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Inhibition of Migration and Invasion of LNCap Human Prostate Carcinoma Cells by Doxorubicin through Inhibition of Matrix Metalloproteinase Activity and Tightening of Tight Junctions (Doxorubicin에 의한 치밀결합 강화 및 MMPs의 활성 억제를 통한 LNCap 전립선 암세포의 이동성 및 침윤성의 억제)

  • Choi, Yung Hyun;Shin, Dong Yeok;Kim, Wun-Jae
    • Journal of Life Science
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    • v.24 no.6
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    • pp.700-706
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    • 2014
  • Doxorubicin (trade name adriamycin), an anthracycline antibiotic, is commonly used in the treatment of a wide range of cancers, including hematological malignancies, many types of carcinoma, and soft tissue sarcomas. It is closely related to the natural product daunomycin, and like all anthracyclines, it works by intercalating DNA. Its most serious adverse effect is life-threatening heart damage. Its anti-metastatic mechanisms in human prostate carcinomas are not fully understood. In this study, we used LNCap human prostate carcinoma cells to investigate the inhibitory effects of doxorubicin on cell motility and invasion, two critical cellular processes that are often deregulated during metastasis. Doxorubicin treatment inhibited cell migration and invasiveness of LNCap cells without showing any toxicity. Doxorubicin treatment also suppressed the activity and expression of matrix metalloproteinase (MMP)-2 and MMP-9, which were associated with up-regulated expression of tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 in LNCap cells. Doxorubicin treatment also attenuated the expression levels of claudin family members (claudin-1, -2,-3 and -4), major components of tightening of tight junctions (TJs) and increased the tightening of TJs, as demonstrated by an increase in transepithelial electrical resistance. The present findings demonstrate that doxorubicin reduces the migration and invasion of prostate carcinomas LNCap cells by modulating the activity of TJs and MMPs.

Establishment of Microwave-Assisted Extraction Conditions for Antioxidative Extracts from Pleurotus eryngii (새송이버섯의 항산화성 추출물 제조를 위한 마이크로웨이브 추출조건 설정)

  • Oh, Hyun-In;Lim, Tae-Soo;Lee, Gee-Dong;Kim, Hyun-Ku
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.1-6
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    • 2006
  • Through the application of a microwave-assisted extraction, the aim of study was to investigate the extraction conditions for the total yield, electron donating ability, total polyphenol contents and nitrite scavenging effect from Pleurotus eryngii. This experiment was conducted under the following extraction conditions: the microwave power (60, 90, 120 W), sample to solvent ratio (1:50, 1:20, 1:10), extraction time (1, 5, 10, 15 mins), and extractant like water, $50\%$ EtOH and $99\%$ EtOH. During the increase of microwave power ranging from 60 to 120 W, the maximized effective components were obtained when the microwave power was set at 60 W. By decreasing the ratio down to 1:10 (g/mL), the extraction of effective components but not total yield was the highest at 1:10 (g/mL). As going up to 15 min, the maximum extraction of effective components was achieved at 1 min for the extraction time and was little affected at over 5 min. In conclusion, the most appropriate conditions for the extraction of effective compounds from Pleurotus eryngii were as follows: the microwave power at 60 W, 1:10 of the sample to solvent ratio, extraction time at 1 min and water as the extractant.

Risk Assessment Tools for Invasive Alien Species in Japan and Europe (일본과 유럽의 침입외래생물 생태계위해성평가 기법)

  • Kil, Jihyon;Mun, Saeromi;Kim, Chang-Gi
    • Ecology and Resilient Infrastructure
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    • v.2 no.3
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    • pp.191-197
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    • 2015
  • Invasive alien species are considered to be one of the main factors that cause biodiversity loss. Establishment of management strategies through continuous monitoring and risk assessment is a key element for invasive alien species management policy. In the present study, we introduce examples of ecological risk assessment tools developed in Japan, Germany-Austria and Belgium. Invasive alien species have been designated in Japan based on the assessment of risks to ecosystems, human health and primary industry. German-Austrian Black List Information System categorized alien species into Black List, White List and Grey List according to their risks to biodiversity. In the Harmonia Information System developed in Belgium, invasiveness, adverse impacts on native species and ecosystem functions and invasion stages were assessed and alien species were categorized into Black List, Watch List and Alert List. These international risk assessment tools may be helpful to improve our national risk assessment protocol for the prioritization of invasive alien species management.

