Evaluation of shade correspondence between high-translucency pre-colored zirconia and shade tab by considering the influence of cement shade and substrate materials.

Statement of problem: The relation between cement color and abutment substrate material and the corresponding effect on the color accuracy of high-transparency pre-colored zirconia (HT-Zr) remains unclear. Purpose: This in-vitro study aimed to investigate the difference in color accuracy when the HT-Zr is bonded to different materials-based substrates with differently colored resin cement. Materials and methods: Vita A1 shade HT-Zr with 1 mm thickness was used as the testing sample. The samples were first placed on zirconia (ZR), tooth color resin (CR), and metallic (MT) abutment substrates. Subsequently, four differently colored cements (translucent (TR), bleach, opaque, and A2 shade (A2)) were used for bonding HT-Zr onto the substrate, and the non-bonded group was used as the control group (CG). There were 15 groups in total (n = 10 per group). A digital colorimeter was used to obtain Commission Internationale de l'Eclairage (CIELab) color parameters. The translucency parameter (TP00) of the substrate and sample, as well as color difference (ΔE00) and chroma (C) between the different groups were calculated. Additionally, the ΔE00 and TP00 were compared with the moderately unacceptable match of ΔE00 = 3.6. The statistical analysis was conducted using ANOVA and Tukey HSD post-hoc test (α = 0.05). Results: HT-Zr exhibited high translucency (TP00 = 11.02 ± 0.18), and the mean ΔE00 of the testing samples ranged between 2.18 ± 0.20 and 13.14 ± 0.31. The ZR-CG and MT-A2 groups showed the highest and lowest lightness separately. The CR-CG group exhibited the highest C, and the ΔE00 was lower than that of 3.6. The MT-TR group showed the lowest C and the highest ΔE00. The inter-group comparison revealed that the ΔE00 for different cement is mostly lower than the acceptable color match of 1.0; moreover, the ΔE00 for all the substrates, excluding the CG group, is higher than 3.6. Conclusions: The abutment substrate materials and the cement color should be considered with caution when using HT-Zr, with the effect of abutment substrate materials being more apparent in color accuracy. HT-Zr restorations are not recommended for discolored or bleached abutments but only for natural-colored abutments to achieve the optimal color appearance.

Enhancing Dental Cement Bond Strength with Autofocus-Laser-Cutter-Generated Grooves on Polyetheretherketone Surfaces.

Polyetheretherketone (PEEK) is widely used in dentistry owing to its exceptional properties, including its natural appearance; however, existing surface treatment methods for bonding PEEK have limitations. Autofocus laser cutters, known for their precise engraving and cutting capabilities, offer potential for surface treatment of PEEK; thus, the objective of this study was to investigate the creation of laser groove structures on PEEK to enhance its bonding capability with dental resin cement. A dental computer-aided design and manufacturing system was used to fabricate PEEK samples, and three groove patterns (circle, line, and grid) were generated on PEEK surfaces, with air-abrasion used as the control group. The surface characteristics, cell viability, and bond strength were evaluated, and the data were statistically analyzed using one-way analysis of variance and post hoc Tukey's tests (α = 0.05). Laser-treated PEEK exhibited a uniform texture with a groove depth of approximately 39.4 µm, hydrophobic properties with a contact angle exceeding 90°, a surface roughness of 7.3-12.4 µm, consistent topography, and comparable cell viability compared with untreated PEEK. Despite a decrease in bond strength after thermal cycling, no significant intergroup differences were observed, except for the line-shaped laser pattern. These findings indicate that the autofocus laser cutter effectively enhances the surface characteristics of PEEK by creating a uniform texture and grooves, showing promise in improving bonding properties, even considering the impact of thermal cycling effects.

Development and Assessment of a Novel Whole-Gene-Based Targeted Next-Generation Sequencing Assay for Detecting the Susceptibility of Mycobacterium tuberculosis to 14 Drugs.

