Nek4 regulates mitochondrial respiration and morphology.

Nek4 is a serine/threonine kinase which has been implicated in primary cilia stabilization, DNA damage response, autophagy and epithelial-to-mesenchymal transition. The role of Nek4 in cancer cell survival and chemotherapy resistance has also been shown. However, the precise mechanisms by which Nek4 operates remain to be elucidated. Here, we show that Nek4 overexpression activates mitochondrial respiration coupled to ATP production, which is paralleled by increased mitochondrial membrane potential, and resistance to mitochondrial DNA damage. Congruently, Nek4 depletion reduced mitochondrial respiration and mtDNA integrity. Nek4 deficiency caused mitochondrial elongation, probably via reduced activity of the fission protein DRP1. In Nek4 overexpressing cells, the increase in mitochondrial fission was concomitant to enhanced phosphorylation of DRP1 and Erk1/2 proteins, and the effects on mitochondrial respiration were abolished in the presence of a DRP1 inhibitor. This study shows Nek4 as a novel regulator of mitochondrial function that may explain the joint appearance of high mitochondrial respiration and mitochondrial fragmentation.

On Broken Ne(c)ks and Broken DNA: The Role of Human NEKs in the DNA Damage Response.

NIMA-related kinases, or NEKs, are a family of Ser/Thr protein kinases involved in cell cycle and mitosis, centrosome disjunction, primary cilia functions, and DNA damage responses among other biological functional contexts in vertebrate cells. In human cells, there are 11 members, termed NEK1 to 11, and the research has mainly focused on exploring the more predominant roles of NEKs in mitosis regulation and cell cycle. A possible important role of NEKs in DNA damage response (DDR) first emerged for NEK1, but recent studies for most NEKs showed participation in DDR. A detailed analysis of the protein interactions, phosphorylation events, and studies of functional aspects of NEKs from the literature led us to propose a more general role of NEKs in DDR. In this review, we express that NEK1 is an activator of ataxia telangiectasia and Rad3-related (ATR), and its activation results in cell cycle arrest, guaranteeing DNA repair while activating specific repair pathways such as homology repair (HR) and DNA double-strand break (DSB) repair. For NEK2, 6, 8, 9, and 11, we found a role downstream of ATR and ataxia telangiectasia mutated (ATM) that results in cell cycle arrest, but details of possible activated repair pathways are still being investigated. NEK4 shows a connection to the regulation of the nonhomologous end-joining (NHEJ) repair of DNA DSBs, through recruitment of DNA-PK to DNA damage foci. NEK5 interacts with topoisomerase IIß, and its knockdown results in the accumulation of damaged DNA. NEK7 has a regulatory role in the detection of oxidative damage to telomeric DNA. Finally, NEK10 has recently been shown to phosphorylate p53 at Y327, promoting cell cycle arrest after exposure to DNA damaging agents. In summary, this review highlights important discoveries of the ever-growing involvement of NEK kinases in the DDR pathways. A better understanding of these roles may open new diagnostic possibilities or pharmaceutical interventions regarding the chemo-sensitizing inhibition of NEKs in various forms of cancer and other diseases.

NEK5 interacts with LonP1 and its kinase activity is essential for the regulation of mitochondrial functions and mtDNA maintenance.

Little is known about Nima-related kinase (NEKs), a widely conserved family of kinases that have key roles in cell-cycle progression. Nevertheless, it is now clear that multiple NEK family members act in networks, not only to regulate specific events of mitosis, but also to regulate metabolic events independently of the cell cycle. NEK5 was shown to act in centrosome disjunction, caspase-3 regulation, myogenesis, and mitochondrial respiration. Here, we demonstrate that NEK5 interacts with LonP1, an AAA+ mitochondrial protease implicated in protein quality control and mtDNA remodeling, within the mitochondria and it might be involved in the LonP1-TFAM signaling module. Moreover, we demonstrate that NEK5 kinase activity is required for maintaining mitochondrial mass and functionality and mtDNA integrity after oxidative damage. Taken together, these results show a new role of NEK5 in the regulation of mitochondrial homeostasis and mtDNA maintenance, possibly due to its interaction with key mitochondrial proteins, such as LonP1.

