• Molecules and Cells
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• v.42 no.1
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• pp.17-27
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• 2019

Ubiquitin-specific protease 44 (USP44) has been implicated in tumor progression and metastasis across various tumors. However, the function of USP44 in prostate cancers and regulatory mechanism of histone-modifying enzymes by USP44 in tumors is not well-understood. Here, we found that enhancer of zeste homolog 2 (EZH2), a histone H3 lysine 27 methyltransferase, is regulated by USP44. We showed that EZH2 is a novel target of USP44 and that the protein stability of EZH2 is upregulated by USP44-mediated deubiquitination. In USP44 knockdown prostate cancer cells, the EZH2 protein level and its gene silencing activity were decreased. Furthermore, USP44 knockdown inhibited the tumorigenic characteristics and cancer stem cell-like behaviors of prostate cancer cells. Inhibition of tumorigenesis caused by USP44 knockdown was recovered by ectopic introduction of EZH2. Additionally, USP44 regulates the protein stability of oncogenic EZH2 mutants. Taken together, our results suggest that USP44 promotes the tumorigenesis of prostate cancer cells partly by stabilizing EZH2 and that USP44 is a viable therapeutic target for treating EZH2-dependent cancers.

• BMB Reports
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• v.55 no.11
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• pp.553-558
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• 2022

Hepatocellular carcinoma (HCC) is dangerous cancer that often evades early detection because it is asymptomatic and an effective detection method is lacking. For people with chronic liver inflammation who are at high risk of developing HCC, a sensitive detection method for HCC is needed. In a meta-analysis of The Cancer Genome Atlas pan-cancer methylation database, we identified a CpG island in the USP44 promoter that is methylated specifically in HCC. We developed methylation-sensitive high-resolution melting (MS-HRM) analysis to measure the methylation levels of the USP promoter in cell-free DNA isolated from patients. Our MS-HRM assay correctly identified 40% of patients with early-stage HCC, whereas the α-fetoprotein test, which is currently used to detect HCC, correctly identified only 25% of early-stage HCC patients. These results demonstrate that USP44 MS-HRM analysis is suitable for HCC surveillance.

• Food Science of Animal Resources
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• v.44 no.2
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• pp.483-497
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• 2024

This study investigated the effects of ultra-ground seaweed powders (USP) on the physicochemical properties (proximate composition, mineral contents, pH, color, shear force, sensory evaluation, electronic nose, and electronic tongue) of restructured black goat jerky. Restructured black goat jerky was prepared using three different treatments, i.e., 3% (w/w) each of ultra-ground sea tangle (ST; Undaria pinnatifida), sea mustard (SM; Saccharina japonica), and sea string (SS; Gracilaria verrucosa) powders. Moisture and ash contents were significantly higher in the USP-treated group than in the control (p<0.05). Potassium, calcium, and zinc contents were significantly higher in the SM than in the other USP-treated groups (p<0.05). In contrast, pH values were significantly higher in the ST and SM than in the control and SS (p<0.05). CIE L^*, CIE a^*, CIE b^*, and shear force were significantly lower in the USP-treated groups than in the control (p<0.05). Sensory evaluation revealed no significant difference in taste, texture, seaweed-like odor, and goaty flavor (p<0.05). Principal component analysis (PCA) and peak graph analysis of the electronic nose showed that the SS differed the most from the control compared with the other USP-treated groups, owing to the seaweed odor of ultra-ground SS powder. The PCA and ranking analysis of the electronic tongue showed that the umami taste of the SM was higher than that of the control and other USP-treated groups. Therefore, the potassium, calcium, zinc contents, and umami taste of reconstituted black goat jerky were significantly higher in the SM than in the control and other USP-treated groups.

