In vitro study of structural and chemical changes in two reciprocating files after multiple reuses and sterilisation.

To use scanning electron microscopy and energy dispersive X-ray spectroscopy to evaluate the metallurgical-chemical changes of WaveOne Gold (WOG) and R-Motion (RM), after multiple uses. The instruments were divided into groups (n = 8): WOG and RM-control groups, new instruments; WOG1 and RM1; WOG2 and RM2; WOG3 and RM3 after instrumentation of 1, 2 or 3 molars, respectively. Burrs occurred mainly in the control group and after the first use. The RM files were found to have a higher nickel content, which increased during reuse, and a decrease in oxygen content with increasing reuse, in addition to calcium impregnation, which occurred in greater amounts in the corrosion areas in the WOG files. The presence of topographic and chemical changes was demonstrated, indicating that caution should be taken when reusing endodontic instruments to avoid fractures.

The physical and evolutionary energy landscapes of devolved protein sequences corresponding to pseudogenes.

Protein evolution is guided by structural, functional, and dynamical constraints ensuring organismal viability. Pseudogenes are genomic sequences identified in many eukaryotes that lack translational activity due to sequence degradation and thus over time have undergone "devolution." Previously pseudogenized genes sometimes regain their protein-coding function, suggesting they may still encode robust folding energy landscapes despite multiple mutations. We study both the physical folding landscapes of protein sequences corresponding to human pseudogenes using the Associative Memory, Water Mediated, Structure and Energy Model, and the evolutionary energy landscapes obtained using direct coupling analysis (DCA) on their parent protein families. We found that generally mutations that have occurred in pseudogene sequences have disrupted their native global network of stabilizing residue interactions, making it harder for them to fold if they were translated. In some cases, however, energetic frustration has apparently decreased when the functional constraints were removed. We analyzed this unexpected situation for Cyclophilin A, Profilin-1, and Small Ubiquitin-like Modifier 2 Protein. Our analysis reveals that when such mutations in the pseudogene ultimately stabilize folding, at the same time, they likely alter the pseudogenes' former biological activity, as estimated by DCA. We localize most of these stabilizing mutations generally to normally frustrated regions required for binding to other partners.

Optimizing cryopreservation conditions for use of fucosylated human mesenchymal stromal cells in anti-inflammatory/immunomodulatory therapeutics.

Mesenchymal stem/stromal cells (MSCs) are being increasingly used in cell-based therapies due to their broad anti-inflammatory and immunomodulatory properties. Intravascularly-administered MSCs do not efficiently migrate to sites of inflammation/immunopathology, but this shortfall has been overcome by cell surface enzymatic fucosylation to engender expression of the potent E-selectin ligand HCELL. In applications of cell-based therapies, cryopreservation enables stability in both storage and transport of the produced cells from the manufacturing facility to the point of care. However, it has been reported that cryopreservation and thawing dampens their immunomodulatory/anti-inflammatory activity even after a reactivation/reconditioning step. To address this issue, we employed a variety of methods to cryopreserve and thaw fucosylated human MSCs derived from either bone marrow or adipose tissue sources. We then evaluated their immunosuppressive properties, cell viability, morphology, proliferation kinetics, immunophenotype, senescence, and osteogenic and adipogenic differentiation. Our studies provide new insights into the immunobiology of cryopreserved and thawed MSCs and offer a readily applicable approach to optimize the use of fucosylated human allogeneic MSCs as immunomodulatory/anti-inflammatory therapeutics.

Non-invasive meat quality assessment: Exploring the potential of ocular infrared thermography to predict ultimate pH in Nellore beef cattle.

This study investigated the use of infrared thermography (IRT) to identify the dark, firm, and dry (DFD) phenomenon in Brazilian beef, which is a significant concern for the industry because of its inferior quality and reduced shelf life. This study examined 113 Nellore bulls and analyzed their minimum and maximum ocular temperatures using IRT. The results highlight the efficacy of thermal images (IRTmax) as a significant predictor, with R2 values ranging from 0.84 to 0.88 for calibration models. The inclusion of parameters such as glucose and lactate further enhanced prediction accuracy. The models also revealed that the combination of features, such as lightness (L*), redness (a*), and yellowness (b*), contributed to the precise prediction of pHu, with an R2 of 0.88. In model validation, RMSEP ranged from 0.104 to 0.158, indicating good generalization capability. The RPD, ranging from 1.7 to 2.6, suggests satisfactory quantitative prediction. The statistical significance of all models, evidenced by P-values <0.001, strengthens the reliability of the results. In conclusion, the models support the use of IRT as a tool for identifying pHu alterations in carcasses. When combined with blood parameters, they may exhibit even greater efficiency in predicting pHu in Nelore cattle carcasses, highlighting the potential applicability of these methods in the beef industry.

