Correction: Wang et al. Improve Dentin Bonding Performance Using a Hydrolytically Stable, Ether-Based Primer. J. Funct. Biomater. 2022, 13, 128.

Jirun Sun was not included as an author in the original publication [...].

Improve Dentin Bonding Performance Using a Hydrolytically Stable, Ether-Based Primer.

The objective of this study is to replace a traditional methacrylate-based primer (glycine, N-(2-hydroxy-3-(2-methyl-1-oxo-2-propenyl)propyl)-N-(4-methylphenyl) monosodium salt, NTG-GMA) with a hydrolytically stable ether-based primer (glycine, N-2-hydroxy-3-(4-vinylbenzyloxy)-propyl-N-(4-methylphenyl), monosodium salt, NTG-VBGE). The performance and durability of bonding composites to detin of two primers combined with methacrylate-based or ether-based adhesives were evaluated using shear bond strength (SBS) and micro-tensile bond strength (µTBS) combined with thermal cycling. The hydrolysis resistance of NTG-VBGE against hydrolysis was tested by challenging primed hydroxyapatite crystals with an esterase. The hydrophilicity of the primers and the resin spreading kinetics of adhesives on primed dentin were characterized by water contact angle measurements. The new primer NTG-VBGE was found to be compatible with both methacrylate-based adhesives and ether-based adhesives. The highest µTBS values were found in the test group of NTG-VBGE and ether-based adhesive, which was consistent with the resin spreading kinetics results. The more hydrophobic and hydrolytically stable primer/adhesive achieved improved dentin infiltration and bonding strength, suggesting significant potential for further developing dental restorative materials with extended service life.

Clinical evaluation of monolithic zirconia crowns: a failure analysis of clinically obtained cases from a 3.5-year study.

PURPOSE: The primary purpose of this study was to examine the clinical performance of monolithic zirconia single crowns in terms of short-term failure or complications. The secondary purpose was to detect the originating flaws of clinically failed monolithic zirconia crowns to find the causes of failure. METHODS: A short-term prospective cohort study based on record evaluation and clinical examination of patients treated with tooth-supported monolithic zirconia crowns was performed in the Department of Fixed Prosthodontics, Tohoku University Hospital, Japan. The crowns were prepared during the follow-up period from April 2014 to July 2018. The 3.5-year cumulative success and survival rates were set as primary endpoints. Fractures of the crown or fragments were inspected under a scanning electron microscope for descriptive fractography. RESULTS: During the study period, 40 monolithic zirconia crowns were placed. Four crowns experienced clinical complications, including: 1) fracture of the crown (two crowns), 2) abrasion of the crown (one crown), and 3) fracture of the antagonist tooth (one crown). The estimated Kaplan-Meier 3.5-year success and survival rates were 90.5% (95% confi dence interval [CI]: 73.1-97.1) and 92.8% (95% CI: 74.1-98.3), respectively. Fractography revealed that all fractures were initiated from the wear phase on the occlusal surface. CONCLUSIONS: The results of this study suggest that the molar application of monolithic zirconia crowns requires detailed attention to interocclusal clearance and the restoration of the antagonist tooth.

A thick frame decreases the fracture toughness of veneering ceramics used for zirconia-based all-ceramic restorations.

PURPOSE: To investigate the influence of firing condition and thickness of a yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) frame on the fracture toughness of veneering ceramics. METHODS: Layering and pressable ceramics without frames or on 0.8- and 1.2-mm-thick Y-TZP frames were fired with 5 schedules (normal and 4 customized conditions). The fracture toughness of the ceramics was measured at 0.5-2.5mm from the frame by the indentation fracture method. Crystal structures at the measurement points were evaluated by X-ray diffraction (XRD) and scanning electron microscopy (SEM). RESULTS: Under customized firing conditions, the fracture toughness of the layering ceramic significantly decreased, particularly near the frame. Use of a 1.2-mm-thick frame significantly decreased the fracture toughness of both layering and pressable ceramics compared to ceramics without the frame. The decreased fracture toughness only occurred close (0.5 and 1.0mm) to the frame. XRD analysis showed identical diffraction patterns between points near and far from the frame, suggesting that the decreased fracture toughness was not due to crystal defects or impurities. SEM revealed many microcracks and large crystals in layering ceramic near the 1.2-mm-thick frame, possibly resulting from the thermal properties of the zirconia frame, such as large heat capacity and low thermal conductivity. CONCLUSIONS: Modification of the firing conditions from the manufacturer's instructions and inclusion of a thick Y-TZP frame decreased the fracture toughness of veneering ceramics at the interface, suggesting that a thick frame would pose a potent fracture risk in veneering ceramics used for zirconia-based all-ceramic restorations.

