Identifying major histocompatibility complex class II-DR molecules in bovine and swine peripheral blood monocyte-derived macrophages using mAb-L243.

Major histocompatibility complex class II (MHC-II) molecules are involved in immune responses against pathogens and vaccine candidates' immunogenicity. Immunopeptidomics for identifying cancer and infection-related antigens and epitopes have benefited from advances in immunopurification methods and mass spectrometry analysis. The mouse anti-MHC-II-DR monoclonal antibody L243 (mAb-L243) has been effective in recognising MHC-II-DR in both human and non-human primates. It has also been shown to cross-react with other animal species, although it has not been tested in livestock. This study used mAb-L243 to identify Staphylococcus aureus and Salmonella enterica serovar Typhimurium peptides binding to cattle and swine macrophage MHC-II-DR molecules using flow cytometry, mass spectrometry and two immunopurification techniques. Antibody cross-reactivity led to identifying expressed MHC-II-DR molecules, together with 10 Staphylococcus aureus peptides in cattle and 13 S. enterica serovar Typhimurium peptides in swine. Such data demonstrates that MHC-II-DR expression and immunocapture approaches using L243 mAb represents a viable strategy for flow cytometry and immunopeptidomics analysis of bovine and swine antigen-presenting cells.

Performance of posterior third-generation monolithic zirconia crowns in a complete digital workflow: A three-year prospective clinical study.

Background/purpose: Translucent monolithic zirconia restorations have recently introduced. The purpose of this study was to evaluate the clinical behavior and the survival rate of the posterior third-generation monolithic zirconia crowns (MZCs) during three years of clinical service. Materials and methods: Twenty-four patients who needed thirty crowns were enrolled in this study. Digital impressions were made, and the crowns were milled and cemented with a resin cement. The crowns outcomes were assessed using the California Dental Association's (CDA) criteria. Gingival index (GI), plaque index (PI), and periodontal probing depth (PPD) for MZCs and contralateral natural teeth (control) were assessed. Margin index (MI) for MZCs was also assessed. Data analysis was conducted using the Wilcoxon signed-rank and the Friedman tests. Results: The 3-year survival rate was 100%. All MZCs were rated as satisfactory throughout the follow-up period, and no biological or mechanical complications were observed. No differences were recorded when GI, PI and PPD at 3-year of follow-up were compared to baseline. No differences were recorded between crowned and control teeth. The MI remained stable throughout the study period. Conclusion: The third-generation monolithic zirconia appears to be a good treatment option for the rehabilitation of posterior single teeth.

Biochemical analysis of Hyalomma dromedarii salivary glands and gut tissues using SR-FTIR micro-spectroscopy.

Ticks are obligatory voracious blood feeders infesting diverse vertebrate hosts, that have a crucial role in the transmission of diverse pathogens that threaten human and animal health. The continuous emergence of tick-borne diseases due to combined worldwide climatic changes, human activities, and acaricide-resistant tick strains, necessitates the development of novel ameliorative tick control strategies such as vaccines. The synchrotron-based Fourier transform infrared micro-spectroscopy (SR-FTIR) is a bioanalytical microprobe capable of exploring the molecular chemistry within microstructures at a cellular or subcellular level and is considered as a nondestructive analytical approach for biological specimens. In this study, SR-FTIR analysis was able to explore a qualitative and semi-quantitative biochemical composition of gut and salivary glands of Hyalomma dromedarii (H. dromedarii) tick detecting differences in the biochemical composition of both tissues. A notable observation regarding Amide I secondary structure protein profile was the higher ratio of aggregated strands in salivary gland and beta turns in gut tissues. Regarding the lipid profile, there was a higher intensity of lipid regions in gut tissue when compared to salivary glands. This detailed information on the biochemical compositions of tick tissues could assist in selecting vaccine and/or control candidates. Altogether, these findings confirmed SR-FTIR spectroscopy as a tool for detecting differences in the biochemical composition of H. dromedarii salivary glands and gut tissues. This approach could potentially be extended to the analysis of other ticks that are vectors of important diseases such as babesiosis and theileriosis.

Evaluation of Antimicrobial Properties, Cell Viability, and Metalloproteinase Activity of Bioceramic Endodontic Materials Used in Vital Pulp Therapy.

