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Cardiovascular diseases, resulting from the deposition of clots in blood vessels, are the leading cause of death worldwide. Fibrinolytic enzymatic activity can catalyze blood clot degradation. Findings show that 36 fungal isolates recovered from Caatinga soils have the potential to produce fibrinolytic protease under submerged conditions. About 58 % of the isolates displayed fibrinolytic activity above 100 U/mL, with Mucor subtilissimus UCP 1262 being the most active. The protease was biochemically and biophysically characterized, showing that the enzyme had a high affinity for SAApNA substrate and was significantly inhibited by fluoride methyl phenyl sulfonyl-C7H7FO2S, suggesting that it is a chymotrypsin-like serine protease. The highest enzyme activity was detected at pH 5.0 and 28 °C. This fibrinolytic protease's far-UV circular dichroism (CD) showed that its secondary structure was primarily α-helical. The purified fibrinolytic enzyme may represent a novel therapeutic agent for treating thrombosis. At temperatures above 65 °C, the enzyme lost all its secondary structure. Its melting temperature was 58.1 °C, the denaturation enthalpy 85.1 kcal/mol, and the denaturation entropy 0.26 kcal/K∙mol.

Objectives: This study aimed to assess the surface roughness, surface free energy (SFE), and shear bond strength (SBS) on a lithium disilicate glass-ceramic surface following varying etching protocols (time variation) and application of silane either with or without adhesive material., Methods and Materials: Lithium disilicate glassceramic (LDGC) computer-aided design and computer-aided manufacture (CAD/CAM) blocks were cut using a slow-speed cutting mechanism. CAD/CAM blocks were then evaluated for surface roughness, 6 groups (n=20); SFE,12 groups (n=5); and SBS, 10 groups (n=10). The cut CAD/CAM blocks were randomly allocated to 28 groups. Groups were based on the following: 30 or 90 seconds of etching with 9% hydrofluoric acid (HF); application or absence of silane coupling agent (Sil); and application or absence of adhesive (Adh).The control group (Cont) had untreated surfaces. Unetched surfaces were surveyed with only silane (Sil), only adhesive (Adh), or silane+adhesive (SilAdh). Further etched groups were HF30 with HF for 30 seconds, HF30-Sil, HF30-Adh, and HF30-SilAdh. Alternative 90-second etching times produced similar groups: HF90, HF90-Sil, HF90-Adh, and HF90-SilAdh. A digital profilometer was used to assess the surface roughness of specimens, and two readings were recorded. Sessile drop analysis was used to examine SFE specimens, and the OWRK model was modified to measure liquid surface tension. A universal testing machine (UltraTester, Ultradent Products, Inc, South Jordan, UT, USA) was utilized for the SBS test, with the crosshead speed set at 0.5 mm/min until failure. Representative treated specimens from each group were submitted to surface morphological evaluation and chemical analysis using scanning electron microscopy/energy dispersive x-ray spectroscopy (SEM/EDXS) (n=3). After data collection, evaluation using one- or two-way analysis of variance and the post-hoc Tukey test (α=5%) was conducted., Results: A longer etching time of 90 seconds produced a rougher surface. After the 90-second etching process, SFE displayed the greatest values; nevertheless, the use of silane did not affect SFE. For every group examined, the application of silane followed by adhesive resulted in an increase in SBS and more stable bonding over time. SEM/EDXS showed that etching times did affect the amount of cerium on the surface and altered surface morphology., Conclusions: Higher and more consistent bond strengths have been observed with longer etching periods. Silane and adhesive application on the ceramic surface showed stronger and enhanced bond strength, specifically when longer etching times were employed., (©Operative Dentistry, 2024.)

Purpose: This systematic review and meta-analysis aimed to evaluate the depth distortion and angular deviation of fully-guided tooth-supported static surgical guides (FTSG) in partially edentulous arches compared to partially guided surgical guides or freehand., Material and Methods: This study followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines and was registered in the Open Science Framework (OSF). The formulated population, intervention, comparison, and outcome (PICO) question was: "In partially edentulous arches, what are the depth distortion and angular deviation of FTSG compared to partially guided surgical guides or freehand?" The search strategy involved four main electronic databases, and an additional manual search was completed in November 2023 by following an established search strategy. Initial inclusion was based on titles and abstracts, followed by a detailed review of selected studies, and clinical studies that evaluated the angular deviations or depth distortion in FTSG in partial arches, compared to partially guided surgical guides or freehand, were included. In FTSG, two surgical approaches were compared: open flap and flapless techniques, and two digital methods were assessed for surgical guide design with fiducial markers or dental surfaces. A qualitative analysis for clinical studies was used to assess the risk of bias. The certainty of the evidence was assessed according to the grading of recommendations, assessment, development, and evaluations (GRADE) system. In addition, a single-arm meta-analysis of proportion was performed to evaluate the angular deviation of freehand and FTSG., Results: Ten studies, published between 2018 and 2023, met the eligibility criteria. Among them, 10 studies reported angular deviations ranging from -0.32° to 4.96° for FTSG. Regarding FTSG surgical approaches, seven studies examined the open flap technique for FTSG, reporting mean angular deviations ranging from 2.03° to 4.23°, and four studies evaluated flapless FTSG, reporting angular deviations ranging from -0.32° to 3.38°. Six studies assessed the freehand surgical approach, reporting angular deviations ranging from 1.40° to 7.36°. The mean depth distortion ranged between 0.19 mm to 2.05 mm for open flap FTSG, and between 0.15 mm to 0.45 mm for flapless FTSG. For partially guided surgical guides, two studies reported angular deviations ranging from 0.59° to 3.44°. Seven studies were eligible for meta-analysis, focusing on the FTSG in open flap technique, with high heterogeneity (I 2 (95%CI) = 92.3% (88.7%-96.4%)). In contrast, heterogeneity was low in studies comparing freehand versus FTSG in open flap techniques (I 2 (95%CI) = 21.3% (0.0%-67.8%)), favoring the FTSG surgical approach., Conclusion: In partially edentulous arches, FTSG systems exhibited less angular deviation than freehand and partially guided surgical guides. Flapless surgical approaches were associated with reduced angular deviation and depth distortion, suggesting a potential preference for the FTSG method in these procedures., (© 2024 The Author(s). Journal of Prosthodontics published by Wiley Periodicals LLC on behalf of American College of Prosthodontists.)

Objectives: To assess the impact of elastic gradients formed among restorative material, cement, and substrate on the fracture resistance of tri-layer restorative systems., Methods: Four CAD/CAM materials were utilized, two glass-ceramics (IPS e.max CAD, Vita Suprinity) and two resin-ceramic hybrids (Vita Enamic, Lava Ultimate). Their fracture resistance was examined by biaxial flexure (n = 8) and Hertzian indentation (n = 10) tests. Statistical analysis was conducted using ANOVA and Tukey tests (p = 5 %). Finite element analysis (FEA) was employed to simulate the Hertzian indentation test and elucidate the stress-fields formed on the intaglio surface below the loading area., Results: The biaxial flexural strength (MPa) of glass-ceramics exceeded the hybrid materials (e.max 417 a , Suprinity 230 b , Enamic 138 c , and Lava Ultimate 183 bc ). Conversely, the load-bearing capacity (N) of the materials bonded to dentin analog demonstrated the opposite trend, with the hybrid materials achieving superior results (e.max 830  C , Suprinity 660 D , Enamic 1822 B , and Lava Ultimate 2593  A ). The stress-fields observed by FEA were coherent with the experimental results for Hertzian flexural stresses (MPa): e.max 501  A , Suprinity 342  C , Enamic 406 B , whereas no tensile stress was observed at the intaglio surface of Lava Ultimate., Significance: Detailed analysis of the fracture resistance of the tri-layer restorative systems showed that the elastic gradients play a more significant role than the flexural strength of the restorative materials. The coherence of the elastic moduli between the restorative material and supporting structures results in reduced tensile stress concentration at the intaglio surface beneath the loading area and enhances the ability to withstand load., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The present study focused on evaluating the sequential fermentation (SF) method for FFase production from Aspergillus tamarii URM4634 using soybean bran as substrate. The SF was performed using soybean bran as substrate at 72 h and 30 °C and the maximum hydrolytic activity (44.00 U mL -1 ), corresponding to an increase of 2.98-fold to about SmF using sucrose as substrate. Already the maximum transfructosylating activity was 26.10 U mL -1 . The FFase presents maximum hydrolytic activity at pH 5.0-6.0 and transfructosylating at pH 6.0 and 60 °C for both enzyme activities. The enzyme showed a typical hydrolytic kinetic profile evidenced by more affinity by sucrose hydrolysis reaction than the fructosyl transfer one. From kinetic and thermodynamic data of thermal denaturation, it was observed that the enzyme presents suitable at 55 °C, evidenced by the large half-life (990.21 min) and D values (3289.41 min). The maximum release of reducing sugars (8.45 g L -1 ) was obtained in hydrolysis of 20% sucrose during 180 min. The results obtained for FFase production by SF proved that this method can be used satisfactorily for sucrose-degrading enzymes and can contribute to the development of the SF technique to produce different industrial-interest enzymes., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© King Abdulaziz City for Science and Technology 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