A possible risk of CAD/CAM-produced composite resin premolar crowns on a removable partial denture abutment tooth: a 3-year retrospective cohort study.

PURPOSE: To evaluate the early performance of computer-aided design/computer-aided manufacturing (CAD/CAM)-produced composite resin crown (CAD/CAM composite crown) treatment on premolars, specifically, placement on a removable partial denture (RPD) abutment tooth, and the distalmost tooth in the dental arch, as possible clinical risk factors. METHODS: A retrospective cohort study (April 2014 to July 2017) was performed utilizing the clinical records of patients who received a premolar CAD/CAM composite crown treatment. The variables of time of treatment for (1) successful crowns (complication event-free) and (2) surviving crowns (clinically functional including re-luted) were estimated using Kaplan-Meier analysis. Survival distributions regarding "RPD abutment tooth" and "distalmost tooth" were analyzed with the log-rank test. Multilevel survival analyses were used to identify hazard ratios and associated risk factors. RESULTS: Five hundred and forty-seven crowns were evaluated (mean follow-up time 1.3±0.9years) in 327 patients. A total of 87 crowns had at least one complication, with loss of crown retention being the most common (70 crowns). Estimated success and survival rates at 3 years were 71.7% and 96.4%, respectively. The risk of complications was significantly higher for an RPD abutment tooth than for a non-RPD abutment tooth. There was no significant difference between the distalmost tooth and non-distalmost tooth in the dental arch. CONCLUSIONS: The demonstrated complication rate for CAD/CAM composite crowns placed on premolars was 15.9% over a period of up to 3 years. There was a substantial risk of complications with placement of such a crown on an RPD abutment tooth.

Effect of finish line design on stress distribution in bilayer and monolithic zirconia crowns: a three-dimensional finite element analysis study.

This study evaluated the influence of different finish line designs and abutment materials on the stress distribution of bilayer and monolithic zirconia crowns using three-dimensional finite element analysis (FEA). Three-dimensional models of two types of zirconia premolars - a yttria-stabilized zirconia framework with veneering ceramic and a monolithic zirconia ceramic - were used in the analysis. Cylindrical models with the finish line design of the crown abutments were prepared with three types of margin curvature radius (CR): CR = 0 (CR0; shoulder margin), CR = 0.5 (CR0.5; rounded shoulder margin), and CR = 1.0 (CR1.0; deep chamfer margin). Two abutment materials (dentin and brass) were analyzed. In the FEA model, 1 N was loaded perpendicular to the occlusal surface at the center of the crown, and linear static analysis was performed. For all crowns, stress was localized to the occlusal loading area as well as to the axial walls of the proximal region. The lowest maximum principal stress values were observed when the dentin abutment with CR0.5 was used under a monolithic zirconia crown. These results suggest that the rounded shoulder margin and deep chamfer margin, in combination with a monolithic zirconia crown, potentially have optimal geometry to minimize occlusal stress.

Clinical evaluation of zirconia-based all-ceramic single crowns: an up to 12-year retrospective cohort study.

OBJECTIVES: This study aims to investigate the incidence of clinical complications with tooth-supported zirconia-based all-ceramic single crowns and identify pertinent risk parameters. MATERIALS AND METHODS: A retrospective cohort study (May 2004 to April 2016) utilizing clinical records of patients receiving yttrium-oxide-partially stabilized zirconia (Y-TZP)-based all-ceramic crowns placed at Tohoku University Hospital was performed. The length of time of treatment success (complication event-free) and restoration survival (including minor complication events and remaining clinically functional) were estimated using Kaplan-Meier analysis. Multilevel survival analysis was used to identify risk factors. RESULTS: One hundred thirty-seven crowns were evaluated (mean follow-up time, 7.0 years). A total of 21 crowns experienced at least one complication with fracture of veneering ceramic being the most common (16 crowns). Estimated success and survival rates at 5 years (96.9 and 98.5%, respectively) decreased at 10 years to 62.1 and 67.2%, respectively. The risk of complications was significantly higher for molar crowns compared to anterior crowns (p < 0.01). A significant association of complications with metal antagonist restorations was shown by univariate analysis (p < 0.01). CONCLUSIONS: Given the study limitations, Y-TZP single crowns placed on anterior teeth demonstrated encouraging clinical results over a period of up to 10 years. However, there is a substantial risk of complications with posterior teeth within 10 years of restoration placement. CLINICAL RELEVANCE: Treatment with zirconia-based all-ceramic crowns for molar teeth with metal antagonist occlusion should be undertaken with caution.

