Symmetrical Dimethylarginine as a Diagnostic Parameter in Hermann's Tortoises (Testudo hermanni)

BackgroundDespite improvements in habitational conditions, kidney disease is relatively common in tortoises. ObjectivesPurpose of this study was the establishment of Symmetrical dimethylarginine (SDMA) reference values for clinically healthy Hermann's Tortoises. AnimalsClinically healthy Hermann's Tortoises (n = 131) were included in the period from October 2017 to September 2019. MethodsCreatinine and other biomarkers were tested at IDEXX Laboratories, Germany using residual blood samples from Hermann's tortoises. SDMA was measured with the IDEXX test and verified by liquid chromatography-mass spectrometry at IDEXX Laboratories, USA. ResultsSDMA values ranged from 1 to 21 mu g/dl (n = 131) for the IDEXX SDMA Test and SDMA values ranged from 1 to 17 mu g/dl (n = 82) for LC-MS. For the comparison of the two measuring systems, the following results were obtained R-2 = 0.75 (p < 0.001). Conclusion and Clinical ImportanceSDMA can be measured in Hermann's Tortoises and the reference values range in clinically healthy animals is comparable to that of dogs and cats

Expression capable library for studies of Neisseria gonorrhoeae, version 1.0

Background The sexually transmitted disease, gonorrhea, is a serious health problem in developed as well as in developing countries, for which treatment continues to be a challenge. The recent completion of the genome sequence of the causative agent, Neisseria gonorrhoeae, opens up an entirely new set of approaches for studying this organism and the diseases it causes. Here, we describe the initial phases of the construction of an expression-capable clone set representing the protein-coding ORFs of the gonococcal genome using a recombination-based cloning system. Results The clone set thus far includes 1672 of the 2250 predicted ORFs of the N. gonorrhoeae genome, of which 1393 (83%) are sequence-validated. Included in this set are 48 of the 61 ORFs of the gonococcal genetic island of strain MS11, not present in the sequenced genome of strain FA1090. L-arabinose-inducible glutathione-S-transferase (GST)-fusions were constructed from random clones and each was shown to express a fusion protein of the predicted size following induction, demonstrating the use of the recombination cloning system. PCR amplicons of each ORF used in the cloning reactions were spotted onto glass slides to produce DNA microarrays representing 2035 genes of the gonococcal genome. Pilot experiments indicate that these arrays are suitable for the analysis of global gene expression in gonococci. Conclusion This archived set of Gateway® entry clones will facilitate high-throughput genomic and proteomic studies of gonococcal genes using a variety of expression and analysis systems. In addition, the DNA arrays produced will allow us to generate gene expression profiles of gonococci grown in a wide variety of conditions. Together, the resources produced in this work will facilitate experiments to dissect the molecular mechanisms of gonococcal pathogenesis on a global scale, and ultimately lead to the determination of the functions of unknown genes in the genome

Radiation hybrid map spanning the huntington disease gene region of chromosome 4

Radiation hybrid (RH) mapping was used to construct a map of 11 markers in the distal 4 Mb of the short arm of chromosome 4, the region containing the Huntington disease gene. Two different methods for deriving the order of the markers were compared and both arrived at the same order as being the most likely. This order is also consistent with both the physical map constructed using pulsed-field gel electrophoresis (PFGE) and the meiotic linkage map. Comparing the RH map to the map determined by PFGE provided the means to equate RH map units (centirays) with actual physical distance in kilobases of DNA. In addition, a simple procedure for reducing the complexity of human DNA in radiation hybrids is described. One cell line isolated using this procedure contains, as its only human DNA, ~2 Mb surrounding the Huntington disease gene.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/29937/1/0000294.pd

Sequence-tagged sites (STSs) spanning 4p16.3 and the Huntington disease candidate region

The generation of sequence-tagged sites (STSs) has been proposed as a unifying approach to correlating the disparate results generated by genetic and various physical techniques being used to map the human genome. We have developed an STS map to complement the existing physical and genetic maps of 4p16.3, the region containing the Huntington disease gene. A total of 18 STSs span over 4 Mb of 4p16.3, with an average spacing of about 250 kb. Eleven of the STSs are located within the primary candidate HD region of 2.5 Mb between D4S126 and D4S168. The availability of STSs makes the corresponding loci accessible to the general community without the need for distribution of cloned DNA. These STSs should also provide the means to isolate yeast artificial chromosome clones spanning the HD candidate region.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30062/1/0000432.pd

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

The complete sequence of human chromosome 5

Chromosome 5 is one of the largest human chromosomes yet has one of the lowest gene densities. This is partially explained by numerous gene-poor regions that display a remarkable degree of noncoding and syntenic conservation with non-mammalian vertebrates, suggesting they are functionally constrained. In total, we compiled 177.7 million base pairs of highly accurate finished sequence containing 923 manually curated protein-encoding genes including the protocadherin and interleukin gene families and the first complete versions of each of the large chromosome 5 specific internal duplications. These duplications are very recent evolutionary events and play a likely mechanistic role, since deletions of these regions are the cause of debilitating disorders including spinal muscular atrophy (SMA)

Metabasic rocks from the Variscan Schwarzwald (SW Germany): metamorphic evolution and igneous protoliths

In the Variscan Schwarzwald metabasic rocks form small bodies included within anatectic plagioclase-biotite gneisses. Many metabasites first underwent an eclogite-facies metamorphism at about 2.0 GPa and 670–700 °C, resulting in the assemblage garnet + omphacite + rutile + quartz ± epidote ± amphibole ± kyanite. Since these eclogites are nearly free of an OH-bearing phase, they underwent almost complete dehydration during subduction, suggesting formation along an average to warm top-of-the-slab geotherm of 10–13 °C/km. The age of the Variscan high-P/high-T metamorphism is > 333 Ma. After partial exhumation from ~ 65 to ~ 15 km depth, the eclogites were overprinted under increasing activity of H2O by a number of retrograde reactions. The degree of this overprint under amphibolite-facies conditions (0.4–0.5 GPa/675–690 °C) was very different. Up to now, only retrograde eclogites have been found, but some samples still contain omphacite. Kyanite is at least partially transformed to aggregates of plagioclase + spinel ± corundum ± sapphirine. On the other hand, there are amphibolites that are extensively recrystallized and show the assemblage amphibole + plagioclase + ilmenite/titanite ± biotite ± quartz ± sulphides. The last relic phase that can be found in such otherwise completely recrystallized amphibolites is rutile. After the amphibolite-facies metamorphism at ~ 333 Ma, the metabasites underwent a number of low-temperature transformations, such as sericitization of plagioclase, chloritization of amphibole, and formation of prehnite. The intimate association of metabasite bodies with gneisses of dominantly meta-greywacke compositions suggests derivation from an active plate margin. This view is corroborated by bulk-rock geochemical data. Excluding elements that were mobile during metamorphism (Cs, Rb, Ba, K, Pb, Sr, U), the concentrations of the remaining elements in most of the metabasites are compatible with a derivation from island-arc tholeiites, back-arc basin basalts or calc-alkaline basalts. Only some samples have MORB precursor rocks.Ruprecht-Karls-Universität Heidelberg (1026