Deep Seawater Increases Dendritic Branches of Cultured Rat Hippocampal Neurons

HyunSook Lee(이현숙),Kyung Soo Nam(남경수),Yun Hee Shon(손윤희),Il Soo Moon(문일수) 한국생명과학회 2008 생명과학회지 Vol.18 No.6

해양심층수(deep seawater, DSW)는 청정성과 무기물질의 풍부함 때문에 여러 분야에 응용하기 위하여 최근 많은 관심을 받고 있다. 본 연구에서는 동해 양양 부근의 해저 1,100 m에서 취수하여 역삼투압 시스템으로 탈염과 농축을 한 심층수가 배양한 흰쥐해마신경세포의 형태적 분화에 미치는 영향을 조사하였다. 10%(v/v) fetal bo-vine serum이 첨가된 MEM 배지에서 키운 세포와 비교할 때 25%(v/v) DSW이 포함될 경우 배양 17시간째에는 차이가 없었다. 그러나 DIV3, 7, 14, 및 17에 관찰하면 경도 0 및 200의 DSW가 포함된 배지에서 자란 신경세포는 가지돌기의 수가 현저히 줄었다. 반면에 경도 600의 DSW에서 자란 신경세포는 그 가지돌기의 수가 대조군과 비슷하였으며, 경도 1000의 경우는 대조군에 비하여 거의 2배 증가하였다. 이 결과는 적당한 경도의 DSW는 신경세포의 성장 및 건강을 돕는 것으로 해석된다. Deep seawater (DSW; deep ocean water) is pure, rich in inorganic materials which have attracted attention for various applications. In this study we investigated the effects of the DSW upwelled from the East Sea, offshore Yang Yang (Korea) on the morphological differentiation of cultured rat hippocampal neurons, which were grown in the minimal essential medium containing 10% (v/v) fetal bo-vine serum and 25% (v/v) DSW with various hardness. DSW had no effect on initial morphological differentiation (17 hr post-plating). When observed on DIV3, 7, 14, and 17, low hardness (0 and 200) DSW reduced dendritic branching. However, dendritic branches within 80 μm diameter from the center of soma nearly doubled in neurons grown in hardness 1,000 DSW-containing media. DSW with hardness 600 was more or less same as control groups. These results indicate that DSW with appropriate hardness ameliorates neuronal health.

From Epistemic Verbs to Concessives

Hyunsook Lee(이현숙) 담화·인지언어학회 2010 담화와 인지 Vol.17 No.3

In English, there are many markers to express concessive relationships. Among them, this paper aims at verbal origin concessives considering, seeing, granting, and supposing. They are all derived from verbs and acquire grammatical status as concessives. Users of language appear to opt for a different linguistic realization of the same concept (Hoffman 2005). Verbal origin concessives are not frequently used as much as although, though, in spite of, and a few others. However, because those items of content source have grammaticality and are used by speakers as one of the methods to denote the sense of concession, they should be shed light on in terms of grammaticalization. The present investigation, therefore, will look into the actual process in which English concessive markers derived from verbal origin are grammaticalized. First, items in question will be examined about grammaticality and the effect of analogy, and then diachronic semantic changes will be discussed through a perspective shift, subjectification and metonymy.

CT Examinations for COVID-19: A Systematic Review of Protocols, Radiation Dose, and Numbers Needed to Diagnose and Predict

Lee Jong Hyuk,Hong Hyunsook,Kim Hyungjin,Lee Chang Hyun,Goo Jin Mo,Yoon Soon Ho 대한영상의학회 2021 대한영상의학회지 Vol.82 No.6

