Active N6-Methyladenine Demethylation by DMAD Regulates Gene Expression by Coordinating with Polycomb Protein in Neurons.

A ten-eleven translocation (TET) ortholog exists as a DNA N6-methyladenine (6mA) demethylase (DMAD) in Drosophila. However, the molecular roles of 6mA and DMAD remain unexplored. Through genome-wide 6mA and transcriptome profiling in Drosophila brains and neuronal cells, we found that 6mA may epigenetically regulate a group of genes involved in neurodevelopment and neuronal functions. Mechanistically, DMAD interacts with the Trithorax-related complex protein Wds to maintain active transcription by dynamically demethylating intragenic 6mA. Accumulation of 6mA by depleting DMAD coordinates with Polycomb proteins and contributes to transcriptional repression of these genes. Our findings suggest that active 6mA demethylation by DMAD plays essential roles in fly CNS by orchestrating through added epigenetic mechanisms.

Identification of PSMB5 as a genetic modifier of fragile X-associated tremor/ataxia syndrome.

Fragile X­associated tremor/ataxia syndrome (FXTAS) is a debilitating late-onset neurodegenerative disease in premutation carriers of the expanded CGG repeat in FMR1 that presents with a spectrum of neurological manifestations, such as gait ataxia, intention tremor, and parkinsonism [P. J. Hagerman, R. J. Hagerman, Ann. N. Y. Acad. Sci. 1338, 58­70 (2015); S. Jacquemont et al., JAMA 291, 460­469 (2004)]. Here, we performed whole-genome sequencing (WGS) on male premutation carriers (CGG55­200) and prioritized candidate variants to screen for candidate genetic modifiers using a Drosophila model of FXTAS. We found 18 genes that genetically modulate CGG-associated neurotoxicity in Drosophila, such as Prosbeta5 (PSMB5), pAbp (PABPC1L), e(y)1 (TAF9), and CG14231 (OSGEPL1). Among them, knockdown of Prosbeta5 (PSMB5) suppressed CGG-associated neurodegeneration in the fly as well as in N2A cells. Interestingly, an expression quantitative trait locus variant in PSMB5, PSMB5rs11543947-A, was found to be associated with decreased expression of PSMB5 and delayed onset of FXTAS in human FMR1 premutation carriers. Finally, we demonstrate evidence that PSMB5 knockdown results in suppression of CGG neurotoxicity via both the RAN translation and RNA-mediated toxicity mechanisms, thereby presenting a therapeutic strategy for FXTAS.

Dysregulated mitochondrial and cytosolic tRNA m1A methylation in Alzheimer's disease.

RNA modifications affect many aspects of RNA metabolism and are involved in the regulation of many different biological processes. Mono-methylation of adenosine in the N1 position, N1-methyladensoine (m1A), is a reversible modification that is known to target rRNAs and tRNAs. m1A has been shown to increase tRNA structural stability and induce correct tRNA folding. Recent studies have begun to associate the dysregulation of epitranscriptomic control with age-related disorders such as Alzheimer's disease. Here, we applied the newly developed m1A-quant-seq approach to map the brain abundant m1A RNA modification in the cortex of an Alzheimer's disease mouse model, 5XFAD. We observed hypomethylation in both mitochondrial and cytosolic tRNAs in 5XFAD mice compared with wild type. Furthermore, the main enzymes responsible for the addition of m1A in mitochondrial (TRMT10C, HSD17B10) and cytosolic tRNAs (TRMT61A) displayed decreased expression in 5XFAD compared with wild-type mice. Knockdown of these enzymes results in a more severe phenotype in a Drosophila tau model, and differential m1A methylation is correlated with differences in mature mitochondrial tRNA expression. Collectively, this work suggests that hypo m1A modification in tRNAs may play a role in Alzheimer's disease pathogenesis.

Effects of Banhabaekchulcheonma-Tang on Brain Injury and Cognitive Function Impairment Caused by Bilateral Common Carotid Artery Stenosis in a Mouse Model.

