• Title/Summary/Keyword: gDNA

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The $Fc{\gamma}$ receptor III genotype as a risk factor for aggressive periodontitis in Korean patients (한국인 급진성 치주염 환자의 위험요소로서 $Fc{\gamma}$ III 수용기의 유전형)

  • Kim, Myung-Hyun;Shin, Seung-Yun;Kim, Tae-Il;Seol, Yang-Jo;Lee, Yong-Moo;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Ku, Young
    • Journal of Periodontal and Implant Science
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    • v.36 no.1
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    • pp.113-123
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    • 2006
  • 연구배경 IgG에 대한 $Fc{\gamma}$ 수용기는 치주병인균에 대한 숙주 반응에 있어서 중요한 역할을 하는데, 이 중 $Fc{\gamma}RIIIa$는 NK 세포, 대식세포, 단핵구, ${\gamma}{\delta}T$세포에서 발현되며, EC2 도메인에서 158 아미노산 부위의 valine (V)-phenylalanine (F)의 유전자다형성을 보인다. $Fc{\gamma}RIIIb$는 특이적으로 중성구에 발현되는데, extracellular (ECl) Ig-like 도메인 내 4개의 아미노산 치환(substitutions)에 의한 NA1-NA2 유전자다형성을 보인다. 이 연구의 목적은 한국인에서 급진성 치주염 환자와 $Fc{\gamma}III$ 수용기의 유전자다형성과의 관련성을 알아보는 것이다. 연구방법 및 재료 치주적으로 건강한 90명 (대조군, 남자 64명, 여자 26명)과 서울대학교 치과병원 지주과에 내원하여 급진성 치주염으로 진단된 환자 43명 (aggressive periodontitis patients: AgP, 남자 30명, 여자 13명)을 대상으로 하였다. 모든 실험 대상자는 임상 실험에 대해 동의 하였고, 초진 시 전자 탐침(Florida Probe(R) Co. Gainesville, FL)을 이용하여 탐침 시 치주낭 깊이 (PPD), 임상부착수준 (CAL), 치태지수(PI), 탐침 후 출혈지수 (BOP)를 측정하였다. 또한 이들의 정맥혈에서 추출한 DNA를 PCR법, 전기영동법 등을 이용하여 $Fc{\gamma}RIIIa$, $Fc{\gamma}RIIIb$의 대립 유전자의 존재여부를 확인하였다. 이를 바탕으로 $Fc{\gamma}RIII$ 복합 유전형을 확인하여 각 군 간을 비교하였다. 연구 결과 1. $Fc{\gamma}RIIIa$에 대한 유전자다형성 연구 결과 대조군과 급진성 치주염 환자 군(AgP)사이에서는 대립 유전자 분포가 서로 유의성 있는 차이를 나타내었고 (P<0.05), $Fc{\gamma}RIIIb$에서는 유의성 있는 차이를 발견할 수 없었다. (P>0.05) 2. $Fc{\gamma}RIIIa$ 158F 대립형질이 급진성 치주염 환자에서 유의성 있게 많이 발견되어졌다. (P<0.05) 결론 이 연구를 통하여 $Fc{\gamma}RIIIa$ 유전자의 분석이 한국인의 급진성 치주염에 대한 감수성의 위험요소의 표지자로 활용할 수 있을 것으로 사료되며, 향 후 더 많은 환자를 대상으로 히는 추가 연구가 필요하다고 생각된다.

Physiological Activities of Phellinus ribis Extracts (찔레 영지버섯(Phellinus ribis) 추출물의 생리활성)

  • Song, Jae-Hwan;Lee, Hyun-Sook;Hwang, Jin-Kook;Chung, Tae-Young;Hong, Sung-Ryul;Park, Ki-Moon
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.690-695
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    • 2003
  • Physiological activities of 40% ethanol extracts of Phellinus ribis were studied by employing several biological and biochemical assays. The extracts of Phellinus ribis displayed nitrate-scavenging activities (NSA) at pH 1.2 as with 64% NSA with 1.0 mg/mL of the extracts. They also had 91% 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities at the concentration of 1.0 mg/mL. Antioxidant activities of the extracts (at 0.5 mg/mL) on the autoxidation of linoleic acid (p<0.001) were also observed.. The inhibitory effect of the extracts on angiotensin converting enzyme was 11%. Cytotoxic effects of Phellinus ribis extracts against human cancer cell tines were also examined using MTT assay. The extracts (at 50 mg/mL) had severe growth inhibitory effects on A549, Hela, AGS, and SK-Hep-1, which were 8, 44, 76 and 42%, respectively. Ames test indicated that the extracts had no mutating effects on Salmonella typhimurium TA98 and TA100.

