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Anti-metastatic Effects on B16F10 Melanoma Cells of Extracts and Two Prenylated Xanthones Isolated from Maclura amboinensis Bl. Roots

  • Siripong, Pongpun;Rassamee, Kitiya;Piyaviriyakul, Suratsawadee;Yahuafai, Jantana;Kanokmedhakul, Kwanjai
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3519-3528
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    • 2012
  • Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.

MicroRNA-101 Inhibits Cell Proliferation, Invasion, and Promotes Apoptosis by Regulating Cyclooxygenase-2 in Hela Cervical Carcinoma Cells

  • Huang, Fei;Lin, Chen;Shi, Yong-Hua;Kuerban, Gulinar
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5915-5920
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    • 2013
  • Aim: Although aberrant miRNA expression has been documented, altered miR-101 expression in cervical cancer and its carcinogenic effects and mechanisms remain unexplored. The aim of our study was to investigate the role of miR-101 alteration in cervical carcinogenesis. Methods: Expression of miR-101 was examined by quantitative real-time reverse transcriptase PCR (qRT-PCR) in Hela cells. After modulating miR-101 expression using miR-101 mimics, cell growth, apoptosis and proliferation, and migration were tested separately by MTT or flow cytometry and cell wound healing assay and protein expression was detected by qRT-PCR. The expression of COX-2 in Hela cell was also examined by immunohistochemical staining and the correlation with miR-101 expression was analysed. Results: The miR-101 demonstrated significantly low expression in Hela cell. When we transfected miR-101 mimics into Hela cells, the modulation of miR-101 expression remarkably influenced cell proliferation, cycling and apoptosis: 1) The expression of microRNA-101 tended to increase after transfection; 2) Overexpression of miR-101 was able to promote cell apoptosis, the apoptosis rate being markedly higher (97.6%) than that seen pre-transfection (12.2%) (P<0.05); 3) The miR-101 negatively regulates cell migration and invasion, scratch results being lower ($42.7um{\pm}2um$) than that observed pre-transfection ($181.4um{\pm}2um$); 4) miRNA-101 inhibits the proliferation of Hela cells as well as the level of COX-2 protein, which was negatively correlated with miR-101 expression. Conclusions: Overexpression of miR-101 has obvious inhibitory effects on cell proliferation, migration and invasion. Thus reduced miR-101 expression could participate in the development of cervical cancer at least partly through loss of inhibition of target gene COX-2, which probably occurs in a relative late phase of carcinogenesis. Our data suggest an important role of miR-101 in the molecular etiology of cancer and indicate potential application of miR-101 in cancer therapy.

The Influence of Water Temperature and Body Weight on Metabolic Rate of Olive Flounder Paralichthys olivaceus (넙치 Paralichthys olivaceus 대사율에 미치는 수온과 체중의 영향)

  • Oh, Sung-Yong;Jang, Yo-Soon;Park, Heung-Sik;Choi, Young-Ung;Kim, Chong-Kwan
    • Ocean and Polar Research
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    • v.34 no.1
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    • pp.93-99
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    • 2012
  • The effect of water temperature and body weight on oxygen consumption by the fasted olive flounder Paralichthys olivaceus was investigated in order to assess the metabolic rate of this species under different conditions. The oxygen consumption rate (OCR) was measured at three different water temperatures (15, 20 and $25^{\circ}C$) and two different body weights [$9.1{\pm}1.2$ g (mean${\pm}$SD) for the juvenile group and $266.4{\pm}29.3$ g for the immature group] at an interval of 5 minutes for 24 hours using a closed flow-through respirometer. For each treatment condition, three replicates were set up and 135 fish in the juvenile group and 18 fish in the immature group were used. The OCRs exhibited a linear increase described by OCR=-82.06+28.30T ($r^2$=0.96, p<0.001) in the juvenile group and OCR=-52.52+14.73T ($r^2$=0.97, p<0.001) in the immature group. The OCRs decreased with increasing body weights at a given water temperature (p<0.001). The metabolic rate was related to the body weight of the fish as a power function with a weight exponent of between 0.77 and 0.82. $Q_{10}$ values ranged 1.67~2.28 when the temperature was between 15 and $20^{\circ}C$, 1.57~1.93 when the temperature was between 20 and $250^{\circ}C$, and 1.79~1.89 when the temperature was between 15 and $250^{\circ}C$. The energy expenditure by respiration increased with increasing water temperature and decreasing body weight (p<0.001). The mean energy loss rates at 15, 20 and $25^{\circ}C$ were 115.9, 149.8 and 208.2 kJ $kg^{-1}d^{-1}$ in the juvenile groups and 53.8, 81.2 and 101.9 kJ $kg^{-1}d^{-1}$ in the immature groups.