Effect of Mulching and Soil Conditioners on Yield and Flavonoids Content of Mungbean (녹두 수량과 Flavonoids 함량에 대한 멀칭과 토양개량제 효과)

  • Kim, Dong-Kwan;Chon, Sang-Uk;Lee, Kyung-Dong;Son, Dong-Mo;Rim, Yo-Sup;Kim, Kyong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.4
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    • pp.353-358
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    • 2008
  • The purpose of this study was to examine the effects of black polyethylene film mulching and soil conditioners on the growth, yield, and flavonoids biosynthesis of mungbeans. The seedling stand rate, plant height, and pods per plant were 98%, 61 cm, and 15.1, respectively, when the mungbean grown black polyethylene film mulches. They were greater than these results when the without mulches was applied by 13%, 9 cm, and 1.8, respectively. Mungbean grown with black polyethylene film mulches ($243\;kg{\cdot}10a^{-1}$) exhibited 38% more seed yield than without mulches ($176\;kg{\cdot}10a^{-1}$). However, no significant difference in contents of vitexin and isovitexin of mungbeans between with black polyethylene film mulches and without mulches was observed. Among soil conditioners, standard fertilizer+oil cake and standard fertilizer+mineral powders increased number of pods. The standard fertilizer+oil cake showed the highest seed yield (119%) of mungbean when it mixed with standard fertilizers, and followed by mineral powders (115%). The mixed combinations of oil cake or mineral powders also seem to have affected the biosynthesis of vitexin and isovitexin, whose content in mungbeans was significantly more.

Thin Layer Drying Model of Sorghum

  • Kim, Hong-Sik;Kim, Oui-Woung;Kim, Hoon;Lee, Hyo-Jai;Han, Jae-Woong
    • Journal of Biosystems Engineering
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    • v.41 no.4
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    • pp.357-364
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    • 2016
  • Purpose: This study was performed to define the drying characteristics of sorghum by developing thin layer drying equations and evaluating various grain drying equations. Thin layer drying equations lay the foundation characteristics to establish the thick layer drying equations, which can be adopted to determine the design conditions for an agricultural dryer. Methods: The drying rate of sorghum was measured under three levels of drying temperature ($40^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$) and relative humidity (30%, 40%, and 50%) to analyze the drying process and investigate the drying conditions. The drying experiment was performed until the weight of sorghum became constant. The experimental constants of four thin layer drying models were determined by developing a non-linear regression model along with the drying experiment results. Result: The half response time (moisture ratio = 0.5) of drying, which is an index of the drying rate, was increased as the drying temperature was high and relative humidity was low. When the drying temperature was $40^{\circ}C$ at a relative humidity (RH) of 50%, the maximum half response time of drying was 2.8 h. Contrastingly, the maximum half response time of drying was 1.2 h when the drying temperature was $60^{\circ}C$ at 30% RH. The coefficient of determination for the Lewis model, simplified diffusion model, Page model, and Thompson model was respectively 0.9976, 0.9977, 0.9340, and 0.9783. The Lewis model and the simplified diffusion model satisfied the drying conditions by showing the average coefficient of determination of the experimental constants and predicted values of the model as 0.9976 and Root Mean Square Error (RMSE) of 0.0236. Conclusion: The simplified diffusion model was the most suitable for every drying condition of drying temperature and relative humidity, and the model for the thin layer drying is expected to be useful to develop the thick layer drying model.

GABA-enriched fermented Laminaria japonica improves cognitive impairment and neuroplasticity in scopolamine- and ethanol-induced dementia model mice

