Effect of Various Surface Treatments of Zirconia on its Adhesive Properties to Dentin: An In Vitro Study.

AIM: To assess the effectiveness of various surface treatments and adhesives on the bond strength of zirconia-based ceramic to dentin. MATERIALS AND METHODS: Eighty samples of zirconia were subjected to the four-surface treatment protocols (sandblasting, 48% hydrofluoric acid (HF), 48% hydrofluoric acid + 70% nitric acid (HNO3) and no treatment (control) following which the samples from each group were subdivided into two subgroups (n = 10) based on the resin cement employed for cementation (RelyX U200 and G-Cem Linkforce). The bonded specimens were subjected to shear stress to measure the bond strength using Universal testing machine. To test the difference in bond strength among the eight study groups, the Kruskal-Wallis ANOVA test was applied and for comparison between cements in each group, Mann-Whitney U test was applied. RESULTS: The highest bond strength values were observed for 48% HF group cemented with G-Cem Linkforce resin cement (16.220 ± 1.574) and lowest for control group-RelyX (4.954 ± 0.972). G-Cem cement showed higher bond strength than RelyX for all surface treatments except 48% HF + 70% nitric acid. CONCLUSION: It can be inferred that 48% HF can etch zirconia and generate a porous structure that proves to be beneficial for bonding. CLINICAL SIGNIFICANCE: The increasing demand for esthetics has led to the replacement of metal-ceramic materials with zirconia-based ceramics. However, the chemical inertness of zirconia to various conventional surface treating agents has continuously challenged researchers to discover a new surface treatment protocol that could enhance the bond strength of zirconia. How to cite this article: Yenamandra MS, Joseph A, Singh P, et al. Effect of Various Surface Treatments of Zirconia on its Adhesive Properties to Dentin: An In Vitro Study. J Contemp Dent Pract 2024;25(3):226-230.

Chromosome organization shapes replisome dynamics in Caulobacter crescentus.

DNA replication in bacteria takes place on highly compacted chromosomes, where segregation, transcription, and repair must occur simultaneously. Within this dynamic environment, colocalization of sister replisomes has been observed in many bacterial species, driving the hypothesis that a physical linker may tether them together. However, replisome splitting has also been reported in many of the same species, leaving the principles behind replisome organization a long-standing puzzle. Here, by tracking the replisome ß-clamp subunit in live Caulobacter crescentus, we find that rapid DNA segregation can give rise to a second focus which resembles a replisome, but does not replicate DNA. Sister replisomes can remain colocalized, or split apart to travel along DNA separately upon disruption of chromosome inter-arm alignment. Furthermore, chromosome arm-specific replication-transcription conflicts differentially modify replication speed on the two arms, facilitate the decoupling of the two replisomes. With these observations, we conclude that the dynamic chromosome organization flexibly shapes the organization of sister replisomes, and we outline principles which can help to reconcile previously conflicting models of replisome architecture.

Mechanistic insight into the repair of C8-linked pyrrolobenzodiazepine monomer-mediated DNA damage.

Pyrrolobenzodiazepines (PBDs) are naturally occurring DNA binding compounds that possess anti-tumor and anti-bacterial activity. Chemical modifications of PBDs can result in improved DNA binding, sequence specificity and enhanced efficacy. More recently, synthetic PBD monomers have shown promise as payloads for antibody drug conjugates and anti-bacterial agents. The precise mechanism of action of these PBD monomers and their role in causing DNA damage remains to be elucidated. Here we characterized the damage-inducing potential of two C8-linked PBD bi-aryl monomers in Caulobacter crescentus and investigated the strategies employed by cells to repair the same. We show that these compounds cause DNA damage and efficiently kill bacteria, in a manner comparable to the extensively used DNA cross-linking agent mitomycin-C (MMC). However, in stark contrast to MMC which employs a mutagenic lesion tolerance pathway, we implicate essential functions for error-free mechanisms in repairing PBD monomer-mediated damage. We find that survival is severely compromised in cells lacking nucleotide excision repair and to a lesser extent, in cells with impaired recombination-based repair. Loss of nucleotide excision repair leads to significant increase in double-strand breaks, underscoring the critical role of this pathway in mediating repair of PBD-induced DNA lesions. Together, our study provides comprehensive insights into how mono-alkylating DNA-targeting therapeutic compounds like PBD monomers challenge cell growth, and identifies the specific mechanisms employed by the cell to counter the same.