NADPH oxidase inhibitor diphenyleneiodonium induces p53 expression and cell cycle arrest in several cancer cell lines (NADPH oxidase 저해제인 diphenyleneiodonium의 p53 발현 및 암세포의 성장억제에 대한 연구)

  • Jo, Hong-Jae;Kim, Kang-Mi;Song, Ju-Dong;Park, Young-Chul
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.778-782
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    • 2007
  • The Diphenyleneiodonium (DPI) is widely used as an inhibitor of flavoenzymes, particularly NADPH oxidase. In this study, we investigated the effect of DPI on the cell growth progression of human colon cancer cells HCT-116 (wild-type p53), HT-29 (p53 mutant) and human breast cancer cells MCF-7 (wild-type p53). DPI treatment in cancer cells evoked a dose- and time-dependent growth inhibition, and also induced the cell cycle arrest in C2/M phase. The peak of cell population arrested in C2/M phase was observed at12 hr after treatment of DPI. In addition, DPI significantly induced the expression of p53, which induces proapoptotic genes in response to DNA damage or irreparable cell cycle arrest, at 6 hr in DPI-stimulated cells. However, a catechol apocynin, which inhibits the assembly of NADPH oxidase, did not induce p53 expression. This suggest that p53 expression induced by DPI is not associated with the inhibition of NADPH oxidase. In conclusion, we suggest that DPI induces the expression of wild-type p53 by ROS-in-dependent mechanism in several cancer cells, and upregulated p53 may be involved in regulatory mechanisms for growth inhibition and cell cycle arrest at C2/M phase in DPI-stimulated cells.

Characterization of a Novel DWD Protein that Participates in Heat Stress Response in Arabidopsis

  • Kim, Soon-Hee;Lee, Joon-Hyun;Seo, Kyoung-In;Ryu, Boyeong;Sung, Yongju;Chung, Taijoon;Deng, Xing Wang;Lee, Jae-Hoon
    • Molecules and Cells
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    • v.37 no.11
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    • pp.833-840
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    • 2014
  • Cullin4-RING ubiquitin ligase (CRL4) is a family of multi-subunit E3 ligases. To investigate the possible involvement of CRL4 in heat stress response, we screened T-DNA insertion mutants of putative CRL4 substrate receptors that exhibited altered patterns in response to heat stress. One of the mutants exhibited heat stress tolerance and was named heat stress tolerant DWD1 (htd1). Introduction of HTD1 gene into htd1-1 led to recovery of heat sensitivity to the wild type level, confirming that the decrease of HTD1 transcripts resulted in heat tolerance. Therefore, HTD1 plays a negative role in thermotolerance in Arabidopsis. Additionally, HTD1 directly interacted with DDB1a in yeast two-hybrid assays and associated with DDB1b in vivo, supporting that it could be a part of a CRL4 complex. Various heat-inducible genes such as HSP14.7, HSP21, At2g03020 and WRKY28 were hyper-induced in htd1-1, indicating that HTD1 could function as a negative regulator for the expression of such genes and that these genes might contribute to thermotolerance of htd1-1, at least in part. HTD1 was associated with HSP90-1, a crucial regulator of thermotolerance, in vivo, even though the decrease of HTD1 did not affect the accumulation pattern of HSP90-1 in Arabidopsis. These findings indicate that a negative role of HTD1 in thermotolerance might be achieved through its association with HSP90-1, possibly by disturbing the action of HSP90-1, not by the degradation of HSP90-1. This study will serve as an important step toward understanding of the functional connection between CRL4-mediated processes and plant heat stress signaling.

