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Objectives: Traditional fixed prosthodontic training evaluations are time-consuming and subjective. This study aimed to introduce and evaluate a novel 3D auto-evaluate tooth preparation with an augmented reality (3DAR) visualization algorithm to enhance dental education for students and improve the efficiency of fixed prosthodontic training evaluations., Methods: Fifty maxillary central incisors and first molars prepared by 50 dental students on typodont models were scanned with a 3D scanner to capture STL files and evaluated using the 3DAR algorithm, which calculated Euclidean distances and root mean square error (RMSE) for accuracy assessment and assigned scores based on RMSE, using an augmented reality (AR) app for interactive evaluation and visualization. These scores were compared to manual scoring with computer assistance (MSCA method), which also used RMSE but required manual alignment and model scoring. Intrarater and interrater reliability of the 3DAR scoring method was assessed using intraclass correlation coefficients (ICC) and compared with the MSCA method. Additionally, the feasibility of 3DAR was evaluated based on evaluation time and user satisfaction., Results: The 3DAR method demonstrated good-to-excellent interrater agreement (ICC = 0.75-0.95) and perfect intrarater reliability (ICC = 1), while the MSCA method showed moderate-to-good reliability (ICC = 0.74-0.89). 3DAR significantly reduced evaluation time (10.514 s vs. 2 h required for MSCA) and received high user satisfaction ratings (average score = 4.66±0.24)., Conclusions: The 3DAR algorithm offers a reliable and efficient assessment method for prosthodontic training. It showed strong agreement with traditional methods, significantly reduced evaluation time, and achieved high user satisfaction., Clinical Significance: The integration of a 3D augmented reality-based auto-evaluation algorithm into tooth preparation training enhances reliability and provides interactive, real-time feedback on performance, highlighting its potential to advance dental education practices., 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 Ltd. All rights reserved.)

Hepatopancreatic microsporidiosis (HPM), caused by the microsporidium Ecytonucleospora hepatopenaei (EHP) leads to retarded growth and enhanced susceptibility to other diseases in shrimp resulting in a major loss for the shrimp industry worldwide. It is little understood how EHP infects its host and hijacks its cellular machinery to replicate and exert clinical manifestations in infected shrimp. Since the initial record of HPM, histopathology and polymerase chain reaction (PCR)-based assays were developed for the detection of EHP to prevent spread of the disease. Availability of an antibody-based detection method would complement these existing diagnostic tools and be useful in studying EHP pathogenesis. We describe here an immunofluorescence assay (IFA) for detecting EHP using monoclonal antibodies (mAbs) that were originally developed against Cryptosporidium parvum, a coccidian parasite that infects calves (Bos taurus), other agriculturally important animals, and humans. Forty-one mAbs were screened and two mAbs, 3E2 and 3A12, were found to detect EHP successfully. The utility of these mAbs in detecting EHP was further assessed by testing 36 experimentally challenged EHP-infected shrimp (Penaeus vannamei). EHP-detection data from infected shrimp were compared by Hematoxylin and Eosin (H&E) histology, real-time PCR, and immunofluorescence. The data show IFA using mAbs 3E2 and 3A12 could successfully detect EHP and that the sensitivity of detection is comparable to H&E histology and quantitative PCR. Availability of mAbs that can detect EHP is expected to be immensely beneficial in HPM diagnosis. Since the pathobiology of C. parvum has been so widely studied, these cross-reactive mAbs may also aid in gaining some insight into EHP pathogenesis and disease., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could appear to influence the work reported in this study., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Neurodegenerative diseases are central or peripheral nervous system disorders associated with progressive brain cell degeneration. Common neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis have been widely studied. However, current therapeutics only reduce the symptoms and do not ameliorate the pathogenesis of these diseases. Recent studies suggested the roles of neuroinflammation, apoptosis, and oxidative stress in neurodegenerative diseases. Mesenchymal stem cells (MSCs) exert anti-apoptotic, anti-inflammatory, and antioxidative effects. Therefore, investigating the effects of MSCs and their applications may lead to the discovery of more effective therapies for neurodegenerative diseases. In this study, we review different approaches used to identify therapies for neurodegenerative diseases using MSCs., Competing Interests: 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., (© 2024 The Author(s).)

This study aims to genomically elucidate six isolates of rapidly growing non-tuberculous mycobacteria (RGM) derived from Siamese fighting fish ( Betta splendens ). These isolates had previously undergone phenotypic and biochemical characterization, antibiotic susceptibility testing, and in vivo virulence assessment. Initial DNA barcoding using the 16S rRNA sequence assigned these six isolates to five different species, namely Mycobacterium chelonae (BN1983), M. cosmeticum (BN1984 and N041), M. farcinogenes (SNSK5), M. mucogenicum (BN1956), and M. senegalense (BN1985). However, the identification relied solely on the highest percent identity of the 16S rRNA gene, raising concerns about the taxonomic ambiguity of these species. Comprehensive whole genome sequencing (WGS) and extended genomic comparisons using multilocus sequence typing (MLST), average nucleotide identity (ANI), and digital DNA-DNA hybridization (dDDH) led to the reclassification of BN1985 and SNSK5 as M. conceptionense while confirming BN1983 as M. chelonae and BN1984 and N041 as M. cosmeticum . Notably, the analysis of the BN1956 isolate revealed a potential new species that is proposed here as M. mucogenicum subsp. phocaicum sp. nov. Common genes encoding "mycobacterial" virulence proteins, such as PE and PPE family proteins, MCE, and YrbE proteins, were detected in all six isolates. Two species, namely M. chelonae and M. cosmeticum , appear to have horizontally acquired T6SS-II ( clpB ), catalase ( katA ), GroEL ( groel ), and capsule ( rmlb ) from distantly related environmental bacteria such as Klebsiella sp., Neisseria sp., Clostridium sp., and Streptococcus sp. This study provides the first draft genome sequence of RGM isolates currently circulating in B. splendens and underscores the necessity of WGS for the identification and classification of mycobacterial species.

Purpose: To evaluate the reliability of various reference areas for digital alignment between 3D resting and smiling facial models., Materials and Methods: 3D posed and natural smiling faces of 33 adults were registered to the respective neutral faces, using six matching strategies with different reference matching surfaces: nose (N), nose + central forehead (NFc), nose + whole forehead (NFw), nose + chin (NC), nose + central forehead + chin (NFcC), and nose + whole forehead + chin (NFwC). The positional discrepancies of the registered images were measured at the left and right pupil centers., Results: Two-way ANOVA and post hoc multiple pairwise t test with Bonferroni correction (α = .05) were used to evaluate the measurements. As a result, the use of larger reference areas increases the trueness of image-matching, whereas there was no statistically significant difference between the matching strategies within the same smiling type. Meanwhile, the image registration of posed smiles resulted in fewer positional disparities than the natural smiles with significant differences observed for the registration using the NC and NFcC surface-based matching areas at the right pupil (P = .030 and .026, respectively)., Conclusions: The findings of this study suggest that the reference surface areas and smiling types have some impact on the accuracy of 3D smiling facial image alignments. Large and evenly distributed matching surfaces are recommended for posed smiles, whereas caution should be taken when using the chin area as a reference surface for matching natural smile facial images.

Aim: The current study aimed to compare the responses and satisfaction reported by users with varying levels of experience when using different types of computer-aided design (CAD) software programs to design crowns., Materials and Methods: A questionnaire was used to evaluate user responses to five domains (software visibility, 3Dscanned data preparation, crown design and adjustment, finish line registration, and overall experience) of various CAD software programs. The study included 50 undergraduate dental students (inexperienced group) and 50 dentists or dental technicians from two hospitals (experienced group). The participants used four different CAD software programs (Meshmixer, Exocad, BlueSkyPlan, and Dentbird) to design crowns and recorded the features using the questionnaire. Statistical analyses included one-way and two-way analysis of variance (ANOVA) tests to compare scores and verify the interaction between user response and experience., Result: User evaluation scores in the domains of software visibility and 3D-scanned data preparation varied between software programs (P < 0.001), with Exocad being favored by the experienced group. When evaluating crown design and finish line registration, Dentbird and Exocad scored significantly higher than the other software in both groups as they offered automation of the process using deep learning (P < 0.001). Two-way ANOVA showed that prior experience of using CAD significantly affected the users' responses to all queries (P < 0.001)., Conclusion: User response and satisfaction varied with the type of CAD software used to design dental prostheses, with prior experience of using CAD playing a significant role. Automation of design functions can enhance user satisfaction with the software.

