Paired-end mapping reveals extensive structural variation in the human genome

Structural variation of the genome involves kilobase- to megabase-sized deletions, duplications, insertions, inversions, and complex combinations of rearrangements. We introduce high-throughput and massive paired-end mapping (PEM), a large-scale genome-sequencing method to identify structural variants (SVs) approximately 3 kilobases (kb) or larger that combines the rescue and capture of paired ends of 3-kb fragments, massive 454 sequencing, and a computational approach to map DNA reads onto a reference genome. PEM was used to map SVs in an African and in a putatively European individual and identified shared and divergent SVs relative to the reference genome. Overall, we fine-mapped more than 1000 SVs and documented that the number of SVs among humans is much larger than initially hypothesized; many of the SVs potentially affect gene function. The breakpoint junction sequences of more than 200 SVs were determined with a novel pooling strategy and computational analysis. Our analysis provided insights into the mechanisms of SV formation in humans.     

Influence of electro-acupuncture on pain threshold in horses and its mode of action

Electro-acupuncture (EA) treatments varied in their effect on pain threshold in horses. EA stimulation using local acupuncture points or/and high frequency (80–120 Hz) can be more effective to relieve the experimental pain than the use of distal points and/or low frequency (20 Hz). The acupoints close to the painful areas may need to be stimulated with high frequency EA while the acupoints far from the painful areas may be stimulated with low frequency EA. The release of β-endorphin may be one of the pathways in which electro-acupuncture relieves the experimental pain.     

Identification of lidocaine and its metabolites in post-administration equine urine by ELISA and MS/MS

Lidocaine is a local anesthetic drug that is widely used in equine medicine. It has the advantage of giving good local anesthesia and a longer duration of action than procaine. Although approved for use in horses in training by the American Association of Equine Practitioners (AAEP), lidocaine is also an Association of Racing Commissioners International (ARCI) Class 2 drug and its detection in forensic samples can result in significant penalties. Lidocaine was observed as a monoprotonated ion at m/z 235 by ESI+ MS/MS (electrospray ionization-positive ion mode) analysis. The base peak ion at m/z 86, representing the postulated methylenediethylamino fragment [CH2N(CH2CH3)2]+, was characteristic of lidocaine and 3-hydroxylidocaine in both ESI+ and EI (electron impact-positive ion mode) mass spectrometry. In addition, we identified an ion at m/z 427 as the principal parent ion of the ion at m/z 86, consistent with the presence of a protonated analog of 3-hydroxylidocaine-glucuronide. We also sought to establish post-administration ELISA-based 'detection times' for lidocaine and lidocaine-related compounds in urine following single subcutaneous injections of various doses (10, 40, 400 mg). Our findings suggest relatively long ELISA based 'detection times' for lidocaine following higher doses of this drug.     

Developmentally regulated systemic endopolyploid in succulents with small genomes

Nuclei from Mesembryanthemum crystallinum (ice plant) exhibit multiple levels of ploidy in every tissue as revealed by flow microfluorometric analysis of isolated nuclei stained with mithramycin. Multiples of the haploid nuclear genome complement (1C) corresponding to 2C, 4C, 8C, 16C, 32C, and 64C were observed. The distribution of nuclei among the different ploidy levels is tissue-specific and in leaves is characteristic of the stage of development. This type of genome organization has been identified in eight other succulent CAM (crassulacean acid metabolism) plant species with small genomes. Multiploidy may be a common property of this type of plant.     

Ropivacaine in the horse: its pharmacological responses, urinary detection and mass spectral confirmation

This report evaluates the pharmacological responses, urinary detection and mass spectral confirmation of ropivacaine in horses. Ropivacaine, a potent local anesthetic (LA) recently introduced in human medicine, has an estimated highest no-effect dose (HNED) of about 0.4 mg/site as determined in our abaxial sesamoid block model. Apparent ropivacaine equivalents were detectable by ELISA screening using a mepivacaine ELISA test after administration of clinically effective doses. Mass spectral examination of postadministration urine samples showed no detectable parent ropivacaine, but a compound indistinguishable from authentic 3-hydroxyropivacaine was recovered from these samples. The study shows that ropivacaine is a potent LA in the horse, that clinically effective doses can be detected in postadministration samples by ELISA-based screening, and that its major post administration urinary metabolite is 3-hydroxyropivacaine.