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Busulfan Inhibits PCNA Expression but Induces The Expression of pRB

  • 천영신;주학진;권득남;김진희
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.21-21
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    • 2001
  • 일반적으로 세포는 방사능이나 항암제 등의 자극에 의해 DNA가 손상받았을 때 DNA를 합성하기 전, DNA변이를 복구하기 위해 cell cycle을 정지시키게 된다. pRB(retinoblatoma protein)는 이러한 cell cycle의 조절기작에서 중요한 역할을 담당하는 것으로 알려져 있다. G1기에서 S기로 진행하는 것을 조절하는 단백질인 pRB 은 E2F(cell cycle transcription factor)와 상호작용하여 cell cycle 진행에 필요한 전사활성을 억제, PCNA (proliferating cell nuclear antigen)의 합성을 저해한다. 또한, E2F와 결합된 pRB는 apoptosis를 제어하는 유전자를 조절하는 것으로 알려져 있다. Cell cycle에 영향을 미치는 항암제의 일종인 busulfan을 처리하면, 정소 내에 존재하는 대부분의 생식세포들은 사멸되고 spermatogonia만 남는 것으로 알려져 있다. 그러나 그 기작에 대해서는 자세히 연구된 바가 없다. 본 연구에서는 busulfan처리시 spermatogonial stem cell이 어떤 기작에 의해 손상받지 않고 유지되는지를 알아보고자 실험을 수행하였다. Busulfan을 처리한 마우스 (항암제 투여 후 5주)와 정상적인 13주령의 마우스의 정소로부터 각각 세포를 분리하였다. LSC (laser scanning cytometry)를 이용하여 처리군(busulfan treated mice)과 대조군(normal mature mice)에 대해 각각 DNA함량을 비교ㆍ분석한 결과 G0/G1(2N)에 머물러 있는 세포비율이 처리군에서 현저하게 증가했다 (79.3$\pm$5.5%:8.1$\pm$1.3%). Cell cycle의 G1/S check point인 pRB와 PCNA 발현을 Western blot과 면역조직학적인 방법(immunohisto-chemistry)을 이용하여 조사하였다 PCNA는 대조군과 비교해, 처리군에서 매우 낮은 수준으로 발현되었다. 면역염색된 정소단면을 살펴보면, 대조군에서는 모든 세정관에서 PCNA를 발현하는 세포가 높은 비율로 검출되었고, 처리군에서는 소수의 세정관에서 세포들이 낮은 수준으로 검출되었다. 반면에, pRB의 경우 PCNA와는 상반된 결과를 나타내어, 대조군에서는 거의 발현이 되지 않는 반면, 처리군에서는 대부분의 세정관내, 기저막을 따라 위치한 세포들에서 발현되었다. 이상의 결과는 busulfan에 의해 pRB의 인산화가 억제, pRB 와 결합된 E2F는 전사 활성이 억제되어, PCNA 합성을 저해하는 것으로 설명되어질 수 있다. 결론적으로, 인산화가 억제된 pRB (underphosphorylated RB protein)이 quiescent spermatogonial stem cell에서만 특이하게 발현하는 단백질이며, 이러한 pRB의 발현은 apoptosis를 제어하는 역할을 담당해 busulfan처리에 의해 손상받지 않고 남아있는 것으로 시사된다.

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Butyrate modulates bacterial adherence on LS174T human colorectal cells by stimulating mucin secretion and MAPK signaling pathway

  • Jung, Tae-Hwan;Park, Jeong Hyeon;Jeon, Woo-Min;Han, Kyoung-Sik
    • Nutrition Research and Practice
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    • v.9 no.4
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    • pp.343-349
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    • 2015
  • BACKGROUND/OBJECTIVES: Fermentation of dietary fiber results in production of various short chain fatty acids in the colon. In particular, butyrate is reported to regulate the physical and functional integrity of the normal colonic mucosa by altering mucin gene expression or the number of goblet cells. The objective of this study was to investigate whether butyrate modulates mucin secretion in LS174T human colorectal cells, thereby influencing the adhesion of probiotics such as Lactobacillus and Bifidobacterium strains and subsequently inhibiting pathogenic bacteria such as E. coli. In addition, possible signaling pathways involved in mucin gene regulation induced by butyrate treatment were also investigated. MATERIALS/METHODS: Mucin protein content assay and periodic acid-Schiff (PAS) staining were performed in LS174T cells treated with butyrate at various concentrations. Effects of butyrate on the ability of probiotics to adhere to LS174T cells and their competition with E. coli strains were examined. Real time polymerase chain reaction for mucin gene expression and Taqman array 96-well fast plate-based pathway analysis were performed on butyrate-treated LS174T cells. RESULTS: Treatment with butyrate resulted in a dose-dependent increase in mucin protein contents in LS174T cells with peak effects at 6 or 9 mM, which was further confirmed by PAS staining. Increase in mucin protein contents resulted in elevated adherence of probiotics, which subsequently reduced the adherent ability of E. coli. Treatment with butyrate also increased transcriptional levels of MUC3, MUC4, and MUC12, which was accompanied by higher gene expressions of signaling kinases and transcription factors involved in mitogen-activated protein kinase (MAPK) signaling pathways. CONCLUSIONS: Based on our results, butyrate is an effective regulator of modulation of mucin protein production at the transcriptional and translational levels, resulting in changes in the adherence of gut microflora. Butyrate potentially stimulates the MAPK signaling pathway in intestinal cells, which is positively correlated with gut defense.

