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Effects of Epidermal Growth Factor (EGF) and Insulin-like Growth Factor-1 (IGF-1) on Maturation of Bovine Follicular Oocytes In Vitro (Epidermal Growth Factor(EGF)와 Insulin-like Growth Factor-1(IGF-1)이 소 난포란의 체외성숙에 미치는 영향)

  • 윤종택;정영호;한기영;최선호
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.245-249
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    • 1998
  • The purpose of this study was to evaluate the effects of growth factors such as epidermal growth factor (EGF) and insulin-like growth factor-1 (IGF-1) on maturation of bovine follicular oocytes in vitro. Oocytes were recovered from the ovaries of slaughtered Hanwoos. The oocytes were matured in TCM 199 at 39$^{\circ}C$, 5% $CO_2$ in air. Growth factors were added to maturation medium as follows: control (no serum), EGF (10ng/m1, 50ng/ml or 100ng/m1), IGF-1 (100ng/m1) and EGF (50ng/ml) + IGF-1 (100ng/m1). The oocytes were placed onto a slide and stained with aceto-orcein dye. Nuclear maturation was evaluated and classified as germinal vesicle breakdown (GVBD), metaphase-I (MI) or metaphase-ll(Mll). Maturation rates were 37.9% (control), 45.8% (EGF, 10ng/m1), 55.8% (EGF, 50ng/ml), 44.4% (EGF, 100ng/m1), 46.7% (IGF-1, 100ng/m1) and 67.0% (IGF-1+EGF). The highest group developed to Mll stage was IGF-1+EGF treatment group (p<0.05). Therefore, nuclear maturation of bovine oocytes were affected by both of growth factors, and it seems to have a mutual activity between them.

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Characterization of a Putative F-box Motif in Ibd1p/Bfalp, a Spindle Checkpoint Regulator of Budding Yeast Saccharomyces cerevisiae

  • Lee, Kyum-Jung;Hyung-Seo;Kiwon Song
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.286-292
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    • 2001
  • During mitosis. the proper segregation of duplicated chromosomes is corrdinated by a spindle check-point. The bifurcated spindle checkpoint blocks cell cycle progression at metaphase by monitoring unattached kinetochores and inhibits mitotic exit in response to the misorientation of the mitotic spin- dle Ibd1p/Bfa1p is a spindle checkpoint regulator of budding yeast in the Bub2p checkpoint pathway for mitotic exit and its disruption abolishes mitotic arrest when proper organization of the mitotic spin-dls inhibited. Ibd1p/Bfa1p localizes to the spindle pole body, a microtublue-organizing center in yeast, and its overexpression arrests the cell cycle in 80% of cells with an enlarged budy at mitosis and in 20 % of cells with multiple buds. In this study, we found that the C-terminus of Ibd1p/Bfa1p phys-ically interacts with Skp1p, a key component of SCF (Skp1/cullin/F-box) complex for ubiquition-medi-ated proteolysis of cel cycle regulatores as well as an evolutionally conserved kinetochore protein for cell cycle progression. A putative F-box motif was found in the C-terminus of Ibd1p/Bfa1p and its function was investigated by making mutants of conserved residues in the motif. These Ibd1p/Bfa1p mutants of a putative F-box interacted with SKp1p in vitro by two-hybrid assays as wild type Ibd1p/Bfa1p. Also these Ibd1p/Bfa1p utants displayed the overexpression phenotypes of wild type Ibd1p, when over-expressed under inducible promoters . These results suggest that a putative F-box motif of Ibd1p/Bfa1p is not essential for the interaction with SKp1p and its function in mitotic exit and cytokinesis.

