Beauvericin (BEA) is a cyclohexadepsipeptide produced by the fungus Beauveria bassiana. And it has been reported to have very effective anti-cancer activity. However, the mechanistic studies of BEA on cytotoxicity on cancer cells have not been fully understood. In this study, we examined the cytotoxicity of BEA on the C6 (rat glioma cell line). The cell viabilities of C6 cells, MDA-MB 231 (breast cancer cell line), and HeLa (cervical cancer cell line) were decreased by treatment of BEA. In addition, BEA-treated cells showed membrane blebbings and buddings, which indicates that BEA decreased cell viability by inducing apoptosis. Additionally, we found that the level of apoptosis inducing molecules such as caspase 3, caspase 9 of BEA treated cells was increased and apoptosis regulating molecules (Bcl-2) was decreased. We also found that the activated STAT3 and Src was decreased in BEA-treated condition. In further study, we are going to find out how the BEA can influence Src kinase activity.

4종 해충(복숭아혹진딧물, 점박이응애, 아메리카잎굴파리, 대만총채벌레)에 이온화에너지인 전자빔과 X-ray를 각각 조사하여 이들 해충 의 발육과 생식에 미치는 영향을 비교하여 억제선량을 결정하였다. 복숭아혹진딧물 약충에 전자빔과 X-ray 조사 시, 전자빔의 경우에는 100 Gy, X-ray의 경우에는 30 Gy에서 우화성충의 산자가 완전히 억제되었다. 복숭아혹진딧물 성충의 경우 200 Gy (전자빔)와 50 Gy (X-ray)에서 각각 F1 세대 약충의 우화가 억제되었다. 그러나 이들 두 이온화에너지 모두 성충수명에는 영향을 주지 않았다. 점박이응애의 알에서는 전자빔 150 Gy와 X-ray 50 Gy 선량에서 알의 부화가 완전히 억제되었다. 아메리카잎굴파리 번데기에서는 X-ray 조사선량이 증가할수록 우화율이 감소하 였다. 대만총채벌레 성충에 조사 시 전자빔의 250 Gy와 X-ray의 200 Gy 선량에서 F1 세대 알의 부화가 완전히 억제되었다.

In this study, to further understand the mechanism of animal growth and to develop a miniature transgenic animal model, we constructed and tested tetracycline-inducible RNAi system using shRNA targeting the mRNA of mouse insulin-like growth factor (mIGF-1) or mouse growth hormone receptor (mGHR) gene. Quantitative real-time PCR analysis of mouse liver cell (Hepa1c1c7) cells transfected with these vectors showed 85% or 90% of expression inhi-bition effect of IGF-1 or GHR, respectively. In ELISA analysis, the protein level of IGF-1 in the cells expressing the shRNA targeting IGF-1 mRNA was reduced to 26% of non-transformed control cells. Unexpectedly, in case of using shRNA targeting GHR, the IGF-1 protein level was decreased to 75% of control cells. Further experiments are needed to explain the lower interference effect of GHR shRNA in IGF-1 protein. Accumulated knowledge of this approach could be applicable to a variety of related biological area including gene function study, gene therapy, development of miniature animals, etc.

Cr3+ 및 Se을 첨가하여 수경 재배한 치커리 3종의 에탄 올 추출물을 각각 제조하여 추출물별 항산화 활성을 비교 검토하였다. 그 결과 양액만으로 재배한 치커리보다 양액에 Se이나 Cr3+을 첨가한 치커리 추출물에서 총 폴리페놀 함량, 총 플라보노이드 함량 및 FRAP 환원능이 증가하였으며, DPPH 라디칼 소거능 및 ABTS 라디칼 소거능도 증가하였다. 특히 Se을 첨가한 치커리보다 Cr3+을 첨가한 치커리에서 더 우수한 항산화 활성을 나타내었다. 또한 Cr3+ 을 첨가한 CLER 및 CLE의 경우 α-glucosidase 저해활성이 있음을 알 수 있었다. 따라서 치커리 추출물의 수경재배 과정에서 양액에 Cr3+, Se의 첨가는 치커리의 항산화 활성 및 α-glucosidase 저해활성 증진에 영향을 주는 것으로 확인되었다.

