Sun Ginseng Protects Endothelial Progenitor Cells From Senescence Associated Apoptosis

Wooseok Im,Jin-Young Chung,Jaejun Bhan,Jiyeon Lim,Soon-Tae Lee,Kon Chu,Manho Kim 고려인삼학회 2012 Journal of Ginseng Research Vol.36 No.1

Endothelial progenitor cells (EPC) are a population of cells that circulate in the blood stream. They play a role in angiogenesis and, therefore, can be prognostic markers of vascular repair. Ginsenoside Rg3 prevents endothelial cell apoptosis through the inhibition of the mitochondrial caspase pathway. It also affects estrogen activity, which reduces EPC senescence. Sun ginseng (SG), which is heat-processed ginseng, has a high content of ginsenosides. The purpose of this study was to investigate the protective effects of SG on senescence-associated apoptosis in EPCs. In order to isolate EPCs, mononuclear cells of human blood buffy coats were cultured and characterized by their uptake of acetylated low-density lipoprotein (acLDL) and their binding of Ulex europaeus agglutinin I (ulex-lectin). Flow cytometry with annexin-V staining was performed in order to assess early and late apoptosis. Senescence was determined by β-galactosidase (β-gal) staining. Staining with 4’-6-Diamidino-2-phenylindole verifi ed that most adherent cells (93±2.7%) were acLDL-positive and ulex-lectin-positive. The percentage of β-gal-positive EPCs was decreased from 93.8±2.0% to 62.5±3.6% by SG treatment. A fl uorescence-activated cell sorter (FACS) analysis showed that 4.9% of EPCs were late apoptotic in controls. Sun ginseng decreased the apoptotic cell population by 39% in the late stage of apoptosis from control baseline levels. In conclusion, these results show antisenescent and antiapoptotic effects of SG in human-derived EPCs, indicating that SG can enhance EPC-mediated repair mechanisms.

Nuclear localization of huntingtin during spermatogenesis

Im, Wooseok,Chung, Jinyoung,Lee, Soon-Tae,Chu, Kon,Kim, Min-Wook,Kim, Manho Springer-Verlag 2014 Neurological sciences Vol.35 No.3

Applications of CRISPR/Cas9 for Gene Editing in Hereditary Movement Disorders

Im, Wooseok,Moon, Jangsup,Kim, Manho The Korean Movement Disorder Society 2016 Journal of movement disorders: official journal of Vol.9 No.3

<P>Gene therapy is a potential therapeutic strategy for treating hereditary movement disorders, including hereditary ataxia, dystonia, Huntington’s disease, and Parkinson’s disease. Genome editing is a type of genetic engineering in which DNA is inserted, deleted or replaced in the genome using modified nucleases. Recently, clustered regularly interspaced short palindromic repeat/CRISPR associated protein 9 (CRISPR/Cas9) has been used as an essential tool in biotechnology. Cas9 is an RNA-guided DNA endonuclease enzyme that was originally associated with the adaptive immune system of <I>Streptococcus pyogenes</I> and is now being utilized as a genome editing tool to induce double strand breaks in DNA. CRISPR/Cas9 has advantages in terms of clinical applicability over other genome editing technologies such as zinc-finger nucleases and transcription activator-like effector nucleases because of easy <I>in vivo</I> delivery. Here, we review and discuss the applicability of CRISPR/Cas9 to preclinical studies or gene therapy in hereditary movement disorders.</P>

Cytokine reporter mouse system for screening novel IL12/23 p40-inducing compounds.

Im, Wooseok,Kim, Hyojeong,Yun, Daesun,Seo, Sung-Yum,Park, Se-Ho,Locksley, Richard M,Hong, Seokmann Korean Society for Molecular Biology 2005 Molecules and cells Vol.20 No.2

<P>Cytokines interleukin (IL) 12 and 23 play critical roles in linking innate and adaptive immune responses. They are members of heterodimeric cytokines, sharing a subunit p40. Although IL12/23 p40 is mainly induced in macrophages and dendritic cells (DCs) after stimulation with microbial Toll-like receptor ligands, methods to monitor the cells that produce IL12/23 p40 in vivo are limited. Recently, the mouse model to track p40-expressing cells with fluorescent reporter, yellow fluorescent protein, has been developed. Macrophages and DCs from these mice faithfully reported p40 induction using the fluorescent marker. Here we took advantage of these reporter mice to screen bio-compounds for p40-inducing activity. After screening hundreds of compounds, we found several extracts inducing IL12/23 p40 gene expression. Treatment of DCs with these extracts induced the expression of MHC class II and co-stimulatory molecules, which implies that these might be useful as adjuvants. Next, the in vivo target immune cells of candidate compounds were examined. The reporter system can be useful to identify cells producing IL12 or IL23 in vivo as well as in vitro. Thus, our cytokine reporter system proved to be a valuable reagent for screening for immunostimulatory molecules and identification of target cells in vivo.</P>