This study aimed to evaluate the antimicrobial properties, cell viability, and matrix metalloproteinase (MMP) inhibition capacity of several endodontic materials aimed at vital pulp therapy: Pro Root MTA®, EndoSequence®, Biodentine®, MTA Angelus®, TheraCal LC®, and BioC Repair®. The materials were prepared according to the manufacturer's instructions. Antimicrobial tests were conducted using a microcosm biofilm model, cell viability was assessed using murine fibroblasts (L929), and MMP activity was analyzed through electrophoresis. The results showed that BioC Repair®, Biodentine®, and EndoSequence® exhibited similar antimicrobial properties, while MTA Angelus® and ProRoot MTA® had inferior results but were comparable to each other. In terms of cell viability, no significant differences were observed among the materials. EndoSequence® demonstrated the highest MMP inhibition capacity. In conclusion, BioC Repair®, Biodentine®, EndoSequence®, and TheraCal® showed better antimicrobial properties among the tested materials. The materials did not exhibit significant differences in terms of cytotoxicity. However, EndoSequence® displayed superior MMP inhibition capacity.

Update on olfactory neuroblastoma.

Olfactory neuroblastomas are uncommon malignancies that arise from olfactory receptor cells located high in the nasal cavity. Accurate diagnosis plays a crucial role in determining clinical results and guiding treatment decisions. Diagnosis can be a major challenge for pathologists, especially when dealing with tumours with poor differentiation. The discovery of several molecular and immunohistochemical markers would help to overcome classification difficulties. Due to the paucity of large-scale studies, standardisation of diagnosis, treatment and prediction of outcome remains a challenge. Surgical resection by endoscopic techniques with the addition of postoperative irradiation is the treatment of choice. In addition, it is advisable to consider elective neck irradiation to minimise the risk of nodal recurrence. Molecular characterisation will help not only to make more accurate diagnoses but also to identify specific molecular targets that can be used to develop personalised treatment options tailored to each patient. The present review aims to summarise the current state of knowledge on histopathological diagnosis, the molecular biology and management of this disease.

In vitro characterization of a novel resin-based restorative material containing alkaline fillers.

OBJECTIVE: In this study, a comparative evaluation of the physicochemical properties of Cention N and other direct restorative materials was performed. Three restorative materials-a resin-modified glass ionomer (Fuji II LC), an alkasite-based resinous material (Cention N), and a resin composite (Tetric N Ceram)-were characterized in terms of degree of conversion, Knoop hardness number (KHN) ratio, flexural strength, elastic modulus, water sorption, water solubility, microshear bond strength to dentin, immediate microleakage, and radiopacity. METHODOLOGY: The microshear bond strength to dentin and microleakage of Cention N were evaluated with and without the application of an adhesive system (Tetric N Bond Universal). A one-way ANOVA test was used to analyze the data in terms of degree of conversion, KHN ratio, water sorption, water solubility, microshear bond strength to dentin, and radiopacity. A two-way ANOVA test (carried out considering the material type and ethanol aging as factors) was used to analyze the data in terms of flexural strength and elastic modulus. The Kruskal-Wallis test was used to statistically analyze the data on microleakage. A significance level of α=0.05 was used for all tests. RESULTS: Fuji II LC was found to have the highest degree of conversion, water sorption, and microleakage, as well as the lowest flexural strength. Cention N had the highest solubility; when used with an adhesive system, it achieved bond strength and microleakage similar to those of the Tetric N Ceram composite. Tetric N Ceram had the highest degree of conversion, KHN ratio, and radiopacity. Conclusion: The properties of Cention N validate its efficacy as an alternative direct restorative material when used in conjunction with an adhesive system.

Digital workflow for monolithic and veneered zirconia and metal-ceramic posterior fixed partial dentures: A five-year prospective randomized clinical trial.