곤충병원성세균 Xenorhabdus nematophila (Xn)의 일부 대사물질은 대상 곤충의 phospholipase A2 (PLA2)를 억제하여 아이코사노이드 생합성 활성을 저해시킨다. 그러나 이들 세균 대사물질이 억제하는 곤충의 PLA2에 대해서는 알려져 있지 않다. Xn의 배양액에서 화학구조가 동정된 8 가지 대사물질들은 두 종의 나비목 배추좀나방(Plutella xylostella)과 파밤나방(Spodoptera exigua)의 유충에 대하여 살충 활성을 보유했다.특별히 이들 물질은 모두 Bacillus thuringiensis (비티)의 살충력을 크게 향상시켰다. 파밤나방의 세포성 인지질 분해효소(SecPLA2)를 클로닝하고 대장균에서 과발현시켰다. 분리된 SecPLA2를 지방체에서 얻은 인지질과 반응시켰을 때 여러 다가불포화지방산을 해리시켰다. 이 효소활성이 Xn 유래 대사물질들에 의해 뚜렷이 억제되었다. 또한 SecPLA2에 대한 억제효과와 비티 살충력 상승효과 사이에 정의 상관관계를 보였다. 본 연구는 SecPLA2가 Xn 대사물질의 억제 대상 분자 종말점 가운데 하나라고 제시하고 있다.

Chios Gum Mastic (CGM) is a natural resin extracted from the leaves of Pistacia lentiscus, a plant endemic to the Greek island of Chios. It has been used by traditional healers, and it has antibacterial, antifungal properties, and therapeutic benefits for the skin. The CGM reduces the formation of dental plaque and bacterial growth in oral saliva, and recent studies have demonstrated the role of antioxidant activity of CGM. Although CGM has been widely investigated, its protective effect against oxidative-damage to keratinocytes, as well as the relationship between CGM and autophagy, has not been investigated. The aim of this study was to assess the protective effect of CGM against H2O2-induced oxidative stress and to evaluate the autophagic features induced by CGM in human keratinocytes. The pretreatment with CGM significantly reduced apoptosis in H2O2-exposed HaCaT cells. It promoted the degradation of caspase-3, caspase-8, and caspase-9; and it induced the formation of the processed PARP. The treatment with CGM caused an increase in vesicle formation compared to control group. The level of p62 was reduced and the conversion of LC3-I to LC3-II was increased in CGM treated HaCaT cells. Also, the treatment with CGM increased cleavage of ATG5-ATG12 complex. In summary, CGM helps the cells to survive under stressful conditions by preventing apoptosis and enhancing autophagy. Besides, the present investigation provides evidence to support the antioxidant potential of CGM in vitro and opens up a new horizon for future experiments.

This study was carried out to investigate the effect on the growth and antioxidant activities of Cichorium intybus L.(CLE), Cichorium intybus L. var. folisum ‘treviso’ (CLET), Cichorium intybus L. var. folisum ‘rosaitaliana’ (CLER) in hydroponics added with Cr3+ or Selenium (Se) for 4 weeks. Total polyphenol, total flavonoids contents and FRAP values of three species of chicory were grown hydroponically with Cr3+ or Se were increased. These extracts were also showed stronger DPPH and ABTS scavenging activity than chicory extracts. In particular, chicories added with Cr3+ had higher antioxidant activities than chicories added with Se. CLER and CLE extracts added with Cr3+ were also showed α-glucosidase inhibition activities. These results indicate that chicories were cultivated in culture fluid added with Cr3+ or Se could be used as high functional vegetables.