Purpose: This study aimed to examine the satisfaction of dental professionals, including dental students, dentists, and dental technicians, with computer-aided design (CAD) software performance using deep learning (DL) and explainable artificial intelligence (XAI)-based behavioral analysis concepts., Materials and Methods: This study involved 436 dental professionals with diverse CAD experiences to assess their satisfaction with various dental CAD software programs. Through exploratory factor analysis, latent factors affecting user satisfaction were extracted from the observed variables. A multilayer perceptron artificial neural network (MLP-ANN) model was developed along with permutation feature importance analysis (PFIA) and the Shapley additive explanation (Shapley) method to gain XAI-based insights into individual factors' significance and contributions., Results: The MLP-ANN model outperformed a standard logistic linear regression model, demonstrating high accuracy (95%), precision (84%), and recall rates (84%) in capturing complex psychological problems related to human attitudes. PFIA revealed that design adjustability was the most important factor impacting dental CAD software users' satisfaction. XAI analysis highlighted the positive impacts of features supporting the finish line and crown design, while the number of design steps and installation time had negative impacts. Notably, finish-line design-related features and the number of design steps emerged as the most significant factors., Conclusions: This study sheds light on the factors influencing dental professionals' decisions in using and selecting CAD software. This approach can serve as a proof-of-concept for applying DL-XAI-based behavioral analysis in dentistry and medicine, facilitating informed software selection and development., (© 2024 by the American College of Prosthodontists.)

Statement of Problem: Tooth preparation is an essential part of prosthetic dentistry; however, traditional evaluation methods involve subjective visual inspection that is prone to examiner variability., Purpose: The purpose of this study was to investigate a newly developed automated scoring and augmented reality (ASAR) visualization software program for evaluating tooth preparations., Material and Methods: A total of 122 tooth models (61 anterior and 61 posterior teeth) prepared by dental students were evaluated by using visual assessments that were conducted by students and an expert, and auto assessment that was performed with an ASAR software program by using a 3-dimensional (3D) point-cloud comparison method. The software program offered comprehensive functions, including generating detailed reports for individual test models, producing a simultaneous summary score report for all tested models, creating 3D color-coded deviation maps, and forming augmented reality quick-response (AR-QR) codes for online data storage with AR visualization. The reliability and efficiency of the evaluation methods were measured by comparing tooth preparation assessment scores and evaluation time. The data underwent statistical analysis using the Kruskal-Wallis test, followed by Mann-Whitney U tests for pairwise comparisons adjusted with the Benjamini-Hochberg method (α=.05)., Results: Significant differences were found across the evaluation methods and tooth types in terms of preparation scores and evaluation time (P<.001). A significant difference was observed between the auto- and student self-assessment methods (P<.001) in scoring both the anterior and posterior tooth preparations. However, no significant difference was found between the auto- and expert-assessment methods for the anterior (P=.085) or posterior (P=.14) tooth preparation scores. Notably, the auto-assessment method required significantly shorter time than the expert- and self-assessment methods (P<.001) for both tooth types. Additionally, significant differences in evaluation time between the anterior and posterior tooth were observed in both self- and expert-assessment methods (P<.001), whereas the evaluation times for both the tooth types with the auto-assessment method were statistically similar (P=.32)., Conclusions: ASAR-based evaluation is comparable with expert-assessment while exhibiting significantly higher time efficiency. Moreover, AR-QR codes enhance learning and training experiences by facilitating online data storage and AR visualization., (Copyright © 2024 Editorial Council for The Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Introduction: Virtual reality-based interactive simulation (VRIS) provides a safe and controlled environment for dental students and professionals to develop skills and knowledge. This study aimed to investigate the effectiveness of using the VRIS for prosthodontic practice and to explore the trends, application areas, and users' attitudes towards VRIS., Materials and Methods: This review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines for searching studies published until 21 March 2023 that reported quantitative or qualitative learning outcomes related to the use of VRIS for dental prosthodontic practice and clinical training. The quality of the included studies was assessed using the Medical Education Research Study Quality Instrument (MERSQI) and Newcastle-Ottawa Scale-Education (NOS-E) tools. A random-effects meta-analysis was conducted to compare the intervention group (utilizing VRIS) and the control group (employing conventional prosthodontic training methods) based on performance skill scores and task completion time, with a significance level set at <.05., Results: The meta-analysis revealed that the utilization of VRIS generally improves students' performance scores (SMD = 1.04; 95% CI, -0.35 to 2.44; I 2  > 50%; p = .13) and reduces task completion time (SMD = -0.03; 95% CI, 1.39-7.72; I 2  > 50%; p = .93). Notably, using VRIS significantly enhanced the performance scores in implant surgery practice (SMD = 0.26; 95% CI, 0.09-0.42; p < .05). Additionally, the VRIS method significantly reduced task completion time in the cavity restorative preparation task (SMD = -1.19; 95% CI, -1.85 to -0.53; p < .05)., Conclusion: Engaging in practice with VRIS has the potential to enhance learning proficiency in prosthodontic education. The advantages associated with VRIS encompass the provision of immediate feedback, decreased task completion time, heightened confidence and motivation, accelerated skill acquisition, improved performance scores, and increased learning engagement., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Decapod Penstylhamaparvovirus 1, commonly known as infectious hypodermal and hematopoietic necrosis virus (IHHNV), remains an economically important viral pathogen for penaeid shrimp aquaculture due to its effects on growth performance. The World Organization for Animal Health (WOAH, Paris, France) recommended methods for the detection of IHHNV include both conventional and real-time PCR. However, published reports and anecdotal evidence suggest the occurrence of non-specific amplifications when testing for IHHNV using the WOAH protocols. Studies were designed to develop a sensitive, robust TaqMan PCR method for detection of IHHNV in the three commercially important penaeid shrimp: Penaeus vannamei, P. monodon and P. stylirostris. We compared the performance of the WOAH-recommended real-time PCR method to several published as well as in-house designed primer/probe sets spanning the entire genome of IHHNV. Our results show that (1) more than one primer/ probe set is needed when testing for the infectious form of IHHNV in all three species of shrimp and (2) primer pairs qIH-Fw/qIH-Rv and 3144F/ 3232R have diagnostic characteristics that would enable IHHNV detection in all three shrimp species. These findings are valuable for a large-scale screening of shrimp using a TaqMan real-time PCR assay., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest with other people or organizations that could inappropriately influence their work., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Microsporidia are emerging intracellular parasites of most known animal phyla in all ecological niches. In shrimp aquaculture, the microsporidium Enterocytozoon hepatopenaei (EHP) is a major cause of concern inflicting tremendous losses to shrimp producers in southeast Asia. During a histopathological examination of Penaeus vannamei samples originating in a country from Latin America presenting slow growth, we observed abnormal nuclei in the epithelial cells of the hepatopancreas. A PCR screening of the samples using DNA isolated from paraffin embedded tissues for the SSU rRNA gene of EHP provided a 149 bp amplicon. In situ hybridization using the SSU rRNA gene probe provided a positive signal in the nuclei instead of the cytoplasm. Sequence analysis of the SSU rRNA gene product revealed a 91.3 %, 89.2 % and 85.4 % sequence identity to Enterocytozoon bieneusi, E. hepatopenaei and Enterospora canceri respectively. Furthermore, phylogenetic analysis revealed the newly discovered microsporidium clustered with E. bieneusi. Considering the intranuclear location of the novel microsporidium and the differences in the sequence of the SSU rRNA, we tentatively consider this parasite a new member of the genus Enterospora sp. The pathogenicity and distribution of the shrimp Enterospora sp. are currently unknown. Our future efforts are focused on the characterization and development of diagnostic tools for this parasite to understand if it acts as an emergent pathogen that might require surveillance to prevent its spread., 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 Elsevier Inc. All rights reserved.)

Viral disease pandemics are a major cause of economic losses in crustacean farming worldwide. While RNA interference (RNAi)-based therapeutics have shown promise at a laboratory scale, without an effective oral delivery platform, RNA-based therapy will not reach its potential against controlling viral diseases in crustaceans. Using a reverse-engineered shrimp RNA virus, Macrobrachium rosenbergii nodavirus (MrNV), we have developed a shrimp viral vector for delivering an engineered RNA cargo. By replacing the RNA-dependent RNA polymerase (RdRp) protein-coding region of MrNV with a cargo RNA encoding green fluorescent protein (GFP) as a proof-of-concept, we generated a replication-incompetent mutant MrNV (ΔRdRp) carrying the GFP RNA cargo resulting in MrNV (ΔRdRp) -GFP. Upon incorporating MrNV (ΔRdRp) -GFP in the diet of the marine Pacific white shrimp ( Penaeus vannamei ), MrNV (ΔRdRp) particles were visualized in hemocytes demonstrating successful vector internalization. Fluorescence imaging of hemocytes showed the expression of GFP protein and the MrNV capsid RNA (RNA2) as well as the incorporated GFP RNA cargo. Detection of cargo RNA in hepatopancreas and pleopods indicated the systemic spread of the viral vector. The quantitative load of both the MrNV RNA2 and GFP RNA progressively diminished within 8 days postadministration of the viral vector, which indicated a lack of MrNV (ΔRdRp) -GFP replication in shrimp. In addition, no pathological hallmarks of the wild-type MrNV infection were detected using histopathology in the target tissue of treated shrimp. The data unequivocally demonstrated the successful engineering of a replication-incompetent viral vector for RNA delivery, paving the way for the oral delivery of antiviral therapeutics in farmed crustaceans., (© The Author(s) 2023. Published by Oxford University Press on behalf of National Academy of Sciences.)

