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861.
Kobayashi T Antar AA Boehme KW Danthi P Eby EA Guglielmi KM Holm GH Johnson EM Maginnis MS Naik S Skelton WB Wetzel JD Wilson GJ Chappell JD Dermody TS 《Cell host & microbe》2007,1(2):147-157
Mammalian orthoreoviruses (reoviruses) are highly tractable experimental models for studies of double-stranded (ds) RNA virus replication and pathogenesis. Reoviruses infect respiratory and intestinal epithelium and disseminate systemically in newborn animals. Until now, a strategy to rescue infectious virus from cloned cDNA has not been available for any member of the Reoviridae family of dsRNA viruses. We report the generation of viable reovirus following plasmid transfection of murine L929 (L) cells using a strategy free of helper virus and independent of selection. We used the reovirus reverse genetics system to introduce mutations into viral capsid proteins sigma1 and sigma3 and to rescue a virus that expresses a green fluorescent protein (GFP) transgene, thus demonstrating the tractability of this technology. The plasmid-based reverse genetics approach described here can be exploited for studies of reovirus replication and pathogenesis and used to develop reovirus as a vaccine vector. 相似文献
862.
863.
Anderson C Bartlett SJ Gansner JM Wilson D He L Gitlin JD Kelsh RN Dowden J 《Molecular bioSystems》2007,3(1):51-59
As a result of a chemical genetic screen for modulators of metalloprotease activity, we report that 2-mercaptopyridine-N-oxide induces a conspicuous undulating notochord defect in zebrafish embryos, a phenocopy of the leviathan mutant. The location of the chemically-induced wavy notochord correlated with the timing of application, thus defining a narrow chemical sensitivity window during segmentation stages. Microscopic observations revealed that notochord undulations appeared during the phase of notochord cell vacuolation and notochord elongation. Notochord cells become swollen as well as disorganized, while electron microscopy revealed disrupted organization of collagen fibrils in the surrounding sheath. We demonstrate by assay in zebrafish extracts that 2-mercaptopyridine-N-oxide inhibits lysyl oxidase. Thus, we provide insight into notochord morphogenesis and reveal novel compounds for lysyl oxidase inhibition. Taken together, these data underline the utility of small molecules for elucidating the dynamic mechanisms of early morphogenesis and provide a potential explanation for the recently established role of copper in zebrafish notochord formation. 相似文献
864.
In recent years, the glycoconjugates of many parasitic nematodes have attracted interest due to their immunogenic and immunomodulatory nature. Previous studies with the porcine roundworm parasite Ascaris suum have focused on its glycosphingolipids, which were found, in part, to be modified by phosphorylcholine. Using mass spectrometry and western blotting, we have now analyzed the peptide N-glycosidase A-released N-glycans of adults of this species. The presence of hybrid bi- and triantennary N-glycans, some modified by core alpha1,6-fucose and peripheral phosphorylcholine, was demonstrated by LC/electrospray ionization (ESI)-Q-TOF-MS/MS, as was the presence of paucimannosidic N-glycans, some of which carry core alpha1,3-fucose, and oligomannosidic oligosaccharides. Western blotting verified the presence of protein-bound phosphorylcholine and core alpha1,3-fucose, whereas glycosyltransferase assays showed the presence of core alpha1,6-fucosyltransferase and Lewis-type alpha1,3-fucosyltransferase activities. Although, the unusual tri- and tetrafucosylated glycans found in the model nematode Caenorhabditis elegans were not found, the vast majority of the N-glycans found in A. suum represent a subset of those found in C. elegans; thus, our data demonstrate that the latter is an interesting glycobiological model for parasitic nematodes. 相似文献
865.
