• 한국식품영양과학회지
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• 제37권11호
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• pp.1507-1514
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• 2008

본 연구에서는 산업적으로 사용될 수 있는 안정하고 활성이 높은 cholesterol oxidase를 생산하는 균주를 얻기 위해 토양 미생물로부터 균주를 선별하였다. 선별된 균주에 대하여 세포외 효소 활성을 측정한 결과 BEN 115로 명명한 균주가 가장 높은 효소 활성도를 나타내었다. 이 균주는 형태학적, 생리학적 특성과 배양형태 및 G＋C 함량을 분석한 결과 Nocardia속으로 확인되었다. 최적 효소 생산 조건을 조사한 결과 기존 yeast malt extract broth 조성인 0.4% yeast extract, 0.4% glucose, 1% malt extract에서 가장 높은 활성을 나타냈다. 본 균주에서 생산된 extracellular cholesterol oxidase는 SDS-PAGE와 Western blot 결과 분자량이 55, 57 kDa인 두 종류의 효소가 존재하는 것으로 나타났다. BEN 115에 대한 효소학적 특성을 연구한 결과 온도 안정성은 $55^{\circ}C$까지 효소 활성이 유지되었고, pH 안정성은 pH $3.5{\sim}9.5$의 범위까지 안정한 것으로 나타났으며 최적 온도와 최적 pH는 각각 35oC와 pH 5.5인 것으로 나타났다. 또한 detergent(Triton X-100, Triton X-114 그리고 Tween 80) 첨가 시 효소 활성이 첨가하지 않은 대조군보다 약 $1.6{\sim}2.0$배 증가하는 것으로 나타났다. Campesterol, sitosterol 그리고 stigmasterol에 대한 기질 특이성은 cholesterol(100%)과 상대적으로 비교 시 각각 50%, 50% 그리고 27%의 기질 특이성을 나타내었다.

• 한국수산과학회지
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• 제39권2호
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• pp.100-105
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• 2006

This study was conducted to evaluate the effects of extruded pellet (EP) diets, as compared to a raw fish moist pellet (MP) diet for olive flounder, Paralichthys olivaceus, grown in commercial-scale aquaculture for 1 year. Four diets with duplication per diet were formulated for this experiment: two experimental EP diets (EP1 and EP2), one commercial EP diet (CEP), and a raw fish MP diet (MP). The MP diet consisted of 80% frozen horse mackerel and 20% commercial binder meal. Fish weighing $30.1{\pm}0.1 g$ ($mean{\pm}SD$) were distributed randomly to each aquarium as a group of 2,600 fish. Weight gain (WG) and feed efficiency ratio (FER) of fish fed EP2 and MP were higher (P<0.05) than those of fish fed CEP, while those of fish fed EP1 did not differ (P>0.05) from those fed EP2 and MP. However, fish fed the MP diet had a higher survival rate than fish fed the other diets. Fish fed EP2 had higher serum, phospholipids and total protein levels, and lower levels of serum glutamic-oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), and total cholesterol than fish fed MP (P<0.05). Dorsal muscle and liver proteins and lipid of fish fed EP1 were higher (P<0.05) than those of fish fed CEP, while those of fish fed EP1 and MP did not differ from those of fish fed EP2 (P>0.05). These results strongly suggest that EP1 could be developed to replace MP for grow-out stage production of olive flounder without adverse effects on growth performance.

• The Korean Journal of Physiology and Pharmacology
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• 제20권1호
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• pp.101-109
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• 2016

