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Comparison of the spatial-temporal distributions of the heterotrophic dinoflagellates Gyrodinium dominans, G. jinhaense, and G. moestrupii in Korean coastal waters

  • Lee, Sung Yeon;Jeong, Hae Jin;Kang, Hee Chang;Ok, Jin Hee;You, Ji Hyun;Park, Sang Ah;Eom, Se Hee
    • ALGAE
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    • v.36 no.1
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    • pp.37-50
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    • 2021
  • Heterotrophic dinoflagellates Gyrodinium spp. are one of the major grazers of phytoplankton in many coastal waters. Gyrodinium dominans, G. jinhaense, and G. moestrupii have similar morphologies but different edible prey species. To explore the variations in the ecological niches of these three species, we investigated their spatial-temporal distributions in Korean waters. Because of the high similarity in morphology among these three Gyrodinium species, we used real-time polymerase chain reactions to quantify their abundance in water samples that were seasonally collected from 28 stations along the Korean Peninsula from April 2015 to October 2018. Cells of G. dominans were found at all sampling stations, G. jinhaense at 26 stations, and G. moestrupii at 22 stations, indicating that all three species were widely distributed in Korea. Furthermore, all three species displayed strong seasonal distributions. The largest numbers of the stations where G. dominans and G. jinhaense cells were present were found during the summer (26 and 23 stations, respectively), but that for G. moestrupii was found in the autumn (15 stations). The abundance of G. dominans was positively correlated with that of G. jinhaense, but not with that of G. moestrupii. The highest abundances of G. dominans (202.5 cells mL-1) and G. jinhaense (20.2 cells mL-1) were much greater than that of G. moestrupii (1.2 cells mL-1). The highest abundances of G. dominans and G. jinhaense were found in July, whereas that of G. moestrupii was found in March. The abundances of G. dominans and G. jinhaense, but not G. moestrupii, were positively correlated with water temperature. Therefore, the spatial-temporal distributions of G. dominans and G. jinhaense were closer than those of G. moestrupii and G. dominans or G. jinhaense. This differs from results based on the relative differences in ribosomal DNA sequences and the types of edible prey reported in the literature. Thus, the variations in spatial-temporal distributions and prey species of these three Gyrodinium species suggest that they may have different ecological niches in Korean coastal waters.

Relationship between DNA mismatch repair and CRISPR/Cas9-mediated knock-in in the bovine β-casein gene locus

  • Kim, Seung-Yeon;Kim, Ga-Yeon;You, Hyeong-Ju;Kang, Man-Jong
    • Animal Bioscience
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    • v.35 no.1
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    • pp.126-137
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    • 2022
  • Objective: Efficient gene editing technology is critical for successful knock-in in domestic animals. RAD51 recombinase (RAD51) gene plays an important role in strand invasion during homologous recombination (HR) in mammals, and is regulated by checkpoint kinase 1 (CHK1) and CHK2 genes, which are upstream elements of RAD51 recombinase (RAD51). In addition, mismatch repair (MMR) system is inextricably linked to HR-related pathways and regulates HR via heteroduplex rejection. Thus, the aim of this study was to investigate whether clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9)-mediated knock-in efficiency of human lactoferrin (hLF) knock-in vector in the bovine β-casein gene locus can be increased by suppressing DNA MMR-related genes (MSH2, MSH3, MSH6, MLH1, and PMS2) and overexpressing DNA double-strand break (DSB) repair-related genes (RAD51, CHK1, CHK2). Methods: Bovine mammary epithelial (MAC-T) cells were transfected with a knock-in vector, RAD51, CHK1, or CHK2 overexpression vector and CRISPR/sgRNA expression vector to target the bovine β-casein gene locus, followed by treatment of the cells with CdCl2 for 24 hours. After 3 days of CdCl2 treatment, the knock-in efficiency was confirmed by polymerase chain reaction (PCR). The mRNA expression levels of DNA MMR-related and DNA DSB repair-related genes were assessed by quantitative real-time PCR (RT-qPCR). Results: Treatment with CdCl2 decreased the mRNA expression of RAD51 and MMRrelated genes but did not increase the knock-in efficiency in MAC-T cells. Also, the overexpression of DNA DSB repair-related genes in MAC-T cells did not significantly affect the mRNA expression of MMR-related genes and failed to increase the knock-in efficiency. Conclusion: Treatment with CdCl2 inhibited the mRNA levels of RAD51 and DNA MMR-related genes in MAC-T cells. However, the function of MMR pathway in relation to HR may differ in various cell types or species.