  • Reid, Storm N.S.;Ryu, Je-kwang;Kim, Yunsook;Jeon, Byeong Hwan
    • Nutrition Research and Practice
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    • v.12 no.3
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    • pp.199-207
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    • 2018
  • BACKGROUND/OBJECTIVES: Fermented Laminaria japonica (FL), a type sea tangle used as a functional food ingredient, has been reported to possess cognitive improving properties that may aid in the treatment of common neurodegenerative disorders, such as dementia. MATERIALS/METHODS: We examined the effects of FL on scopolamine (Sco)- and ethanol (EtOH)-induced hippocampus-dependent memory impairment, using the Passive avoidance (PA) and Morris water maze (MWM) tests. To examine the underlying mechanisms associated with neuroprotective effects, we analyzed acetylcholine (ACh) and acetylcholinesterase (AChE) activity, brain tissue expression of muscarinic acetylcholine receptor (mAChR), cAMP response element binding protein (CREB) and extracellular signal-regulated kinases 1/2 (ERK1/2), and immunohistochemical analysis, in the hippocampus of mice, compared to current drug therapy intervention. Biochemical blood analysis was carried out to determine the effects of FL on alanine transaminase (ALT), aspartate transaminase (AST), and triglyceride (TG) and total cholesterol (TC) levels. 7 groups (n = 10) consisted of a control (CON), 3 Sco-induced dementia and 3 EtOH-induced dementia groups, with both dementia group types containing an untreated group (Sco and EtOH); a positive control, orally administered donepezil (Dpz) (4mg/kg) (Sco + Dpz and EtOH + Dpz); and an FL (50 mg/kg) treatment group (Sco + FL50 and EtOH + FL50), orally administered over the 4-week experimental period. RESULTS: FL50 significantly reduced EtOH-induced increase in AST and ALT levels. FL50 treatment reduced EtOH-impaired step-through latency time in the PA test, and Sco- and EtOH-induced dementia escape latency times in the MWM test. Moreover, anticholinergic effects of Sco and EtOH on the brain were reversed by FL50, through the attenuation of AChE activity and elevation of ACh concentration. FL50 elevated ERK1/2 protein expression and increased p-CREB (ser133) in hippocampus brain tissue, according to Western blot and immunohistochemistry analysis, respectively. CONCLUSION: Overall, these results suggest that FL may be considered an efficacious intervention for Sco- and EtOH-induced dementia, in terms of reversing cognitive impairment and neuroplastic dysfunction.

Immunotoxicity Study of Separated Antigen from Helicobacter pylori. (Helicobacter pylori로부터 유래된 항원의 항원성에 관한 연구)

  • Park, Chang-Ho;Bae, Man-Jong
    • Journal of Life Science
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    • v.18 no.4
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    • pp.494-502
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    • 2008
  • The anaphylaxis shock reaction on the whole cells of H. pylori exhibited a symptom of slight illness for the first and second medication of causing antigen at an antigen concentration of WC (H) $60\;{\mu}g/100\;{\mu}l$ for WC (H) and no anaphylaxis shock symptom was observed at an antigen concentration of $20\;{\mu}g/100\;{\mu}l$ for WC (L). In the case of anaphylaxis shock reaction on the crude urease, no symptom was observed at an antigen concentration of $20\;{\mu}g/100\;{\mu}l$ for both urease (L) and urease (H). In the heterologous passive cutaneous anaphylaxis (PCA) test using a guinea pig-rat, no positive reaction was detected in all the medication groups of WC (H), WC (L), urease (H) and urease (L). In the skin sensitization test, it was observed that the best antigen concentration not causing skin disorder at each of $80\;{\mu}g/100\;{\mu}l$, $40\;{\mu}g/100\;{\mu}l$, $20\;{\mu}g/100\;{\mu}l$, and $20\;{\mu}g/100\;{\mu}l$ was $40\;{\mu}g/100\;{\mu}l$.

Effects of (-)-Epigallocatechin-3-gallate on the Release of Pancreatic Enzymes and Expression of Regenerating Genes in Ethanol-injured Murine Pancreatic Primary Acinar Cells (에탄올에 의하여 유도된 마우스 췌장 선포세포의 염증성 손상에서 췌장분비 효소의 활성 및 세포 재생관련 유전자들의 발현에 미치는 EGCG의 영향)

  • Kim, Sung Ok;Choi, Yung Hyun
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1404-1408
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    • 2013
  • (-)-Epigallocatechin-3-gallate (EGCG), a green tea polyphenol, has been shown to have strong antibacterial, antiviral, antioxidant, anti-inflammatory, and chemopreventive effects. However it is unknown whether EGCG can recover alcohol-associated pancreatitis. The aim of this study was to investigate the effects of EGCG on pancreatic enzyme activities and the expressions of pancreatic regenerating related markers, such as adenosine monophosphate-activated protein kinase (AMPK), raf-1 kinase inhibitor protein (RKIP), and Regenerating gene 1 (Reg1), in mice pancreatic primary acinar cells. Our results revealed that activities of ${\alpha}$-amylase and chymotrypsin were significantly increased in the cells treated with ethanol compared to the untreated control cells; however, the increased activities of both enzymes were markedly reduced by pretreatment with EGCG. Phosphorylation of AMPK and total expression of RKIP were decreased in the ethanol-treated primary acinar cells; however, these were both significantly increased in the EGCG-pretreated cells. In addition, when EGCG was treated, expression of Reg1 was markedly increased compared with that of the control or the ethanol-treated primary acinar cells, demonstrating that EGCG can modulate pancreatic regenerating related genes. Therefore, our findings suggest that EGCG may have therapeutic utility in the prevention or treatment of alcohol-associated pancreatitis.