Nanotechnology has become one of the fastest developing technologies and recently applied to a variety of industries. Thus, increasing number of nano materials including various nanoparticles would be discharged into wastewater and consequently entering a biological wastewater treatment process. However, the impact of the nano particles on biological wastewater treatment has not been estimated intensively. In this research, we investigated the effect of silica nanoparticle on the oxygen uptake rates (OURs) of activated sludge used in a conventional wastewater treatment process. The inhibition (%) values were estimated from the results of OURs experiments for the silica nanoparticles with various sizes of 10-15, 45-50, and 70-100 nm and concentrations of 50, 250, and 500 ppm. As results, the inhibition value was increased as the size of silica nano particles decreased and the injected concentration increased. The maximum inhibition value was investigated as 37.4 % for the silica nanoparticles with the size of 45-50 nm and concentration of 50 ppm. Additionally, the effect of size and concentration on the inhibition should be considered cautiously in case that the aggregation of particles occurred seriously so that the size of individual particles was increased in aquatic solution.

본 연구는 짚신나물 열수 추출물의 α-glucosidase 저해 활 성을 측정하고, 분화된 근육세포에서 glucose 이용과 인슐린 신호전달에 미치는 영향을 분석하였다. 짚신나물 열수 추출 물(10 ㎎/㎖)은 α-glucosidase 활성을 67% 저해하였으며, 같은 농도의 양성대조구인 acarbose(63%)와 유사한 저해 효과를 보였다. 짚신나물 열수 추출물이 α-glucosidase에 의한 단당 류 생성을 저해함으로 식사 후 혈당이 급격히 상승하는 것을 억제하는데 효과적인 소재로 이용 가능성을 확인하였다. 또한 근육세포에서 인슐린 저항성을 유발하기 위해 지방산(1 mM, palmitic acid)를 처리하였고, glucose의 세포내 유입이 감소 되는 것을 확인하였다. 지방산 처리 세포 모델에서 짚신나 물 열수 추출물(10 ㎍/㎖)은 glucose 이용을 유의적으로 회복 시켜 주었다. Normal 상태의 배양조건에서 근육세포의 포도 당 이용능은 짚신나물 열수 추출물(100 ㎍/㎖) 처리에 의해 유의적으로 증가하였다. 근육세포 내로 glucose 유입은 운반 단백질인 Glut4를 통해 이루어지며, 이것은 인슐린이 신호전 달을 통해 조절한다. 짚신나물 열수 추출물의 세포 내 glucose 이용 증가 효과는 인슐린 신호전달 관련 분자인 Akt 유전자 와 단백질 발현을 증가시킨 것과 관련되는 것으로 추정된다. 결론적으로, 짚신나물 열수 추출물은 소화기관에서의 탄수화 물 흡수 저해와 근육세포 내 glucose 이용 증가를 통해 혈당 조절 및 당 대사 개선에 긍정적인 영향을 미치고 있음을 확인 하였다.

Obesity is the most common nutritional disorder in the developed world and has become a global epidemic in recent years. In this study, Zanthoxylum planispinum extracts (ZPE) were evaluated on the effect on inhibition of pancreatic lipase and lipid metabolism by oral treatment for 2 months in high-fed diet obesity-induced Balb/c mice. The ZPE showed pancreatic lipase inhibitory activity with IC50 of 0.3 ㎍/㎖. No significant difference in feed intake was observed among the groups. The high-fat diet-treated Z. planispinum extracts groups (HFD+ZPE, 100 ㎎/㎏) significantly decreased body weight compared to the high-fat diet vehicle groups (HFD, p<0.05). The high-fat diet-treated XenicalⓇ groups (HFD+Xenical, n=10, 30 ㎎/㎏) also showed a significant reduction of body weight compared to HFD (p<0.05). Biochemical parameters (triglyceride, total cholesterol, and high-density lipoprotein cholesterol) in HFD plus ZPE diet groups were significantly lower than those of the HFD groups (p<0.05). These results indicated that ZPE more effectively suppressed the effects of HFD on body fat gain with the inhibitory effect on pancreatic lipase.

