• KSBB Journal
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• 제30권6호
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• pp.291-295
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• 2015

The limited productivity of natural shell matrix proteins has hampered the investigation of their biochemical properties and practical applications, although biominerals in nature obtained by organic-inorganic assemblies have attractive mechanical and biological properties. Here, we prepared a vector for the expression of a fusion protein of a shell matrix protein from Pinctada fucata (named as GRP_BA) with the GRGDSP residue. The fusion protein of BA-RGD was simply produced in E. coli and purified through sequential steps including the treatment with $CaCl_2$ and EDTA solution for cell membrane washing, mechanical cell disruption and the application of non-ionic surfactant of Triton X-100 for BA-RGD inclusion body washing. The production yield was approximately 60 mg/L, any other protein band was not observed in SDS-PAGE and it was estimated that above 97% endotoxin was removed compared to the endotoxin level of whole cell. This study showed this simple and easy purification approach could be applied to the purification of BA-RGD fusion protein. It is expected that the protein could be utilized for the preparation of biominerals in practical aspects.

• 대한골관절종양학회지
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• 제10권1호
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• pp.29-33
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• 2004

원위 요골의 거대 세포종은 빈도가 많지는 않다. 통상적으로 골 소파술 및 골 시멘트 충전술로 치료하지만, 재발한 경우나 처음부터 골피질 파괴가 심하고 관절 침범이 있을 경우에는 일괄 절제(en bloc resection) 후 근위 비골을 이용하여 재건하는 술 식이 많이 이용되어 왔다. 본 연구는 고식적 술 식으로 치료한 후 국소 재발한 원위 요골의 거대 세포종 환자에서, 근위 비골을 이용한 재건술을 시행하여도 일차 술 식 시 오염의 범위가 심하여 다시 재발할 가능성이 높아 초고분자량 폴리에틸렌(ultrahigh molecular weight polyethylene, UHMWPE)과 골수강내 고정물 및 골 시멘트를 조합하여 원위 요골을 재건한 1례를 보고 하고자 한다.

• Toxicological Research
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• 제31권4호
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• pp.331-337
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• 2015

Synthetic pyrethroids (SPs) are the most common pesticides which are recently used for indoor pest control. The widespread use of SPs has resulted in the increased exposure to wild animals and humans. Recently, some SPs are suspected as endocrine disrupting chemicals (EDCs) and have been assessed for their potential estrogenicity by adopting various analyzing assays. In this study, we examined the estrogenic effects of lambda-cyhalothrin (LC) and cypermethrin (CP), the most commonly used pesticides in Korea, using BG-1 ovarian cancer cells expressing estrogen receptors (ERs). To evaluate the estrogenic activities of two SPs, LC and CP, we employed MTT assay and reverse-transcription polymerase chain reaction (RT-PCR) in LC or CP treated BG-1 ovarian cancer cells. In MTT assay, LC ($10^{-6}M$) and CP ($10^{-5}M$) significantly induced the growth of BG-1 cancer cells. LC or CP-induced cell growth was antagonized by addition of ICI 182,720 ($10^{-8}M$), an ER antagonist, suggesting that this effect appears to be mediated by an ER-dependent manner. Moreover, RT-PCR results showed that transcriptional level of cyclin D1, a cell cycle-regulating gene, was significantly up-regulated by LC and CP, while these effects were reversed by co-treatment of ICI 182,780. However, p21, a cyclin D-ckd-4 inhibitor gene, was not altered by LC or CP. Moreover, $ER{\alpha}$ expression was not significantly changed by LC and CP, while down-regulated by E2. Finally, in xenografted mouse model transplanted with human BG-1 ovarian cancer cells, E2 significantly increased the tumor volume compare to a negative control, but LC did not. Taken together, these results suggest that LC and CP may possess estrogenic potentials by stimulating the growth of BG-1 ovarian cancer cells via partially ER signaling pathway associated with cell cycle as did E2, but this estrogenic effect was not found in in vivo mouse model.

• Biomolecules & Therapeutics
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• 제12권1호
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• pp.25-33
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• 2004

