• Parasites, Hosts and Diseases
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• 제54권1호
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• pp.39-46
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• 2016

Theileria annulata is a tick-borne intracellular protozoan parasite that causes tropical theileriosis, a fatal bovine lymphoproliferative disease. The parasite predominantly invades bovine B lymphocytes and macrophages and induces host cell transformation by a mechanism that is not fully comprehended. Analysis of signaling pathways by quantitative real-time PCR (qPCR) could be a highly efficient means to understand this transformation mechanism. However, accurate analysis of qPCR data relies on selection of appropriate reference genes for normalization, yet few papers on T. annulata contain evidence of reference gene validation. We therefore used the geNorm and NormFinder programs to evaluate the stability of 5 candidate reference genes; 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ACTB (${\beta}-actin$), PRKG1 (protein kinase cGMP-dependent, type I) and TATA box binding protein (TBP). The results showed that 18S rRNA was the reference gene most stably expressed in bovine PBMCs transformed and non-transformed with T. annulata, followed by GAPDH and TBP. While 18S rRNA and GAPDH were the best combination, these 2 genes were chosen as references to study signaling pathways involved in the transformation mechanism of T. annulata.

• The Korean Journal of Physiology and Pharmacology
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• 제20권3호
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• pp.315-324
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• 2016

Human cardiac fibroblasts (HCFs) have various voltage-dependent $K^+$ channels (VDKCs) that can induce apoptosis. Hydrogen peroxide ($H_2O_2$) modulates VDKCs and induces oxidative stress, which is the main contributor to cardiac injury and cardiac remodeling. We investigated whether $H_2O_2$ could modulate VDKCs in HCFs and induce cell injury through this process. In whole-cell mode patch-clamp recordings, application of $H_2O_2$ stimulated $Ca^{2+}-activated$ $K^+$ ($K_{Ca}$) currents but not delayed rectifier $K^+$ or transient outward $K^+$ currents, all of which are VDKCs. $H_2O_2-stimulated$ $K_{Ca}$ currents were blocked by iberiotoxin (IbTX, a large conductance $K_{Ca}$ blocker). The $H_2O_2-stimulating$ effect on large-conductance $K_{Ca}$ ($BK_{Ca}$) currents was also blocked by KT5823 (a protein kinase G inhibitor) and 1 H-[1, 2, 4] oxadiazolo-[4, 3-a] quinoxalin-1-one (ODQ, a soluble guanylate cyclase inhibitor). In addition, 8-bromo-cyclic guanosine 3', 5'-monophosphate (8-Br-cGMP) stimulated $BK_{Ca}$ currents. In contrast, KT5720 and H-89 (protein kinase A inhibitors) did not block the $H_2O_2-stimulating$ effect on $BK_{Ca}$ currents. Using RT-PCR and western blot analysis, three subtypes of $K_{Ca}$ channels were detected in HCFs: $BK_{Ca}$ channels, small-conductance $K_{Ca}$ ($SK_{Ca}$) channels, and intermediate-conductance $K_{Ca}$ ($IK_{Ca}$) channels. In the annexin V/propidium iodide assay, apoptotic changes in HCFs increased in response to $H_2O_2$, but IbTX decreased $H_2O_2$-induced apoptosis. These data suggest that among the VDKCs of HCFs, $H_2O_2$ only enhances $BK_{Ca}$ currents through the protein kinase G pathway but not the protein kinase A pathway, and is involved in cell injury through $BK_{Ca}$ channels.

• 약학회지
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• 제56권3호
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• pp.191-197
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• 2012

We previously reported that the novel isoflavone-free peptide mixture (black soybean peptide, BSP) had several beneficial effects like antiobesity and hypotriglyceridemic effect. However, there are no reports for BSP on anti-hypertensive activity. BSP induced heme oxygenase-1 (HO-1) in HUVECs, thus investigated the HO-1-induced activity in HUVECs and the anti-hypertensive effects in SHR animal model. BSP significantly induced HO-1 expression both at transcriptional and protein levels in a time- and dose-dependent manner as measured by RT-PCR and Western blot analysis, respectively. These inductions were abolished by pretreatment of N-acetyl-cystein (NAC, 1~10 mM), but not by employing Tempol, a superoxide dismutase (SOD) mimetic (1~5 mM). As expected, enzymatic activity (~2 fold) determined by bilirubin formation assay and cGMP concentration (~6 fold) were significantly increased in BSP-treated cells. Based on the numerous evidences on the beneficial effects of HO-1 and our results, we investigated in vivo effects of BSP on the antihypertensive activity. The administration of BSP (1% in drinking water) significantly decreased mean blood pressure (BP) (from $218.6{\pm}6.99$ to $190.0{\pm}3.42$ mm Hg, p<0.01). This result indicates that BSP is strong inducer of HO-1 expression, which may be triggered by oxidative stress, and has anti-hypertensive activity.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2006년도 총회 및 춘계학술발표회
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• pp.76-81
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• 2006

