X-ray structural analyses of azide-bound cytochrome c oxidases reveal that the H-pathway is critically important for the proton-pumping activity.

Cytochrome c oxidase (CcO) is the terminal oxidase of cellular respiration, reducing O2 to water and pumping protons. X-ray structural features have suggested that CcO pumps protons via a mechanism involving electrostatic repulsions between pumping protons in the hydrogen-bond network of a proton-conducting pathway (the H-pathway) and net positive charges created upon oxidation of an iron site, heme a (Fe a2+), for reduction of O2 at another iron site, heme a3 (Fe a32+). The protons for pumping are transferred to the hydrogen-bond network from the N-side via the water channel of the H-pathway. Back-leakage of protons to the N-side is thought to be blocked by closure of the water channel. To experimentally test this, we examined X-ray structures of the azide-bound, oxidized bovine CcO and found that an azide derivative (N3--Fe a33+, CuB2+-N3-) induces a translational movement of the heme a3 plane. This was accompanied by opening of the water channel, revealing that Fe a3 and the H-pathway are tightly coupled. The channel opening in the oxidized state is likely to induce back-leakage of pumping protons, which lowers the proton level in the hydrogen-bond network during enzymatic turnover. The proton level decrease weakens the electron affinity of Fe a , if Fe a electrostatically interacts with protons in the hydrogen-bond network. The previously reported azide-induced redox-potential decrease in Fe a supports existence of the electrostatic interaction. In summary, our results indicate that the H-pathway is critical for CcO's proton-pumping function.

Impact of surface pre-reacted glass ionomer filler eluate on lipase gene expression in Candida albicans: An in vitro study.

Although a surface pre-reacted glass ionomer (S-PRG) exerts a suppressive effect on Candida albicans (C. albicans) activity and growth, its influence on the expression of the lipase gene (LIP) family including LIP1-LIP10, an indicator of clinical infection, has not yet been investigated. Therefore, in this study, we evaluated the effect of S-PRG filler eluates on LIP expression in C. albicans using real-time reverse-transcription polymerase chain reaction. Candida albicans was treated with an S-PRG filler diluted at ratios of 1:32 and 1:64 for 24 h at 37°C. The diluted S-PRG filler eluates (1:32) suppressed lipase activity in C. albicans by downregulating LIP5 (0.54±0.25 relative to that of the control) and LIP8 (0.35±0.074) expression after 24 h, which corresponded with decreased lipase activity. At a dilution factor of 1:64, there was no significant difference in LIP expression. Thus, the S-PRG filler eluate has potential to suppress fungal activity by downregulating LIP expression.

Factors Associated with Selection of Denture Adhesive Type: A Cross-Sectional Survey.

The type of denture adhesive, cream or home-liner, chosen by regular denture adhesive users and oral conditions contributing to this selection require elucidation. The factors associated with denture adhesive selection were investigated through a face-to-face survey on oral and denture conditions. Age, sex, oral moisture, masticatory performance, retention and stability of the removable denture, ridge shape, mucosal thickness, and duration of denture use were examined in cream and home-liner-type denture adhesive users who did not regularly visit a dentist. Univariate analysis and multivariate analyses were performed. There were 38 and 40 cream-type and home-liner-type adhesive users, respectively. The type of denture adhesive was significantly associated with the oral moisture value, retention, ridge shape, mucosal thickness, and duration of denture use in univariate analyses. The residual ridge conditions with large factor loadings for ridge shape and mucosal thickness and duration of denture use were significantly related to the denture adhesive selection in multivariate logistic analysis. The residual ridge conditions and duration of denture use were significant factors in the selection of cream- and home-liner-type denture adhesives. These results can provide appropriate guidance based on the adhesives patients without dental supervision are more likely to choose.

Impact of direct restorative dental materials on surface root caries treatment. Evidence based and current materials development: A systematic review.

