O-GlcNAc transferase promotes the nuclear localization of the focal adhesion-associated protein Zyxin to regulate UV-induced cell death.

Zyxin is a zinc-binding phosphoprotein known to regulate cell migration, adhesion, and cell survival. Zyxin also plays a role in signal transduction between focal adhesions and the nuclear compartment. However, the mechanism of Zyxin shuttling to nucleus is still unclear. Here, we identify that the GlcNAc transferase (O-linked GlcNAc [O-GlcNAc] transferase) can O-GlcNAcylate Zyxin and regulate its nuclear localization. We show that O-GlcNAc transferase O-GlcNAcylates Zyxin at two residues, serine 169 (Ser-169) and Ser-246. In addition, O-GlcNAcylation of Ser-169, but not Ser-246, enhances its interaction with 14-3-3γ, which is a phosphoserine/threonine-binding protein and is reported to bind with phosphorylated Zyxin. Furthermore, we found that 14-3-3γ could promote the nuclear localization of Zyxin after Ser-169 O-GlcNAcylation by affecting the function of the N-terminal nuclear export signal sequence; functionally, UV treatment increases the O-GlcNAcylation of Zyxin, which may enhance the nuclear location of Zyxin. Finally, Zyxin in the nucleus maintains homeodomain-interacting protein kinase 2 stability and promotes UV-induced cell death. In conclusion, we uncover that the nuclear localization of Zyxin can be regulated by its O-GlcNAcylation, and that this protein may regulate UV-induced cell death.

Novel Technology to Capture Objective Data from Patients' Recovery from Laparoscopic Endometriosis Surgery.

STUDY OBJECTIVE: To assess the feasibility of a noncontact radio sensor as an objective measurement tool to study postoperative recovery from endometriosis surgery. DESIGN: Prospective cohort pilot study. SETTING: Center for minimally invasive gynecologic surgery at an academically affiliated community hospital in conjunction with in-home monitoring. PATIENTS: Patients aged above 18 years who sleep independently and were scheduled to have laparoscopy for the diagnosis and treatment of suspected endometriosis. INTERVENTIONS: A wireless, noncontact sensor, Emerald, was installed in the subjects' home and used to capture physiologic signals without body contact. The device captured objective data about the patients' movement and sleep in their home for 5 weeks before surgery and approximately 5 weeks postoperatively. The subjects were concurrently asked to complete a daily pain assessment using a numeric rating scale and a free text survey about their daily symptoms. MEASUREMENTS AND MAIN RESULTS: Three women aged 23 years to 39 years and with mild to moderate endometriosis participated in the study. Emerald-derived sleep and wake times were contextualized and corroborated by select participant comments from retrospective surveys. In addition, self-reported pain levels and 1 sleep variable, sleep onset to deep sleep time, showed a significant (p <.01), positive correlation with next-day-pain scores in all 3 subjects: r = 0.45, 0.50, and 0.55. In other words, the longer it took the subject to go from sleep onset to deep sleep, the higher their pain score the following day. CONCLUSION: A patient's experience with pain is challenging to meaningfully quantify. This study highlights Emerald's unique ability to capture objective data in both preoperative functioning and postoperative recovery in an endometriosis population. The utility of this uniquely objective data for the clinician-patient relationship is just beginning to be explored.

Axin 1 knockdown inhibits osteoblastic apoptosis induced by Porphyromonas gingivalis lipopolysaccharide.

BACKGROUND: Porphyromonas gingivalis (Pg) is one of the pathogenic bacteria that cause periodontal diseases, lipopolysaccharide (LPS) is the key factor that triggers alveolar bone absorption. This study explored the action of Axin 1 on Pg-LPS-induced osteoblasts injury, so as to search a possible treatment for periodontal diseases. METHODS: Rat osteoblasts were dealt with Pg-LPS and Axin 1 knockdown alone or in combination. The effect of Pg-LPS and Axin 1 on osteoblast viability and apoptosis were detected by Cell Counting Kit-8 and flow cytometry. The expressions of alkaline phosphatase (ALP) and Axin 1 in processed osteoblasts were measured by western blot (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) assays. Furthermore, the role of Axin 1 knockdown in the levels of inflammatory cytokines and apoptosis-related proteins were also determined. RESULTS: Pg-LPS inhibited the viability of osteoblasts and promote apoptosis with concentration and time dependence. ALP expression in Pg-LPS-treated osteoblasts was reduced, while Axin 1 expression was increased. On the one hand, Axin 1 knockdown reversed the Pg-LPS-induced reduction of cell activity and pro-apoptosis effect. On the other hand, Axin 1 knockdown not only improved the ALP activity of Pg-LPS-treated cells, but also reduced the elevation of inflammatory cytokines (TNF-α, IL-1ß and IL-6) caused by Pg-LPS. Moreover, Pg-LPS increased the expressions of cleaved Caspase-3 and Bax, and inhibited Bcl-2 expressed, which was rescued by Axin 1 knockdown. CONCLUSION: Axin 1 knockdown inhibited Pg-LPS-induced osteoblastic apoptosis by regulating the levels of inflammatory cytokines, which may be helpful for the treatment of periodontal diseases.