Reducing $[Mg^{2+}]_o$ to 0.1 mM can evoke repetitive $[Ca^{2+}]_i$ spikes and seizure activity, which induces neuronal cell death in a process called excitotoxicity. We examined the issue of whether cultured rat hippocampal neurons preconditioned by a brief exposure to 0.1 mM $[Mg^{2+}]_o$ are rendered resistant to excitotoxicity induced by a subsequent prolonged exposure and whether $Ca^{2+}$ spikes are involved in this process. Preconditioning by an exposure to 0.1 mM $[Mg^{2+}]_o$ for 5 min inhibited significantly subsequent 24 h exposure-induced cell death 24 h later (tolerance). Such tolerance was prevented by both the NMDA receptor antagonist D-AP5 and the L-type $Ca^{2+}$ channel antagonist nimodipine, which blocked 0.1 mM $[Mg^{2+}]_o$-induced $[Ca^{2+}]_i$ spikes. The AMPA receptor antagonist NBQX significantly inhibited both the tolerance and the $[Ca^{2+}]_i$ spikes. The intracellular $Ca^{2+}$ chelator BAPTA-AM significantly prevented the tolerance. The nonspecific PKC inhibitor staurosporin inhibited the tolerance without affecting the $[Ca^{2+}]_i$ spikes. While $G{\ddot{o}}6976$, a specific inhibitor of $PKC{\alpha}$ had no effect on the tolerance, both the $PKC{\varepsilon}$ translocation inhibitor and the $PKC{\zeta}$ pseudosubstrate inhibitor significantly inhibited the tolerance without affecting the $[Ca^{2+}]_i$ spikes. Furthermore, JAK-2 inhibitor AG490, MAPK kinase inhibitor PD98059, and CaMKII inhibitor KN-62 inhibited the tolerance, but PI-3 kinase inhibitor LY294,002 did not. The protein synthesis inhibitor cycloheximide significantly inhibited the tolerance. Collectively, these results suggest that low $[Mg^{2+}]_o$ preconditioning induced excitotoxic tolerance was directly or indirectly mediated through the $[Ca^{2+}]_i$ spike-induced activation of $PKC{\varepsilon}$ and $PKC{\xi}$, JAK-2, MAPK kinase, CaMKII and the de novo synthesis of proteins.

• 대한한방내과학회지
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• 제29권1호
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• pp.130-148
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• 2008

Objective : This study was designed to investigate the effect of the water extract of Gamisamgibopae-tang(GMSGBPT), an oriental herbal formulation, on the growth of NCI-H460 and A549 human non-small-cell lung cancer cell lines. Methods : Cytotoxicity and cell morphology were evaluated by MTT assay and inverted microscope, respectively. Apoptosis was detected using agarose gel electrophoresis and flow cytometer. The expression levels of mRNAs and proteins of target genes were determined by RT-PCR and western blot analyses, respectively Result and Conclusion : We found that exposure of A549 cells to GMSGBPT resulted in the growth inhibition in a dose-dependent manner as measured by MTT assay, but GMSGBPTdid not affect the growth of NCI-H460 cells. The anti-proliferative effect of GMSGBPT treatment in A549 cells was associated with morphological changes, formation of apoptotic bodies and DNA fragmentation, and flow cytometry analysis confirmed that GMSGBPT treatment increased the populations of apoptotic-sub G1 phase. Growth inhibition and apoptotic cell death by GMSGBPT were connected with a up-regulation of cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) mRNA and protein in a tumor suppressor p53-independent fashion. However GMSGBPT treatment did not affect other growth regulation-related genes such as early growth response-1 (Egr-1), nonsteroidal anti-inflammatory drug (NSAID)-activated gene-1 (NAG-1), inducible nitric oxide synthase (iNOS), cyclooxygenases (COXs), telomere-regulatory factors in A549 orNCI-H460 cells. Taken together, these findings partially provide novel insights into the possible molecular mechanism of the anti-cancer activity of GMSGBPT.

• 한국식품과학회지
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• 제17권4호
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• pp.248-252
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• 1985

비이용 자원의 효율적인 이용의 가능성을 검토하기 위하여 앵두씨의 지방질 조성 및 단백질 패턴을 분석한 결과는 다음과 같다. 일반성분 중 조지방질과 조단백질의 함량은 각각 40.38% 및 26.59%였다 앵두씨의 지방질은 중성지방질이 95.4%인 반면에 복합지방질은 4.51%에 불과하였으며, 중성지방질의 성분으로는 트리-글리세리드가 89.86%로서 대부분이었고. 스테롤 4.14%. 모노-글리세리드 2.98%, 스테롤 에스테르 1.77%. 유리지방산 1.07% 및 디 -글리세리드 0.18% 였다. 지방산 조성은 총 지방질, 중성지방절 및 트리 글리세리드에서 올레산(65.06$\sim$66.05%)와 리놀레산(26.56$\sim$28.40%)이 주된 구성 지방산이었고, 당지방질파 인지방질의 경우에는 올FP산(40.55$\sim$51.46%)의 함량이 다른 획분의 함량에 비해 적은 반면, 팔미트산(17.64$\sim$21.43%)과 스테아르산(4.14$\sim$8.07%) 의 함량이 높았으며, 소량의 라오르산도 함유되어 있었다. 염용해성 단백질의 추출율은 약 60%였으며, 분획된 주단백질의 수득율은 약 46.5% 였고 전기영동 결과 7개의 밴드가 확인되었다.

