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Obtaining accurate values of joint tissue loads in human subjects and animals in vivo requires exact 3D-reproduction of joint kinematics and comparisons of in vivo motions between subjects and animals, and also necessitates an accurate reference position. For the knee, passive flexion-extension of isolated joints by hand has been assumed to produce bony motions similar to those of normal gait. We hypothesized that passive flexion-extension kinematics would not accurately reproduce in vivo gait, and, further, that such kinematics would vary significantly between testers. In vivo gait motions of four ovine stifle joints were measured in six degrees of freedom, as were passive flexion-extension motions after sacrifice. Passive flexion-extension motions were performed by three testers on the same stifle joints used in vitro. Results showed statistically significant differences in all degrees of freedom, with the largest differences in the proximal-distal and internal-external directions. Differences induced by muscle loads and kinetic factors in vivo were most evident during stance and hoof-off phases of gait. The in vitro passive paths generated by hand created motions with large variability both between and within individual testers. The user dependence and \"area\" of motion of passive flexion-extension indicates that passive flexion-extension is contained in a volume of motion, rather than constrained to a unique path. The assumption that the passive path has relevance to precise bone positions during normal in vivo gait is not supported by these results. Thus, using passive flexion-extension as a reference between joints may introduce large motion variability in the observed outcome, and large potential errors in determining joint tissue loads.  相似文献   
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Predators influence the phenotype of prey through both natural selection and induction. We investigated the effects of grazers and nutrients on chain formation in a dinoflagellate, Cochlodinium polykrikoides, which forms dense blooms and has deleterious effects on marine ecosystems around the world. Field populations of C. polykrikoides formed longer chains than laboratory cultures without grazers. In the field, chain length of C. polykrikoides was positively correlated with the abundance of the copepod Acartia tonsa. Chain length of C. polykrikoides increased when exposed to live females of A. tonsa or its fresh (<24 h post-isolation) exudates for 48 h. These results suggest that dissolved chemical cues released by A. tonsa induce chain formation in C. polykrikoides. Ingestion rate of A. tonsa on four-cell chains of C. polykrikoides was lower than on single cells, suggesting that chain formation may be an effective anti-grazing defense. Finally, nutrient amendment experiments demonstrated that vitamins (B1, B7, and B12) increased the chain length of C. polykrikoides both singly and collectively, while trace metals and inorganic nutrients did not, showing that vitamins may also influence chain formation in this species.  相似文献   
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The synthesis and optimisation of HCV NS5B polymerase inhibitors with improved potency versus the existing compound 1 is described. Substitution in the benzothiadiazine portion of the molecule, furnishing improvement in potency in the high protein Replicon assay, is highlighted, culminating in the discovery of 12h, a highly potent oxyacetamide derivative.  相似文献   
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We have previously shown in cultured rat microvascular endothelial cells (RMEC) that lipopolysaccharide (LPS) stimulates a protein tyrosine kinase (PTK)-dependent reduction in cellular coupling. We hypothesized that connexin 43 (Cx43) becomes phosphorylated following exposure to LPS. Cx43 was immunoprecipitated from control and LPS-treated RMEC monolayers. Tyrosine phosphorylation of Cx43, detected by immunoblot, was found only in the LPS treatment. To verify these results, Cx43 was radiolabeled with [(32)P]-orthophosphate. Radiolabeled Cx43 exhibited a slight increase in phosphorylation in response to LPS; phosphoamino acid analysis displayed equivalent amounts of phosphoserine in control and LPS treatments, but detected phosphotyrosine only in the LPS treatment. The PTK inhibitors PP-2 (10 nM) and geldanamycin (200 nM) were found to block the response to LPS in terms of Cx43 tyrosine phosphorylation and cellular coupling. The phosphatase inhibitor BpV (1 microM) accentuated the effect of LPS, while the putative phosphatase activator C(6)-ceramide prevented it. When measuring cell communication, phosphatase inhibition also blocked the reversal of the LPS response following LPS washout. We conclude that Cx43 is tyrosine phosphorylated following exposure to LPS and suggest that the LPS-induced increase in intercellular resistance may be mediated by tyrosine phosphorylation of this connexin. Altering tyrosine kinase and phosphatase activities can modulate the LPS-induced tyrosine phosphorylation of Cx43 and reductions in cellular coupling.  相似文献   