Novel dental adhesive resin with crack self-healing, antimicrobial and remineralization properties.

OBJECTIVES: Secondary caries at the tooth-restoration margins is a primary reason for restoration failure. Cracks at the margins lead to leakage which can trap bacteria, producing acids to cause caries. To date, there has been no report on developing an adhesive resin that has self-healing, antibacterial and remineralizing capabilities. The objectives of this study were to: (1) develop the first self-healing adhesive with antimicrobial and remineralizing capabilities, and (2) investigate the effects of incorporating microcapsules, dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP) for the first time. METHODS: Self-healing microcapsules were synthesized with poly(urea-formaldehyde) (PUF) shells containing triethylene glycol dimethacrylate (TEGDMA) as the healing liquid. The new adhesive contained 7.5% microcapsules, 10% DMAHDM and 20% NACP. A single edge V-notched beam (SEVNB) method was used to measure the fracture toughness KIC and the autonomous crack-healing efficiency. An oral plaque microcosm biofilm model was tested. RESULTS: The new self-healing, antimicrobial and remineralizing dental adhesive matched the dentin bond strength of a commercial control (p > 0.1). The new adhesive achieved successful crack-healing, with an excellent KIC recovery of 67%. The new adhesive had strong antimicrobial activity, reducing biofilm colony-forming units by four orders of magnitude, and reducing biofilm acid production to 1/100th that of biofilms on the commercial control resin. CONCLUSIONS: A self-healing adhesive with antibacterial and remineralizing capabilities was developed for the first time. Excellent dentin bond strength, autonomous crack-healing and KIC recovery, and strong anti-biofilm properties were achieved for the new adhesive resin. CLINICAL SIGNIFICANCE: The novel method of using triple agents (self-healing microcapsules + DMAHDM + NACP) is promising for applications in dental adhesives, cements, sealants and composites to combat the two main challenges: fracture and secondary caries.

[Effect of light centrifugal force on the asporin gene expression in human periodontal ligament cells].

OBJECTIVE: To observe the expression of asporin, bone morphogenetic protein-2 (BMP-2) and alkaline phosphatase (ALP) in cultured human periodontal ligament cells in vitro under relative centrifugal force. METHODS: Human periodontal ligament cell was cultured in vitro and applied 30 ×g centrifugal force for 0, 1, 2, 6, 10 hours. The expression of asporin, BMP-2 and ALP was observed with real time-PCR. RESULTS: All the cells showed normal figuration. The expression of asporin, BMP-2, ALP in no force loading group did not have any statistical significance change (P > 0.05). In 1 hour force loading group, the expressions of asporin and BMP-2 were 0.50 ± 0.05 and 0.40 ± 0.13. In 2 hour force loading group, the expressions of asporin and BMP-2 were 0.42 ± 0.09 and 0.58 ± 0.19, which decreased significantly from no force loading group (P < 0.05). The expression of asporin and BMP-2 increased significantly in 6 hour force loading group than in 1 and 2 hour force loading groups (P < 0.05). Then the expression of asporin decreased to no force loading group level (P > 0.05) and the expression of BMP-2 decreased rapidly lower than no force loading group level (P < 0.05). During the 10 hour interval of stress loading, the expression of asporin and BMP-2 showed a positive correlation (r = 0.995, P < 0.05). CONCLUSIONS: The expression of asporin in human periodontal ligament cell was stable. Short and light centrifugal force could up-regulate the expression of asporin rapidly, and suppress the abnormal BMP-2 expression back to baseline level.

[Effect of orthodontic force on expression of Asporin after tooth auto-transplantation: an experimental study in Beagle dogs].

OBJECTIVE: To evaluate the effect of orthodontic light force on the expression of Asporin, bone morphogenetic proteins-2 (BMP-2) and alkaline phosphatase (ALP) after auto-transplantation. METHODS: Thirty-two maxillary and mandibular incisors in four 13 month-old male Beagle dogs were auto-transplanted to the other side of the same jaw. The teeth were all endodontically treated and divided into four groups, control (group 1) and three experimental groups (groups 2, 3 and 4).In control group, the teeth were unloaded. In the other three experimental groups, continuous force was applied in the 1st week (group 2), 2nd week (group 3) and 4th week (group 4) after auto-transplantation, respectively. The dogs were sacrificed in the 8th week. The mRNA expressions of Asporin,BMP-2 and ALP were examined by real time PCR. The expression of periodontal ligament associated protein-1 (PLAP-1) was examined by Western blotting. The data were statistically analyzed using SPSS 17.0 and one-way analysis of variance (ANOVA). RESULTS: In group 3, the expression of Asporin mRNA (2.047 ± 0.281) was higher than that in the other three groups, while the expression of BMP-2 (1.358 ± 0.095) was lower than that in group 2 and control group (P < 0.05). The expression of PLAP-1 (1.054 ± 0.113) in group 3 was higher than other groups, while significant difference was found between any two groups. CONCLUSIONS: Orthodontic force could stimulate the expression of Asporin and PLAP-1. The orthodontic force applied in the 2nd week after the tooth auto-transplantation, the expression of Asporin and PLAP-1 reached the highest level.