Inhibition of PI3K/AKT Signaling Pathway Enhances Cordycepin-Induced Apoptosis in Human Gastric Cancer Cells (인체위암 세포에서 PI3K/AKT 신호 전달계 차단에 의한 동충하초 유래 Cordycepin의 Apoptosis 유발 효과 증진)

  • Lee, Hye Hyeon;Jeong, Jin-Woo;Choi, Yung Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.6
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    • pp.835-842
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    • 2016
  • The phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway plays a crucial role in cancer occurrence by promoting cell proliferation and inhibiting apoptosis. In the present study, we evaluated the effect of a PI3K inhibitor, LY294002, on the chemosensitivity of gastric cancer cells to cordycepin, a predominant functional component of the fungus Cordyceps militaris, in AGS human gastric cancer cells and investigated possible underlying cellular mechanisms. Our results revealed that cordycepin inhibited viability of AGS cells in a concentration-dependent manner and induced apoptosis, as determined by apoptotic cell morphologies and fluorescence-activated cell sorting analysis associated with attenuated activation of the PI3K/Akt signaling pathway. Treatment with cordycepin in combination with a subtoxic concentration of LY294002 enhanced cordycepin-induced cytotoxicity and apoptotic potentials in AGS cells. Sensitization of LY294002 to cordycepin-induced apoptosis was accompanied by activation of caspases (caspases-3, -8, and -9) and was concomitant with poly(ADP-ribose) polymerase cleavage. Moreover, LY294002 up-regulated pro-apoptotic Bax and enhanced truncation of Bid in cordycepin-treated AGS cells, which was connected with increased loss of mitochondrial membrane potential and release of cytochrome c from mitochondria to the cytosol. Taken together, these results indicate that inhibition of the PI3K/Akt signaling pathway could augment cordycepin-induced apoptosis in human gastric cancer cells by up-regulating caspase activity through mitochondrial dysfunction.

Comparison of meat quality, fatty acid composition and aroma volatiles of dry-aged beef from Hanwoo cows slaughtered at 60 or 80 months old

  • Utama, Dicky Tri;Kim, Yeong Jong;Jeong, Hae Seong;Kim, Juntae;Barido, Farouq Heidar;Lee, Sung Ki
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.1
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    • pp.157-165
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    • 2020
  • Objective: The objective of this study was to compare the quality of dry-aged beef from cull Hanwoo cows slaughtered at 60 or 80 months old. Methods: A total of eight cull Hanwoo carcasses with a quality grade of 3 (low-grade) were selected and divided into two age groups: 63.5±2.5 months old (n = 4) and 87.8±4.5 months old (n = 4). Whole longissimus thoracis et lumborum from the 11th rib to the last lumbar vertebrae, including the back fat, was removed from the carcass at 24 h postmortem and aged for 50 days in darkness at a temperature of 2℃±1℃, a relative humidity of 85% and an air flow of 2 m/s. The sampling was performed aseptically after 0, 20, 24, 40, and 50 days of aging. Results: Regardless of the aging period, aging increased the lightness (p<0.05), redness (p<0.05) and yellowness (p<0.05) at initial blooming (90 min after slicing) and the overall acceptance (p<0.05). No further tenderization effect was found after 20 days of aging, but aging for 50 days significantly increased the lipid oxidation (p<0.05). The generation of aroma volatiles in the roast steak from aged samples was higher (p<0.05) than that of non-aged samples. No significant effect of age at slaughter was found on the color, pH, water-holding capacity, cooking loss, shear force value, bacterial counts, volatile basic nitrogen, consumer acceptance, lipid oxidation, fatty acid composition or aroma volatiles. Conclusion: The quality of dry-aged beef obtained from cull Hanwoo cows slaughtered at either 60 or 80 months old with similar quality grade was comparable and extending dry aging for more than 40 days is not recommended considering the costs and further lipid oxidation.