In this study we investigated whether ATP loss was involved in the potentiated death of immunostimulated rat primary astrocytes in glucose-deprived condition. Rat primary astrocytes immunostimulated with LPS plus IFN-${\gamma}$ for 48 h underwent death upon glucose deprivation, which dependent on the production of peroxynitrite. Intracellular ATP level synergistically decreased by glucose deprivation in immunostimulated astrocytes but not in control cells, and the loss of ATP occurred well ahead of the LDH release. The synergistic cell death and ATP loss by immunostimulation and glucose deprivation were inhibited by iNOS inhibitor (L-NAME and L-NNA) or peroxynitrite decomposition catalyst (also a superoxide anion scavenger), Mn(III)tetrakis(N-methyl-4'-pyridyl)porphyrin (MnTMPyP). Exogenous addition of peroxynitrite generator, SIN-l timedependently induced ATP loss and cell death in the glucose-deprived astrocytes. Depletion of intracellular glutathione (GSH) and dis겨ption of mitochondrial transmembrane potential (MTP) were also observed under same conditions. Supply cellular ATP by the addition of exogenous adenosine or ATP during glucose deprivation inhibited ATP depletion, GSH depletion, MTP disruption and cell death in SIN-l treated or immunostimulated astrocytes. This study showed that perturbation in the regulation of intracellular ATP level in immunostimulated astrocytes might make them more vulnerable to energy challenging stimuli.

• Parasites, Hosts and Diseases
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• 제49권1호
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• pp.1-8
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• 2011

The pathogenesis and pathophysiology of Acanthamoeba infections remain incompletely understood. Phospholipases are known to cleave phospholipids, suggesting their possible involvement in the host cell plasma membrane disruption leading to host cell penetration and lysis. The aims of the present study were to determine phospholipase activities in Acanthamoeba and to determine their roles in the pathogenesis of Acanthamoeba. Using an encephalitis isolate (T1 genotype), a keratitis isolate (T4 genotype), and an environmental isolate (T7 genotype), we demonstrated that Acanthamoeba exhibited phospholipase $A_2$ (PLA$_2$). and phospholipase D (PLD) activities in a spectrophotometry-based assay. Interestingly, the encephalitis isolates of Acanthamoeba exhibited higher phospholipase activities as compared with the keratitis isolates, but the environmental isolates exhibited the highest phospholipase activities. Moreover, Acanthamoeba isolates exhibited higher PLD activities compared with the PLA$_2$. Acanthamoeba exhibited optimal phospholipase activities at $37^{\circ}C$ and at neutral pH indicating their physiological relevance. The functional role of phospholipases was determined by in vitro assays using human brain microvascular endothelial cells (HBMEC), which constitute the blood-brain barrier. We observed that a PLD-specific inhibitor, i.e., compound 48/80, partially inhibited Acanthamoeba encephalitis isolate cytotoxicity of the host cells, while PLA$_2$-specific inhibitor, i.e., cytidine 5'-diphosphocholine, had no effect on parasite-mediated HBMEC cytotoxicity. Overall, the T7 exhibited higher phospholipase activities as compared to the T4. In contract, the T7 exhibited minimal binding to, or cytotoxicity of, HBMEC.

• Journal of Microbiology
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• 제39권4호
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• pp.286-292
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• 2001

During mitosis. the proper segregation of duplicated chromosomes is corrdinated by a spindle check-point. The bifurcated spindle checkpoint blocks cell cycle progression at metaphase by monitoring unattached kinetochores and inhibits mitotic exit in response to the misorientation of the mitotic spin- dle Ibd1p/Bfa1p is a spindle checkpoint regulator of budding yeast in the Bub2p checkpoint pathway for mitotic exit and its disruption abolishes mitotic arrest when proper organization of the mitotic spin-dls inhibited. Ibd1p/Bfa1p localizes to the spindle pole body, a microtublue-organizing center in yeast, and its overexpression arrests the cell cycle in 80% of cells with an enlarged budy at mitosis and in 20 % of cells with multiple buds. In this study, we found that the C-terminus of Ibd1p/Bfa1p phys-ically interacts with Skp1p, a key component of SCF (Skp1/cullin/F-box) complex for ubiquition-medi-ated proteolysis of cel cycle regulatores as well as an evolutionally conserved kinetochore protein for cell cycle progression. A putative F-box motif was found in the C-terminus of Ibd1p/Bfa1p and its function was investigated by making mutants of conserved residues in the motif. These Ibd1p/Bfa1p mutants of a putative F-box interacted with SKp1p in vitro by two-hybrid assays as wild type Ibd1p/Bfa1p. Also these Ibd1p/Bfa1p utants displayed the overexpression phenotypes of wild type Ibd1p, when over-expressed under inducible promoters . These results suggest that a putative F-box motif of Ibd1p/Bfa1p is not essential for the interaction with SKp1p and its function in mitotic exit and cytokinesis.