This study aimed to produce, characterize and purify a protease from Aspergillus heteromorphus URM0269. After production by solid fermentation of wheat bran performed according to a central composite design, protease was characterized in terms of biochemical, kinetic, and thermodynamic parameters for further purification by chromatography. Proteolytic activity achieved a maximum value of 57.43 U/mL using 7.8 g of wheat bran with 40 % moisture. Protease displayed high stability in the pH and temperature ranges of 5.0-10.0 and 20-30 °C, respectively, and acted optimally at pH 7.0 and 50 °C. The enzyme, characterized as a serine protease, followed Michaelis-Menten kinetics with a maximum reaction rate of 140.0 U/mL and Michaelis constant of 11.6 mg/mL. Thermodynamic activation parameters, namely activation Gibbs free energy (69.79 kJ/mol), enthalpy (5.86 kJ/mol), and entropy (-214.39 J/mol.K) of the hydrolysis reaction, corroborated with kinetic modeling showing high affinity for azocasein. However, thermodynamic parameters suggested a reversible mechanism of unfolding. Purification by chromatography yielded a protease purification factor of 7.2, and SDS-PAGE revealed one protein band with a molecular mass of 14.7 kDa. Circular dichroism demonstrated a secondary structure made up of 45.6 % α-helices. These results show the great potential of this protease for future use in the industrial area., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Collagenolytic proteases produced by Aspergillus heteromorphus URM0269 were extracted using a PEG/sulfate aqueous two-phase system (ATPS). A 2 3 factorial design was performed to analyze the independent variables: PEG molar mass (M PEG ), PEG concentration (C PEG ), and sulfate concentration (C sulf ). The extracted proteases were also evaluated for their optimum pH and stability at different pH levels (4.0 - 11.0) after 20 h of incubation. Collagen was extracted from mutton snapper ( Lutjanus analis) skin using acetic acid (0.5 mol L -1 ). The enzyme was preferentially partitioned to the PEG-rich phase (K > 1), whose highest purification factor and recovery (PF = 6.256 and Y = 404.432%) were obtained under specific conditions: M PEG 8000 g.mol -1 , C PEG 30%, C sulf 10%. The ATPS extraction provided an enzymatic activity range of pH 7.0 - 11.0, exhibiting greater stability compared to the crude extract. Approximately 80% of protease activity was maintained after 20 hours of incubation at all analyzed pH levels, except pH 11.0. Collagen extraction from L. analis skin yielded 8.056%, and both crude extract samples and ATPS-derived samples successfully hydrolyzed the extracted collagen, reaching peak hydrolysis after 36 hours of treatment. These findings demonstrate the feasibility of extracting highly purified and active proteases capable of hydrolyzing L. analis collagen.

Foals require maternal colostrum in the first hours of life to prevent failure of transfer of passive immunity (FTIP). Innovative storage methods such as lyophilization may enable conservation of colostrum immunoglobulins by a differentiated process of dehydration. The current study aimed to compare the quality of equine colostrum after freezing and after the lyophilization process. Thirty-one pregnant Quarter Horse mares were used. The IgG concentration of frozen and lyophilized colostrum was determined by simple radial immunodiffusion (SRID) and Brix refractometry. The physical-chemical composition (pH, total protein (TP), fat, lactose, salts, total solids (TS), and density) of the samples was evaluated and the lyophilized colostrum reconstitution test was performed. There were no significant differences (P > 0.05) in the variables IgG, fat, lactose, salts, TS, density, and pH between samples measured before and after lyophilization. There was a significant difference (P < 0.05) between the Brix average and the TP of the frozen and lyophilized colostrum samples. Lyophilization resulted in a small reduction (6.55%) in the IgG concentration measured by SRID. A strong positive correlation was observed between colostrum density and IgG concentration by SRID (r = 0.76) and between Brix and IgG concentration by SRID (r = 0.77). In the reconstitution test, the lyophilized colostrum was easily rehydrated in water, with full dilution, and remained stable. Lyophilization could be an alternative for the conservation of mare colostrum, since it is a very efficient process for retaining the physicochemical characteristics of the product, with minimal loss, particularly of IgG., Competing Interests: Declaration of Competing Interest None of the authors has any financial or personal relationships that could inappropriately influence or bias the content of the paper., (Copyright © 2023. Published by Elsevier Inc.)

