The PKA signaling pathway regulates the association of the autophagy initiation complex with the lipidation machinery.

A key step in autophagy is the conjugation by the E3-like Atg12-Atg5-Atg16 complex of the ubiquitin-like protein Atg8 to phosphatidylethanolamine on the autophagosomal membrane, a process known as lipidation. Previous work in yeast showed that recruitment of the E3-like complex to the preautophagosomal structure is mediated by the interaction of Atg16 with the phosphatidylinositol 3-phosphate-binding protein Atg21, and by the association of Atg12 with the scaffold protein of the Atg1 kinase complex, Atg17. Here, we conducted a reverse two-hybrid screen to identify residues in Atg17 and Atg12 critical for Atg17-Atg12 binding, and used these data to generate a docking model of Atg12-Atg5-Atg16 with the Atg17 complex.

Virtual Reality as Active Distraction in Laceration Repair: A Game Changer?

Objectives: We conducted an unblinded, randomized control trial to determine if immersive virtual reality (VR) goggles decrease pain and fear scores in children undergoing laceration repair in the pediatric emergency department (PED) compared to the standard of care. Secondary outcomes included duration of procedure, physical holding, anxiolytic usage, and desire to use VR goggles again.

Methods: Ninety-one patients aged 6-17 years in a PED with simple lacerations sutured by PED staff completed surveys.

Methods to Assess Autophagic Activity in Dictyostelium.

Autophagy is an intracellular clearance and recycling pathway that delivers different types of cargos to lysosomes for degradation. In recent years, autophagy has attracted considerable medical interest, and many different techniques are being developed to study this process in experimental models such as Dictyostelium. Here we describe the use of different autophagic markers in confocal microscopy, in vivo and also in fixed cells.

Coiled-coil-mediated dimerization of Atg16 is required for binding to the PROPPIN Atg21.

PROPPINs/WIPIs are β-propeller proteins that bind phosphoinositides and contribute to the recruitment of protein complexes involved in membrane remodelling processes such as autophagosome formation and endosomal trafficking. Yeast Atg21 and mammalian WIPI2 interact with Atg16/ATG16L1 to mediate recruitment of the lipidation machinery to the autophagosomal membrane. Here, we used the reverse double two-hybrid method (RD2H) to identify residues in Atg21 and Atg16 critical for protein-protein binding.

Kinetics of Nisin-Induced Pore Formation in Giant Unilamellar Vesicles.

We have studied the kinetics of pore formation in giant unilamellar vesicles (GUV) with the antimicrobial peptide nisin. The role of charged lipid composition in the rate of pore formation by nisin in the vesicle membrane is investigated using fluorescence microscopy. We propose a model and obtain an analytical expression for the variation in the fluorescence intensity of a GUV as a function of time.