Minimizing and managing needle and instrument loss during pediatric robotic surgery.

INTRODUCTION: Lost objects and equipment malfunctions during robotic and laparoscopic cases can lead to an increase in operating time and potential risk to the patient. The literature on the management of foreign bodies during pediatric robotic-assisted surgery is limited. The purpose of the video is to review proper instrument handling to prevent loss of an object and to propose our technique for retrieving lost objects through two pediatric case examples. MATERIALS AND METHODS: The first case is a robotic-assisted laparoscopic left pyeloplasty in a 6-week-old male with congenital uteropelvic junction obstruction during which a needle was lost. In the video, we describe our techniques for safe needle passage, proper suturing technique, and recovering a lost needle. The second case is a robotic-assisted right upper pole heminephrectomy in a 14-month-old female with a duplicated renal collecting system and hydroureteronephrosis. We present the management of a rare case during which a harmonic scalpel jaw malfunctioned leading to a lost foreign body. We describe our technique for retrieving the lost fragment. RESULTS: All objects were safely removed, and patients were discharged post-op day one without complication. CONCLUSION: Our video presents two case examples of foreign bodies lost during pediatric robotic surgeries and approaches to manage each of these incidents.

Linkage between endosomal escape of LNP-mRNA and loading into EVs for transport to other cells.

RNA-based therapeutics hold great promise for treating diseases and lipid nanoparticles (LNPs) represent the most advanced platform for RNA delivery. However, the fate of the LNP-mRNA after endosome-engulfing and escape from the autophagy-lysosomal pathway remains unclear. To investigate this, mRNA (encoding human erythropoietin) was delivered to cells using LNPs, which shows, for the first time, a link between LNP-mRNA endocytosis and its packaging into extracellular vesicles (endo-EVs: secreted after the endocytosis of LNP-mRNA). Endosomal escape of LNP-mRNA is dependent on the molar ratio between ionizable lipids and mRNA nucleotides. Our results show that fractions of ionizable lipids and mRNA (1:1 molar ratio of hEPO mRNA nucleotides:ionizable lipids) of endocytosed LNPs were detected in endo-EVs. Importantly, these EVs can protect the exogenous mRNA during in vivo delivery to produce human protein in mice, detected in plasma and organs. Compared to LNPs, endo-EVs cause lower expression of inflammatory cytokines.