Vascular dementia (VD) is the second most prevalent dementia type, with no drugs approved for its treatment. Here, the effects of Banhabaekchulcheonma-Tang (BBCT) on ischemic brain injury and cognitive function impairment were investigated in a bilateral carotid artery stenosis (BCAS) mouse model. Mice were divided into sham-operated, BCAS control, L-BBCT (40 ml/kg), and H-BBCT (80 ml/kg) groups. BBCT's effects were characterized using the Y-maze test, novel object recognition test (NORT), immunofluorescence staining, RNA sequencing, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) analyses. The NORT revealed cognitive function improvement in the H-BBCT group, while the Y-maze test revealed no significant difference among the four groups. The CD68+ microglia and GFAP+ astrocyte numbers were reduced in the H-BBCT group. Furthermore, H-BBCT treatment restored the dysregulation of gene expression caused by BCAS. The major BBCT targets were predicted to be cell division cycle protein 20 (CDC20), Epidermal growth factor (EGF), and tumor necrosis factor receptor-associated factor 1 (TRAF1). BBCT regulates the neuroactive ligand-receptor interaction and neuropeptide signaling pathways, as predicted by KEGG and GO analyses, respectively. BBCT significantly improved cognitive impairment in a BCAS mouse model by inhibiting microglial and astrocyte activation and regulating the expression of CDC20, EGF, TRAF1, and key proteins in the neuroactive ligand-receptor interaction and neuropeptide signaling pathways.

Transition of allele-specific DNA hydroxymethylation at regulatory loci is associated with phenotypic variation in monozygotic twins discordant for psychiatric disorders.

BACKGROUND: Major psychiatric disorders such as schizophrenia (SCZ) and bipolar disorder (BPD) are complex genetic mental illnesses. Their non-Mendelian features, such as those observed in monozygotic twins discordant for SCZ or BPD, are likely complicated by environmental modifiers of genetic effects. 5-Hydroxymethylcytosine (5hmC) is an important epigenetic mark in gene regulation, and whether it is linked to genetic variants that contribute to non-Mendelian features remains largely unexplored. METHODS: We combined the 5hmC-selective chemical labeling method (5hmC-seq) and whole-genome sequencing (WGS) analysis of peripheral blood DNA obtained from monozygotic (MZ) twins discordant for SCZ or BPD to identify allelic imbalances in hydroxymethylome maps, and examined association of allele-specific hydroxymethylation (AShM) transition with disease susceptibility based on Bayes factors (BF) derived from the Bayesian generalized additive linear mixed model. We then performed multi-omics integrative analysis to determine the molecular pathogenic basis of those AShM sites. We finally employed luciferase reporter, CRISPR/Cas9 technology, electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP), PCR, FM4-64 imaging analysis, and RNA sequencing to validate the function of interested AShM sites in the human neuroblastoma SK-N-SH cells and human embryonic kidney 293T (HEK293T) cells. RESULTS: We identified thousands of genetic variants associated with AShM imbalances that exhibited phenotypic variation-associated AShM changes at regulatory loci. These AShM marks showed plausible associations with SCZ or BPD based on their effects on interactions among transcription factors (TFs), DNA methylation levels, or other epigenomic marks and thus contributed to dysregulated gene expression, which ultimately increased disease susceptibility. We then validated that competitive binding of POU3F2 on the alternative allele at the AShM site rs4558409 (G/T) in PLLP-enhanced PLLP expression, while the hydroxymethylated alternative allele, which alleviated the POU3F2 binding activity at the rs4558409 site, might be associated with the downregulated PLLP expression observed in BPD or SCZ. Moreover, disruption of rs4558409 promoted neural development and vesicle trafficking. CONCLUSION: Our study provides a powerful strategy for prioritizing regulatory risk variants and contributes to our understanding of the interplay between genetic and epigenetic factors in mediating SCZ or BPD susceptibility.

Comparative assessment of marginal and internal gaps of cast-free monolithic zirconia crowns fabricated from 2 intraoral scanners: A prospective, double-blind, randomized clinical trial.

