Double base mismatches mediated catalytic hairpin assembly for enzyme-free single-base mutation detection: integrating signal recognition and amplification in one.

Single nucleotide polymorphism (SNP) biosensors are emerging rapidly for their promising applications in human disease prevention diagnosis, treatment, and prognosis. However, it remains a bottleneck in equipping simple and stable biosensors with the traits of high sensitivity, non-enzyme, and low cost. Double base mismatches mediated chain displacement reactions have attracted fascinating advantages of tailorable thermodynamics stability, non-enzyme, and excellent assembly compliance to involvement in SNP identification. As the base mismatch position and amount in DNA sequence can be artificially adjusted, it provides plenty of selectivity and specificity for exploring perfect biosensors. Herein, a biosensor with double base mismatches mediated catalytic hairpin assembly (CHA) is designed via one base mismatch in the toehold domain and the other base mismatch in the stem sequence of hairpin 1 (H1) by triggering CHA reaction to achieve selective amplification of the mutation target (MT) and fluorescence resonance energy transfer (FRET) effect that is composed of Cy3 and Cy5 terminally attached H1 and hairpin 2 (H2). Depending on the rationally designed base mismatch position and toehold length, the fabricated biosensors show superior SNP detection performance, exhibiting a good linearity with high sensitivity of 6.6 fM detection limit and a broad detection abundance of 1%. The proposed biosensor can be used to detect the KRAS mutation gene in real samples and obtain good recoveries between 106 and 116.99%. Remarkably, these extendible designs of base mismatches can be used for more types of SNP detection, providing flexible adjustment based on base mismatch position and toehold length variations, especially for their thermodynamic model for DNA-strand displacement reactions.

Immediate loaded fixed complete dentures supported by implants in patients with a history of periodontitis: A retrospective cohort study of 2 to 7 years.

STATEMENT OF PROBLEM: The outcome of implant-supported fixed complete dentures in edentulous patients with a history of periodontitis is unclear. PURPOSE: The purpose of this retrospective clinical study was to assess the clinical outcomes of immediate loaded fixed complete dentures in individuals with a history of periodontitis and to analyze risk factors related to implant failure. MATERIAL AND METHODS: A total of 642 implants (146 prostheses) in 119 patients were included. The follow-up period ranged from 2 to 7 years. Implant survival rates, marginal bone loss, mechanical complications, biologic complications, and patient satisfaction were evaluated. The Pearson chi-square test, independent samples t test, and multivariate generalized estimating equation were performed for statistical analysis (α=.05). RESULTS: Eleven implants in 9 patients failed, leading to overall survival rates of 98.3% at the implant level and 92.4% at the patient level. The mean ±standard deviation marginal bone loss was 0.62 ±0.86 mm, and marginal bone loss did not differ significantly between axial and tilted implants (P>.05). Mechanical complications were detected in 55 (37.7%) definitive prostheses; biologic complications were detected in 318 (49.5%) implants. Smokers had a significantly lower survival rate than nonsmokers (odds ratio: 6.880, P=.013). Bruxers had a significantly higher incidence of mechanical complications than nonbruxers (P<.001). CONCLUSIONS: The immediate loaded fixed complete denture supported by implants is a suitable treatment option for edentulous patients with a history of periodontitis, with high survival implant rates. Smoking is a risk factor for implant failure. Bruxism may increase the incidence of mechanical complications with implant-supported fixed complete dentures, and the overall biologic complication incidence is comparatively high.

Cleaning effects of decontamination methods on clinically failed TiUnite implants and their impacts on surface roughness and chemistry: An in vitro pilot study.