Gaseous signal molecule SO2 regulates autophagy through PI3K/AKT pathway inhibits cardiomyocyte apoptosis and improves myocardial fibrosis in rats with type II diabetes

  • Zhao, Junxiong;Wu, Qian;Yang, Ting;Nie, Liangui;Liu, Shengquan;Zhou, Jia;Chen, Jian;Jiang, Zhentao;Xiao, Ting;Yang, Jun;Chu, Chun
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.6
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    • pp.541-556
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    • 2022
  • Myocardial fibrosis is a key link in the occurrence and development of diabetic cardiomyopathy. Its etiology is complex, and the effect of drugs is not good. Cardiomyocyte apoptosis is an important cause of myocardial fibrosis. The purpose of this study was to investigate the effect of gaseous signal molecule sulfur dioxide (SO2) on diabetic myocardial fibrosis and its internal regulatory mechanism. Masson and TUNEL staining, Western-blot, transmission electron microscopy, RT-qPCR, immunofluorescence staining, and flow cytometry were used in the study, and the interstitial collagen deposition, autophagy, apoptosis, and changes in phosphatidylinositol 3-kinase (PI3K)/AKT pathways were evaluated from in vivo and in vitro experiments. The results showed that diabetic myocardial fibrosis was accompanied by cardiomyocyte apoptosis and down-regulation of endogenous SO2-producing enzyme aspartate aminotransferase (AAT)1/2. However, exogenous SO2 donors could up-regulate AAT1/2, reduce apoptosis of cardiomyocytes induced by diabetic rats or high glucose, inhibit phosphorylation of PI3K/AKT protein, up-regulate autophagy, and reduce interstitial collagen deposition. In conclusion, the results of this study suggest that the gaseous signal molecule SO2 can inhibit the PI3K/AKT pathway to promote cytoprotective autophagy and inhibit cardiomyocyte apoptosis to improve myocardial fibrosis in diabetic rats. The results of this study are expected to provide new targets and intervention strategies for the prevention and treatment of diabetic cardiomyopathy.

Impact of ZrO2 nanoparticles addition on flexural properties of denture base resin with different thickness

  • Albasarah, Sara;Al Abdulghani, Hanan;Alaseef, Nawarah;al-Qarni, Faisal D.;Akhtar, Sultan;Khan, Soban Q.;Ateeq, Ijlal Shahrukh;Gad, Mohammed M.
    • The Journal of Advanced Prosthodontics
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    • v.13 no.4
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    • pp.226-236
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    • 2021
  • PURPOSE. This study aimed to evaluate the effect of incorporating zirconium oxide nanoparticles (nano-ZrO2) in polymethylmethacrylate (PMMA) denture base resin on flexural properties at different material thicknesses. MATERIALS AND METHODS. Heat polymerized acrylic resin specimens (N = 120) were fabricated and divided into 4 groups according to denture base thickness (2.5 mm, 2.0 mm, 1.5 mm, 1.0 mm). Each group was subdivided into 3 subgroups (n = 10) according to nano-ZrO2 concentration (0%, 2.5%, and 5%). Flexural strength and elastic modulus were evaluated using a three-point bending test. One-way ANOVA, Tukey's post hoc, and two-way ANOVA were used for data analysis (α = .05). Scanning electron microscopy (SEM) was used for fracture surface analysis and nanoparticles distributions. RESULTS. Groups with 0% nano-ZrO2 showed no significant difference in the flexural strength as thickness decreased (P = .153). The addition of nano-zirconia significantly increased the flexural strength (P < .001). The highest value was with 5% nano-ZrO2 and 2 mm-thickness (125.4 ± 18.3 MPa), followed by 5% nano-ZrO2 and 1.5 mm-thickness (110.3 ± 8.5 MPa). Moreover, the effect of various concentration levels on elastic modulus was statistically significant for 2 mm thickness (P = .001), but the combined effect of thickness and concentration on elastic modulus was insignificant (P = .10). CONCLUSION. Reinforcement of denture base material with nano-ZrO2 significantly increased flexural strength and modulus of elasticity. Reducing material thickness did not decrease flexural strength when nano-ZrO2 was incorporated. In clinical practice, when low thickness of denture base material is indicated, PMMA/nano-ZrO2 could be used with minimum acceptable thickness of 1.5 mm.

