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Inhibitory Effect of Gallic acid on Production of Chemokine and Growth Factor in Mouse Macrophage Stimulated by Lipopolysaccharide (Gallic acid가 Lipopolysaccharide로 활성화된 마우스 대식세포의 케모카인과 성장인자 생성에 미치는 영향)

  • Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.4
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    • pp.586-591
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    • 2010
  • Chemokine and Growth Factor are major mediumtors of immuno-inflammatory pathway. The purpose of this study is to investigate whether productions of Chemokine and Growth Factor in lipopolysaccharide (LPS)-induced mouse macrophage RAW 264.7 cells are modulated by Gallic acid (GA), which is easily founded in tannin-containing natural materials such as red wine, green tea, grape juice, and Corni Fructus. Productions of Chemokine and Growth Factor were analyzed by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. At first, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, the antibody-conjugated beads were added and incubated for 30 minutes. After incubation, detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Based on fluorescence intensity, concentrations of Chemokine and Growth Factor were determined. The results of the experiment are as follows. GA significantly inhibited the production of interferon-inducible protein (IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor (VEGF) in LPS-induced RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). GA significantly inhibited the production of monocyte chemoattractant protein-1(MCP-1) and macrophage-colony stimulating factor(M-CSF) in LPS-induced RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). GA diminished the production of granulocyte macrophage-colony stimulating factor (GM-CSF) in LPS-induced RAW 264.7 cells. But GA did not show the inhibitory effect on the production of leukemia inhibitory factor (LIP) and macrophage inflammatory protein (MIP)-2 in LPS-induced RAW 264.7 cells. These results suggest that GA has the immuno-modulating activity related with its inhibitory effects on the production of IP-10, KC, MCP-1, VEGF, and M-CSF in LPS-induced macrophages.

Proteome Analysis of Mouse Adipose Tissue and Colon Tissue using a Novel Integrated Data Processing Pipeline

  • Park, Jong-Moon;Han, Na-Young;Kim, Hokeun;Hwang, Injae;Kim, Jae Bum;Hahm, Ki-Baik;Lee, Sang-Won;Lee, Hookeun
    • Mass Spectrometry Letters
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    • v.5 no.1
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    • pp.16-23
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    • 2014
  • Liquid chromatography based mass spectrometry (LC-MS) is a key technology for analyzing highly complex and dynamic proteome samples. With highly accurate and sensitive LC-MS analysis of complex proteome samples, efficient data processing is another critical issue to obtain more information from LC-MS data. A typical proteomic data processing starts with protein database search engine which assigns peptide sequences to MS/MS spectra and finds proteins. Although several search engines, such as SEQUEST and MASCOT, have been widely used, there is no unique standard way to interpret MS/MS spectra of peptides. Each search engine has pros and cons depending on types of mass spectrometers and physicochemical properties of peptides. In this study, we describe a novel data process pipeline which identifies more peptides and proteins by correcting precursor ion mass numbers and unifying multi search engines results. The pipeline utilizes two open-source software, iPE-MMR for mass number correction, and iProphet to combine several search results. The integrated pipeline identified 25% more proteins in mouse epididymal adipose tissue compared with the conventional method. Also the pipeline was validated using control and colitis induced colon tissue. The results of the present study shows that the integrated pipeline can efficiently identify increased number of proteins compared to the conventional method which can be a breakthrough in identification of a potential biomarker candidate.

Liquid chromatography-tandem mass spectrometric analysis of oleracone D and its application to pharmacokinetic study in mice

  • Lim, Dong Yu;Lee, Tae Yeon;Lee, Jaehyeok;Song, Im-Sook;Han, Young Taek;Choi, Min-Koo
    • Analytical Science and Technology
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    • v.34 no.5
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    • pp.193-201
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    • 2021
  • We have demonstrated a sensitive analytical method of measuring oleracone D in mouse plasma using a liquid chromatography-tandem mass spectrometry (LC-MS/MS). Oleracone D and oleracone F (internal standard) in mouse plasma samples were processed using a liquid-liquid extraction method with methyl tertbutyl ether, resulting in high and reproducible extraction recovery (80.19-82.49 %). No interfering peaks around the peak elution time of oleracone D and oleracone F were observed. The standard calibration curves for oleracone D ranged from 0.5 to 100 ng/mL and were linear with r2 of 0.992. The inter- and intra-day accuracy and precision and the stability fell within the acceptance criteria. The pharmacokinetics of oleracone D following intravenous and oral administration of oleracone D at doses of 5 mg/kg and 30 mg/kg, respectively, were investigated. When oleracone D was intravenously injected, it had first-order elimination kinetics with high clearance and volume of distribution values. The absolute oral bioavailability of this compound was calculated as 0.95 %, with multi-exponential kinetics. The low aqueous solubility and a high oral dose of oleracone D may explain the different elimination kinetics of oleracone D between intravenous and oral administration. Collectively, this newly developed sensitive LC-MS/MS method of oleracone D could be successfully utilized for investigating the pharmacokinetic properties of this compound and could be used in future studies for the lead optimization and biopharmaceutic investigation of oleracone D.