Purpose Although chest CT has been discussed as a first-line test for coronavirus disease 2019 (COVID-19), little research has explored the implications of CT exposure in the population. To review chest CT protocols and radiation doses in COVID-19 publications and explore the number needed to diagnose (NND) and the number needed to predict (NNP) if CT is used as a first-line test. Materials and Methods We searched nine highly cited radiology journals to identify studies discussing the CT-based diagnosis of COVID-19 pneumonia. Study-level information on the CT protocol and radiation dose was collected, and the doses were compared with each national diagnostic reference level (DRL). The NND and NNP, which depends on the test positive rate (TPR), were calculated, given a CT sensitivity of 94% (95% confidence interval [CI]: 91%–96%) and specificity of 37% (95% CI: 26%–50%), and applied to the early outbreak in Wuhan, New York, and Italy. Results From 86 studies, the CT protocol and radiation dose were reported in 81 (94.2%) and 17 studies (19.8%), respectively. Low-dose chest CT was used more than twice as often as standarddose chest CT (39.5% vs.18.6%), while the remaining studies (44.2%) did not provide relevant information. The radiation doses were lower than the national DRLs in 15 of the 17 studies (88.2%) that reported doses. The NND was 3.2 scans (95% CI: 2.2–6.0). The NNPs at TPRs of 50%, 25%, 10%, and 5% were 2.2, 3.6, 8.0, 15.5 scans, respectively. In Wuhan, 35418 (TPR, 58%; 95% CI: 27710–56755) to 44840 (TPR, 38%; 95% CI: 35161–68164) individuals were estimated to have undergone CT examinations to diagnose 17365 patients. During the early surge in New York and Italy, daily NNDs changed up to 5.4 and 10.9 times, respectively, within 10 weeks. Conclusion Low-dose CT protocols were described in less than half of COVID-19 publications, and radiation doses were frequently lacking. The number of populations involved in a first-line diagnostic CT test could vary dynamically according to daily TPR; therefore, caution is required in future planning.

Identification of Potential Substrates of N-acetylglucosamine Kinase by a Proteomic Approach

HyunSook Lee(이현숙),Il Soo Moon(문일수) 한국생명과학회 2013 생명과학회지 Vol.23 No.4

단백질 번역 후 O-GlcNAc 수식은 단백질 조절의 새로운 기전으로 대두되고 있다. 전통적인 당수식과 달리 O-GlcNAc 수식은 단 한번의 O-GlcNAc 전달로 이루어지며, 핵 및 세포질단백질 모두에 수식될 수 있다. O-GlcNAc은 이 분자를 끝으로 하는 최종수식으로 생각되어 왔으나, 최근의 논문(J Proteome Res. 201110:2725-2733)은 AP180 단백질에 O-GlcNAc-P가 존재함을 보고하였다. 이 논문은 O-GlcNAc-P가 일반적인 단백질 수식인지에 대한 중요한 질문을 던진다. 이에 답하고자 저자들은 HEK293T 세포에 O-GlcNAc 인산화효소 NAGK를 DsRed2에 연결한 DsRed2-NAGKWT 혹은 효소활성이 없는 돌연변이 NAGK를 표현하는 DsRed2-NAGKD107A를 표현시키고, 단백질 추출물을 얻어 2D-PAGE로 분리한 후 인산화 정도를 측정하여, NAGKWT에 의하여 인산화가 증가되는 15개의 단백질 스폿을 선별하였다. 이 가운데 7개 스팟을 동정한 결과 2개의 스폿은 O-GlcNAc 수식 단백질인 HSP90β, 다른 2개의 스폿도 O-GlcNAc 수식 단백질인 ENO1로 동정되었으며, 나머지(dUTP nucleotidohydrolase mitochondrial isoform 2, glutathione S-transferase P, grp94)는 O-GlcNAc수식 여부를 아직 모르는 단백질이였다. NAGK에 의하여 O-GlcNAc 단백질의 인산화가 증가된다는 사실은 O-GlcNAc이 인산화되어 O-GlcNAc-P로 수식됨을 시사하며, 따라서 본 연구의 결과는 O-GlcNAc이 최종 수식이 아님을 지지한다. Post-translational O-GlcNAc modification (O-GlcNAcylation) of serine or threonine is a new protein modulation mechanism. In contrast to the classical glycosylation, O-GlcNAcylation occurs in a onestep transfer of O-GlcNAc on both nuclear and cytoplasmic proteins. In contrast to the general consensus that O-GlcNAc is a final modification, a recent paper (J Proteome Res. 2011 10:2725?2733) showed the presence of O-GlcNAc-P on a synaptic assembly protein AP180. This finding raises a fundamental question about its prevalence. To address this question, we used proteomics to identify those proteins that were phospho-signal enriched by GlcNAc kinase (NAGK). Comparison of pDsRed2-NAGKWT-transfected HEK293T cell extract with pDsRed2-NAGKD107A-transfected control culture revealed 15 phospho-signal increased spots. Excluding those spots that had no detectable amount of protein expression yielded 7 spots, which were selected for ID determination. Among these, two duplicate spots (two HSP90β and two ENO1 spots) were shown to be O-GlcNAcylated, two (dUTP nucleotidohydrolase mitochondrial isoform 2, glutathione S-transferase P) were not known to be involved in OGlcNAcylation, and one (heat shock protein gp96 precursor or grp94) was a glycoprotein. The increase in the phospho-levels of O-GlcNAc by NAGK strongly indicates that these proteins are phosphorylated on O-GlcNAc. Our present data support the idea that O-GlcNAc is not a terminal modification.