Comparisons of Foliar Nutrient Concentrations and Soil Nutrient Stocks Between Restoration Areas and Adjacent Red Pine Stands in Opencast Kaolinite Mines in Sancheong-gun (산청군 고령토 광산 복원지와 인접 소나무 임분의 토양양분 저장량 및 잎 양분 농도 비교)

  • Kim, Kyung Tae;Kim, Choonsig
    • Journal of Korean Society of Forest Science
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    • v.111 no.2
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    • pp.234-241
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    • 2022
  • We performed this study to determine the foliar nutrient concentration and the nutrient stocks of restoration areas and adjacent Pinus densiflora S. et. Z. (red pine) stands in opencast kaolinite mines in Sancheong-gun, Gyeongsangnam-do, southern Korea. We chose six sites to determine foliage nutrient concentrations and the nutrient stocks of soils (0-10 cm depth). The dominant vegetation planted in restoration areas comprised Quercus acutissima Carruth., P. koraiensis S. et. Z., Festuca arundinacea Schreb., and Lespedeza cuneata G. Don. Invading vegetation in the restoration areas comprised Alnus incana (L.) Medik., Robinia pseudoacacia L., and Lespedeza spp., among others. The carbon and nitrogen stocks at 10 cm soil depth were significantly higher in the red pine stands than those in the restoration areas, whereas those of phosphorus, potassium, and magnesium were not significantly different between the two areas. However, calcium stocks were significantly higher in the restoration areas than in the red pine stands. Nitrogen concentration in foliage was higher in L. cuneata (20.28 mg N g-1) than that in F. arundinacea (5.67 mg N g-1), whereas potassium concentration was twice as high in F. arundinacea (18.8 mg K g-1) as that in L. cuneata (9.07 mg K g-1). Foliar nitrogen concentrations in invasive vegetation such as A. incana, R. pseudoacacia, and Lespedeza spp. were twice or four times higher than those of Q. acutissima and P. koraiensis. Our results indicate the development of suitable vegetation and soil amendment treatments to improve poor soil environmental conditions in restoration areas are necessary following opencast kaolinite mining.

Monitoring the Restoration of Evergreen Broad-Leaved Forests in the Warm-Temperate Region(III) (난온대 기후대의 상록활엽수림 복원 모니터링(III))

  • Kang, Hyun-Mi;Kang, Ji-Woo;Kim, Ji-Hae;Sung, Chan-Yong;Park, Seok-Gon
    • Korean Journal of Environment and Ecology
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    • v.36 no.1
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    • pp.87-101
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    • 2022
  • This study analyzed changes in the vegetation structure of 16 permanent plots (experimental and control) installed in Wando Arboretum in 1996 to monitor the long-term change process of evergreen broad-leaved forests in warm temperate. Especially, this study discusses the effects of trial treatment (selection cutting and plant introduction), introduced as a restoration technique in 1996, on vegetation development. In communities dominated by Quercus acuta in the canopy (permanent plots 1 through 3), this species's mean important percentage (MIP) decreased, and the evergreen broad-leaved species was introduced from outside increased, likely to change the vegetation structure in the future. The expansion of the evergreen broad-leaved species group was also confirmed in Q. acuta-deciduous broad-leaved forests (permanent plots 5 and 7) and Pinus densifloraforests (permanent plots 9 and 10). In the experimental plots where thinning was carried out, the zoochory (the dispersal of seeds by birds), Cinnamomum yabunikkei, Neolitsea sericea, Machilus thunbergii, etc., and the expansion of the influence of evergreen broad-leaved species were remarkable, so it is considered to have effectively promoted the vegetation development in warm temperate forests. Although evergreen broad-leaved species were planted in the experimental plot to change vegetation structure, it seems the effect on the change was weak due to the small amount of planting. Compared to other vegetation types, the change in the vegetative structure of the pine forest to an evergreen broad-leaved forest was clear due to the decline of P. densiflora and P. thunbergii.