Surveillance of Acanthamoeba spp. and Naegleria fowleri in environmental water by using the duplex real-time PCR (Duplex real-time PCR을 이용한 수계 중 가시아메바와 파울러자유아메바 조사)

  • Kim, Min-jeong;Lee, Gyu-Cheol;Kim, Kunwoo;Lee, Hyunji;Kim, Min Young;Seo, Dae Keun;Lee, Jeong Yeob;Cho, Young-Cheol
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.98-104
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    • 2018
  • Naegleria fowleri and Acanthamoeba spp. are free-living amoebas that are widely distributed in natural environments. Although uncommon, infection with these protozoans can cause fatal disease in humans and animals. In this study, in order to select the appropriate method to survey Naegleria fowleri and Acanthamoeba spp. in water samples, four molecular biology techniques and one commercially available kit for real-time PCR were compared. The results indicated that the duplex real-time PCR was the most sensitive, and could be used to simultaneously detect two different free-living amoebas. Using the duplex real-time PCR approach, the two free-living amoebas were surveyed in three local streams in Daejeon, Republic of Korea. The concentrated free-living amoebas were inoculated onto non-nutrient agar plates which had been spread with heat-inactivated Escherichia coli and incubated for 5~7 days. After incubation, gDNA was extracted and used as the template for amplification by duplex real-time PCR. Acanthamoeba spp. and N. fowleri was detected from ten (83.3%) and two (16.6%) of the twelve samples, respectively. As these two free-living amoebas can be fatal, continuous surveillance is needed to track their distribution in the aquatic environment for the drinking water safety.

Effect of Superior Cervical Sympathetic Ganglion Block on Brain Injury Induced by Focal Cerebral Ischemia/Reperfusion in a Rat Model (상경부교감신경절블록이 백서의 국소 뇌허혈/재관류로 인한 뇌 손상에 미치는 영향)

  • Lee, Ae Ryoung;Yoon, Mi Ok;Kim, Hyun Hae;Choi, Jae Moon;Jeon, Hae Yuong;Shin, Jin Woo;Leem, Jeong Gill
    • The Korean Journal of Pain
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    • v.20 no.2
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    • pp.83-91
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    • 2007
  • Background: Cerebral blood vessels are innervated by sympathetic nerves that originate in the superior cervical ganglia (SCG). This study was conducted to determine the effect of an SCG block on brain injury caused by focal cerebral ischemia/reperfusion in a rat model. Methods: Male Sprague-Dawley rats (270-320 g) were randomly assigned to one of three groups (lidocaine, ropivacaine, and control). After brain injury induced by middle cerebral artery (MCA) occlusion/reperfusion, the animals were administered an SCG bloc that consisted of $30{\mu}l$ of 2% lidocaine or 0.75% ropivacaine, with the exception of animals in the control group, which received no treatment. Twenty four hours after brain injury was induced, neurologic scores were assessed and brain samples were collected. The infarct and edema ratios were measured, and DNA fragmented cells were counted in the frontoparietal cortex and the caudoputamen. Results: No significant differences in neurologic scores or edema ratios were observed among the three groups. However, the infarct ratio was significantly lower in the ropivacaine group than in the control group (P < 0.05), and the number of necrotic cells in the caudoputamen of the ropivacaine group was significantly lower than in the control group (P < 0.01). Additionally, the number of necrotic and apoptotic cells in theropivacaine group were significantly lower than inthe control group in both the caudoputamen and the frontoparietal cortex (P < 0.05). Conclusions: Brain injury induced by focal cerebral ischemia/reperfusion was reduced by an SCG block using local anesthetics. This finding suggests that a cervical sympathetic block could be considered as another treatment option for the treatment of cerebral vascular diseases.

Alkaliphilic Endoxylanase from Lignocellulolytic Microbial Consortium Metagenome for Biobleaching of Eucalyptus Pulp

