And, NZ; 3Department of Surgery, University of Auckland, Auckland, NZIntroduction: Tuberculous and non-tuberculous Mycobacteria release membrane vesicles (MMVs), reported to range from 60 to 300 nm in diameter, predominantly contain lipoproteins and polar lipids. It’s hypothesised that MVs facilitate delivery of virulence aspects and function as “immune decoys” modulating host immune responses contributing to extreme disease. To much better comprehend MMV biology we undertook the analysis of 3 species: Mycobacterium smegmatis (non-pathogenic, fast-grower), M. abscessus (human pathogen, fast-grower) and M. marinum (fish and opportunistic human pathogen, slow-grower). The M. marinum-Saturday, May well 20,zebrafish model has been proposed to be one of many most effective models to study human tuberculosis. Methods and Outcomes: Diverse MMV parameters such as composition, size, concentration and release with respect to cell growth and viability have been studied. Nanoparticle tracking evaluation and electron microscopy procedures were applied to identify MMV concentration and size. We isolated MMVs with imply diameters in between 8000 nm. SDSPAGE protein profiles have been related for three isolations for every single species with interspecies variations. DNA and RNA concentrations involving 25 and 35 /ml of original culture respectively were obtained. Conclusion: MMVs have been created throughout growth, with most developed at the transition between exponential and stationary phase. Stationary phase MMVs from M. abscessus have been the largest ( 200 nm) and contained much more DNA than RNA ( 20 suggesting the existence of a selective packaging mechanism. MMVs from M. smegmatis and M. marinum contained equal levels of DNA and RNA. MMV production was correlated with cell viability using live/dead staining, displaying that MMVs have been developed by live cells suggesting vesicle production could possibly be an active biological approach. Purification of MMVs by density gradient centrifugation showed distinct MMV rich fractions in all species investigated, with diverse DNA and RNA patterns across the density layers suggesting heterogeneity among species. In vitro experiments difficult THP-1 cells with M. marinum vesicles showed that MMVs had a dose dependent effect on THP-1 cell viability. Further investigation is required to recognize the active MMV elements, the mechanism of killing and to characterise the effects of sub-lethal MMV challenges.Gliolan to the patient prior to sample collection or mixture of purified EVs soon after collection.PS04.Identification of a novel population of lipid-rich extracellular vesicles Alanna Sedgwick1, M. Olivia Balmert1 and Crislyn D’Souza-SchoreyUniversity of Notre Dame, IL, USA; 2Department of Biological Sciences, University of Notre Dame, IL, USAPS04.The use of fluorescent metabolites for the detection of exosomes from cancer cells Alan M. Ezrin1, Michael W. Graner2 and Steven G. Griffiths1 NX Development Ubiquitin Conjugating Enzyme E2 L3 Proteins site Corporation; 2University of Colorado Denver, Anschutz Healthcare Campus, Dept of Neurosurgery, CO, USA; 3X0S0MEExtracellular vesicles (EVs) comprise a heterogeneous group of cargoloaded vesicles, that are released from cells to mediate extracellular communication in typical physiology and disease. Such diversity in shed vesicles endows the cell with the ability to react to Frizzled-4 Proteins supplier disparate physiological signals through the mobilisation of distinct types of vesicles. The two bestcharacterised classes of EVs at present are exosomes and microvesicles, distinguished largely around the basis of siz.