ULK1 (unc51-like autophagy activating kinase 1) is a serine/threonine kinase that plays a key role in regulating macroautophagy/autophagy induction in response to amino acid starvation. Despite the recent progress in understanding ULK1 functions, the molecular mechanism by which ULK1 regulates the induction of autophagy remains elusive. In this study, we determined that ULK1 phosphorylates Ser30 of BECN1 (Beclin 1) in association with ATG14 (autophagy-related 14) but not with UVRAG (UV radiation resistance associated). The Ser30 phosphorylation was induced by deprivation of amino acids or treatments with Torin 1 or rapamycin, the conditions that inhibit MTORC1 (mechanistic target of rapamycin complex 1), and requires ATG13 and RB1CC1 (RB1 inducible coiled-coil 1), proteins that interact with ULK1. Hypoxia or glutamine deprivation, which inhibit MTORC1, was also able to increase the phosphorylation in a manner dependent upon ULK1 and ULK2. Blocking the BECN1 phosphorylation by replacing Ser30 with alanine suppressed the amino acid starvation-induced activation of the ATG14-containing PIK3C3/VPS34 (phosphatidylinositol 3-kinase catalytic subunit type 3) kinase, and reduced autophagy flux and the formation of phagophores and autophagosomes. The Ser30-to-Ala mutation did not affect the ULK1-mediated phosphorylations of BECN1 Ser15 or ATG14 Ser29, indicating that the BECN1 Ser30 phosphorylation might regulate autophagy independently of those 2 sites. Taken together, these results demonstrate that BECN1 Ser30 is a ULK1 target site whose phosphorylation activates the ATG14-containing PIK3C3 complex and stimulates autophagosome formation in response to amino acid starvation, hypoxia, and MTORC1 inhibition.

Anh JW, et al. Korean J Orthod. 2016 Jan;46(1):3-12. http://dx.doi.org/10.4041/kjod.2016.46.1.3

The clinical application of 3D-printed provisional restorations is increasing due to expansion of intraoral scanners, easy dental computer-aided design (CAD) software, and improved 3D printing speed. This study compared flexural strength of 3D-printed three-unit fixed dental prostheses with that of conventionally fabricated and milled restorations. A metal jig of two abutments and pontic space and an indenter for flexural strength measurement were fabricated. A three-unit fixed dental prosthesis was designed and manufactured using three additive manufacturing technologies, with subtractive manufacturing and a conventional method as controls. Digital light processing (DLP) group specimens were prepared from a polymethyl methacrylate (PMMA)-based resin and printed with a DLP printer. Stereolithography (SLA) group specimens were prepared from PMMA-based resin and printed with an SLA printer, and fused deposition modeling (FDM) group specimens were from a polylactic acid-based resin and printed with an FDM printer. Flexural strength was investigated using a universal testing machine, and the results were statistically analyzed. DLP and SLA groups had significantly higher flexural strength than the conventional group (p < 0.001). No significant difference was observed in flexural strength between DLP and SLA groups. The FDM group showed only dents but no fracture. The results of this study suggest that provisional restorations fabricated by DLP and SLA technologies provide adequate flexural strength for dental use.