Technology Development for Non-Contact Interface of Multi-Region Classifier based on Context-Aware (상황 인식 기반 다중 영역 분류기 비접촉 인터페이스기술 개발)

  • Jin, Songguo;Rhee, Phill-Kyu
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.20 no.6
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    • pp.175-182
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    • 2020
  • The non-contact eye tracking is a nonintrusive human-computer interface providing hands-free communications for people with severe disabilities. Recently. it is expected to do an important role in non-contact systems due to the recent coronavirus COVID-19, etc. This paper proposes a novel approach for an eye mouse using an eye tracking method based on a context-aware based AdaBoost multi-region classifier and ASSL algorithm. The conventional AdaBoost algorithm, however, cannot provide sufficiently reliable performance in face tracking for eye cursor pointing estimation, because it cannot take advantage of the spatial context relations among facial features. Therefore, we propose the eye-region context based AdaBoost multiple classifier for the efficient non-contact gaze tracking and mouse implementation. The proposed method detects, tracks, and aggregates various eye features to evaluate the gaze and adjusts active and semi-supervised learning based on the on-screen cursor. The proposed system has been successfully employed in eye location, and it can also be used to detect and track eye features. This system controls the computer cursor along the user's gaze and it was postprocessing by applying Gaussian modeling to prevent shaking during the real-time tracking using Kalman filter. In this system, target objects were randomly generated and the eye tracking performance was analyzed according to the Fits law in real time. It is expected that the utilization of non-contact interfaces.

The Effect of Bacillus-Fermented Scutellariae Radix Acupuncture Solution on Interleukin Production in Mouse Macrophage Stimulated by Lipopolysaccharide (바실러스균 발효황금약침액이 Lipopolysaccharide로 활성화된 마우스 대식세포의 인터루킨 생성에 미치는 영향)

  • Park, Wan-Su
    • Korean Journal of Acupuncture
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    • v.27 no.2
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    • pp.95-105
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    • 2010
  • Objectives : The purpose of this study is to investigate the effect of Bacillus-fermented Scutellariae Radix acupuncture solution (SB) on interleukin(IL) production in mouse macrophage stimulatedby lipopolysaccaride(LPS). Methods : Productions of interleukins were measured y High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$(multi-analyte profiling beads) technology. To begin with, cell culture supernatant was obtained after treatment with LPS(1 ${\mu}g/mL$) and SB for 24 hour. Then, it was incubated with the antibody-conj${\mu}g$ated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. After incubating for 30 minutes, Strepavidin-conjugated Phycoerythrin(SAPE) was then added. Incubating for another 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Results : The results of the experiment are as follows. SB significantly inhibited the LPS-induced production of IL-3($9.15{\pm}0.35$ pg/mL) by $6.92{\pm}0.05,\;7.21{\pm}0.11,\;6.96{\pm}0.33,\;and\;7.45{\pm}0.74$ pg/mL at the concentration of 25, 50, 100, and 200 ${\mu}g/mL$ in mouse macrophage RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced production of IL-5($7.30{\pm}0.48$ pg/mL) by $6.50{\pm}0.29,\;6.30{\pm}0.25,\;6.30{\pm}0.25,\;and\;5.80{\pm}0.25$ pg/mL at the concentration of 25, 50 100, and 200 ${\mg}g/mL$ in RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced productiion of IL-9($17.26{\pm}0.19$ pg/mL) by $15.01{\pm}0.43$ pg/mL at the concentration of 25 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced productioh of IL-13($187.80{\pm}2.90$ pg/mL) by $152.80{\pm}4.25,\;172.80{\pm}3.97,\;162.10{\pm}6.67,\;and\;165.30{\pm}11.80$ pg/mL at the concentration fo 25, 50, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-17($18.30{\pm}0.95$ pg/mL) by $13.30{\pm}1.25,\;13.80{\pm}1.11,\;13.30{\pm}0.75,\;and\;14.00{\pm}1.08$ pg/mL at the concentration of 25, 50 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-23($43.90{\pm}0.83$ pg/mL by $39.50{\pm}1.26,\;38.00{\pm}1.78,\;and\;39.60{\pm}2.49$ pg/mL at the concentration of 25, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). Conclusions : These results suggest that SB has anti-inflammatory activity related with its inhibition of IL-3, IL-5, IL-13, IL-17, and IL-23 production in macrophages.