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The Effects of Plant Growth Substances on the Callus Induction and Multiple Shoot Formation of Korean Lawngrass( Zoysia japonica Steud.) (식물생장조절제 처리가 들잔디의 Callus유기 및 Multiple Shoots형성에 미치는 영향)

  • 심재성;김동찬;서병기
    • Asian Journal of Turfgrass Science
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    • v.8 no.3
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    • pp.137-147
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    • 1994
  • We have established a high-frequency plant regeneration system via organogenesis from mature seed of Korean lawngrass(Zoysia japonica Steud.). The effects of 2,4-dichiorophenoxy acetic acid (2,4-D), 6-furfuryl amino purine (kinetin), $\alpha$-naphthaiene acetic acid (NAA), N6-benzyl amino pu-rine (BAP), and casein hydrolysate (CR) on cailus induction and multiple shoot formation on ex-posure to light were evaluated. Callus produced on the Murashige and Skoog (MS) medium containing 2,4-D and kinetin had high organogenesis potency. A single addition of 1.0 mg $L-^1$ 2,4-D significantly induced callus. Also, 1.0 mg L-$^1$ 2,4-D, with the addition of 0.1 mg $L-^1$ kinetin highly enlianced callus induction. The trend of cailus induction was also found on mediurn containing 0.1 mg $L-^1$ BAP with 1.0 mg $L-^1$2,4-D, and 1 g $L-^1$ CR with the addition of 1.0 mg $L-^1$ 2,4-D. However, NAA was no effective on callus formation. The growth of root was significantly high in the presence of 0.1 mg $L-^1$ kinetin compared to other concentrations. Over 2 mg $L-^1$ kinetin highly lengthened roots. Fresh weight of plantlet was highest on medium containing 0.1 mg $L-^1$ 2,4-D. Also, on medium containing 0.1 mg $L-^1$ BAP, fresh weight of piantlet was highly enhanced. BAP was significantly effective on multiple shoot formation, particularly when 2.0 mg $L-^1$ was added with 0.1 mg $L-^1$ 2,4-D. Callus induction and multiple shoot formation were achieved on MS basal medium containing 1.0 g $L-^1$ CH.

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Effects of Nutrition Related Factors on Mortality Risk among Community-residing Older Adults in Korea (한국 지역사회 거주 노인의 영양관련 요인이 사망위험에 미치는 영향)

  • Lee, Si-Eun;Lee, Eliza
    • Journal of Digital Convergence
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    • v.16 no.10
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    • pp.343-350
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    • 2018
  • The purpose of this study was to identify the effect of nutrition related factors on mortality risk among community-residing older adults in Korea. We analyzed data from 8,532 older adults who took part in the 2008 living profiles of older people survey and 2011 death mortality data. According to Cox regression analysis, male (Hazard Ratio [HR], 2.03; 95% Confidence Interval [CI], 1.74~2.37), diabetes mellitus (HR, 1.35; 95% CI, 1.12~1.64), underweighted (HR, 2.68; 95% CI, 2.16~3.31), dysphagia (HR, 1.23; 95% CI, 1.02~1.47), chewing ability (HR, 1.33; 95% CI, 1.11~1.59), and undernutrition (HR, 1.47; 95% CI, 1.23~1.75) significantly affected mortality risk. Systematic nutrition support programs should be developed to decrease the mortality among Korean older adults.

Forecasting the Container Throughput of the Busan Port using a Seasonal Multiplicative ARIMA Model (승법계절 ARIMA 모형에 의한 부산항 컨테이너 물동량 추정과 예측)

  • Yi, Ghae-Deug
    • Journal of Korea Port Economic Association
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    • v.29 no.3
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    • pp.1-23
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    • 2013
  • This paper estimates and forecasts the container throughput of Busan port using the monthly data for years 1992-2011. To do this, this paper uses the several seasonal multiplicative ARIMA models. Among several ARIMA models, the seasonal multiplicative ARIMA model $(1,0,1){\times}(1,0,1)_{12}$ is selected as the best model by AIC, SC and Hannan-Quin information criteria. According to the forecasting values of the selected seasonal multiplicative ARIMA model $(1,0,1){\times}(1,0,1)_{12}$, the container throughput of Busan port for 2013-2020 will increase steadily annually, but there will be some volatile variations monthly due to the seasonality and other factors. Thus, to forecast the future container throughput of Busan port and to develop the Busan port efficiently, we need to use and analyze the seasonal multiplicative ARIMA model $(1,0,1){\times}(1,0,1)_{12}$.