Background: Diabetes mellitus is a chronic progressive metabolic disorder that affects millions of people worldwide. Emerging evidence suggests the important roles of sphingolipid metabolism in diabetes. In particular, sphingosine-1-phosphate (S1P) and S1P receptor 2 (S1PR2) have important metabolic functions and are involved in several metabolic diseases. In diabetes, S1PR2 can effectively preserve β cells and improve glucose/insulin tolerance in high-fat diet induced and streptozotocin (STZ)-induced diabetic mouse models. We previously developed a group of potent and selective S1PR2 ligands and radioligands., Methods: In this study, we continued our efforts and characterized our leading S1PR2 radioligand, [ 11 C]TZ34125, in a STZ-induced diabetic mouse model. [ 11 C]TZ34125 was radiosynthesized in an automated synthesis module and in vitro saturation binding assay was performed using recombinant human S1PR2 membrane. In vitro saturation autoradiography analysis was also performed to determine the binding affinity of [ 11 C]TZ34125 against mouse tissues. Type-1 diabetic mouse model was developed following a single high dose of STZ in C57BL/6 mice. Ex vivo biodistribution was performed to evaluate the distribution and amount of [ 11 C]TZ34125 in tissues. In vitro autoradiography analysis was performed to compare the uptake of [ 11 C]TZ34125 between diabetic and control animals in mouse spleen and pancreas., Results: Our in vitro saturation binding assay using [ 11 C]TZ34125 confirmed [ 11 C]TZ34125 is a potent radioligand to recombinant human S1PR2 membrane with a K d value of 0.9 nM. Saturation autoradiographic analysis showed [ 11 C]TZ34125 has a K d of 67.5, 45.9, and 25.0 nM to mouse kidney, spleen, and liver tissues respectively. Biodistribution study in STZ-induced diabetic mice showed the uptake of [ 11 C]TZ34125 was significantly elevated in the spleen (~2 fold higher) and pancreas (~1.4 fold higher) compared to normal controls. The increased uptake of [ 11 C]TZ34125 was further confirmed using autoradiographic analysis in the spleen and pancreases of STZ-induced diabetic mice, indicating S1PR2 can potentially act as a biomarker of diabetes in pancreases and inflammation in spleen. Future mechanistic analysis and in vivo quantitative assessment using non-invasive PET imaging in large animal model of diabetes is worthwhile., Conclusions: Overall, our data showed an increased uptake of our lead S1PR2-specific radioligand, [ 11 C]TZ34125, in the spleen and pancreases of STZ-induced diabetic mice, and demonstrated [ 11 C]TZ34125 has a great potential for preclinical and clinical usage for assessment of S1PR2 in diabetes and inflammation., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)

The use of three-dimensional (3D) facial scans for facial analysis is increasing in maxillofacial treatment. The aim of this study was to investigate the consistency of two-dimensional (2D) and 3D facial analyses performed by multiple raters. Six men and four women (25-36-year-old) participated in this study. The 2D images of the smiling and resting faces in the frontal and sagittal planes were obtained. The 3D facial and intraoral scans were merged to generate virtual 3D faces. Ten clinicians performed facial analyses by investigating 14 indices of 2D and 3D faces. Intra- and interrater agreements of the results of 2D and 3D facial analyses within and among the participants were evaluated. The intrarater agreement between the 2D and 3D facial analyses varied according to the indices. The highest and lowest agreements were found for the dental crowding index (0.94) and smile line curvature index (0.56) in the frontal plane, and Angle's classification (canine) index (0.98) and occlusal plane angle index (0.55) in the profile plane. In the frontal plane, the interrater agreements were generally higher for the 3D images than for the 2D images, while in the profile plane, the interrater agreements were high in the Angle's classification (canine) index however low in the other indices. Several occlusion-related indices were missing in the 2D images because the posterior teeth were not observed. Esthetic analysis results between 2D and 3D face images can differ according to the evaluation indices. The use of 3D faces is recommended over 2D images to increase the reliability of facial analyses, as it can fully assess both esthetic and occlusion-related indices., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2023 Minsoo Park et al.)

Here we report for the first time a laboratory challenge model for Enterocytozoon hepatopenaei (EHP) to determine the difference of two Specific Pathogen Free (SPF) lines of Penaeus vannamei shrimp. These lines were experimentally challenged using EHP-infected fecal strings as inoculum. Real-time PCR and histopathology assays were performed to confirm EHP infection and evaluate differences in EHP susceptibility in the two genetic lines screened. Although the histopathology of the hepatopancreas tissue showed EHP lesions in both challenged groups, the histological lesions were more pronounced in one of the SPF lines. Quantitative PCR results revealed that animals displaying less hepatocellular damage have lower EHP load compared to animals displaying more pronounced pathological changes. There was no significant difference in final survival at 36 days post-infection in these lines with survival ranging between 80 and 100%. The data showed that mortality as an endpoint metric is not a suitable parameter to determine genetic susceptibility to EHP. Instead, histopathological changes in hepatopancreas, EHP load of the same tissue, and growth retardation would be better metrics to screen EHP susceptibility in P. vannamei. The results show the feasibility of screening genetic lines of P. vannamei for EHP resistance/tolerance using fecal string as an inoculum and, assessing histopathological changes, EHP load, and weight as indicators of resistance., 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 © 2022 Elsevier Inc. All rights reserved.)

Purpose: Accuracy of image matching between resting and smiling facial models is affected by the stability of the reference surfaces. This study aimed to investigate the morphometric variations in subdivided facial units during resting, posed and spontaneous smiling., Materials and Methods: The posed and spontaneous smiling faces of 33 adults were digitized and registered to the resting faces. The morphological changes of subdivided facial units at the forehead (upper and lower central, upper and lower lateral, and temple), nasal (dorsum, tip, lateral wall, and alar lobules), and chin (central and lateral) regions were assessed by measuring the 3D mesh deviations between the smiling and resting facial models. The one-way analysis of variance, Duncan post hoc tests, and Student's t-test were used to determine the differences among the groups (α = .05)., Results: The smallest morphometric changes were observed at the upper and central forehead and nasal dorsum; meanwhile, the largest deviation was found at the nasal alar lobules in both the posed and spontaneous smiles ( P < .001). The spontaneous smile generally resulted in larger facial unit changes than the posed smile, and significant difference was observed at the alar lobules, central chin, and lateral chin units ( P < .001)., Conclusion: The upper and central forehead and nasal dorsum are reliable areas for image matching between resting and smiling 3D facial images. The central chin area can be considered an additional reference area for posed smiles; however, special cautions should be taken when selecting this area as references for spontaneous smiles., (© 2023 The Korean Academy of Prosthodontics.)

Purpose: This study aimed to investigate the accuracy of automatic tooth finish line registration compared to manual registration with regard to various finish line configurations and dental computer-aided design (CAD) software., Methods: Finish line registrations were performed on 15 digital tooth models with different finish line configurations (edge roundness radius = 0 mm, 0.2 mm, and 0.4 mm; edge angle = 30°, 60°, 90°, 120°, and 150°) using automatic and manual methods for designing virtual copings (N = 150). The discrepancies between the registered finish line extracted from the copings and the actual finish line segmented from the digitized tooth model were measured. Three-way analysis of variance (ANOVA) and post-hoc analyses with Bonferroni correction (α = 0.05) were used to analyze the results., Results: The finish line configurations, registration methods, and CAD software interacted with the accuracy of the registered finish line (p = 0.001). The automatic finish line registration method exhibited larger error values than the manual method, especially at high finish line edge roundness and obtuse edge angles for both EXOCAD and R2CAD software (p < 0.001). The difference in dental CAD software affected the registration accuracy in the automatic method (p < 0.001), but not in the manual method (p = 0.676)., Conclusions: Finish line registration errors may occur when the automatic registration method is applied to the indistinct edge of tooth preparation. The accuracy of the automatic finish line registration could differ according to the CAD software program.