An Algorithm for Submarine Passive Sonar Simulator (잠수함 수동소나 시뮬레이터 알고리즘)

  • Jung, Young-Cheol;Kim, Byoung-Uk;An, Sang-Kyum;Seong, Woo-Jae;Lee, Keun-Hwa;Hahn, Joo-Young
    • The Journal of the Acoustical Society of Korea
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    • v.32 no.6
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    • pp.472-483
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    • 2013
  • Actual maritime exercise for improving the capability of submarine sonar operator leads to a lot of cost and constraints. Sonar simulator maximizes the capability of sonar operator and training effect by solving these problems and simulating a realistic battlefield environment. In this study, a passive sonar simulator algorithm is suggested, where the simulator is divided into three modules: maneuvering module, noise source module, and sound propagation module. Maneuvering module is implemented in three-dimensional coordinate system and time interval is set as the rate of vessel changing course. Noise source module consists of target noise, ocean ambient noise, and self noise. Target noise is divided into modulated/unmodulated and narrowband/broadband signals as their frequency characteristics, and they are applied to ship radiated noise level depending on the vessel tonnage and velocity. Ocean ambient noise is simulated depending on the wind noise considering the waveguide effect and other ambient noise. Self noise is also simulated for flow noise and insertion loss of sonar-dome. The sound propagation module is based on ray propagation, where summation of amplitude, phase, and time delay for each eigen-ray is multiplied by target noise in the frequency domain. Finally, simulated results based on various scenarios are in good agreement with generated noise in the real ocean.

A Study on the Feasibility Improvement of the Real Estate Development by Using Project Financing Analytical Method in Korea (PF대출 분석기법을 활용한 부동산개발사업 사업성 평가 개선 연구)

  • Seo, Jeong-Jin
    • Journal of Cadastre & Land InformatiX
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    • v.44 no.2
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    • pp.209-230
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    • 2014
  • There are three forms of REITs company in Korea that was first introduced in 2002. Each REITs have been listed on the KRX, its characteristics are different, but it is classified as a REITs company in all events. REITs current methods are applied uniformly manner that does not reflect the characteristics of the individual. REITs some, that is not seen unlike legislative intent, it is delisted, such as generating an investment loss of investors. In this study it is an object of the invention from the point of view of REITs business validity, to draw up operational support aggressive plans of the scheme. By improving the PF assesment system, to improve the relevance of REIT business and presenting policy direction to the activation of REITs. Through the sophistication of real estate finance utilizing REITs, policy for proper investment of general investors REITs funds were listed with the smooth flow must be realized. The results of this study, it can be utilized as basic data for policy to reflect the real estate policy for activation of the indirect financial investments.

Chemical Neurolytic Block with Absolute Ethyl Alcohol on Cervical Sympathetic Ganglion in Rabbits (토끼에서 경부 교감신경절의 무수 에틸 알코올에 의한 화학적 차단)

  • Kang, Yoo-Jin;Suh, Jae-Hyun
    • The Korean Journal of Pain
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    • v.7 no.2
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    • pp.162-169
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    • 1994
  • Blockade of cervicothoracic sympathetic ganglion (stellate ganglion controls pain on face, head, neck, shoulder, upper limbs, and upper chest, including their viscera and sympathetically maintained pain. This procedure also increases blood flow to the above areas and relieves hyperreactivity of sympathetic nervous system. Clinically, repeated stellate ganglion blocks with local anesthetic agent may become difficult with complications such as accidental intravascular or subdural injection, recurrent laryngeal nerve or bracheal plexus paralysis, pneumothorax and edema on injection site. Therefore, at times long-term cervicothoracic ganglion block with neurolytics is necessitated but its applications are prohibited by the critical structures surrounding ganglion. There are also few reports of neurolytic stellate ganglion block. This study was performed to observe the complications, gross changes of surrounding structures, and microscopic findings of ganglion cells after neurolytic block and to certify the possibility of clinical use of neruolytic stellate ganglion block. The unilateral superior cervical sympathetic ganglion of rabbit was blocked with absolute ethyl alcohol 0.4 ml at the level of cricoid cartilage. Normal ganglion was used as a control and 5 animals were sacrificed at each intervals of 7, 15 and 50 days after block. The results were as follows; 1) All experimental animals showed no specific changes of behavior, motor function. No necrotic tissues were present in the block area during the observation period. There were some gross scar tissues along the fascia of muscles surrounding the needle injection site, but gross atrophy of muscles or injured major vessels were not found. 2) Microscopically, structures of normal ganglion of rabbit were very similar to those of humans. Seven days after absolute ethyl achohol injection there were marked edema of ganglion cells and nuclei with irregular nuclear membrane. Some of the ganglion cells lost their nuclei and showed degenerative changes. Fifteen days after block, cell edema were decreased and loss of the Nissl's body was prominant. The ganglion cell structures looked close to normal but the cytoplasm and nucleus were generally contracted 50 days after block. These results suggest absolute ethyl alcohol injection on cervical sympathetic ganglion with above method mainly blocks pre- and post-synaptic fibers and the long-term neurolytic blockade of this ganglion may be possible in rabbits.