Color stability of dental enamel bleached with violet LED associated with or without Low concentration peroxide gels.

BACKGROUND: This study aimed to evaluate the in vitro color of dental enamel bleached with the violet LED, associated with or without low-concentration of peroxide; analyzed in two time intervals: 24 h later and 6 months after the treatment. METHODS: Ninety fragments of bovine teeth (6 × 6 mm and 3 mm thick) were randomly divided into 6 groups according to bleaching treatment: NB - no bleaching, VL - Violet LED, HP- 7.5 % hydrogen peroxide, HP + VL - 7.5 % hydrogen peroxide + violet LED, CP - 22 % carbamide peroxide, CP + VL - 22 % carbamide peroxide + violet LED. The color change was analyzed by using a spectrophotometer, at time intervals of 24 h and 6 months after performing the bleaching techniques (n = 12). Scanning electron microscopy (SEM) was performed to verify the enamel surface morphology after treatment (n = 3). RESULTS: The color change data were analyzed using ANOVA and a post-hoc Tukey tests (α = 0.05). The VL group showed chromatic changes after 24 h of treatment, however the groups submitted to bleaching gel treatments associated with or without violet LED (CP + VL, CP, HP, HP + VL) showed the highest color change values at all time intervals analyzed, with color stability after 6 months of treatment for the CP + VL group. Scanning electron microscopy analysis showed the greatest change in enamel surface for Groups CP and HP. CONCLUSIONS: It could be concluded that violet LED had immediate bleaching effect without promoting significant changes in enamel morphology, however the association with carbamide peroxide 22 % showed color stability and greater bleaching efficacy than the use of violet LED alone, after 6 months.

Cross-infection control in clinical training: Development of a device for storing a composite resin in restorative dentistry.

This study presented a biosafety device for the hygienic storage of composite resin during restorative procedures in teaching clinics, RESTAURASAFE, and to compare the color stability, surface hardness, and degree of conversion of the composite resin made with the proposed device to those of conventionally made resins. Esthet-X HD composite was manipulated directly from the syringe, fractionated doses, dappen dish or RESTAURASAFE. Disc-shaped specimens were immersed in artificial saliva and coffee for 60 days and the color stability and the Vickers surface hardness were evaluated during 60 days. Degree of conversion was assessed after immersing specimens in saliva (baseline). The surface morphology analysis of composite resin has taken by scanning electron microscopy (SEM). The data were evaluated statistically by repeated measures ANOVA (α=.05). No difference of color stability was observed for saliva and coffee regardless of storing devices. The resin manipulated directly from the syringe presented lower hardness, and fractionated doses presented lower degree of conversion than the RESTAURASAFE. SEM analysis showed that after artificial aging in coffee for 60 days, the specimens prepared with fractionated doses of resin displayed the fewest changes in surface morphology, followed by those prepared with the RESTAURASAFE. We conclude that RESTAURASAFE allows to obtain similar color stability, higher hardness, and degree of conversion of the composite in comparison to the other storing devices.

Evaluation of masticatory capacity and efficiency in total and partial edentulous patients in Araraquara

