• Parasites, Hosts and Diseases
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• 제55권5호
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• pp.491-503
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• 2017

The effects of tyrosine kinase inhibitors (TKIs) were evaluated on growth inhibition of intracellular Toxoplasma gondii in host ARPE-19 cells. The number of tachyzoites per parasitophorous vacuolar membrane (PVM) was counted after treatment with TKIs. T. gondii protein expression was assessed by western blot. Immunofluorescence assay was performed using Programmed Cell Death 4 (PDCD4) and T. gondii GRA3 antibodies. The TKIs were divided into 3 groups; non-epidermal growth factor receptor (non-EGFR), anti-human EGFR 2 (anti-HER2), and anti-HER2/4 TKIs, respectively. Group I TKIs (nintedanib, AZD9291, and sunitinib) were unable to inhibit proliferation without destroying host cells. Group II TKIs (lapatinib, gefitinib, erlotinib, and AG1478) inhibited proliferation up to 98% equivalent to control pyrimethamine ($5{\mu}M$) at $20{\mu}M$ and higher, without affecting host cells. Group III TKIs (neratinib, dacomitinib, afatinib, and pelitinib) inhibited proliferation up to 98% equivalent to pyrimethamine at $1-5{\mu}M$, but host cells were destroyed at $10-20{\mu}M$. In Group I, TgHSP90 and SAG1 inhibitions were weak, and GRA3 expression was moderately inhibited. In Group II, TgHSP90 and SAG1 expressions seemed to be slightly enhanced, while GRA3 showed none to mild inhibition; however, AG1478 inhibited all proteins moderately. Protein expression was blocked in Group III, comparable to pyrimethamine. PDCD4 and GRA3 were well localized inside the nuclei in Group I, mildly disrupted in Group II, and were completely disrupted in Group III. This study suggests the possibility of a vital T. gondii TK having potential HER2/4 properties, thus anti-HER2/4 TKIs may inhibit intracellular parasite proliferation with minimal adverse effects on host cells.

• 산업융합연구
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• 제20권1호
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• pp.23-33
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• 2022

본 연구는 제조기업의 디지털 전환(DX)을 위한 실행모형을 개발하기 위해 진행하였다. 이를 위하여 경영혁신과 디지털전환 과정에 대한 선행연구를 검토하였다. 인터넷 비즈니스의 혁신과정에서 적용된 NEBIC이론과 혁신이론을 기반으로 DX과정모형을 도출하였다. 여기에 최고경영층(TMT)의 추진의지 요인을 포함한 연구모형을 구축하고 이를 검증하기 위해 험자료를 통해 확인하였다. 디지털 전환을 추진하고 있는 중견 제조기업의 구성원을 대상으로 수집된 자료를 기반으로 연구가설을 검증하였다. 회귀분석을 통해 연구모형(기술지식 → 기회인지 → 기대성과 → 실행의지)의 각 단계의 영향관계를 확인하였다. DX과정에 최고경영층의 DX추진의지에 대한 구성원의 인식의 매개효과를 파악하기 위해 위계적 회귀분석을 실시였다. Sobel test 확인결과, 각 단계별 관계에 경영층의 DX추진에 대한 인식이 부분적으로 매개하는 사실을 확인하였다. 본 연구는 중견 제조업의 디지털 전환에 적용 가능한 모형을 제시했다는 점에서 의미가 있다. 또한 혁신연구 및 NEBIC 확장에 대한 경험적 근거를 제공한 것에 가치를 지닌다. 동적특성을 지닌 과정모형에 대한 실증자료의 한계를 극복하기 위해서 종단연구가 필요하며 연구결과의 일반화를 위해 제조업 이외 다양한 분야에서 확장된 실증연구가 요구된다.