Background: The novel concept of immersive 3D augmented reality (AR) surgical navigation has recently been introduced in the medical field. This method allows surgeons to directly focus on the surgical objective without having to look at a separate monitor. In the dental field, the recently developed AR-assisted dental implant navigation system (AR navigation), which uses innovative image technology to directly visualize and track a presurgical plan over an actual surgical site, has attracted great interest., Objective: This study is the first systematic review and meta-analysis study that aimed to assess the accuracy of dental implants placed by AR navigation and compare it with that of the widely used implant placement methods, including the freehand method (FH), template-based static guidance (TG), and conventional navigation (CN)., Methods: Individual search strategies were used in PubMed (MEDLINE), Scopus, ScienceDirect, Cochrane Library, and Google Scholar to search for articles published until March 21, 2022. This study was performed in accordance with the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines and registered in the International Prospective Register of Systematic Reviews (PROSPERO) database. Peer-reviewed journal articles evaluating the positional deviations of dental implants placed using AR-assisted implant navigation systems were included. Cohen d statistical power analysis was used to investigate the effect size estimate and CIs of standardized mean differences (SMDs) between data sets., Results: Among the 425 articles retrieved, 15 articles were considered eligible for narrative review, 8 articles were considered for single-arm meta-analysis, and 4 were included in a 2-arm meta-analysis. The mean lateral, global, depth, and angular deviations of the dental implant placed using AR navigation were 0.90 (95% CI 0.78-1.02) mm, 1.18 (95% CI 0.95-1.41) mm, 0.78 (95% CI 0.48-1.08) mm, and 3.96° (95% CI 3.45°-4.48°), respectively. The accuracy of AR navigation was significantly higher than that of the FH method (SMD=-1.01; 95% CI -1.47 to -0.55; P<.001) and CN method (SMD=-0.46; 95% CI -0.64 to -0.29; P<.001). However, the accuracies of the AR navigation and TG methods were similar (SMD=0.06; 95% CI -0.62 to 0.74; P=.73)., Conclusions: The positional deviations of AR-navigated implant placements were within the safety zone, suggesting clinically acceptable accuracy of the AR navigation method. Moreover, the accuracy of AR implant navigation was comparable with that of the highly recommended dental implant-guided surgery method, TG, and superior to that of the conventional FH and CN methods. This review highlights the possibility of using AR navigation as an effective and accurate immersive surgical guide for dental implant placement., (©Hang-Nga Mai, Van Viet Dam, Du-Hyeong Lee. Originally published in the Journal of Medical Internet Research (https://www.jmir.org), 04.01.2023.)

Statement of Problem: Accurate intraoral preparation of screw channels in cement-retained implant prostheses could be a treatment challenge for tilted or badly positioned implants., Purpose: The purpose of this randomized clinical trial was to evaluate screw channel accuracy for retrieving cement-retained implant prostheses by using a digitally fabricated drilling guide., Material and Methods: Twenty-four participants requiring removal of cement-retained implant prostheses were recruited and consecutively assigned to 1 of 2 groups. In the first group, drilling guides were fabricated by using an intraoral scan, computer-aided design (CAD), and 3D printing to facilitate screw channel preparation (guide group). In the second group, screw channel preparation was based on panoramic radiographs without a drilling guide (freehand group). Screw channel accuracy was evaluated with a 3D analysis and subsequently compared with the ideal shape of the screw channel for linear deviation at coronal and apical levels, angular deviation, surface loss deviation, and volume loss deviation. The ideal shape of the screw-access channel was based on the Ø2.2-mm cylinder image marked along the long axis of the implant. The Mann-Whitney U and Kruskal-Wallis tests were used for statistical analyses (α=.05)., Results: The drilling guide group showed higher dimensional accuracy than the freehand group for angular deviation (9.45 degrees versus 15.69 degrees; P=.014) and surface loss deviation (7.40 mm versus 9.53 mm; P=.001), especially when the implant was tilted more than 30 degrees. The deviation values of the screw channels were not significantly different according to implant tilt in either group (P>.05)., Conclusions: Digitally fabricated drilling guides for intraoral preparation of screw channels in cement-retained implant prostheses can improve accuracy and provide a less-destructive screw channel., (Copyright © 2022 Editorial Council for The Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

The emergence and spread of disease-causing viruses in shrimp aquaculture is not uncommon. Since 2016, unusual mortalities have been affecting the Brazilian shrimp industry and we have associated these unusual mortalities with a novel variant of infectious myonecrosis virus (IMNV). The transcriptome analysis of these diseased shrimp showed an additional divergent viral sequence that we have assigned to the family Solinviviridae . The novel virus has been tentatively termed Penaeus vannamei solinvivirus (PvSV) (GenBank accession: OP265432). The full-length genome of the PvSV is 10.44 kb (excluding the poly A tail) and codes for a polyprotein of 3326 aa. Five conserved domains coding for a helicase, RdRp, calicivirus coat protein, G-patch and tegument protein were identified. The genome organization of the PvSV is similar to other ( Nylan deria fulva virus 1) solinvivirus. A unique feature of this virus that differs from other members of the Solinviviridae is the presence of putative nuclear localization signals. The tissue tropism of this virus is wide, infecting cells of the hepatopancreas, gastrointestinal tract, lymphoid organ and muscle tissue. Another unique feature is that it is the only RNA virus of penaeid shrimp that shows a nuclear localization by in situ hybridization. The PvSV has a wide distribution in Brazil and has been found in the states of Maranhão State (Perizes de Baixo), Piaui State (Mexeriqueira), Ceará State (Camocim, Jaguaruana, Aracati and Alto Santo) and Pará State where it has been detected in coinfections with IMNV. The diagnostic methods developed here (real-time RT-PCR and in situ hybridization) are effective for the detection of the pathogen and should be employed to limit its spread. Furthermore, the identification of the PvSV shows the increasing host range of the relatively new family Solinviviridae .

Objective: This systematic review and meta-analysis aimed to investigate the accuracy of removable partial denture (RPD) frameworks fabricated by computer-aided design/ computer-aided manufacturing (CAD/CAM) systems compared to frameworks produced by conventional casting methods., Methods: A systematic literature search was conducted in electronic databases following the preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines, using formulated Boolean operator for searching articles evaluating the fit accuracy of CAD/CAM-fabricated RPD metal frameworks. The agreement of data collection among the reviewers was confirmed using the Cohen kappa coefficient. The modified methodological index for non-randomized studies (MINORS) scale was used to assess the quality of the included studies. Heterogeneity among studies was evaluated, and meta-analyses with global and subgroup analyses were performed., Results: A total of 208 articles were identified with 18 articles available for the narrative review and 7 studies available for meta-analysis on the 3D-printed frameworks for RPD (the overall Cohen kappa coefficient = 0.944). There was no article on the accuracy of RPD frameworks made by milling method. The meta-analysis revealed that the misfit of frameworks obtained with the CAD/CAM method was significantly higher than that of the conventional lost wax and casting method (SMD = 1.23 μm, 95% CI = 0.3610-2.0981 μm, z = 2.77, P = .0055). Regarding the CAD/CAM techniques used for fabricating RPD metal frameworks, the within subgroup analysis showed that the CAD/CAM-based indirect fabrication method produced frameworks with fit accuracy more similar to the conventional lost-wax and casting method (SMD = 1.15 μm, 95% CI = - 0.06136-2.9192 μm, z = 1.28, P < .01) than the CAD/CAM-based direct fabrication method (SMD = 1.35 μm, 95% CI = -0.2722-2.4381 μm, z = 2.45, P < .01), even though there was no statistical difference between the techniques (P = .8482)., Conclusions: The fit accuracy of RPD metal frameworks fabricated by the CAD/CAM method was lower than that of frameworks fabricated by the conventional casting method, but it was within the clinically acceptable range in dimensional misfit. The indirect metal 3D printing technique based on pattern printing and subsequent casting produced frameworks with fit accuracy more similar to the conventional method than the direct fabrication technique., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Infection with infectious hypodermal and hematopoietic necrosis virus (IHHNV) is a crustacean disease that caused large-scale mortality in Penaeus stylirostris, deformity and growth retardation in Penaeus vannamei and Penaeus monodon. We surveyed the presence of IHHNV in three major shrimp-producing regions in Ecuador, namely Guayas, El Oro, and Esmeralda. The data show that IHHNV is endemic (3.3-100% prevalence) to shrimp farms in these regions. The whole genome sequences of representative circulating IHHNV genotypes in Ecuador and Peru showed that these genotypes formed a separate cluster within the Type II genotypes and were divergent from other geographical isolates of IHHNV originating in Asia, Africa, Australia, and Brazil. In experimental bioassays using specific pathogen-free (SPF) P. vannamei, P. monodon, and P. stylirostris and representative IHHNV isolates from Ecuador and Peru, the virus did not cause any mortality or induce clinical signs in any of the three penaeid species. Although IHHNV-specific Cowdry type A inclusion bodies were histologically detected in experimentally challenged P. vannamei and P. monodon and confirmed by in situ hybridization, no such inclusions were observed in P. stylirostris. Moreover, P. vannamei had the highest viral load, followed by P. monodon and P. stylirostris. Based on IHHNV surveillance data, we conclude that the currently farmed P. vannamei lines in Ecuador are tolerant to circulating IHHNV genotypes. The genome sequence and experimental bioassay data showed that, although the currently circulating genotypes are infectious, they do not induce clinical lesions in the three commercially important penaeid species. These findings suggest a potentially evolving virus-host relationship where circulating genotypes of IHHNV co-exist in equilibrium with P. vannamei raised in Peru and Ecuador., Competing Interests: The authors have declared that no competing interests exist.