The purpose of this study was performed to determine the whitening effect of organic solvent extracts from the centipede, Scolopendra subspinipes mutilans. We prepared different concentrations (50%, 70% and 100%) of ethanol, methanol, 100% ethyl acetate and water extracts. We tested melanin inhibitory effect and tyrosinase activity using B16/F10 melanoma cell. As a result, treatment of organic solvent extracts is decreased the biosynthesis of melanin and tyrosinase activity to 30~60%. Especially the 70% ethanol extracts was the most effective in B16/F10 melanoma cells. In the study on melanogenic protein expression, 70% ethanol extracts of Scolopendra subspinipes mutilans blocked glycosylation of tyrosinase. Therefore this result suggests that 70% ethanol extracts could be developed as a skin whitening agents.

Xylitol is a sugar alcohol with a variety of functions including bactericidal and anticariogenic effects. However, the cellular mechanisms underlying the role of xylitol in bone metabolism are not yet clarified. In our present study, we exploited the physiological role of xylitol on osteoclast dif-ferentiation in a co-culture system of osteoblastic and RAW 264.7 cells. Xylitol treatment of these co-cultures reduced the number of tartrate-resistant acid phosphatase (TRAP)- positive multinucleated cells induced by 10 nM 1α,25(OH)2 D3 in a dose‐dependent manner. A cell viability test revea-led no marked cellular damage by up to 100 mM of xylitol. Exposure of osteoblastic cells to xylitol decreased RANKL, but not OPG, mRNA expression in the presence of 10-8 M 1α,25(OH)2D3 in a dose‐dependent manner. Furthermore, bone resorption activity, assessed on bone slices in the co- culture system, was found to be dramatically decreased with increasing xylitol concentrations. RANKL and OPG proteins were assayed by ELISA and the soluble RANKL (sRANKL) concentration was decreased with an increased xylitol con-centration. In contrast, OPG was unaltered by any xylitol con-centration in this assay. These results indicate that xylitol inhibits 1α,25(OH)2D3-induced osteoclastogenesis by reducing the sRANKL/OPG expression ratio in osteoblastic cells.

Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG 0.2 μM, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.

소화성 질환의 중요 인자인 Helicobacter pylori를 저해하는 IgY, 목이버섯, 김치와 타락 유산균을 이용하여 생육 저해 상승 효과를 분석하고자 하였다. 동결 건조된 유산균 배양 상징액을 다양한 효소 처리 결과, 지질 분해 효소를 제외하고는 활성을 나타냈다. GC 분석을 통해 유산균 동결건조액의 지방산 조성은 undecanoic acid(C11:0), palmitic acid(C16:0), steraic acid(C18:0), oleic acid(C18:1)가 L. mensenteroides LAB KW5와 S. thermophilus LAB KW15에서 모두 확인되었으며, eicosadienoic acid(C20:2)는 LAB KW5에서만 나타났다. 또한 유산균 동결건조액은 다른 식중독균에서도 spot assay의 결과, 그람 음성균 중에서 특히 E. coli O157:H7, E. coli, C. sakazakii 등에서 생육 저해능이 확인되었다. 목이버섯 추출물은 열수 추출과 ethanol를 이용해 분리하였으며, HPLC를 이용하여 목이버섯 추출 다당체를 분석한 결과, glucuronoxylomannan 혹은 glucomannan이 β-glucan과 함께 존재하는 혼합물일 것으로 되었다. 또한, 면역란은 1차 접종 후 11일째부터 주마다 2회씩 회수하여 IgY를 분리, 정제하였다. 실험을 통해 동결 건조된 유산균 배양 상징액, IgY, 목이버섯 추출액을 혼합 배양하여 배양시간에 따른 생육 저해력의 변화를 확인한 결과, 유산균에 의해 H. pylori의 저해 효과가 있었으며, IgY와 목이버섯의 혼합 시 추가 억제 효과가 있는 것으로 나타났다. 그러므로 동결 건조된 유산균 배양 상징액, IgY, 목이버섯 추출액의 복합처리는 H. pylori를 제어하는데 효과적인 것으로 보인다.