Ontogenetic colour change is typically associated with changes in size, vulnerability or habitat, but assessment of its functional significance requires quantification of the colour signals from the receivers' perspective. The tropical python, Morelia viridis, is an ideal species to establish the functional significance of ontogenetic colour change. Neonates hatch either yellow or red and both the morphs change to green with age. Here, we show that colour change from red or yellow to green provides camouflage from visually oriented avian predators in the different habitats used by juveniles and adults. This reflects changes in foraging behaviour and vulnerability as individuals mature and provides a rare demonstration of the adaptive value of ontogenetic colour change. 相似文献
866.
Patel K Lewiston D Gu Y Hicks KO Wilson WR 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2007,856(1-2):302-311
PR-104 is a dinitrobenzamide mustard pre-prodrug that is activated by reduction to a cytotoxic hydroxylamine metabolite in hypoxic tumour cells; it has recently commenced Phase I clinical trial. Here, we report two validated methods for the determination of PR-104 and its alcohol hydrolysis product, PR-104A in plasma and tissues across species. A high pH LC/MS/MS method was optimised for rapid and sensitive analysis of both analytes in rat, dog and human plasma. This assay was linear over the concentration range 0.005-2.5 microg/ml for PR-104 and 0.05-25 microg/ml for PR-104A (0.005-2.5 microg/ml for rat). A second method, using a low pH LC separation, was designed to provide higher chromatographic resolution, facilitating identification of metabolites. Both methods were successfully applied to the plasma pharmacokinetics of PR-104 and PR-104A in rats. In addition, cytotoxic reduced metabolites of PR-104A were identified in human tumour xenografts by the higher chromatographic resolution method. 相似文献
867.
M.J. Siegert A. Behar M. Bentley D. Blake S. Bowden P. Christoffersen C. Cockell H. Corr D. C. Cullen H. Edwards A. Ellery C. Ellis-Evans G. Griffiths R. Hindmarsh D. A. Hodgson E. King H. Lamb L. Lane K. Makinson M. Mowlem J. Parnell D. A. Pearce J. Priscu A. Rivera M. A. Sephton M. R. Sims A . M. Smith M. Tranter J. L. Wadham G. Wilson J. Woodward 《Reviews in Environmental Science and Biotechnology》2007,6(1-3):161-179
Antarctic subglacial lakes have, over the past few years, been hypothesised to house unique forms of life and hold detailed
sedimentary records of past climate change. Testing this hypothesis requires in situ examinations. The direct measurement
of subglacial lakes has been considered ever since the largest and best-known lake, named Lake Vostok, was identified as having
a deep water-column. The Subglacial Antarctic Lake Environments (SALE) programme, set up by the Scientific Committee on Antarctic
Research (SCAR) to oversee subglacial lakes research, state that prior exploration of smaller lakes would be a “prudent way
forward”. Over 145 subglacial lakes are known to exist in Antarctica, but one lake in West Antarctica, officially named Ellsworth
Subglacial Lake (referred to hereafter as Lake Ellsworth), stands out as a candidate for early exploration. A consortium of
over 20 scientists from seven countries and 14 institutions has been assembled to plan the exploration of Lake Ellsworth.
An eight-year programme is envisaged: 3 years for a geophysical survey, 2 years for equipment development and testing, 1 year
for field planning and operation, and 2 years for sample analysis and data interpretation. The science experiment is simple
in concept but complex in execution. Lake Ellsworth will be accessed using hot water drilling. Once lake access is achieved,
a probe will be lowered down the borehole and into the lake. The probe will contain a series of instruments to measure biological,
chemical and physical characteristics of the lake water and sediments, and will utilise a tether to the ice surface through
which power, communication and data will be transmitted. The probe will pass through the water column to the lake floor. The
probe will then be pulled up and out of the lake, measuring its environment continually as this is done. Once at the ice surface,
any water samples collected will be taken from the probe for laboratory analysis (to take place over subsequent years). The
duration of the science mission, from deployment of the probe to its retrieval, is likely to take between 24 and 36 h. Measurements
to be taken by the probe will provide data about the following: depth, pressure, conductivity and temperature; pH levels;
biomolecules (using life marker chips); anions (using a chemical analyzer); visualisation of the environment (using cameras
and light sources); dissolved gases (using chromatography); and morphology of the lake floor and sediment structures (using
sonar). After the probe has been retrieved, a sediment corer may be dropped into the lake to recover material from the lake
floor. Finally, if time permits, a thermistor string may be left in the lake water to take time-dependent measurements of
the lake’s water column over subsequent years. Given that the comprehensive geophysical survey of the lake will take place
in two seasons during 2007–2009, a two-year instrument and logistic development phase from 2008 (after the lake’s bathymetry
has been assessed) makes it possible that the exploration of Lake Ellsworth could take place at the beginning of the next
decade. 相似文献
868.
