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Siamese Crocodile White Blood Cell Extract Inhibits Cell Proliferation and Promotes Autophagy in Multiple Cancer Cell Lines

  • Phosri, Santi;Jangpromma, Nisachon;Chang, Leng Chee;Tan, Ghee T.;Wongwiwatthananukit, Supakit;Maijaroen, Surachai;Anwised, Preeyanan;Payoungkiattikun, Wisarut;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.1007-1021
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    • 2018
  • Cancer represents one of the most significant threats to human health on a global scale. Hence, the development of effective cancer prevention strategies, as well as the discovery of novel therapeutic agents against cancer, is urgently required. In light of this challenge, this research aimed to evaluate the effects of several potent bioactive peptides and proteins contained in crocodile white blood cell extract (cWBC) against LU-1, LNCaP, PC-3, MCF-7, and CaCo-2 cancer cell lines. The results demonstrate that 25, 50, 100, and $200{\mu}g/ml$ cWBC exhibits a strong cytotoxic effect against all investigated cell lines ($IC_{50}$ $70.34-101.0{\mu}g/ml$), while showing no signs of cytotoxicity towards noncancerous Vero and HaCaT cells. Specifically, cWBC treatment caused a significant reduction in the cancerous cells' colony forming ability. A remarkable suppression of cancerous cell migration was observed after treatment with cWBC, indicating potent antimetastatic properties. The mechanism involved in the cancer cell cytotoxicity of cWBC may be related to apoptosis induction, as evidenced by typical apoptotic morphology features. Moreover, certain cWBC concentrations induced significant overproduction of ROS and significantly inhibited the $S-G_2/M$ transition in the cancer cell. The molecular mechanisms of cWBC in apoptosis induction were to decrease Bcl-2 and XIAP expression levels and increase the expression levels of caspase-3, caspase-8, and p53. These led to a decrease in the expression level of the cell cycle-associated gene cyclin-B1 and the arrest of cell population growth. Consequently, these findings demonstrate the prospect of the use of cWBC for cancer therapy.

Enhanced Mechanical Properties and Thermal Stability of CrAlN Coatings by Interlayer Deposition (중간층 증착에 의한 CrAlN 코팅의 기계적 물성 및 내열성 향상에 관한 연구)