Three-dimensional finite element analysis of zirconia all-ceramic cantilevered fixed partial dentures with different framework designs.

The purpose of this study were: to perform stress analyses using three-dimensional finite element analysis methods; to analyze the mechanical stress of different framework designs; and to investigate framework designs that will provide for the long-term stability of both cantilevered fixed partial dentures (FPDs) and abutment teeth. An analysis model was prepared for three units of cantilevered FPDs that assume a missing mandibular first molar. Four types of framework design (Design 1, basic type; Design 2, framework width expanded buccolingually by 2 mm; Design 3, framework height expanded by 0.5 mm to the occlusal surface side from the end abutment to the connector area; and Design 4, a combination of Designs 2 and 3) were created. Two types of framework material (yttrium-oxide partially stabilized zirconia and a high precious noble metal gold alloy) and two types of abutment material (dentin and brass) were used. In the framework designs, Design 1 exhibited the highest maximum principal stress value for both zirconia and gold alloy. In the abutment tooth, Design 3 exhibited the highest maximum principal stress value for all abutment teeth. In the present study, Design 4 (the design with expanded framework height and framework width) could contribute to preventing the concentration of stress and protecting abutment teeth.

Sudden Death Caused by Anomalous Origin of the Coronary Artery During Exercise.

Anomalous origin of the coronary artery (AOCA) is a rare, but important cause of sudden cardiac death among young athletes. Nine autopsy cases (8 male, 1 female; mean age, 17.9 years; age range, 11-31 years) of sudden death during or just after exercise caused by AOCA were reviewed. The exercises performed at the time of death were running (4 cases), soccer (2 cases), and baseball, swimming and kendo (Japanese swordsmanship) (1 case each). In 6 cases, the left coronary artery arose from the right sinus of Valsalva, and in 3, the right coronary artery from the left sinus. The coronary arteries passed between the pulmonary artery and the aorta with an acute angle takeoff from the orifice. Three cases had cardiovascular manifestations prior to death. In cases with cardiovascular manifestations, novel imaging methods should be considered to prevent sudden death.

An autopsy case of fatal repellent air freshener poisoning.

We describe a first fatal case of repellent air freshener ingestion. A 79-year-old Japanese man with Alzheimer-type senile dementia orally ingested repellent air freshener containing three surfactants: polyoxyethylene 9-lauryl ether, polyoxyethylene (40) hydrogenated castor oil, and lauric acid amidopropyl amine oxide (weight ratio of 1.3%). About 1h after the collapse, he was in cardiopulmonary arrest and subsequently died 10h after his arrival. The forensic autopsy performed 5.5h after death revealed the 380ml of stomach contents with a strong mint perfume identical to that of the repellent air freshener and the findings of acute death. Toxicologically, 9.1µg/ml and 558.2µg/ml of polyoxyethylene 9-lauryl ether were detected from the serum and stomach contents taken at autopsy. Generally, ingestion of anionic or non-ionic surfactants have been considered as safe. However, because the patient suffered from cardiac insufficiency with a low dose of repellent air freshener ingestion, medical staff members must evaluate the elderly patient for cardiac and circulatory problems regardless of the ingested dose. Not only medical and nursing staff members, but also families who are obliged to care for elderly persons must be vigilant to prevent accidental ingestion of toxic substances generally used in the household.

A novel application of infrared imaging for the diagnosis of death by fire.

This paper demonstrates a novel and simple technique for the diagnosis of death by fire using a digital infrared (IR) camera system. At autopsy, the inhalation of soot by a fire victim is a definite indicator proving a vital reaction during the fire. However, there can be difficulties in confirming the presence of soot because of the relative lack of soot or the similarity in color between soot and surrounding tissues. To evaluate the effectiveness of IR imaging at autopsy, we acquired internal images of the respiratory and gastrointestinal system by both an ordinary color and IR cameras. Using our method, the inhaled soot is visible as black particles or deposits, while the blood is transparent and the surrounding tissue is whitened. This technique enables the detection of normally undiscriminating soot in an ordinary color image. This is the first report suggesting the usefulness of IR imaging in forensic autopsy for the diagnosis of death by fire.

Simultaneous identification of the enantiomers and diastereomers of N,O-di-trifluoroacetylated ephedrine and norephedrine in blood plasma using chiral capillary gas chromatography-mass spectrometry with selected ion monitoring.