• Molecular & Cellular Toxicology
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• 제1권3호
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• pp.172-178
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• 2005

[ $21{\alpha}$ ]- and ${\beta}$-Methylmelianodiol were isolated as the inhibitor of IL-5 bioactivity from Poncirus tripoliata. To develope as an anti-septic drug, the genotoxicity of $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ was subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), single cell gel electrophoresis (Comet) assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. These compounds are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. Before performing the comet assay, $IC_{20}$ of $21{\alpha}-methylmelianodiol$ was determined the concentration of $25.51\;{\mu}g/mL\;and\;21.99\;{\mu}g/mL$ with and without S-9, respectively. Also $21{\beta}-methylmelianodiol$ was determined the concentration of $24.15\;{\mu}g/mL\;and\;\;22.46\;{\mu}g/mL$ with and without S-9, respectively. In the comet assay, DNA damage was not observed both $21{\alpha}-methylmelianodiol\;and\;21{\beta}-methylmelianodiol$ in mouse lymphoma cell line. Also, the mutant frequencies in the treated cultures were similar to the vehicle controls, and none of $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ with and without S-9 doses induced a mutant frequency over. twice the background. It is suggests that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ are non-mutagenic in MOLY assay. The results of this battery of assays indicate that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ have no genotoxic potential in bacterial or mammalian cell systems. Therefore, we suggest that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$, as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen.

• Asian-Australasian Journal of Animal Sciences
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• 제30권2호
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• pp.246-253
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• 2017

Objective: Present study explores the effect of hot summer period on the glycolytic rate of early post-mortem meat quality of Ghungroo and Large White Yorkshire (LWY) pig and comparative adaptability to high temperature between above breeds by shifting the expression of stress related genes like mono-carboxylate transporters (MCTs) and heat shock proteins (HSPs). Methods: Healthy pigs of two different breeds, viz., LYW and Ghungroo (20 from each) were maintained during hot summer period (May to June) with a mean temperature of about $38^{\circ}C$. The pigs were slaughtered and meat samples from the longissimus dorsi (LD) muscles were analyzed for pH, glycogen and lactate content and mRNA expression. Following 24 h of chilling, LD muscle was also taken from the carcasses to evaluate protein solubility and different meat quality measurements. Results: LWY exhibited significantly (p<0.01) higher plasma cortisol and lactate dehydrogenase concentration than Ghungroo indicating their higher sensitivity to high temperature. LD muscle from LWY pigs revealed lower initial and ultimate pH values and higher drip loss compared to Ghungroo, indicating a faster rate of pH fall. LD muscle of Ghungroo had significantly lower lactate content at 45 min postmortem indicating normal postmortem glycolysis and much slower glycolytic rate at early postmortem. LD muscle of LWY showed rapid postmortem glycolysis, higher drip loss and higher degrees of protein denaturation. Ghungroo exhibited slightly better water holding capacity, lower cooking loss and higher protein solubility. All HSPs (HSP27, HSP70, and HSP90) and MCTs (MCT1, MCT2, and MCT4) in the LD muscle of pigs inclined to increase more in Ghungroo than LWY when exposed to high temperature. Conclusion: Effect of high temperature on the variation of HSPs and MCTs may play a crucial role in thermal tolerance and adaptation to different climatic conditions, pH regulation, muscle acidification, drip loss, protein denaturation and also in postmortem meat quality development.

• 한국습지학회지
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• 제19권2호
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• pp.230-239
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• 2017

최근 한국은 농촌관광의 수요가 증가하고 있지만 유형은 농사체험이 대부분이다. 이러한 상황을 극복하기 위해 생태체험이 도입되고 있다. 농촌 생태체험의 공간인 연못의 콘텐츠, 교육자료 확보가 필요하다. 따라서 본 연구에서는 연못형습지 식물의 전통한방이용형태를 분석하였다. 전통한방은 과거부터 이어져 온 전통지식으로 요즘에도 의학, 약학, 과학 등 다양한 분야에서 활용한다. 분석된 내용은 생태체험 교육자료에 도움이 될 것이다. 조사는 농촌지역 연못형습지 40개소의 식물을 조사하였다. 전통한방이용형태는 특허청에서 운영하는 전통지식포탈의 내용을 제공받아 사용하였다. 40개 연못형습지에서 조사된 식물은 총 108과 457종이며, 그 중 전통한방이용형태에 대한 정보가 있는 종은 53과 314종이다. 이것은 연못형습지 주변의 식물 중 68.8%가 전통한방을 함유하고 있다는 의미다. 이 밖에도 음식, 공예, 전통농업 등의 활용 형태를 포함한다면 70~80% 이상 전통지식을 포함할 것이라 예상한다. 전통한방에 사용 된 314종의 효능은 570가지, 325가지의 병증을 치료하는 효과가 있다. 1종당 평균 4.0(0~20)가지의 효능으로 6.6(0~20)가지의 병증을 치료할 수 있다. 본 연구 자료는 체험, 교육, 의학 등 다양한 분야에 활용되길 기대한다. 그리고 많은 가치가 있는 연못형습지는 계속적으로 보전할 필요가 있다고 판단된다.