Aim: This study aims to compare masticatory performance between total prosthesis users and individuals with partial or total natural dentition to evaluate masticatory capacity and efficiency in individuals in Araraquara/SP. Material and Methods: 31 individuals between 55 and 99 years old were evaluated, both genders, frequent users of Dentistry Clinic of the University of Araraquara (Uniara) and residents at Recanto Feliz, in the same town. A cognitive evaluation was carried out through a mini mental state examination in order to define inclusion criteria to individuals of sample. Next, a questionnaire was applied, with pre-coded questions on the masticatory capacity and an efficiency test, through the grinding test, where patients received 5 almonds to be grinded and not swallowed. The fragments were poured in granulometric sieves (2mm, 4mm and 4,75mm) to analyze the food fragmentation under tap water. Participants were divided into 3 groups: G1 (n=13) ­ superior and inferior total prosthesis user, or total or partial edentulous; G2 (n=4) ­ Total prosthesis user and/or partial removal prosthesis user (PRP) or partial dentition; and G3 (n=14) ­ complete or partial dentition or PRP user. Results: Efficiency test results showed the G1 patients presented good efficiency and 8 were considered poor; G2 patients were all classified as poor, and G3 patients were 9 poor, 2 regular and 3 good. Regarding to the questionnaires, most patients from the 3 groups felt impossibility to masticate some food, and need special preparation to chew. On take more time to masticate when compared to other people, all patients from G2 answered affirmatively, while only 46.1% G1 and 50% G3 gave the same answer.Just 15.4% G1 and 28.6% G2 are not satisfied with the tame they take.About avoid in front of other people, most patients from G2 and G3 use to, however, 69.2% G1 answered negatively. Regarding to the masticatory capacity self-evaluation, they answered regular. Conclusions: Masticatory efficiency of people using removable prosthesis did not achieve the ideal of a complete dentition. However, it is still better than in individuals with partial dentition or not rehabilitated.

Roughness and morphology of composites: influence of type of material, fluoride solution, and time.

This study evaluated the effect of fluoride solutions on surface roughness and morphology of composites in the short and long term. Specimens were randomly assigned to experimental groups (n=5) according to type of composite (nanofilled, microhybrid, microfilled) and immersion media (artificial saliva, 0.05% sodium fluoride solution, Fluordent Reach, Oral-B, and Fluorgard). Roughness was evaluated at time intervals: T 0 after 24 h in artificial saliva (baseline); T 60 after being in assigned immersion media for 1 min daily over 60 days; and T final after artificial aging (20,000 thermal cycles, 1,200,000 mechanical loading cycles, and continuous immersion for 1,825 min). Surface morphology was qualitatively analyzed by scanning electron microscopy (SEM) at T 60 and T final. Roughness data were submitted to analysis of variance for mixed repeated measures, Sidak, and Tukey tests at α=0.05. Micro-filled resin showed the highest roughness values. Fluoride solutions had no influence on roughness. Higher roughness values were observed after artificial aging. In SEM observations after the artificial aging, the specimens showed surface degradation, irrespective of immersion medium or type of composite. Nano-filled resin showed higher loss of resin matrix and protrusion of filler particles. Roughness was not influenced by fluoride solutions; however, it is material dependent and increases over time.

Influence of light curing units and fluoride mouthrinse on morphological surface and color stability of a nanofilled composite resin.

Composite resin is a dental material susceptible to color change over time which limits the longevity of restorations made with this material. The influence of light curing units and different fluoride mouthrinses on superficial morphology and color stability of a nanofilled composite resin was evaluated. Specimens (N = 150) were prepared and polished. The experimental groups were divided according to the type of light source (halogen and LED) and immersion media (artificial saliva, 0.05% sodium fluoride solution-manipulated, Fluordent Reach, Oral B, Fluorgard). Specimens remained in artificial saliva for 24-h baseline. For 60 days, they were immersed in solutions for 1 min. Color readout was taken at baseline and after 60 days of immersion. Surface morphology was analyzed by Scanning Electron Microscopy (SEM) after 60 days of immersion. Color change data were submitted to two-way Analysis of Variance and Tukey tests (α = 0.05). Surface morphology was qualitatively analyzed. The factor light source presented no significant variability (P = 0.281), the immersion media, significant variability (P < 0.001) and interaction between factors, no significant variability (P = 0.050). According to SEM observations, no difference was noted in the surface of the specimens polymerized by different light sources, irrespective of the immersion medium. It was concluded that the light source did not influence the color stability of composite, irrespective of the immersion media, and among the fluoride solutions analyzed, Fluorgard was the one that promoted the greatest color change, however, this was not clinically perceptible. The immersion media did not influence the morphology of the studied resin.