Isolation, Purification, and Characterization of a Thermostable Xylanase from a Novel Strain, Paenibacillus campinasensis G1-1

  • Zheng, Hongchen;liu, Yihan;Liu, Xiaoguang;Wang, Jianling;Han, Ying;Lu, Fuping
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.930-938
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    • 2012
  • High levels of xylanase activity (143.98 IU/ml) produced by the newly isolated Paenibacillus campinasensis G1-1 were detected when it was cultivated in a synthetic medium. A thermostable xylanase, designated XynG1-1, from P. campinasensis G1-1 was purified to homogeneity by Octyl-Sepharose hydrophobic-interaction chromatography, Sephadex G75 gel-filter chromatography, and Q-Sepharose ion-exchange chromatography, consecutively. By multistep purification, the specific activity of XynG1-1 was up to 1,865.5 IU/mg with a 9.1-fold purification. The molecular mass of purified XynG1-1 was about 41.3 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Sequence analysis revealed that XynG1-1 containing 377 amino acids encoded by 1,134 bp genomic sequences of P. campinasensis G1-1 shared 96% homology with XylX from Paenibacillus campinasensis BL11 and 77%~78% homology with xylanases from Bacillus sp. YA-335 and Bacillus sp. 41M-1, respectively. The activity of XynG1-1 was stimulated by $Ca^{2+}$, $Ba^{2+}$, DTT, and ${\beta}$-mercaptoethanol, but was inhibited by $Ni^{2+}$, $Fe^{2+}$, $Fe^{3+}$, $Zn^{2+}$, SDS, and EDTA. The purified XynG1-1 displayed a greater affinity for birchwood xylan, with an optimal temperature of $60^{\circ}C$ and an optimal pH of 7.5. The fact that XynG1-1 is cellulose-free, thermostable (stability at high temperature of $70^{\circ}C{\sim}80^{\circ}C$), and active over a wide pH range (pH 5.0~9.0) suggests that the enzyme is potentially valuable for various industrial applications, especially for pulp bleaching pretreatment.

Evaluation of circulating PD-1 and PD-L1 as diagnostic biomarkers in dogs with tumors

  • Song, Doo-Won;Ro, Woong-Bin;Park, Hee-Myung
    • Journal of Veterinary Science
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    • v.22 no.5
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    • pp.75.1-75.10
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    • 2021
  • Background: Programmed cell death protein-1 (PD-1) and programmed cell death ligand-1 (PD-L1) have important roles in tumor evasion of the immune system. Objectives: This study aimed to assess the diagnostic utility of circulating PD-1 and PD-L1 levels in healthy dogs and dogs with tumors. Methods: Circulating PD-1 and PD-L1 levels in the serum of 71 dogs with tumors were compared with those of 52 healthy dogs by performing enzyme-linked immunosorbent assay (ELISA). Results: The ELISA results revealed higher circulating PD-1 and PD-L1 levels in dogs with tumors (2.9 [2.2-3.7] ng/mL; median [IQR] and 2.4 [1.4-4.4] ng/mL, respectively) than in healthy dogs (2.4 [1.9-3.0] ng/mL; p = 0.012 and 1.4 [0.9-2.1] ng/mL; p < 0.001, respectively). Especially, there was a significant difference in circulating PD-1 levels between healthy dogs and dogs with malignant epithelial tumors (2.4 [1.9-3.0] ng/mL and 3.1 [2.6-4.4] ng/mL, respectively; p < 0.01). In addition, there was a significant difference in circulating PD-L1 levels between healthy dogs and dogs with lymphomas (1.4 [0.9-2.1] ng/mL and 2.7 [1.6-5.8] ng/mL, respectively; p < 0.001). Conclusion: This study indicates that circulating PD-1 and PD-L1 have potential as tumor diagnostic biomarkers in dogs with tumors.