A method for identifying the enantiomers of N,O-di-trifluoroacetylated ephedrine (EP) and norephedrine (NE) and the enantiomers of pseudoephedrine (PEP) and pseudonorephedrine (PNE) in plasma was developed using chiral capillary gas chromatography-mass spectrometry (GC-MS) with selected ion monitoring (SIM). N,O-Di-trifluoroacethyl (TFA) derivatization was accomplished in a dried hydrochloride extract of plasma (minimum quantity of 0.2 mL). An SIM GC-MS method with a ß-cyclodextrin chiral capillary column allowed the successful and simultaneous detection of each TFA-derivatized stereoisomer of EP, NE, PEP, PNE, and an internal standard (IS; S-(+)-ethylamphetamine). Each TFA-drivatized stereoisomer was identified using four mass fragment ions (m/z 140, 154, 168, and 230). The TFA-derivatized stereoisomers of EP, NE, PEP, PNE, and IS were separated completely and were detected with sufficient sensitivity. The assay allowed the stereoisomers to be determined in a linear range of 12.5-1250 ng/mL for the EP stereoisomers and a linear range of 5-1250 ng/mL for the PEP, NE, and PNE stereoisomers. The detection limits were 7.5 ng/mL for the EP stereoisomers and 2.5 ng/mL for the PEP, NE, and PNE stereoisomers. The intra- and interday precisions were less than 5.9% and 8.2%, respectively. This chiral capillary SIM GC-MS method was sufficiently effective in the analysis of plasma from users of over-the-counter cold medicines and was also fully applicable to the plasma analysis of guinea pigs following their treatment with racemic EP.

Differential gene expression of multiple chondroitin sulfate modification enzymes among neural stem cells, neurons and astrocytes.

Chondroitin sulfate/dermatan sulfate (CS/DS) polysaccharides have been reported to play a crucial role in the proliferation and maintenance of neural stem cells (NSCs). However, little is known about the structural changes and functional role of CS/DS chains in the differentiation of NSCs. Western blots of NSCs, neurons and astrocytes in culture, with three CS-polysaccharide antibodies of different specificities, revealed marked differences in CS structure among the three cell types. To confirm this finding, we measured gene expression levels of CS sulfotransferases and C5-epimerase in these cell types, as these are responsible for producing the high structural diversity of CS/DS. Expressions of chondroitin 4-O-sulfotransferase, chondroitin 6-O-sulfotransferase, and N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase mRNAs were low in cultures of differentiated neural cells, such as neurons and astrocytes, in comparison to NSCs. In contrast, expressions of uronyl 2-O-sulfotransferase and C5-epimerase mRNAs were higher in the differentiated neural cells than NSCs. Thus, we first provide evidence to support the hypothesis that CS/DS undergoes structural changes during NSC differentiation. The structural changes in CS/DS may be implicated in the regulation of NSC differentiation through interactions with growth/neurotrophic factors and cytokines.

Yip1A regulates the COPI-independent retrograde transport from the Golgi complex to the ER.

Yip1A, a mammalian homologue of yeast Yip1p, is a multi-spanning membrane protein that is considered to be involved in transport between the endoplasmic reticulum (ER) and the Golgi. However, the precise role of Yip1A in mammalian cells remains unclear. We show here that endogenous Yip1A is localized to the ER-Golgi intermediate compartment (ERGIC). Knockdown of Yip1A by RNAi did not induce morphological changes in the Golgi, ER, or ERGIC. By analyzing a number of intracellular transport pathways, we found that Yip1A knockdown delayed the transport of Shiga toxin from the Golgi to the ER, but did not affect the anterograde transport of VSVGts045. We also found that a recombinant protein that corresponded to the N-terminal domain of Yip1A inhibited the COPI-independent retrograde transport of GFP-tagged galactosyltransferase, GT-GFP, but not the COPI-dependent retrograde transport of p58/ERGIC53. Furthermore, we found that Yip1A knockdown resulted in the dissociation of Rab6 from the membranes. These results suggested that Yip1A has a role in COPI-independent retrograde transport from the Golgi to the ER and regulates the membrane recruitment of Rab6.

Expression and identification of a new splice variant of neuroglycan C, a transmembrane chondroitin sulfate proteoglycan, in the human brain.