Aesculus hippocastanum extract and the main bioactive constituent ß-escin as antivirals agents against coronaviruses, including SARS-CoV-2.

Respiratory viruses can cause life-threatening illnesses. The focus of treatment is on supportive therapies and direct antivirals. However, antivirals may cause resistance by exerting selective pressure. Modulating the host response has emerged as a viable therapeutic approach for treating respiratory infections. Additionally, considering the probable future respiratory virus outbreaks emphasizes the need for broad-spectrum therapies to be prepared for the next pandemics. One of the principal bioactive constituents found in the seed extract of Aesculus hippocastanum L. (AH) is ß-escin. The clinical therapeutic role of ß-escin and AH has been associated with their anti-inflammatory effects. Regarding their mechanism of action, we and others have shown that ß-escin and AH affect NF-κB signaling. Furthermore, we have reported the virucidal and broad-spectrum antiviral properties of ß-escin and AH against enveloped viruses such as RSV, in vitro and in vivo. In this study, we demonstrate that ß-escin and AH have antiviral and virucidal activities against SARS-CoV-2 and CCoV, revealing broad-spectrum antiviral activity against coronaviruses. Likewise, they exhibited NF-κB and cytokine modulating activities in epithelial and macrophage cell lines infected with coronaviruses in vitro. Hence, ß-escin and AH are promising broad-spectrum antiviral, immunomodulatory, and virucidal drugs against coronaviruses and respiratory viruses, including SARS-CoV-2.

The nuclei of human adult stem cells can move within the cell and generate cellular protrusions to contact other cells.

BACKGROUND: The neuronal transdifferentiation of adult bone marrow cells (BMCs) is still considered an artifact based on an alternative explanation of experimental results supporting this phenomenon obtained over decades. However, recent studies have shown that following neural induction, BMCs enter an intermediate cellular state before adopting neural-like morphologies by active neurite extension and that binucleated BMCs can be formed independent of any cell fusion events. These findings provide evidence to reject the idea that BMC neural transdifferentiation is merely an experimental artifact. Therefore, understanding the intermediate states that cells pass through during transdifferentiation is crucial given their potential application in regenerative medicine and disease modelling. METHODS: In this study, we examined the functional significance of the variety of morphologies and positioning that cell nuclei of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) can adopt during neural-like differentiation using live-cell nuclear fluorescence labelling, time-lapse microscopy, and confocal microscopy analysis. RESULTS: Here, we showed that after neural induction, hBM-MSCs enter an intermediate cellular state in which the nuclei are able to move within the cells, switching shapes and positioning and even generating cellular protrusions as they attempt to contact the cells around them. These findings suggest that changes in nuclear positioning occur because human cell nuclei somehow sense their environment. In addition, we showed the process of direct interactions between cell nuclei, which opens the possibility of a new level of intercellular interaction. CONCLUSIONS: The present study advances the understanding of the intermediate stage through which hBM-MSCs pass during neural transdifferentiation, which may be crucial to understanding the mechanisms of these cell conversion processes and eventually harness them for use in regenerative medicine. Importantly, our study provides for the first time evidence that the nuclei of hBM-MSC-derived intermediate cells somehow sense their environment, generating cellular protrusions to contact other cells. In summary, human mesenchymal stromal cells could not only help to increase our understanding of the mechanisms underlying cellular plasticity but also facilitate the exact significance of nuclear positioning in cellular function and in tissue physiology.

Bone Resorption in Apical Periodontitis Enhanced by Cigarette Smoke Inhalation: Histometric, Immunohistochemical, and Microtomographic Analysis in Rats.