Statement of Problem: Polymer-based computer-aided design and computer-aided manufacturing (CAD-CAM) materials have either a high content of ceramic filler particles or a ceramic network structure. Proper etching of these materials is challenging. Therefore, it is relevant to assess different surface etching strategies and morphological alterations as a result of these techniques., Purpose: The purpose of this 2-part in vitro study was to evaluate different etching strategies on the surface roughness and its relationship to the surface contact angle with a subsequent morphological characterization of different CAD-CAM materials., Material and Methods: Two CAD-CAM polymer-based materials and 1 CAD-CAM ceramic material were selected for this study. The materials were treated with different strategies, including a combination of airborne-particle abrasion and an application of 9% hydrofluoric acid etching. After the surface treatment, roughness measurements (Sa) were assessed by using an optical profilometer on an area of 1.47 mm 2 . Subsequently, the contact angle on the surface was examined by means of the sessile drop technique and an optical contact angle meter. The data were analyzed by using a 2-way analysis of variance, the post hoc Tukey test (α=.05), and the Pearson correlation coefficient., Results: The 3 materials tested showed an increase in surface roughness when treated with surface airborne-particle abrasion. However, the materials with polymer in their composition concomitantly increased the contact angle. More hydrophilic surfaces were observed when hydrofluoric acid was applied, regardless of the composition of the materials. The surface roughness presented a strong positive linear tendency in the surface treatments used., Conclusions: The application of airborne-particle abrasion on the surface of the polymer-based CAD-CAM materials increased the roughness and contact angle. However, the application of acid etching after the airborne-particle abrasion decreased the contact angle for polymer-based CAD-CAM materials, resulting in considerable enhancement of the surface quality for proper bonding., (Copyright © 2021 Editorial Council for The Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Inulinases are enzymes of great interest in the food industry, especially due to their application in the synthesis of fructose and fructo-oligosaccharides. Moreover, some inulinases ( I ) also present invertase activity ( S ), making them useful for sucrose hydrolysis processes. In the present study, the production of inulinase by Aspergillus niger URM5741 was evaluated and optimized using two statistical approaches. First, the composition of the cultivation medium was determined through a simplex centroid mixture design, followed by the selection of optimal fermentation conditions using the Box-Behnken design. Based on these experimental designs, the maximum activities of inulinase (16.68 U mL -1 ) and invertase (27.80 U mL -1 ) were achieved using a mixture of wheat, soy, and oat brans (5 g), along with 2.5% inulin and 40% moisture. The inulinase exhibited optimum temperature and pH of 60 °C and 4.0, respectively, displayed a high affinity for both substrates, as evidenced by very-low Michaelis constant values (1.07-1.54 mM). A relative thermostability was observed at 55-60 °C as indicated by half-lives values ( I: 169.06-137.27 min; S: 173.29-141.52 min) and D -values ( I: 561.61-456.00 min; S: 575.65-470.11 min) which were further confirmed by the high activation energy (123.01 and 143.29 kJ mol -1 ). The enzyme demonstrated favorable results in terms of inulin and sucrose hydrolysis, being a maximum release of reducing sugars of 6.04 and 15.80 g L -1 , respectively. These results indicate that the sequential statistical approach proved to be beneficial to produce inulinase by A. niger URM5741, with the obtained enzyme considered promising for long-term industrial applications., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© King Abdulaziz City for Science and Technology 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Statement of Problem: Bonding to recently launched polymer-based computer-aided design and computer-aided manufacturing (CAD-CAM) materials has been challenging. Evidence regarding etching strategies for dual-phase CAD-CAM materials is sparse, but adequate bonding is crucial for the clinical success and longevity of a restoration., Purpose: The purpose of this 2-part in vitro study was to evaluate and compare the effect of surface treatment strategies on the microshear bond strength and work of adhesion of polymer-based and ceramic materials. In addition, chemical elements present on the surface and the interface morphology after using those strategies were also assessed., Materials and Methods: Two CAD-CAM polymer and 1 CAD-CAM ceramic materials were selected for this in vitro study. The materials were subjected to different surface treatment strategies, including airborne-particle abrasion and the application of 9% hydrofluoric acid. Specimens were submitted to microshear bond strength before and after thermocycling, and the failure mode was classified. The work of adhesion was calculated based on the water-to-air surface tension of 72.8 mN.m -1 and the Young- Dupré equation. The surfaces were submitted to energy-dispersive X-ray spectroscopy, and the interfaces were analyzed using a scanning electron microscope. Data were subjected to 2-way ANOVA and the Tukey post hoc test (α=.05)., Results: The highest microshear bond strength means were observed for the polymer-based materials when hydrofluoric acid or airborne-particle abrasion was applied. The 3 materials tested showed a decrease in microshear bond strength after thermocycling, except for applying airborne-particle abrasion to 1 of the polymer-based material tested. The ceramic material tested showed a high microshear bond strength with the application of airborne-particle abrasion and hydrofluoric acid combined. The work of adhesion varied across the materials and presented high means when hydrofluoric acid was used., Conclusions: A combination of airborne-particle abrasion plus hydrofluoric acid should be considered for polymer-based or feldspathic ceramic CAD-CAM materials. In this in vitro study, both etching procedures combined produced higher bonding values for all materials tested., (Copyright © 2021 Editorial Council for The Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Objectives: Answer the PICO question: Do class I and II posterior restorations in permanent teeth placed with high-viscosity glass-ionomer cement (HV-GIC) fail more than composite resin (CR) restorations?, Data: The study was registered in the PROSPERO database (CRD42020138290). Randomized and controlled clinical trials, comparing the performance of HV-GIC and CR in load bearing cavities of posterior permanent teeth were included. Cochrane risk of bias tool and GRADE were used to assess the quality and certainty of the evidence. Meta-analyses were performed for clinical outcomes on USPHS and FDI criteria for 12-, 24- and 36-months follow-ups., Sources: PubMed, Scopus and Web of Science were last searched on April 2, 2022, without language or date restrictions. Reference lists of primary studies and their related article link in PubMed were manually searched., Study Selection: Ten studies were included, while data from 8 were used for the meta-analyses. A total of 849 HV-GIC and 800 CR restorations were followed. The primary outcome was the fracture/retention of the restoration, with a comparable performance for both materials on all follow-ups. The 36 months follow-up for class I restorations (longest) showed risk difference of -0,00 (95%CI -0,03 to 0,03; p = 0,98) and no heterogeneity (p = 0,98, I 2 =0%). The certainty of the evidence is moderate, as all included studies were at an uncertain risk of bias., Conclusions: HV-GIC and CR presented comparable clinical performance in posterior permanent teeth up to 36 months. HV-GIV wear in class I restorations followed by 24 months was the only poorer result compared to CR., Clinical Significance: Conservative load bearing cavities in permanent posterior teeth can be restored with HV-GIC with comparable clinical performance to CR expected at least up to 3 years. HV-GIC is a valuable direct restorative option for posterior teeth in high caries risk patients, in which CR is frequently associated with failure., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Antimicrobial resistance is a threat to public health. The emergence of antibiotic-resistant Staphylococcus aureus represents a priority for the implementation of preventive measures. The objective was to isolate S. aureus in humans, animals, and animal health care environment, and to characterize the genotypic and phenotypic profile of antimicrobial resistance in these isolates. We isolated S. aureus from staff, animals, and environment of a veterinary hospital, and identified their antimicrobial resistance profiles. Samples were collected from 20 humans, 13 animals, 14 surfaces, 8 mobile phones, and 7 veterinarians' stethoscopes by using sterile swabs. S. aureus was isolated by culturing on mannitol salt agar and preliminary identification was done by Gram staining and catalase test. Subsequently, a polymerase chain reaction was performed for species confirmation and investigating their antimicrobial-resistant genotypic profiles. Phenotypic profiles of resistant isolates were determined using the disk-diffusion technique. Ten S. aureus isolates were recovered from 5/20 humans (25%), it was also recovered from 2/13 animals (15.38%), including 1 dog and 1 cat, and from 1/14 of surfaces (7.14%). The oxacillin-susceptible mecA-positive Staphylococcus aureus phenotype was identified in a feline. Most of the isolates carried at least two resistance genes of different antimicrobial classes, with 90% (9/10) presenting the gene blaZ, with 10% (1/10) presenting the gene mecA, 20% (2/10) presenting tet38, 10% (1/10) presenting tetM, 90% (9/10) presenting norA, 50% (5/10) presenting norC, 10% (1/10) presenting ermA, and 60% (6/10) presenting ermB. In antibiograms, resistance to penicillin was identified in all the isolates, resistance to erythromycin was identified in 80% (8/10), and all the isolate's resistance to erythromycin presented erythromycin-induced resistance to clindamycin. Antimicrobial resistance in the veterinary hospital requires attention due to the risk of interspecies transmission, gene transfer between bacteria that colonize companion animals and humans and, can make antimicrobial therapy difficult., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

β-fructofuranosidases (FFases) are enzymes involved in sucrose hydrolysis and can be used in the production of invert sugar and fructo-oligosaccharides (FOS). This last is an important prebiotic extensively used in the food industry. In the present study, the FFase production by Aspergillus tamarii Kita UCP 1279 was assessed by solid-state fermentation using a mixture of wheat and soy brans as substrate. The FFase presents optimum pH and temperature at 5.0-7.0 and 60 °C, respectively. According to the kinetic/thermodynamic study, the FFase was relatively stable at 50 °C, a temperature frequently used in industrial FOS synthesis, using sucrose as substrate, evidenced by the parameters half-life (115.52 min) and D -value (383.76 min) and confirmed by thermodynamic parameters evaluated. The influence of static magnetic field with a 1450 G magnetic flux density presented a positive impact on FFase kinetic parameters evidenced by an increase of affinity of enzyme by substrate after exposition, observed by a decrease of 149.70 to 81.73 mM on K m . The results obtained indicate that FFases present suitable characteristics for further use in food industry applications. Moreover, the positive influence of a magnetic field is an indicator for further developments of bioprocesses with the presence of a magnetic field.