A new autonomous water-enabled self-healing coating with antibacterial-agent-releasing capability was developed for the first time by precipitating an aqueous solution of hydrogen-bonded tannic acid (TA) and polyethylene glycol (PEG) (TA: 5 mg/mL; PEG: 5 mg/mL with M W = 100 kDa) to form a smooth, uniform coating layer with an average roughness of 0.688 nm and thickness of 22.3 μm on a polymethyl methacrylate (PMMA) substrate after 10 min of incubation. Our method is cost- and time-efficient, as the hydrophilic coating (water contact angle = 65.1°) forms rapidly, binding strongly to the PMMA substrate (adhesive energy = 83 mJ/m 2 ), without the need for pretreatment or surface modification, and is capable of rapid self-repair (approximately 5 min) through hydrogen bonding in aqueous media. Furthermore, adding 0.5 mg/mL of chlorhexidine acetate (CHX), a commonly used antibacterial agent in dentistry, into the TA-PEG emulsion allowed the release of 2.89 μg/mL of the drug from the coating layer, which is promising for actively inhibiting the vitality and growth of bacteria around PMMA dental restorations. The use of CHX-loaded TA-PEG hydrogen-bonded complexes is highly favorable for the fabrication of an autonomous self-healing biocoating with active antibacterial-agent-releasing capability, which can be applied not only in dentistry but also in other medical fields.

Purpose: This study investigated the accuracy of full-arch intraoral scans obtained by various scan strategies with the segmental scan and merge methods., Materials and Methods: Seventy intraoral scans (seven scans per group) were performed using 10 scan strategies that differed in the segmental scan (1, 2, or 3 segments) and the scanning motion (straight, zigzag, or combined). The three-dimensional (3D) geometric accuracy of scan images was evaluated by comparison with a reference image in an image analysis software program, in terms of the arch shape discrepancies. Measurement parameters were the intermolar distance, interpremolar distance, anteroposterior distance, and global surface deviation. One-way analysis of variance and Tukey honestly significance difference post hoc tests were carried out to compare differences among the scan strategy groups (α = .05)., Results: The linear discrepancy values of intraoral scans were not different among scan strategies performed with the single scan and segmental scan methods. In general, differences in the scan motion did not show different accuracies, except for the intermolar distance measured under the scan conditions of a 3-segmental scan and zigzag motion. The global surface deviations were not different among all scan strategies., Conclusion: The segmental scan and merge methods using two scan parts appear to be reliable as an alternative to the single scan method for full-arch intraoral scans. When three segmental scans are involved, the accuracy of complete arch scan can be negatively affected., (© 2022 The Korean Academy of Prosthodontics.)

Recently, three-dimensional (3D) facial scanning has been gaining popularity in personalized dentistry. Integration of the digital dental model into the 3D facial image allows for a treatment plan to be made in accordance with the patients’ individual needs. The aim of this study was to evaluate the effects of extraoral markers on the accuracy of digital dentofacial integrations. Facial models were generated using smartphone and stereophotogrammetry. Dental models were generated with and without extraoral markers and were registered to the facial models by matching the teeth or markers (n = 10 in each condition; total = 40). Accuracy of the image integration was measured in terms of general 3D position, occlusal plane, and dental midline deviations. The Mann−Whitney U test and two-way analysis of variance were used to compare results among face-scanning systems and matching methods (α = 0.05). As result, the accuracy of dentofacial registration was significantly affected by the use of artificial markers and different face-scanning systems (p < 0.001). The deviations were smallest in stereophotogrammetry with the marker-based matching and highest in smartphone face scans with the tooth-based matching. In comparison between the two face-scanning systems, the stereophotogrammetry generally produced smaller discrepancies than smartphones.

White Feces Syndrome (WFS) is an emergent disease of penaeid shrimp (Penaeus monodon and P. vannamei) that is identified by the presence of floating white fecal strings on pond water in grow-out ponds. Although the clinical manifestations of WFS are well defined, the underling etiology remains obscure. WFS has been associated with several enteric pathogens, including Enterocytozoon hepatopenaei (EHP). The association is based on studies that found areas where WFS has been reported, the prevalence and severity of EHP infection are high. In this study, we describe an experimental reproduction of WFS in P. vannamei pre-infected with EHP and challenged with a unique isolate of Vibrio parahaemolyticus isolated from the gastrointestinal tract of a shrimp displaying WFS. Upon laboratory challenge, shrimp displaying white fecal strings and white discoloration of the gastrointestinal tract were analyzed by histopathology, in-situ hybridization and quantitative PCR. Histological analysis confirmed the lesions of EHP and septic hepatopancreatic necrosis in the hepatopancreas of shrimp exposed to both pathogens. Quantitative PCR showed shrimp infected with both EHP and V. parahaemolyticus had a significantly higher load of EHP compared to shrimp infected with EHP alone. This is the first demonstration of experimental reproduction of WFS under laboratory conditions when animals are infected with EHP and V. parahaemolyticus concurrently. The data revealed a synergistic relation between EHP and V. parahaemolyticus isolate that led to the manifestation of WFS. We propose the gross signs of WFS can be used as an indicator of the presence of EHP infection in association with a particular strain of an enteric Vibrio spp. in countries where EHP is endemic., Competing Interests: The authors have declared that no competing interests exist.

Different stem cell-based strategies, especially induced pluripotent stem cells (iPSCs), have been exploited to regenerate teeth or restore biological and physiological functions after tooth loss. Further research is needed to establish an optimized protocol to effectively differentiate human iPSCs (hiPSCs) into dental epithelial cells (DECs). In this study, various factors were precisely modulated to facilitate differentiation of hiPSCs into DECs, which are essential for the regeneration of functional teeth. Embryoid bodies (EBs) were formed from hiPSCs as embryo-like aggregates, retinoic acid (RA) was used as an early ectodermal inducer, and bone morphogenic protein 4 (BMP4) activity was manipulated. The characteristics of DECs were enhanced and preserved after culture in keratinocyte serum-free medium (K-SFM). The yielded cell population exhibited noticeable DEC characteristics, consistent with the expression of epithelial cell and ameloblast markers. DECs demonstrated odontogenic abilities by exerting an inductive effect on human dental pulp stem cells (hDPSCs) and forming a tooth-like structure with the mouse tooth mesenchyme. Overall, our differentiation protocol provides a practical approach for applying hiPSCs for tooth regeneration., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Induced pluripotent stem cell (iPSC)-based technology provides limitless resources for customized development of organs without any ethical concerns. In theory, iPSCs generated from terminally differentiated cells can be induced to further differentiate into all types of organs that are derived from the embryonic germ layers. Since iPSC reprogramming technology is relatively new, extensive efforts by the researchers have been put together to optimize the protocols to establish in vitro differentiation of human iPSCs (hiPSCs) into various desirable cell types/organs., Highlights: In the present study, we review the potential application of iPSCs as an efficient alternative to primary cells for modulating signal molecules. Furthermore, an efficient culture system that promotes the differentiation of cell lineages and tissue formation has been reviewed. We also summarize the recent studies wherein tissue engineering of the three germ layers has been explored. Particularly, we focus on the current research strategies for iPSC-based tooth regeneration via molecular modulation., Conclusion: In recent decades, robust knowledge regarding the hiPSC-based regenerative therapy has been accumulated, especially focusing on cellular modulation. This review provides the optimization of the procedures designed to regenerate specific organs., Competing Interests: Conflicts of interest The authors declare that there are no conflicts of interest., (Copyright © 2021 Japanese Association for Oral Biology. Published by Elsevier B.V. All rights reserved.)