• Analytical Science and Technology
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• v.33 no.6
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• pp.252-261
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• 2020

Currently, the potentiometric titration and the high pressure liquid chromatography (HPLC) method were utilized in Korean Pharmacopoeia XII (KP XII) as well as other pharmacopoeias (USP, EP, BP) for determination of loperamide hydrochloride in raw materials and capsules, respectively. The research objective is to overcome the remaining drawbacks from current methods such as solubility of mobile phase (KP XII), less scientific approach (USP 43) or using paired-ion chromatography reagent which shows some limitations (BP2017 and other formulation monographs). The proposed method was optimized by Design of Experiment (DoE) tool to obtain the satisfied method for determination of loperamide hydrochloride. The optimal condition was performed on the common C18 column (150 mm × 4.6 mm; 5 ㎛) using isocratic elution with the mobile phase containing 40 mM of potassium phosphate monobasic (pH 3.0) and acetonitrile (56:44), at a flow rate of 0.7 mL/min. The optimized method was validated and met the requirements of the International Conference on Harmonization. The developed method was applied to determine loperamide hydrochloride in capsules and can be used to update the current monograph in KP XII.

• Molecules and Cells
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• v.44 no.8
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• pp.557-568
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• 2021

Global knockout of the BK channel has been proven to affect bone formation; however, whether it directly affects osteoblast differentiation and the mechanism are elusive. In the current study, we further investigated the role of BK channels in bone development and explored whether BK channels impacted the differentiation and proliferation of osteoblasts via the canonical Wnt signaling pathway. Our findings demonstrated that knockout of Kcnma1 disrupted the osteogenesis of osteoblasts and inhibited the stabilization of β-catenin. Western blot analysis showed that the protein levels of Axin1 and USP7 increased when Kcnma1 was deficient. Together, this study confirmed that BK ablation decreased bone mass via the Wnt/β-catenin signaling pathway. Our findings also showed that USP7 might have the ability to stabilize the activity of Axin1, which would increase the degradation of β-catenin in osteoblasts.

• Journal of the Korean Ceramic Society
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• v.44 no.12
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• pp.690-695
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• 2007

Scandia stabilized zirconia (ScSZ) is adapted for electrolyte material of solid oxide fuel cell (SOFC) because of its high ionic conductivity and chemical stability. ScMnSZ1 powder having a composition of $((ZrO_2)_{0.89}(Sc_2O_3)_{0.1}(MnO_2)_{0.01})$ is synthesized by ultrasonic spray pyrolysis (USP) method. Porous ScMnSZ1 powder is obtained by using a pore forming agent. Microstructure and morphology, particle size distribution of porous powder synthesized with 3wt% pore forming agent are investigated. Sintered ScMnSZ1 sample with ground fine powder are also investigated their microstructure and electrical conductivity. The electrical conductivity of sintered ScMnSZ1 samples with ground fine powder was 0.082 S/cm, 0.127 S/cm and 0.249 S/cm at $750^{\circ}C$, $800^{\circ}C$ and $900^{\circ}C$, respectively.

• Journal of Pharmaceutical Investigation
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• v.41 no.4
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• pp.239-247
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• 2011

Simvastatin is an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, which catalyzes the conversion of HMG-CoA to mevalonate, an early and rate-limiting step in the biosynthesis of cholesterol. Simvastatin has good permeability, but it also has low solubility (BCS class II), which reduces its bioavailability. To overcome this problem, a solid dispersion is formed using a spray-dryer with polymeric material carrier to potentially enhance the dissolution rate and extend drug absorption. As carriers for solid dispersion, Gelucire$^{(R)}$44/14 and Gelucire$^{(R)}$ 50/13 are semisolid excipients that greatly improve the bioavailability of poorly-soluble drugs. To avoid any toxic effects of an organic solvent, we used aqueous medium to melt Tween$^{(R)}$ 80 and distilled water. The structural behaviors of the raw materials and the solid dispersion were analyzed by differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD) and scanning electron microscopy (SEM). The DSC and PXRD data indicated that the crystalline structure of simvastatin was transformed to an amorphous structure through solid dispersion. Then, solid dispersion-based tablets containing 20 mg simvastatin were prepared with excipients. Dissolution tests were performed in distilled water and artificial intestinal fluid using the USP paddle II method. Compared with that of the commercial tablet (Zocor$^{(R)}$ 20 mg), the release of simvastatin from solid dispersion based-tablet was more efficient. Although the stability study is not complete, this solid dispersion system is expected to deliver poorly water-soluble drugs with enhanced bioavailability and less toxicity.

