33 research outputs found

    Convergence of Macroscopic Tongue Anatomy in Ruminants and Scaling Relationships with Body Mass or Tongue Length

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    Various morphological measures demonstrate convergent evolution in ruminants with their natural diet, in particular with respect to the browser/grazer dichotomy. Here, we report quantitative macroanatomical measures of the tongue (length and width of specific parts) of 65 ruminant species and relate them to either body mass (BM) or total tongue length, and to the percentage of grass in the natural diet (%grass). Models without and with accounting for the phylogenetic structures of the dataset were used, and models were ranked using Akaike’s Information Criterion. Scaling relationships followed geometric principles, that is, length measures scaled with BM to the power of 0.33. Models that used tongue length rather than BM as a body size proxy were consistently ranked better, indicating that using size proxies that are less susceptible to a wider variety of factors (such as BM that fluctuates with body condition) should be attempted whenever possible. The proportion of the freely mobile tongue tip of the total tongue (and hence also the corpus length) was negatively correlated to %grass, in accordance with concepts that the feeding mechanism of browsers requires more mobile tongues. It should be noted that some nonbrowsers, such as cattle, use a peculiar mechanism for grazing that also requires long, mobile tongues, but they appear to be exceptions. A larger corpus width with increasing %grass corresponds to differences in snout shape with broader snouts in grazers. The Torus linguae is longer with increasing %grass, a finding that still warrants functional interpretation. This study shows that tongue measures covary with diet in ruminants. In contrast, the shape of the tongue (straight or “hourglassshaped” as measured by the ratio of the widest and smallest corpus width) is unrelated to diet and is influenced strongly by phylogeny

    Multi-Site N-glycan mapping study 1: Capillary electrophoresis – laser induced fluorescence

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    An international team that included 20 independent laboratories from biopharmaceutical companies, universities, analytical contract laboratories and national authorities in the United States, Europe and Asia was formed to evaluate the reproducibility of sample preparation and analysis of N-glycans using capillary electrophoresis of 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled glycans with laser induced fluorescence (CE-LIF) detection (16 sites) and ultra highperformance liquid chromatography (UHPLC, 12 sites; results to be reported in a subsequent publication). All participants used the same lot of chemicals, samples, reagents, and columns/capillaries to run their assays. Migration time, peak area and peak area percent values were determined for all peaks with >0.1% peak area. Our results demonstrated low variability and high reproducibility, both, within any given site as well across all sites, which indicates that a standard N-glycan analysis platform appropriate for general use (clone selection, process development, lot release, etc.) within the industry can be established

    Habitat quality, configuration and context effects on roe deer fecundity across a forested landscape mosaic

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    Effective landscape-scale management of source-sink deer populations will be strengthened by understanding whether local variation in habitat quality drives heterogeneity in productivity. We related female roe deer Capreolus capreolus fecundity and body mass to habitat composition and landscape context, separately for adults and yearlings, using multi-model inference (MMI) applied to a large sample of individuals (yearlings: fecundity=202, body mass=395; adults: fecundity=908, body mass=1669) culled during 2002-2015 from an extensive (195 km2) heterogeneous forest landscape. Adults were heavier (inter-quartile, IQ, effect size=+0.5kg) when culled in buffers comprising more arable lands while contrary to our prediction no effects on body mass of grassland, young forest or access to vegetation on calcareous soil were found. Heavier adults were more fertile (IQ effect size, +12% probability of having two embryos instead of one or zero). Counter-intuitively, adults with greater access to arable lands were less fecund (IQ effect of arable: -7% probability of having two embryos, instead of one or zero), and even accounting for greater body mass of adults with access to arable, their modelled fecundity was similar to or lower than that of adults in the forest interior. In contrast, effects of grassland, young forest and calcareous soil did not receive support. Yearling body mass had an effect on fecundity twice that found in adults (+23% probability of having one additional embryo), but yearling body mass and fecundity were not affected by any candidate habitat or landscape variables. Effect of arable lands on body mass and fecundity were small, with little variance explained (Coefficient of Variation of predicted fecundity across forest sub-regions=0.03 for adults). More variance in fecundity was attributed to other differences between forest management sub-regions (modelled as random effects), suggesting other factors might be important. When analysing source-sink population dynamics to support management, an average value of fecundity can be appropriate across a heterogeneous forest landscape

    Microarray Profiling of Phage-Display Selections for Rapid Mapping of Transcription Factor–DNA Interactions