This systematic review provides an update on the development and efficacy of direct restorative dental materials for root caries interventions from in vitro and clinical studies. PubMed, Embase, and Web of Science were searched using specific MeSH keywords. Full articles from September 1990 to October 2021 were collected. Additional articles were identified by reference retrieval and manual searching. Studies not related to restorative materials for root caries treatment, case reports, non-original articles, and/or articles not written in English were excluded. Bias risk assessment was performed for the clinical studies. Forty-two articles (eleven clinical studies and thirty-one in vitro studies) were included for analysis. Most in vitro studies indicated an excellent cariostatic effect of glass ionomer cement. Resin-modified glass ionomer restorations also presented reduced recurrent caries activity but had a lower efficacy than glass ionomer cement restorations. For composite resin restorations, the main material development strategies are to strengthen the tooth structure and integrate antimicrobial activity. The clinical studies offered limited data, so the most appropriate material for surface root caries treatment is still inconclusive. However, atraumatic restorative treatment (ART) is an alternative treatment for patients with limiting conditions. Further clinical studies are required to confirm the efficacy of bioactive materials.

Tissue Conditioner Incorporating a Nano-Sized Surface Pre-Reacted Glass-Ionomer (S-PRG) Filler.

We aimed to evaluate the properties of a novel tissue conditioner containing a surface pre-reacted glass-ionomer (S-PRG) nanofiller. Tissue conditioners containing 0 (control), 2.5, 5, 10, 20, or 30 wt% S-PRG nanofiller or 10 or 20 wt% S-PRG microfiller were prepared. The S-PRG nanofillers and microfillers were observed using scanning electron microscopy. The ion release, acid buffering capacity, detail reproduction, consistency, Shore A0 hardness, surface roughness, and Candida albicans adhesion of the tissue conditioners were examined. The results indicated that the nanofiller particles were smaller and more homogeneous in size than the microfiller particles. In addition, Al, B, F, and Sr ions eluted from S-PRG were generally found to decrease after 1 day. Acid neutralization was confirmed in a concentration-dependent manner. The mechanical properties of tissue conditioners containing S-PRG nanofiller were clinically acceptable according to ISO standard 10139-1:2018, although the surface roughness increased with increasing filler content. Conditioners with 5-30 wt% nanofiller had a sublethal effect on C. albicans and reduced fungal adhesion in vitro. In summary, tissue conditioner containing at least 5 wt% S-PRG nanofiller can reduce C. albicans adhesion and has potential as an alternative soft lining material.

Novel antimicrobial denture adhesive containing S-PRG filler.

The antimicrobial effects of denture adhesives containing novel surface pre-reacted glass-ionomer (S-PRG) fillers were assessed. We prepared denture adhesives containing S-PRG (particle sizes: 1 and 3 µm; quantities: 5, 7.5, and 10 wt%). We evaluated acid buffering capacity, ion release, and antimicrobial effects of denture adhesives with and without S-PRG. Significantly higher pH changes were observed in 1 µm S-PRG adhesives than in 3 µm S-PRG adhesives. Adhesives containing 7.5 and 10 wt% S-PRG exhibited significantly higher ion release than adhesives with 5 wt% S-PRG. The 1µm-10wt% S-PRG denture adhesive exhibited significantly lower colony-forming units on the denture adhesive contact surface than in the control group; additionally, it exhibited excellent acid buffering capacity, ion release properties, and antimicrobial effect against C. albicans, C. glabrata, S. mutans, and A. naeslundii. Longer contact periods resulted in significantly lower adhesion of Candida albicans to the denture base resin treated with denture adhesive.

A nanosecond time-resolved XFEL analysis of structural changes associated with CO release from cytochrome c oxidase.

Bovine cytochrome c oxidase (CcO), a 420-kDa membrane protein, pumps protons using electrostatic repulsion between protons transferred through a water channel and net positive charges created by oxidation of heme a (Fe a ) for reduction of O2 at heme a3 (Fe a3). For this process to function properly, timing is essential: The channel must be closed after collection of the protons to be pumped and before Fe a oxidation. If the channel were to remain open, spontaneous backflow of the collected protons would occur. For elucidation of the channel closure mechanism, the opening of the channel, which occurs upon release of CO from CcO, is investigated by newly developed time-resolved x-ray free-electron laser and infrared techniques with nanosecond time resolution. The opening process indicates that CuB senses completion of proton collection and binds O2 before binding to Fe a3 to close the water channel using a conformational relay system, which includes CuB, heme a3, and a transmembrane helix, to block backflow of the collected protons.