Purpose: This in vitro study investigated the effect of grooves, remaining tooth structure, and their combination on tooth fracture resistance of endodontically treated anterior teeth with cast dowel and cores. Materials and Methods: Sixty extracted maxillary anterior teeth of similar dimensions were endodontically treated and then randomly divided into three groups of 20 teeth each. The teeth in the first group were cut horizontally at the widest part of their anatomical crowns. Three hundred and sixty degree 1 and 2 mm axial walls for ferrule effect were provided for the teeth in the second and the third groups, respectively. Cast dowel and cores were fabricated for all teeth. Each group was then subdivided in two groups: one with no grooves and another with mesial and distal grooves. Hence, six groups were created as follows: (1) teeth with no remaining coronal tooth structure and no grooves (group A-control); (2) teeth with no remaining coronal tooth structure, with mesial and distal grooves (group B); (3) teeth with 1 mm remaining coronal tooth structure, with no grooves (group C); (4) teeth with 1 mm remaining coronal tooth structure, with mesial and distal grooves (group D); (5) teeth with 2 mm remaining coronal tooth structure and no grooves (group E); and (6) teeth with 2 mm remaining coronal tooth structure, with mesial and distal grooves (group F). Complete cast crowns were then fabricated for all teeth. A universal testing machine applied controlled loads to the teeth at a crosshead speed of 2.54 mm/min at an angle of 130• to the long axes of the teeth until failure occurred. The loads were applied 2 mm lower than the incisal edges of the specimens. Descriptive statistics, one-way ANOVA (α = 0.05) and Tukey's honestly significant difference (HSD) tests were used to determine the effect of failure loads among the tested groups (α = 0.05). Results: The mean failure loads were (N): group A (control), 151.21 ± 38.18; group B, 221.53 ± 107.03; group C, 295. 35 ± 81.92; group D, 270.20 ± 76.01; group E, 491.70 ± 180.36; group F, 432.67 ± 193.83. Group E presented the highest failure load and group A (control) the lowest. Conclusion:The inclusion of interproximal grooves on the cast dowel and cores of endodontically treated anterior teeth with 1-2 mm of remaining coronal tooth structure does not significantly lower the failure threshold.Endodontically treated anterior teeth do not require complete coverage restoration if they are not structurally compromised. Lovdahl and Nicholls 1 concluded that the intact endodontically treated central incisors were three times more resistant to fracture than teeth restored with dowels; however, tooth structure of endodontically treated teeth is frequently compromised due

Steroid 5-α reductase (5AR) is responsible for the reduction of steroids to 5-α reduced metabolites, such as the reduction of testosterone to 5-α dihydrotestosterone (DHT). A new adverse outcome pathway (AOP) for 5AR inhibition to reduce female reproduction in fish (AOP 289) is under development to clarify the antiestrogenic effects of 5AR inhibitors in female fish. A sensitive method for the DHT analysis using chemical derivatization and liquid chromatography–tandem mass spectrometry was developed. A cell-based 5AR inhibition assay that utilizes human cell lines, a transient overexpression system, and fish cell lines was developed. The measured IC50 values of two well-known 5AR inhibitors, finasteride and dutasteride, were comparable in the different systems. However, the IC50 of dutasteride in the fish cell lines was lower than that in the human cell lines. Finasteride showed a higher IC50 against the RTG-2 cell line. These results demonstrated that 5ARs inhibition could differ in terms of structural characteristics among species. The assay has high sensitivity and reproducibility and is suitable for the application in 5AR inhibition screening for various endocrine disruption chemicals (EDCs). Future studies will continue to evaluate the quantitative inhibition of 5AR by EDCs to compare the endocrine-disrupting pathway in different species.

No abstract

The disruption of thyroid hormones because of chemical exposure is a significant societal problem. Chemical evaluations of environmental and human health risks are conventionally based on animal experiments. However, owing to recent breakthroughs in biotechnology, the potential toxicity of chemicals can now be evaluated using 3D cell cultures. In this study, the interactive effects of thyroid‐friendly soft (TS) microspheres on thyroid cell aggregates are elucidated and their potential as a reliable toxicity assessment tool is evaluated. Using state‐of‐the‐art characterization methods coupled with cell‐based analysis and quadrupole time‐of‐flight mass spectrometry, it is shown that TS‐microsphere‐integrated thyroid cell aggregates exhibit improved thyroid function. Specifically, the responses of zebrafish embryos, which are used for thyroid toxicity analysis, and the TS‐microsphere‐integrated cell aggregates to methimazole (MMI), a known thyroid inhibitor, are compared. The results show that the thyroid hormone disruption response of the TS‐microsphere‐integrated thyroid cell aggregates to MMI is more sensitive compared with those of the zebrafish embryos and conventionally formed cell aggregates. This proof‐of‐concept approach can be used to control cellular function in the desired direction and hence evaluate thyroid function. Thus, the proposed TS‐microsphere‐integrated cell aggregates may yield new fundamental insights for advancing in vitro cell‐based research.