This study is aimed at assessing the effects of exposure parameters and voxel size for cone-beam computed tomography (CBCT) on the image matching accuracy with an optical dental scan image. CBCT and optical scan images of a dry human mandible were obtained. Different CBCT settings were used: tube voltage, 60, 80, and 100 kVp; tube current, 6 and 8 mA; and voxel size, 100, 200, and 300  μ m. Image matching between the CBCT and optical scan images was performed using implant planning software by dental professionals ( n = 18). The image matching accuracy in each combination of CBCT settings was evaluated by assessing the linear discrepancy between the three-dimensionally reconstructed radiological image and the registered optical scan image using an image analysis software program. The Kruskal-Wallis test and a post hoc Mann-Whitney U test with Bonferroni correction were used to compare the accuracy of image registration between the groups ( α = 0.05). Overall, the image matching accuracy was not significantly different between tube voltage and current settings; however, significantly higher image registration errors were found at the combination of 100 kVp tube voltage/8 mA tube current ( F = 8.44, P < 0.001). Changes in voxel sizes did not significantly interfere with the image registration results. No interaction was found among voltage, current, and voxel size in terms of image registration accuracy ( F = 2.022, P = 0.091). Different exposure parameter settings in tube voltage and tube current did not significantly influence the image matching accuracy between CBCT and optical dental scan images; however, a high radiation dose could be inappropriate. The image matching accuracy was not significantly affected by changing the voxel sizes of CBCT., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2021 Hang-Nga Mai and Du-Hyeong Lee.)

Acute hepatopancreatic necrosis disease (AHPND) is currently the most important bacterial disease of shrimp that has caused enormous losses to the shrimp industry worldwide. The causative agent of AHPND are Vibrio spp. Carrying plasmids containing the pirA and pirB genes which encode binary toxins, PirAB. Currently, AHPND is mostly diagnosed by PCR-based platforms which require the use of sophisticated laboratory instrumentation and are not suitable for a point-of-care diagnostics. Therefore, the availability of an alternative method based on isothermal amplification would be suitable for AHPND detection outside a laboratory setting and extremely useful at a pond side location. Isothermal amplification is based on the nucleic acid amplification at a single temperature and does not require the use of a thermal cycler. In this study, we developed an isothermal Recombinase Polymerase Amplification (RPA) assay for AHPND detection targeting both pirA and pirB genes, simultaneously and evaluated the specificity and sensitivity of the assay. The assay could detect AHPND without any cross-reaction with other microbial pathogens and Specific Pathogen Free (SPF) shrimp. The limit of detection of the assay was 5 copies of pirAB genes. To evaluate the reliability of the assay in detecting AHPND, DNA from Penaeus vannamei shrimp displaying acute and chronic infection were analyzed by the RPA assay and the results were compared with SYBR Green real-time PCR assay. While there was a 100% conformity between the two assay while detecting acute phase infection, RPA appeared to be more sensitive in detecting chronic phase infection. The data suggest that RPA assay described here would be a reliable method in detecting AHPND outside a standard laboratory setting., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Acute hepatopancreatic necrosis disease (AHPND) is a lethal disease in marine shrimp that has caused large-scale mortalities in shrimp aquaculture in Asia and the Americas. The etiologic agent is a pathogenic Vibrio sp. carrying binary toxin genes, pir A and pir B in plasmid DNA. Developing AHPND tolerant shrimp lines is one of the prophylactic approaches to combat this disease. A selected genetic line of Penaeus vannamei was found to be tolerant to AHPND during screening for disease resistance. The mRNA expression of twelve immune and metabolic genes known to be involved in bacterial pathogenesis were measured by quantitative RT-PCR in two populations of shrimp, namely P1 that showed susceptibility to AHPND, and P2 that showed tolerance to AHPND. Among these genes, the mRNA expression of chymotrypsin A (ChyA) and serine protease (SP), genes that are involved in metabolism, and crustin-P (CRSTP) and prophenol oxidase activation system 2 (PPAE2), genes involved in bacterial pathogenesis in shrimp, showed differential expression between the two populations. The differential expression of these genes shed light on the mechanism of tolerance against AHPND and these genes can potentially serve as candidate markers for tolerance/susceptibility to AHPND in P. vannamei . This is the first report of a comparison of the mRNA expression profiles of AHPND tolerant and susceptible lines of P. vannamei ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Mai, Caro, Cruz-Flores, White and Dhar.)

Hypoxia is a state of inadequate supply of oxygen. Increasing evidence indicates that a hypoxic environment is strongly associated with abnormal organ development. Oxygen nanobubbles (ONBs) are newly developed nanomaterials that can deliver oxygen to developing tissues, including hypoxic cells. However, the mechanisms through which nanobubbles recover hypoxic tissues, such as developing tooth germs remain to be identified. In this study, tooth germs were cultured in various conditions: CO 2 chamber, hypoxic chamber, and with 20% ONBs for 3 h. The target stages were at the cap stage (all soft tissue) and bell stage (hard tissue starts to form). Hypoxic tooth germs were recovered with 20% ONBs in the media, similar to the tooth germs incubated in a CO 2 chamber (normoxic condition). The tooth germs under hypoxic conditions underwent apoptosis both at the cap and bell stages, and ONBs rescued the damaged tooth germs in both the cap and bell stages. Using kidney transplantation for hard tissue formation in vivo , amelogenesis and dentinogenesis imperfecta in hypoxic conditions at the bell stage were rescued with ONBs. Furthermore, glucose uptake by tooth germs was highly upregulated under hypoxic conditions, and was restored with ONBs to normoxia levels. Our findings indicate that the strategies to make use of ONBs for efficient oxygen targeted delivery can restore cellular processes, such as cell proliferation and apoptosis, glucose uptake, and hypomineralization in hypoxic environments., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kim, Lee, Yoon, Lee, Mai, Ida-Yonemochi, Choi and Jung.)

Background: Enterocytozoon hepatopenaei (EHP) is an enteric pathogen that affects Penaeus vannamei and Penaeus monodon shrimp in many SE Asian countries. In the western hemisphere, EHP was reported for the first time in 2016 in farmed P. vannamei in Venezuela. Anecdotal evidence suggests that EHP is more prevalent in grow-out ponds where the salinity is high (> 15 parts per thousand (ppt)) compared to grow-out ponds with low salinities (< 5 ppt). Considering that P. vannamei is an euryhaline species, we were interested in knowing if EHP can propagate in P. vannamei in low salinities., Results: In this study, we described an experimental infection using fecal strings as a source inoculum. Specific Pathogen Free (SPF) P. vannamei were maintained at three different salinities (2 ppt, 15 ppt, and 30 ppt) while continuously challenged using feces from known EHP-infected P. vannamei over a period of 3 weeks. The fecal strings, used as a source of EHP inocula in the challenges, was sufficient to elicit an infection in shrimp maintained at the three salinities. The infectivity of EHP in shrimp reared at 2 ppt, 15 ppt, and 30 ppt salinities was confirmed by PCR and histopathology. The prevalence and the severity of the EHP infection was higher at 30 ppt than at 2 ppt and 15 ppt., Conclusion: The data suggests that fecal strings are a reliable source of EHP inoculum to conduct experimental challenges via the fecal-oral route. An EHP infection can occur at a salinity as low as 2 ppt, however, the prevalence and the severity of the EHP infection is higher at a salinity of 30 ppt.