STATEMENT OF PROBLEM: Despite the introduction of intraoral scanners (IOSs) with dual camera triangulation, only a few comparative clinical studies have evaluated their clinical performances in the digital workflow for cast-free restorations. PURPOSE: The purpose of this clinical trial was to assess the clinical efficacy of 2 different technology-based IOSs by evaluating the marginal and internal gaps of cast-free monolithic zirconia crowns fabricated by using a fully digital workflow. MATERIAL AND METHODS: A prospective randomized clinical trial was conducted in 35 participants requiring a single-unit restoration. One crown was fabricated from the scan data obtained with a confocal microscopy-based IOS (Group T), while the other was made with the scan data obtained from an IOS using dual camera triangulation (Group I). A replica technique was used to assess the marginal and internal gaps. The buccolingual and mesiodistal cross-sections were measured, and noninferiority trials were performed. RESULTS: A total of 39 teeth from 35 participants were restored with a single-unit crown. The marginal and axial wall gaps of the crowns in Group I was not inferior to that of the crowns in Group T (upper limit confidence interval [CI] <30). In contrast, the gap of the crowns at the line angle in Group T was inferior to that of the crowns in Group I (lower limit CI <-30). From an occlusal space perspective, the gap of the crowns in Group I was inferior to that of the crowns in Group T (upper limit CI >30). Twenty-five crowns were selected from Group I, and 14 crowns were selected from Group T for definitive placement. CONCLUSIONS: The marginal gap of the crown fabricated by using the scan data obtained from the dual camera triangulation-based IOS was noninferior to that obtained from the confocal microscopy-based IOS and was within the clinically applicable limit.

Sleeve insert scan body to predict implant placement position by using implant surgical guides: A dental technique.

In studies that assessed the accuracy of implant surgical guides, evaluations were based on the placement position of the implant by using a manufactured surgical guide. However, such assessments could involve errors that may occur during implant placement. Therefore, evaluating the 3-dimensional accuracy of the fabrication of the implant surgical guide itself is not enough. In the evaluation method described in this article, location-related information is obtained by connecting a scan body to the sleeve of the surgical guide instead of directly placing the implant. This helps to evaluate the accuracy of the surgical guide without errors in the placement of an implant.

Metabolic pathways modulate the neuronal toxicity associated with fragile X-associated tremor/ataxia syndrome.

Fragile X-associated tremor/ataxia syndrome (FXTAS) is an adult-onset neurodegenerative disorder that affects premutation carriers (55-200 CGG repeats) of the fragile X mental retardation 1 (FMR1) gene. Much remains unknown regarding the metabolic alterations associated with FXTAS, especially in the brain, and the most affected region, the cerebellum. Investigating the metabolic changes in FXTAS will aid in the identification of biomarkers as well as in understanding the pathogenesis of disease. To identify the metabolic alterations associated with FXTAS, we took advantage of our FXTAS mouse model that expresses 90 CGG repeats in cerebellar Purkinje neurons and exhibits the key phenotypic features of FXTAS. We performed untargeted global metabolic profiling of age-matched control and FXTAS mice cerebella at 16-20 weeks and 55 weeks. Out of 506 metabolites measured in cerebellum, we identified 186 metabolites that demonstrate significant perturbations due to the (CGG)90 repeat (P<0.05) and found that these differences increase dramatically with age. To identify key metabolic changes in FXTAS pathogenesis, we performed a genetic screen using a Drosophila model of FXTAS. Out of 28 genes that we tested in the fly, 8 genes showed significant enhanced neuronal toxicity associated with CGG repeats, such as Schlank (ceramide synthase), Sk2 (sphingosine kinase) and Ras (IMP dehydrogenase). By combining metabolic profiling with a Drosophila genetic screen to identify genetic modifiers of FXTAS, we demonstrate an effective method for functional validation of high-throughput metabolic data and show that sphingolipid and purine metabolism are significantly perturbed in FXTAS pathogenesis.

Rare variants in MYH15 modify amyotrophic lateral sclerosis risk.

Amyotrophic lateral sclerosis (ALS) is a fatal neurological disorder characterized by progressive muscular atrophy and respiratory failure. The G4C2 repeat expansion in the C9orf72 gene is the most prevalent genetic risk for ALS. Mutation carriers (C9ALS) display variability in phenotypes such as age-at-onset and duration, suggesting the existence of additional genetic factors. Here we introduce a three-step gene discovery strategy to identify genetic factors modifying the risk of both C9ALS and sporadic ALS (sALS) using limited samples. We first identified 135 candidate genetic modifiers of C9ALS using whole-genome sequencing (WGS) of extreme C9ALS cases diagnosed ~30 years apart. We then performed an unbiased genetic screen using a Drosophila model of the G4C2 repeat expansion with the genes identified from WGS analysis. This genetic screen identified the novel genetic interaction between G4C2 repeat-associated toxicity and 18 genetic factors, suggesting their potential association with C9ALS risk. We went on to test if 14 out of the 18 genes, those which were not known to be risk factors for ALS previously, are also associated with ALS risk in sALS cases. Gene-based-statistical analyses of targeted resequencing and WGS were performed. These analyses together reveal that rare variants in MYH15 represent a likely genetic risk factor for ALS. Furthermore, we show that MYH15 could modulate the toxicity of dipeptides produced from expanded G4C2 repeat. Our study presented here demonstrates the power of combining WGS with fly genetics to facilitate the discovery of fundamental genetic components of complex traits with a limited number of samples.