The Effects of the Okbyeongpung-san Plus ocheongryong-tang on the Rat Model with Ovalbumin-induced Allergic Rhinitis (옥병풍산합소청룡탕(玉屛風散合小靑龍湯)이 알레르기 비염 모델 흰 쥐에 미치는 영향)

  • Kim, Suk-San;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.19 no.2
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    • pp.88-98
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    • 2006
  • Background : Allergic rhinitis is an inflammation of the nasal mucosa which is characterized by sneezing, coughing itchy nose, mouth and throat, congestion and/or nasal discharge. Object : We have studied effects of the Okbyeongpung-san plus Socheongryong-tang on the change of the amounts of IL-4, II-5, $IFN-{\gamma}$ and total IgE in rats OVA-induced allergic rhinitis. Method : The 15 rats were divided into three groups ; normal group, control group, and sample group. To induce allergic rhinitis in control group and sample group , rats were sensitized intraperitoneally with 0.1% ovalbumin(OVA) solution 3 times at intervals of 1 week. Then intranasal sensitization was performed by diffusing 0.1% ovalbumin(OVA) solution 3 times at intervals of 2 days. After that time, rats in the sample group were oral administration treated by Okbyeongpung-san plus Socheongryong-tang 28 days. We observed the change of the amounts of IL-4, II-5, $IFN-{\gamma}$ and total IgE in rats in each grout. Result : 1. In Total IgE study, the treated group was proved significant inhibitory effect(p<0.05) 2. In Interleukin-4(IL-4) study, the treated group was proved significant inhibitory effect(p<0.001> 3. In IL-5 study, the treated group was proved significant inhibitory effect(p<0.001> 4. In Interferone-${\gamma}(IFN-{\gamma})$ study, the treated group was proved significant inhibitory effect(p<0.005) Conclusion : According to the above results, it is considered that the Okbyeongpung-san flus Socheongyong-tang has inhibitory effects on the allergic rhinitis of rats.

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Temporal and Spatial Distribution of Particulate Carcinogens and Mutagens in Bangkok, Thailand

  • Pongpiachan, Siwatt;Choochuay, C.;Hattayanone, M.;Kositanont, C.
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1879-1887
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    • 2013
  • To investigate the level of genotoxicity over Bangkok atmosphere, $PM_{10}$ samples were collected at the Klongchan Housing Authority (KHA), Nonsree High School (NHS), Watsing High School (WHS), Electricity Generating Authority of Thailand (EGAT), Chokchai 4 Police Station (CPS), Dindaeng Housing Authority (DHA) and Badindecha High School (BHS). For all monitoring stations, each sample covered a period of 24 hours taken at a normal weekday every month from January-December 2006 forming a database of 84 individual air samples (i.e. $12{\times}7=84$). Atmospheric concentrations of low molecular weight PAHs (i.e. phenanthrene, anthracene, pyrene and fluoranthene) were measured in $PM_{10}$ at seven observatory sites operated by the pollution control department of Thailand (PCD). The mutagenicity of extracts of the samples was compared in Salmonella according to standard Ames test method. The dependence of the effects on sampling time and on sampling location was investigated with the aid of a calculation of mutagenic index (MI). This MI was used to estimate the increase in mutagenicity above background levels (i.e. negative control) at the seven monitoring sites in urban area of Bangkok due to anthropogenic emissions within that area. Applications of the AMES method showed that the average MI of $PM_{10}$ collected at all sampling sites were $1.37{\pm}0.10$ (TA98; +S9), $1.24{\pm}0.08$ (TA98; -S9), $1.45{\pm}0.10$ (TA100; +S9) and $1.30{\pm}0.09$ (TA100; -S9) with relatively less variations. Analytical results reconfirm that the particulate PAH concentrations measured at PCD air quality monitoring stations are moderately low in comparison with previous results observed in other countries. In addition, the concept of incremental lifetime particulate matter exposure (ILPE) was employed to investigate the potential risks of exposure to particulate PAHs in Bangkok atmosphere.