INTRODUCTION: This study evaluated the effects of cigarette smoke inhalation (CSI) on apical periodontitis (AP) induced in rats by histometric, immunohistochemical, and microtomographic analysis. METHODS: A total of 32 male Wistar rats were divided into 4 experimental groups (n = 8): control, CSI, AP, and CSI + AP. Rats in the CSI and CSI + AP groups inhaled cigarette smoke by remaining inside a smoking chamber for 8 minutes 3 times a day for 50 days. After 20 days of smoke inhalation, rats in the AP and CSI + AP groups had the pulp of their first right lower molar exposed to induce AP. Blood was collected on day 50 to evaluate nicotine and serum cotinine levels. The animals' mandibles were removed for histologic processing to evaluate bone resorption by histometric, immunohistochemical (receptor activator of nuclear factor kappa B ligand/osteoprotegerin), and microtomographic analysis. The Student t test was applied. RESULTS: Histometric analysis showed a larger area of bone resorption (P < .05) and microtomographic analysis found greater resorption volume (P < .001) for the CSI + AP group compared with the AP group. The CSI + AP group presented a high RANKL immunostaining pattern compared with the AP group (P < .001). CONCLUSIONS: CSI increased bone resorption caused by AP.

Evaluation of the Ability of 3 Reciprocating Instruments to Remove Obturation Material: A Micro-Computed Tomography Study.

INTRODUCTION: The purpose of this study was to evaluate the ability of the Reciproc, Reciproc Blue, and WaveOne Gold systems to remove filling material during endodontic retreatment of extracted human mandibular premolars. METHODS: Thirty-nine teeth were instrumented with the Protaper Universal System to the F3 file and filled with the Tagger hybrid technique using an F3 gutta-percha cone and AH Plus cement. At the end of this period, the teeth were scanned with micro-computed tomography before and after removal of the filling material from the root canals. The teeth were divided into 3 groups (n = 13) based on the apical volume, depending on the systems used to remove the filling material. Group GR: Reciproc 40/.06; Group GRB: Reciproc Blue 40/.06; and Group GWG: WaveOne Gold 35/.06. The results were statistically analyzed using the tests of Kruskal-Wallis, Duncan, and analysis of variance at a significance level of 5%. RESULTS: The results showed that there were no significant differences between the amounts of filling material removed, either for the apical and middle regions alone or in the overall evaluation for the 3 groups (P = .97). The time evaluation statistically showed that the GR and GWG groups required less time to clean the root canals than the GRB group. CONCLUSIONS: Reciproc R40 files and WaveOne Gold Medium files required less time for endodontic treatment than Reciproc Blue R40 files. There was no difference in the ability to remove obturation material between the 3 instruments. No instrument was able to completely remove the filling material from the root canals.

Comparative in vitro analysis of the antifungal activity of different calcium silicate-based endodontic sealers

Aim: This study aimed to perform an in vitro comparative analysis of the antifungal activity of different calcium silicate-based endodontic sealers against three fungal species. Methods: The antifungal properties of three calcium silicate-based sealers were tested: Bio-C Sealer, Cambiar a Sealer Plus BC, and MTA-Fillapex. Two commonly used sealers were used as controls: AH Plus and Endomethasone. An agar diffusion test was performed to analyze the antifungal activity of the sealers against Candida albicans, Candida glabrata, Candida tropicalis, and a mixed microbial culture medium. The results were analyzed using ANOVA (p <0.05). Results: Endomethasone exhibited the highest inhibition against all strains examined, maintaining a consistent level of inhibition throughout 7 days. MTA-Fillapex demonstrated the best performance among the calcium silicate-based sealers for the three fungal species (p < 0.05), maintaining stable values over the 7 days, surpassing that of Endomethasone. Nevertheless, MTA-Fillapex only exhibited antimicrobial effect against the mixed culture for the first 24 hours, and no antimicrobial activity was observed at 48 hours, being surpassed by all tested sealers (p < 0.05). Conclusion: Of all silicate-based sealers tested, only MTA-Fillapex exhibited promising antifungal activity. Nevertheless, care must be taken when extrapolating these results, as MTA-Fillapex exhibited poor antimicrobial activity when tested in mixed microbial cultures

Effect of obturation technique on dentinal tubule penetration of two tricalcium silicate-based sealers: Ex vivo study.

The aim of this study was to assess by confocal laser microscope the depth of dentinal tubule penetration of two tricalcium silicate-based sealers promoted by two obturation techniques in curved canals compared with AHPlus. One hundred and twenty canals were divided into six groups (n = 20): BCSC-Bio-C Sealer (BC) and single-cone technique (SC); BCCW-BC and continuous condensation wave (CW); TFSC-Total Fill (TF) and SC; TFCW-TF and CW; AHSC-AH Plus (AH) and SC; AHCW-AH and CW. Data were analysed using the three-way ANOVA and Tukey's test (α = 5%). Penetration depth was significantly greater for TFCW than TFSC and greater for AHCW than AHSC (p < 0.05). There was no significant difference between BCCW and BCSC (p > 0.05). The penetration of TF was significantly greater (p < 0.05). The CW technique promoted greater intratubular penetration, except for the BC sealer.