Reexamination of Coach-Athlete Relationship Maintenance Scale in Pro Baseball (프로야구 코치-선수관계 유지 척도 재검증)

  • Huh, Jin-Young;Choi, Hun-Hyuk
    • 한국체육학회지인문사회과학편
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    • v.55 no.1
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    • pp.221-233
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    • 2016
  • The purpose of this study was to prove a development and initial validation of the korean version of coach-athlete relationship maintenance scale that originated from the work of Rhind & Jowett(2012) in pro baseball. The items were then administered to 132 Participants(29 coaches and 103 athletes) completed the questionnaires of the coach-athlete relationship maintenance in First preliminary investigation. Maximum likelihood estimate was used to identify the latent underlying structure. In order to verify the validity of Korean version of coach-athlete relationship maintenance was administered to an independent sample of 273 coaches and athletes. Pro baseball coach-athlete relationship maintenance is consisted of six factors(25 items) with conflict management, motivational, preventative, openness/assurance, support, and social network. SPSS18.0 and AMOS16.0 were used to analyze the exploratory factor analysis, confirmatory factory analysis and internal consistency, test-retest with bootstrapping using of the data in this study. The results of the pro baseball coach-athlete relationship maintenance scale had six factors with 25 items, and each six factor was positively correlated. Overall, this study verified pro baseball coach-athlete relationship maintenance questionnaire. Thus, suggest that path of comparing the differences between the first division and farm team by using the test of the structural model invariance across the groups.

Persistence Study of Thiamethoxam and Its Metabolite in Kiwifruit for Establishment of Import Tolerance

  • Il Kyu Cho;Gyeong Hwan Lee;Woo Young Cho;Yun-Su Jeong;Danbi Kim;Kil Yong Kim;Gi-Woo Hyoung;Chul Hong Kim
    • Korean Journal of Environmental Agriculture
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    • v.41 no.4
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    • pp.355-364
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    • 2022
  • BACKGROUND: Pre-harvest interval and decline pattern of thiamethoxam were determined in kiwifruit using liquid chromatography-tandem mass spectrometry (LCMS/MS). The study was carried out to propose import tolerance using OECD maximum residue limit (MRL) calculator for the export promotion of kiwifruit to Taiwan. METHODS AND RESULTS: The thiamethoxam residue in kiwifruit was determined by using the LC-TriQ-MS/MS with the analytical process to set up the import tolerance under greenhouse conditions for Taiwan. Excellent linearity was observed for all of the analytes with a determination coefficient (R2)≥0.99. The limit of quantification was determined to be 0.01 mg/kg for both thiamethoxam and clothianidin in kiwifruit. Linearity was determined from the co-efficient of determinants (R2) obtained from the seven-point calibration curve. The standard calibration curve showed as follows; 1) Site 1 (Gimje): y = 944,406X + 1,583 (R2=0.9995), 2) Site 2 (Goheung): y = 1,356,205X + 934 (R2=0.9983), and 3) Site 3 (Jangheung): y = 1,239,937X - 3,090 (R2=0.9908). The residue of thiamethoxam in the kiwifruit for three decline trials showed the range of 0.35 to 0.56 mg/kg in site 1 (Gimje), 0.24 to 0.55 mg/kg in site 2 (Goheung), and 0.28 to 0.42 mg/kg in site 3 (Jangheung), respectively. However, clothianidin was not detected in all of the treatments. The maximum residual amounts (decline) in the samples, sprayed according to the safe-use standard for thiamethoxam 10% WG in kiwifruit (30 days before harvest, 3 sprays every 7 days) were 0.56 mg/kg in site 1, 0.55 mg/kg in site 2, and 0.42 mg/kg in site 3, respectively. CONCLUSION(S): The import tolerance (IT) of thiamethoxam for kiwifruit may be proposed to be 0.9 mg/kg by using the OECD MRL calculator.