A Design of the SMBC for Improving Reliability of Blocking Spam Mail (스팸 메일 차단 신뢰도 향상을 위한 SMBC 플랫폼 설계)

  • Park Nho-Kyung;Han Sung-Ho;Seo Sang-Jin;Jin Hyun-Joon
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.30 no.11B
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    • pp.730-735
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    • 2005
  • While the E-mail is a important way of fast communication in these days. it is real that the E-mail is often misused as a commercial advertisement method and creates many social problems. Even though various filtering techniques for blocking spam mails have been developed, reliability of mail systems is decreased by misreading normal mails as spam mails, i.e. false-positive errors. In this paper, the SMBC(Spam Mail Blocking Center) platform employing spam mail recovery method based on privacy information is proposed and designed. The SMBC is designed in frame layer based on spam blocking system of proxy sewer and can be physically implemented in various topology so that flexible development with layered module is possible. Using privacy information makes the proposed SMBC platform minimize processing load and false-positive error rates so that it can improve mail system reliabilities.

The New Design Methodology Considering Transistor Layout Variation (트랜지스터 레이아웃 산포를 고려한 새로운 설계 기법)

  • Doh, Ji Seong;Cho, Jun Dong
    • Journal of the Institute of Electronics and Information Engineers
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    • v.49 no.12
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    • pp.234-241
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    • 2012
  • This paper proposes a novel design methodology considering transistor layout variation. The proposed design technique is to improve the transistor's electrical characteristics without performing a circuit simulation to extract transistor layout variation. There are three advantages in the proposed method. Firstly, there is no need to change the normal design flow used in layout designs. Secondly, there is no need to perform simulation in order to extract the transistor layout variation. Thirdly, early warnings in layout design lead to decreasing the number of post layout simulations. Less post layout simulations will decrease the number of iterations in the design cycle and shorten design period. The number of bad transistors in the early design phase were reduced from 17.8% to 2.9% by applying eDRC environment for layout designers to develop Standard Cell Library.

Meteorological Characteristics related to the Variation in Ozone Concentrations before, during, and after the Typhoon Period in the Korean Peninsula (한반도 태풍영향 전·중·후 시기동안 오존농도 변화에 관한 기상특성 분석)

  • Shin, Hyeonjin;Song, Sang-Keun
    • Journal of Environmental Science International
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    • v.26 no.5
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    • pp.621-638
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    • 2017
  • Meteorological characteristics related to variations in ozone ($O_3$) concentrations in the Korean peninsula before, during, and after Typhoon Talas (1112) were analyzed using both observation data and numerical modeling. This case study takes into account a high $O_3$ episode (e.g., a daily maximum of ${\geq}90ppb$) without rainfall. Before the typhoon period, high $O_3$ concentrations in the study areas (e.g., Daejeon, Daegu, and Busan) resulted from the combined effects of stable atmospheric conditions with high temperature under a migratory anticyclone (including subsiding air), and wind convergence due to a change in direction caused by the typhoon. The $O_3$ concentrations during the typhoon period decreased around the study area due to very weak photochemical activity under increased cloud cover and active vertical dispersion under a low pressure system. However, the maximum $O_3$ concentrations during this period were somewhat high (similar to those in the normal period extraneous to the typhoon), possibly because of the relatively slow photochemical loss of $O_3$ by a $H_2O+O(^1D)$ reaction resulting from the low air temperature and low relative humidity. The lowest $O_3$ concentrations during the typhoon period were relatively high compared to the period before and after the typhoon, mainly due to the transport effect resulting from the strong nocturnal winds caused by the typhoon. In addition, the $O_3$ increase observed at night in Daegu and Busan was primarily caused by local wind conditions (e.g., mountain winds) and atmospheric stagnation in the wind convergence zone around inland mountains and valleys.