Inhibition of VRK1 suppresses proliferation and migration of vascular smooth muscle cells and intima hyperplasia after injury via mTORC1/β-catenin axis

  • Sun, Xiongshan;Zhao, Weiwei;Wang, Qiang;Zhao, Jiaqi;Yang, Dachun;Yang, Yongjian
    • BMB Reports
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    • v.55 no.5
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    • pp.244-249
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    • 2022
  • Characterized by abnormal proliferation and migration of vascular smooth muscle cells (VSMCs), neointima hyperplasia is a hallmark of vascular restenosis after percutaneous vascular interventions. Vaccinia-related kinase 1 (VRK1) is a stress adaption-associated ser/thr protein kinase that can induce the proliferation of various types of cells. However, the role of VRK1 in the proliferation and migration of VSMCs and neointima hyperplasia after vascular injury remains unknown. We observed increased expression of VRK1 in VSMCs subjected to platelet-derived growth factor (PDGF)-BB by western blotting. Silencing VRK1 by shVrk1 reduced the number of Ki-67-positive VSMCs and attenuated the migration of VSMCs. Mechanistically, we found that relative expression levels of β-catenin and effectors of mTOR complex 1 (mTORC1) such as phospho (p)-mammalian target of rapamycin (mTOR), p-S6, and p-4EBP1 were decreased after silencing VRK1. Restoration of β-catenin expression by SKL2001 and re-activation of mTORC1 by Tuberous sclerosis 1 siRNA (siTsc1) both abolished shVrk1-mediated inhibitory effect on VSMC proliferation and migration. siTsc1 also rescued the reduced expression of β-catenin caused by VRK1 inhibition. Furthermore, mTORC1 re-activation failed to recover the attenuated proliferation and migration of VSMC resulting from shVrk1 after silencing β-catenin. We also found that the vascular expression of VRK1 was increased after injury. VRK1 inactivation in vivo inhibited vascular injury-induced neointima hyperplasia in a β-catenin-dependent manner. These results demonstrate that inhibition of VRK1 can suppress the proliferation and migration of VSMC and neointima hyperplasia after vascular injury via mTORC1/β-catenin pathway.

LOXL1-AS1 Aggravates Myocardial Ischemia/Reperfusion Injury Through the miR-761/PTEN Axis

  • Wenhua He;Lili Duan;Li Zhang
    • Korean Circulation Journal
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    • v.53 no.6
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    • pp.387-403
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    • 2023
  • Background and Objectives: Myocardial ischemia and reperfusion injury (MIRI) has high morbidity and mortality worldwide. We aimed to explore the role of long noncoding RNA lysyl oxidase like 1 antisense RNA 1 (LOXL1-AS1) in cardiomyocyte pyroptosis. Methods: Hypoxia/reoxygenation (H/R) injury was constructed in human cardiomyocyte (HCM). The level of LOXL1-AS1, miR-761, phosphatase and tensin homolog (PTEN) and pyroptosis-related proteins was monitored by quantitative real-time polymerase chain reaction or western blot. Flow cytometry examined the pyroptosis level. Lactate dehydrogenase (LDH), creatine kinase-MB and cardiac troponin I levels were detected by test kits. Enzyme-linked immunosorbent assay measured the release of inflammatory cytokines. Dual-luciferase assay validated the binding relationship among LOXL1-AS1, miR-761, and PTEN. Finally, ischemia/reperfusion (I/R) animal model was constructed. Hematoxylin and eosin staining assessed morphological changes of myocardial tissue. NOD-like receptor pyrin domain-containing protein 3 (NLRP3) and casepase-1 expression was determined by immunohistochemistry. Results: After H/R treatment, LOXL1-AS1 and PTEN were highly expressed but miR-761 level was suppressed. LOXL1-AS1 inhibition or miR-761 overexpression increased cell viability, blocked the release of LDH and inflammatory cytokines (interleukin [IL]-1β, IL-18), inhibited pyroptosis level, and downregulated pyroptosis-related proteins (ASC, cleaved caspase-1, gasdermin D-N, NLRP3, IL-1β, and IL-18) levels in HCMs. LOXL1-AS1 sponged miR-761 to up-regulate PTEN. Knockdown of miR-761 reversed the effect of LOXL1-AS1 down regulation on H/R induced HCM pyroptosis. LOXL1-AS1 aggravated the MIRI by regulating miR-761/PTEN axis in vivo. Conclusions: LOXL1-AS1 targeted miR-761 to regulate PTEN expression, then enhance cardiomyocyte pyroptosis, providing a new alternative target for the treatment of MIRI.