• 한국식품저장유통학회지
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• 제9권1호
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• pp.67-73
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• 2002

수입 참깨가루로 제조한 참기름의 품질과 위생적 안정성에 대한 기초자료를 얻고자 통참깨와 참깨가루를 원료로한 참기름의 일반성분, 화학적 특성, 지방산, sesamol, sesamolin, sesamin 함량 및 AOM에 의한 유도기간 등을 비교 조사하였다. 일반성분 중 수분은 통참깨에 비해 참깨가루가 월등히 낮게 나타났으며, 단백질은 거의 차이가 없었다. 참기름의 화학적 특성으로 산가와 검화가는 참깨가루가 통참깨에 비해 높게 나타났으며, 요오드가는 통참깨군보다 낮았다. 참기름의 지방산 조성에서 linolenic acid과 linoleic acid의 함량은 각각 222.44~144.14, 2713~1776.46 $\mu\textrm{g}$/mL이었고, linoleic acid와 ${\gamma}$-linoleic acid의 함량이 통참깨 참기름에 비해 참깨가루 참기름의 함량이 낮았다. Lignan 물질 중 sesamol 함량은 통참깨 참기름이 참깨가루 참기름보다 함량이 높았고, 인도산 통참깨 참기름의 sesamol 함량이 가장 높았다. Sesamin, sesamolin 함량은 한국만 통참깨 참기름이 가장 높았다. AOM에 의한 산화유도시간은 북한산, 중국산 참깨가루가 각각 6.67, 13.35 시간으로 나타나 통참깨군보다 산화안정성이 낮은 것으로 나타났다.

• 대한한의학방제학회지
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• 제23권2호
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• pp.189-198
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• 2015

Objectives : Sparganii Rhizoma is frequently used in traditional herbal medicine for treatment of blood stasis, amenorrhea and functional dyspepsia and has been reported to exhibit anti-oxidant, anti-proliferation and anti-angiogenesis peoperties. In this study, we investigated the cytoprotective effect and underlying mechanism of Sparganii Rhizoma water extract (SRE) against oxidative stress-induced mitochondrial dysfunction and apoptosis in hepatocyte. Methods : To determine the effects of SRE on oxidative stress, we induced synergistic cytotoxicity by co-treatment of arachidonic acid (AA) and iron in the HepG2 cell, a human derived hepatocyte cell line. Results : Treatment of SRE increased relative cell viability and altered the expression levels of apoptosis-related proteins such as Bcl-xL, Bcl-2 and procaspase-3. And SRE also inhibited the mitochondrial dysfunction and excessive reactive oxygen species production induced by AA+iron. In addition, SRE activated of AMP-activated protein kinase (AMPK), a potential target for cytoprotection, by increasing the phosphorylation of AMPKα at Thr-172. Morever, SRE increased phosphorylation of acetyl-CoA carboxylase, a direct downstream target of AMPK. Conclusion : These results indicated that SRE has the ability to protect against oxidative stress-induced hepatocyte damage, which may be mediated with AMPK pathway.

• 한국식품영양과학회지
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• 제37권1호
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• pp.117-123
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• 2008

국내에서 생산된 감자 중 세풍, 남서, 수미, 조풍 및 대서의 5가지 품종에 대하여 단백질 profile 및 아미노산 조성을 분석하였다. 총 질소함량은 $1.27{\sim}1.64%$이었으며, 남서가 높았고 수미가 낮게 나타났다. 아미노산 조성은 품종 간에 유의적인 차이가 있었다. 한편, 주요 감자 단백질은 papatin(40 kDa), trypsin inhibitor(20 kDa) 및 protease inhibitor(15 kDa)이었으며, 이들의 함량은 각각 $22.16{\sim}25.81%$, $25.22{\sim}20.91%$ 및 $14.12{\sim}25.23%$이었다. Papatin 함량은 조풍, 세풍, 수미감자가 높은 함량을 보인 반면, trypsin inhibitor는 조풍감자가 5.22%로 가장 낮은 함량을 보였다. Protease inhibitors인 20 kDa와 15 kDa를 합한 값은 $24.7{\sim}35.0%$이었으며, 세풍이 가장 적었고 조풍에 가장 많이 함유되어 있었다.