Comparative Evaluation of Root Canal Centering Ability of ProTaper, Mtwo, WaveOne, and Reciproc Using Cone-beam Computed Tomography: In Vitro Study.

AIM: To compare the centering ability of rotating (ProTaper and Mtwo) and reciprocating (WaveOne and Reciproc) file systems using cone-beam computed tomography (CBCT). MATERIALS AND METHODS: Eighty extracted human mandibular molars with curvature within 15-45° were selected and randomly divided into four groups (n = 20): group I (ProTaper), group II (Mtwo), group III (WaveOne), and group IV (Reciproc). The selected teeth were arranged in a template, and pre-instrumentation and post-instrumentation CBCT scans were taken using Kodak Carestream CS 9300 machine. The centering ability was measured in four planes namely, at furcation, 3 mm apical to furcation (coronal), 6 mm apical to furcation (middle), and 3 mm coronal to the apex (apical), wherein dentin thickness was measured from pre- and post-instrumentation CBCT scans and recorded for each canal (mesiobuccal and mesiolingual) separately and statistically analyzed. RESULTS: Mtwo and Reciproc remained better centered followed by WaveOne and least by ProTaper in different-level comparisons. In the mesiodistal dimension (MD) at the 3 mm, 6 mm, and 3 mm apical level, Mtwo and Reciproc showed better centering, meanwhile, in the buccolingual dimension, only in the 3 mm apical level, Mtwo and Reciproc remain better centered. A significant difference was seen between the group and type of canal. CONCLUSION: Mtwo and Reciproc showed better centered preparation than ProTaper and WaveOne. Significant differences were seen between the groups and types of canal. CLINICAL SIGNIFICANCE: Root canal instrumentation should maintain the original canal anatomy. The proper enlargement keeping in mind the remaining strength in the tooth structure is essentially decided by how the selected instrument is centered. The alloy used for instrument manufacture and its design (taper, cross-section, and tip) will influence the centered preparation. The type of instrument and the instrumentation techniques should be chosen based on root canal anatomy.

Coordination between nucleotide excision repair and specialized polymerase DnaE2 action enables DNA damage survival in non-replicating bacteria.

Translesion synthesis (TLS) is a highly conserved mutagenic DNA lesion tolerance pathway, which employs specialized, low-fidelity DNA polymerases to synthesize across lesions. Current models suggest that activity of these polymerases is predominantly associated with ongoing replication, functioning either at or behind the replication fork. Here we provide evidence for DNA damage-dependent function of a specialized polymerase, DnaE2, in replication-independent conditions. We develop an assay to follow lesion repair in non-replicating Caulobacter and observe that components of the replication machinery localize on DNA in response to damage. These localizations persist in the absence of DnaE2 or if catalytic activity of this polymerase is mutated. Single-stranded DNA gaps for SSB binding and low-fidelity polymerase-mediated synthesis are generated by nucleotide excision repair (NER), as replisome components fail to localize in the absence of NER. This mechanism of gap-filling facilitates cell cycle restoration when cells are released into replication-permissive conditions. Thus, such cross-talk (between activity of NER and specialized polymerases in subsequent gap-filling) helps preserve genome integrity and enhances survival in a replication-independent manner.

Evaluation of the Antimicrobial Efficacy of Elettaria cardamomum Oil, Trachyspermum ammi Oil and 5% Sodium Hypochlorite Against Enterococcus faecalis Biofilm Formed on Tooth Substrate.

