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Recent Submissions

  • Item type: Item ,
    Corneal epithelial cells exposed to shear stress show altered cytoskeleton and migratory behaviour
    (Public Library of Science, 2017-06-29) Molladavoodi, Sara; Robichaud, Matthew; Wulff, David; Gorbet, Maud
    Cells that form the corneal epithelium, the outermost layer of the cornea, are exposed to shear stress through blinking during waking hours. In this in vitro study, the effect of fluid shear stress on human corneal epithelial cells (HCECs) was investigated. Following exposure to shear stresses of 4 and 8 dyn/cm2, HCECs showed cytoskeletal rearrangement with more prominent, organized and elongated filamentous actin. Cytoskeletal changes were time-dependent, and were most significant after 24 hours of shear stress. Higher rates of migration and proliferation, as evaluated by a scratch assay, were also observed following 24 hours of low shear stress exposure (4 dyn/cm2). This result contrasted the poor migration observed in samples scratched before shear exposure, indicating that shear-induced cytoskeletal changes played a key role in improved wound healing and must therefore precede any damage to the cell layer. HCEC cytoskeletal changes were accompanied by an upregulation in integrin β1 and downregulation of ICAM-1. These results demonstrate that HCECs respond favourably to flow-induced shear stress, impacting their proliferation and migration properties as well as phenotype.
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    Climate change and marine fisheries: Least developed countries top global index of vulnerability
    (Public Library of Science, 2017-06-20) Blasiak, Robert; Spijkers, Jessica; Tokunaga, Kanae; Pittman, Jeremy; Yagi, Nobuyuki; Osterblom, Henrik
    Future impacts of climate change on marine fisheries have the potential to negatively influence a wide range of socio-economic factors, including food security, livelihoods and public health, and even to reshape development trajectories and spark transboundary conflict. Yet there is considerable variability in the vulnerability of countries around the world to these effects. We calculate a vulnerability index of 147 countries by drawing on the most recent data related to the impacts of climate change on marine fisheries. Building on the Intergovernmental Panel on Climate Change framework for vulnerability, we first construct aggregate indices for exposure, sensitivity and adaptive capacity using 12 primary variables. Seven out of the ten most vulnerable countries on the resulting index are Small Island Developing States, and the top quartile of the index includes countries located in Africa (17), Asia (7), North America and the Caribbean (4) and Oceania (8). More than 87% of least developed countries are found within the top half of the vulnerability index, while the bottom half includes all but one of the Organization for Economic Co-operation and Development member states. This is primarily due to the tremendous variation in countries’ adaptive capacity, as no such trends are evident from the exposure or sensitivity indices. A negative correlation exists between vulnerability and per capita carbon emissions, and the clustering of states at different levels of development across the vulnerability index suggests growing barriers to meeting global commitments to reducing inequality, promoting human well-being and ensuring sustainable cities and communities. The index provides a useful tool for prioritizing the allocation of climate finance, as well as activities aimed at capacity building and the transfer of marine technology.
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    Continental divide: Predicting climate-mediated fragmentation and biodiversity loss in the boreal forest
    (Public Library of Science, 2017-05-15) Murray, Dennis L.; Peers, Michael J. L.; Majchrzak, Yasmine N.; Wehtje, Morgan; Ferreira, Catarina; Rickles, Rob S. A.; Row, Jeffrey R.; Thornton, Daniel H.
    Climate change threatens natural landscapes through shifting distribution and abundance of species and attendant change in the structure and function of ecosystems. However, it remains unclear how climate-mediated variation in species’ environmental niche space may lead to large-scale fragmentation of species distributions, altered meta-population dynamics and gene flow, and disrupted ecosystem integrity. Such change may be especially relevant when species distributions are restricted either spatially or to a narrow environmental niche, or when environments are rapidly changing. Here, we use range-wide environmental niche models to posit that climate-mediated range fragmentation aggravates the direct effects of climate change on species in the boreal forest of North America. We show that climate change will directly alter environmental niche suitability for boreal-obligate species of trees, birds and mammals (n = 12), with most species ranges becoming smaller and shifting northward through time. Importantly, species distributions will become increasingly fragmented, as characterized by smaller mean size and greater isolation of environmentally-suitable landscape patches. This loss is especially pronounced along the Ontario-Québec border, where the boreal forest is narrowest and roughly 78% of suitable niche space could disappear by 2080. Despite the diversity of taxa surveyed, patterns of range fragmentation are remarkably consistent, with our models predicting that spruce grouse (Dendragapus canadensis), boreal chickadee (Poecile hudsonicus), moose (Alces americanus) and caribou (Rangifer tarandus) could have entirely disjunct east-west population segments in North America. These findings reveal potentially dire consequences of climate change on population continuity and species diversity in the boreal forest, highlighting the need to better understand: 1) extent and primary drivers of anticipated climate-mediated range loss and fragmentation; 2) diversity of species to be affected by such change; 3) potential for rapid adaptation in the most strongly-affected areas; and 4) potential for invasion by replacement species.