  • Kim, Hoe-Geun;Ra, Jeong-Hyeon;Song, Myeon-Gyu;Lee, Sang-Yul
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2016.11a
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    • pp.100-100
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    • 2016
  • 물리기상증착방법 (Physical vapor deposition)에 의하여 합성된 천이금속 질화물 박막은 경도, 내마모성 등 절삭공구의 성능을 향상시키며, Ti-Al-N, Ti-Zr-N, Zr-Al-N, Cr-Si-N 등의 3원계 경질 박막에 대한 연구가 지속적으로 이루어지고 있다. 이중에서도 CrAlN 코팅은 높은 경도, 낮은 표면 조도 등의 우수한 기계적 특성 이외에 고온에서 안정한 합금상 형성으로 인하여 우수한 내열성을 보유하여 공구 코팅으로의 적용 가능성이 크다. 그러나 최근 공구사용 환경의 가혹화로 인하여 코팅의 내마모성 및 내열성 등의 물성 향상을 통한 공구의 수명 향상이 필요시 되고 있으며, 코팅에 적합한 중간층을 합성함으로써 공구 코팅으로의 적용 가능성을 높이는 연구들이 진행되고 있다. 본 연구에서는 CrAlN 코팅의 물성을 향상시키기 위해 CrAlN 코팅과 WC-Co 6wt.% 모재 사이에 CrN, CrZrN, CrN/CrZrSiN 등의 다양한 중간층을 합성하였으며, 중간층을 포함한 모든 코팅의 두께는 $3{\mu}m$ 로 제어하였다. 합성된 코팅의 미세조직, 경도 및 탄성계수, 내모성을 분석하기 위해 field emission scanning electron microscopy(FE-SEM), nano-indentation, ball-on-disk 마모시험기 및 ${\alpha}-step$을 사용하였다. 코팅의 내열성을 확인하기 위해 코팅을 furnace에 넣어 공기중에서 500, 600, 700, 800, 900, $1,000^{\circ}C$로 30분 동안 annealing 한 후에 nano-indentation을 사용하여 경도를 측정하였다. CrAlN 코팅을 나노 인덴테이션으로 분석한 결과, 모든 코팅의 경도(35.5-36.2 GPa)와 탄성계수(424.3-429.2 GPa)는 중간층의 종류에 상관없이 비슷한 값을 보인 것으로 확인됐다. 그러나, 코팅의 마찰계수는 중간층의 종류에 따라 다른 값을 보였으며, CrN/CrZrSiN 중간층을 증착한 CrAlN 코팅의 마찰계수는 0.34로 CrZrN 중간층을 증착한 CrAlN 코팅의 마찰계수(0.41)에 비해 낮은 값을 보였다. 또한, 코팅의 마모율 및 마모폭도 비슷한 경향을 보인 것으로 보아, CrN/CrZrSiN 중간층을 합성한 CrAlN 코팅의 내마모성이 상대적으로 우수한 것으로 판단된다. 이것은 중간층의 H/E ratio가 코팅의 내마모성에 미치는 영향에 의한 결과로 사료된다. H/E ratio는 파단시의 최대 탄성 변형율로써, 모재/중간층/코팅의 H/E ratio 구배에 따라 코팅 내의 응력의 완화 정도가 변하게 된다. WC 모재 (H/E=0.040)와 CrAlN 코팅(H/E=0.089) 사이에서 CrN, CrZrSiN 중간층의 H/E ratio 는 각각 0.076, 0.083 으로 모재/중간층/코팅의 H/E ratio 구배가 점차 증가함을 확인 할 수 있었고, 일정 응력이 지속적으로 가해지면서 진행되는 마모시험중에 CrN과 CrZrSiN 중간층이 WC와 CrAlN 코팅 사이에서 코팅 내부의 응력구배를 완화시키는 역할을 함으로써 CrAlN 코팅의 내마모성이 향상된 것으로 판단된다. 모든 코팅을 열처리 후 경도 분석결과, CrN/CrZrSiN 중간층을 증착한 CrAlN 코팅은 $1,000^{\circ}C$까지 약 28GPa의 높은 경도를 유지한 것으로 확인 되었고, 이는 CrZrSiN 중간층 내에 존재하는 $SiN_x$ 비정질상의 우수한 내산화성에 의한 결과로 판단된다.

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Properties of TiN films prepared by using the DC sputtering and HIPIMS. (DC 스퍼터링과 HIPIMS로 제조한 TiN 박막의 특성 비교)

  • Byeon, In-Seop;Yang, Ji-Hun;Jeong, Jae-Hun;Kim, Seong-Hwan;Jeong, Jae-In
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2016.11a
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    • pp.102-102
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    • 2016
  • 본 연구에서는 직류 전원(direct current; DC)을 이용한 스퍼터링과 고전력펄스 마그네트론 스퍼터링(high-power impulse magentron sputtering; HIPIMS)의 두 가지 방법과 빗각 증착을 적용하여 제조한 티타늄 질화물(TiN) 박막의 미세구조 변화가 물성에 미치는 영향을 확인하였다. TiN 박막은 99.5%의 Ti 타겟을 사용하고, Ar가스와 $N_2$ 분위기에서 스테인리스(SUS304)와 초경(cdmented carbide; WC-10wt.%Co) 기판위에 코팅하였다. 기판은 알코올과 아세톤으로 초음파 세척을 실시한 후 진공용기에 장착하고 기본 진공도인 ${\sim}2.0{\times}10^{-5}Torr$ 까지 진공배기를 실시하였다. 기판과 타겟 간의 거리는 DC 스퍼터링은 10 cm, HIPIMS 스퍼터링은 8.5 cm 이었다. 진공용기의 압력이 기본 진공도까지 배기되면 Ar 가스를 ${\sim}10^{-2}Torr$로 주입한 후 기판에 라디오 주파수(radio frequency; RF) 전원으로 약 -800 V의 전압을 인가하여 글로우 방전을 발생시키고 약 30 분간 청정을 실시하였다. 기판의 청정이 끝난 후 기본 진공도까지 배기한 후 Ar와 $N_2$ 가스를 ${\sim}10^{-3}Torr$로 주입하여 TiN 코팅을 실시하였다. 빗각의 크기는 $45^{\circ}$$-45^{\circ}$이며, TiN 박막의 총 두께는 약 $2.5{\sim}4.0{\mu}m$ 로 유지하였다. 공정조건에 따라 TiN 박막의 주상정은 형태와 기울어진 각도가 다른 것을 확인하였다. DC 스퍼터링으로 제조된 TiN 박막은 기판홀더에 약 -100 V 의 bias 전압을 인가하면 인가하지 않은 박막에 비해 치밀한 박막의 성장과 경도 값도 증가하는 사실을 확인하였다. 또한 빗각을 적용하고 bias 전압을 인가하지 않은 시편에서 박리현상이 일어났다. HIPIMS로 제조한 TiN 박막은 bias 전압을 인가한 박막과 인가하지 않은 박막의 주상정 형상과 경도 값에 큰 차이가 없었으며, 박막의 박리현상은 모든 시편에서 일어나지 않았다. DC 스퍼터링으로 제조한 TiN 박막은 bias 전압을 인가하지 않으면 색상이 노란색이 아닌 갈색으로 나타났으며, HIPIMS으로 제조한 박막은 bias 전압 인가 유무에 상관없이 노란색 색상을 나타냈다. 앞서 설명한 DC 스퍼터링과 HIPIMS의 공정조건에 따라 나타난 박막의 경도, 색상, 물성변화 차이는 DC 스퍼터링보다 높은 HIPIMS의 이온화율에서 기인한 것으로 생각된다. 본 연구결과를 이용하면 다양한 형태의 박막 구조 제어가 가능하고 이러한 미세구조 제어를 통해서 박막의 물성도 제어가 가능할 것으로 판단된다.