Effects of immersion media and repolishing on color stability and superficial morphology of nanofilled composite resin.

This study evaluated the influence of fluoride mouth rinses and repolishing on the superficial morphology and color stability of nanofilled resin. About 150 specimens were prepared and polished using aluminum oxide discs for 15 s with a pressure of 2 kg. The experimental groups were divided according to the immersion medium (artificial saliva, 0.5% sodium fluoride, Fluordent Reach, Oral B, Fluorgard) and repolishing procedure (without and with). The specimens were continuously immersed for 1 week. Thereafter, half of each sample was repolished. A color reading was performed after 24 h of immersion in the artificial saliva baseline, after continuous immersion, and after repolishing. The superficial morphology was examined using scanning electron microscopy (SEM) in a qualitative way. Color change (∆E) data were submitted to a mixed analysis of variance using a Shapiro-Wilk test (p>0.05 for the different immersion media) and Sidak's test (p<0.05 for the differences between groups). In the interaction between the repolishing and the immersion media, Fluorgard showed a statistical difference between the ∆E values with and without repolishing (p<0.0001). On the SEM observations, both Fluordent Reach and Fluorgard caused degradation of the superficial resinous matrix of the composite after continuous immersion. This matrix was removed after repolishing.

Chemical and morphological features of nanofilled composite resin: influence of finishing and polishing procedures and fluoride solutions.

This study evaluated the influence of finishing and polishing procedures and different fluoride solutions on superficial morphology and chemistry of the nanofilled composite resin Supreme XT (3M) through the EDX analysis and SEM evaluation. Circular specimens (n = 30) of 10 mm diameter and 2 mm thickness were prepared, with half of the sample assays finished and polished with Super-Snap® sandpaper. The experimental groups were divided according to the presence or absence of finishing and polishing and solutions (artificial saliva, 0.05% of manipulated sodium fluoride solution, Fluordent Reach, Oral B, Fluorgard). Specimens were immersed in each respective solution for 1 min per day, during 60 days and stored in artificial saliva at 37 ± 1°C between immersion periods. Topography and chemical analysis was qualitative. It was observed that specimens submitted to finishing and polishing procedures had lower superficial degradation. Fluoride solutions promoted superficial alterations on specimens, being the highest degradation obtained with Fluordent Reach. It can be concluded that finishing and polishing procedures and the immersion media influence the superficial morphology of composite resin tested; the Fluordent Reach was the fluoride solution that most affected the material's surface.

Chemical and morphological features of dental composite resin: influence of light curing units and immersion media.

AIMS: The study evaluated the influence of light curing units and immersion media on superficial morphology and chemistry of the nanofilled composite resin Supreme XT (3M) through the EDX analysis and SEM evaluation. Light curing units with different power densities and mode of application used were XL 3000 (480 mW/cm(2)), Jet Lite 4000 Plus (1230mW/cm(2)), and Ultralume Led 5 (790 mW/cm(2)) and immersion media were artificial saliva, Coke((R)), tea and coffee, totaling 12 experimental groups. Specimens (10 mm x 2 mm) were immersed in each respective solution for 5 min, three times a day, during 60 days and stored in artificial saliva at 37 degrees C +/- 1 degrees C between immersion periods. Topography and chemical analysis was qualitative. FINDINGS: Groups immersed in artificial saliva, showed homogeneous degradation of matrix and deposition of calcium at the material surface. Regarding coffee, there was a reasonable chemical degradation with loss of load particles and deposition of ions. For tea, superficial degradation occurred in specific areas with deposition of calcium, carbon, potassium and phosphorus. For Coke((R)), excessive matrix degradation and loss of load particles with deposition of calcium, sodium, and potassium. CONCLUSION: Light curing units did not influence the superficial morphology of composite resin tested, but the immersion beverages did. Coke((R)) affected material's surface more than did the other tested drinks.