• Molecules and Cells
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• 제46권7호
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• pp.441-450
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• 2023

β-Catenin (Ctnnb1) has been shown to play critical roles in the development and maintenance of epithelial cells, including the retinal pigment epithelium (RPE). Ctnnb1 is not only a component of intercellular junctions in the epithelium, it also functions as a transcriptional regulator in the Wnt signaling pathway. To identify which of its functional modalities is critically involved in mouse RPE development and maintenance, we varied Ctnnb1 gene content and activity in mouse RPE lineage cells and tested their impacts on mouse eye development. We found that a Ctnnb1 double mutant (Ctnnb1^dm), which exhibits impaired transcriptional activity, could not replace Ctnnb1 in the RPE, whereas Ctnnb1^Y654E, which has reduced affinity for the junctions, could do so. Expression of the constitutively active Ctnnb1^∆ex3 mutant also suppressed the development of RPE, instead facilitating a ciliary cell fate. However, the post-mitotic or mature RPE was insensitive to the loss, inactivation, or constitutive activation of Ctnnb1. Collectively, our results suggest that Ctnnb1 should be maintained within an optimal range to specify RPE through transcriptional regulation of Wnt target genes in the optic neuroepithelium.

• Molecules and Cells
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• 제46권8호
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• pp.476-485
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• 2023

Gastric cancer stem-like cells (GCSCs) possess stem cell properties, such as self-renewal and tumorigenicity, which are known to induce high chemoresistance and metastasis. These characteristics of GCSCs are further enhanced by autophagy, worsening the prognosis of patients. Currently, the mechanisms involved in the induction of stemness in GCSCs during autophagy remain unclear. In this study, we compared the cellular responses of GCSCs with those of gastric cancer intestinal cells (GCICs) whose stemness is not induced by autophagy. In response to glucose starvation, the levels of β-catenin and stemness-related genes were upregulated in GCSCs, while the levels of β-catenin declined in GCICs. The pattern of deubiquitinase ubiquitin C-terminal hydrolase-L3 (UCH-L3) expression in GCSCs and GCICs was similar to that of β-catenin expression depending on glucose deprivation. We also observed that inhibition of UCH-L3 activity reduced β-catenin protein levels. The interaction between UCH-L3 and β-catenin proteins was confirmed, and it reduced the ubiquitination of β-catenin. Our results suggest that UCH-L3 induces the stabilization of β-catenin, which is required to promote stemness during autophagy activation. Also, UCH-L3 expression was regulated by c-Fos, and the levels of c-Fos increased in response to autophagy activation. In summary, our findings suggest that the inhibition of UCH-L3 during nutrient deprivation could suppress stress resistance of GCSCs and increase the survival rates of gastric cancer patients.

• Clinics in Shoulder and Elbow
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• 제25권4호
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• pp.296-303
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• 2022

Background: A previous study reported that hyperlipidemia increases the incidence of tears in the rotator cuff tendon and affects healing after repair. The aim of our study was to compare the gene and protein expression of torn rotator cuff tendons in patients both with and without hypercholesterolemia. Methods: Thirty patients who provided rotator cuff tendon samples were classified into either a non-hypercholesterolemia group (n=19, serum total cholesterol [TC] <200 mg/dL) and hypercholesterolemia group (n=11, serum TC ≥240 mg/dL) based on their concentrations of serum TC. The expression of various genes of interest, including COL1A1, IGF1, IL-6, MMP2, MMP3, MMP9, MMP13, TNMD, and TP53, was analyzed by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). In addition, Western blot analysis was performed on the proteins encoded by interleukin (IL)-6 and TP53 that showed significantly different expression levels in real-time qRT-PCR. Results: Except for IGF1, the gene expression levels of IL-6, MMP2, MMP9, and TP53 were significantly higher in the hypercholesterolemic group than in the non-hypercholesterolemia group. Western blot analysis confirmed significantly higher protein levels of IL-6 and TP53 in the hypercholesterolemic group (p<0.05). Conclusions: We observed an increase in inflammatory cytokine and matrix metalloproteinase (MMP) levels in hypercholesterolemic patients with rotator cuff tears. Increased levels of IL-6 and TP53 were observed at both the mRNA and protein levels. We suggest that the overexpression of IL-6 and TP53 may be a specific feature in rotator cuff disease patients with hypercholesterolemia.