The CNK1 scaffold binds cytohesins and promotes insulin pathway signaling.

Protein scaffolds play an important role in signal transduction, regulating the localization of signaling components and mediating key protein interactions. Here, we report that the major binding partners of the Connector Enhancer of KSR 1 (CNK1) scaffold are members of the cytohesin family of Arf guanine nucleotide exchange factors, and that the CNK1/cytohesin interaction is critical for activation of the PI3K/AKT cascade downstream from insulin and insulin-like growth factor 1 (IGF-1) receptors. We identified a domain located in the C-terminal region of CNK1 that interacts constitutively with the coiled-coil domain of the cytohesins, and found that CNK1 facilitates the membrane recruitment of cytohesin-2 following insulin stimulation. Moreover, through protein depletion and rescue experiments, we found that the CNK1/cytohesin interaction promotes signaling from plasma membrane-bound Arf GTPases to the phosphatidylinositol 4-phosphate 5-kinases (PIP5Ks) to generate a PIP(2)-rich microenvironment that is critical for the membrane recruitment of insulin receptor substrate 1 (IRS1) and signal transmission to the PI3K/AKT cascade. These findings identify CNK1 as a new positive regulator of insulin signaling.

Comparison of digital intraoral scanner reproducibility and image trueness considering repetitive experience.

STATEMENT OF PROBLEM: Because the digital workflow can begin directly in the oral cavity, intraoral scanners are being adopted in dental treatments. However, studies of the relationship between the experience of the practitioner and the accuracy of impression data are needed. PURPOSE: The purpose of this clinical study was to investigate the effect of the experience curve on changes in trueness when a patient's complete dental arch is scanned. MATERIAL AND METHODS: Twenty dental hygienists with more than 3 years of experience in dental clinical practice (group 1 had 3 to 5 years; group 2 had >6 years) were recruited to learn to operate 2 intraoral scanner systems. All learners scanned the assigned patient's oral cavity 10 times during the experience sessions. Precision was calculated as the mean deviation among all superimposition combinations from the 10 scanned data sets of each learner [n=10C2=45]. Trueness was evaluated by superimposing the 10 consecutive intraoral scan data onto the impression scan data from each patient's rubber impression body (n=10). The acquired images were processed and analyzed using a 3-dimensional analysis software. For statistical analysis, the independent 2-sample t test and repeated measures ANOVA were performed (α=.05). RESULTS: The mean precision of the Trios scanner was greater than that of the iTero (Trios, 52.30 µm; iTero, 60.46 µm; P<.01). The iTero group showed an improvement in trueness upon repeated experience (P<.05), whereas the Trios group did not (P>.05). In the iTero group but not in the Trios group, the length of clinical experience influenced the change of trueness as a result of repeated experience (P<.05). In terms of the scanned region, the results for trueness were better for the maxillary arch than the mandibular arch with repeated scanning in the iTero group (P<.05). CONCLUSIONS: The single-image based system required repeated learning sessions for effective clinical application. The newer system offered better trueness and precision and was less likely to be influenced by the length of clinical career or the region being scanned.

Current utilization and influencing factors of complementary and alternative medicine among children with neuropsychiatric disease: a cross-sectional survey in Korea.

