Strength of titanium-zirconium alloy implants with a conical connection after implantoplasty.

STATEMENT OF PROBLEM: Peri-implantitis occurs around dental implants, and implantoplasty has been used to address this ongoing disease; however, the changes to the physical properties of an implant after implantoplasty have not been well documented. PURPOSE: The purpose of this in vitro study was to determine the effect of implantoplasty on fracture strength and the load required for plastic deformation after cyclic fatigue on dental implants. MATERIAL AND METHODS: Twenty-six titanium/zirconium (TiZr) alloy implants (Roxolid Bone Level Implant; 4.1×10 mm) were embedded with 50% thread exposure and divided into 4 groups based on whether they had implantoplasty treatment by using different diamond rotary instruments and/or cyclic loading at 250 N for 2 million cycles: C0 (control, no cyclic loading), T0 (test, no cyclic loading), CM (control, cyclic loading), and TM (test, cyclic loading). After implantoplasty and/or cyclic loading, all implants underwent a load-to-failure test. The maximum fracture strength (FS) and load required for the onset of plastic deformation (PD) were recorded in Newtons. One-way ANOVA and nonparametric comparisons with control by using the Dunn and Wilcoxon method for joint ranking were used for statistical analysis. RESULTS: The mean ±standard deviation FS for C0, CM, T0, and TM was 1465.2 ±86.4 N, 1480.7 ±64.1 N, 1299.3 ±123.8 N, and 1252.1 ±85.7 N, respectively. The mean ±standard deviation load for onset of PD for C0, CM, T0, and TM was 860.2 ±88.1 N, 797.0 ±130.5 N, 776.5 ±181.8 N, and 631.3 ±84.5 N, respectively. The TM group had a significantly lower FS and PD than the C0, CM, and T0 groups (P<.05) CONCLUSIONS: Both fracture strength (FS) and the onset of plastic deformation (PD) were significantly reduced after a TiZr alloy implant received implantoplasty and cyclic loading.

PITPs as targets for selectively interfering with phosphoinositide signaling in cells.

Sec14-like phosphatidylinositol transfer proteins (PITPs) integrate diverse territories of intracellular lipid metabolism with stimulated phosphatidylinositol-4-phosphate production and are discriminating portals for interrogating phosphoinositide signaling. Yet, neither Sec14-like PITPs nor PITPs in general have been exploited as targets for chemical inhibition for such purposes. Herein, we validate what is to our knowledge the first small-molecule inhibitors (SMIs) of the yeast PITP Sec14. These SMIs are nitrophenyl(4-(2-methoxyphenyl)piperazin-1-yl)methanones (NPPMs) and are effective inhibitors in vitro and in vivo. We further establish that Sec14 is the sole essential NPPM target in yeast and that NPPMs exhibit exquisite targeting specificities for Sec14 (relative to related Sec14-like PITPs), propose a mechanism for how NPPMs exert their inhibitory effects and demonstrate that NPPMs exhibit exquisite pathway selectivity in inhibiting phosphoinositide signaling in cells. These data deliver proof of concept that PITP-directed SMIs offer new and generally applicable avenues for intervening with phosphoinositide signaling pathways with selectivities superior to those afforded by contemporary lipid kinase-directed strategies.

Zebrafish class 1 phosphatidylinositol transfer proteins: PITPbeta and double cone cell outer segment integrity in retina.

Phosphatidylinositol transfer proteins (PITPs) in yeast co-ordinate lipid metabolism with the activities of specific membrane trafficking pathways. The structurally unrelated metazoan PITPs (mPITPs), on the other hand, are an under-investigated class of proteins. It remains unclear what biological activities mPITPs discharge, and the mechanisms by which these proteins function are also not understood. The soluble class 1 mPITPs include the PITPalpha and PITPbeta isoforms. Of these, the beta-isoforms are particularly poorly characterized. Herein, we report the use of zebrafish as a model vertebrate for the study of class 1 mPITP biological function. Zebrafish express PITPalpha and PITPbeta-isoforms (Pitpna and Pitpnb, respectively) and a novel PITPbeta-like isoform (Pitpng). Pitpnb expression is particularly robust in double cone cells of the zebrafish retina. Morpholino-mediated protein knockdown experiments demonstrate Pitpnb activity is primarily required for biogenesis/maintenance of the double cone photoreceptor cell outer segments in the developing retina. By contrast, Pitpna activity is essential for successful navigation of early developmental programs. This study reports the initial description of the zebrafish class 1 mPITP family, and the first analysis of PITPbeta function in a vertebrate.

Resurrection of a functional phosphatidylinositol transfer protein from a pseudo-Sec14 scaffold by directed evolution.

Sec14-superfamily proteins integrate the lipid metabolome with phosphoinositide synthesis and signaling via primed presentation of phosphatidylinositol (PtdIns) to PtdIns kinases. Sec14 action as a PtdIns-presentation scaffold requires heterotypic exchange of phosphatidylcholine (PtdCho) for PtdIns, or vice versa, in a poorly understood progression of regulated conformational transitions. We identify mutations that confer Sec14-like activities to a functionally inert pseudo-Sec14 (Sfh1), which seemingly conserves all of the structural requirements for Sec14 function. Unexpectedly, the "activation" phenotype results from alteration of residues conserved between Sfh1 and Sec14. Using biochemical and biophysical, structural, and computational approaches, we find the activation mechanism reconfigures atomic interactions between amino acid side chains and internal water in an unusual hydrophilic microenvironment within the hydrophobic Sfh1 ligand-binding cavity. These altered dynamics reconstitute a functional "gating module" that propagates conformational energy from within the hydrophobic pocket to the helical unit that gates pocket access. The net effect is enhanced rates of phospholipid-cycling into and out of the Sfh1* hydrophobic pocket. Taken together, the directed evolution approach reveals an unexpectedly flexible functional engineering of a Sec14-like PtdIns transfer protein-an engineering invisible to standard bioinformatic, crystallographic, and rational mutagenesis approaches.