Advanced tube formation assay using human endothelial colony forming cells for in vitro evaluation of angiogenesis

Lee, Hyunsook,Kang, Kyu-Tae The Korean Society of Pharmacology 2018 The Korean Journal of Physiology & Pharmacology Vol.22 No.6

The tube formation assay is a widely used in vitro experiment model to evaluate angiogenic properties by measuring the formation of tubular structures from vascular endothelial cells (ECs). In vitro experimental results are crucial when considered the advisability of moving forward to in vivo studies. Thus, the additional attentions to the in vitro assay is necessary to improve the quality of the pre-clinical data, leading to better decision-making for successful drug discovery. In this study, we improved the tube formation assay system in three aspects. First, we used human endothelial colony forming cells (ECFCs), which are endothelial precursors that have a robust proliferative capacity and more defined angiogenic characteristics compared to mature ECs. Second, we utilized a real-time cell recorder to track the progression of tube formation for 48 hours. Third, to minimize analysis error due to the limited observation area, we used image-stitching software to increase the microscope field of view to a $2{\times}2$ stitched area from the $4{\times}$ object lens. Our advanced tube formation assay system successfully demonstrated the time-dependent dynamic progression of tube formation in the presence and absence of VEGF and FGF-2. Vatalanib, VEGF inhibitor, was tested by our assay system. Of note, $IC_{50}$ values of vatalanib was different at each observation time point. Collectively, these results indicate that our advanced tube formation assay system replicates the dynamic progression of tube formation in response to angiogenic modulators. Therefore, this new system provides a sensitive and versatile assay model for evaluating pro- or anti-angiogenic drugs.

Globalization and Korean History Education in English in Korea

Lee Injae,Lee Hyunsook The Academy of Korean Studies 2010 THE REVIEW OF KOREAN STUDIES Vol.13 No.2

Water Extract of Rhei Rhizoma Prevent Production of Reactive Oxygen Species and Loss of Mitochondrial Membrane Potential in a Hypoxia Model of Cultured Neurons

HyunSook Lee(이현숙),Il Soo Moon(문일수) 한국생명과학회 2008 생명과학회지 Vol.18 No.12

대황(Rhei Rhizoma; RR, 大黃)은 Rheum officinale Baill.와 Rheum palmatum L. (Polygonaceae)의 땅속부분(rhizome 및 root)으로 남아시아의 민속의학에서 간 및 신장의 손상을 치료하는데 널리 이용되고 있다. 본 연구에서는 배양한 흰쥐 해마신경세포의 저산소증모델을 이용하여 대황의 물추출물이 신경세포사를 억제하는 효능에 대하여 조사하였다. 배양 10일(DIV10)에 RR을 배양액에 첨가하고 DIV13일에 생존율을 조사한 결과 10 ㎍/㎖ 농도까지는 세포독성이 없었으며, 정상산소 환경에서 2.5 ㎍/㎖의 농도에서 세포생존율을 높이는 것으로 나타났다. 또한 배지에 대황을 첨가한 경우 DIV13일에 저산소증을 유도한 후 5일째에 세포생존율을 조사한 결과 대조군에 비하여 매우 유의하게 생존율을 증가시켰다. H₂DCF 염색 결과 대황은 활성산소(ROS)의 생성을 유의하게 감소시킴과, JC-1 염색 결과 사립체 막전위의 소실을 유의하게 억제함을 알 수 있었다. 이러한 결과들은 대황 물추출물이 활성산소를 효과적으로 제거하고, 세포의 에너지 생성을 보전함으로서 세포사를 억제할 수 있음을 보여주며, 향후 뇌신경 세포의 건강에 유용하게 이용될 수 있음을 시사한다. Rhei Rhizoma (RR; 大黃) consists of the underground parts (rhizome and root) of Rheum officinale Baill. and Rheum palmatum L. (Polygonaceae), and is widely used in Southeast Asian folk medicine to alleviate liver and kidney damages. In this study, we investigated into the efficacy and mechanism of RR water extract in supporting neuronal survival in a hypoxia model of cultured rat hippocampal neurons. RR exhibited no cytotoxicity up to 10 ㎍/㎖ and exhibited neurosupportive effects at 2.5 ㎍/㎖ in normoxia. When RR was added to the culture media on 10 days in vitro (DIV10) and given a hypoxic shock (2% O₂/5% CO₂, 3 hr, 37℃) on DIV13, RR exhibited neuroprotective effects on 5 days post-shock. H₂DCF stainings indicated that RR effectively prevents ROS production in both normoxia and hypoxia. JC-1 stainings showed that RR prevents dissipation of MMP in hypoxia. These results indicate that RR protects neurons by suppressing ROS production and MMP loss.