Evaluation of the apical sealing of an eggshell hydroxyapatite-based sealer.

Background: The success of endodontic treatment can be influenced by the type of endodontic sealer used, as certain sealers may be prone to apical microleakage, leading to treatment failure. The limitations of currently available sealers necessitate the development of new materials to improve the success rate of endodontic treatment. Therefore, the objective of this study was to assess the apical microleakage of newly developed hydroxyapatite-based endodontic sealers, including one derived from eggshells, and compare them with other commercially available sealers. Material and Methods: Eighty-five extracted human upper anterior teeth were selected for this study. The teeth were divided into 5 experimental groups and 2 control groups. The experimental groups were designated as follows: (1) HPSINT - obturated with gutta-percha cone and synthetic hydroxyapatite-based sealer, (2) BIOC - obturated with gutta-percha cone and Bio C-Sealer sealer, (3) AHPLUS-BC - obturated with gutta-percha cone and AHPLUS Bioceramic sealer, (4) AHP - obturated with gutta-percha cone and AHPLUS sealer, and (5) HPO - obturated with gutta-percha cone and sealer based on hydroxyapatite extracted from eggshells. Additionally, there were positive and negative control groups consisting of instrumented teeth filled with gutta-percha cones without any sealer and instrumented teeth without any filling, respectively. Methylene blue dye penetration was used to assess apical microleakage. Descriptive statistical analysis and Shapiro-Wilk normality test were applied to the observed results. As the samples followed a normal distribution, the ANOVA test was applied. Results: The control groups confirmed the validity of the experimental method, while the experimental groups showed varying degrees of dye penetration. The group obturated with Bio C-Sealer exhibited the highest mean apical microleakage, while AHPLUS Bioceramic sealer demonstrated lower mean than AHPLUS sealer and sealer based on hydroxyapatite extracted from eggshells (p<0.05). Finally, there was no difference between the synthetic hydroxyapatite-based sealer and AHPLUS Bioceramic sealer, AHPLUS sealer and sealer based on hydroxyapatite extracted from eggshells (p>0.05). No significant difference was observed between the hydroxyapatite-based sealers and the AHPLUS-BC sealer. Conclusions: The results of this study suggest that the newly developed hydroxyapatite-based endodontic sealers, including the one derived from eggshells, may have a lower risk of apical microleakage compared to other commercially available sealers. These findings highlight the potential of hydroxyapatite-based sealers to improve the success rate of endodontic treatment. Further research and clinical studies are warranted to validate these results and explore the long-term effects of these novel sealers. Key words:Endodontic treatment, apical microleakage, endodontic sealer, hydroxyapatite, eggshell-derived sealer.

Interdisciplinary Therapy Improves the Mediators of Inflammation and Cardiovascular Risk in Adolescents with Obesity.

Obesity is associated with inflammation and an increased risk of cardiovascular disease and premature mortality, as well as a range of other conditions. Obesity is a growing global problem, not only in adults, but also in children and adolescents. Therefore, the present study aimed to assess the effects of a one-year interdisciplinary intervention on the cardiometabolic and inflammatory profiles of adolescents with obesity. Twenty-two adolescents completed the intervention, which included clinical, nutritional, psychological and physical exercise counselling. Body composition, and metabolic, inflammatory, and cardiovascular risk biomarkers were analyzed before and after one year of intervention. Visceral and subcutaneous fat were determined ultrasonographically. The homeostasis model assessment of insulin resistance (HOMA-IR) and the quantitative insulin sensitivity check index (QUICKI) equation were used to estimate insulin resistance and insulin sensitivity, respectively. A reduction in body mass, adiposity, glucose, and insulin and an improved lipid profile were observed after the therapy. Hyperleptinemia was reduced from 77.3% to 36.4%. Plasminogen activator inhibitor-1 (PAI-1), intercellular adhesion molecule 1 (ICAM-1), leptin, the leptin/adiponectin ratio, and the adiponectin/leptin ratio were also significantly improved. Metabolic changes were associated with a reduction in visceral fat and waist circumference, and adiponectin and the leptin/adiponectin ratio were associated with HOMA-IR. The interdisciplinary therapy promoted improvements in hyperleptinemia and metabolic, inflammatory, and cardiovascular biomarkers.