창녕군 증산리 지역은 신규 취수원 확보 일환으로 강변여과 방식 취수 가능성이 한국수자원공사에 의해서 평가되고 있는 지역이다. 강변여과 방식 취수의 타당성 평가를 위해 현장 지질조사, 시추조사, 전기 비저항 탐사 및 고해상도 탄성파 탐사를 수행하였다. 또한 대상 연구지역에서의 개략적 취수 가능량 산정을 위해 모델링 평가가 수행되었으며, 개별 정호에 대한 양수량 산정을 위해 시험 정호를 설치하였다. 현장 시추 조사와 시료에 대한 입도 분석 결과, 연구 지역의 충적층 두께는 35m 전후이며, 주 대수층 구간은 지표하 $25{\sim}35m$인 것으로 추정되었다. 또한 주 대수층 구간의 수리전도도는 $10^{-2}cm/sec$ 이상으로 주로 모래섞인 자갈층으로 구성되어있는 것으로 평가되었다. 또한 전기비저항 조사 결과는 부분적으로 매우 낮은 비저항 분포 지층을 보여주고 있으며, 이는 주로 실트 및 점토로 된 지층이 부분적으로 퇴적되어 있음을 지시하며, 고해상도 탄성파 탐사 결과는 전반적으로 지하수위는 지표하부 5m 전후에 분포하고, 충적층의 하부 경계는 35 내지 45m인 것으로 해석되었다. 지하수 모델링을 통해, 취수 목표량인 180,000톤/일은 주대수층까지의 지하수위 강하 없이 확보 가능할 것으로 평가되었다. 또한 개별 정호의 산출 특성을 평가하기 위해 시험 정호를 설치하여, 실제 2,700톤/일 예비 양수 시험을 수행하였다. 예비 양수 시험 결과, 양수정에서의 수위 강하는 개략 10m, 양수정관측정에서는 약 0.3m의 수위강하만이 관찰되었으며, 양수 영향권이 수 십 m를 넘지 않을 것으로 판단되었다.서의 S97과 JBR의 세포감염 억제율은 3.85%와 3.63%로 나타났다. $textsc{k}$-casein, CU는 로타바이러스 S97과 JBR에 대해 농도 2000UM에서 97%이상의 억제효과를 나타냈으며, sialic acid는 억제효과가 거의 없었다. K-casein, GMP는 송아지뿐만 아니라 유아의 로타바이러스에 의한 설사를 억제할 수 있을 것으로 기대된다.을 향상하기 위해서는 이러한 부위에 대한 미생물 오염을 낮출 수 있는 세심한 현장 품질관리가 필요하다.en and adolescents, analysed by country, age group and gender. The paper discusses the places young consumers can turn to in trying to fulfil their growing consumer needs. It also examines how much money is at their disposal. It then concludes by considering the influence of "financial socialization" on how young people deal with money.nsumption visions based on the various perspectives, consumers are influenced in the apparel buying decision-making. Many subjects reported experiencing positive affect when imagining positive outcomes of produc

• 한국동물학회지
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• 제27권2호
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• pp.103-116
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• 1984