Zinc deficiency decreased cell viability both in endothelial EA.hy926 cells and mouse aortic culture ex vivo and its implication for anti-atherosclerosis

  • Cho, Young-Eun;Choi, Jee-Eun;Alam, Md. Jahangir;Lee, Man-Hyo;Sohn, Ho-Yong;Beattie, John H.;Kwun, In-Sook
    • Nutrition Research and Practice
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    • v.2 no.2
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    • pp.74-79
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    • 2008
  • Zinc plays a protective role in anti-atherosclerosis but the clear mechanism has not been proposed yet. In the present study, we evaluated whether zinc modulates atherosclerotic markers, VACM-1 and ICAM-1 and cell viability both in endothelial cells in vitro and mouse aortic cell viability ex vivo. In study 1, as in vitro model, endothelial EA.hy926 cells were treated with $TNF{\alpha}$ for 5 hours for inducing oxidative stress, and then treated with Zn-adequacy ($15\;{\mu}M$ Zn) or Zn-deficiency ($0\;{\mu}M$ Zn) for 6 hours. Pro-atherosclerosis factors, VCAM-1 and ICAM-1 mRNA expression and cell viability was measured. In study 2, as ex vivo model, mouse aorta ring was used. Mourse aorta was removed and cut in ring then, cultured in a 96-well plate. Aortic ring was treated with various $TNF{\alpha}$ (0-30 mg/ml) and intracellular zinc chelator, N, N, N', N', -tetrakis (2-pyridylmethyl) ethylenediamine (TPEN, $0-30\;{\mu}M$) for cellular zinc depletion for 2 days and then cell viability was measured. The results showed that in in vitro study, Zn-adequate group induced more VCAM-1 & ICAM-1 mRNA expression than Zn-deficient group during 6-hour zinc treatment post-5 hour TNF-$\alpha$ treatment, unexpectedly. These results might be cautiously interpreted that zinc would biologically induce the early expression of anti-oxidative stress through the increased adhesion molecule expression for reducing atherosclerotic action, particularly under the present 6-hour zinc treatment. In ex vivo, mouse aortic ring cell viability was decreased as TNF-$\alpha$ and TPEN levels increased, which suggests that mouse aortic blood vessel cell viability was decreased, when oxidative stress increases and cellular zinc level decreases. Taken together, it can be suggested that zinc may have a protective role in anti-atherosclerosis by cell viability in endothelial cells and aorta tissue. Further study is needed to clarify how pro-atherosclerosis molecule expression is modulated by zinc.

Increased Production of an Alkaline Protease from Bacillus clausii I-52 by Chromosomal Integration (Bacillus clausii I-52의 Chromosomal Integration에 의한 Alkaline Protease의 생산성 향상)

  • Joo, Han-Seung;Park, Dong-Chul;Choi, Jang-Won
    • Journal of agriculture & life science
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    • v.46 no.1
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    • pp.163-176
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    • 2012
  • TTo increase productivity of a strong extracellular alkaline protease (BCAP), stable strains of Bacillus clausii I-52 carrying another copy of BCAP gene in the chromosome were developed. Integrative vector, pHPS9-fuBCAP carrying BCAP promoter, ribosome binding site, signal sequence and active protease gene was constructed and transferred into B. clausii I-52, and integration of the constructed plasmid into chromosome was identified by PCR. An investigation was carried out on BCAP production by B. clausii I-52 and transformant C5 showing the highest relative activity of alkaline protease using submerged fermentation. Maximum enzyme activity was produced when cells were grown under the submerged fermentation conditions at $37^{\circ}C$ for 48 h with an aeration rate of 1 vvm and agitation rate of 650 rpm in a optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_47H_2O$ 0.01%, $FeSO_47H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). A protease yield of approximately 134,670U/ml was achieved using an optimized media, which show an increase of approximately 1.6-fold compared to that of non-transformant (83,960 U/ml). When the stability of transformant C5 was examined, the integrated plasmid pHPS9-fuBCAP was detected in the transformant after cultivation for 8 days, suggesting that it maintained stably in the chromosomal DNA of transformant C5.