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    Sarcolipin deletion exacerbates soleus muscle atrophy and weakness in phospholamban overexpressing mice
    (Public Library of Science, 2017-03-09) Fajardo, Val A.; Gamu, Daniel; Mitchell, Andrew; Bloemberg, Darin; Bombardier, Eric; Chambers, Paige J.; Bellissimo, Catherine; Quadrilatero, Joe; Tuplin, A. Russell
    Sarcolipin (SLN) and phospholamban (PLN) are two small proteins that regulate the sarco(endo)plasmic reticulum Ca2+-ATPase pumps. In a recent study, we discovered that Pln overexpression (PlnOE) in slow-twitch type I skeletal muscle fibers drastically impaired SERCA function and caused a centronuclear myopathy-like phenotype, severe muscle atrophy and weakness, and an 8 to 9-fold upregulation of SLN protein in the soleus muscles. Here, we sought to determine the physiological role of SLN upregulation, and based on its role as a SERCA inhibitor, we hypothesized that it would represent a maladaptive response that contributes to the SERCA dysfunction and the overall myopathy observed in the PlnOE mice. To this end, we crossed Sln-null (SlnKO) mice with PlnOE mice to generate a PlnOE/SlnKO mouse colony and assessed SERCA function, CNM pathology, in vitro contractility, muscle mass, calcineurin signaling, daily activity and food intake, and proteolytic enzyme activity. Our results indicate that genetic deletion of Sln did not improve SERCA function nor rescue the CNM phenotype, but did result in exacerbated muscle atrophy and weakness, due to a failure to induce type II fiber compensatory hypertrophy and a reduction in total myofiber count. Mechanistically, our findings suggest that impaired calcineurin activation and resultant decreased expression of stabilin-2, and/or impaired autophagic signaling could be involved. Future studies should examine these possibilities. In conclusion, our study demonstrates the importance of SLN upregulation in combating muscle myopathy in the PlnOE mice, and since SLN is upregulated across several myopathies, our findings may reveal SLN as a novel and universal therapeutic target.
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    Functional metagenomics reveals novel β-galactosidases not predictable from gene sequences
    (Public Library of Science, 2017-03-08) Cheng, Jiujun; Romantsov, Tatyana; Engel, Katja; Doxey, Andrew C.; Rose, David R.; Neufeld, Josh D.; Charles, Trevor C.
    The techniques of metagenomics have allowed researchers to access the genomic potential of uncultivated microbes, but there remain significant barriers to determination of gene function based on DNA sequence alone. Functional metagenomics, in which DNA is cloned and expressed in surrogate hosts, can overcome these barriers, and make important contributions to the discovery of novel enzymes. In this study, a soil metagenomic library carried in an IncP cosmid was used for functional complementation for β-galactosidase activity in both Sinorhizobium meliloti (α-Proteobacteria) and Escherichia coli (γ-Proteobacteria) backgrounds. One β-galactosidase, encoded by six overlapping clones that were selected in both hosts, was identified as a member of glycoside hydrolase family 2. We could not identify ORFs obviously encoding possible β-galactosidases in 19 other sequenced clones that were only able to complement S. meliloti. Based on low sequence identity to other known glycoside hydrolases, yet not β-galactosidases, three of these ORFs were examined further. Biochemical analysis confirmed that all three encoded β-galactosidase activity. Lac36W_ORF11 and Lac161_ORF7 had conserved domains, but lacked similarities to known glycoside hydrolases. Lac161_ORF10 had neither conserved domains nor similarity to known glycoside hydrolases. Bioinformatic and structural modeling implied that Lac161_ORF10 protein represented a novel enzyme family with a five-bladed propeller glycoside hydrolase domain. By discovering founding members of three novel β-galactosidase families, we have reinforced the value of functional metagenomics for isolating novel genes that could not have been predicted from DNA sequence analysis alone.