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Isolation and Identification of a Photosensitizer from Pueraria thunbergiana Leaves that Induces Apoptosis in SK-HEP-1 Cells (P. thunbergiana 잎으로부터 SK-HEP-1세포에 대한 apoptosis를 유도하는 광과민성물질의 분리 및 구조동정)

  • Lee, Jun Young;Kim, Mi Kyeong;Ha, Jun Young;Kim, Yong Gyun;Hong, Chang Oh;Kim, So Young;Kim, Chung-Hwan;Kim, Keun Ki
    • Journal of Life Science
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    • v.24 no.3
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    • pp.242-251
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    • 2014
  • The objective of this study was to isolate a photosensitizer from Pueraria thunbergiana leaves that induces apoptosis in SK-HEP-1 cells. Column chromatography and thin layer chromatography were used to isolate active compounds from extracts of P. thunbergiana leaves. The structures of the isolated compounds were determined by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. A substance, named M4-3, was purified from the leaves of P. thunbergiana using various chromatography methods, and the absorbance of the substance was measured. The absorbance was highest at 410 nm, suggesting that the M4-3 substance was a different compound from chlorophyll a and b, which absorb at 410, 502, 533, and 607 nm. Further analyses revealed that the M4-3 compound was a $13^2$-hydoxy pheophorbide, a methyl ester with a molecular weight of 662. M4-3 was identified as a derivative compound of pheophorbide, with a structure that magnesium comes away from the porphyrin ring. The results of the analysis of the cytotoxicity of the M4-3 substance against the SK-HEP-1 cells revealed that it inhibited rates of cell growth by 40% and 80% at a concentration of 0.04 ${\mu}M$ and 0.08 ${\mu}M$, respectively. The M4-3 compound was found to be a photosensitizer for cytotoxicity because it was appeared only in light condition as examining activity in different irradiation conditions (light condition and nonlight condition) under the same concentration. Analysis of morphological changes in the cells following cell death induced by exposure to the M4-3 substance reveled representative phenomena of apoptosis (nuclear condensation, vesicle formation, and fragmentation of DNA). The induction of apoptosis was attributed to the compound's photodynamic activity.

COMPARISON OF THE GROWTH RATES OF THREE STRAINS OF COMMON CARP, CYPRINUS CARPIO (잉어 3품종의 성장비교)

  • KIM In-Bae;JO Jae Yoon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.8 no.4
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    • pp.222-224
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    • 1975
  • Three strains of common carp, i. e. , Israeli carp, red-and-white, and golden strains, were stocked in the same pond, and their growth rates were compared with following results: From August 12 to November 21 in 1975, fingerlings of the three strains of common carp, Cyprinus carpo, each weighing about 0.5 g with total length of 2 to 3 cm, were stocked. The pond had an area of $316m^2$ with a mean water depth of 55cm, and the bottom was covered with a 20 to 30 cm thick layer of silt containing a considerable amount of decaying organic materials. Feed given was prepared with equal amounts of fish meal and polished barley, of which, in addition, $10\%$ green grass and $1\%$ table salt were mixed together when prepared into paste feed using a chopper after boiling the barley. Total protein content of the feed was $34.9\%$ in dry state with $5\%$ moisture content. Total feed given was 30.08 kg calculated in dry state to produce 20.588 kg of the common carp fingerlings, thus the feed coefficient being 1.51. By strains, the harvested Israeli carp ranged 98 to 311g each with a mean weight of $172.69g(100\%)$, red-and-white strain 15 to 318g with mean of $104.1g(60.3\%)$, and the golden strain 30 to 268g with mean of $128.7g(74.6\%)$. During the rearing season mean water temperature was $23.9^{\circ}C$ and the assumed main growth period with the water temperature above $15^{\circ}C$ was, upto the end of October, for 80 days with a mean water temperature of $23.9^{\circ}C$. Taking this main growth period as the basis for growth rate analysis, the mean daily increments, expressed as the attained body weight in times of the starting weight, become 1.075786 times (or the Israeli strain, 1.06901 times for the red-and-white strain, and 1.07185 times for the golden strain.