GENETIC SUSCEPTIBILITIES OF CYTOCHROME P450 1A1, 2E1, AND N-ACETYLTRANSFERASE 2 TO THE RISKS FOR KOREAN HEAD AND NECK CANCER PATIENTS (한국인 두경부암종 환자에서 Cytochrome P450 1A1, 2E1 및 N-acetyltransferase 2 효소의 다형성 분석에 따른 유전적 감수성에 대한 연구)

  • Lee, Young-Soo;Kim, Te-Gyun;Woo, Soon-Seop;Shim, Kwang-Sub;Kong, Gu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.22 no.4
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    • pp.373-382
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    • 2000
  • Individual genetic susceptibilities to cancers may result from several factors including differences in xenobiotics metabolism to chemical carcinogens, DNA repair, altered oncogenes and suppressor genes, and environmental carcinogen exposures. Among them, genetic polymorphisms of metabolizing enzymes to chemical carcinogens have been recognized as a major important host factors in human cancers. They have two main types of enzymes: the phase I cytochrome P-450 mediating enzymes (CYPs) and phase II conjugating enzymes. The purpose of this study is to determine the frequencies of genotypes of phase I (CYP1A1 and CYP2E1) and phase II (NAT2) metabolizing enzymes in healthy control and head and neck cancer patients of Korean and to identify the relative high risk genotypes of these metabolizing enzymes to head and neck cancer in Korean. The author has analyzed 132 head and neck cancer patients and 113 healthy controls using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The results were as following; 1. The frequencies of genotypes of CYP1A1, CYP2E1 and NAT2 in healthy control were as following; CYP1A1 exon 7 polymorphism; Ile/Ile: Ile/Val: Val/Val = 59.3%: 36.3%: 4.4% CYP2E1 Pst I polymorphism, C1/C1: C1/C2: C2/C2 = 61.1%: 32.1%: 6.2% NAT2 polymorphism; F/F: F/S: S/S = 43.4%: 48.7%: 8.0% 2. In analysis of phase I enzyme, Val/Val genotype in CYP1A1 exon 7 polymorphism and C2/C2 genotype in CYP2E1 Pst I polymorphism were associated with relative high risks to head and neck cancers (Odds' ratio: 2.09 and 1.37, respectively). 3. Among the genotypes of NAT2 enzyme polymorphism, S/S genotype of NAT2 enzyme had 1.03 times of relative risk to head and neck cancers. 4. In combined genotyping of CYP1A1, CYP2E1, and NAT2 enzymes polymorphisms, the patients with Val/Val and C1/C1, C2/C2 and fast acetylator, and Val/Val and fast acetylator had higher relative risks than the patients with each baseline of combined genotypes (Odds' ratio: 2.82, 1.98 and 2.1, respectively). These results suggest the combined genotypes of Val/Val and C1/C1, C2/C2 and fast acetylator, and Val/Val and fast acetylator were more susceptible to head and neck cancers in Korean. And genotyping of metabolizing enzymes could be useful for predicting individual susceptibility to head and neck cancer.

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