• 한국치위생학회지
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• 제23권4호
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• pp.209-218
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• 2023

연구목적: 실험실내에서 성분이 다른 구강세정제가 오래 형성된 다중 구강 바이오필름에 미치는 항균효과를 확인하였다. 연구방법: 구강세정제의 항균효과를 ATP, CFU로 확인하였고, 다중 구강 바이오필름을 형성시킨 sHA 디스크를 구강세정제로 처리한 후 SEM을 이용하여 형태학적 모양을 관찰하였다. 총RNA는 P. intermedia의 바이오필름에서 추출하였고 RNA sequencing 분석을 하였다. 연구결과: 대조군과 구강세정제 4개를 비교한 CFU 측정 결과에서 오래 형성된 다중 구강 바이오필름은 39.0%로부터 95.7%까지 감소한 것으로 나타났다(p<0.05). 에센셜 오일 처리 후 구강생물막 세포들은 크기가 변하고 일부 파열되고 작은 세포 파편 조각도 나타났다. 에센셜 오일로 처리 후 P. intermedia 유전자 발현은 RNA 전사와 단백질 번역 관련 유전자에서 유의한 변화를 보였다. 결론: 성분이 다른 구강세정제는 항균효과를 가지며 표면 구조 및 유전자 발현에 영향을 미친다.

• Mycobiology
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• 제51권3호
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• pp.115-121
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• 2023

The fungal strain KNUF-22-18B, belonging to Cucurbitariaceae, was discovered from a stink bug (Hygia lativentris) during the investigation of insect microbiota in Chungnam Province, South Korea. The colonies of the strain KNUF-22-18B were wooly floccose, white to brown in the center on oatmeal agar (OA), and the colonies were buff, margin even, and colorless, reverse white to yellowish toward the center on malt extract agar (MEA). The strain KNUF-22-18B produced pycnidia after 60 days of culturing on potato dextrose agar, but pycnidia were not observed on OA. On the contrary, N. keratinophila CBS 121759^T abundantly formed superficial pycnidia on OA and MEA after a few days. The strain KNUF-22-18B produced chlamydospores subglobose to globose, mainly in the chain, with a small diameter of 4.4-8.8 ㎛. At the same time, N. keratinophila CBS 121759^T displayed a globose terminal with a diameter of 8-10 ㎛. A multilocus phylogeny using the internal transcribed spacer regions, 28S rDNA large subunit, b-tubulin, and RNA polymerase II large subunit genes further validated the uniqueness of the strain. The detailed description and illustration of the proposed species as Neocucurbitaria chlamydospora sp. nov. from Korea was strongly supported by molecular phylogeny.

• Mycobiology
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• 제51권2호
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• pp.87-93
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• 2023

The fungal strain belonging to the genus Monochaetia of the family Sporocadaceae was isolated from hairy long-horned toad beetle (Moechotypa diphysis) during the screening of microfungi associated with insects from Gangwon Province, Korea. The strain KNUF-6L2F produced white, light brown to dirty black surface, and olivaceous green colonies with the higher growth, while the closest strain M. ilicis KUMCC 15-0520^T were light brown to brown, and M. schimae SAUCC 212201^T light brown to brown toward center. The strain KNUF-6L2F produced shorter (5.7-14.0 ㎛) apical appendages than M. ilicis (6.0-24.0 ㎛), but similar to M. schimae (7.0-12.5 ㎛). Three median cells of KNUF-6L2F were light brown to olivaceous green, whereas brown and olivaceous cells were observed from M. ilicis and M. schimae, respectively. And the strain KNUF-6L2F produced larger conidiogenous cells than M. ilicis and M. schimae. Additionally, phylogenetic analyses based on molecular datasets of internal transcribed spacer (ITS) regions, translation elongation factor 1-alpha (TEF1α), and β-tubulin (TUB2) genes corroborated the strain's originality. Thus, the strain is different from other known Monochaetia species, according to molecular phylogeny and morophology, hence we suggested the new species Monochaetia mediana sp. nov. and provided a descriptive illustration.