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Morphological Adaptation of Zostera marina L. to Ocean Currents in Korea (한국산 거머리말(Zostera marina L.)의 해류에 대한 형태적 적응)

  • Lim, Dong-Ok;Yun, Jang-Tak;Han, Kyung-Shik
    • Korean Journal of Environment and Ecology
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    • v.23 no.5
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    • pp.431-438
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    • 2009
  • The main purpose of this research is to prepare and provide basic materials for the propagational strategy of eelgrass by investigating on the morphological adaptation of Korean Zostera marina to ocean currents. An eelgrass plant mainly consists of rhizome, leaf sheath, leaves and roots. The rhizome is the horizontal stem of the plant that serves as the backbone from which the leaves and roots emerge. The leaf sheath is the bundle at the base of the leaves that holds the leaves together, protecting the meristem, the primary growth point of the shoot. Leaves originate from a meristem which is protected by a sheath at the actively growing end of the rhizome. As the shoot grows, the rhizome elongates, moving across or within the sediment, forming roots as it progresses. The aggregated leaves from the leaf sheath are found to have two cell layers on one side and multiple layers of airy tissues called aerenchyma on the other. The aerenchyma tissues are developed in multi-layered cell structures surrounding the veins which are formed in the leaf sheath. Generative shoots are made of rhizomes, which are circular or ovoidal, stem, and spathe and spadix. The transverse section of rhizome and the stem and central floral axis is found to be circular, ovoid and in the shape of convex respectively, and the vascular bundle, which is a part of transport system, has one large tube in the center and two small tubes on both sides. The layers of collenchyma cells numbered from 12 to 15 in the stem, and from 7 to 12 in the rhizome. The seed coat is composed of sclereids, small bundles of sclerenchyma tissues, which prevent the influx of sea water from the outside and help endure the environmental stress. In conclusion, alternative multi-layer structure in circular, convex type aggregated leaf base are interpreted to morphological adaption as doing tolerable elastic structure through movement of seawater. The generative shoots develop long slim stem and branches in circular or ovoidal shapes to minimize the adverse impacts of sea current, which can be interpreted as the plant's morphological adaptation to its environment.

Establishment of In Vitro 3-Dimensional Culture System of Mouse Endometrial Cells;I. Cytohistological Study on Mouse Endometrium (마우스 자궁내막 세포를 이용한 3차원적 배양시스템 확립에 관한 연구; I. 마우스 자궁내막에 관한 세포조직학적 연구)

  • Nam, Hwa-Kyung;Kim, Eun-Young;Lee, Keum-Sil;Park, Sae-Young;Park, Eun-Mi;Kwon, Jung-Kyun;Yoon, San-Hyun;Park, Se-Pil;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.1
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    • pp.31-37
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    • 2000
  • This study was designed to identify the ultrastructural changes of mouse endometrium during peri-implantation period and obtain the fundamental information for the establishment of 3-dimensional culture system of mouse endometrial cells in vitro. The used female ICR mice ($6{\sim}8$ wks) were conducted on pregnant. The biopsies were obtained from whole uterus at cycle day 1 (D1) and day 5 (D5) after hCG injection and mating. The biopsies materials were fixed 2.5% glutaraldehyde and 1% osmium tetroxide. Subsequently, for observation using light and transmission electron microscopy (LM and TEM), they were dehydrated and embedded in Epon and the embedded biopsies were sectioned and stained. For scanning electron microscopy (SEM), the fixed specimens were dehydrated, dried and coated with gold. 1) For LM, the biopsied materials at D5 (late secretory phase) were appeared the extended stromal layer by increased connective tissues and the fully developed endometrial glands and vessels compared with D1 (early secretory phase). 2) For TEM, the mouse endometrium was consisted of 3-layers, a simple polarized columnar epithelial cells, basement membrane and stromal cells. At D5, the distribution of microvilli, endoplasmic reticulum, Golgi body, lipid and glycogen deposits, secretory granules and surface area of basement membrane were increased. 3) For SEM, the degree of folding and microvilli of surface of mouse epithelial cells was became more and more according to the process of secretory phase, and at D5, implantation time of mouse, the appearance of pinopodes as a specific marker of uterine receptivity was found. The uterine pinopodes of mouse were found in narrow sites at the luminal surface, irregularity and appeared the different stages in the same sample. Therefore, these results indicated that the mouse endometrium was experienced dramatic morphological changes during peri-implantation period.