• Restorative Dentistry and Endodontics
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• v.47 no.4
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• pp.39.1-39.12
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• 2022

Objectives: This study investigated the prevalence and characteristics of external cervical resorption (ECR) regarding sex, age, tooth, stages of progression, and portal of entry, using cone-beam computed tomography (CBCT) scans. Materials and Methods: CBCT scans of 1,313 patients from a Brazilian subpopulation comprising 883 female and 430 male patients (mean age, 55.2 years), acquired using a PreXion 3D CBCT unit, were evaluated. All permanent teeth included in the scans were evaluated for the presence of ECR according to the 3-dimensional classification and the portal of entry. The association between the presence of ECR and the factors studied was assessed using the χ² test. Intra-observer agreement was analyzed with the kappa test (α = 0.05). Results: In total, 6,240 teeth were analyzed, of which 84 (1.35%) were affected by ECR. A significant association was found between the presence of ECR and sex, with a higher prevalence in male patients (p = 0.002). The most frequently affected teeth were the mandibular and maxillary central incisors. The most common height was the mid-third of the root. For the portal of entry, 44% of cases were on the proximal surfaces, 40.5% on the lingual/palatal surface and 15.5% on the buccal surface. Intra-observer agreement was excellent. Conclusions: The prevalence of ECR was 1.35%, with a higher prevalence in male patients and a wide age distribution. The mandibular and maxillary central incisors were the most commonly affected teeth, and cases of ECR most frequently showed a height into the mid-third of the root and proximal entry.

• Molecules and Cells
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• v.44 no.3
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• pp.146-159
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• 2021

DNA methylation, and consequent down-regulation, of tumour suppressor genes occurs in response to epigenetic stimuli during cancer development. Similarly, human oncoviruses, including human papillomavirus (HPV), up-regulate and augment DNA methyltransferase (DNMT) and histone deacetylase (HDAC) activities, thereby decreasing tumour suppressor genes (TSGs) expression. Ubiquitin-like containing PHD and RING finger domain 1 (UHRF1), an epigenetic regulator of DNA methylation, is overexpressed in HPV-induced cervical cancers. Here, we investigated the role of UHRF1 in cervical cancer by knocking down its expression in HeLa cells using lentiviral-encoded short hairpin (sh)RNA and performing cDNA microarrays. We detected significantly elevated expression of thioredoxin-interacting protein (TXNIP), a known TSG, in UHRF1-knockdown cells, and this gene is hypermethylated in cervical cancer tissue and cell lines, as indicated by whole-genome methylation analysis. Up-regulation of UHRF1 and decreased TXNIP were further detected in cervical cancer by western blot and immunohistochemistry and confirmed by Oncomine database analysis. Using chromatin immunoprecipitation, we identified the inverted CCAAT domain-containing UHRF1-binding site in the TXNIP promoter and demonstrated UHRF1 knockdown decreases UHRF1 promoter binding and enhances TXNIP expression through demethylation of this region. TXNIP promoter CpG methylation was further confirmed in cervical cancer tissue by pyrosequencing and methylation-specific polymerase chain reaction. Critically, down-regulation of UHRF1 by siRNA or UHRF1 antagonist (thymoquinone) induces cell cycle arrest and apoptosis, and ubiquitin-specific protease 7 (USP7), which stabilises and promotes UHRF1 function, is increased by HPV viral protein E6/E7 overexpression. These results indicate HPV might induce carcinogenesis through UHRF1-mediated TXNIP promoter methylation, thus suggesting a possible link between CpG methylation and cervical cancer.