SNPmplexViewer--toward a cost-effective traceability system

<p>Abstract</p> <p>Background</p> <p>Beef traceability has become mandatory in many regions of the world and is typically achieved through the use of unique numerical codes on ear tags and animal passports. DNA-based traceability uses the animal's own DNA code to identify it and the products derived from it. Using <it>SNaPshot</it>, a primer-extension-based method, a multiplex of 25 SNPs in a single reaction has been practiced for reducing the expense of genotyping a panel of SNPs useful for identity control.</p> <p>Findings</p> <p>To further decrease <it>SNaPshot</it>'s cost, we introduced the Perl script <it>SNPmplexViewer</it>, which facilitates the analysis of trace files for reactions performed without the use of fluorescent size standards. <it>SNPmplexViewer </it>automatically aligns reference and target trace electropherograms, run with and without fluorescent size standards, respectively. <it>SNPmplexViewer </it>produces a modified target trace file containing a normalised trace in which the reference size standards are embedded. <it>SNPmplexViewer </it>also outputs aligned images of the two electropherograms together with a difference profile.</p> <p>Conclusions</p> <p>Modified trace files generated by <it>SNPmplexViewer </it>enable genotyping of <it>SnaPshot </it>reactions performed without fluorescent size standards, using common fragment-sizing software packages. <it>SNPmplexViewer</it>'s normalised output may also improve the genotyping software's performance. Thus, <it>SNPmplexViewer </it>is a general free tool enabling the reduction of <it>SNaPshot</it>'s cost as well as the fast viewing and comparing of trace electropherograms for fragment analysis. <it>SNPmplexViewer </it>is available at <url>http://cowry.agri.huji.ac.il/cgi-bin/SNPmplexViewer.cgi</url>.</p

DNA BARCODING OF FISH SPECIES FROM THE MEDITERRANEAN COAST OF ISRAEL

Accurately-classified genomic data in the Barcode of Life Data System (BOLD) database is vital to the protection and conservation of marine biodiversity in the Mediterranean Sea. The taxonomic classifications of 468 fish of 50 Mediterranean species were analyzed using the BOLD Identifier tool for variation in the cytochrome oxidase subunit I (COI) mitochondrial gene. Within species, nucleotide maximum composite likelihood was low with a mean of 0.0044±0.0008. Three presumptive species had significantly higher values e.g., Arnoglossus spp. (0.07), Torquigener flavimaculosus (0.013) and Boops boops (0.028). However, samples of Arnoglossus species were sub-classified into two groups that were finally identified as two different species e.g., Arnoglossus laterna and Arnoglossus thori. For the different species, BLAST searches against the BOLD database using our DNA barcoding data as the query sequences designated the most similar targets into groups. For each analyzed species, the similarity of the first and second threshold groups ranged from 95 to 99% and from 83 to 98%, respectively. Sequence based classification for the first threshold group was concordant with morphology-based identification. However, for 34 analyzed species (68%) overlaps of species between the two threshold groups hampered classification. Tree-based phylogeny analysis detected more than one cluster in the first threshold group for 22 out of 50 species, representing genetic subgroups and geographic origins. There was a tendency for higher conservation and lower number of clusters in the Lessepsian (Red Sea) migrant versus indigenous species

Scanning Mechanical Microscopy of Laser Ablated Volumes Related to Inductively Coupled Plasma-Mass Spectrometry

Scanning mechanical microscopy based on the point by point sampling of the target surface was used to characterize volumes of minerals ablated by laser pulses (Nd: YAG, = 1064 nm, 140 μs pulse-width). Differentiated volumes resulting from vaporization and exfoliation mechanisms were selectively measured. Ablated volumes of natural pyrite (cubic FeS2), marcasite (orthorhombic FeS2) and arsenopyrite AsFeS, were transported into an inductively coupled plasma torch for subsequent mass analysis. The log of the S34 Fe57, and As75 mass intensities was linearly correlated with the log of the dimensions of the vaporized crater induced by the laser shots while large particles had no effect on the measured intensities. A memory effect for As was observed when a nylon tube was used to carry the ablated materials into the plasma torch. The memory effect was decreased by using a copper tube resulting probably from a difference in the electrical properties of the tubing systems leading to a lower adsorption of As within the copper tube than for the case of the nylon tube

Keeping the Past Alive: Memories of Israeli Jews at the Turn of the Millennium

We asked in an open-ended way in 1999–2000 what national and world events Israeli Jews consider most important from the past 60 years. Ten events were identified as foremost, including three from the time of independence and one that was quite recent. All the major memories are associated with efforts of the state through commemorations and in other ways to create a unitary collective memory. Five social background variables help account for which events are mentioned as most important: birth cohort, education, gender, ethnic origin, and religiosity. Other specific factors such as personal Holocaust experience and voting preferences are also considered.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/45656/1/11206_2004_Article_460743.pd

Coupling computer-interpretable guidelines with a drug-database through a web-based system – The PRESGUID project

