Objective

TMEM55B (transmembrane protein 55B) is a phosphatidylinositol-(4,5)-bisphosphate (PI[4,5]P₂) phosphatase that regulates cellular cholesterol, modulates LDLR (low-density lipoprotein receptor) decay, and lysosome function. We tested the effects of Tmem55b knockdown on plasma lipids in mice and assessed the roles of LDLR lysosomal degradation and change in (PI[4,5]P₂) in mediating these effects. Approach and Results: Western diet-fed C57BL/6J mice were treated with antisense oligonucleotides against Tmem55b or a nontargeting control for 3 to 4 weeks. Hepatic Tmem55b transcript and protein levels were reduced by ≈70%, and plasma non-HDL (high-density lipoprotein) cholesterol was increased ≈1.8-fold (P<0.0001). Immunoblot analysis of fast protein liquid chromatography (FPLC) fractions revealed enrichment of ApoE-containing particles in the LDL size range. In contrast, Tmem55b knockdown had no effect on plasma cholesterol in Ldlr^-/- mice. In primary hepatocytes and liver tissues from Tmem55b knockdown mice, there was decreased LDLR protein. In the hepatocytes, there was increased lysosome staining and increased LDLR-lysosome colocalization. Impairment of lysosome function (incubation with NH₄Cl or knockdown of the lysosomal proteins LAMP1 or RAB7) abolished the effect of TMEM55B knockdown on LDLR in HepG2 (human hepatoma) cells. Colocalization of the recycling endosome marker RAB11 (Ras-related protein 11) with LDLR in HepG2 cells was reduced by 50% upon TMEM55B knockdown. Finally, knockdown increased hepatic PI(4,5)P₂ levels in vivo and in HepG2 cells, while TMEM55B overexpression in vitro decreased PI(4,5)P₂. TMEM55B knockdown decreased, whereas overexpression increased, LDL uptake in HepG2 cells. Notably, the TMEM55B overexpression effect was reversed by incubation with PI(4,5)P_2. Conclusions: These findings indicate a role for TMEM55B in regulating plasma cholesterol levels by affecting PI(4,5)P₂-mediated LDLR lysosomal degradation.

PREdiction of NOn-LINear soil behavior (PRENOLIN) is an international benchmark aiming to test multiple numerical simulation codes that are capable of predicting nonlinear seismic site response with various constitutive models. One of the objectives of this project is the assessment of the uncertainties associated with nonlinear simulation of 1D site effects. A first verification phase (i.e., comparison between numerical codes on simple idealistic cases) will be followed by a validation phase, comparing the predictions of such numerical estimations with actual strongmotion recordings obtained at well-known sites. The benchmark presently involves 21 teams and 23 different computational codes. We present here the main results of the verification phase dealing with simple cases. Three different idealized soil profiles were tested over a wide range of shear strains with different input motions and different boundary conditions at the sediment/bedrock interface. A first iteration focusing on the elastic and viscoelastic cases was proved to be useful to ensure a common understanding and to identify numerical issues before pursuing the nonlinear modeling. Besides minor mistakes in the implementation of input parameters and output units, the initial discrepancies between the numerical results can be attributed to (1) different understanding of the expression “input motion” in different communities, and (2) different implementations of material damping and possible numerical energy dissipation. The second round of computations thus allowed a convergence of all teams to the Haskell–Thomson analytical solution in elastic and viscoelastic cases. For nonlinear computations, we investigate the epistemic uncertainties related only to wave propagation modeling using different nonlinear constitutive models. Such epistemic uncertainties are shown to increase with the strain level and to reach values around 0.2 (log10 scale) for a peak ground acceleration of 5 m=s2 at the base of the soil column, which may be reduced by almost 50% when the various constitutive models used the same shear strength and damping implementation.