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Annealing Effects on Properties of ZnO Nanorods Grown by Hydrothermal Method (수열합성법으로 성장된 산화아연 나노막대의 특성 및 열처리 효과)

  • Jeon, Su-Min;Kim, Min-Su;Kim, Ghun-Sik;Cho, Min-Young;Choi, Hyun-Young;Yim, Kwang-Gug;Kim, Hyeoung-Geun;Lee, Dong-Yul;Kim, Jin-Soo;Kim, Jong-Su;Lee, Joo-In;Leem, Jae-Young
    • Journal of the Korean Vacuum Society
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    • v.19 no.4
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    • pp.293-299
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    • 2010
  • Vertically aligned ZnO nanorods on Si (111) substrate were prepared by hydrothermal method. The ZnO nanorods on spin-coated seed layer were synthesized at $140^{\circ}C$ for 6 hours in autoclave and were thermally annealed in argon atmosphere for 20 minutes at temperature of 300, 500, $700^{\circ}C$. The effects of the thermal annealing on the structural and optical properties of the grown on ZnO nanorods were investigated by X-ray diffraction (XRD), field-emission scanning electron microscopy (FE-SEM), photoluminescence (PL). All the ZnO nanorods show a strong ZnO (002) and weak (004) diffraction peak, indicating c-axis preferred orientation. The residual stress of the ZnO nanorods is changed from compressive to tensile by increasing annealing temperature. The hexagonal shaped ZnO nanorods are observed. The PL spectra of the ZnO nanorods show a sharp near-band-edge emission (NBE) at 3.2 eV, which is generated by the free-exciton recombination and a broad deep-level emission (DLE) at about 2.12~1.96 eV, which is caused by the defects in the ZnO nanorods. The intensity of the NBE peak is decreased and the DLE peak is red-shifted due to oxygen-related defects by thermal annealing.

Caffeine treatment during in vitro maturation improves developmental competence of morphologically poor oocytes after somatic cell nuclear transfer in pigs (돼지 난자의 체외성숙에서 Caffeine 처리가 난자 성숙과 체세포 핵이식 배아의 체외발육에 미치는 영향)

  • Lee, Joohyeong;You, Jinyoung;Lee, Hanna;Shin, Hyeji;Lee, Geun-Shik;Lee, Seung Tae;Lee, Eunsong
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.131-138
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    • 2017
  • In most mammals, metaphase II (MII) oocytes having high maturation promoting factor (MPF) activity have been considered as good oocytes and then used for assisted reproductive technologies including somatic cell nuclear transfer (SCNT). Caffeine increases MPF activity in mammalian oocytes by inhibiting p34cdc2 phosphorylation. The objective of this study was to investigate the effects of caffeine treatment during in vitro maturation (IVM) on oocyte maturation and embryonic development after SCNT in pigs. To this end, morphologically good (MGCOCs) and poor oocytes (MPCOCs) based on the thickness of cumulus cell layer were untreated or treated with 2.5 mM caffeine during 22-42, 34-42, or 38-42 h of IVM according to the experimental design. Caffeine treatment for 20 h during 22-42 h of IVM significantly inhibited nuclear maturation compared to no treatment. Blastocyst formation of SCNT embryos was not influenced by the caffeine treatment during 38-42 h of IVM in MGCOCs (41.1-42.1%) but was significantly improved in MPCOCs compared to no treatment (43.4 vs. 30.1%, P<0.05). No significant effects of caffeine treatment was observed in embryo cleavage (78.7-88.0%) and mean cell number in blastocyst (38.7-43.5 cells). The MPF activity of MII oocytes in terms of p34cdc2 kinase activity was not influenced by the caffeine treatment in MGCOCs (160.4 vs. 194.3 pg/ml) but significantly increased in MPCOCs (133.9 vs. 204.8 pg/ml). Our results demonstrate that caffeine treatment during 38-42 h of IVM improves developmental competence of SCNT embryos derived from MPCOCs by influencing cytoplasmic maturation including increased MPF activity in IVM oocytes in pigs.

Establishment of the Appropriate Risk Standard through the Risk Assessment of Accident Scenario (사고시나리오별 위험도 산정을 통한 적정 위험도 기준 설정)

  • Kim, Kun-Ho;Chun, Young-Woo;Hwang, Yong-Woo;Lee, Ik-Mo;Kwak, In-ho
    • Journal of Korean Society of Environmental Engineers
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    • v.39 no.2
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    • pp.74-81
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    • 2017
  • An off-site consequence analysis is used to calculate the risks when hazardous chemicals that is being used on-site has been exposed off-site; the biggest factor that impacts the risk is the risks of accident scenarios. This study seeks to calculate risks according to accident scenarios by applying OGP/LOPA risk calculating methods for similar facilities, calculate risk reduction ratio by inspecting applicable IPL for incidents, and propose an appropriate risk standard for different risk calculating methods. Considering all applicable IPL when estimating the safety improvement of accident scenarios, the risk of OGP is 8.05E-04 and the risk of LOPA is 1.00E-04, According to the case of IPL, the risk is 1.34E-02. The optimal risk level for accident scenarios using LOPA was $10^{-2}$, but the appropriate risk criteria for accident scenarios in foreign similar studies were $10^{-3}{\sim}10^{-4}$, the risk of a scenario can be determined at an unacceptable level. When OGP is applied, it is analyzed as acceptable level, but in case of applying LOPA, all applicable IPL should be applied in order to satisfy the acceptable risk level. Compared to OGP, the risk is high when LOPA is applied. Therefore, the acceptable risk level should be set differently for each risk method.