Mono(2-ethylhexyl)-phthalate (MEHP) is the primary metabolite of di(2-ethylhexyl)-phthalate (DEHP), which is widely used in industry as a plasticizer. Previous studies showed that both DEHP and MEHP have been found in not only human urine samples but also natural aquatic environments, and well documented as toxicants for reproduction and endocrine disruptors, However, the effects of MEHP exposure on aquatic organisms, including invertebrates such as Daphnia magna (D. magna), are still scarce. In the present study, the lipid alterations caused by MEHP in D. magna have been identified by analyzing lipid accumulation and nontarget metabolomics. Additionally, the expressions of reproduction were investigated. Ecotoxicologically relevant concentrations (1 to 2 mg/L) and exposure time ranges (24 h to 21 days) have been tested. MEHP resulted in no mortality to D. magna with all exposure conditions, but the number of lipid droplets increased after 96 h of exposure and enhanced reproduction of female adult daphnids was observed in the 21-day of exposure. Besides, we observed that MEHP enhanced lipid metabolism with the identification of 283 potential lipid metabolites, including glycerolipids, glycerophospholipids, and sphingolipids, following 48 h of exposure. MEHP treated group exhibited significantly higher expression of ecdysone receptor (EcR) and vitellogenin 2 (Vtg2) at 6 h and 24 h. Meanwhile at 48 h, EcR and Vtg2 were downregulated in 1 and 2 mg/L MEHP exposure. Our data show that the changes in EcR pathway with different exposure time could be associated with the lipid accumulation due to lipids increment subsequently to increased reproduction in MEHP-exposed D. magna.

No abstract

The SARS-CoV-2 pandemic requires a new therapeutic target for viral infection, and papain-like protease (Plpro) has been suggested as a druggable target. This in-vitro study was conducted to examine the drug metabolism of the GRL0617 and HY-17542, Plpro inhibitors. Metabolism of these inhibitors was studied to predict the pharmacokinetics in human liver microsomes. The hepatic cytochrome P450 (CYP) isoforms responsible for their metabolism were identified using recombinant enzymes. The drug–drug interaction potential mediated by cytochrome P450 inhibition was estimated. In human liver microsomes, the Plpro inhibitors had phase I and phase I + II metabolism with half-lives of 26.35 and 29.53 min, respectively. Hydroxylation (M1) and desaturation (-H2, M3) of the para-amino toluene side chain were the predominant reactions mediated with CYP3A4 and CYP3A5. CYP2D6 is responsible for the hydroxylation of the naphthalene side ring. GRL0617 inhibits major drug-metabolizing enzymes, including CYP2C9 and CYP3A4. HY-17542 is structural analog of GRL0617 and it is metabolized to GRL0617 through non-cytochrome P450 reactions in human liver microsomes without NADPH. Like GRL0617 and HY-17542 undergoes additional hepatic metabolism. The in-vitro hepatic metabolism of the Plpro inhibitors featured short half-lives; preclinical metabolism studies are needed to determine therapeutic doses for these inhibitors.

Mono(2-ethylhexyl)-phthalate (MEHP) is the primary metabolite of di(2-ethylhexyl)-phthalate (DEHP), which is widely used in industry as a plasticizer. Previous studies showed that both DEHP and MEHP have been found in not only human urine samples but also natural aquatic environments, and well documented as toxicants for reproduction and endocrine disruptors, However, the effects of MEHP exposure on aquatic organisms, including invertebrates such as Daphnia magna (D. magna), are still scarce. In the present study, the lipid alterations caused by MEHP in D. magna have been identi ed by analyzing lipid accumulation and nontarget metabolomics. Additionally, the expressions of reproduction were investigated. Ecotoxicologically relevant concentrations (1 to 2 mg/L) and exposure time ranges (24 h to 21 days) have been tested. MEHP resulted in no mortality to D. magna with all exposure conditions, but the number of lipid droplets increased after 96 h of exposure and enhanced reproduction of female adult daphnids was observed in the 21-day of exposure. Besides, we observed that MEHP enhanced lipid metabolism with the identi cation of 283 potential lipid metabolites, including glycerolipids, glycerophospholipids, and sphingolipids, following 48 h of exposure. MEHP treated group exhibited signi cantly higher expression of ecdysone receptor (EcR) and vitellogenin 2 (Vtg2) at 6 h and 24 h. Meanwhile at 48 h, EcR and Vtg2 were downregulated in 1 and 2 mg/L MEHP exposure. Our data show that the changes in EcR pathway with different exposure time could be associated with the lipid accumulation due to lipids increment subsequently to increased reproduction in MEHP-exposed D. magna.