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Dry Enteric Coating Process of Lactic Acid Bacteria by Hybridization System (Hybridization system을 이용한 유산균의 장용성 건식 피복)

  • Park, Dong-June;An, Eun-Young;Kim, Jae-Seung;Imm, Jee-Young;Han, Kyoung-Sik;Kim, Sae-Hun;Oh, Se-Jong
    • Korean Journal of Food Science and Technology
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    • v.34 no.5
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    • pp.856-861
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    • 2002
  • Surface-modified powders were produced by hybridization system using core freeze-dried lactic acid bacteria (Lactobacillus acidophilus ATCC 43121) and enteric coating materials. Scanning electron microscopy showed that the surface of freeze-dried lactic acid bacteria changed to smooth round shape during surface reforming process, although no significant physical damages affecting the activity of the lactic acid bacteria were observed based on viability and salt-tolerance tests. Signigicant difference was not found in acid tolerance test probably due to the inherent acid tolerance of L. acidophilus ATCC 43121. Significantly improved heat tolerance was obtained by surface modification process. Among the tested coating materials, Sureteric showed a higher surface- reforming ability than Eudragit S100 and L100-55. Core : coating ratio agent of 9 : 1 (w/w) with rotor speed of 15,000 rpm for 3 min were determined to be optimum conditions for the process.

Protection of Infection and Eradication Activity of Culture Product by Pediococcus pentosaceus CBT SL4 Showing Antimicrobial Activity against Helicobacter pylori (Helicobacter pylori에 대한 항균활성을 나타내는 Pediococcus pentosaceus CBT SL4 배양물의 감염방어 및 제균활성)

  • Hong, Un-Pyo;Chung, Myung-June;Kim, Soo-Dong;Oh, Eun-Taex;So, Jae-Seong;Chung, Chung-Il
    • Korean Journal of Food Science and Technology
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    • v.36 no.5
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    • pp.779-783
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    • 2004
  • New food ingredient was developed to eradicate and protect against re-infection of Helicobacter pylori in fermentation broth of lactic acid bacteria (LAB) showing antimicrobial activity against pathogenic microorganisms such as H. pylori and Listeria monocytogenes. LAB strain CBT SL4 was identified as Pediococcus pentosaceus by 16S rDNA sequencing and its culture broth showed antimicrobial activity of 800 AU/mL against H. pylori in optimized fermentation process. Using thin layer concentration system and spray-typed fluid bed drier system, concentrated powder product showing activity of 12,800 AU/g was harvested. Product showed eradication and protection activities against H. pylori infection on feeding test (50 AU/day) using Mongolian gerbil infection model. After 4 weeks therapy of 8,000 AU/day, ${\Delta}13CO_2$ level (DOB30) decreased about 40% in urea breath test on patient with H. pylori infection. Result show concentrated culture product of P. pentosaceus CBT SL4 has eradicating and protecting activities against H. pylori infection and can be used as food-active ingredient for prevention of gastric and duodenum ulcer caused by H. pylori.