Assessing the cyclic fatigue resistance and sterilization effects on replica-like endodontic instruments compared to Reciproc Blue.

This study aimed to evaluate the effect of the number of uses and autoclave sterilization on the cyclic fatigue resistance of four replica-like instruments RC Blue; Only One File Blue; Recip One Blue; and Micro Blue compared to the original system Reciproc Blue (VDW, Munich, Germany). The instruments were analyzed by scanning electron microscope (SEM) before being used in root canal instrumentation (baseline). Fifty molars were instrumented by ten instruments (n=10). After sterilization in an autoclave, the  instruments were analyzed by SEM. This procedure was repeated twice more using different molars, totaling 3 rounds of instrumentation, sterilization and SEM analysis. Then, ten different instruments from each brand were tested for cyclic fatigue resistance. Number of uses data were analyzed using Chi-squared analysis, and cyclic fatigue data were analyzed by one-way Anova followed by Tukey's test as the data had normal distribution. The fracture times for all systems had no significant difference, but Micro Blue had significantly lower values than the other systems (p < 0.05). The SEM analysis showed distortions in the instruments after the 3rd use. Therefore, all tested instruments except of Micro Blue have similar resistance to cyclic fatigue, and all are reliable for use in up to 2-cases.

Effects of cigarette smoke inhalation on the immune-inflammatory profile of experimental apical periodontitis in rats.

OBJECTIVE: To evaluate the effects of cigarette smoke inhalation on the immune-inflammatory profile of experimental apical periodontitis in rats. METHODOLOGY: In total, 32 male Wistar rats were divided into four groups (n = 8): AP-induced apical periodontitis; S-cigarette smoke inhalation; APS-induced AP and cigarette smoke inhalation; and C (control)-neither AP nor cigarette smoke inhalation. To induce cigarette smoke inhalation, the animals were kept in a chamber filled with tobacco smoke for 8 min thrice a day for 50 days. AP was induced 20 days after inhalation initiation by exposing their coronary pulp to their oral environment for 30 days. After animals were euthanized, their right hemimaxillae were removed for histopathological, semi-quantitative and immunohistochemical (F4/80, CD206 and iNOS) analyses. RESULTS: Quantitative data showed a moderate number of inflammatory infiltrates in AP and an intense number in APS (p < .05). Comparing F4/80+ cells showed no statistically significant differences among groups, but we found more CD206+ cells in AP than in C and S (p > .05). INOS+ immunostaining showed a significant increase in AP and APS, when compared with C and S (p < .05). APS had more iNOS+ cells than AP (p < .05). CONCLUSION: Cigarette smoke inhalation worsened AP, leading to a predominantly pro- inflammatory profile in our experimental model.

Biocompatibility, Biomineralization and Induction of Collagen Maturation with the Use of Calcium Hydroxide and Iodoform Intracanal Dressing.

Biocompatibility and biomineralization of root canal dressings are important requirements for periapical healing. This study evaluated the inflammatory response, biomineralization and tissue repair by collagen fiber maturation in the subcutaneous tissue of rats. Eighteen Wistar rats (n = 6) received subcutaneous implants: calcium hydroxide + propylene glycol [CH+P], calcium hydroxide + propylene glycol + iodoform [CH+P+I], iodoform + carbowax [I+Cwax] and carbowax [Cwax]. Extra empty tubes were used as a control [C]. After 7, 15 and 30 days, the implants were removed with surrounding tissue for staining of hematoxylin-eosin, Von Kossa, picrosirius red and without staining for analysis under polarized light. Results were analyzed via Kruskal-Wallis followed by Dunn testing for nonparametric data and ANOVA followed by a Tukey post hoc test for parametric data (p < 5%). At 7 days, all groups showed a moderate inflammatory reaction and thick fibrous capsule, except the [Cwax] group, with a severe inflammatory infiltrate (p < 0.05). After 15 days, all groups but control had a decrease in inflammatory response. At 30 days, all groups presented a mild reaction and thin fibrous capsule (p > 0.05). Only groups containing calcium hydroxide were found to be positive using Von Kossa staining and polarized light in all periods. At 7 days, all groups showed a higher proportion of immature fibers. At 15 days, the [CH+P] and [Cwax] groups increased their proportion of mature/immature fibers. At 30 days, only the [CH+P] group presented a significant prevalence of mature collagen fibers (p < 0.05). All groups showed biocompatibility, but only groups containing calcium hydroxide induced biomineralization. The addition of iodoform delayed tissue healing.