BACKGROUND: Complementary and alternative medicine (CAM) is widespread but has various utilization rates according to country and the condition of patients. Generally, CAM is more frequently used in diseases that have no clear treatment method in conventional medicine. Therefore, a high utilization rate of CAM can be assumed in pediatric neurological diseases, but few studies have investigated the utilization of CAM in children with neuropsychiatric diseases. In particular, studies regarding the current use of CAM are scarce. METHODS: We conducted a survey of the parents or caregivers of patients who visited the pediatric rehabilitation clinic, pediatric neurology clinic, or pediatric psychiatry clinic at one university hospital from April to July 2011. We analyzed the factors that affect the utilization of CAM and other rehabilitation therapies. RESULTS: Among the 578 patients recruited, 258 patients have ever received CAM (51.5%), and the current CAM utilization rate was 19.0% (110 patients). Two hundred patients (34.6%) were currently receiving only other rehabilitation therapies, and 268 patients (46.4%) were currently receiving no type of therapy. The rate of current CAM usage was significantly high in epilepsy patients. The ORs of 1-6-year-old and 7-12-year-old children compared with 13-19-year-old children were 3.14 (95 % CI 1.31-7.53) and 3.34 (95% CI 1.64-6.79), respectively, and the OR of the group with longer disease duration (≥48 months) compared with the group with shorter disease duration was 3.36 (95% CI 1.71-6.59). Only the age and disease duration showed statistically significant differences between the patients who were administered CAM and those who received other rehabilitation therapies (p < 0.0001). CONCLUSIONS: CAM is preferred by patients under 13 years of age compared with patients aged 13-19 years, whereas other rehabilitation therapies are preferred by patients aged 1-6 years, followed by those aged 6-12 years and then by those aged 13-19 years. The patient's age and disease duration are the major factors influencing CAM use. Future studies should specify particular diseases, rather than combining all types of neuropsychiatric diseases, and include the socio-economic status of the parents.

EphrinB1 interacts with CNK1 and promotes cell migration through c-Jun N-terminal kinase (JNK) activation.

The Eph receptors and their membrane-bound ligands, ephrins, play important roles in various biological processes such as cell adhesion and movement. The transmembrane ephrinBs transduce reverse signaling in a tyrosine phosphorylation-dependent or -independent, as well as PDZ-dependent manner. Here, we show that ephrinB1 interacts with Connector Enhancer of KSR1 (CNK1) in an EphB receptor-independent manner. In cultured cells, cotransfection of ephrinB1 with CNK1 increases JNK phosphorylation. EphrinB1/CNK1-mediated JNK activation is reduced by overexpression of dominant-negative RhoA. Overexpression of CNK1 alone is sufficient for activation of RhoA; however, both ephrinB1 and CNK1 are required for JNK phosphorylation. Co-immunoprecipitation data showed that ephrinB1 and CNK1 act as scaffold proteins that connect RhoA and JNK signaling components, such as p115RhoGEF and MKK4. Furthermore, adhesion to fibronectin or active Src overexpression increases ephrinB1/CNK1 binding, whereas blocking Src activity by a pharmacological inhibitor decreases not only ephrinB1/CNK1 binding, but also JNK activation. EphrinB1 overexpression increases cell motility, however, CNK1 depletion by siRNA abrogates ephrinB1-mediated cell migration and JNK activation. Moreover, Rho kinase inhibitor or JNK inhibitor treatment suppresses ephrinB1-mediated cell migration. Taken together, our findings suggest that CNK1 is required for ephrinB1-induced JNK activation and cell migration.

Appetitive learning requires the alpha1-like octopamine receptor OAMB in the Drosophila mushroom body neurons.

Associative learning is a fundamental form of behavioral plasticity. Octopamine plays central roles in various learning types in invertebrates; however, the target receptors and underlying mechanisms are poorly understood. Drosophila provides a powerful system to uncover the mechanisms for learning and memory. Here, we report that OAMB in the mushroom body neurons mediates the octopamine's signal for appetitive olfactory learning. The octopamine receptor OAMB has two isoforms (OAMB-K3 and OAMB-AS), differing in the third cytoplasmic loop and downstream sequence. The activation of each OAMB isoform increases intracellular Ca(2+) similar to the alpha1 adrenergic receptor, while OAMB-K3 additionally stimulates cAMP production. The oamb-null mutants showed severely impaired learning in appetitive olfactory conditioning that tests flies' capacity to learn and remember the odor associated with sugar reward. This deficit was also seen in the hypomorphic mutant with reduced OAMB expression in the mushroom bodies, the brain structure crucial for olfactory conditioning. Consistently, the oamb mutant's learning phenotype was fully rescued by conditional expression of either OAMB isoform in the mushroom body αß and γ neurons. These results indicate that the OAMB receptor is a key molecule mediating the octopamine's signal for appetitive olfactory learning and its functional site is the mushroom body αß and γ neurons. This study represents a critical step forward in understanding the cellular mechanism and neural circuit mediating reward learning and memory.