Objective: The aim of this study was to evaluate the effectiveness of cleansing protocols to remove sealer residues using different cleaning strategies and the effect of bond strength of two universal adhesives to dentin impregnated with epoxy resin-based sealer., Materials and Methods: Fifty bovine dentin specimens were impregnated with epoxy resin-based sealer containing calcium hydroxide (Sealer Plus) and submitted to cleansing protocols (n = 10): negative control (NC), positive control (PC), 95% ethanol (ET), xylol (XI), and amyl acetate (AA). Specimens were evaluated by scanning electron microscope (SEM). Other 100 specimens were submitted to the same protocols (n = 20). Each protocol was divided into subgroups (n = 10) according to the universal adhesive system used: Scotchbond Universal (SU); Ambar Universal (AU). Bond strength was evaluated by micro-shear bond test (μSBT). Persistence of residues data were assessed with Kruskal-Wallis and Dunn's tests. μSBT data were analyzed with two-way ANOVA (α = 5%)., Results: ET presented higher persistence of residues compared to AA and XI (p < 0.05). AA and XI were similar from each other (p > 0.05). AU and SU presented similar μSBT values, regardless of the cleansing solution (p > 0.05). SU-NC presented the highest μSBT among all conditions (p < 0.05)., Conclusion: The bond strength of universal adhesives was not affected by different amounts of sealer residues after different cleaning protocols., Clinical Significance: Non-polar substances such as amyl acetate and xylol are effective for removing residues from epoxy resin-based endodontic sealers., (© 2022 Wiley Periodicals LLC.)

Inulinases are enzymes involved in the hydrolysis of inulin, which can be used in the food industry to produce high-fructose syrups and fructo-oligosaccharides. For this purpose, different Aspergillus strains and substrates were tested for inulinase production by solid-state fermentation, among which Aspergillus terreus URM4658 grown on wheat bran showed the highest activity (15.08 U mL -1 ). The inulinase produced by this strain exhibited optimum activity at 60 °C and pH 4.0. A detailed kinetic/thermodynamic study was performed on the inulin hydrolysis reaction and enzyme thermal inactivation. Inulinase was shown to have a high affinity for substrate evidenced by very-low Michaelis constant values (0.78-2.02 mM), which together with a low activation energy (19.59 kJ mol -1 ), indicates good enzyme catalytic potential. Moreover, its long half-life ( t 1/2 = 519.86 min) and very high D -value (1726.94 min) at 60 °C suggested great thermostability, which was confirmed by the thermodynamic parameters of its thermal denaturation, namely the activation energy of thermal denaturation ( E* d = 182.18 kJ mol -1 ) and Gibbs free energy (106.18 ≤ Δ G * d ≤ 111.56 kJ mol -1 ). These results indicate that A. terreus URM4658 inulinase is a promising and efficient biocatalyst, which could be fruitfully exploited in long-term industrial applications.

This work aimed to compare the production of collagenolytic proteases produced by M. subtilissimus UCP1262 in submerged fermentation (SF) and solid-state fermentation (SSF) as well as extracting in aqueous two-phase system (ATPS). Collagenolytic protease production was performed in using MS-2 culture medium (SF) and soybean bran as substrate (SSF). Subsequently, the fermented liquid from both fermentations were used for the extraction of enzyme by ATPS, it was verified the influence of different variables from a factorial design 23. In SSF the highest protease and collagenolytic activities were achieved with 362.66 U/mL and 179.81 U/mL, respectively. When compared with SF (26.33 and 18.70 U/mL) higher values were obtained in the activities. The protease partitioning from SF and SSF in ATPS showed a similar profile showing higher affinity for the polymer rich phase. The highest value for the response variable purification factor (3.49) was obtained in the system using SSF. Thus, SSF shows promise as a bioprocess for extracellular production of collagenolytic proteases, using of soybean bran as substrate had used sustainable raw material, aiming application this possible enzyme in the treatment of burns and postoperative scarring.

The aim of this study was to evaluate the effects of cleaning protocols on the presence of residues, open dentinal tubules, and bond strength of a self-etch adhesive plus a dual-cure resin cement to dentin in the fiber-post space. The methods for cleaning utilized were conventional irrigation (CI), rotary brush with continuous rotary movement (RB), and continuous ultrasonic irrigation (CUI). Sixty bovine incisors were used for fiber-post space preparation. Thirty roots were divided into three groups (n = 10) according to the cleaning protocols and submitted to scanning electron microscope to evaluate presence of residues and relative quantity of open dentinal tubules on dentin surface. Similar protocols were performed in the other thirty roots (n = 10) using a self-etching adhesive system (Scotchbond Universal) and a dual-cure resin cement (Relyx Unicem 2). After 6 months cervical, middle, and apical thirds sections of fiber-post space were submitted to push-out testing. To data ANOVA and Tukey's test were employed (α = 5%). RB and CUI showed the lowest presence of residues on dentin surface (p = .015) and the highest bond strength values (p < .001), regardless of the dentin third. CUI presented the highest quantity of open dentinal tubules in middle and apical thirds (p < .001). CI presented the lowest bond strength value of resin cement, regardless of dentin third (p < .001). Mechanical cleaning protocols, such as RB and CUI, presented better surface cleaning capabilities and higher bond strength than CI with dual-cure resin cement employed. The use of a mechanical cleaning protocol seems to cause appropriate condition on the root dentin surface. Mechanical cleaning of root dentin provided greater bond strength of the self-etching system with dual resin cement. Conventional irrigation did not provide adequate root dentin cleaning. RESEARCH HIGHLIGHTS: • The use of a mechanical cleaning protocol seems to cause appropriate condition on the root dentin surface. • Mechanical cleaning of root dentin provided greater bond strength of the self-etching system with dual resin cement. • Conventional irrigation did not provide adequate root dentin cleaning., (© 2022 Wiley Periodicals LLC.)

Statement of Problem: The influence of different glaze firing protocols and cooling on the behavior of computer-aided design and computer-aided manufacturing (CAD-CAM) ceramics is unclear., Purpose: The purpose of this in vitro study was to investigate the influence of different glaze firing protocols on the abrasion resistance, surface roughness, microhardness, and brittleness index of CAD-CAM ceramics., Material and Methods: Rectangular specimens of IPS e.max CAD (MAX), IPS Empress CAD (PRE), and CEREC Blocs (CER) were obtained and divided into 5 groups according to the glaze firing protocol (n=8): control (CO), 1 conventional glaze firing (GF), 2 conventional glaze firings (GF 2 ), 1 extended glaze firing (EG), and 2 extended glaze firings (EG 2 ). The coefficient of friction (μ) was determined with the pin-on-disk test, and specimens were analyzed with an optical interferometer to calculate volume loss and roughness. Vickers microhardness and the brittleness index were obtained with a microdurometer. Microhardness data were evaluated by 2-way ANOVA and the Sidak tests (α=.05). Data from other tests were assessed with nonparametric ANOVA and Bonferroni tests (α=.05)., Results: Groups GF 2 , EG, and EG 2 of MAX and PRE and all groups of CER presented higher levels of coefficient of friction. Groups EG and EG 2 of MAX and CER showed high volume loss and surface roughness. Groups EG and EG 2 of MAX and PRE showed lower hardness, as did all groups of CER. MAX-CO and groups GF and EG of PRE showed a lower brittleness index (P<.05), while CER groups showed no significant differences (P>.05)., Conclusions: Ceramic specimens with conventional glaze firing presented less surface wear than those with extended glaze firing. The number of firings did not have a statistically significant influence., (Copyright © 2022 Editorial Council for The Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

β-fructofuranosidases (FFases) are enzymes involved in sucrose hydrolysis and fructo-oligosaccharides' production which are of great interest for the food industry. FFase from Aspergillus tamarii URM4634 was extracted using PEG/Phosphate Aqueous Biphasic Systems (ABS), and the impact of magnetic field on the extraction behavior was evaluated. A 2 4 -full experimental design was employed to study the influence of molar mass of PEG, concentrations of PEG and phosphate and pH on the selected response variables, i.e., partition coefficient ( K ), purification factor ( PF ), activity yield ( Y ) and selectivity ( S ). The influence of magnetic field during partition and NaCl concentration on the same responses was also studied. The best results of FFase extraction without magnetic field ( K  = 0.50, PF  = 4.05, Y  = 72.66% and S  = 0.06) were observed at pH 8.0 using 12.5% (w/w) PEG 400 and 25% (w/w) NaH 2 PO 4 /K 2 HPO 4 . Application of the magnetic field allowed improving the performance, with the best results being obtained at the longest distance between magnets (lowest magnetic field) and absence of NaCl ( K  = 0.93, PF  = 4.22, Y  = 83.79% and S  = 0.09). The outcomes obtained demonstrate that ABS combination with low intensity magnetic field can be used as an efficient FFase pre-purification method.

