Fri. May 24th, 2024

And, NZ; 3Department of Surgery, University of Auckland, Auckland, NZIntroduction: Tuberculous and non-tuberculous Mycobacteria release membrane CLEC-1 Proteins web vesicles (MMVs), reported to variety from 60 to 300 nm in diameter, predominantly contain lipoproteins and polar lipids. It’s hypothesised that MVs facilitate delivery of virulence elements and function as “immune decoys” modulating host immune responses contributing to severe disease. To better have an understanding of MMV biology we undertook the analysis of 3 species: Mycobacterium smegmatis (non-pathogenic, CD161/KLRB1 Proteins supplier fast-grower), M. abscessus (human pathogen, fast-grower) and M. marinum (fish and opportunistic human pathogen, slow-grower). The M. marinum-Saturday, May 20,zebrafish model has been proposed to become among the very best models to study human tuberculosis. Techniques and Results: Diverse MMV parameters including composition, size, concentration and release with respect to cell development and viability had been studied. Nanoparticle tracking analysis and electron microscopy techniques had been employed to figure out MMV concentration and size. We isolated MMVs with mean diameters amongst 8000 nm. SDSPAGE protein profiles were comparable for three isolations for every single species with interspecies differences. DNA and RNA concentrations amongst 25 and 35 /ml of original culture respectively were obtained. Conclusion: MMVs were developed throughout development, with most produced in the transition in between exponential and stationary phase. Stationary phase MMVs from M. abscessus were the largest ( 200 nm) and contained far 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 making use of live/dead staining, displaying that MMVs were developed by reside cells suggesting vesicle production may very well be an active biological process. Purification of MMVs by density gradient centrifugation showed distinct MMV rich fractions in all species investigated, with distinct DNA and RNA patterns across the density layers suggesting heterogeneity amongst 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 towards the patient before sample collection or mixture of purified EVs 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 usage of fluorescent metabolites for the detection of exosomes from cancer cells Alan M. Ezrin1, Michael W. Graner2 and Steven G. Griffiths1 NX Improvement 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 illness. Such diversity in shed vesicles endows the cell with all the ability to react to disparate physiological signals via the mobilisation of particular forms of vesicles. The two bestcharacterised classes of EVs at present are exosomes and microvesicles, distinguished largely on the basis of siz.