Scavenging Capacities of DPPH and ABTS Free Radicals and Anti-inflammatory Activities of Ethanol Extracts and their Fractions from Sophora tonkinensis

  • Eun Sun Moon;Ji Yoon Lee;Seongdae Kim;Chang Won Choi
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.46-46
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    • 2021
  • The first purpose of this study was to evaluate the scavenging capacity (SC) of DPPH and ABTS free radicals for ethanol extract (STR-E) and its active fractions from Sophora tonkinensis root (STR). Four different fractions from STR-E were prepared by using different types of solvents such as chloroform (STR-E-C), ethyl acetate (STR-E-EA), n-butanol (STR-E-B), and water (STR-E-W). STR-E-C showed the highest value of total phenolic content, while STR-E showed the highest value of total flavonoid and terpenoid content. In STR-E and its four fractions, STR-E-EA showed the strongest SC with the lowest SC50 values of the DPPH radicals and ABTS radicals. The second purpose of this study was to evaluate anti-inflammatory activity in the lipopolysaccharide (LPS)-induced RAW 264.7 macrophages treated with STR-E, STR-E-C, and STR-E-EA, respectively. No cytotoxic effect to RAW 264.7 cells was observed at 20 ~ 25 ㎍/ml of STR-E, 10 ㎍/ml of STR-E-C, and 5 ㎍/ml of the STR-E-EA, presenting cell viability values close to that of the untreated control (100%). STR-E, STR-E-C, and STR-E-EA significantly suppressed the LPS-induced nitric oxide (NO) in a dose-dependent manner. Results of reverse-transcription (RT)-qPCR analysis showed that the peak mRNA levels of IL-1β, TNF-α, iNOS, IL-6, and IL-10 were observed in the LPS-stimulated macrophages at 4 h, 2 h, 12 h, 12 h, and 12 h, respectively. The peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 were significantly reduced in the LPS-stimulated macrophages co-treated with 20 ㎍/ml and 25 ㎍/ml of STR-E, respectively. In the case of IL-10, its peak mRNA level slightly increased without statistical significance. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 10 ㎍/ml and 20 ㎍/ml of STR-E-C, respectively. In contrast, the peak mRNA level of IL-10 significantly increased at 8 h. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 5 ㎍/ml and 10 ㎍/ml of STR-E-EA, respectively. In contrast, the peak mRNA level of IL-10 increased at 4 h. Taken together, our data indicated that STR-E, STR-E-C, and STR-E-EA activate macrophages to secrete both pro-inflammatory and anti-inflammatory cytokines.

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Imaging-Based Versus Pathologic Survival Stratifications of Diffuse Glioma According to the 2021 WHO Classification System

  • So Jeong Lee;Ji Eun Park;Seo Young Park;Young-Hoon Kim;Chang Ki Hong;Jeong Hoon Kim;Ho Sung Kim
    • Korean Journal of Radiology
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    • v.24 no.8
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    • pp.772-783
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    • 2023
  • Objective: Imaging-based survival stratification of patients with gliomas is important for their management, and the 2021 WHO classification system must be clinically tested. The aim of this study was to compare integrative imaging- and pathology-based methods for survival stratification of patients with diffuse glioma. Materials and Methods: This study included diffuse glioma cases from The Cancer Genome Atlas (training set: 141 patients) and Asan Medical Center (validation set: 131 patients). Two neuroradiologists analyzed presurgical CT and MRI to assign gliomas to five imaging-based risk subgroups (1 to 5) according to well-known imaging phenotypes (e.g., T2/FLAIR mismatch) and recategorized them into three imaging-based risk groups, according to the 2021 WHO classification: group 1 (corresponding to risk subgroup 1, indicating oligodendroglioma, isocitrate dehydrogenase [IDH]-mutant, and 1p19q-codeleted), group 2 (risk subgroups 2 and 3, indicating astrocytoma, IDH-mutant), and group 3 (risk subgroups 4 and 5, indicating glioblastoma, IDHwt). The progression-free survival (PFS) and overall survival (OS) were estimated for each imaging risk group, subgroup, and pathological diagnosis. Time-dependent area-under-the receiver operating characteristic analysis (AUC) was used to compare the performance between imaging-based and pathology-based survival model. Results: Both OS and PFS were stratified according to the five imaging-based risk subgroups (P < 0.001) and three imaging-based risk groups (P < 0.001). The three imaging-based groups showed high performance in predicting PFS at one-year (AUC, 0.787) and five-years (AUC, 0.823), which was similar to that of the pathology-based prediction of PFS (AUC of 0.785 and 0.837). Combined with clinical predictors, the performance of the imaging-based survival model for 1- and 3-year PFS (AUC 0.813 and 0.921) was similar to that of the pathology-based survival model (AUC 0.839 and 0.889). Conclusion: Imaging-based survival stratification according to the 2021 WHO classification demonstrated a performance similar to that of pathology-based survival stratification, especially in predicting PFS.