Screening and Extraction Condition of Antiaging Bioactive Substances from Medicinal Plants (각종 약용 식물로부터 노화 억제 관련 생리활성 물질의 탐색 및 추출 조건)

  • Yu, Hyung-Eun;Dela Paz, Leaniza Michella M.;Bae, Young-Joo;Lee, Dae-Hyoung;Park, Jong-Sang;Kwak, Hahn-Shik;Kim, Ha-Kun;Lee, Jong-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1136-1142
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    • 2005
  • To develop new anti-aging foods or cosmetics by using antioxidants, SOD activator and elastase inhibitor, both potent anti-aging substances, were screened from various extracts of medicinal Plants and its optimal extraction conditions were investigated. Antioxidant activity has showed the highest in methanol extracts of Prunus persica (seed; 98.0$\%$). Methanol extracts of Morus alba (leave; 41.0$\%$) showed the highest elastase inhibitory activity while Lycium chinense (fruit; 197$\%$) showed the highest activation effect in SOD activity. The Prunus persica extract that exhibited the highest activity was extracted by treatment of Prunus persica powder with methanol at 40$^{\circ}C$ for 18 h and the SOD activity was maximum with extract from Lycium chinense extracted with deionized water at 30$^{\circ}C$ for 12 h. Elastase inhibitory activity of Morus alba was maximally extracted when it was treated with 70$\%$ methanol at 50$^{\circ}C$ for 12 h.

Effect of Lentinus edodes and Pleurotus eryngii Extracts on Proliferation and Apoptosis in Human Colon Cancer Cell Lines (표고와 새송이버섯이 대장암 세포 증식 및 세포사멸에 미치는 영향)

  • 황용주;남혜경;장문정;노건웅;김선희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.2
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    • pp.217-222
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    • 2003
  • We studied effects of hot water extract of Lentinus edodes (Berk.)sing. and Pleurotus eryngii (De Candolle ex Fries) Quel mushroom on proliferation and apoptosis of the human colon adenocarcinoma, HT-29 and Caco-2.. Cells were maintained with Dulbecco's modified Eagle medium/Ham's F-12 nutrient mixture supplemented with 10% fetal bovine serum at 37$^{\circ}C$ in a humidified $CO_2$. For cell proliferation experiments, cells were seeded in 35 mm dishes, treated with the various concentrations of the extract for the different time course. Apoptosis was measured by caspase-3 activity The more contents of the extract added in HT-29 and Caco-2 were, the more cell proliferation was suppressed. When we incubated HT-29 cells for 24, B\ulcorner72, and 96 hours after treatments, cell proliferation was markedly suppressed after 96 hours. Also, caspase-3 activity in HT-29 was increased by the treatment of Lentinus edodes and Pleurotus eryngii extracts. However, the treatment of the extract to SNU484, Korean stomach adenocarcinoma, did not show any influence on cell proliferation and caspase-3 activity Therefore, Lentinus edodes and Pleurotus eryngii are strongly recommended for the prevention and treatment of colon cancer.

Vehicle Detection using Feature Points with Directional Features (방향성 특징을 가지는 특징 점에 의한 차량 검출)

  • Choi Dong-Hyuk;Kim Byoung-Soo
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.42 no.2 s.302
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    • pp.11-18
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    • 2005
  • To detect vehicles in image, first the image is transformed with the steerable pyramid which has independent directions and levels. Feature vectors are the collection of filter responses at different scales of a steerable image pyramid. For the detection of vehicles in image, feature vectors in feature points of the vehicle image is used. First the feature points are selected with the grid points in vehicle image that are evenly spaced, and second, the feature points are comer points which m selected by human, and last the feature points are corner Points which are selected in grid points. Next the feature vectors of the model vehicle image we compared the patch of the test images, and if the distance of the model and the patch of the test images is lower than the predefined threshold, the input patch is decided to a vehicle. In experiment, the total 11,191 vehicle images are captured at day(10,576) and night(624) in the two local roads. And the $92.0\%$ at day and $87.3\%$ at night detection rate is achieved.