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    Modern computational methods are revealing putative transcription-factor (TF) binding sites at an extraordinary rate. However, the major challenge in studying transcriptional networks is to map these regulatory element predictions to the protein transcription factors that bind them. We have developed a microarray-based profiling of phage-display selection (MaPS) strategy that allows rapid and global survey of an organism's proteome for sequence-specific interactions with such putative DNA regulatory elements. Application to a variety of known yeast TF binding sites successfully identified the cognate TF from the background of a complex whole-proteome library. These factors contain DNA-binding domains from diverse families, including Myb, TEA, MADS box, and C2H2 zinc-finger. Using MaPS, we identified Dot6 as a trans-active partner of the long-predicted orphan yeast element Polymerase A & C (PAC). MaPS technology should enable rapid and proteome-scale study of bi-molecular interactions within transcriptional networks

    Genetic variation and exercise-induced muscle damage: implications for athletic performance, injury and ageing.

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    Prolonged unaccustomed exercise involving muscle lengthening (eccentric) actions can result in ultrastructural muscle disruption, impaired excitation-contraction coupling, inflammation and muscle protein degradation. This process is associated with delayed onset muscle soreness and is referred to as exercise-induced muscle damage. Although a certain amount of muscle damage may be necessary for adaptation to occur, excessive damage or inadequate recovery from exercise-induced muscle damage can increase injury risk, particularly in older individuals, who experience more damage and require longer to recover from muscle damaging exercise than younger adults. Furthermore, it is apparent that inter-individual variation exists in the response to exercise-induced muscle damage, and there is evidence that genetic variability may play a key role. Although this area of research is in its infancy, certain gene variations, or polymorphisms have been associated with exercise-induced muscle damage (i.e. individuals with certain genotypes experience greater muscle damage, and require longer recovery, following strenuous exercise). These polymorphisms include ACTN3 (R577X, rs1815739), TNF (-308 G>A, rs1800629), IL6 (-174 G>C, rs1800795), and IGF2 (ApaI, 17200 G>A, rs680). Knowing how someone is likely to respond to a particular type of exercise could help coaches/practitioners individualise the exercise training of their athletes/patients, thus maximising recovery and adaptation, while reducing overload-associated injury risk. The purpose of this review is to provide a critical analysis of the literature concerning gene polymorphisms associated with exercise-induced muscle damage, both in young and older individuals, and to highlight the potential mechanisms underpinning these associations, thus providing a better understanding of exercise-induced muscle damage

    MaPS identifies <i>RAP1</i> as the gene whose product interacts with the Rap1 binding site.

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    <p>The yeast genomic DNA phage display library was selected for three rounds against a double-stranded oligonucleotide and PCR products of an upstream region containing Rap1 binding sites. The selected population of phage were profiled through microarray hybridization. Displayed is the distribution of the mean percentile rank for five independent such selections performed. The ORF corresponding to Rap1 had the highest mean percentile rank out of a total of 6242 ORFs queried on the array.</p

    The Dot6 protein is a sequence-specific PAC element binding factor.

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    <p>Gel shift assay was performed with Dot6 DNA-binding domain and DNA containing PAC elements. Purified recombinant GST-Dot6 was incubated with 50 fM biotinylated probe containing 4 copies of the PAC element (bPAC4). Unbiotinylated competitor (PAC4) had identical sequence to the probe, or mutations in each copy of the PAC element (XPAC4).</p

    The Rap1 DNA-binding domain is enriched in a sequence-specific and salt-dependent manner.

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    <p>A phage display library was affinity selected against indicated quantities of double-stranded DNA containing Rap1 binding sites under indicated salt conditions. Results from PCR of input phage library (input) or after second round of selection are shown. The intensity of a band is proportional to its abundance in the library. Lanes designated RAP1 indicate results of specific PCR against a single phage with the <i>RAP1</i> DNA-binding domain known to be in the library and the red arrows mark the expected size of this clone. Gel-isolation and sequencing of the selected bands at this location confirmed that they correspond to this clone. The blue arrows point out the PCR products corresponding to the <i>MCM1</i> DNA-binding domain (lower band) and <i>MCM1</i> ORF (upper band). The remaining bands show variable enrichment as a function of salt concentration and likely represent non-specific enrichment during the selection.</p

    An overview of Microarray profiling of phage-display selection technology.

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    <p>(A) 1–3 kb fragments of yeast genomic DNA are cloned into T7 bacteriophage to create a translational fusion between the capsid protein and the peptide sequence encoded by the insert. (B) The library of phage are exposed to immobilized target DNA molecules and non-binding phage are washed away. Bound phage are eluted, amplified in liquid culture, and the process is repeated over multiple rounds. The sequence content of the enriched phage population is determined by PCR amplification of the inserts, labeling, and hybridization to a yeast ORF microarray.</p
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