CONTEXT: The usual cause of nonfulfillment of endodontic therapy is the persistence of microorganisms in the root canal system due to ineffective disinfection. Enterococcus faecalis is one of the most prevalently isolated microorganisms following a failure in root canal treatments. Sodium hypochlorite is among the most effectively used irrigant solutions but has many shortcomings. Herbal alternatives for sodium hypochlorite might prove to be superior due to their high antimicrobial activity, biocompatibility, and their antioxidant and anti-inflammatory properties. AIMS: This study is aimed to evaluate the antimicrobial efficacy of Trachyspermum ammi oil and Elettaria cardamomum oil against 2-week-old and 4-week-old E. faecalis biofilms formed on tooth substrate. SETTINGS AND DESIGN: A pure culture of E. faecalis was grown on brain heart infusion agar, inoculated into brain heart infusion broth, and incubated at 37°C overnight. Single rooted human mandibular premolars were sectioned below cementoenamel junction, enlarged, and vertically sectioned along the midsagittal plane. The samples were then placed in tissue culture wells inoculated with 2 ml of the bacterial solution and incubated at 37°C. MATERIALS AND METHODS: Group 1 E. cardamomum oil (cardamom), Group 2 T. ammi oil (ajwain), Group 3 5% sodium hypochlorite, and Group 4 Saline (control) (n = 10). At the end of the 2nd and 4th weeks, all groups were treated for 10 min with 3 ml of the respective solutions. Quantitative analysis was performed by serial dilution. RESULTS: T. ammi oil and sodium hypochlorite treated teeth showed complete elimination of both the 2-week-old and 4-week-old E. faecalis biofilm. Meanwhile, saline and E. cardamomum oil-treated teeth still showed the presence of E. faecalis. CONCLUSIONS: The use of T. ammi oil as a root canal irrigant solution can be considered as an alternative to sodium hypochlorite.

Visualizing mutagenic repair: novel insights into bacterial translesion synthesis.

DNA repair is essential for cell survival. In all domains of life, error-prone and error-free repair pathways ensure maintenance of genome integrity under stress. Mutagenic, low-fidelity repair mechanisms help avoid potential lethality associated with unrepaired damage, thus making them important for genome maintenance and, in some cases, the preferred mode of repair. However, cells carefully regulate pathway choice to restrict activity of these pathways to only certain conditions. One such repair mechanism is translesion synthesis (TLS), where a low-fidelity DNA polymerase is employed to synthesize across a lesion. In bacteria, TLS is a potent source of stress-induced mutagenesis, with potential implications in cellular adaptation as well as antibiotic resistance. Extensive genetic and biochemical studies, predominantly in Escherichia coli, have established a central role for TLS in bypassing bulky DNA lesions associated with ongoing replication, either at or behind the replication fork. More recently, imaging-based approaches have been applied to understand the molecular mechanisms of TLS and how its function is regulated. Together, these studies have highlighted replication-independent roles for TLS as well. In this review, we discuss the current status of research on bacterial TLS, with emphasis on recent insights gained mostly through microscopy at the single-cell and single-molecule level.

A comparative study of the evolution of cellobiose utilization in Escherichia coli and Shigella sonnei.

The chb operon of Escherichia coli is involved in the utilization of chitooligosaccharides. While acquisition of two classes of mutations leading to altered regulation of the chb operon is necessary to confer the ability to utilize the glucose disaccharide cellobiose to wild-type strains of E. coli, in the closely related organism Shigella sonnei, Cel+ mutants arise relatively faster, requiring only a single mutational event. In Type I mutants, the insertion of IS600 at -21 leads to ChbR regulator-independent, constitutive expression of the operon. In Type II mutants, the insertion of IS2/600 within the distal binding site of the negative regulator NagC leads to ChbR-dependent cellobiose-inducible expression of the operon. These studies underscore the significance of strain background, specifically the diversity of transposable elements, in the evolution of novel metabolic functions. Constitutive expression of the chb operon also enables utilization of the aromatic ß-glucosides arbutin and salicin, implying that the chb structural genes are inherently promiscuous.

Evaluation and comparison of the microhardness of enamel after bleaching with fluoride free and fluoride containing carbamide peroxide bleaching agents and post bleaching anticay application: An in vitro study.

