Living bacteria therapies have been proposed as an alternative approach to treating a broad array of cancers. In this study, we developed a genetically encoded microbial encapsulation system with tunable and dynamic expression of surface capsular polysaccharides that enhances systemic delivery. Based on a small RNA screen of capsular biosynthesis pathways, we constructed inducible synthetic gene circuits that regulate bacterial encapsulation in Escherichia coli Nissle 1917. These bacteria are capable of temporarily evading immune attack, whereas subsequent loss of encapsulation results in effective clearance in vivo. This dynamic delivery strategy enabled a ten-fold increase in maximum tolerated dose of bacteria and improved anti-tumor efficacy in murine models of cancer. Furthermore, in situ encapsulation increased the fraction of microbial translocation among mouse tumors, leading to efficacy in distal tumors. The programmable encapsulation system promises to enhance the therapeutic utility of living engineered bacteria for cancer.

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The sensitivity of insect nervous systems to anoxia can be modulated genetically and pharmacologically, but the cellular mechanisms responsible are poorly understood. We examined the effect of a heat shock pretreatment (HS) on the sensitivity of the locust (Locusta migratoria) nervous system to anoxia induced by water immersion. Prior HS made locusts more resistant to anoxia by increasing the time taken to enter a coma and by reducing the time taken to recover the ability to stand. Anoxic comas were accompanied by surges of extracellular potassium ions in the neuropile of the metathoracic ganglion, and HS reduced the time taken for clearance of excess extracellular potassium ions. This could not be attributed to a decrease in the activity of protein kinase G, which was increased by HS. In homogenates of the metathoracic ganglion, HS had only a mild effect on the activity of Na(+)-K(+)-ATPase. However, we demonstrated that HS caused a threefold increase in the immunofluorescent localization of the α-subunit of Na(+)-K(+)-ATPase in metathoracic neuronal plasma membranes relative to background labeling of the nucleus. We conclude that HS induced trafficking of Na(+)-K(+)-ATPase into neuronal plasma membranes and suggest that this was at least partially responsible for the increased resistance to anoxia and the increased rate of recovery of neural function after a disturbance of K(+) homeostasis.

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Prolonged life expectancy also results in an increased need for high-performance orthopedic implants. It has been shown that a compromised tissue-implant interface could lead to adverse immune-responses and even the dislodging of the implant. To overcome these obstacles, our research team has been seeking ways to decrease the risk of faulty tissue-implant interfaces by improving the biocompatibility and the osteo-inductivity of conventional orthopedic implants using ultrafine particle coatings. These particles were enriched with various bioactive additives prior to coating, and the coated biomaterial surfaces exhibited significantly increased biocompatibility and osteoinductivity. Physical assessments firstly confirmed the proper incorporation of the bioactive additives after examining their surface chemical composition. Then, in vitro assays demonstrated the biocompatibility and osteo-inductivity of the coated surfaces by studying the morphology of attached cells and their mineralization abilities. In addition, by quantifying the responses, activities and gene expressions, cellular evaluations confirmed the positive effects of these polymer based bioactive coatings. Consequently, the bioactive ultrafine polymer particles demonstrated their ability in improving the biocompatibility and osteo-inductivity of conventional orthopedic implants. As a result, our research team hope to apply this technology to the field of orthopedic implants by making them more effective medical devices through decreasing the risk of implant-induced immune responses and the loosening of the implant. OPEN ACCESSCoatings 2015, 5 738