  • Weerachavangkul, Chawannapak;Laothanachareon, Thanaporn;Boonyapakron, Katewadee;Wongwilaiwalin, Sarunyou;Nimchua, Thidarat;Eurwilaichitr, Lily;Pootanakit, Kusol;Igarashi, Yasuo;Champreda, Verawat
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1636-1643
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    • 2012
  • Enzymatic pre-bleaching by modification of pulp fibers with xylanases is an attractive approach to reduce the consumption of toxic bleaching chemicals in the paper industry. In this study, an alkaliphilic endoxylanase gene was isolated from metagenomic DNA of a structurally stable thermophilic lignocellulose-degrading microbial consortium using amplification with conserved glycosyl hydrolase family 10 primers and subsequent genome walking. The full-length xylanase showed 78% sequence identity to an endo-${\beta}$-1,4-xylanase of Clostridium phytofermentans and was expressed in a mature form with an N-terminal His6 tag fusion in Escherichia coli. The recombinant xylanase Xyn3F was thermotolerant and alkaliphilic, working optimally at $65-70^{\circ}C$ with an optimal pH at 9-10 and retaining >80% activity at pH 9, $60^{\circ}C$ for 1 h. Xyn3F showed a $V_{max}$ of 2,327 IU/mg and $K_m$ of 3.5 mg/ml on birchwood xylan. Pre-bleaching of industrial eucalyptus pulp with no prior pH adjustment (pH 9) using Xyn3F at 50 IU/g dried pulp led to 4.5-5.1% increase in final pulp brightness and 90.4-102.4% increase in whiteness after a single-step hypochlorite bleaching over the untreated pulp, which allowed at least 20% decrease in hypochlorite consumption to achieve the same final bleaching indices. The alkaliphilic xylanase is promising for application in an environmentally friendly bleaching step of kraft and soda pulps with no requirement for pH adjustment, leading to improved economic feasibility of the process.

중력이상을 이용한 한반도 모호면 추출에 관한 연구

  • 김정우;조진동;김원균;민경덕;황재하;이윤수;박찬홍;황종선
    • Proceedings of the KSEEG Conference
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    • 2003.04a
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    • pp.258-264
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    • 2003
  • 중력이상 및 수치고도모델을 이용하여 한반도 모호면 심도를 추출하였다 중력이상값은 인공위성고도레이더 관폭값을 주로 이용한 전지구 모델을 이용하여 데이터영역 뿐 만 아니라 주파수영역에서도 자료의 균질성을 확보하였다. 모호면 추출은 Kim et al. [2000a]에 의해 제안된 스펙트럼 대비법 및 후리에급수를 이용한 파워스펙트럼분석법을 이용하였다. 전자는 지각근형을 전제로, 지형에 의한 중력효과와 후리에어 중력이상을 파동수영역에서 대비하여 모호면의 심도를 계산하는 방법이고, 후자는 완전부우게 중력이상으로부터 푸리에변환을 이용하여 지하 밀도 변화층의 심도를 계산하는 방법이다. 이 두 모호면은 서로 0.53의 상관관계를 갖고 있으며, 이는 모호면 산출의 방법론적인 차이 및 계산상의 오차인 것으로 사료된다. 이렇게 두 가지 독립적인 방법으로 추출된 모호면을 하나로 통합하기 위한 한 방법으로, 두 모호면의 차이를 계산한 후, 이를 최소자승법을 이용, 두 모호면을 보정하였다. 결과적으로 한반도의 최종 모호면의 평균심도는 32.0km, 표준편차는 2.5km 이며, 최소, 최대 심도는 20.3, 36.6km으로 나타났다. 이 경우 지형에 의한 중력효과는 스펙트럼대비법에 의해 제거된 결과이나, 한반도의 지각이 완전한 지각판 내에 놓여 있어서 Airy-Heiskanin 지각균형설의 가정이 타당성이 있는가, 혹은 국부적인 응력장에 의해 한반도의 지각이 과연 얼마나 지지되고 있는가 하는 것에 대한 추가적인 연구가 필요하며, 이에 앞서, 일정한 밀도차를 갖는 연속적인 밀도변화층이 존재한다는 가정이 반드시 필요하다.에는 관련성을 갖고 있으며, 이는 유류 분해정도를 파악하는 지시자로써 특정 무기 오염물질을 이용할 수 있을 가능성이 있으므로 좀더 이들 관계성에 대한 연구가 진행될 필요성이 있다고 판단된다.고 과학적으로 분석할 수 있는 방법이 될 수 있을 것으로 기대된다. 의미를 되새기는 것으로 짧은 연구를 시작하겠다. 등은 활성 값이 70% 이상으로 퇴적물 독성이 상대적으로 낮았다. 이중나선 DNA 함량은 28.4 % - 49%로 대조군에 비해서 감소가 크다. 대부분의 정점이 대조군의 30% 내외로 정점 간의 차이는 크지는 않다. 그러나 다른 측정자료와 같이 정점 22에서 18%로 최소치를 나타내고, 정점 2, 12에서 20% 내외의 값을 보인다. 종합적으로 볼 때 오염물질의 유입이 크고, 광양제철 인근 정점 들이 모두 다른 정점에 비해서 낮아서, 퇴적물 독성이 높은 정점으로 조사되었다.hiwo의 광합성 능력은 낮은 농도들에서는 대조구와 유사하였으나, 5 $\mu\textrm{g}$/l의 높은 농도에서는 초기에 매우 낮은 광합성 능력을 보이다가 시간이 경과하면서 대조군보다 더 높은 경향을 나타냈다. 이러한 결과는 식물플랑크톤이 benso[a]pyrene의 낮은 농도에서 노출될 때는 이 물질을 탄소원으로 사용할 가능성이 있음을 시사한다. 본 연구의 결과들은 연안해역에 benso[a]pyrene과 같은 지속성 유기오염물질이 유입되었을 때 내정여부에 따라 식물플랑크톤 군집내 종 천이와 일차생산력에 크게 영향을 미칠 수 있음을 시사한다.TEX>5.2개)였으며, 등급별 회수율은 각각 GI(8.5%), GII