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Fire shapes animal communities by altering resource availability and species interactions, including between predators and prey. In Australia, there is particular concern that two highly damaging invasive predators, the feral cat (Felis catus) and European red fox (Vulpes vulpes), increase their activity in recently burnt areas and exert greater predation pressure on the native prey due to their increased exposure. We tested how prescribed fire occurrence and extent, along with fire history, vegetation, topography, and distance to anthropogenic features (towns and farms), affected the activity (detection frequency) of cats, foxes, and the native mammal community in south-eastern Australia. We used camera traps to quantify mammal activity before and after a prescribed burn and statistically tested how the fire interacted with these habitat variables to affect mammal activity. We found little evidence that the prescribed fire influenced the activity of cats and foxes and no evidence of an effect on kangaroo or small mammal (<800 g) activity. Medium-sized mammals (800–2000 g) were negatively associated with prescribed fire extent, suggesting that prescribed fire has a negative impact on these species in the short term. The lack of a clear activity increase from cats and foxes is likely a positive outcome from a fire management perspective. However, we highlight that their response is likely dependent upon factors like fire size, severity, and prey availability. Future experiments should incorporate GPS-trackers to record fine-scale movements of cats and foxes in temperate ecosystems immediately before and after prescribed fire to best inform management within protected areas.  相似文献   
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Echinococcus multilocularis, the aetiological agent of human Alveolar Echinococcosis, is transmitted between small mammals and wild or domestic canids. Dogs infected with E. multilocularis as dead-end hosts. Whereas E. multilocularis infections in wild hosts and humans have been well-studied in recent decades, infections in domestic dogs are sparsely reported. This literature review and meta-analysis highlighted gaps in the available data and provided a re-assessment of the global distribution of domestic dog E. multilocularis infections. We found 46 published articles documenting the prevalence of E. multilocularis in domestic dogs from 21 countries across Europe, Asia and North America. Apparent prevalence estimates ranged from 0.00% (0.00–0.33%) in Germany to 55.50% (26.67–81.12%) in China. Most studies were conducted in areas of high human Alveolar Echinococcosis. By accounting for reassessed diagnostic sensitivity and specificity, we estimated true prevalence in a subset of studies, which varied between 0.00% (0.00–12.42%) and 41.09% (21.12–65.81%), as these true prevalence estimates were seldom reported in the articles themselves. Articles also showed a heavy emphasis on rural dogs, dismissing urban ones, which is concerning due to the role urbanisation plays in the transmission of zoonotic diseases, especially those utilising pets as definitive hosts. Lastly, population studies on canine Alveolar Echinococcosis were absent, highlighting the relative focus on human rather than animal health. We thus developed a framework for investigating domestic dog E. multilocularis infections and performing risk assessment of dog-associated transmission to fill the gaps found in the literature.  相似文献   
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Single molecule force spectroscopy is a powerful approach to probe the structure, conformational changes, and kinetic properties of biological and synthetic macromolecules. However, common approaches to apply forces to biomolecules require expensive and cumbersome equipment and relatively large probes such as beads or cantilevers, which limits their use for many environments and makes integrating with other methods challenging. Furthermore, existing methods have key limitations such as an inability to apply compressive forces on single molecules. We report a nanoscale DNA force spectrometer (nDFS), which is based on a DNA origami hinge with tunable mechanical and dynamic properties. The angular free energy landscape of the nDFS can be engineered across a wide range through substitution of less than 5% of the strand components. We further incorporate a removable strut that enables reversible toggling of the nDFS between open and closed states to allow for actuated application of tensile and compressive forces. We demonstrate the ability to apply compressive forces by inducing a large bend in a 249bp DNA molecule, and tensile forces by inducing DNA unwrapping of a nucleosome sample. These results establish a versatile tool for force spectroscopy and robust methods for designing nanoscale mechanical devices with tunable force application.  相似文献   
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