Purpose To evaluate and compare the survival, success rates, and biological and technical complications of three-unit posterior monolithic and veneered zirconia and metal-ceramic (MC) posterior fixed partial dentures (FPDs) fabricated using a digital workflow and computer-aided design and computer-aided manufacturing (CAD/CAM) over a 5-year follow-up.Methods Ninety patients in need of three-unit posterior FPDs were randomized to receive monolithic zirconia (MZ), veneered zirconia (VZ), and MC restorations (n = 30 each). Teeth preparations were scanned using an intraoral scanner, and restorations were milled and cemented with resin cement. Clinical performance and periodontal parameters were assessed at baseline and yearly up to 5 years after insertion. Data analysis was performed using the Kaplan-Meier method, Friedman test, and Wilcoxon signed-rank test with Bonferroni correction and Mann-Whitney U test.Results The 5-year survival rates of the MZ, VZ, and MC FPDs were 87%, 97%, and 100%, respectively (P = 0.04). Most complications were biological in nature. Only one MZ FPD fractured 58 months after placement. All the restorations were assessed as satisfactory at each recall. Differences were found in the gingival index score over time in the VZ and MC groups. The margin index remained stable throughout the follow-up period in both zirconia groups.Conclusions The results of this study suggest that using a digital workflow to fabricate posterior FPDs is an adequate treatment option and that monolithic zirconia could be a viable alternative to metal-ceramic or veneered zirconia. However, further long-term studies are necessary to provide stronger evidence in patients with bruxism.

Exact confidence intervals for population growth rate, longevity and generation time.

By quantifying key life history parameters in populations, such as growth rate, longevity, and generation time, researchers and administrators can obtain valuable insights into its dynamics. Although point estimates of demographic parameters have been available since the inception of demography as a scientific discipline, the construction of confidence intervals has typically relied on approximations through series expansions or computationally intensive techniques. This study introduces the first mathematical expression for calculating confidence intervals for the aforementioned life history traits when individuals are unidentifiable and data are presented as a life table. The key finding is the accurate estimation of the confidence interval for r, the instantaneous growth rate, which is tested using Monte Carlo simulations with four arbitrary discrete distributions. In comparison to the bootstrap method, the proposed interval construction method proves more efficient, particularly for experiments with a total offspring size below 400. We discuss handling cases where data are organized in extended life tables or as a matrix of vital rates. We have developed and provided accompanying code to facilitate these computations.

In vitro characterization of a novel resin-based restorative material containing alkaline fillers

Abstract Objective: In this study, a comparative evaluation of the physicochemical properties of Cention N and other direct restorative materials was performed. Three restorative materials—a resin-modified glass ionomer (Fuji II LC), an alkasite-based resinous material (Cention N), and a resin composite (Tetric N Ceram)—were characterized in terms of degree of conversion, Knoop hardness number (KHN) ratio, flexural strength, elastic modulus, water sorption, water solubility, microshear bond strength to dentin, immediate microleakage, and radiopacity. Methodology: The microshear bond strength to dentin and microleakage of Cention N were evaluated with and without the application of an adhesive system (Tetric N Bond Universal). A one-way ANOVA test was used to analyze the data in terms of degree of conversion, KHN ratio, water sorption, water solubility, microshear bond strength to dentin, and radiopacity. A two-way ANOVA test (carried out considering the material type and ethanol aging as factors) was used to analyze the data in terms of flexural strength and elastic modulus. The Kruskal-Wallis test was used to statistically analyze the data on microleakage. A significance level of α=0.05 was used for all tests. Results: Fuji II LC was found to have the highest degree of conversion, water sorption, and microleakage, as well as the lowest flexural strength. Cention N had the highest solubility; when used with an adhesive system, it achieved bond strength and microleakage similar to those of the Tetric N Ceram composite. Tetric N Ceram had the highest degree of conversion, KHN ratio, and radiopacity. Conclusion: The properties of Cention N validate its efficacy as an alternative direct restorative material when used in conjunction with an adhesive system.

Structural definition of babesial RAP-1 proteins identifies a novel protein superfamily across Apicomplexa.

Apicomplexan protozoa are intracellular parasites of medical and economic importance. These parasites contain specialized apical complex organelles, including rhoptries, that participate in the process of host cell invasion. Conserved antigens expressed in the rhoptries are rational vaccine targets, but whether conservation of protein structure is a functional requirement for invasion remains unknown. Novel protein structural modeling enables identification of structurally conserved protein families that are not evident by sequence analysis alone. Here we show by AlphaFold2 structural modeling that the rhoptry-associated protein 1 superfamily of the Piroplasmida hemoparasites Babesia and Theileria (pRAP-1) is structurally conserved, with the core conserved region being composed of a globin-like and a 4-helix bundle subdomain. Search for structurally related members of this protein family in other apicomplexan parasites revealed structural homologues of pRAP-1 in several species of Plasmodium, Toxoplasma gondii and other members of the Sarcocystidae family. Based on these structural findings, pRAP-1 is a conserved apical complex protein, but whether these proteins share functional features in different species remains unknown. Identification of widely conserved elements involved in infection in these parasites will enhance our knowledge of invasion mechanisms, and facilitate the design of methods for controlling diseases that affect humans and animals globally.