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The Effect of Pore Sizes on Poly(L-lactide-co-glycolide) Scaffolds for Annulus Fibrosus Tissue Regeneration (조직공학적 섬유륜재생을 위한 PLGA 지지체 제조시 다공크기의 영향에 관한 연구)

  • So, Jeong-Won;Jang, Ji-Wook;Kim, Soon-Hee;Choi, Jin-Hee;Rhee, John-M.;Min, Byung-Hyun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.6
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    • pp.516-522
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    • 2008
  • Biodegradable polymers have been used extensively as scaffolding materials to regenerate new tissues and the ingrowth of tissue have been reported to be dependent directly of the porosity, pore diameter, pore shape, and porous structure of the scaffold. In this study, porous poly (L-lactide-co-glycolide) (PLGA) scaffolds with five different pore sizes were fabricated to investigate the effect of pore sizes for AF tissue regeneration. Cellular viability and proliferation were assayed by MTT test. Hydroxyproline/DNA content of AF cells on each scaffold was measured. sGAG analyses were performed at each time point of 2 and 6 weeks. Scaffold seeded AF cells were implanted into the back of athymic nude mouse to observe the difference of formation of disc-like tissue depending on pore size in vivo. We confirmed that scaffold with $180{\sim}250{\mu}m$ pores displayed high cell viability in vitro and produced higher ECM than scaffold with other pore sizes in vivo.

Suicidal gene therapy with rabbit cytochrome P450 4B1/2-aminoanthracene or 4-ipomeanol system in human colon cancer cell

  • Jang, Su Jin;Kang, Joo Hyun;Moon, Byung Seok;Lee, Yong Jin;Kim, Kwang Il;Lee, Tae Sup;Choe, Jae Gol;Lim, Sang Moo
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.1 no.2
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    • pp.118-122
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    • 2015
  • Suicidal gene therapy is based on the transduction of tumor cells with "suicide" genes encoding for prodrug-activating enzymes that render target cells susceptible to prodrug treatment. Suicidal gene therapy results in the death of tumor with the expression of gene encoding enzyme that converts non-toxic prodrug into cytotoxic product. Cytochrome P450 4B1 (CYP4B1) activates 4-ipomeanol (4-IPO) or 2-aminoanthracene (2-AA) to cytotoxic furane epoxide and unsaturated dialdehyde intermediate.In this study, therapeutic effects of suicidal gene therapy with rabbit CYP4B1/2-AA or 4-IPO system were evaluated in HT-29 (human colon cancer cell). pcDNA-CYP4B1 vector was transfected into HT-29 by lipofection and stable transfectant was selected by treatment of hygromycin ($500{\mu}g/mL$) for 3 weeks. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed for confirmation of CYP4B1 expression in CYP4B1 gene transduced cell. The cytotoxic effects of CYP4B1 transduced cell were determined using dye-exclusion assay after treatment of 2-AA or 4-IPO for 96 hrs. Dye-exclusion assay showed that $IC_{50}$ of HT-29 and CYP4B1 transduced HT-29 was 0.01 mM and 0.003 mM after 4-IPO or 2-AA treatment at 96 hrs exposure, respectively. In conclusion, CYP4B1 based prodrug gene therapy probably have the potential for treatment of colorectal adenocarcinoma.