The present study evaluated the influence of the variables polyethylene glycol (PEG) molar mass, pH, PEG concentration and sodium citrate concentration in the integrated production of the protease from Aspergillus tamarii Kita UCP1279 by extractive fermentation, obtaining as a response the partition coefficient ( K ), activity yield ( Y ) and concentration factor ( CF ). The enzyme preferably partitioned to the top phase and obtained in the system formed by variables M PEG = 400 g mol -1 , C PEG = 20% (w w -1 ), and C CIT = 20% (w w -1 ) and pH 6, in this condition were obtained CF  = 1.90 and Y  = 79.90%. The protease showed stability at a temperature of 60 °C for 180 min, with optimum temperature 40 °C and pH 8.0. For the ions and inhibitors effects, the protease activity increased when exposed to Fe 2+ , Ca 2+ and Zn 2 + and inhibited by EDTA, being classified as metalloprotease. The kinetic parameters K m (35.63 mg mL -1 ) and V max (1.205 mg mL -1  min -1 ) were also estimated. Thus, the protease showed desirable characteristics that enable future industrial applications, especially, for beer industry.

Adhesive strategies were evaluated on the bond strength of bi-functional monomers bonded to endodontically-treated-dentin (ETD). Superficial dentin was removed on human molars. Teeth were immersed in 5 mL 2.5% NaOCl, followed by immersion in 5 mL 17% EDTA. Dentin surface impregnated with epoxy resin-based sealer was then divided four groups (n=10): Scotchbond Multi-Purpose (SBMP); Single Bond Universal (SBU); Optibond All-in-One (OPB); and Tetric-N-Bond Universal (TBU). After 24-h or 1-year-of-storage specimens were submitted to microtensile bond strength (µTBS) and failure classification. A confocal laser scanning microscope (CLSM) evaluated the hybrid layer formation. Two-way ANOVA and Tukey-HSD test were performed (α=5%). The µTBS did not present statistical differences among adhesive strategies after 24-h. Significant differences were found after 1-year-of-storage. CLSM analysis showed water infiltration and consequently degradation of the hybrid layer after 1-year-of-storage. The use of SBU universal adhesive on the self-etching mode on ETD produced more stable bond over the 1-year-of-storage.

Statement of Problem: The post space must be prepared with techniques that do not interfere with the bond strength. These techniques are related to the availability of opened dentinal tubules and incidence of residue among the dentin, the adhesive, and the cement. Nevertheless, the effect of different irrigants during endodontic instrumentation is unclear., Purpose: The purpose of this in vitro study was to assess the incidence of residue, the number of opened dentinal tubules, and the bond strength of the cementation system to root dentin of post space after 3 different irrigation protocols., Material and Methods: The roots of bovine incisors were obturated, prepared to receive a fiber post, and divided into 3 groups as per the irrigation protocol: without irrigation, with alternating irrigation, and with continuous irrigation. Thirty roots (n=10) were evaluated by using scanning electron microscopy to assess the incidence of residue (×100 magnification) and the number of opened dentinal tubules (×2000 magnification). The fiber posts were cemented, and the remaining specimens (n=10) were subjected to pushout testing and failure mode evaluation., Results: All protocols presented a similar incidence of residue, regardless of the post space third (P>.05). The irrigation protocol without irrigation had the lowest number of opened dentinal tubules in the cervical and middle thirds (P<.05). The number of opened dentinal tubules was similar in all protocols in the apical third of the post space (P>.05). The irrigation protocol without irrigation had the lowest bond strength values in the cervical and middle thirds (P<.05). The irrigation protocol with continuous irrigation had the highest bond strength values in the apical third (P<.05)., Conclusions: Post space preparation without irrigation negatively affected the bond strength of the resin cementation system to root dentin., (Copyright © 2020 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Copaiba oil is a natural product used by Amazonian populations and recognized for its medicinal properties because it has significant antimicrobial activity for several pathogenic microorganisms. The present work aimed to evaluate and characterize the effect of natural oil produced by copaiba - Copaifera multijuga against multiresistant isolates of bubaline mastitis. The nitrocefin test was performed with isolates of Staphylococcus aureus from bubaline mastitis, which were 100% positive for beta-lactamase enzyme detection. Minimum Bactericidal Concentration (MBC) of 25% to 3.12% was obtained for Enterococcus faecalis and Escherichia coli and 50% and 25% for S. aureus, but Klebsiella pneumoniae and Bacillus subtilis were resistant. MBC with 12.5% and 6.25% oil were obtained for most multiresistant bubaline mastitis isolates from the states of Pernambuco, Ceará, Bahia and Alagoas. The results demonstrated the great potential of using copaiba natural oil in the treatment of buffalo mastitis.

The industrial demand for proteolytic enzymes is stimulating the search for new enzyme sources. Fungal enzymes are preferred over bacterial enzymes, and more effective and easier to extract. The aim of this work was to evaluate the potential of protease production by solid state fermentation (SSF) of Mucor subtilissimus UCP 1262, evaluate different specific activities, purify and partially characterize the enzyme in terms of biochemical as to the optimal pH and temperature. Initially, the enzyme crude extract was screened for 3 different proteolytic activities, collagenolytic (161.4 U/mL), keratinolytic (39.6 U/mL) and fibrinolytic (26.1 U/mL) in addition to conventional proteinase activity. After ammonium sulfate precipitation, the active fractions with fibrinolytic activity were dialyzed in 15 mM Tris-HCl buffer, pH 8, loaded onto DEAE-Sephadex A50 ion-exchange column and gel filtrated through Superdex 75 HR10/300. The enzyme showed a fibrinolytic maximum activity at 40 C and pH 9,0. The purified enzyme showed activity against a chromogenic chymotrypsin substrate, SDS-PAGE showing a molecular mass of approximately 70 kDa and, the specific activity of 25.93 U/mg. These characteristics suggest that the enzyme could be and efficiently produced in a simple and low-cost way using Mucor subtilissimus UCP 1262 in SSF.