• IMMUNE NETWORK
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• 제2권1호
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• pp.41-48
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• 2002

Background: Viral antigens presented on the cell surface in association with MHC class I molecules are recognized by CD8+ T cells. MHC restricted peptides are important in eliciting cellular immune responses. As peptide antigens have a weak immunigenicity, pH-sensitive liposomes were used for peptide delivery to induce effective cytotoxic T lymphocyte (CTL) responses. In the previous study, as the HBx peptides could induce specific CTLs in vitro, we tested whether the HLA-A2/$K^b$ transgenic mice that were immunized by HBx-derived peptides could be protected from a viral challenge. Methods: HBx-peptides encapsulated by pH-sensitive liposomes were prepared. $A2K^b$ transgenic mice were immunized i.m. on days one and seven with the indicated concentrations of liposome-encapsulated peptides. Three weeks later, mice were infected with $1{\times}10^7pfu$/head of recombinant vaccinia virus (rVV)-HBx via i.p. administration. The ovaries were extracted from the mice, and the presence of rVV-HBx in the ovaries was analyzed using human TK-143B cells. IFN-${\gamma}$ secretion by these cells was directly assessed using a peptide-pulsed target cell stimulation assay with either peptide-pulsed antigen presenting cells (APCs), concanavalin A ($2{\mu}g/ml$), or a vehicle. To generate peptide-specific CTLs, splenocytes obtained from the immunized mice were stimulated with $20{\mu}g/ml$ of each peptide and restimulated with peptide-pulsed APC four times. The cytotoxic activity of the CTLs was assessed by standard $^{51}Cr$-release assay and intracellular IFN-${\gamma}$ assay. Results: Immunization of these peptides as a mixture in pH-sensitive liposomes to transgenic mice induced a good protective effect from a viral challenge by inducing the peptide-specific CD8+ T cells. Mice immunized with $50{\mu}g/head$ were much better protected against viral challenge compared to those immunized with $5{\mu}g$/head, whereas the mice immunized with empty liposomes were not protected at all. After in vitro CTL culture by peptide stimulation, however, specific cytotoxicity was much higher in the CTLs from mice immunized with $5{\mu}g/head$ than $50{\mu}g/head$ group. Increase of the number of cells that intracellular IFN-${\gamma}$ secreting cell among CD8+ T cells showed similar result. Conclusion: Mice immunized with XEPs within pH-sensitive liposome were protected against viral challenge. The protective effect depended on the amount of antigen used during immunization. XEP-3-specific CTLs could be induced by peptide stimulation in vitro from splenocytes obtained from immunized mice. The cytotoxic effect of CTLs was measured by $^{51}Cr$-release assay and the percentage of accumulated intracellular IFN-${\gamma}$ secreting cells after in vitro restimulation was measured by flow cytometric analysis. The result of $^{51}Cr$-release cytotoxicity test was well correlated with that of the flow cytometric analysis. Viral protection was effective in immunized group of $50{\mu}g/head$, while in the in vitro restimulation, it showed more spectific response in $5{\mu}g$/head group.

• Clinical and Experimental Reproductive Medicine
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• 제32권4호
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• pp.315-324
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• 2005