• BMB Reports
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• 제56권8호
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• pp.463-468
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• 2023

Screening for genetic defects in the cells should be examined for clinical application. The Pearson syndrome (PS) patient harbored nuclear mutations in the POLG and SSBP1 genes, which could induce systemic large-scale mitochondrial genome (mtDNA) deletion. We investigated iPSCs with mtDNA deletions in PS patient and whether deletion levels could be maintained during differentiation. The iPSC clones derived from skin fibroblasts (9% deletion) and blood mononuclear cells (24% deletion) were measured for mtDNA deletion levels. Of the 13 skin-derived iPSC clones, only 3 were found to be free of mtDNA deletions, whereas all blood-derived iPSC clones were found to be free of deletions. The iPSC clones with (27%) and without mtDNA deletion (0%) were selected and performed in vitro and in vivo differentiation, such as embryonic body (EB) and teratoma formation. After differentiation, the level of deletion was retained or increased in EBs (24%) or teratoma (45%) from deletion iPSC clone, while, the absence of deletions showed in all EBs and teratomas from deletion-free iPSC clones. These results demonstrated that non-deletion in iPSCs was maintained during in vitro and in vivo differentiation, even in the presence of nuclear mutations, suggesting that deletion-free iPSC clones could be candidates for autologous cell therapy in patients.

• Journal of Korean Neurosurgical Society
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• 제66권5호
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• pp.525-535
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• 2023

Objective : We performed an expanded multi-ethnic meta-analysis to identify associations between inflammation-related loci with intracranial aneurysm (IA) susceptibility. This meta-analysis possesses increased statistical power as it is based on the most data ever evaluated. Methods : We searched and reviewed relevant literature through electronic search engines up to August 2022. Overall estimates were calculated under the fixed- or random-effect models using pooled odds ratio (OR) and 95% confidence intervals (CIs). Subgroup analyses were performed according to ethnicity. Results : Our meta-analysis enrolled 15 studies and involved 3070 patients and 5528 controls including European, Asian, Hispanic, and mixed ethnic populations. Of 17 inflammation-related variants, the rs1800796 locus (interleukin [IL]-6) showed the most significant genome-wide association with IA in East-Asian populations, including 1276 IA patients and 1322 controls (OR, 0.65; 95% CI, 0.56-0.75; p=3.24#x00D7;10^-9) under a fixed-effect model. However, this association was not observed in the European population (OR, 1.09; 95% CI, 0.80-1.47; p=0.5929). Three other variants, rs16944 (IL-1β), rs2195940 (IL-12B), and rs1800629 (tumor necrosis factor-α) showed a statistically nominal association with IA in both the overall, as well as East-Asian populations (0.01<p<0.05). Conclusion : Our updated meta-analysis with increased statistical power highlights that rs1800796 which maps on the IL-6 gene is associated with IA, and in particular confers a protective effect against occurrence of IA in the East-Asian population.

• Applied Microscopy
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• 제51권
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• pp.4.1-4.12
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• 2021

Fluorescence in situ hybridization (FISH) is a technique to visualize specific DNA/RNA sequences within the cell nuclei and provide the presence, location and structural integrity of genes on chromosomes. A confocal Whole Slide Imaging (WSI) scanner technology has superior depth resolution compared to wide-field fluorescence imaging. Confocal WSI has the ability to perform serial optical sections with specimen imaging, which is critical for 3D tissue reconstruction for volumetric spatial analysis. The standard clinical manual scoring for FISH is labor-intensive, time-consuming and subjective. Application of multi-gene FISH analysis alongside 3D imaging, significantly increase the level of complexity required for an accurate 3D analysis. Therefore, the purpose of this study is to establish automated 3D FISH scoring for z-stack images from confocal WSI scanner. The algorithm and the application we developed, SHIMARIS PAFQ, successfully employs 3D calculations for clear individual cell nuclei segmentation, gene signals detection and distribution of break-apart probes signal patterns, including standard break-apart, and variant patterns due to truncation, and deletion, etc. The analysis was accurate and precise when compared with ground truth clinical manual counting and scoring reported in ten lymphoma and solid tumors cases. The algorithm and the application we developed, SHIMARIS PAFQ, is objective and more efficient than the conventional procedure. It enables the automated counting of more nuclei, precisely detecting additional abnormal signal variations in nuclei patterns and analyzes gigabyte multi-layer stacking imaging data of tissue samples from patients. Currently, we are developing a deep learning algorithm for automated tumor area detection to be integrated with SHIMARIS PAFQ.

• The Plant Pathology Journal
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• 제39권5호
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• pp.430-448
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• 2023

Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.