Preparation of LiFe PO4 Using Chitosan and its Cathodic Properties for Rechargeable Li-ion Batteries

  • Hong, Kyong-Soo;Yu, Seong-Mi;Ha, Myoung-Gyu;Ahn, Chang-Won;Hong, Tae-Eun;Jin, Jong-Sung;Kim, Hyun-Gyu;Jeong, Euh-Duck;Kim, Yang-Soo;Kim, Hae-Jin;Doh, Chil-Hoon;Yang, Ho-Soon;Jung, Hee
    • Bulletin of the Korean Chemical Society
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    • v.30 no.8
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    • pp.1719-1723
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    • 2009
  • The LiFeP$O_4$ powder was synthesized by using the solid state reaction method with Fe($C_2O_4){\cdot}2H_2O,\;(NH_4)_2HPO_4,\;Li_2CO_3$, and chitosan as a carbon precursor material for a cathode of a lithium-ion battery. The chitosan added LiFePO4 powder was calcined at 350 ${^{\circ}C}$ for 5 hours and then 800 ${^{\circ}C}$ for 12 hours for the calcination. Then we calcined again at 800 ${^{\circ}C}$ for 12 hours. We characterized the synthesized compounds via the crystallinity, the valence states of iron ions, and their shapes using TGA, XRD, SEM, TEM, and XPS. We found that the synthesized powders were carbon-coated using TEM images and the iron ion is substituted from 3+ to 2+ through XPS measurements. We observed voltage characteristics and initial charge-discharge characteristics according to the C rate in LiFeP$O_4$ batteries. The obtained initial specific capacity of the chitosan added LiFeP$O_4$ powder is 110 mAh/g, which is much larger than that of LiFeP$O_4$ only powder.

Studies on the Anti-inflammatory and Anti-apoptotic Effect of Catalposide Isolated from Catalpa ovata (개오동나무에서 추출(抽出)한 Catalposide의 항염(抗炎) 및 세포고사(細胞枯死) 억제효과(抑制效果)에 관(關)한 연구(硏究))

  • Oh, Cheon-Sik;Hwang, Sang-Wook;Kim, Yong-Woo;Song, Dal-Soo;Chae, Young-Seok;Jeong, Jong-Gil;Song, Ho-Joon;Shin, Min-Kyo
    • The Korea Journal of Herbology
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    • v.20 no.3
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    • pp.29-41
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    • 2005
  • Objectives : The use of natural products with therapeutic properties is as ancient as human civilisation and, for a long time, mineral, plant and animal products were the main sources of drugs. Catalposide, the major iridoid glycoside isolated from the stem bark of Catalpa ovata G. Don (Bignoniceae) has been shown to possess anti-microbial and anti-tumoral properties. Heme oxygenase-1 (HO-1) is a stress response protein and is known to play a protective role against the oxidative injury. In this study, we examined whether catalposide could protect Neuro 2A cells, a kind of neuronal cell lines, from oxidative damage through the induction of HO-1 protein expression and HO activity. We also examined the effects of catalposide on the productions of tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ and nitric oxide (NO) on RAW 264.7 macrophages activated with the endotoxin lipopolysaccharide. Methods : HO-1 expression in Neuro 2A cells was measured by Western blotting analysis. NO and $TNF--{\alpha}$ produced by RAW 264.7 macrophage were measured by Griess reagent and enzyme-linked immunosorbent assay, respectively. Results : The treatment of the cells with catalposide resulted in dose- and time-dependent up-regulations of both HO-1 protein expression and HO activity. Catalposide protected the cells from hydrogen peroxide-induced cell death. The protective effect of catalposide on hydrogen peroxide-induced cell death was abrogated by zinc protoporphyrin IX, a HO inhibitor. Additional experiments revealed the involvement of CO in the cytoprotective effect of catalposide-induced HO-1. In addition, catalposide inhibited the productions of $TNF--{\alpha}$ and NO with significant decreases in mRNA levels of $TNF--{\alpha}$ and inducible NO synthase. Conclusions : Our results indicate that catalposide is a potent inducer of HO-1 and HO-1 induction is responsible for the catalposide-mediated cytoprotection against oxidative damage and that catalposide may have therapeutic potential in the control of inflammatory disorders.

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