Diabetic Muscle Infarction: An Uncommon Diabetic Complication With a Lack of Standardized Treatment.

Diabetic muscle infarction (DMI) is a rare yet serious complication that has been strongly associated with uncontrolled diabetes, although other risk factors are unclear. DMI is an uncommon complication of diabetes with a lack of structured guidelines for evaluation or management. End-stage renal disease (ESRD) could have further implications in patients with DMI in terms of management given that nonsteroidal anti-inflammatory drugs (NSAIDs), which have been shown to reduce the recovery times and recurrence of DMI, could be contraindicated. We present a rare case of DMI in an African American man with ESRD who presented for new-onset right lower-extremity pain and swelling. We discuss the challenges involved with the diagnosis and treatment of this rare condition. This case adds to the knowledge of DMI, which is limited because of the low incidence of this condition, and it helps us understand how this condition affects the African American population and patients with ESRD.

Trifluoromethylarylation of alkenes using anilines.

Nitrogen containing compounds, such as anilines, are some of the most widespread and useful chemical species, although their high and unselective reactivity has prevented their incorporation into many interesting transformations, such as the functionalization of alkenes. Herein we report a method that allows the trifluoromethylarylation of alkenes using anilines, for the first time, with no need for additives, transition metals, photocatalysts or an excess of reagents. An in-depth mechanistic study reveals the key role of hexafluoroisopropanol (HFIP) as a unique solvent, establishing a hydrogen bonding network with aniline and trifluoromethyl reagent, that is responsible for the altered reactivity and exquisite selectivity. This work uncovers a new mode of reactivity that involves the use of abundant anilines as a non-prefunctionalized aromatic source and the simultaneous activation of trifluoromethyl hypervalent iodine reagent.

Diagnostic impact of RNA-based next-generation sequencing fusion panel for solid tumors: A single-institution experience.

OBJECTIVES: Gene rearrangements frequently act as oncogenic driver mutations and determine the onset and progression of cancer. RNA-based next-generation sequencing (NGS) is being used with increasing frequency for solid tumors. The purpose of our study is to investigate the feasibility and utility of an RNA-based NGS fusion panel for solid tumors. METHODS: We conducted a retrospective, single-institution review of fusion panels requested between May 2022 and March 2023. Demographic, clinical, pathologic, and molecular findings of the patients were reviewed. The utility of the RNA-based NGS fusion panel for the pathologic diagnosis of solid tumors was assessed. RESULTS: Our study included 345 cases, and a fusion event was identified in 24.3% (78/321) of cases. Among the 110 cases submitted for diagnostic purposes, a fusion event was detected in 42.7% (47/110) of cases. The results led to refinement or clarification of the initial diagnosis in 31.9% (15/47) of cases and agreement or support for the initial diagnosis in 59.6% (28/47) of cases. Furthermore, our study indicated that the overall cellularity (tumor and normal tissue) of the tested specimen influences the success of the testing process. CONCLUSIONS: In summary, this study demonstrated the feasibility and utility of an RNA-based NGS fusion panel for a wide variety of solid tumors in the appropriate clinicopathologic context. These findings warrant further validation in larger studies involving multiple institutional patient cohorts.

Impact of Equine and Camel Piroplasmosis in Egypt: How Much Do We Know about the Current Situation?

Piroplasmosis is a global tick-borne disease caused by hemoprotozoan parasites, which causes high morbidity and substantial economic losses in farm animals. Equine and camel piroplasmosis causes significant losses worldwide and in Egypt. The multifactorial effects and overall impact of equine and camel piroplasmosis in Egypt remain poorly characterized. However, several Babesia and Theileria spp. as well as potential tick vectors affecting these two species have been identified in the country. Equine and camel piroplasmosis has been reported by all governates in the country. Thus, in this work, we intend to provide a broad depiction of the current approaches used for diagnosis and control and the impact of piroplasmosis on the equine and camel industries in Egypt. We also identified current gaps in knowledge that might help develop future research efforts towards improved intervention and control of equine and camel piroplasmosis. It is important to develop specific diagnostic tools suitable for the early and chronic diagnosis of this disease. Altogether, the current situation warrants the development of large-scale epidemiological studies in order to obtain an accurate estimate for equine and camel piroplasmosis to secure the highly needed food resources in the country.