A $G_{4}$ Sequence within PHR1 Promoter Acts as a Gate for Cross-Talks between Damage-Signaling Pathway and Multi-Stress Response

  • Jang, Yeun-Kyu;Kim, Eun-Mi;Park, Sang-Dai
    • Animal cells and systems
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    • v.6 no.3
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    • pp.271-275
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    • 2002
  • Rph1 and Gisl are damage-responsive repressors involved in PHR1 expression. They have two $C_{2}$H/ sub 2/ zinc finger motifs as putative DNA binding domains and N-terminal conserved domain with unknown function. They are also found in the human retinoblastoma binding protein 2 and the mouse jumonji- encoded protein. The repressors are able to bind to A $G_{4}$ sequence within a 39-bp sequence called upstream repressing sequence of PHR1 promoter (UR $S_{PHR1}$) responsible for the damage-response of PHR1. We report here that Rph1 is predominantly localized in the nucleus as examined by fluorescence microscopic analysis with GFP-Rph1 fusion protein. On the basis of the fact that the A $G_{4}$ sequence that is recognized by Rph1 and Gisl is also recognized by Msn2 and Msn4 in a process of stress response, we a1so tried to examine the in vivo function of A $G_{4}$ and the role of Msn2 and Msn4 in PHR1 expression. Our results demonstrate that Msn2 and Msn4 are actually required for the basal transcription of PHR1 expression but not for its damage induction. When A $G_{4}$ sequence was inserted into the minimal promoter of the cyc1-LacZ reporter, the increased LacZ expression was observed indicating its involvement in transcriptional activation. The data suggest that the A $G_{4}$ is primarily required for basal transcriptional activation of PHR1 or CYC1 promoter through the possible involvement of Msn2 and Msn4. However, since the A $G_{4}$ is also involved in the repression of PHR1 via Rphl and Gisl, it is proposed that A $G_{4}$ functions as either URS or upstream activating sequence (UAS) depending on the promoter context.t.

Evaluation of Maternal Toxicity in Rats Exposed to Multi-Wall Carbon Nanotubes during Pregnancy

  • Lim, Jeong-Hyeon;Kim, Sung-Hwan;Lee, In-Chul;Moon, Chang-Jong;Kim, Sung-Ho;Shin, Dong-Ho;Kim, Hyoung-Chin;Kim, Jong-Choon
    • Environmental Analysis Health and Toxicology
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    • v.26
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    • pp.6.1-6.8
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    • 2011
  • Objectives: The present study investigated the potential adverse effects of multi-wall carbon nanotubes (MWCNTs) on pregnant dams and embryonic development following maternal exposure in rats. Methods: MWCNTs were orally administered to pregnant rats from gestational day (GD) 6 through 19 at dose levels of 0, 8, 40, 200, and 1000 mg/kg/day. During the test period, clinical signs, mortality, body weights, food consumption, serum biochemistry, oxidant-antioxidant status, gross findings, organ weights, and Caesarean section findings were examined. Results: All animals survived to the end of the study. A decrease in thymus weight was observed in the highest dose group. However, maternal body weight, food consumption, serum biochemical parameters, and oxidant-antioxidant balance in the kidneys were not affected by treatment with MWCNTs. No treatment-related differences in gestational index, embryo-fetal mortality, or fetal and placental weights were observed between treated and control groups. Conclusions: The results show that 14-day repeated oral dosing of MWCNTs during pregnancy induces minimal maternal toxicity at 1000 mg/kg/day in rats. Under these experimental conditions, the no-observed-adverse-effect level of MWCNTs is considered to be 200 mg/kg/day for dams and 1000 mg/kg/day for embryonic development.