β-fructofuranosidase (FFase) from Aspergillus tamarii URM4634 was immobilized covalently in chitosan beads. It was characterized biochemically, studied in terms of kinetic and thermodynamic parameters, and applied on conversion of sucrose for invert sugar production in a packed bed reactor (PBR). The optimum reactional conditions were determined and obtained at pH 5.0 and 60 °C. FFase was thermostable at 50-55°C. At 50°C, the enzyme shows longer half-life (t 1/2 ) (594.13 min) and a higher D-value (1,973.64 min). This indicates that immobilized FFase was stable at temperature commonly used in invert sugar production. The following thermodynamic parameters were obtained: activation energy (E* d  = 301.57 kJ mol -1 ), enthalpy (298.76 ≤ ΔH* d  ≤ 298.89 kJ mol -1 ), entropy (579.88 ≤ ΔS* d  ≤ 589.27 J K -1 mol -1 ) and Gibbs free energy (100.29 ≤ ΔG* d  ≤ 108.47 kJ mol -1 ). The high E* d , ΔH* d and ΔG* d values confirmed FFase thermostability. The high and positive values for ΔS* d indicate an increase in disorder due opening of the enzyme structure. The sucrose hydrolysis in PBR showed a maximum invert sugar yield (96.0%) at 15 min of operation. The hydrolysis process remained efficient up to 100 min (70.22%). The results obtained in the present study provide a good indication that immobilized FFase on chitosan beads in PBR is efficient to invert sugar production for food industry., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Purpose: To compare the influence of surface roughness and accelerated aging on the mechanical properties of polymer-based and ceramic CAD/CAM materials., Materials and Methods: Three polymers (Lava Ultimate [LVU], VITA ENAMIC [ENA], and Shofu Block HC [SFB]) and one ceramic (IPS Empress CAD [EMP]) were selected for this study. The specimens were treated with the aim of measuring surface roughness (Ra [mm]) and its influence on the mechanical properties. The treatments were: polishing (POL); etching with 9.6% hydrofluoric acid for 90 seconds (ETC); airborne-particle abrasion with 50-μm aluminum oxide at 2-bar pressure (SBT); and airborne-particle abrasion with 50-μm aluminum oxide at 2-bar pressure plus etching with 9.6% hydrofluoric acid (SAC) for 90 seconds. Before and after the accelerated aging (AA) protocol (30,000 cycles, 5°C and 55°C), the specimens were subjected to the 3-point bending test. The data were analyzed with two-way and three-way ANOVA and post hoc Tukey test (P < .05)., Results: The surface roughness ranged from 0.24 μm to 1.96 μm, with statistically significant differences (P < .05). The highest surface roughness, in descending order, was exhibited by: LVU > SFB > EMP > ENA. The AA demonstrated an influence on the flexural strength of LVU, ENA, and SFB. The surface treatment did not affect the flexural moduli of the materials tested. The LVU and SFB showed decreased moduli of resilience after the AA; however, modulus of resilience was not influenced by surface treatment., Conclusion: The polymer-based materials were affected by AA. In addition, surface treatment could jeopardize their mechanical properties in certain conditions.

Pectinex Ultra SP-L, a commercial enzyme preparation with fructosyltransferase activity, was successfully immobilized by covalent binding to Fe 3 O 4 -chitosan- magnetic nanoparticles. Immobilization carried out according to a 2 3 -full factorial design where glutaraldehyde concentration, activation time and time of contact between enzyme and support were selected as the independent variables and immobilization yield as the response. The highest immobilization yield (94.84%) was obtained using 3.0% (v/v) glutaraldehyde and activation and contact times of 180 and 30 min, respectively. The immobilized biocatalyst, which showed for both hydrolytic and transfructosylating activities optimum pH and temperature of 7.0 and 60 °C, respectively, retained 70 and 86% of them after 6 cycles of reuse. A kinetic/thermodynamic study focused on thermal inactivation of the immobilized construct indicated high thermostability at temperatures commonly used for fructo-oligosaccharides (FOS) production. Maximum FOS concentration obtained in lab-scale experiments was 101.56 g L -1 , with predominant presence of 1-kestose in the reaction mixture. The results obtained in this study suggest that the immobilized-enzyme preparation may be effectively exploited for FOS production and easily recovered from the reaction mixture by action of a magnetic field., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Eight different Aspergillus strains were tested for their ability to produce β-fructofuranosidase (FFase) by Solid-State Fermentation. The Aspergillus tamarii URM4634 strain was selected as the most performant and tested on six different agroindustrial by-products. Soy, wheat and oat brans, which allowed for the highest hydrolytic (U H ) and transfructosylating (U TF ) activities, were tested individually or in mixtures according to a simplex-centroid mixture design in order to investigate their effects on FFase production at different times. The best results in terms of both enzyme activities were obtained with only soy bran. The influence of substrate, moisture and sucrose levels on FFase production was evaluated, and the highest U H and U TF activities were 229.43 ± 4.88 and 66.93 ± 3.02 U/mL, respectively. The obtained results indicate that A. tamarii FFase may be a biocatalyst with great potential for industrial applications such as sugar inversion and fructo-oligosaccharides production., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The protease from Aspergillus tamarii Kita UCP1279 extraction by aqueous two-phase PEG-Citrate (ATPS) systems, using a factorial design 2 4 , was investigated. Then, the variables studied were polyethylene glycol (PEG) molar mass (M PEG ), concentrations of PEG (C PEG ) and citrate (C CIT ), and pH. The responses analyzed were the partition coefficient (K), activity yield (Y) and purification factor (PF). The thermodynamic parameters of the ATPS partition were estimated as a function of temperature. ATPS was able to pre-purify the protease (PF = 1.6) and obtained 84% activity yield. The thermodynamic parameters ΔG° m (-10.89 kJ mol -1 ), ΔH m (-5.0 kJ mol -1 ) and partition ΔS m (19.74 J mol -1 K -1 ) showed that the preferential migration of almost all protein contaminants of the crude extract to the salt-rich phase, while the preferred protease was the PEG rich phase. The extracted enzyme presents optimum temperature and pH at range of 40-50 °C and 9.0-11.0, respectively. Moreover, the enzyme was identified as serine protease based on inhibition profile. ATPS showed the satisfactory performance as the first step for Aspergillus tamarii Kita UCP1279 protease pre-purification.

The partitioning and purification of lectins from the crude extract of Cratylia mollis seeds (Cramoll 1,4) was investigated in aqueous two-phase systems (ATPS). A factorial design model (2 4 ) was used to evaluate the influence of polyethylene glycol (PEG) molar mass (1500-8000 g/mol), PEG concentration (12.5-17.5% w/w), phosphate (10-15% w/w) concentration, and pH (6-8) on the differential partitioning, purification factor, and yield of the lectin. Polymer and salt concentration were the most important variables affecting partition of lectin and used to find optimum purification factor by experimental Box-Behnken design together with the response surface methodology (RSM). ATPS showed best conditions composed by 13.9% PEG1500, 15.3% phosphate buffer at pH 6, which ensured purification factor of 4.70. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a single band of protein with 26.1 kDa. Furthermore, results demonstrated a thermostable lectin presenting activity until 60 °C and lost hemagglutinating activity at 80 °C. According to the obtained data it can be inferred that the ATPS optimization using RSM approach can be applied for recovery and purification of lectins.

Aqueous two-phase systems have been studied for almost a century to separate biomolecules in harmless conditions. Proteases produced by Aspergillus tamarii URM 4634 were extracted in polyethylene glycol (PEG)/phosphate aqueous two-phase system under discontinuous and continuous (perforated rotative discs column) process. On the discontinuous process, it was evaluated the effect of operational conditions (PEG molar mass and its concentration, phosphate concentration and pH) over the partition coefficient, activity yield and purification factor. Protease partitioned to PEG-phase with partition coefficients up to 55.73. The best process parameters were 17.5% of PEG, with molar mass 8000 g·mol -1 , 15% of phosphate salt at pH 6, with 113.15% of activity yield and purification factor of 2.62. Under continuous extraction, hold up data showed that 57.1% of the discontinuous phase was available for protein extraction. Further, separation achieved 90.0% of efficiency. The yields surpassed 100% in almost all runs, and the best purification factor was 1.84, with both flows of 2 mL·min -1 . Thus, the best operational conditions reached an activity yield of 95.3% and 90.0% of separation efficiency. Hence, aqueous two-phase system PEG/phosphate extraction is an efficient process for separation of proteases produced by Aspergillus tamarii URM 4634, under continuous extraction likewise under discontinuous process.