BACKGROUND: Clinical Practice Guidelines (CPGs) available today are not extensively used due to lack of proper integration into clinical settings, knowledge-related information resources, and lack of decision support at the point of care in a particular clinical context. OBJECTIVE: The PRESGUID project (PREScription and GUIDelines) aims to improve the assistance provided by guidelines. The project proposes an online service enabling physicians to consult computerized CPGs linked to drug databases for easier integration into the healthcare process. METHODS: Computable CPGs are structured as decision trees and coded in XML format. Recommendations related to drug classes are tagged with ATC codes. We use a mapping module to enhance computerized guidelines coupling with a drug database, which contains detailed information about each usable specific medication. In this way, therapeutic recommendations are backed up with current and up-to-date information from the database. RESULTS: Two authoritative CPGs, originally diffused as static textual documents, have been implemented to validate the computerization process and to illustrate the usefulness of the resulting automated CPGs and their coupling with a drug database. We discuss the advantages of this approach for practitioners and the implications for both guideline developers and drug database providers. Other CPGs will be implemented and evaluated in real conditions by clinicians working in different health institutions

Implementation and performance of adaptive mesh refinement in the Ice Sheet System Model (ISSM v4.14)

Accurate projections of the evolution of ice sheets in a changing climate require a fine mesh/grid resolution in ice sheet models to correctly capture fundamental physical processes, such as the evolution of the grounding line, the region where grounded ice starts to float. The evolution of the grounding line indeed plays a major role in ice sheet dynamics, as it is a fundamental control on marine ice sheet stability. Numerical modeling of a grounding line requires significant computational resources since the accuracy of its position depends on grid or mesh resolution. A technique that improves accuracy with reduced computational cost is the adaptive mesh refinement (AMR) approach. We present here the implementation of the AMR technique in the finite element Ice Sheet System Model (ISSM) to simulate grounding line dynamics under two different benchmarks: MISMIP3d and MISMIP+. We test different refinement criteria: (a) distance around the grounding line, (b) a posteriori error estimator, the Zienkiewicz–Zhu (ZZ) error estimator, and (c) different combinations of (a) and (b). In both benchmarks, the ZZ error estimator presents high values around the grounding line. In the MISMIP+ setup, this estimator also presents high values in the grounded part of the ice sheet, following the complex shape of the bedrock geometry. The ZZ estimator helps guide the refinement procedure such that AMR performance is improved. Our results show that computational time with AMR depends on the required accuracy, but in all cases, it is significantly shorter than for uniformly refined meshes. We conclude that AMR without an associated error estimator should be avoided, especially for real glaciers that have a complex bed geometry.</p

Decoding of Superimposed Traces Produced by Direct Sequencing of Heterozygous Indels

Direct Sanger sequencing of a diploid template containing a heterozygous insertion or deletion results in a difficult-to-interpret mixed trace formed by two allelic traces superimposed onto each other. Existing computational methods for deconvolution of such traces require knowledge of a reference sequence or the availability of both direct and reverse mixed sequences of the same template. We describe a simple yet accurate method, which uses dynamic programming optimization to predict superimposed allelic sequences solely from a string of letters representing peaks within an individual mixed trace. We used the method to decode 104 human traces (mean length 294 bp) containing heterozygous indels 5 to 30 bp with a mean of 99.1% bases per allelic sequence reconstructed correctly and unambiguously. Simulations with artificial sequences have demonstrated that the method yields accurate reconstructions when (1) the allelic sequences forming the mixed trace are sufficiently similar, (2) the analyzed fragment is significantly longer than the indel, and (3) multiple indels, if present, are well-spaced. Because these conditions occur in most encountered DNA sequences, the method is widely applicable. It is available as a free Web application Indelligent at http://ctap.inhs.uiuc.edu/dmitriev/indel.asp

Anti-CRISPR AcrIIA5 Potently Inhibits All Cas9 Homologs Used for Genome Editing

CRISPR-Cas9 systems provide powerful tools for genome editing. However, optimal employment of this technology will require control of Cas9 activity so that the timing, tissue specificity, and accuracy of editing may be precisely modulated. Anti-CRISPR proteins, which are small, naturally occurring inhibitors of CRISPR-Cas systems, are well suited for this purpose. A number of anti-CRISPR proteins have been shown to potently inhibit subgroups of CRISPR-Cas9 systems, but their maximal inhibitory activity is generally restricted to specific Cas9 homologs. Since Cas9 homologs vary in important properties, differing Cas9s may be optimal for particular genome-editing applications. To facilitate the practical exploitation of multiple Cas9 homologs, here we identify one anti-CRISPR, called AcrIIA5, that potently inhibits nine diverse type II-A and type II-C Cas9 homologs, including those currently used for genome editing. We show that the activity of AcrIIA5 results in partial in vivo cleavage of a single-guide RNA (sgRNA), suggesting that its mechanism involves RNA interaction

Diverse M-Best Solutions by Dynamic Programming

Many computer vision pipelines involve dynamic programming primitives such as finding a shortest path or the minimum energy solution in a tree-shaped probabilistic graphical model. In such cases, extracting not merely the best, but the set of M-best solutions is useful to generate a rich collection of candidate proposals that can be used in downstream processing. In this work, we show how M-best solutions of tree-shaped graphical models can be obtained by dynamic programming on a special graph with M layers. The proposed multi-layer concept is optimal for searching M-best solutions, and so flexible that it can also approximate M-best diverse solutions. We illustrate the usefulness with applications to object detection, panorama stitching and centerline extraction