Neuroglycan C (NGC) is a transmembrane chondroitin sulfate proteoglycan with an EGF module. We studied the expression of NGC in the human brain, mainly in the hippocampus, and confirmed some observations by conducting experiments using rat brain. In humans, NGC mRNA was expressed exclusively in the brain, especially in the immature brain. The telencephalon, including the hippocampus and neocortex, showed strong mRNA expression. NGC was immunolocalized to neuropils in the hippocampus and neocortex of the adult rat. RT-PCR experiments showed that four splice variants (NGC-I, -II, -III, and -IV) were expressed in the adult human hippocampus. By Western blotting, the expression as proteins of all splice variants except NGC-II was confirmed in the adult rat hippocampus. NGC-IV, which was first found in the present study, had the shortest cytoplasmic domain among the four variants. NGC-IV mRNA was expressed by neurons, but not by astrocytes, in culture prepared from the fetal rat hippocampus, suggesting that NGC-IV plays a role specific to neurons. In addition, the human NGC gene, which is registered as CSPG5, comprised six exons and was approximately 19 kb in size. In exon 2, a single nucleotide polymorphism resulting in Val188Gly in the NGC ectodomain was observed.

Glycosylation site for chondroitin sulfate on the neural part-time proteoglycan, neuroglycan C.

Neuroglycan C (NGC) is a membrane-spanning chondroitin sulfate (CS) proteoglycan that is expressed predominantly in the central nervous system (CNS). NGC dramatically changed its structure from a proteoglycan to a nonproteoglycan form with cerebellar development, whereas a small portion of NGC molecules existed in a nonproteoglycan form in the other areas of the mature CNS, suggesting that the CS glycosylation of NGC is developmentally regulated in the whole CNS. As primary cultured neurons and astrocytes from cerebral cortices expressed NGC in a proteoglycan form and in a nonproteoglycan form, respectively, CS glycosylation seems to be regulated differently depending on cell type. To investigate the glycosylation process, cell lines expressing a proteoglycan form of NGC would be favorable experimental models. When a mouse NGC cDNA was transfected into COS 1, PC12D, and Neuro 2a cells, only Neuro 2a cells, a mouse neuroblastoma cell line, expressed NGC bearing CS chains. In PC12D cells, although three intrinsic CS proteoglycans were detected, exogenously expressed NGC did not bear any short CS chains just like NGC in the mature cerebellum. This suggests that the addition of CS chains to the NGC core protein is regulated in a manner different from that of other CS proteoglycans. As the first step in investigating the CS glycosylation mechanism using Neuro 2a cells, we determined the CS attachment site as Ser-123 on the NGC core protein by site-directed mutagenesis. The CS glycosylation was not necessary for intracellular trafficking of NGC to the cell surface at least in Neuro 2a cells.

Cdc2 kinase-dependent disassembly of endoplasmic reticulum (ER) exit sites inhibits ER-to-Golgi vesicular transport during mitosis.

We observed the disassembly of endoplasmic reticulum (ER) exit sites (ERES) by confocal microscopy during mitosis in Chinese hamster ovary (CHO) cells by using Yip1A fused to green fluorescence protein (GFP) as a transmembrane marker of ERES. Photobleaching experiments revealed that Yip1A-GFP, which was restricted to the ERES during interphase, diffused throughout the ER network during mitosis. Next, we reconstituted mitotic disassembly of Yip1A-GFP-labeled ERES in streptolysin O-permeabilized CHO cells by using mitotic L5178Y cytosol. Using the ERES disassembly assay and the anterograde transport assay of GFP-tagged VSVGts045, we demonstrated that the phosphorylation of p47 by Cdc2 kinase regulates the disassembly of ERES and results in the specific inhibition of ER-to-Golgi transport during mitosis.

Phosphorylation of neuroglycan C, a brain-specific transmembrane chondroitin sulfate proteoglycan, and its localization in the lipid rafts.

Neuroglycan C (NGC) is a brain-specific transmembrane chondroitin sulfate proteoglycan. In the present study, we examined whether NGC could be phosphorylated in neural cells. On metabolic labeling of cultured cerebral cortical cells from the rat fetus with (32)P(i), serine residues in NGC were radiolabeled. Some NGC became detectable in the raft fraction from the rat cerebrum, a signaling microdomain of the plasma membrane, with cerebral development. NGC from the non-raft fraction, not the raft fraction, could be phosphorylated by an in vitro kinase reaction. The phosphorylation of NGC was inhibited by adding to the reaction mixture a recombinant peptide representing the ectodomain of NGC, but not by adding a peptide representing its cytoplasmic domain. NGC could be labeled by an in vitro kinase reaction using [gamma-(32)P]GTP as well as [gamma-(32)P]ATP, and this kinase activity was partially inhibited by 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazole, a selective inhibitor of casein kinase II. In addition to the intracellular phosphorylation, NGC was also phosphorylated at the cell surface by an ectoprotein kinase. This is the first report to demonstrate that NGC can be phosphorylated both intracellularly and pericellularly, and our findings suggest that a kinase with a specificity similar to that of casein kinase II is responsible for the NGC ectodomain phosphorylation.