Glutathione peroxidase (GPx) acts in co-ordination with other signaling molecules to exert its own antioxidant role. We have demonstrated the protective effects of GPx,/GPx-1, a selenium-dependent enzyme, on various neurodegenerative disorders (i.e., Parkinson's disease, Alzheimer's disease, cerebral ischemia, and convulsive disorders). In addition, we summarized the recent findings indicating that GPx-1 might play a role as a neuromodulator in neuropsychiatric conditions, such as, stress, bipolar disorder, schizophrenia, and drug intoxication. In this review, we attempted to highlight the mechanistic scenarios mediated by the GPx/GPx-1 gene in impacting these neurodegenerative and neuropsychiatric disorders, and hope to provide new insights on the therapeutic interventions against these disorders., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Davidson's-fixed paraffin-embedded (DFPE) shrimp tissue are a priceless biological resource for pathogen discovery and evolutionary studies for aquaculture disease diagnostic laboratories worldwide. Nucleic acids extracted from DFPE tissues are often not adequate for most downstream molecular analysis due to fragmentation and chemical modifications. In this study, next generation sequencing (NGS) was used to reconstruct the complete genome of three geographical isolates (Belize, Venezuela and Hawaii) of a ~10 kb length RNA virus of shrimp, Taura syndrome virus (TSV), from DFPE tissues that have been archived for 15 years. Phylogenetic analyses showed that TSV isolates from Belize, Venezuela and Hawaii formed well supported clusters with homologous isolates from the corresponding regions submitted in the GenBank database. This is the first study to demonstrate the utility of archived tissue samples for identification of RNA viruses and evolutionary studies involving a viral disease in crustaceans and opens an avenue for expediting pathogen discovery., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Hepatitis C virus (HCV) life cycle is highly dependent on cellular proteins for viral propagation. In order to identify the cellular factors involved in HCV propagation, we previously performed a protein microarray assay using the HCV nonstructural 5A (NS5A) protein as a probe. Of ∼9,000 human cellular proteins immobilized in a microarray, adenosylhomocysteinase like 1 (AHCYL1) was among 90 proteins identified as NS5A interactors. Of these candidates, AHCYL1 was selected for further study. In the present study, we verified the physical interaction between NS5A and AHCYL1 by both in vitro pulldown and coimmunoprecipitation assays. Furthermore, HCV NS5A interacted with endogenous AHCYL1 in Jc1-infected cells. Both NS5A and AHCYL1 were colocalized in the cytoplasmic region in HCV-replicating cells. siRNAmediated knockdown of AHCYL1 abrogated HCV propagation. Exogenous expression of the siRNA-resistant AHCYL1 mutant, but not of the wild-type AHCYL1, restored HCV protein expression levels, indicating that AHCYL1 was required specifically for HCV propagation. Importantly, AHCYL1 was involved in the HCV internal ribosome entry site-mediated translation step of the HCV life cycle. Finally, we demonstrated that the proteasomal degradation pathway of AHCYL1 was modulated by persistent HCV infection. Collectively, these data suggest that HCV may modulate the AHCYL1 protein to promote viral propagation.

This study evaluated the effects of different matching methods on the accuracy of dentofacial integration in stereophotogrammetry and smartphone face-scanning systems. The integration was done (N = 30) with different matching areas ( n = 10), including teeth image only (TO), perioral area without markers (PN) and with markers (PM). The positional accuracy of the integrated models was assessed by measuring the midline linear deviations and incisal line canting between the experimental groups and laser scanner-based reference standards. Kruskal-Wallis and Mann-Whitney U tests were used for statistical analyses (α = 0.05). The PM method exhibited the smallest linear deviations in both systems; while the highest deviations were found in the TO in stereophotogrammetry; and in PN in smartphone. For the incisal line canting; the canting degree was the lowest in the PM method; followed by that in the TO and the PN in both systems. Although stereophotogrammetry generally exhibited higher accuracy than the smartphone; the two systems demonstrated no significant difference when the perioral areas were used for matching. The use of perioral scans with markers enables accurate dentofacial image integration; however; cautions should be given on the accuracy of the perioral image obtained without the use of markers.

The use of three-dimensional face-scanning systems to obtain facial models is of increasing interest, however, systematic assessments of the reliability of portable face-scan devices have not been widely conducted. Therefore, a systematic review and meta-analysis were performed considering the accuracy of facial models obtained by portable face-scanners in comparison with that of those obtained by stationary face-scanning systems. A systematic literature search was conducted in electronic databases following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines for articles published from 1 January 2009 to 18 March 2020. A total of 2806 articles were identified, with 21 articles available for the narrative review and nine studies available for meta-analysis. The meta-analysis revealed that the accuracy of the digital face models generated by the portable scanners was not significantly different from that of the stationary face-scanning systems (standard mean difference (95% confidence interval) = -0.325 mm (-1.186 to 0.536); z = -0.74; p = 0.459). Within the comparison of the portable systems, no statistically significant difference was found concerning the accuracy of the facial models among scanning methods ( p = 0.063). Overall, portable face-scan devices can be considered reliable for obtaining facial models. However, caution is needed when applying face-scanners with respect to scanning device settings, control of involuntary facial movements, landmark and facial region identifications, and scanning protocols.

Statement of Problem: Implant placement errors have been reported in guided surgeries because of movement of the guide template during implant placement. With a completely limiting guide design with high restrictions, guide template stabilization is essential to minimize mobility during the drilling process., Purpose: The purpose of this in vitro study was to evaluate the effects of supporting conditions and the use of an anchor microscrew on the seating and functional stabilization of a computer-aided design and computer-aided manufacturing (CAD-CAM) implant surgical guide with a completely limiting design., Material and Methods: Twelve implant surgical guide templates were fabricated under different supporting conditions: tooth-mucosa, tooth-tooth, tooth-screw, and screw-screw (n=3 per group). In the tooth-screw and screw-screw groups, anchor microscrews and anchor caps were used to fix the guide templates. The seating stability of the templates was evaluated by using a micromovement assessment. The functional stability of the templates was analyzed during the drilling process by using a motion tracking method. One-way analysis of variance and the Tukey HSD post hoc test were conducted to compare the differences among the groups (α=.05)., Results: The tooth-tooth, tooth-screw, and screw-screw groups exhibited similar seating stability, which was significantly more stable than that of the tooth-mucosa group (P<.001). In terms of functional stability, the tooth-mucosa group reported the highest positional deviation and motion magnitude, whereas low values were found in the tooth-screw and screw-screw groups (P<.001)., Conclusions: The use of anchor microscrews can increase the seating and functional stability of CAD-CAM guide templates during the drilling process for implant placement., (Copyright © 2020 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

We demonstrated that the gene of glutathione peroxidase-1 (GPx-1), a major antioxidant enzyme, is a potential protectant against the neurotoxicity and conditioned place preference induced by cocaine. Because the sigma (σ)-1 receptor is implicated in cocaine-induced drug dependence, we investigated whether the GPx-1 gene modulates the σ-1 receptor in the behavioral sensitization induced by cocaine. Cocaine-induced behavioral sensitization was more pronounced in GPx-1 knockout (KO) than wild-type (WT) mice and was less pronounced in GPx-1 overexpressing transgenic (GPx-1 TG) than non-TG mice. Cocaine treatment significantly enhanced the oxidative burden and reduced the GSH levels in the striatum of WT, GPx-1 KO, and non-TG mice but not in that of GPx-1 TG mice. In addition, cocaine significantly increased the nuclear translocation, its DNA binding activity of nuclear factor erythroid-2-related factor 2 (Nrf2) as well as the mRNA expression of γ-glutamylcysteine (GCL). The genetic depletion of GPx-1 inhibited the Nrf2-related glutathione system, whereas the genetic overexpression of GPx-1 activated this system against behavioral sensitization. BD1047, a σ-1 receptor antagonist, and U0126, an ERK inhibitor significantly induced the Nrf2-related antioxidant potential against behavioral sensitization. Unlike BD1047, U0126 did not affect the cocaine-induced σ-1 receptor immunoreactivity, suggesting that the σ-1 receptor is an upstream molecule for ERK signaling. Importantly, BD1047 and U0126 failed to affect the σ-1 receptor immunoreactivity and ERK phosphorylation induced by cocaine in GPx-1 TG mice. Our results suggest that GPx-1 is a critical mediator for the attenuation of cocaine-induced behavioral sensitization via modulating σ-1 receptor-mediated ERK activation by the induction of the Nrf2-related system., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: The accurate assessment and acquisition of facial anatomical information significantly contributes to enhancing the reliability of treatments in dental and medical fields, and has applications in fields such as craniomaxillofacial surgery, orthodontics, prosthodontics, orthopedics, and forensic medicine. Mobile device-compatible 3D facial scanners have been reported to be an effective tool for clinical use, but the accuracy of digital facial impressions obtained with the scanners has not been explored., Objective: We aimed to review comparisons of the accuracy of mobile device-compatible face scanners for facial digitization with that of systems for professional 3D facial scanning., Methods: Individual search strategies were employed in PubMed (MEDLINE), Scopus, Science Direct, and Cochrane Library databases to search for articles published up to May 27, 2020. Peer-reviewed journal articles evaluating the accuracy of 3D facial models generated by mobile device-compatible face scanners were included. Cohen d effect size estimates and confidence intervals of standardized mean difference (SMD) data sets were used for meta-analysis., Results: By automatic database searching, 3942 articles were identified, of which 11 articles were considered eligible for narrative review, with 6 studies included in the meta-analysis. Overall, the accuracy of face models obtained using mobile device-compatible face scanners was significantly lower than that of face models obtained using professional 3D facial scanners (SMD 3.96 mm, 95% CI 2.81-5.10 mm; z=6.78; P<.001). The difference between face scanning when performed on inanimate facial models was significantly higher (SMD 10.53 mm, 95% CI 6.29-14.77 mm) than that when performed on living participants (SMD 2.58 mm, 95% CI 1.70-3.47 mm, P<.001, df=12.94)., Conclusions: Overall, mobile device-compatible face scanners did not perform as well as professional scanning systems in 3D facial acquisition, but the deviations were within the clinically acceptable range of <1.5 mm. Significant differences between results when 3D facial scans were performed on inanimate facial objects and when performed on the faces of living participants were found; thus, caution should be exercised when interpreting results from studies conducted on inanimate objects., (©Hang-Nga Mai, Du-Hyeong Lee. Originally published in the Journal of Medical Internet Research (http://www.jmir.org), 23.10.2020.)