Behavior Analysis of PPWS Sockets for Suspension Bridges Considering Frictional Contact (마찰 접촉을 고려한 현수교 PPWS용 소켓의 거동 해석)

  • Yoo, Hoon;Lee, Sung-Hyung;Seo, Ju-Won
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.33 no.4
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    • pp.1281-1293
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    • 2013
  • A sophisticated finite element model is illustrated to analyze the behavior of Prefabricated Parallel Wire Strand(PPWS) sockets for main cables of suspension bridges. An orthotropic model is proposed for the casting material by considering both effects of individual wires and a casting alloy, and the contact between surfaces of a socket and a casting alloy is idealized by using the Coulomb friction and the surface-based contact model. The proposed FE model is verified by comparing the strain distributions obtained from the tensile test and FE analysis. The mechanical behavior of a socket is investigated with respect to the variation of the frictional coefficient. The result shows that the friction between surfaces significantly diminishes the stress concentration of a socket and a casting alloy, and the normal stress from the design equation represents the averaged value of the upper and lower quartiles in the distribution of contact stresses between a socket and a casting alloy.

Identification and Characterization of Single Nucleotide Polymorphisms of SLC22A11 (hOAT4) in Korean Women Osteoporosis Patients

  • Lee, Woon Kyu;Kwak, Jin Oh;Hwang, Ji-Sun;Suh, Chang Kook;Cha, Seok Ho
    • Molecules and Cells
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    • v.25 no.2
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    • pp.265-271
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    • 2008
  • Single nucleotide polymorphisms (SNPs) are the most common form of human genetic variation. Non-synonymous SNPs (nsSNPs) change an amino acid. Organic anion transporters (OATs) play an important role in eliminating or reabsorbing endogenous and exogenous organic anionic compounds. Among OATs, hOAT4 mediates high affinity transport of estrone sulfate and dehydroepiandrosterone sulfate. The rapid bone loss that occurs in post-menopausal women is mainly due to a net decrease of estrogen. In the present study we searched for SNPs within the exon regions of hOAT4 in Korean women osteoporosis patients. Fifty healthy subjects and 50 subjects with osteoporosis were screened for genetic polymorphism in the coding region of SLC22A11 (hOAT4) using GC-clamp PCR and denaturing gradient gel electrophoresis (DGGE). We found three SNPs in the hOAT4 gene. Two were in the osteoporosis group (C483A and G832A) and one in the normal group (C847T). One of the SNPs, G832A, is an nsSNP that changes the $278^{th}$ amino acid from glutamic acid to lysine (E278K). Uptake of [$3^H$] estrone sulfate by oocytes injected with the hOAT4 E278K mutant was reduced compared with wild-type hOAT4. Km values for wild type and E278K were $0.7{\mu}M$ and $1.2{\mu}M$, and Vmax values were 1.8 and 0.47 pmol/oocyte/h, respectively. The present study demonstrates that hOAT4 variants can causing inter-individual variation in anionic drug uptake and, therefore, could be used as markers for certain diseases including osteoporosis.

Mentha arvensis Attenuates Cognitive and Memory Impairment in Scopolamine-treated Mice (Scopolamine 처리에 의한 인지 및 기억력 손상 마우스에서 박하의 효과)

  • Lee, Jihye;Kim, Hye-Jeong;Jang, Gwi Yeong;Seo, Kyung Hye;Kim, Mi Ryeo;Choi, Yun Hee;Jung, Ji Wook
    • Korean Journal of Pharmacognosy
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    • v.51 no.1
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    • pp.70-77
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    • 2020
  • Mentha arvensis is used traditional medicine to treat various disorders. In the present study, M. arvensis were extracted by the solid-phase microextraction (SPME) method and analyzed by gas chromatograph-mass spectrometry (GC-MS). We investigated the protective effects and mechanisms of a M. arvensis extract on scopolamine-induced cognitive and memory impairment. Mice were orally pretreated with a M. arvensis extract or normal saline, and then behavior tests were conducted 30 min after scopolamine injection. The antioxidant capacities were analyzed by free radical scavenging (DPPH and ABTS). Acetylcholinesterase (AChE) activity were also measured using Ellman's method ex vivo test. In behavior tests, percent of spontaneous alteration, escape latency and swimming time in target quadrant were improved by the administration of the M. arvensis extract, which suggests that the M. arvensis extract improves memory function in the scopolamine-treated mice model. In addition, M. arvensis extract showed inhibition of the free radical and AChE activity. The results of the present study suggest that the M. arvensis extract ameliorates scopolamine-induced cognitive and memory deficits through the inhibition of free radicals and AChE activity. Therefore, M. arvensis may be a promising neuroprotective agent for management of learning and memory improvements in human dementia patients.