• International Journal of Oral Biology
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• 제31권4호
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• pp.141-148
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• 2006

The effects of adenosine triphosphate(ATP) on salivary glands have been recognized since 1982. The presence of purinergic recepetors(P2Rs) that mediate the effects of ATP in various tissues, including parotid and submandibular salivary gland, has been supported by the cloning of receptor cDNAs and the expression of the receptor proteins. P2Rs have many subtypes, and the activation of these receptor subtypes increase intracellular $Ca^{2+}$, a key ion in the regulation of the secretion in the salivary gland. The apical pores of taste buds in circumvallate and foliate papillae are surrounded by the saliva from von Ebner salivary gland(vEG). Thus, it is important how the secretion of vEG is controlled. This study was designed to elucidate the roles of P2Rs on salivary secretion of vEG. Male Sprague-Dawley rats (about 200 g) were used for this experiment. vEG-rich tissues were obtained from dissecting $500-1,000\;{\mu}m$ thick posterior tongue slices under stereomicroscope view. P2Rs mRNA in vEG acinar cells were identified with RT-PCR. To observe the change in intracellular $Ca^{2+}$ activity, we employed $Ca^{2+}-ion$ specific fluorescence analysis with fura-2. Single acinar cells and cell clusters were isolated by a sequential trypsin/collagenase treatment and were loaded with $10\;{\mu}M$ fura -2 AM for 60 minutes at room temperature. Several agonists and antagonists were used to test a receptor specificity. RT-PCR revealed that the mRNAs of $P2X_4$, $P2Y_1$, $P2Y_2$ and $P2Y_3$ are expressed in vEG acinar cells. The intracellular calcium activity was increased in response to $10\;{\mu}M$ ATP, a P2Rs agonist, and 2-MeSATP, a $P2Y_1$ and $P2Y_2R$ agonist. However, $300\;{\mu}M\;{\alpha}{\beta}-MeATP$, a $P2X_1$ and $P2X_3R$ agonist, did not elicit the response. The responses elicited by $10\;{\mu}M$ ATP and UTP, a $P2Y_2R$ agonists, were maintained when extracellular calcium was removed. $10\;{\mu}M$ suramin, a P2XR antagonist, and reactive blue 2, a P2YR antagonist, partially blocked ATP-induced response. However, when extracellular calciums were removed, suramin did not abolish the responses elicited by ATP. These results suggest that P2Rs play an important role in salivary secretion of vEG acinar cells and the effects of ATP on vEG salivary secretion may be mediated by $P2X_4$, $P2Y_1$, $P2Y_2$, and/or $P2Y_3$.

• 대한본초학회지
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• 제30권6호
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• pp.7-15
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• 2015

Objectives : Rosa laevigata Michx. has been used for the treatment of renal disease in traditional Korean medicine. In this study, we investigated cytoprotective effect of R. laevigata water extract (RLE) against oxidative stress induced by arachidonic acid (AA) + iron.Methods : To evaluate the protective effects of RLE against AA + iron-induced oxidative stress in HepG2 cell, cell viability and changes on apoptosis-related proteins were assessed by MTT and immunoblot analyses. The effects of RLE on reduced glutathione level, production of reactive oxygen species and mitochondrial membrane potential were also monitored. Furthermore, to verify underlying molecular mechanism, NF-E2-related factor 2 (Nrf2) was examined by immunoblot analysis. Additionally, Nrf2 transactivation and its downstream target genes expression were also determined by reporter gene and realtime RT-PCR analyses.Results : RLE pretreatment (30-300 μg/ml) prevented cells from AA + iron-mediated cell death in a concentration dependent manner. In addition, 100 μg/ml RLE inhibited AA + iron-induced glutathione depletion, reactive oxygen species production and mitochondrial dysfunction. RLE accumulated nuclear Nrf2 and also transactivated Nrf2, which was evidenced by antioxidant response element- and glutathione S-transferase A2-driven luciferase activities and mRNA level of glutamate-cysteine ligase catalytic subunit, NAD(P)H:quinone oxidoreductase 1 and sestrin 2. Moreover, protective effect of RLE against AA + iron was abolished in Nrf2 knockout cells.Conclusions : These results indicate that RLE has the ability to protect hepatocyte against oxidative stress through Nrf2 activation.