Objectives: To compare the efficacy of GnRH antagonist multiple dose protocol (MDP) with that of GnRH agonist long protocol (LP) in controlled ovarian hyperstimulation for in vitro fertilization in patients with high basal FSH (follicle stimulating hormone) level or old age, a retrospective analysis was done. Methods: Two hundred ninety four infertile women (328 cycles) who were older than 41 years of age or had elevated basal FSH level (> 8.5 mIU/mL) were enrolled in this study. The patients had undergone IVF-ET after controlled ovarian hyperstimulation using GnRH antagonist multiple dose protocol (n=108, 118 cycles) or GnRH agonist long protocol (n=186, 210 cycles). The main outcome measurements were cycle cancellation rate, consumption of gonadotropins, the number of follicles recruited and total oocytes retrieved. The number of fertilized oocytes and transferred embryos, the clinical pregnancy rates, and the implantation rates were also reviewed. And enrolled patients were divided into three groups according to their age and basal FSH levels; Group A - those who were older than 41 years of age, Group B - those with elevated basal FSH level (> 8.5 mIU/mL) and Group C - those who were older than 41 years of age and with elevated basal FSH level (> 8.5 mIU/mL). Poor responders were classified as patients who had less than 4 retrieved oocytes, or those with $E_2$ level <500 pg/mL on the day of hCG injection or those who required more than 45 ampules of exogenous gonadotropin for stimulation. Results: The cancellation rate was lower in the GnRH antagonist group than in GnRH agonist group, but not statistically significant (6.8% vs. 9.5%, p=NS). The amount of used gonadotropins was significantly lower in GnRH antagonist group than in agonist group ($34.8{\pm}11.3$ ampules vs. $44.1{\pm}13.4$ ampules, p<0.001). The number of follicles > 14 mm in diameter was significantly higher in agonist group than in antagonist group ($6.7{\pm}4.6$ vs. $5.0{\pm}3.4$, p<0.01). But, there were no significant differences in clinical pregnancy rate (24.5% in antagonist group vs. 27.4% in agonist group, p=NS) and implantation rate (11.4% in antagonist group vs. 12.0% in agonist group, p=NS) between two groups. Mean number of retrieved oocytes was significantly higher in GnRH agonist LP group than in GnRH antagonist MDP group ($5.4{\pm}3.5$ vs. $6.6{\pm}5.0$, p<0.0001). But, the number of mature and fertilized oocytes, and the number of good quality (grade I and II) and transferred embryos were not different between two groups. In each group A, B, and C, the rate of poor response did not differ according to stimulation protocols. Conclusions: In conclusion, for infertile women expected poor ovarian response such as who are old age or has elevated basal FSH level, a protocol including a controlled ovarian hyperstimulation using GnRH antagonist appears at least as effective as that using a GnRH agonist, and may offer the advantage of reducing gonadotropin consumption and treatment period. However, much work remains to be done in optimizing the GnRH antagonist protocols and individualizing these to different cycle characteristics.

• 한국산림과학회지
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• 제94권6호
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• pp.402-409
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• 2005

제주도에 조성한 리기다소나무(Pinus rigida Mill.), 리기테다소나무(Pinus rigida ${\times}$ P. taeda) 채종원내 조성년도가 다른 6개 식재지에서 푸사리움가지마름병(pitch canker) 피해를 입은 지 7년째 되는 잔존 임목의 피해를 개체별로 판정하여 채종원 및 수종간 피해도 차이를 비교하였으며 이들 중 표현형에 의해 선발한 내병성, 이병성 선발목의 반형매 차대 2년생 묘목을 리기다소나무와 해송(P. thunbergii Parl.)에서 분리한 푸사리움가지마름병균(Fusarium circinatum)으로 인공접종 한 뒤 기주의 감염성 여부를 검정하였고 균주들의 병원성 여부를 확인하였다. 조사한 특성 중 두 채종원(상효, 한남)내 리기테다소나무 4 식재지간에 SC(줄기궤양)에서 통계적 유의성(${\chi}^2=7.76$; P=0.05)이 있는 것으로 나타났다. 상효채종원내에서 리기다소나무가 리기테다소나무에 비해 수관피해도(top kill)와 가지끝 마름증상(branch tip symptoms)에서 피해도가 높은 것으로 나타났다. 인공접종 결과 내병성 후보목 차대가 이병성 후보목 차대에 비해 14% 높은 고사율을 보였는데, 이는 화분수의 영향이나 표현형에 의한 후보목 선발에서 온 결과로 사료된다. 처리균주 5개 중 C-6-L(9)과 C-6-L(19) 균주가 각각 68%, 60%의 고사율을 보여 차후 추진할 대규모 인공접종용 균주로 사용할 수 있을 것이다. 인공접종 실험을 통해 1차 선발된 내병성 후보목 개체들은 푸사리움가지마름병 저항성 품종 육종을 위한 유전적, 생화학적 기초 연구에 유용한 재료로 이용 될 수 있을 것으로 본다.