AIMS AND OBJECTIVES: The purpose of the study was to evaluate and compare the microhardness of enamel after the application of anticay on bleached enamel with fluoride containing and fluoride free bleaching agent. MATERIALS AND METHODS: Twenty freshly extracted teeth decoronated and divided mesiodistally into two halves were randomly divided into five groups with 10 samples in each group. The enamel surface was treated as follows: Group 1 - no treatment, Group 2 - fluoride free bleaching agent, Group 3 - fluoride containing bleaching agent, and Group 4 - fluoride free bleaching agent followed by anticay application. The samples were subjected to indentation to test the microhardness using Vicker's hardness analyzer. CONCLUSION: Enamel microhardness significantly increased in samples where anticay was used after the application of bleaching agent.

Microleakage evaluation of Silorane-based composite and methacrylate-based composite in class II box preparations using two different layering techniques: an in vitro study.

AIM: The aim of this study was to compare the microleakage in Class II box preparations with the gingival margin above and below the cemento-enamel junction (CEJ) restored with Silorane composite and methacrylate composite using two different layering techniques. MATERIALS AND METHODS: Standardized box preparations (mesial box 1 mm above the CEJ and distal box 1 mm below the CEJ) were prepared in 60 upper premolars. The teeth were randomly divided into four groups containing 15 samples each; Group I: Restored with a Silorane composite using an oblique layering technique, Group II: Restored with Silorane composite using a vertical layering technique, Group III: Restored with methacrylate composite using the oblique layering technique, and Group IV: Restored with methacrylate composite using the vertical layering technique. The samples were stored in distilled water, followed by thermocycling and immersed in 2% methylene blue. The samples were sectioned and evaluated for microleakage at the gingival margin. STATISTICAL ANALYSIS: Kruskal-Wallis, Fischer exact test, Wilicoxon test, and Mann-Whitney U test. RESULTS: Silorane composite had significantly lesser microleakage. No significant difference in microleakage was observed above and below the CEJ for Silorane-based composite. CONCLUSION: Silorane composite resin showed lesser microleakage compared to methacrylate composite resin. CLINICAL SIGNIFICANCE: The Silorane-based composites improve the marginal adaptation due to their reduced shrinkage, thereby decreasing the residual stress at the adhesive-tooth interface.

Multifocal granular cell tumor presenting as an esophageal stricture.

INTRODUCTION: Granular cell tumors are uncommonly found in the gastrointestinal tract with slow progression and are usually benign though they may have propensity for malignant transformation. Initially attributed to neuronal origin through immunohistochemistry, there has been controversy with increasing reports of granular cell tumors of non-neural origin. CASE REPORT: We report a case of multifocal granular cell tumor involving the esophagus and stomach in a young female with history of dysphagia for 9 years with worsening symptoms. She had been managed at another facility with repeated dilations for presumed benign peptic stricture. Radial endosonography (EUS) of the proximal end of stricture showed a posterior submucosal esophageal mass that was heterogeneous and invaded into the muscularis propria. Fine-needle aspiration (FNA) showed large cells with granular cytoplasm along with spindle nuclei. Cells were initially checked for CD117 stain alone and found to be negative. A follow-up CT-guided core needle biopsy revealed similar granular cells that were positive for S-100. She underwent a two-stage transhiatal esophagogastrectomy as the tumor circumferentially involved the cervical esophagus and was adherent to the trachea and recurrent laryngeal nerve bilaterally. At surgery, there were two additional foci palpable in the proximal stomach. DISCUSSION AND CONCLUSION: As these tumors may have potential for malignant transformation and locoregional invasion, they should be considered while evaluating submucosal lesions of the esophagus even in young patients. A large number of granular cell tumors may be missed in the absence of S-100 staining, which should be requested when granular cells are seen on cytology obtained by EUS FNA as this can be a minimally invasive diagnostic modality for these tumors. Other foci should be sought at surgery as they have a propensity for locoregional spread.