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Efficient Gene Delivery into Hematopoietic Stem Cells by Intra-Bone Marrow Injection of Retrovirus (IBM 이식을 통한 골수 조혈 줄기 세포에의 효과적인 유전자 도입)

  • Lee, Byun-Joo;Lee, Yong-Soo;Kim, Hye-Sun;Kim, Yu-Kyung;Kim, Jae-Hwan;Park, Jin-Ki;Chung, Hak-Jae;Chang, Won-Kyong;Kim, Dong-Ku
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.9-14
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    • 2008
  • Efficient gene transfer into hematopoietic stem cells is a great tool for gene therapy of hematopoietic disease. Retrovirus have been extensively used for gene delivery and gene therapy. However, current in vitro gene transfer has some obstacles suck as induction of differentiation loss of self-renewal capacity, and down-regulation of homing efficiency for in vitro hematopoietic stem cells transplantation. To overcome these problems, we developed efficient in vitro retroviral transfer technique by direct intra-bone marrow injection (IBM). We identified effective retrovirus gene transfer in bone marrow hematopoietic cells in vitro. Two weeks after retrovirus transfer via IBM injection, we observed stable EGFP gene expression in bone marrow, lymph node, spleen, and liver cells. In addition, $6.4{\pm}2.7%$ of hematopoietic stem/progenitor cells were expressed EGFP transgene from flow cytometry analysis. Our results demonstrate that in vitro retrovirus gene transfer via IBM injection can provide a viable alternative to current or moo gene transfer approach.

Nephropathy in Chronic Lead Poisoning (만성 연중독자에서 발생한 신장해)

  • Kim, Byoung-Gwon;Kim, Sung-Ryul;Hong, Young-Seoub;Rha, Seo-Hee;Kim, Jung-Man;Jung, Kap-Yull;Kim, Joon-Youn
    • Journal of Preventive Medicine and Public Health
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    • v.29 no.1 s.52
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    • pp.43-50
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    • 1996
  • We experienced a case of nephropathy in chronic lead poisoning. The patient was 43-year-old male who has been working in secondary lead smelting plant for 14 years. On admission, blood pressure was 160/90 mmHg and the others were non-specific. In past history, he received chelating agent administration for lead poisoning irregularly and medicated for gout, and the blood lead concentration was $180.0{\mu}g/dl$ on 2 months before admission. Smoking habit has been 1 pack per day for 15 years and drinking habit has been 1 bottle of Soju per day but less flow. In liver function test, AST/ALT were 27/28 IU/l and $\gamma-GT$ was 456 IU/l. In blood test, Hb : 11.5 g/dl, Hct : 34.0% and basophilic stipplings were found in peripheral blood smear. Chest PA was normal and abdominal ultrasonographic finding was non-specific except fatty liver. In the test of lead exposure indices, $PbB:83.0{\mu}g/dl,\;PbU:28.3{\mu}g/l$, and blood ZPP was $300.0{\mu}g/dl$. And in renal function test, BUN : 31.4 mg/dl, blood creatinine : 2.7mg/dl, blood uric acid. 9.1 mg/dl, urinary albumin : 100.0 mg/g creatinine, urinary $\alpha_1-microglobulin$ : 120.5 mg/g creatinine, urinary $\beta_2-microglobulin$ : $183.8{\mu}g/g$ creatinine, and 24 hours urinary creatinine clearance was 31.9 ml/min. The ultrasonoguided renal biopsy showed the global sclerosis of glomerulus, moderate atrophy and loss of tubule, and interstitial fibrosis in light microscopy. There were diffuse losses of brush border of proximal tubule in electronmicroscopy.

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