Targeted next-generation sequencing (tNGS) has emerged as an alternative method for detecting drug-resistant tuberculosis (DR-TB). To provide comprehensive drug susceptibility information and to address mutations missed by available commercial molecular diagnostics, we developed and evaluated a tNGS panel with 22 whole-gene targets using the Ion Torrent platform to predict drug resistance to 14 drugs, namely, rifampicin (RIF), isoniazid (INH), ethambutol (EMB), pyrazinamide (PZA), moxifloxacin (MFX), levofloxacin (LFX), amikacin (AMK), capreomycin (CM), kanamycin (KM), streptomycin (SM), bedaquiline (BDQ), clofazimine (CFZ), linezolid (LZD), and delamanid (DLM). We selected 50 and 35 Mycobacterium tuberculosis isolates with various DR profiles as the training set and the challenge set, respectively. Comparative variant analyses of the DR genes were performed using Sanger sequencing and whole-genome sequencing (WGS). Phenotypic drug susceptibility testing (pDST) results were used as gold standards. Regarding the limit of detection, the tNGS assay detected 2.9 to 3.8% minority variants in 4% mutant mixtures. The sensitivity and specificity of tNGS were 97.0% (95% confidence interval [CI] = 93.1 to 98.7%) and 99.1% (95% CI = 97.7 to 99.7%), respectively. The concordance of tNGS with pDST was 98.5% (95% CI = 97.2 to 99.2%), which was comparable to that of WGS (98.7%, 95% CI = 97.4 to 99.3%) and better than that of Sanger sequencing (96.9%, 95% CI = 95.3 to 98.0%). The agreement between tNGS and pDST was almost perfect for RIF, INH, EMB, MFX, LFX, AMK, CM, KM, SM, BDQ, and LZD (kappa value = 0.807 to 1.000) and substantial for PZA (kappa value = 0.791). Our customized novel whole-gene-based tNGS panel is highly consistent with pDST and WGS for comprehensive and accurate prediction of drug resistance in a strengthened and streamlined DR-TB laboratory program. IMPORTANCE We developed and validated a tNGS assay that was the first to target 22 whole genes instead of regions of drug resistance genes and comprehensively detected susceptibility to 14 anti-TB drugs, with great flexibility to include new or repurposed drugs. Notably, we demonstrated that our custom-designed Ion AmpliSeq TB research panel platform had high concordance with pDST and could significantly reduce turnaround time (by approximately 70%) to meet a clinically actionable time frame. Our tNGS assay is a promising DST solution for providing needed clinical information for precision medicine-guided therapies for DR-TB and allows the rollout of active pharmacovigilance.

Primary Bedaquiline Resistance Among Cases of Drug-Resistant Tuberculosis in Taiwan.

Bedaquiline (BDQ), which is recommended for the treatment of drug-resistant tuberculosis (DR-TB), was introduced in Taiwan in 2014. Due to the alarming emergence of BDQ resistance, we conducted BDQ resistance analyses to strengthen our DR-TB management program. This retrospective population-based study included initial Mycobacterium tuberculosis isolates from 898 rifampicin-resistant (RR) or multidrug-resistant (MDR) TB cases never exposed to BDQ during 2008-2019. We randomly selected 65 isolates and identified 28 isolates with BDQ MIC<0.25µg/ml and MIC≥0.25µg/ml as the control and study groups, respectively. BDQ drug susceptibility testing (DST) using the MGIT960 system and Sanger sequencing of the atpE, Rv0678, and pepQ genes was conducted. Notably, 18 isolates with BDQ MIC=0.25µg/ml, 38.9% (7/18), and 61.1% (11/18) isolates were MGIT-BDQ resistant and susceptible, respectively. Consequently, we recommended redefining MIC=0.25µg/ml as an intermediate-susceptible category to resolve discordance between different DST methods. Of the 93 isolates, 22 isolates were MGIT-BDQ-resistant and 77.3% (17/22) of MGIT-BDQ-resistant isolates harbored Rv0678 mutations. After excluding 2 MGIT-BDQ-resistant isolates with borderline resistance (GU400growth control-GU100BDQ≤1day), 100% (15/15) harbored Rv0678 gene mutations, including seven novel mutations [g-14a, Ile80Ser (N=2), Phe100Tyr, Ala102Val, Ins g 181-182 frameshift mutation (N=2), Del 11-63 frameshift mutation, and whole gene deletion (N=2)]. Since the other 22.7% (5/22) MGIT-BDQ-resistant isolates with borderline resistance (GU400growth control-GU100BDQ≤1day) had no mutation in three analyzed genes. For isolates with phenotypic MGIT-BDQ borderline resistance, checking for GU differences or conducting genotypic analyses are suggested for ruling out BDQ resistance. In addition, we observed favorable outcomes among patients with BDQ-resistant isolates who received BDQ-containing regimens regardless of Rv0678 mutations. We concluded that based on MIC≥0.25µg/ml, 3.1% (28/898) of drug-resistant TB cases without BDQ exposure showed BDQ resistance, Rv0678 was not a robust marker of BDQ resistance, and its mutations were not associated with treatment outcomes.

Dual expression of transgenic delta-5 and delta-6 desaturase in tilapia alters gut microbiota and enhances resistance to Vibrio vulnificus infection.

