Robotic surgery training: construct validity of Global Evaluative Assessment of Robotic Skills (GEARS)

The objective of this study is to determine the ability of the GEARS scale (Global Evaluative Assessment of Robotic Skills) to differentiate individuals with different levels of experience in robotic surgery, as a fundamental validation. This is a cross-sectional study that included three groups of individuals with different levels of experience in robotic surgery (expert, intermediate, novice) their performance were assessed by GEARS applied by two reviewers. The difference between groups was determined by Mann–Whitney test and the consistency between the reviewers was studied by Kendall W coefficient. The agreement between the reviewers of the scale GEARS was 0.96. The score was 29.8 ± 0.4 to experts, 24 ± 2.8 to intermediates and 16 ± 3 to novices, with a statistically significant difference between all of them (p < 0.05). All parameters from the scale allow discriminating between different levels of experience, with exception of the depth perception item. We conclude that the scale GEARS was able to differentiate between individuals with different levels of experience in robotic surgery and, therefore, is a validated and useful tool to evaluate surgeons in training.     

A lysolecithin/fatty acid mixture promotes and then blocks neurotransmitter release at the Drosophila melanogaster larval neuromuscular junction

The study of the effect of snake presynaptic neurotoxins with phospholipase A2 activity on nerve terminals has recently unveiled the inhibitory action of a lysophosphatidylcholine (LysoPC)/fatty acid mixture. We report here that these neurotoxins have no activity on Drosophila melanogaster nerve terminals. However, a 1:1 mixture of LysoPC and oleic acid induces an early increase, followed by an inhibition of both evoked and spontaneous neurotransmitter release. This effect is also induced by LysoPC alone. The present findings provide an indirect evidence that the lipid hemifusion-to-pore transition is a key event in neuroexocytosis in Drosophila. Moreover, these findings substantiate the use of LysoPC as a general agonist of membrane fusion at nerve terminals. This novel tool could contribute to the unraveling of the molecular steps involved in neuroexocytosis, particularly in Drosophila, where it is straightforward to combine it with electrophysiology and genetics.     

Enhanced Cellular Uptake of Oligonucleotides by EGF Receptor-Mediated Endocytosis in A549 Cells

Purpose. The goal of this study was to investigate the feasibility of utilizing epidermal growth factor (EGF) receptor-mediated endocytosis to enhance cellular uptake and targeting of oligonucleotides in epithelial cancer cells. To overcome the problem of endosomal entrappment associated with receptor-mediated delivery, we also examined the effects of two fusogenic peptides, polymyxin B and influenza HA2 peptide, for their capability to promote cytoplasmic delivery of oligonucleotides. Methods. A molecular conjugate consisting of EGF and poly-L-lysine (PL) was synthesized and complexed with 5′ fluorescently-labeled oligonucleotide. Cellular uptake of the complex in presence or absence of the fusogenic peptides was monitored fluorometrically. Microscopic studies were performed to visualize the intracellular distribution of the oligonucleotide. Results. Cells treated with the complex exhibited intracellular fluorescence intensity significantly enhanced over free oligonucleotide-treated controls. The uptake of the complex was shown to occur via the EGF receptor-mediated pathway. Fluorescence microscopic studies revealed cellular internalization of the complex, however, the complex appeared to be accumulated in endocytic vesicles. Exposure of the cells to complex in presence of HA2 peptide and polymyxin B resulted in a more diffused intracellular fluorescence pattern and a corresponding increase in fluorescence intensity. These results are consistent with the known fusion and destabilizing activities of the peptides. Conclusions. Since EGF receptors are overexpressed in many cancer cell types, the EGF-PL conjugate may potentially be used as an effective and selective delivery system to enhance uptake of oligonucleotides into cancer cells.