation is allowed. Such a comparison offers a unique framework for establishing the quality of the analytic predictions. We find that the Wertheim theory describes remarkably well the simulation results.

Quality Assessment of Cardiac Magnetic Resonance Images at 1.5/3.0 T: Description and Validation of Standardized Criteria

PURPOSE: Cardiovascular magnetic resonance (CMR) has become a robust and important diagnostic imaging modality in cardiovascular medicine. However,insufficient image quality may compromise its diagnostic accuracy. No standardized criteria are available to assess the quality of CMR studies. We aimed todescribe and validate standardized criteria to evaluate the quality of CMR studies including: a) cine steady-state free precession, b) delayed gadoliniumenhancement, and c) adenosine stress first-pass perfusion. These criteria will serve for the assessment of the image quality in the setting of the Euro-CMR registry.METHOD AND MATERIALS: First, a total of 45 quality criteria were defined (35 qualitative criteria with a score from 0-3, and 10 quantitative criteria). Thequalitative score ranged from 0 to 105. The lower the qualitative score, the better the quality. The quantitative criteria were based on the absolute signal intensity (delayed enhancement) and on the signal increase (perfusion) of the anterior/posterior left ventricular wall after gadolinium injection. These criteria were then applied in 30 patients scanned with a 1.5T system and in 15 patients scanned with a 3.0T system. The examinations were jointly interpreted by 3 CMR experts and 1 study nurse. In these 45 patients the correlation between the results of the quality assessment obtained by the different readers was calculated.RESULTS: On the 1.5T machine, the mean quality score was 3.5. The mean difference between each pair of observers was 0.2 (5.7%) with a mean standarddeviation of 1.4. On the 3.0T machine, the mean quality score was 4.4. The mean difference between each pair of onservers was 0.3 (6.4%) with a meanstandard deviation of 1.6. The quantitative quality assessments between observers were well correlated for the 1.5T machine: R was between 0.78 and 0.99 (pCONCLUSION: The described criteria for the assessment of CMR image quality are robust and have a low inter-observer variability, especially on 1.5T systems.CLINICAL RELEVANCE/APPLICATION: These criteria will allow the standardization of CMR examinations. They will help to improve the overall quality ofexaminations and the comparison between clinical studies.

Visualization and quality assessment of de novo genome assemblies.

Recent technological progress has greatly facilitated de novo genome sequencing. However, de novo assemblies consist in many pieces of contiguous sequence (contigs) arranged in thousands of scaffolds instead of small numbers of chromosomes. Confirming and improving the quality of such assemblies is critical for subsequent analysis. We present a method to evaluate genome scaffolding by aligning independently obtained transcriptome sequences to the genome and visually summarizing the alignments using the Cytoscape software. Applying this method to the genome of the red fire ant Solenopsis invicta allowed us to identify inconsistencies in 7%, confirm contig order in 20% and extend 16% of scaffolds.Scripts that generate tables for visualization in Cytoscape from FASTA sequence and scaffolding information files are publicly available at https://github.com/ksanao/TGNet.

De novo transcriptome assembly and annotation of the isopod Idotea balthica

The Baltic Sea is a unique environment that contains unique genetic populations. In order to study these populations on a genetic level basic molecular research is needed. The aim of this thesis was to provide a basic genetic resource for population genomic studies by de novo assembling a transcriptome for the Baltic Sea isopod Idotea balthica. RNA was extracted from a whole single adult male isopod and sequenced using Illumina (125bp PE) RNA-Seq. The reads were preprocessed using FASTQC for quality control, TRIMMOMATIC for trimming, and RCORRECTOR for error correction. The preprocessed reads were then assembled with TRINITY, a de Bruijn graph-based assembler, using different k-mer sizes. The different assemblies were combined and clustered using CD-HIT. The assemblies were evaluated using TRANSRATE for quality and filtering, BUSCO for completeness, and TRANSDECODER for annotation potential. The 25-mer assembly was annotated using PANNZER (protein annotation with z-score) and BLASTX. The 25-mer assembly represents the best first draft assembly since it contains the most information. However, this assembly shows high levels of polymorphism, which currently cannot be differentiated as paralogs or allelic variants. Furthermore, this assembly is incomplete, which could be improved by sampling additional developmental stages.

A Study of Ordered Gene Problems Featuring DNA Error Correction and DNA Fragment Assembly with a Variety of Heuristics, Genetic Algorithm Variations, and Dynamic Representations

Ordered gene problems are a very common classification of optimization problems. Because of their popularity countless algorithms have been developed in an attempt to find high quality solutions to the problems. It is also common to see many different types of problems reduced to ordered gene style problems as there are many popular heuristics and metaheuristics for them due to their popularity. Multiple ordered gene problems are studied, namely, the travelling salesman problem, bin packing problem, and graph colouring problem. In addition, two bioinformatics problems not traditionally seen as ordered gene problems are studied: DNA error correction and DNA fragment assembly. These problems are studied with multiple variations and combinations of heuristics and metaheuristics with two distinct types or representations. The majority of the algorithms are built around the Recentering- Restarting Genetic Algorithm. The algorithm variations were successful on all problems studied, and particularly for the two bioinformatics problems. For DNA Error Correction multiple cases were found with 100% of the codes being corrected. The algorithm variations were also able to beat all other state-of-the-art DNA Fragment Assemblers on 13 out of 16 benchmark problem instances.

Re-Engineering Towed Arrays for Quality Enhancement - Network Based Towed Array

The Towed Array electronics is a multi-channel simultaneous real time high speed data acquisition system. Since its assembly is highly manpower intensive, the costs of arrays are prohibitive and therefore any attempt to reduce the manufacturing, assembly, testing and maintenance costs is a welcome proposition. The Network Based Towed Array is an innovative concept and its implementation has remarkably simplified the fabrication, assembly and testing and revolutionised the Towed Array scenario. The focus of this paper is to give a good insight into the Reliability aspects of Network Based Towed Array. A case study of the comparison between the conventional array and the network based towed array is also dealt with

Knowledge transfer of the Western concept of ‘quality’

This paper addresses one of the issues in contemporary globalisation theory: the extent to which there is ‘one best way’ in which business can be done and organisations managed. It uses Czarniawska’s ‘Travels of Ideas’ model as an organising framework to present and understand how the concept of ‘Quality’, so important in contemporary approaches to manufacturing & services, and their management, travelled to, and impinged on, a newly opened vehicle assembly plant in Poland. The extent to which new meanings were mutually created in the process of translation is discussed, using ethnographic reporting and analysis techniques commonly used in diffusion research. Parallels between the process of translation as an idea becomes embedded into a new cultural location, and the processes which contemporary research has identified as important to organisational learning, are briefly discussed in conclusion.

Studies of Design and Assembly Defects on Integrated and Modular Architectures

It is known that despite companies’ efforts to improve the quality of their products, design and assembly defects results in large repair costs both in terms of repair and providing feedback to the origin of the defect. The purpose of this paper is to study these types of defects and the defect rates in design and assembly. The paper presents a web based questionnaire answered by 29 companies. The result shows that the defect rate (defects per product) spanned from 0.01 to 10. Also, design and assembly defects covered 46%, 23% respectively, of all occurred defects. A case study is also presented, performed at a company who recently implemented a modular architecture. In this company, defects from 5 700 integrated product architectures are compared with defects from 431 modular architectures. The average defect rate increased by 21.5% – from 0.65 to 0.79 – when a more modular architecture has been implemented. Furthermore, the study showed that the assembly defects have decreased while the design defects increased. The results presented in this paper will also support the development of the MPV (Module Property Verification) method which is briefly described.