Omega-3 polyunsaturated fatty acids (n-3 PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), exhibit antibacterial and anti-inflammatory activities. Furthermore, diets rich in n-3 PUFAs are known to improve disease resistance and limit pathogen infection in commercial aquaculture fishes. In this study, we examined the effects of transgenic overexpression of n-3 PUFA biosynthesis genes on the physiological response to bacterial infection in tilapia. We first established tilapia strains with single or dual expression of salmon delta-5 desaturase and/or delta-6 desaturase and then challenged the fish with Vibrio vulnificus infection. Interestingly, our data suggest that n-3 PUFA-mediated alterations in gut microbiota may be important in determining disease outcome via effects on immune response of the host. Both liver- and muscle-specific single and dual expression of delta-5 desaturase and delta-6 desaturase resulted in higher n-3 PUFA content in transgenic fish fed with a LO basal diet. The enrichment of n-3 PUFAs in dual-transgenic fish is likely responsible for their improved survival rate and comparatively reduced expression of inflammation- and immune-associated genes after V. vulnificus infection. Gut microbiome analysis further revealed that dual-transgenic tilapia had high gut microbiota diversity, with low levels of inflammation-associated microbiota (i.e., Prevotellaceae). Thus, our findings indicate that dual expression of transgenic delta-5 and delta-6 desaturase in tilapia enhances disease resistance, an effect that is associated with increased levels of n-3 PUFAs and altered gut microbiota composition.

Granulin peptide GRN-41 of Mozambique tilapia is a novel antimicrobial peptide against Vibrio species.

In our previous study, the novel GRN-41 peptide generated from alternative splicing of the Mozambique tilapia PGRN1 gene was identified to be a potent peptide that protected against V. vulnificus in the transgenic zebrafish model by modulating innate immune-related genes. In this study, the anti-bacterial activities of synthetic Mozambique tilapia GRN-41 peptide (OmGRN-41) against various bacterial pathogens were investigated. The results showed that OmGRN-41 had bactericidal activity against Vibrio species, including V. vulnificus, V. alginolyticus, and V. harveyi, but exhibited bacteriostatic activity against V. parahaemolyticus. OmGRN-41 maintained bactericidal activity (64 µM) against V. vulnificus at pH 2 to pH 10 or after heat treatment for 1 h at high temperatures between 40 °C and 100 °C. TEM observations revealed that the outer membrane of V. vulnificus was disrupted by OmGRN-41, leading to morphological rupture and loss of cytoplasmic contents. Additionally, little hemolytic activity against tilapia and sheep erythrocytes was detected after treatment with 128 µM OmGRN-41. OmGRN-41 can effectively enhance the survival of Nile tilapia infected by V. vulnificus. Our results suggest that the OmGRN-41 is a novel antimicrobial peptide possessing bactericidal activity, especially against Vibrio species. These results indicate that OmGRN-41 can be applied in human Vibriosis treatment and has the potential to defend against Vibrio spp. infection in critical aquaculture organisms.

A potent tilapia secreted granulin peptide enhances the survival of transgenic zebrafish infected by Vibrio vulnificus via modulation of innate immunity.

Progranulin (PGRN) is a multi-functional growth factor that mediates cell proliferation, survival, migration, tumorigenesis, wound healing, development, and anti-inflammation activity. A novel alternatively spliced transcript from the short-form PGRN1 gene encoding a novel, secreted GRN peptide composed of 20-a.a. signal peptide and 41-a.a. GRN named GRN-41 was identified to be abundantly expressed in immune-related organs including spleen, head kidney, and intestine of Mozambique tilapia. The expression of GRN-41 and PGRN1 were further induced in the spleen of tilapia challenged with Vibrio vulnificus at 3 h post infection (hpi) and 6 hpi, respectively. In this study, we established three transgenic zebrafish lines expressing the secreted GRN-41, GRN-A and PGRN1 of Mozambique tilapia specifically in muscle. The relative percent of survival (RPS) was enhanced in adult transgenic zebrafish expressing tilapia GRN-41 (68%), GRN-A (32%) and PGRN1 (36%) compared with control transgenic zebrafish expressing AcGFP after challenge with V. vulnificus. It indicates tilapia GRN-41 is a potent peptide against V. vulnificus infection. The secreted tilapia GRN-41 can induce the expression of innate immune response-related genes, such as TNFa, TNFb, IL-8, IL-1ß, IL-6, IL-26, IL-21, IL-10, complement C3, lysozyme (Lyz) and the hepatic antimicrobial peptide hepcidin (HAMP), in adult transgenic zebrafish without V. vulnificus infection. The tilapia GRN-41 peptide can enhance the innate immune response by further elevating TNFb, IL-1ß, IL-8, IL-6, and HAMP expression in early responsive time to the V. vulnificus challenge in transgenic zebrafish. Our results suggest that the novel GRN-41 peptide generated from alternative splicing of the tilapia PGRN1 gene is a potent peptide that defends against V. vulnificus in the transgenic zebrafish model by modulation of innate immunity.

Complete Genome Sequence of Streptococcus iniae 89353, a Virulent Strain Isolated from Diseased Tilapia in Taiwan.

Streptococcus iniae 89353 is a virulent strain isolated from diseased tilapia in Taiwan. The full-genome sequence of S. iniae 89353 is 2,098,647 bp. The revealed genome information will be beneficial for identification and understanding of potential virulence genes of Streptococcus iniae and possible immunogens for vaccine development against streptococcosis.