The Design and Implement of Microarry Data Classification Model for Tumor Classification (종양 분류를 위한 마이크로어레이 데이터 분류 모델 설계와 구현)

  • Park, Su-Young;Jung, Chai-Yeoung
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.11 no.10
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    • pp.1924-1929
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    • 2007
  • Nowadays, a lot of related data obtained from these research could be given a new present meaning to accomplish the original purpose of the whole research as a human project. The method of tumor classification based on microarray could contribute to being accurate tumor classification by finding differently expressing gene pattern statistically according to a tumor type. Therefore, the process to select a closely related informative gene with a particular tumor classification to classify tumor using present microarray technology with effect is essential. In this thesis, we used cDNA microarrays of 3840 genes obtained from neuronal differentiation experiment of cortical stem cells on white mouse with cancer, constructed accurate tumor classification model by extracting informative gene list through normalization separately and then did performance estimation by analyzing and comparing each of the experiment results. Result classifying Multi-Perceptron classifier for selected genes using Pearson correlation coefficient represented the accuracy of 95.6%.

Development of Virtual Campus Information System using Interactive Virtual Reality Technology (상호작용 VR 기술을 이용한 가상 캠퍼스 안내 시스템 구현)

  • Kim, Jong-Nam;Na, Kil-Hang;Kim, Jong-Heon;Kim, Gyeong-Eop;Jung, Young-Kee
    • 한국HCI학회:학술대회논문집
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    • 2008.02a
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    • pp.779-784
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    • 2008
  • 가상현실 시스템에서 체험자가 현실과 같은 몰입을 느끼기 위해서는 하드웨어를 통한 가상환경 구축과 체험자와의 상호작용이 중요하다. 본 논문에서는 위치 추적 시스템(Motion Tracking System), Wand(3D Mouse) 및 HoloPoint 등 다양한 VR 인터페이스를 통해 체험자의 움직임, 위치, 동작을 인식하고 대형 멀티 디스플레이 시스템을 통해 입체영상을 제공하는 가상환경을 구축하고 체험자가 원하는 정보를 상호작용(Interaction)으로 제공할 수 있는 가상 캠퍼스 안내 시스템을 구현하고자 한다. 가상 캠퍼스 구축은 캠퍼스의 지형, 건물 및 구조물들의 정확한 형상을 얻기 위해 3D 스캐너를 이용하였고 획득된 데이터는 일련의 과정들을 거쳐 3D 모델로 생성된다. 이렇게 생성된 모델을 재배치 및 최적화하기 위해 모델링 소프트웨어를 사용하였다. 구축된 가상 캠퍼스와 위치 추적 시스템 및 Wand의 연동을 위해 VR 프로그래밍 하여 체험자의 움직임 및 동작을 콘텐츠에 그대로 적용시켰다. 여기에 키오스크 유형의 HoloPoint를 이용하여 체험자의 손동작으로 상호작용하는 안내시스템도 구축하였다. 상호작용 가능한 가상캠퍼스 안내 시스템은 가상현실 시스템 구축에 대한 또 다른 방법과 활용 예를 제시함으로써 가상전시관 및 가상체험관 등에 활용될 수 있을 것으로 기대된다.

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The Kleisin Subunits of Cohesin Are Involved in the Fate Determination of Embryonic Stem Cells

  • Koh, Young Eun;Choi, Eui-Hwan;Kim, Jung-Woong;Kim, Keun Pil
    • Molecules and Cells
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    • v.45 no.11
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    • pp.820-832
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    • 2022
  • As a potential candidate to generate an everlasting cell source to treat various diseases, embryonic stem cells are regarded as a promising therapeutic tool in the regenerative medicine field. Cohesin, a multi-functional complex that controls various cellular activities, plays roles not only in organizing chromosome dynamics but also in controlling transcriptional activities related to self-renewal and differentiation of stem cells. Here, we report a novel role of the α-kleisin subunits of cohesin (RAD21 and REC8) in the maintenance of the balance between these two stem-cell processes. By knocking down REC8, RAD21, or the non-kleisin cohesin subunit SMC3 in mouse embryonic stem cells, we show that reduction in cohesin level impairs their self-renewal. Interestingly, the transcriptomic analysis revealed that knocking down each cohesin subunit enables the differentiation of embryonic stem cells into specific lineages. Specifically, embryonic stem cells in which cohesin subunit RAD21 were knocked down differentiated into cells expressing neural alongside germline lineage markers. Thus, we conclude that cohesin appears to control the fate determination of embryonic stem cells.