Lourenço WR 《Comptes rendus biologies》2007,330(10):770-777
A new genus and species of scorpion belonging to the recently named family Pseudochactidae is described based on two specimens collected in the Tham Xe Bangfai cave, Province of Khammouan, Laos. This new scorpion represents the second known record of a pseudochactid, and the first from Laos. The possible biogeographic consequences of this scorpion's distribution are discussed, associated with the possible Pangaean origin of pseudochactid scorpions. 相似文献
869.
Mark C. Ball Richard Pither Micheline Manseau Jeff Clark Stephen D. Petersen Steve Kingston Natasha Morrill Paul Wilson 《Conservation Genetics》2007,8(3):577-586
Faecal material has increasingly become an important non-invasive source of DNA for wildlife population genetics. However,
DNA from faecal sources can have issues associated with quantity (low-template and/or low target-to-total DNA ratio) and quality (degradation and/or low DNA-to-inhibitor ratio). A number of studies utilizing faecal material assume
and compensate for the above properties with minimal characterization of quantity or quality of target DNA, which can unnecessarily increase the risk of downstream technical problems. Here, we present a protocol which quantifies
faecal DNA using a two step approach: (1) estimating total DNA concentration using a PicogreenTM fluorescence assay and (2) estimating target nuclear DNA concentration by comparing amplification products of field samples at suspected concentrations to those of control
DNA at known concentrations. We applied this protocol to faecal material collected in the field from two species: woodland
caribou (Rangifer tarandus) and swift fox (Vulpes velox). Total DNA estimates ranged from 6.5 ng/μl to 28.6 ng/μl (X = 16.2 ng/μl) for the caribou extracts and 1.0–26.1 ng/μl (X = 7.5 ng/μl) for the swift fox extracts. Our results showed high concordance between total and target DNA estimates from woodland caribou faecal extracts, with only 10% of the samples showing relatively lower target-to-total DNA ratios. In contrast, DNA extracts from swift fox scat exhibited low target DNA yields, with only 38% (19 of 50) of the samples showing comparative target DNA amplification of at least 0.1 ng. With this information, we were able to estimate the amount of target DNA entered into PCR amplifications, and identify samples having target DNA below a lower threshold of 0.2 ng and requiring modification to genotyping protocols such as multiple tube amplification.
Our results here also show that this approach can easily be adapted to other species where faeces are the primary source of
DNA template. 相似文献
870.
Stephen D. Petersen Tom Mason Sarah Akber Rick West Brad White Paul Wilson 《Conservation Genetics》2007,8(2):497-502
This paper outlines a novel, non-invasive procedure to obtain DNA from Mexican tarantulas (Brachypelma spp.) using exuvia. These species are important in the pet trade and species identification is important for international
wildlife law enforcement. Mitochondrial DNA sequence from the cytochrome c oxidase subunit I gene was used to investigate the relationship between various Brachypelma spp. This phylogeny was used as a framework to assign unknown specimens and spiderlings to species. The benefits to conservation,
research, and international wildlife law enforcement that are gained by the ability to accurately identify species without
the death of the specimen are explored. Our data also suggest that there is no support for the genus Brachypelmides as some authors have proposed and upholds the synonymy of Locht et al. (1999) J Arachnol 27:196–200. 相似文献