A Case Study on Teaching Korean History in English in Korea

Lee Hyunsook The Academy of Korean Studies 2010 THE REVIEW OF KOREAN STUDIES Vol.13 No.2

Preparation of a Monoclonal Antibody against Gintonin and Its Use in an Enzyme Immunoassay.

Lee, Byung-Hwan,Choi, Sun-Hye,Kim, Hyeon-Joong,Jung, Seok-Won,Kim, Hyunsook,Shin, Ho-Chul,Lee, Joon-Hee,Hwang, Sung-Hee,Kim, Hyoung-Chun,Nah, Seung-Yeol Pharmaceutical Society of Japan 2015 Biological & pharmaceutical bulletin Vol.38 No.10

<P>Gintonin is a novel ginseng-derived G protein-coupled lysophosphatidic acid (LPA) receptor ligand. Gintonin elicits an [Ca(2+)]i transient in animal cells via activation of LPA receptors. In vitro studies have shown that gintonin regulates various calcium-dependent ion channels and receptors. In in vivo studies, gintonin elicits anti-Alzheimer's disease activity through the activation of the non-amyloidogenic pathway and anti-metastatic effects through the inhibition of autotaxin. However, a method for gintonin quantitation in ginseng has not been developed. In the present study, we developed an enzyme immunoassay (EIA) to measure gintonin. A monoclonal antibody was raised in a mouse using gintonin as the immunogen, and an indirect competitive EIA was used to measure gintonin. The working range was 0.01-10???µg per assay. The anti-gintonin monoclonal antibody did not cross-react with the ginsenosides Ra, Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, and Rg3 or with LPAs such as LPA C16:0, LPA C18:0, LPA C18:1, and LPA C18:2. Using a standard curve, we measured the amount of gintonin in various ginseng extract fractions. Interestingly, we only detected a little amount of gintonin in conventional hot water extracts of Korean red ginseng. However, we can measure gintonin after ethanol extraction of Korean red ginseng marc. Thus, gintonin can be extracted from ginseng with ethanol but not water, and the remaining Korean red ginseng marc can be used to obtain gintonin. These results indicate that the EIA with the anti-gintonin monoclonal antibody can be used to quantify gintonin in various ginseng preparations, including commercial ginseng products.</P>

How Chromosome Mis-Segregation Leads to Cancer: Lessons from BubR1 Mouse Models

Lee, Hyunsook Korean Society for Molecular and Cellular Biology 2014 Molecules and cells Vol.37 No.10

Alteration in chromosome numbers and structures instigate and foster massive genetic instability. As Boveri has seen a hundred years ago (Boveri, 1914; 2008), aneuploidy is hall-mark of many cancers. However, whether aneuploidy is the cause or the result of cancer is still at debate. The molecular mechanism behind aneuploidy includes the chromosome mis-segregation in mitosis by the compromise of spindle assembly checkpoint (SAC). SAC is an elaborate network of proteins, which monitor that all chromosomes are bipolarly attached with the spindles. Therefore, the weakening of the SAC is the major reason for chromosome number instability, while complete compromise of SAC results in detrimental death, exemplified in natural abortion in embryonic stage. Here, I will review on the recent progress on the understanding of chromosome missegregation and cancer, based on the comparison of different mouse models of BubR1, the core component of SAC.