3'-UTR Sequence of Exosomal NANOGP8 DNA as an Extracellular Vesicle-Localization Signal.

Extracellular vesicles (EVs) are garnering attention as a safe and efficient biomolecule delivery system. EVs intrinsically play a crucial role in intercellular communication and pathophysiology by transporting functionally active DNA molecules. The internalized DNA pleiotropically affects the recipient cells. Considering these salient features, an intentional incorporation of specific DNA gene cassettes into EVs and their subsequent delivery to the target cells has potential applications in genetic engineering. Moreover, efficient ways to insert the DNA into EVs during their biogenesis is valuable. Our current research is a step in the development of this technology. As such, cancer cells are known to secrete exosomes containing increased amounts of double-stranded DNA than normal cells. The clonal analysis in our previously published data revealed that exosomes released from various cancer cells contained a significantly larger population of NANOGP8 DNA with a 22-base pair insertion in the 3'-untranslated region (UTR) compared to those secreted by normal cells. This finding led us to hypothesize that the 22-base pair insertion may act as a signal to facilitate the incorporation of NANOGP8 DNA into the exosomes. To test this hypothesis, we compared the EV localization of an Enhanced Green Fluorescent Protein (EGFP) gene fused with the NANOGP8 3'-UTR, with and without the 22-base pair insertion. The quantitative PCR analysis showed a significantly higher EGFP DNA accumulation in exosomes released from cells transfected with the gene cassette containing the 3'-UTR with the 22-base pair insertion. The discovery of a DNA localization signal in exosomal DNA's 3'-UTR could pave the way for the development of an EV-based DNA delivery system. This technology will open new possibilities in genetic engineering and innovative therapies using nucleic acid medicine.

Cost Savings in Urology Operating Rooms by Editing Surgeon Preference Cards.

BACKGROUND AND OBJECTIVES: Operating room costs contribute significantly to the overall expenditure for inpatient care. We evaluated a simple way to reduce urology operating room costs by limiting the loss from unused disposable items. METHODS: Baseline data were collected on opened and unused disposable items. Surgeons were asked to edit their preference cards and mark optional surgical items that would only be opened if requested. RESULTS: The cost of unused disposable items during the first 4 weeks in 3 operating rooms averaged $410/week. Costs after implementing the intervention declined to an average of $30/week. This yielded $380/week in savings, equating to a 92% reduction in waste, and a potential savings of $19 760 annually in the 2 urology operating rooms alone. CONCLUSION: Since the urology department represents only 10% of the main operating rooms at our institution, if other operating rooms implemented similar cost saving methods the hospital could potentially accrue significant savings.