Two synthetic steroid analogs reduce human respiratory syncytial virus replication and the immune response to infection both in vitro and in vivo.

HRSV is responsible for many acute lower airway infections and hospitalizations in infants, the elderly and those with weakened immune systems around the world. The strong inflammatory response that mediates viral clearance contributes to pathogenesis, and is positively correlated with disease severity. There is no specific effective therapy on hand. Antiviral synthetic stigmastanes (22S, 23S)-22,23-dihydroxystigmast-4-en-3-one (Compound 1) and 22,23-dihydroxystigmasta-1,4-dien-3-one (Compound 2) have shown to be active inhibiting unrelated virus like Herpes Simplex type 1 virus (HSV-1) and Adenovirus, without cytotoxicity. We have also shown that Compound 1 modulates the activation of cell signaling pathways and cytokine secretion in infected epithelial cells as well as in inflammatory cells activated by nonviral stimuli. In the present work, we investigated the inhibitory effect of both compounds on HRSV replication and their modulatory effect on infected epithelial and inflammatory cells. We show that compounds 1 and 2 inhibit in vitro HRSV replication and propagation and reduce cytokine secretion triggered by HRSV infection in epithelial and inflammatory cells. The compounds reduce viral loads and inflammatory infiltration in the lungs of mice infected with HRSV.

Broad-Spectrum Antiviral Effect of Cannabidiol Against Enveloped and Nonenveloped Viruses.

Introduction: Cannabidiol (CBD), the main non-psychoactive cannabinoid of the Cannabis sativa plant, is a powerful antioxidant compound that in recent years has increased interest due to causes effects in a wide range of biological functions. Zika virus (ZIKV) is a virus transmitted mainly by the Aedes aegypti mosquitoes, which causes neurological diseases, such as microcephaly and Guillain-Barre syndrome. Although the frequency of viral outbreaks has increased recently, no vaccinations or particular chemotherapeutic treatments are available for ZIKV infection. Objectives: The major aim of this study was to explore the in vitro antiviral activity of CBD against ZIKV, expanding also to other dissimilar viruses. Materials and Methods: Cell cultures were infected with enveloped and nonenveloped viruses and treated with non-cytotoxic concentrations of CBD and then, viral titers were determined. Additionally, the mechanism of action of the compound during ZIKV in vitro infections was studied. To study the possible immunomodulatory role of CBD, infected and uninfected Huh-7 cells were exposed to 10 µM CBD during 48 h and levels of interleukins 6 and 8 and interferon-beta (IFN-ß) expression levels were measured. On the other hand, the effect of CBD on cellular membranes was studied. For this, an immunofluorescence assay was performed, in which cell membranes were labeled with wheat germ agglutinin. Finally, intracellular cholesterol levels were measured. Results: CBD exhibited a potent antiviral activity against all the tested viruses in different cell lines with half maximal effective concentration values (CE50) ranging from 0.87 to 8.55 µM. Regarding the immunomodulatory effect of CBD during ZIKV in vitro infections, CBD-treated cells exhibited significantly IFN-ß increased levels, meanwhile, interleukins 6 and 8 were not induced. Furthermore, it was determined that CBD affects cellular membranes due to the higher fluorescence intensity that was observed in CBD-treated cells and lowers intracellular cholesterol levels, thus affecting the multiplication of ZIKV and other viruses. Conclusions: It was demonstrated that CBD inhibits structurally dissimilar viruses, suggesting that this phytochemical has broad-spectrum antiviral effect, representing a valuable alternative in emergency situations during viral outbreaks, like the one caused by severe acute respiratory syndrome coronavirus 2 in 2020.

Biological investigation of resinous endodontic sealers containing calcium hydroxide.