PURPOSE: To evaluate the in vitro cleaning effects of different decontamination methods and their impacts on surface characteristics using clinically failed TiUnite implants (Nobel Biocare, Kloten, Switzerland). MATERIALS AND METHODS: Thirty clinically failed TiUnite implants were treated using different decontamination methods. Group 1 (control group) received physiological saline irrigation; Group 2 underwent erythritol powder air polishing (AIRFLOW Master Piezon, EMS Dental, Nyon, Switzerland); Group 3 was treated with erythritol powder air polishing with ethylenediaminetetraacetic acid brushing (FileRite PRC, Pulpdent, Watertown, MA, USA); Group 4 received ultrasonic scaling with polyetheretherketone tips (EMS Dental); Group 5 underwent ultrasonic scaling with polyetheretherketone tips with ethylenediaminetetraacetic acid; and Group 6 was treated with a combination of ultrasonic scaling with polyetheretherketone tips, erythritol powder air polishing and ethylenediaminetetraacetic acid. Surface cleaning effects, quantified by relative contaminated area reduction and visual analogue scale score, as well as surface roughness and chemistry, were assessed after decontamination. The cleaning effects of each decontamination method were also compared between TiUnite and SLA (sandblasted, large-grit acid-etched; Straumann, Basel, Switzerland) implants. RESULTS: Group 6 showed the highest relative contaminated area reduction (stereoscopic microscopy 83.92%, scanning electron microscopy 96.40%), visual analogue scale score (2.83) and reduction in surface roughness (thread bottom -0.78 µm, tip -1.35 µm), as well as an almost maximal decrease in the proportion of carbon (thread bottom -12.33%, tip -8.77%) and increase in that of titanium (thread bottom 13.71%, tip 10.73%). Polyetheretherketone remnants were observed in Groups 4 and 5 but appeared to be reduced in Group 6. When comparing the outcomes with those for SLA implants, no significant differences were found. CONCLUSION: Within the limitations of the present study, the combination of ultrasonic scaling with polyetheretherketone tips, erythritol powder air polishing and ethylenediaminetetraacetic brushing achieved reasonable cleaning effects. The original surface modification did not seem to have any impact on the decontamination results for any of the methods examined.

IL-1 Receptor Antagonist Protects the Osteogenesis Capability of Gingival-Derived Stem/Progenitor Cells under Inflammatory Microenvironment Induced by Porphyromonas gingivalis Lipopolysaccharides.

Mesenchymal stem cells (MSCs) have been considered to be a future treatment option for periodontitis due to their excellent regenerative capability. However, it is still a challenge to protect MSCs' biological properties from multiple bacterial toxins in local inflammatory environment. The present study is aimed at investigating the treatment effect of interleukin-1 receptor antagonist (IL-1ra) on cell proliferation, migration, and osteogenic differentiation of gingival-derived mesenchymal stem cells (GMSCs) under an inflammatory microenvironment induced by Porphyromonas gingivalis lipopolysaccharides (P. gingivalis-LPS). GMSCs derived from Sprague-Dawley (SD) rats' free gingival tissues were treated with P. gingivalis-LPS (10 µg/mL) to create in vitro inflammatory environment. Different concentrations of IL-1ra (0.01-1 µg/mL) were used to antagonize the negative effect of LPS. Cell behaviors including proliferation, cloning formation unit (CFU), cell migration, osteogenic differentiation, mineral deposition, and cytokine production were assessed to investigate the protection effect of IL-1ra on GMSCs under inflammation. The toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) pathway activated by LPS was evaluated by real-time quantitative polymerase chain reaction (RT-PCR) and western blot. In response to P. gingivalis-LPS treatment, cell numbers, cloning formation rate, cell migration rate, proinflammatory cytokine production, and osteogenic differentiation-associated protein/mRNA expressions as well as mineralized nodules were suppressed in a time-dependent manner. These negative effects were effectively attenuated by IL-1ra administration in a time- and dose-dependent manner. In addition, mRNA expressions of TLR4 and IkBα decreased dramatically when IL-1ra was added into LPS-induced medium. IL-1ra also reversed the LPS-induced TLR4/NF-κB activation as indicated by western blot. The present study revealed that IL-1ra decreased inflammatory cytokine production in a supernatant, so as to protect GMSCs' osteogenesis capacity and other biological properties under P. gingivalis-LPS-induced inflammatory environment. This might be explained by IL-1ra downregulating TLR4-mediated NF-κB signaling pathway activation.

Oil degradation and biosurfactant production by the deep sea bacterium Dietzia maris As-13-3.

Recent investigations of extreme environments have revealed numerous bioactive natural products. However, biosurfactant-producing strains from deep sea extreme environment are largely unknown. Here, we show that Dietzia maris As-13-3 isolated from deep sea hydrothermal field could produce di-rhamnolipid as biosurfactant. The critical micelle concentration (CMC) of the purified di-rhamnolipid was determined to be 120 mgL(-1), and it lowered the surface tension of water from 74 ± 0.2 to 38 ± 0.2 mN m(-1). Further, the alkane metabolic pathway-related genes and di-rhamnolipid biosynthesis-related genes were also analyzed by the sequencing genome of D. maris As-13-3 and quantitative real-time PCR (Q-PCR), respectively. Q-PCR analysis showed that all these genes were induced by n-Tetradecane, n-Hexadecane, and pristane. To the best of our knowledge, this is first report about the complete pathway of the di-rhamnolipid synthesis process in the genus Dietzia. Thus, our study provided the insights into Dietzia in respects of oil degradation and biosurfactant production, and will help to evaluate the potential of Dietzia in marine oil removal.