Effects of washing agents on the mechanical and biocompatibility properties of water-washable 3D printing crown and bridge resin.

Three-dimensional (3D) printing, otherwise known as additive manufacturing in a non-technical context, is becoming increasingly popular in the field of dentistry. As an essential step in the 3D printing process, postwashing with organic solvents can damage the printed resin polymer and possibly pose a risk to human health. The development of water-washable dental resins means that water can be used as a washing agent. However, the effects of washing agents and washing times on the mechanical and biocompatibility properties of water-washable resins remain unclear. This study investigated the impact of different washing agents (water, detergent, and alcohol) and washing time points (5, 10, 20, and 30 min) on the flexural strength, Vickers hardness, surface characterization, degree of conversion, biocompatibility, and monomer elution of 3D printed samples. Using water for long-term washing better preserved the mechanical properties, caused a smooth surface, and improved the degree of conversion, with 20 min of washing with water achieving the same biological performance as organic solvents. Water is an applicable agent option for washing the 3D printing water-washable temporary crown and bridge resin in the postwashing process. This advancement facilitates the development of other water-washable intraoral resins and the optimization of clinical standard washing guidelines.

Influence of hydrothermal aging on the shear bond strength of 3D printed denture-base resin to different relining materials.

OBJECTIVES: This study evaluated the repairability of three-dimensional printed (3DP) denture bases based on different conventional relining materials and aging. MATERIAL AND METHODS: The groups for surface characterization (surface-roughness and contact-angle measurements) were divided based on the denture base and surface treatment. Shear bond strength test and failure-mode analysis were conducted by a combination of three variables: denture base, relining materials, and hydrothermal aging (HA). The initial characterization involved quantifying the surface roughness (n = 10) and contact angle (n = 10) of denture base specimens with and without sandblasting (SB) treatment. Four relining materials (Kooliner [K], Vertex Self-Curing [V], Tokuyama Rebase II (Normal) [T], and Ufi Gel Hard [U]) were applied to 3DP, heat-cured (HC), and self-cured (SC) denture-base resin specimens. Shear bond strength (n = 15) and failure-mode analyses (n = 15) were performed before and after HA, along with evaluations of the fractured surfaces (n = 4). Statistical analyses were performed using a two-way analysis of variance (ANOVA) for surface characterization, and a three-way ANOVA was conducted for shear bond strength. RESULTS: The surface roughness peaked in HC groups and increased after SB. The 3DP group displayed significantly lower contact angles, which increased after treatment, similar to the surface roughness. The shear bond strength was significantly lower for 3DP and HC denture bases than for SC denture bases, and peaked for U at 10.65 ± 1.88 MPa (mean ± SD). HA decreased the shear bond strength relative to untreated samples. Furthermore, 3DP, HC, and SC mainly showed mixed or cohesive failures with V, T, and U. K, on the other hand, trended toward adhesive failures when bonded with HC and SC. CONCLUSION: This study has validated the repairability of 3DP dentures through relining them with common materials used in clinical practice. The repairability of the 3DP denture base was on par with that of conventional materials, but it decreased after aging. Notably, U, which had a postadhesive application, proved to be the most effective material for repairing 3DP dentures.

Three-axis load analysis of high-speed handpiece on dental training teeth and computer-aided design/computer-aided manufacturing blocks.

This study aimed to evaluate and compare the mechanical properties of dental training teeth with subtractive and additive computer-aided design/computer-aided manufacturing (CAD/CAM) materials used to fabricate dental simulation models. Therefore, the three-axis load generated during cutting movements, including drilling and milling performed using a dental handpiece, was measured and compared. The samples were cut vertically downward by 1.5 mm, horizontally by 6 mm, and vertically upward at a constant speed (1 mm/s), while the rotational speed of the bur was maintained at 200,000 rotations per minute. A three-axis load cell was used to measure the X-, Y-, and Z-axis loads on the specimen. The median value of the X-, Y-, and Z-axis measurements and the resultant load during the vertical-downward, horizontal, and vertical-upward movements were compared using a one-way analysis of variance and Tukey's post hoc test. For vertical downward movement, the drilling force of the dental training teeth was lower than that of Vita Enamic® and similar to that of Lava™ Ultimate. In contrast to subtractive CAD/CAM blocks, the drilling force of the dental training teeth was higher than that of 3D-printed resin blocks. Regarding horizontal movement, the milling force of dental training teeth was lower than that of Vita Enamic®. In contrast, the milling force of Nissin was similar to that of Lava™ Ultimate, while that of Frasaco was lower. Furthermore, compared to additive CAD/CAM blocks, the milling force of the dental training teeth was higher than that of 3D-printed resin blocks. Regarding vertical upward movement, the resultant loads of dental training teeth was lower than that of Vita Enamic®. Similarly, the resultant load of Nissin was similar to that of Lava™ Ultimate, while that of Frasaco was lower. Additionally, compared to additive CAD/CAM blocks, the resultant loads of the dental training teeth were similar to those of the 3D-printed resin blocks.