Resistance to the induction of apoptosis is a possible me- chanism by which tumor cells can survive anti-neoplastic treatments. Melanoma is notoriously resistant to anti-neop- lastic therapy. Previous studies have demonstrated focal adhesion kinase (FAK) overexpression in melanoma cell lines. Given its probable role in mediating resistance to apo- ptosis, many researchers have sought to determine whether the downregulation of FAK in melanoma cells would confer a greater sensitivity to anti-neoplastic agents. Genistein is a known inhibitor of protein-tyrosine kinase (PTK), which may attenuate the growth of cancer cells by inhibiting the PTK- mediated signaling pathway. This present study was under- taken to investigate the effect of genistein on the expression of FAK and cell cycle related proteins in the G361 me- lanoma cell line. Genistein was found to have a preferential cyto- toxic effect on G361 melanoma cells over HaCaT normal ke- ratinocytes. Genistein decreased the expression of 125 kDa phosphotyrosine kinase and the FAK protein in particular. Genistein treatment did not affect the expression of p53 in G361 cells in which p21 is upregulated. The expression of cy- clin B and cdc2 was downregulated by genistein treatment. Taken together, our data indicate that genistein induces the decreased proliferation of G361 melanoma cells via the in-hibition of FAK expression and regulation of cell cycle genes. This suggests that the use of genistein may be a via- ble approach to future melanoma treatments.

The dual effectiveness of Opuntia ficus indica extracts for browning inhibition and microbial inactivation on fresh-cut apples was investigated. Prepared apple slices were treated with 25, 50, 100, 200 mg/mL Opuntia ficus indica extracts, packaged in polyethylene bags, and stored for 10 days at 4, 21oC. Results indicate that Opuntia ficus indica extracts significantly (P < 0.05) inhibited the browning reaction of fresh-cut apples. This treatment also reduced peroxidase activities. The populations of Staphylococcus aureus significantly decreased with increasing extract concentration (p < 0.05). In particular, S. aureus was reduced to non-detectable levels after 2 days in 100 mg/mL treatment at 4oC and 21oC. Opuntia ficus indica extracts therefore have antibacterial and antibrowning effects. The results suggest that Opuntia ficus indica extracts could be useful as a natural food preservative.

The purpose of this study was to examine the anti-metastatic effect of Juniperus chinensis extract by inhibition of galectin-3 expression. For this study, we carried out the experiment of galectin-3 expression assay and in vivo metastasis mouse model bearing colon 26-M3.1 lung carcinoma, and also MMP-1, MMP-2 and MMP-9 inhibitory assays relating metastasis mechanism. J. chinensis extract significantly showed the inhibitory effect of basal expression and MNNG-induced expression of galectin-3 in colon 26-M3.1 lung carcinoma at the all tested doses (5, 50 and 500 ㎍/㎖). In case of the inhibition of MMP-1, MMP-2 and MMP-9, the weak activity was showed at the doses of 50 and 500 ㎍/㎖ and was not detected at the dose of 5 ㎍/㎖. However, J. chinensis extract significantly showed the anti metastatic activity on mouse mode bearing colon 26-M3.1 lung carcinoma. Therefore, this study strongly suggests that J. chinensis is a potent inhibitor of galectin-3 expression, and holds great promise for use in inhibiting metastasis induced by elevated expression of galectin-3.