Objective: To evaluate the effect of surface treatments on yttria-tetragonal zirconia polycrystal (Y-TZP) characteristics and on resin-mediated zirconia bond., Methods: Y-TZP slabs were grit blasted with 45μm alumina or with 30μm silica-coated alumina particles. The chemical treatments were: no-chemical treatment (NC), silane-containing primer (SP), MDP (10-Methacryloyloxydecyl dihydrogen phosphate) and silane-containing primer (MPS), MDP-containing primer (MP) and MDP and silane-containing adhesive (MPA). Contact angle as a function of surface roughness (θ m ) and surface roughness parameter (Sdr) were measured using Fringe Projection Phase Shifting (FPPS). Surface free energy (γ s TOT ) was calculated with a goniometer. Chemical interaction between primers/adhesive and zirconia was analyzed using time-of-flight secondary ion mass spectrometry (ToF-SIMS). Resin cement microshear bond strength (μSBS) was analyzed at either 24-h or 8-months water storage (37°C). θ m values, Sdr values, γ s TOT and μSBS values were analyzed using Analysis of variance (ANOVA) and post hoc Tukey test (α=0.05)., Results: Chemical treatment had an effect (p<0.001) on all surface parameters analyzed: θ m , γ s TOT and Sdr. MP-treated group showed higher incidence of P-O-Zr bonds than the other groups, indicating more chemical linkages. Grit blasting (p<0.001) and the interaction chemical treatment*storage (p<0.001) did not affect μSBS; all silane-containing primers showed significant drop in μSBS after aging., Significance: MDP and/or silane-based solutions affect the physicochemical properties of blasted-zirconia. An MDP-based primer is fundamental to achieve a stable resin-zirconia bonding, but the chemical reactivity of MDP is impaired when this molecule is present in a multicomponent system., (Copyright © 2019 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.)

This study reports on the biochemical characterization as well as the kinetic and thermodynamic study of Aspergillus tamarii URM4634 β-fructofuranosidase (FFase) with transfructosylating activity. Conditions for FFase activity were optimized by means of a central composite rotational design using pH and temperature as the independent variables, while residual activity tests carried out in the temperature range of 45-65°C enabled us to investigate FFase thermostability and estimate the kinetic and thermodynamic parameters of enzyme denaturation. Optimal conditions for sucrose hydrolysis and fructosyl transfer catalyzed by crude FFase were 50°C, and pH 6.0 and 7.4, respectively. The thermodynamic properties of irreversible enzyme inactivation were found to be activation energy of 293.1 kJ mol -1 , and activation enthalpy, entropy, and Gibbs free energy in the ranges 290.3-290.4 kJ mol -1 , 568.7-571.0 J mol -1 K -1 , and 97.9-108.8 kJ mol -1 , respectively. The results obtained in this study point out satisfactory enzyme activity and thermostability at temperatures commonly used for industrial fructo-oligosaccharide (FOS) synthesis; therefore, this novel FFase appears to be a promising biocatalyst with great potential for long-term FOS synthesis and invert sugar production. To the best of our knowledge, this is the first report on kinetic and thermodynamic parameters of an A. tamarii FFase., (© 2019 American Institute of Chemical Engineers.)

Statement of Problem: Resin-coating agents can be used to fill microstructural defects within composite resin restorations, which might result in a smoother surface. Nevertheless, data about the color stability and surface roughness of coated restorations are lacking., Purpose: The purpose of this in vitro study was to compare the surface roughness and color stability of restorations before and after application of different resin-coating agents., Material and Methods: Forty specimens of composite resin (Filtek Z250) (diameter×height, 6×2 mm) were divided into 5 groups according to the surface treatments (n=8 per group): control (CTR), Fortify (FT), Lasting touch (LT), BisCover (BIS), and Fill Glaze (FG). The specimens were evaluated for surface roughness by using a profilometer with 3 readings for each specimen (separated by 300 μm) before immersion in a staining solution (coffee) and after 5 days immersed in coffee and for color stability (▵E) by using a spectrophotometer before being immersed in coffee (initial reading) and after 6 hours, 12 hours, 1, 2, 3, 4, and 5 days., Results: Two-way ANOVA with repeated measures and the Tukey test showed that the CTR group showed the highest roughness values, similar to those of the FT and LT groups. The BIS and FG groups showed lower roughness than the other groups but similar results when compared with each other. All groups, except LT, showed increased roughness values after immersion in coffee solution, and the BIS group showed the lowest variation over time. Color alterations were found for all groups, with the CTR group showing the lowest ▵E values when compared with the other groups except for the BIS group. No correlation was found between surface roughness and degree of color change (P=.401)., Conclusions: The BisCover surface sealant showed the lowest surface roughness and the best color stability. Surface sealants can be used to fill microcracks and microgaps but should not be used as a substitute for polishing., (Copyright © 2019 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Purpose: To evaluate the effect of endodontic retreatment on the bond strength of different cementation techniques (self-adhesive and conventional resin cement) through the push-out mechanical testing and the penetrability of resin cements to root dentin., Methods: 60 human teeth with single oval-shaped canals were used (n=15) : G1 - Endodontic treatment and cementation with RelyX ARC (ETA); G2 - Endodontic treatment and cementation with U200 (ETU); G3 - Endodontic retreatment and cementation with RelyX ARC (ERA); G4 - Endodontic retreatment and cementation with U200 (ERU). The groups with conventional endodontic treatment were filled with AH plus (ETA and ETU), while the groups that were submitted to endodontic retreatment were initially filled with Endofill and afterwards with AH Plus. Each radicular third (cervical, middle, and apical, of each tooth) was submitted to push-out bond strength testing, followed by evaluation with confocal laser microscopy to determine the penetration of the resin cements, and scanning electron microscopy was used to evaluate the failure mode. The parametric data were evaluated by two-way ANOVA and Tukey tests, and non-parametric data by Kruskal-Wallis and Dunn tests (α< 0.05)., Results: The ETA group had higher bond strength values to intra-radicular dentin than the other groups in all root thirds (P< 0.05). Endodontic retreatment had a negative impact on the penetration and push-out bond strength of RelyX ARC and U200 cement, regardless of the analyzed radicular third., Clinical Significance: The bond strength between fiber post and root dentin can be affected by several factors, such as technical failure, cementation technique, pretreatment of the dentin, type of post and adaptation. The endodontic retreatment interferes negatively on the bond strength and penetrability of RelyX ARC and U200 cements to dentin, regardless of the analyzed radicular third. Therefore, the endodontic retreatment might have an adverse effect due to over preparation and aggression to the root canal., Competing Interests: The authors declared no conflict of interest., (Copyright©American Journal of Dentistry.)

The kinetics and thermodynamics of pectin hydrolysis in cashew apple juice by polygalacturonase (PG) from Aspergillus aculeatus URM4953 covalently-immobilized on calcium alginate beads were investigated. Immobilized-PG activity in cashew apple juice was the highest at 20 °C, showing a maximum hydrolysis rate of 58.2 mg/mL·min, a catalytic constant of 166.2 s -1 and an affinity constant of 113.0 mg/mL. Since the enzyme exhibited an allosteric behavior, the hydrolysis rate was modeled, with excellent accuracy, by the Hill Equation as function of pectin concentration. The Hill coefficient increased from 3 to 5 with increasing temperature from 20 to 50 °C, evidencing a positive cooperativity mechanism. The reaction activation energy and the standard enthalpy variation of enzyme unfolding were 80.3 and 16.6 kJ/mol, respectively. Consistently with the kinetic results, PG-catalyzed pectin hydrolysis proceeded with maximum spontaneity at 20 °C, showing activation Gibbs free energy, enthalpy and entropy of 59.3 kJ/mol, 77.9 kJ/mol and 63.4 J/mol·K, respectively. Immobilized PG was successful in the hydrolysis of cashew apple juice pectin, requiring a low temperature to act optimally., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Computer-aided design/computer-aided manufacturing (CAD/CAM) materials were submitted to thermocycling, to identify changes in mechanical behavior. Four CAD/CAM materials were divided in four subordinate groups (n=9): (1) dry out for 7 days, (2) distilled water at 37 o C for 7 days, (3) 60,000 thermocycles, and (4) 120,000 thermocycles. Following thermocycling, samples were submitted to three-point bending test. Two-way ANOVA and post-hoc Tukey's test were performed (α=0.05). The IPS e.max CAD had a flexural strength of 396±75 MPa and flexural modulus of 84±11 GPa, followed by Vita Enamic with values of 153±17 MPa and 28±5 GPa respectively. The flexural strength recorded for Lava Ultimate was 149±28 MPa and the flexural modulus was 12±3 GPa. Vitablocs Mark II had the lowest flexural strength values (125±10 MPa) and a flexural modulus of 49±15 GPa. Although polymer-based materials have similar mechanical properties compared to ceramics, they are affected by thermo cycling conditions.