The purpose of this study was to evaluate, in vivo, the biocompatibility, biomineralization, collagen maturation and the in vitro antibacterial and cytotoxicity of resinous endodontic sealers containing calcium hydroxide. Forty rats were implanted with polyethylene tubes containing Sealer 26, Sealer Plus, Dia-ProSeal and an empty tube, examined after 7, 15, 30 and 60 days. Antimicrobial activity was evaluated against Enterococcus faecalis by Agar Diffusion Test (ADT) through inhibition zones. For cytotoxicity, undifferentiated pulp cells (OD-21) were cultured and assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, exposed to dilution of serial extracts at 6, 24, 48h. Cytotoxicity was analyzed by two-way ANOVA and Bonferroni correction. Kruskal-Wallis test followed by Dunn test was performed for nonparametric data (p<0.05). MTT assay revealed cell proliferation affected by sealers extract in all periods (p<0.0001), except for Dia-Proseal and Sealer Plus ⅛ dilution. Subcutaneous analysis showed at day 7th moderate inflammatory infiltration. After 30 days, Sealer 26 still showed moderate inflammatory infiltrate compared to mild inflammation from control and Dia-ProSeal (p = 0.006). At day 60th, all groups showed similar mild inflammatory infiltrate (p>0.05). Sealer 26 induced more biomineralization than other sealers in all periods. At 7 and 15 days, all sealers had significant percentage of immature collagen fibers. After 60 days Sealer 26 showed more mature fibers compared to other sealers (p<0.001). All sealers had a smaller zone of inhibition than chlorhexidine, but with no significant difference among any group (p>0.05). All sealers showed satisfactory biological responses with in vitro/in vivo biocompatibility and antimicrobial activity against planktonic bacteria. Sealer 26 induced more biomineralization than Sealer Plus and Dia-ProSeal.

Analysis of antimicrobial efficacy of sodium hypochlorite and ozonated water against biofilm in oval canals.

This in vitro study compared the antimicrobial efficacy of 2.5% sodium hypochlorite (NaOCl) and 8 µg/mL ozonated water agitated by passive ultrasonic irrigation (PUI) or PUI combined with EndoActivator (EA) against mature multispecies biofilm. One hundred and five oval-shaped mandibular premolars were instrumented, sterilized, and inoculated with Enterococcus faecalis, Candida albicans, and Staphylococcus aureus, divided into: control group - saline; O3 group - ozonated water; O3 PUI group - ozonated water with PUI agitation; O3 PUI+EA group - ozonated water with PUI+EA agitation; NaOCl group - NaOCl; NaOCl PUI group - NaOCl with PUI agitation; and NaOCl PUI+EA group - NaOCl with PUI+EA agitation. Microbiological samples were collected before (S1) and after (S2) the disinfection procedures and the data were statistically analyzed using the Kruskal-Wallis test. In the culture method, there was significant disinfection in the O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). The combination of NaOCl with PUI+EA reduced microbial counts to zero (p˂0.05). In the qPCR method, there was a significant reduction in the total count of viable microorganisms in the O3 PUI, O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). It can be concluded that 2.5% NaOCl with and without agitation, as well as 8 µg/mL ozonated water with its action enhanced by the agitation techniques, were effective in root canal disinfection, and their antimicrobial efficacy is related to the microorganisms present in the biofilm.

Eight Weeks of Physical Training Decreases 2 Years of DNA Methylation Age of Sedentary Women.

Purpose: The acceleration of epigenetic age is a predictor of mortality and contributes to the increase in chronic diseases. Adherence to a healthy lifestyle is a strategy to reduce epigenetic age. The present study aimed to determine whether eight weeks of combined (aerobic and strength) training (CT) can influence the epigenetic age of women between 50 and 70 years old and the differences in sites and methylated regions. Methods: Eighteen women (AARLow: lower age acceleration residual, n = 10; AARHigh: higher age acceleration residual, n = 8) participated in a combined exercise training program (60 minutes, 3× a week) for eight weeks. DNA was extracted from whole blood using the salting out technique. DNA methylation was performed using the array technique (Illumina's Infinium Methylation BeadChip 850k). We used the DNA Methylation Age Calculator platform to calculate the biological epigenetic age. Two-way ANOVA followed by FISHER LSD posthoc was Applied, adopting p < .05. Results: After eight weeks of CT, there were no changes to the epigenetic age acceleration for the AARLow group (PRE: -2.3 ± 3.2 to POST: -2.3 ± 3.6). However, the AARHigh group significantly decreased the age acceleration (PRE: 3.6 ± 2.6 to POST: 2.2 ± 2.7) (group effect, p = .01; time effect, p = .31; group vs. time effect, p = .005). Conclusion: CT for eight weeks benefits the epigenetic clock of women with the most accelerated age.