Development of Lead-Free Radiation Shielding Material Utilizing Barium Sulfate and Magnesium Oxide as Fillers in Addition Cure Liquid Silicone Rubber.

The radiological protection has the purpose of safeguarding the physical well-being of the user, preventing exposure to detrimental levels of ionizing radiation. This study introduces a novel, cost-effective category of lead-free elastomeric material designed for radiation shielding. The filler compounds utilized are notably lighter than conventional lead-based materials, enhancing user ergonomics during application. They comprise of a blend of barium sulfate combined or not with magnesium oxide with addition-cure liquid silicone rubber. To ensure the effectiveness of the radiation shielding, X-ray transmission measurements were performed for the different thicknesses of the materials and the results compared with Monte Carlo simulations. Additionally, the physical properties of the new materials, such as density, homogeneity, tensile strength, viscosity, and wettability, were also evaluated. The findings indicate that both materials fulfill the requirement for application in radiation protection garments.

Experimental infection of non-immunosuppressed and immunosuppressed goats reveals differential pathogenesis of Babesia aktasi n. sp.

Babesiosis is an acute and persistent tick-borne disease caused by protozoan parasites of the genus Babesia. These hemoparasites affect vertebrates globally, resulting in symptoms such as high fever, anemia, jaundice, and even death. Advancements in molecular parasitology revealed new Babesia species/genotypes affecting sheep and goats, including Babesia aktasi n. sp., which is highly prevalent in goats from Turkiye's Mediterranean region. The objective of this study was to investigate the pathogenesis of B. aktasi infection in immunosuppressed (n=7) and non-immunosuppressed (n=6) goats. These animals were experimentally infected with fresh B. aktasi infected blood, and their clinical signs, hematological and serum biochemical parameters were monitored throughout the infection. The presence of parasites in the blood of immunosuppressed goats was detected by microscopic examination between 4 and 6 days after infection, accompanied by fever and increasing parasitemia. Goats that succumbed acute disease exhibited severe clinical signs, such as anemia, hemoglobinuria, and loss of appetite. However, the goats that survived showed milder clinical signs. In the non-immunosuppressed group, piroplasm forms of B. aktasi were observed in the blood within 2-5 days after inoculation, but with low (0.01-0.2%) parasitemia. Although these goats showed loss of appetite, typical signs of babesiosis were absent except for increased body temperature. Hematological analysis revealed significant decreases in the levels of red blood cells, leukocytes and platelet values post-infection in immunosuppressed goats, while no significant hematological changes were observed in non-immunosuppressed goats. In addition, serum biochemical analysis showed elevated transaminase liver enzymes levels, decreased glucose, and lower total protein values in the immunosuppressed group post-infection. Babesia aktasi, caused mild disease with minor clinical symptoms in non-immunosuppressed goats. However, in immunosuppressed goats, it exhibited remarkable pathogenicity, leading to severe clinical infections and death. In conclusion, this study provides valuable insights into the pathogenicity of the parasite and will serve as a foundation for future research aimed at developing effective prevention and control strategies against babesiosis in small ruminants. Further research is required to investigate the pathogenicity of B. aktasi in various goat breeds, other potential hosts, the vector ticks involved, and its presence in natural reservoirs.

Time to diagnosis in rapid exome/genome sequencing in the clinical inpatient setting.

Exome and genome sequencing are clinically available, with many laboratories offering expedited testing (e.g., "rapid" and "ultra-rapid"). With the increase in uptake of expedited testing, there is a need for the development of inpatient protocols for best practices based on real-life data. A retrospective 2-year review (October 2019-November 2021) of the utilization of rapid exome and genome sequencing for inpatient cases at a tertiary care center using a utilization management tracking database with subsequent chart review was performed. Thirty-three expedited "rapid/priority" exome/genome tests were performed clinically. The average total turnaround time (TAT) was 17.88 days (5-43 days) with an average TAT of 13.97 days (3-41 days) for the performing laboratory. There were 5 positive diagnostic results (15.2%), 3 likely positive diagnostic results (9%), 2 noncontributory results (6%), and 26 nondiagnostic results (69.7%). Real-life data suggest that there is an approximately 3.91-day lag in getting samples to the performing laboratory. Although laboratories may advertise their expected TAT, a number of factors can potentially impact the actual time from test order placement to communication of the results for clinical use. Understanding the points of delay will enable the development of internal protocols and policies to improve time to diagnosis.