영지버섯 자실체를 열수추출과 주정추출을 하여 항산화 효능 및 간암 및 위암세포의 생장 저해도를 분석하였다. 항산화 효능을 실시한 결과, 대조군인 Trolox, BHA보다 높은 항산화 효능을 보인 것은 ASI 7004, 7014이며, 이 두 균주는 열수추출물과 주정 추출물에서 모두 높은 항산화 효능을 나타냈다. 나머지 균주는 대조군인 ABTs보다 대체적으로 높은 효능을 보였다. 또한 암세포 생장저해도를 알아보기 위해서 열수, 주정 추출물을 간암세포인 HepG2에 농도별로(100, 200, 400μg/ml) 처리하여, 세포 생장 저해도(MTT assay)를 측정한 결과, 열수 추출물에서는 ASI 7002, 7011, 7014, 7020이 농도 의존적으로 간암세포의 생장을 저해하는 것을 알 수 있었다. 또한 주정 추출물에서는 ASI 7011, 7019가 농도 의존적으로 간암세포 생장을 저해하는 것으로 나타났다. 위암세포인 AGS에 농도별로(100, 200, 400μg/ml)처리하여, 세포 생장 저해도(MTT assay)를 측정한 결과, 열수 추출물에서는 ASI 7001, 7002, 7019, 7020이 농도 의존적으로 위암세포의 생장을 저해하는 것을 알 수 있었다. 또한 주정 추출물에서는 ASI 7001, 7002가 농도 의존적으로 위암세포 생장을 저해하는 것으로 나타났다.

The pinewood nematode, Bursaphelenchus xylophilus is a serious forest pathogen in many countries including Japan, China and Korea. To minimize the environmental problems caused by synthetic chemicals broadly utilized in the control of B. xylophilus, we estimated the nematicidal potency of 63 aliphatic compounds against B. xylophilus by measuring their inhibition activity against acetylcholinesterases (ACE, EC 3.1.1.7) of B. xylophilus (BxACEs). In the primary inhibition assay using B. xylophilus crude protein, C6, C9, C10 2E-alkenal C12 alkanoic acid were observed the > 45% BxACE inhibition rate and they were subsequently estimated the inhibition rate against three recombinant BxACEs. Whole compounds showed the high inhibition rate against BxACE-1 and BxACE-2. Interestingly, C6 2E-alkenal and C12 alkanoic acid exhibited the high inhibition rate against BxACE-3 which would be insensitive to ACE inhibitors.

This study was conducted to examine the optimal concentration and treatment time of antioxidants for inhibition of the ROS generation in bovine somatic cell nuclear transfer (SCNT) embryos. Bovine oocytes were activated parthenogenetically, during which oocytes were treated with various antioxidants to determine the optimal concentrations and kind of antioxidants. Determined antioxidants were applied to oocytes during in vitro maturation (IVM) and/or SCNT procedures. Finally, antioxidant-treated SCNT embryos were compared with in vitro fertilized (IVF) embryos. H2O2 levels were analyzed in embryos at 20 h of activation, fusion or insemination by staining of embryos in 10 μM 2'7'-dichlorodihydrofluorescein diacetate (H2DCFDA) dye, followed by fluorescence microscopy. H2O2 levels of parthenogenetic embryos were significantly lower in 25 μM β- mercaptoethanol (β-ME), 50 μM L-ascorbic acid (Vit. C), and 50 μM L-glutathione (GSH) treatment groups than each control group (24.0±1.5 vs 39.0±1.1, 29.7±1.0 vs 37.0±1.2, and 32.9±0.8 vs 36.3±0.8 pixels/embryo, p<0.05). There were no differences among above concentration of antioxidants in direct comparison (33.6±0.9～35.2±1.1 pixels/embryo). Thus, an antioxidant of 50 μM Vit. C was selected for SCNT. H2O2 levels of bovine SCNT embryos were significantly lower in embryos treated with Vit. C during only SCNT procedure (26.4±1.1 pixels/embryo, p<0.05) than the treatment group during IVM (29.9±1.1 pixels/embryo) and non-treated control (34.3±1.0 pixels/embryo). Moreover, H2O2 level of SCNT embryos treated with Vit. C during SCNT procedure was similar to that of IVF embryos. These results suggest that the antioxidant treatment during SCNT procedures can reduce the ROS generation level of SCNT bovine embryos.