Polymers are the most commonly used material for three-dimensional (3D) printing in dentistry; however, the high porosity and water absorptiveness of the material adversely influence biofilm formation on the surface of the 3D-printed dental prostheses. This study evaluated the effects of a newly developed chlorhexidine (CHX)-loaded polydimethylsiloxane (PDMS)-based coating material on the surface microstructure, surface wettability and antibacterial activity of 3D-printing dental polymer. First, mesoporous silica nanoparticles (MSN) were used to encapsulate CHX, and the combination was added to PDMS to synthesize the antibacterial agent-releasing coating substance. Then, a thin coating film was formed on the 3D-printing polymer specimens using oxygen plasma and thermal treatment. The results show that using the coating substance significantly reduced the surface irregularity and increased the hydrophobicity of the specimens. Remarkably, the culture media containing coated specimens had a significantly lower number of bacterial colony formation units than the noncoated specimens, thereby indicating the effective antibacterial activity of the coating.

Vibrio parahaemolyticus carrying binary toxin genes, pir AB, is one of the etiological agents causing acute hepatopancreatic necrosis disease (AHPND) in shrimp. This disease has emerged recently as a major threat to shrimp aquaculture worldwide. During a routine PCR screening of AHPND-causing V. parahaemolyticus strains, an isolate tested PCR positive for pirB (R13) and another isolate tested positive for both the pir A and pir B (R14) genes. To evaluate the pathogenicity of these isolates, specific pathogen-free (SPF) Penaeus vannamei were experimentally challenged. For both R13 and R14 isolates, the final survival rate was 100% at termination of the challenge, whereas the final survival with the AHPND-causing V. parahaemolyticus was 0%. The nucleotide sequence of the plasmid DNA carrying the binary toxin genes revealed that R13 contains a deletion of the entire pirA gene whereas R14 contains the entire coding regions of both pir A and pir B genes. However, R14 possesses an insertion upstream of the pir A gene. In R14, mRNA for both pir A and pir B genes could be detected but no cognate proteins. This shows that the genome of AHPND-causing V. parahaemolyticus is highly plastic and, therefore, detection of the pir A and pir B genes alone by DNA-PCR is insufficient as a diagnostic test for AHPND.

Purpose: The image registration of optical scans to radiographic images is essential for performing computer-guided implant surgery. This study aimed to evaluate the effect of different image matching conditions on the accuracy of image registration for computer-guided implant surgery in completely edentulous jaws., Materials and Methods: The optical scan image of a completely edentulous study model was registered to the respective cone-beam computed tomography data using three different image matching conditions: small point (SP), large point (LP), and entire surface (ES). For the SP and LP groups, gutta-percha markers (1.0 and 3.0 mm in diameter) were attached to a base template, and a radiopaque impression material was relined on the intaglio surface of template in the ES group. Image registration was performed by 20 operators in the images obtained from each group at an interval of 2 weeks (n = 20 in each group), and the registration accuracy was assessed by calculating the aligned position of the edentulous arch image. One-way analysis of variance with Tukey post hoc tests was used to compare the results among the groups (α = 0.05)., Results: The mean registration error was significantly larger in the SP group (0.52 ± 0.19 mm) than in the LP group (0.29 ± 0.08 mm) and ES group (0.27 ±0.06 mm) (F = 24.689, p < 0.001). No difference was found between the LP and ES groups. The image matching discrepancy was more homogeneously distributed on the arch in the ES group than in the other groups., Conclusion: The accuracy of image registration is affected by the size of the congruent area shown in the optical scan and radiographic images. The entire surface-based matching method is more accurate as compared to the small point-based matching method in the image registration for implant planning in full edentulous jaws., (© 2020 by the American College of Prosthodontists.)

An acute hepatopancreatic necrosis disease (AHPND) causes serious losses to the global shrimp industry. The etiologic agent of AHPND is Vibrio spp. carrying a large plasmid which encodes a binary toxin, PirAB. Currently, AHPND is diagnosed by PCR based methods that detect the presences of both pirA and pirB genes. However, the bacterial strains containing the pirA and pirB genes do not always express the binary toxin, resulting in mis-estimation of the virulence of bacterial strains containing pirA and pirB genes. Thus, the immuno based assay (i.e. ELISA) is a promising approach to detect PirA Vp and PirB Vp . In the present study, a total of forty monoclonal antibodies clones (mAb) against PirA Vp (20 mAbs) and PirB Vp (20 mAbs) were screened by western blot analysis to select four mAb clones that show the strongest immunoreactivity in indirect ELISA (iELISA). The four selected mAbs (i.e. 1B9 and 5E9 against PirA Vp ; 7B7 and 7B9 against PirB Vp ) detected specifically Vibrio spp. causing AHPND. In addition, four selected mAbs were able to detect either PirA Vp or PirB Vp down to 0.008 ng/μl. A double blind assay using thirty AHPND-infected and six SPF shrimp Penaeus vannamei were analyzed by iELISA to determine the detection sensitivity of the assay. The results showed that iELISA was able to accurately detect 29 out of 30 AHPND infected shrimp. These finding indicated that iELISA is a reliable method to detect PirA Vp and PirB Vp toxins in infected shrimp and will be a useful tool in AHPND diagnosis and in studying the role of binary toxins in AHPND pathogenesis., 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 © 2020 Elsevier B.V. All rights reserved.)

Statement of Problem: Accurate image matching of the scan with the design of the scan body is essential to replicate the actual implant position in the software program. In situations where the scan part of the scan body is partially embedded in the gingival tissue, the influence of the scan image deficiency on the accuracy of image matching has not been clarified., Purpose: The purpose of this in vitro study was to examine the effects of scan body exposure and different operators on the accuracy of image matching of the scan body., Material and Methods: Six groups with different scan body exposures (fully exposed, 0.5 mm, 1.0 mm, 1.5 mm, 2.0 mm, and 2.5 mm less exposed) were digitized, and the scan images were transferred to a CAD software program, where the design of the scan body was matched to the scan image of each group. Based on the position of the scan body design, a virtual implant was created. The image matching process was performed 7 times for each group by 2 operators (N=84). The linear and angular deviations of the virtual implants were analyzed 3-dimensionally. Two-way ANOVA, equivalence, and concordance correlation coefficient statistics were used to verify the effects of scan body exposure and operator on the image matching., Results: As the exposure of the scan body was reduced, the deviations in implant positioning were significantly increased (P<.001). The concordance correlation coefficient indicated strong agreement between the 2 operators., Conclusions: Reduced exposure of the scan body significantly influenced the accuracy of implant positioning in the software program. Operator differences may not affect the accuracy of scan body image matching., (Copyright © 2020 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Formalin-fixed paraffin-embedded (FFPE) tissues are a priceless resource for diagnostic laboratories worldwide. However, DNA extracted from these tissues is often not optimal for most downstream molecular analysis due to fragmentation and chemical modification. In this study, the complete genome of white spot syndrome virus (WSSV) was reconstructed from ~ 2-year-old archived Davidson's-fixed paraffin-embedded (DFPE) shrimp tissue using Next Generation Sequencing (NGS). A histological analysis was performed on archived DFPE shrimp tissue and a sample showing a high level of WSSV infection was selected for molecular analysis. The viral infection was further confirmed by molecular methods. DNA isolated from DFPE and fresh frozen (FF) tissues were sequenced by NGS. The complete genome reconstruction of WSSV (~ 305 kbp) was achieved from both DFPE and FF tissue. Single nucleotide polymorphisms, insertion and deletions were compared between the genomes. Thirty-eight mutations were identified in the WSSV genomes from the DFPE and FF that differed from the reference genome. This is the first study that has successfully sequenced the complete genome of a virus of over 300 kbp from archival DFPE tissue. These findings demonstrate that DFPE shrimp tissue represents an invaluable resource for prospective and retrospective studies, evolutionary studies and opens avenues for pathogen discovery.