Effect of Modified Triple-Layer Application on the Bond Strength of Different Dental Adhesive Systems to Dentin.

The goal of this article was to assess the effect of modified triple-layer application (MTLA) in conjunction with the active bonding technique on the bond strength of four adhesive systems to dentinal substrate. The adhesives tested were Prime&Bond Universal (PBU), OptiBond Universal (OBU), OptiBond FL (OBFL), and Clearfil SE (CSE). The adhesives were applied according to the following strategies: single active application (A) and triple adhesive layer application including Active-Passive-Passive (APP); AAP; and AAA. The micro-tensile bond strength test was evaluated following 24 h or 6 months of storage. The composite-dentin interface morphology was investigated using scanning electron microscopy. The data were statistically analyzed with a significance level of α = 0.05. At 24 h of aging, all of the factors tested were not significant (p > 0.05) for CSE. For OBFL, OBU, and PBU, statistically higher values were observed for the A technique (p < 0.05). Plus, there were no significant variances between the APP, AAP, and AAA techniques (p > 0.05) for OBFL and PBU. However, for OBU, there were no significant differences between the A and AAA techniques (p > 0.05). After 6 months of aging, the A technique showed statistically higher values when compared to the other techniques (p < 0.01), except for OBFL, where the A and AAA techniques showed promising outcomes. When comparing the bond strength values of 24 h and 6 months, only for PBU, all of the techniques used resulted in bond strength stability over time (p > 0.05). Thicker adhesive layers were observed when MTLA was applied. Only the OBFL adhesive showed the formation of resin tags in all of the modalities tested. The bonding performances of the different application techniques used were material-dependent.

Effectiveness of Different Application Modalities on the Bond Performance of Four Polymeric Adhesive Systems to Dentin.

One of the major goals of adhesive dentistry is to improve the interaction of the already-existing adhesives with different substrates by using different application techniques. Thus, the objective of the present in vitro study was to assess the bond performance of four adhesive systems, Prime&Bond Universal (PBU), Clearfil SE Bond (CSE), OptiBond Universal (OBU), and OptiBond FL (OBFL), to dentin using various application modes: passive application (PA), active application (AA), Compo-Vibes modified application (CVM), and Compo-Vibes application (CV). Eighty extracted human molars were allocated into four groups based on the application modalities tested. The micro-tensile bond strength as well as fracture mode were tested in accordance with ISO/TS 11.405 after 24 h and 6 months of aging. Adhesive contact angle (CA) and scanning electron microscope analysis were also performed (n = 3). Statistical tests were performed with α = 0.05. After 24 h, a significant difference with a higher bond strength value was found for PBU in the AA modality and for CSE in the CVM modality (p < 0.05). However, no significant difference was shown between the techniques used among the other adhesives (OBFL and OBU). Moreover, at 24 h, only the PA demonstrated significant differences between the tested materials (p < 0.05). After 6 months, CSE, PBU, and OBU demonstrated significant differences between the techniques (p < 0.05), with a higher bond strength for CSE in AA and CVM modalities, for PBU in AA modality, and for OBU in AA and PA modalities. No significant differences were found between the techniques used among the OBFL (p > 0.05). In addition, only the CVM technique demonstrated significant differences between the tested materials after 6 months. CV and CVM showed a decreased value after aging for CSE and PBU, respectively. However, all the modalities decreased for OBU and OBFL after aging. All the adhesives showed marked resin infiltration into dentinal tubules in AA among all the modalities tested. Both universal adhesive systems (OBU and PBU) demonstrated statistically lower CA when compared to the other systems (CSE and OBFL) (p < 0.05) when applied in the PA mode. Concerning the AA mode, only CSE and OBFL were tested. The AA demonstrated lower CA values compared to the same adhesives in PA (p < 0.05). It could be concluded that the bond strength could be influenced by both materials and application techniques. It seems that the AA technique could be recommended as a gold standard for the application of an adhesive system to dentin. Plus, the CV and CVM modalities after 6 months of aging were considered stable for PBU and CSE, respectively. Consequently, the performance of these adhesive systems might vary when applied to other modalities. Future studies are needed to test this hypothesis.