Aberrant Expression Pattern and Location of Cullin 1 are Associated with the Development of Papillary Carcinoma in Thyroid and Cyclin D1 Expression

Wooseok Byon,Sung-Im Do,Kyungeun Kim,Hyunjoo Lee,Hyun-Soo Kim,Tae Gu Do,Jisup Yun,Dong-Hoon Kim,Seoung Wan Chae,Yong Lai Park,Chan Heun Park,Jin Hee Sohn 대한외과학회 2014 대한외과학회 학술대회 초록집 Vol.2014 No.5

초음파 속도법을 활용한 강판의 두께 변화 탐지를 위한 기초연구

김우석 ( Wooseok Kim ),문성모 ( Seongmo Mun ),김철민 ( Chulmin Kim ),임석빈 ( Seokbeen Im ) 한국구조물진단유지관리공학회 2020 한국구조물진단유지관리공학회 논문집 Vol.24 No.6

본 연구는 강교량의 폐합부재 등 시각적으로 탐지하기 어려운 부위의 강재에 대한 손상을 탐지하기 위한 방법을 개발하기 위한 목적의 기초연구이다. 여러 비파괴 방법들 중에서 초음파 속도법을 활용하여 두께가 다른 시편에 대해서 초음파 속도법을 실시하여, 평균 매질내전파속도를 도출하였고, 이를 활용하여 강재 부재에 대한 회귀분석을 실시하였다. 동일한 재료일 경우 회귀분석의 결과를 활용하면 다른 부재의 두께를 도출할 수 있을 것으로 예상된다. 또한, 광범위한 범위를 스캐닝하기 위해 200 mm/s로 이동하는 연속스캐닝 기법을 검토하였고, 두께가 변화는 부재의 두께를 효과적으로 예측할 수 있었다. This study was initiated to develop an effective inspection method to detect defects such as corrosion in closed-cell steel members in steel-box girder bridges. The ultrasonic velocity method among various non-destructive method was selected as a rapid and effective method to derive the average propagation velocity in the medium by using the ultrasonic wave velocity method for specimens of different thickness. The regression analysis was performed based on the experimental results, and the results was interpolated to evaluate the prediction accuracy. If the material properties are identical, this ultrasonic velocity method can predict the thickness using the averaged transmitted velocity. In addition, a continuous scanning method moving at 200 mm/s was tested for scanning a wide area of a bridge. The results exhibited that the continuous scanning method was able to effectively scan the different thickness of a bridge.

MicroRNA-124 slows down the progression of Huntington’s disease by promoting neurogenesis in the striatum

Liu, Tian,Im, Wooseok,Mook-Jung, Inhee,Kim, Manho Medknow PublicationsMedia Pvt Ltd 2015 Neural regeneration research Vol.10 No.5

<P>MicroRNA-124 contributes to neurogenesis through regulating its targets, but its expression both in the brain of Huntington’s disease mouse models and patients is decreased. However, the effects of microRNA-124 on the progression of Huntington’s disease have not been reported. Results from this study showed that microRNA-124 increased the latency to fall for each R6/2 Huntington’s disease transgenic mouse in the rotarod test. 5-Bromo-2’-deoxyuridine (BrdU) staining of the striatum shows an increase in neurogenesis. In addition, brain-derived neurotrophic factor and peroxisome proliferator-activated receptor gamma coactivator 1-alpha protein levels in the striatum were increased and SRY-related HMG box transcription factor 9 protein level was decreased. These findings suggest that microRNA-124 slows down the progression of Huntington’s disease possibly through its important role in neuronal differentiation and survival.</P>

Estimation of Density and Abundance of Euphausiids and Copepods using multi-frequency method

Kyounghoon Lee,Junghwa Choi,Wooseok Oh,Euna Yoon,Geunchang Park,Inwoo Han,Yangjae Im 한국수산해양기술학회(구 한국어업기술학회) 2018 한국수산해양기술학회 학술발표대회 Vol.2018 No.11

Exosome-Based Delivery of miR-124 in a Huntington’s Disease Model

Lee, Soon-Tae,Im, Wooseok,Ban, Jae-Jun,Lee, Mijung,Jung, Keun-Hwa,Lee, Sang Kun,Chu, Kon,Kim, Manho The Korean Movement Disorder Society 2017 Journal of movement disorders: official journal of Vol.10 No.1