Effects of washing solution temperature on the biocompatibility and mechanical properties of 3D-Printed dental resin material.

The use of digital manufacturing, particularly additive manufacturing using three-dimensional (3D) printing, is expanding in the field of dentistry. 3D-printed resin appliances must undergo an essential process, post-washing, to remove residual monomers; however, the effect of the washing solution temperature on the biocompatibility and mechanical properties remains unclear. Therefore, we processed 3D-printed resin samples under different post-washing temperatures (without temperature control (N/T), 30 °C, 40 °C, and 50 °C) for different durations (5, 10, 15, 30, and 60 min) and evaluated the degree of conversion rate, cell viability, flexural strength, and Vickers hardness. Increasing the washing solution temperature significantly improved the degree of conversion rate and cell viability. Conversely, increasing the solution temperature and time decreased the flexural strength and microhardness. This study confirmed that the washing temperature and time influence the mechanical and biological properties of the 3D-printed resin. Washing 3D-printed resin at 30 °C for 30 min was most efficient to maintain optimal biocompatibility and minimize changes of mechanical properties.

Traditional East Asian Herbal Medicine for Post-Stroke Insomnia: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

Post-stroke insomnia (PSI) is a highly prevalent complication after stroke. Current evidence of psychotropic drug use for PSI management is scarce and indicates harmful adverse events (AEs). Traditional East Asian herbal medicine is a widely used traditional remedy for insomnia. However, so far, no study has systematically reviewed the efficacy and safety of traditional east asian herbal medicine (HM) for PSI. Therefore, we perform meta-analysis to evaluate the effectiveness and safety of HM for PSI. After a comprehensive electronic search of 15 databases, we review the randomized controlled trials (RCTs) of HM use as monotherapy for PSI. Our outcomes were the Pittsburgh sleep quality index and total effective rate. In total, 24 RCTs were conducted with 1942 participants. HM showed statistically significant benefits in sleep quality. It also appeared to be safer than psychotropic drugs in terms of AEs, except when the treatment period was two weeks. The methods used for RCTs were poor, and the quality of evidence assessed was graded "low" or "moderate." The findings of this review indicate that the use of HM as a monotherapy may have potential benefits in PSI treatment when administered as an alternative to conventional medications. However, considering the methodological quality of the included RCTs, we were uncertain of the clinical evidence. Further, well-designed RCTs are required to confirm these findings.

Effects of Post-Curing Light Intensity on the Mechanical Properties and Three-Dimensional Printing Accuracy of Interim Dental Material.

This study evaluated the effects of the light intensity of curing and the post-curing duration on the mechanical properties and accuracy of the interim dental material. After designing the specimen, 3D printing was performed, and the light intensity was divided into groups G20, G60, G80, and G120 (corresponding to 1.4−1.6, 2.2−3.0, 3.8−4.4, and 6.4−7.0 mW/cm2, respectively), with no post-curing or 5, 10, or 20 min of post-curing being performed. The flexural properties, Vickers microhardness, degree of conversion (DC), and 3D accuracy were then evaluated. The flexural properties and Vickers microhardness showed a sharp increase at the beginning of the post-curing and then tended to increase gradually as the light intensity and post-curing time increased (p < 0.001). On the other hand, there was no significant difference between groups in the accuracy analysis of a 3D-printed three-unit bridge. These results indicate that the light intensity of the post-curing equipment influences the final mechanical properties of 3D-printed resin and that post-curing can be made more efficient by optimizing the light intensity and post-curing time.