유전자 구조 및 유전정보 흐름의 차이로 인하여 고등생물의 유전자를 미생물에 직접 cloning하면 원하는 유전자 산물을 얻지 못하는 경우가 많다. 이것을 극복하기 위해서는 화학적인 방법으로 유전자를 합성하든지, 또는 역제효소를 사용하여 고등생물의 mRNA로부터 유전자를 합성하여 cloning하는 방법을 사용한다. 본 연구에서는 oligo(dT)-cellulose column 방법으로 순수분리한 plasmid pBR322의 Pst I site에 cloning하였다. 우선 AMV reverse transcriptase로 primary cDNA를 합성하고, 알칼리를 처리하여 주형 RNA를 제거했다. 이번에는 이 primary cDNA를 주형으로 Klenow enzyme과 reverse transcriptase를 차례로 처리하여 double stranded DNA를 합성하고, 이 때 5' end 근처에 형성되는 hairpin loop을 Sl nuclease로 제거했다. Terminal deoxynucleotidyl transferase를 사용하여, 합성된 dsDNA에는 poly(dC) track을, Pst I endonuclease를 처리한 plasmid DNA에서는 poly(dG) track을 각각 붙인다음 이들을 서로 annealing시키고 E. coli에 transformation시켜서 크기가 큰 plasmid를 갖는 clone을 cracking 방법으로 일처 선별하였다. 이렇게 선별된 clone을 in 냐셔 hybridization 방법으로 조사하여 globin DNA가 들어간 colony를 이차 선별하고 여러 restriction enzyme으로 잘라보아 globin DNA가 cloning된 것을 확인하였다. 토끼 hemoglobin으로 immunize한 rat (Wistar)에서 뽑은 제일차 혈청과 염소에서 뽑은 제이차 혈청의 antibody를 사용한 radioimmunoassay방법으로, cloning된 globin gene이 대장균내에서 발현되는 지의 여부를 살펴 보았는데, 박테리아의 $\\beta$-lactamase와 토끼의 globin이 결합된 chimeric protein이 대장균 내에서 다량 합성되며, 이 단백질은 토끼 hemoglobin의 antigenic determinant를 가지고 있음을 알 수 있었다.

• 대한약침학회지
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• 제18권1호
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• pp.63-71
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• 2015

Objectives: This study was carried out to analyze the single-dose toxicity of Eun-Bi San pharmacopuncture injected into the muscle of Sprague-Dawley (SD) rats. Methods: All experiments were performed at Biotoxtech, an institution certified to conduct non-clinical studies under the Good Laboratory Practice (GLP) regulations. Six week old SD rats reared by ORIENTBIO were chosen for this pilot study. The reason SD rats were chosen is that they have been widely used in safety tests in the field of medicine, so the results can be easily compared with many other databases. The Eun-Bi San pharmacopuncture was made in a clean room at the Korean Pharmacopuncture Institute (KPI, K-GMP). The constituents of the Eun-Bi San pharmacopuncture are Angelicae gigantis radix, Strychni semen and Glycyrrhizae radix. These were extracted at low temperature and low pressure in an aseptic room at the KPI. Doses of Eun-Bi San pharmacopuncture, 0.25, 0.5 and 1.0 mL, were administered to the experimental groups, and a dose of normal saline solution, 1.0 mL, was administered to the control group. This study was performed under the approval of the Institutional Animal Ethics Committee of Biotoxtech Co., Ltd. Results: No deaths or abnormalities occurred in any of the four groups. No significant changes in weight, hematological parameters or clinical chemistry between the control group and the experimental groups were observed. To determine if abnormalities existed in any organs and tissues, we used microscopy to examine representative histological sections of each specified organ; the results showed no significant differences in any of the organs or tissues. Conclusion: The results of this study suggest that treatment with Eun-Bi San pharmacopuncture is relatively safe and that its clinical use may be beneficial. Further evaluations and studies on this subject will be needed to provide more concrete evidence in support of these conclusions.

• 대한자원환경지질학회:학술대회논문집
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• 대한자원환경지질학회 2002년도 추계 공동 심포지엄 논문집
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• pp.53-67
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• 2002