The aim of this study is to evaluate the machinability of four CAD/CAM materials (n = 13) assessed by brittleness index, Vickers hardness, and fracture toughness and interaction among such mechanical properties. The materials selected in this in vitro study are Feldspathic ceramic [FC], Lithium-disilicate glass ceramic [LD], leucite-reinforced glass ceramic [LR], and nanofilled resin material [RN]. Slices were made from the blocks following original dimensions 14 × 12 × 3 mm (L × W × H), using a precision slow-speed saw device and then surfaces were regularized through a polishing device. Brittleness index and fracture toughness were calculated by the use of specific equations for each one of the properties. The Vickers hardness was calculated automated software in the microhardness device. One-way Anova and Pearson's correlation were applied to data evaluation. LD obtained the highest values for brittleness index and was not significantly different from FC. LR presented statistically significant difference compared with RN, which had the lowest mean. Vickers hardness showed LD with the highest average, and no statistical difference was found between FC and LR. RN presented the lowest average. Fracture toughness showed FC and LR not statistically different from each other, likewise LD and RN. The brittleness index, considered also as the machinability of a material, showed within this study as positively dependent on Vickers hardness, which leads to conclusion that hardness of ceramics is related to its milling capacity. In addition, fracture toughness of pre-sintered ceramics is compared to polymer-based materials.

Circular dichroism (CD) and fluorescence spectroscopy (FS) were used to monitor the pH-dependent conformational and structural stability changes induced by temperature and UV light on the protease from Aspergillus tamarii URM4634 at different pH values. The formation of photoproducts, such as N-formylkynurenine, dityrosine and kynurenine, were monitored with FS. The pH-dependent melting temperatures (T m ) were determined using CD and FS from 20 to 90 °C. Conformational changes were correlated with the pH-dependent biochemical activities. CD revealed that the protease is rich in α-helices. Thermal denaturation was irreversible at all pH range and displayed T m values from 42.8 to 67.8 °C (CD) and from 38 to 60.3 °C (FS), which the highest T m was observed at pH 6. The light and temperature induced to the formation of photoproducts was more intense at high pH value. Despite the biochemical data shows optimum pH 9, the highest stability was at pH 6, maintaining 100% of activity after 24 h. The acquired data permits to select the best physicochemical parameters to secure the optimal activity and stability when used in biotechnological applications. Furthermore, the conformal changes induced by temperature in the protein are directly correlated with its level of biochemical activity., (Copyright © 2018 Elsevier B.V. All rights reserved.)

An extracellular serine-protease from Aspergillus tamarii URM4634 was purified and characterized. The possibility of using Aspergillus tamarii URM4634 protease in detergent formulations and collagenolytic activity was investigated. The protease demonstrated excellent stability at pH range 7.0-11.0, the optimum being at pH 9.0. The enzyme was stable at 40 °C for 180 min, enhanced by Mg ++ and Ca ++ , but inhibited by Zn ++ , and strongly inhibited by phenylmethylsulfonyl fluoride (PMSF), suggested as serine-protease. The azocasein substrate result showed Km = 0.434 mg/mL and V max  = 7.739 mg/mL/min. SDS-PAGE and azocasein zymography showed that the purified alkaline protease (2983.8 U/mg) had a molecular mass of 49.3 kDa. The enzyme was purified by column chromatography using Sephadex A50 resin. The proteolytic activity was activated by SDS (sodium dodecyl sulfate), Tween-80, Tween 20 and Triton-100. This study demonstrated that A. tamarii URM4634 protease has potent, stable and compatible collagenolytic activity to the desired level in local laundry detergent brands compared with similar enzymes produced by solid-state fermentation. This protease can thus be chosen as an option in both the food industry to tenderization meat and the detergent industry to washing process., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The purpose of this in vitro study was to compare the micro hardness, color stability/ΔE, and degree of conversion/DC of a resin cement light cured through different ceramic spacers. Lithium-disilicate ceramic samples were obtained from IPS E-max CAD blocks (HT A1) and IPS in-Ceram (transparent neutral); and divided in 7 groups (n = 8 for each test): CTR/control group; 06 M/0.6 mm monolithic; 12 M/1.2 mm monolithic; 20 M/2.0 mm monolithic; 06B/0.4 + 0.2 mm bilayered; 12B/1.0 + 0.2 mm bilayered; 20B/1.8 + 0.2 mm bilayered. The resin cement (Variolink veneer) was light cured through the ceramic spacers. The resin cement samples were evaluated for ΔE using a spectrophotometer after 24 h, 7days and after aging (24 h in water at 60 °C). Knoop microhardness and DC tests were conducted immediately after light curing, after 24 h and 7days. All experimental groups showed similar microhardness values, although being lower than CTR group. Similar results were observed after 7days. ΔE was similar between all groups after 24 h (except for 12B and 20B), and increased for all groups after 7days and after artificial aging, especially for thicker and bilayer groups. Only 06 M showed values similar to CTR group. DC values were similar to all groups immediately after light curing, increasing after 24 h and 7days. After 7days, only group 20B showed lower DC than CTR group. A tendency of higher DC could be observed for monolithic and thinner ceramics. All test results showed strong correlation (0.9987). Ceramic interposition can reduce mechanical and physical properties of resin cements, especially with thicker and bilayered ceramics. Group 06 M showed the best ΔE overtime., (Published by Elsevier Ltd.)

The kinetics and thermodynamics of Aspergillus aculeatus pectinase, either free or immobilized in alginate beads, were investigated. Pectinase immobilization ensured an enzyme immobilization yield of 59.71%. The irreversible denaturation of pectinase in both preparations was evaluated at temperatures ranging from 30 to 60 °C. When temperature was raised, the first-order thermal denaturation constant increased from 0.0011 to 0.0231 min -1 for the free enzyme and from 0.0017 to 0.0700 min -1 for the immobilized one, respectively. The results of residual activity tests enabled us to estimate, for denaturation of both free and immobilized pectinase, the activation energy (E ⁎ d  = 85.1 and 101.6 kJ·mol -1 ), enthalpy (82.59 ≤ ΔH ⁎ d  ≤ 82.34 kJ·mol -1 and 99.11 ≤ ΔH ⁎ d  ≤ 98.86 kJ·mol -1 ), entropy (-63.26 ≤ ΔS ⁎ d  ≤ -63.85 J·mol -1 ·K -1 and -5.50 ≤ ΔS ⁎ d  ≤ -5.23 J·mol -1 ·K -1 ) and Gibbs free energy (101.8 ≤ ΔG ⁎ d  ≤ 104.7 kJ·mol -1 and 100.6 ≤ ΔG ⁎ d  ≤ 102.0 kJ·mol -1 ). The integral activity of a continuous system using the free and immobilized enzyme was also predicted, whose results indicated a satisfactory enzyme long-term thermostability in both preparations at temperatures commonly used to clarify juice. These results suggest that both free and immobilized pectinase from A. aculeatus may be profitably exploited in future food industrial applications, with special concern to the immobilized enzyme because of its reusability., (Copyright © 2018 Elsevier B.V. All rights reserved.)