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Kandasamy, Saveetha,Khan, Wajahatullah,Kulshreshtha, Garima,Evans, Franklin,Critchley, Alan T.,Fitton, J.H.,Stringer, Damien N.,Gardiner, Vicki-Anne,Prithiviraj, Balakrishnan The Korean Society of Phycology 2015 ALGAE Vol.30 No.2
Brown algal extracts have long been used as feed supplements to promote health of farm animals. Here, we show new molecular insights in to the mechanism of action of a fucose containing polymer (FCP) rich fraction from the brown seaweed Ascophyllum nodosum using the Caenorhabditis elegans-Pseudomonas aeruginosa PA14 infection model. FCP enhanced survival of C. elegans against pathogen stress, correlated with up-regulation of key immune response genes such as: lipases, lysozyme (lys-1), saponin-like protein (spp-1), thaumatin-like protein (tlp-1), matridin SK domain protein (msk-1), antibacterial protein (abf-1), and lectin family protein (lfp). Further, FCP caused down regulation of P. aeruginosa quorum sensing genes: (lasI, lasR, rhlI, and rhlR), secreted virulence factors (lipase, proteases, and elastases) and toxic metabolites (pyocyanin, hydrogen cyanide, and siderophore). Biofilm formation and motility of pathogenic bacteria were also greatly attenuated when the culture media were treated with FCP. Interestingly, FCP failed to mitigate the pathogen stress in skn-1, daf-2, and pmk-1 mutants of C. elegans. This indicated that, FCP treatment acted on the regulation of fundamental innate immune pathways, which are conserved across the majority of organisms including humans. This study suggests the possible use of FCP, a seaweed component, as a functional food source for healthy living.
Saveetha Kandasamy,Wajahatullah Khan,Garima Kulshreshtha,Franklin Evans,Alan T. Critchley,J. H. Fitton,Damien N. Stringer,Vicki-Anne Gardiner,Balakrishnan Prithiviraj 한국조류학회I 2015 ALGAE Vol.30 No.2
Brown algal extracts have long been used as feed supplements to promote health of farm animals. Here, we show new molecular insights in to the mechanism of action of a fucose containing polymer (FCP) rich fraction from the brown seaweed Ascophyllum nodosum using the Caenorhabditis elegans-Pseudomonas aeruginosa PA14 infection model. FCP enhanced survival of C. elegans against pathogen stress, correlated with up-regulation of key immune response genes such as: lipases, lysozyme (lys-1), saponin-like protein (spp-1), thaumatin-like protein (tlp-1), matridin SK domain protein (msk-1), antibacterial protein (abf-1), and lectin family protein (lfp). Further, FCP caused down regulation of P. aeruginosa quorum sensing genes: (lasI, lasR, rhlI, and rhlR), secreted virulence factors (lipase, proteases, and elastases) and toxic metabolites (pyocyanin, hydrogen cyanide, and siderophore). Biofilm formation and motility of pathogenic bacteria were also greatly attenuated when the culture media were treated with FCP. Interestingly, FCP failed to mitigate the pathogen stress in skn-1, daf-2, and pmk-1 mutants of C. elegans. This indicated that, FCP treatment acted on the regulation of fundamental innate immune pathways, which are conserved across the majority of organisms including humans. This study suggests the possible use of FCP, a seaweed component, as a functional food source for healthy living.
Neurosphere and adherent culture conditions are equivalent for malignant glioma stem cell lines
Maryam Rahman,Karina Reyner,Loic Deleyrolle,Sebastien Millette,Hassan Azari,Bryan W. Day,Brett W. Stringer,Andrew W. Boyd,Terrance G. Johns,Vincent Blot,Rohit Duggal,Brent A. Reynolds 대한해부학회 2015 Anatomy & Cell Biology Vol.48 No.1
Certain limitations of the neurosphere assay (NSA) have resulted in a search for alternative culture techniques forbrain tumor-initiating cells (TICs). Recently, reports have described growing glioblastoma (GBM) TICs as a monolayer usinglaminin. We performed a side-by-side analysis of the NSA and laminin (adherent) culture conditions to compare the growthand expansion of GBM TICs. GBM cells were grown using the NSA and adherent culture conditions. Comparisons weremade using growth in culture, apoptosis assays, protein expression, limiting dilution clonal frequency assay, genetic affymetrixanalysis, and tumorigenicity in vivo. In vitro expansion curves for the NSA and adherent culture conditions were virtuallyidentical (P=0.24) and the clonogenic frequencies (5.2% for NSA vs. 5.0% for laminin, P=0.9) were similar as well. Likewise,markers of differentiation (glial fibrillary acidic protein and beta tubulin III) and proliferation (Ki67 and MCM2) revealedno statistical difference between the sphere and attachment methods. Several different methods were used to determine thenumbers of dead or dying cells (trypan blue, DiIC, caspase-3, and annexin V) with none of the assays noting a meaningfulvariance between the two methods. In addition, genetic expression analysis with microarrays revealed no significant differencesbetween the two groups. Finally, glioma cells derived from both methods of expansion formed large invasive tumors exhibitingGBM features when implanted in immune-compromised animals. A detailed functional, protein and genetic characterization ofhuman GBM cells cultured in serum-free defined conditions demonstrated no statistically meaningful differences when grownusing sphere (NSA) or adherent conditions. Hence, both methods are functionally equivalent and remain suitable options forexpanding primary high-grade gliomas in tissue culture.