• 한국패류학회지
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• 제29권1호
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• pp.7-13
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• 2013

본 연구는 납에 노출된 바지락, Ruditapes philippinarum 의 생물학적 반응을 알아보고자 하였다. 실험기간은 4주였으며, 실험구는 대조구 1개와 납 노출구 3개 (0.25, 0.50 and 1.00 mg/l) 였다. 실험 결과 납은 바지락의 생존율과 산소 소비율의 저하 및 기관계의 조직학적 변성을 유도하는 것이 확인 되었다. 납 노출구에서 산소소비율은 25-72% 감소하였다. 기관계의 조직학적 분석 결과, 외투막 상피층과 결합조직층의 변성, 아가미 점액세포의 증가와 상피세포의 괴사, 발에서는 상피층의 붕괴, 점액세포의 감소 및 혈림프동의 확장과 결합조직층의 변성을 나타냈다. 소화맹낭의 소화선세관에서는 호염기성세포와 상피세포의 위축 및 농도 의존적으로 lipofuscin의 축적이 확인되었다.

• Molecules and Cells
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• 제41권8호
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• pp.733-741
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• 2018

Mutations in spliceosome components have been implicated in carcinogenesis of various types of cancer. One of the most frequently found is U2AF1 S34F missense mutation. Functional analyses of this mutation have been largely limited to hematological malignancies although the mutation is also frequently seen in other cancer types including lung adenocarcinoma (LUAD). We examined the impact of knockdown (KD) of wild type (wt) U2AF1 and ectopic expression of two splice variant S34F mutant proteins in terms of alternative splicing (AS) pattern and cell cycle progression in A549 lung cancer cells. We demonstrate that induction of distinct AS events and disruption of mitosis at distinct sub-stages result from KD and ectopic expression of the mutant proteins. Importantly, when compared with the splicing pattern seen in LUAD patients with U2AF1 S34F mutation, ectopic expression of S34F mutants but not KD was shown to result in common AS events in several genes involved in cell cycle progression. Our study thus points to an active role of U2AF1 S34F mutant protein in inducing cell cycle dysregulation and mitotic stress. In addition, alternatively spliced genes which we describe here may represent novel potential markers of lung cancer development.

• Advances in environmental research
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• 제3권4호
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• pp.307-319
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• 2014

Four chickpea cultivars viz. kabuli (Pusa 1088 and Pusa 1053) and desi (Pusa 1103 and Pusa 547) differing in sensitivity to high temperature conditions were analyzed in earthern pot (30 cm) at different stages of growth and development in the year of 2010 and 2011. Pusa-1053 (kabuli type) showed maximum photosynthetic rate and least by Pusa-547 (desi type), whereas maximum cell membrane thermostability were recorded in Pusa-1103 and minimum in Pusa-1088. Among the treatments, the plants grown under elevated temperature conditions had produced 13.01% more significant data in comparison to plants grown under continuous natural conditions. Stomatal conductance were reduced 44.25% under elevated temperature conditions than natural conditions, whereas 35.56%, when plants grown under initially natural conditions upto 30DAS, then 30-60DAS elevated temperature and finally shifted to natural conditions till harvest. In case of Pusa-1103, stomatal conductance was maximum as compared to rest of 2.7% from Pusa-1053, 8.9% from Pusa-1088, and 10.3% in Pusa-547 throughout the study. Plants grown under continuous elevated temperature conditions had produced 15.30% and 15.32% more significant membrane thermostability index in comparison to continuous natural conditions at vegetative stage and 19.40% and 18.44% at flowering stage, while the better response was recorded at pod formation stage. Pusa-1053 had given 2.8% more membrane thermostability index than Pusa-1088 and Pusa-1103 had given 1.6% more membrane thermostability index than Pusa-547 in the present study. The membrane disruption caused by high temperature may alter water ion and inorganic solutes movement, photosynthesis and respiration. Thus, thermostability of the cell membrane depends on the degree of the electrolyte leakage.

• 한국통신학회논문지
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• 제36권6B호
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• pp.697-709
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• 2011

차량과 같은 이동환경에서 무선 인터넷 서비스를 사용할 때, 불필요하게 발생하는 핸드오버는 서비스 품질 저하와 시그널링 오버헤드를 유발하여 실시간 멀티미디어 서비스 제공에 단점으로 작용한다. 본 논문에서는 Mobile Node (MN)가 일정한 이동패턴을 가지고 있을 때, Mobility behavior profile을 이용하여 생성한 DTMC을 이용하여 핸드오버의 발생 횟수를 감소시켜 무선 인터넷 서비스를 향상시킬 수 있는 핸드오버 방법을 제안한다. 기존 핸드오버 방법을 반복적으로 수행하며 Mobility behavior profile을 학습하고, Mobility behavior profile 에 충분한 양이 학습되면 Mobility behavior profile을 이용하여 DTMC의 1-step & 2-step transition probability matrix를 생성한다. 그 이후에는 DTMC의 1-step & 2-step transition probability matrix를 이용하여 핸드오버를 수행하며 Mobility behavior profile을 계속 업데이트한다. 4개의 Mobility model에서의 실험을 통하여 평균 핸드오버 횟수와 평균 RSSI값, 그에 따른 Throughput을 비교한다.