<P><B>Objective </B></P><P>Huntington’s disease (HD) is a genetic neurodegenerative disease that is caused by abnormal CAG expansion. Altered microRNA (miRNA) expression also causes abnormal gene regulation in this neurodegenerative disease. The delivery of abnormally downregulated miRNAs might restore normal gene regulation and have a therapeutic effect. </P><P><B>Methods </B></P><P>We developed an exosome-based delivery method to treat this neurodegenerative disease. miR-124, one of the key miRNAs that is repressed in HD, was stably overexpressed in a stable cell line. Exosomes were then harvested from these cells using an optimized protocol. The exosomes (Exo-124) exhibited a high level of miR-124 expression and were taken up by recipient cells. </P><P><B>Results </B></P><P>When Exo-124 was injected into the striatum of R6/2 transgenic HD mice, expression of the target gene, RE1-Silencing Transcription Factor, was reduced. However, Exo-124 treatment did not produce significant behavioral improvement. </P><P><B>Conclusion </B></P><P>This study serves as a proof of concept for exosome-based delivery of miRNA in neurodegenerative diseases.</P>

사이토카인 리포터생쥐 시스템을 이용한 소수 인터루킨-12 p40 발현 면역세포의 확인

기미아(Mia Gi),김정은(Jungeun Kim),임우석(Wooseok Im),윤지희(Jeehee Youn),홍석만(Seokmann Hong) 대한체질인류학회 2009 해부·생물인류학 (Anat Biol Anthropol) Vol.22 No.1

인터루킨(IL)-12는 p35와 p40 단백질로 구성된 사이토카인으로, 선천면역과 적응면역을 서로 연결시켜주는 매우 중요한 면역학적 기능을 한다. p35 유전자는 지속적으로 발현되지만, p40 유전자의 발현은 외부자극에 의해 유도되는 점에 착안하여, 최근 IL-12 p40 유전자를 발현하는 세포를 형광 발현으로 동정할 수 있는 형광리포터생쥐 모델이 만들어졌다. 본 연구에서는 형광리포터 시스템을 이용하여 생체 내에 매우 적은 수로 존재하는 미세아교세포와 B 세포 소수 집단의 IL-12 p40 발현을 확인하고자 하였다. 먼저, IL-12 p40 형광리포터생쥐의 지라로부터 유래한 가지돌기세포와 복강으로부터 분리한 큰포식세포를 Toll-유사수용체(Toll-like receptor, TLR) 리간드로 자극한 후 IL-12 p40 발현 정도를 황색 형광단백질(yellow fluorescence protein, YFP) 발현으로 조사하였다. 그리고 뇌에 분포하는 큰포식세포의 한 종류인 미세아교세포를 신생 생쥐로부터 분리 증폭하여 IL-12 p40 발현을 조사하였다. 또한 지라의 B 세포에서 IL-12 발현이 유도되는지를 조사하였다. 지라 유래 가지돌기세포와 복강 유래 큰포식세포에서 IL-12 p40 형광리포터생쥐 시스템의 유용성을 확인하였다. 그리고 분리 증폭한 미세아교세포의 일부 세포가 TLR 리간드인 CpG oligodeoxynucleotides, lipopolysaccharide, peptidoglycan 그리고 polyinosinic-polycytidylic acid의 자극에 대해 IL-12 p40이 발현되는 것을 YFP 형광단백질 발현으로 확인할 수 있었다. 마지막으로 B 세포의 경우, B 세포의 소수 집단이 CpG 자극에 특이적으로 IL-12 p40을 발현하는 것을 확인하였다. 종합하면, 본 연구결과는 IL-12 p40 형광리포터생쥐 시스템을 통하여 뇌에 존재하는 미세아교세포의 일부와 소수의 B 세포집단에서 적절한 TLR 자극 시 IL-12 p40 유전자가 발현됨을 보였다. 이상의 결과는 IL-12 p40 형광리포터생쥐 시스템은 최근 새로이 발견되고 있는 소수 선천면역세포들을 효과적으로 동정하는 데 유용함을 제시한다. Interleukin-12 (IL-12), consisting of p35 and p40, plays important roles in linking innate and adaptive immunity. While p35 is constitutively expressed, IL-12 p40 gene expression is induced upon activation by Toll-like receptor ligands. Recently, with gene targeting technology, the cytokine IL-12 p40 reporter mouse has been developed to express the p40 gene linked via a viral IRES element with yellow fluorescence protein (YFP) fluorescent reporter. We investigated whether this novel system would be useful to reveal IL-12 p40-producing immune cells. We first investigated whether macrophages and dendritic cells from these mice faithfully reported p40 induction. Next, we tested if microglial cells, macrophages in the brain, could induce IL-12 p40. Finally we tested whether B cells could produce IL-12 p40 because there were very few reports for IL-12 production by B cells. Our results confirmed that macrophages and dendritic cells are main producer of IL-12 p40. Then, we found that microglial cells could produce IL-12 p40 upon stimulation with various TLR ligands. Finally we found that a subset of B cells could produce IL-12 p40 in TLR9-dependent manner. Taken all together, our system will be a valuable tool to identify the type of immune cells that produce IL-12 p40.