As in the past, we are concerned today with the magnitudes of mineral resources and the adequacy of these resources to meet future needs. In looking at global resource issues, we should consider the need for the resource, its supply, and the environmental consequences of using it. The need for a resource can become a resource dependency, specially as the global population expands and each of us becomes Increasingly dependent upon hundreds of natural materials. Therefore, our great mineral consumption makes the human population a true “Geologic Force”, which will be even more significant in the future when the global population is projected to reach alarming proportions. Although our supplies of mineral resources probably will be sufficient for the 21st century, the uneven distribution of minerals in the Earth's crust almost certainly will continue to be a major problem. The most likely result will be major shifts in both prices and sources of supply of many mineral resources. As for energy resources, we must avoid an obsessive dependency on one fuel and expand instead to other energy resources. Finally, because the use of resources affects the environment, we need to focus on resource exploitation and global pollution, particularly in regard to ground water and arable land. We must manage our resources so as to be in balance with our environment. And the accelerated industrialization of South Korean economy over the last three decades has resulted in the mass consumption of mineral commodities. South Korea has around 50 useful mineral commodities for the mineral industry, among 330 kinds of minerals described. The component ratio of the mining industry sector of the gross national production(GNP) in South Korea dropped from 1.2% in 1971 to 0.34% in 1997 due to the rapid growth of other industries in the country. During the period from 1971 to 1997, the average growth rate of mineral consumption in South Korea was 9.13% yearly and that of GMP per capita was 14.97%. The mineral consumptions per capita showed a continual increase during the last 30 years as follows(parenthesis: GW per capita); 0.99 metric tons in 1997($289), 3.83 metric tons in 1989($5, 210), 6.11 metric tons in 1995 ($10, 037), and 6.66 metric tons in 1997($9, 511). The total amount of mineral consumption in South Korea was 33 million tons of 32 mineral commodities in 1971, and 306 million metric tons of 47 mineral commodities in 1997.

• Journal of Acupuncture Research
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• 제19권5호
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• pp.92-111
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• 2002

Objective : Carthami Flos has been used as a herb to promote blood circulation to remove blood stasis in oriental medicine for many centuries, and Amun(GV15) has been used as a meridian point to treat apoplexy etc. To investigate treatment of cerevral vascular disease(CVA) by promoting blood circulation and removing blood stasis(活血化瘀法), we observed the experimental effects and mechanism of auqa-acupunture of Carthami Flos(ACF) injected into GV15 on cerevral hemodynamics and cardiovascular system of rats. Method : Aqua-acupuncture of Carthami Flos(ACF) was injected into GV15, and then we investigated experimental effects and mechanism of ACF on the cerebral hemodynamics[regional cerebral blood flow(rCBF), pial arterial diameter(PAD), meal arterial blood pressure(MABP)] and cardiovascular system[cardiac muscle contractile force(CMF), heart rate(HR)I by pretreatment with methylene blue(MTB) and indomethacin(IDN). The changes in rCBF, MABP, CMF and HR were tested by Laser Doppler Flowmetry(LDF), and the changes in PAD was determinated by video microscopy methods and video analyzer. Results :The results were as follows in normal rats ; The changes of rCBF and PAD were significantly increased by ACF($120{\mu}{\ell}/kg$) in a injected time-dependent manner, but MABP was not changed by ACF. The changes of cardiovascular system were increased by ACF in a injected time-dependent manner. And pretreatment with MTB was significantly inhibited ACE induced increase of rCBF and PAD, and was decreased ACF induced increase of HR. And pretreatment with IDN was increased ACF induced MABP and CMF. And the results were as follows in cerebral ischemic rats ; The changes of rCBF was increased stabilizly by treatment with ACF($120{\mu}{\ell}/kg$) in during the period of cerebral reperfusion, but pretreatment with MTB was increased ACF induced increase of rCBF during the period of cerebral reperfusion. The results were as follows in normal rats ; The changes of rCBF and PAD were significantly increased by ACF($120{\mu}{\ell}/kg$) in a injected time-dependent manner, but MABP was not changed by ACF. The changes of cardiovascular system were increased by ACF in a injected time-dependent manner. And pretreatment with MTB was significantly inhibited ACF induced increase of rCBF and PAD, and was decreased ACF induced increase of HR. And pretreatment with IDN was increased ACF induced MABP and CMF. And the results were as follows in cerebral ischemic rats ; The changes of rCBF was increased stabilizly by treatment with ACF($120{\mu}{\ell}/kg$) in during the period of cerebral reperfusion, but pretreatment with MTB was increased ACF induced increase of rCBF during the period of cerebral reperfusion Conclusions : In conclusion, ACF causes a diverse response of rCBF, PAD an HR, and action of ACF is mediated by cyclic GMP. I suggested that ACF has an anti-ischemic effect through the improvement of crebral hemodynamics in a transient cerebral ischemia.