MICROLEAKAGE OF COMPOSITE RESIN RESTORATION ACCORDING TO THE NUMBER OF THERMOCYCLING (열순환 횟수에 따른 복합레진의 미세누출)

  • Kim, Chang-Youn;Shin, Dong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.32 no.4
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    • pp.377-384
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    • 2007
  • Present tooth bonding system can be categorized into total etching bonding system (TE) and self-etching boding system (SE) based on their way of smear layer treatment. The purposes of this study were to compare the effectiveness between these two systems and to evaluate the effect of number of themocycling on microleakage of class V composite resin restorations. Total forty class V cavities were prepared on the single-rooted bovine teeth and were randomly divided into four experimental groups: two kinds of bonding system and another two kinds of thermocycling groups. Half of the cavities were filed with Z250 following the use of TE system, Single Bond and another twenty cavities were filled with Metafil and AQ Bond, SE system. All composite restoratives were cured using light curing unit (XL2500, 3M ESPE, St. Paul, MN, USA) for 40 seconds with a light intensity of $600mW/cm^2$. Teeth were stored in distilled water for one day at room temperature and were finished and polished with Sof-Lex system. Half of teeth were thermocycled 500 times and the other half were thermocycled 5,000 times between $5^{\circ}C$ and $55^{\circ}C$ for 30 second at each temperature. Teeth were isolated with two layers of nail varnish except the restoration surface and 1 mm surrounding margins. Electrical conductivity (${\mu}A$) was recorded in distilled water by electrochemical method. Microleakage scores were compared and analyzed using two-way ANOVA at 95% level. From this study, following results were obtained: There was no interaction between variables of bonding system and number of thermocycling (p = 0.485). Microleakage was not affected by the number of thermocycling either (p = 0.814). However, Composite restoration of Metafil and AQ Bond, SE bond system showed less microleakage than composite restoration of Z250 and Single Bond, TE bond system (p = 0.005).

Mixed Osteosarcoma with Metastatic Alveolar Carcinomatous Appearance in Canine Mammary Gland Tumor (개의 유선에서 발생한 혼합성 골육종 및 샘포 상피세포의 전이성 암종)

  • Goo, Moon-Jung;Hong, Il-Hwa;Park, Jin-Kyu;Yang, Hai-Jie;Yuan, Dong-Wei;Ki, Mi-Ran;Lee, Hye-Rim;Hong, Kyung-Sook;Han, Jung-Youn;Hwang, Ok-Kyung;Kim, Tae-Hwan;Do, Sun-Hee;Jeong, Kyu-Shik
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1766-1770
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    • 2007
  • We describe here a case of malignant mixed osteogenic tumor of the mammary gland with alveolar carcinomatous appreance. A firm, 2 to 2.5cm (in diameter) mass under the 5th nipple, showing the structure of extraosseous osteogenic sarcoma, was removed from the left 5th mammary gland of 12-year-old female dog. When investigated under the microscope, the osteoid material undergoing mineralization was surrounded by numerous scattered osteoblasts and a few osteoclastic cells throughout the osteoid tumorous stroma. The osteoid lesions were continuous with hypercellular myoepithelial cells of a very immature character with several mitotic figures. In addition, there were also carcinomatous tubules and alveoli, with invading cells into peripheral stroma, surrounded by myoepithelial cells in the mammary gland. In these lesions, emanating cords of tumor cells appear to be continuous with the myoepithelial cell layer of a duct. The presence of all these cell types suggests the existence of a common malignant origin, the stem cell being differentiated into epithelial carcinomatous and mesenchymal sarcomatous chondral and osteogenic tissues.

Effects of acid-treatment conditions on the surface properties of the RBM treated titanium implants (산-처리 조건이 RBM처리한 티타늄 임플란트의 표면 특성에 주는 영향)

  • Lee, Han-Ah;Seok, Soohwang;Lee, Sang-Hyeok;Lim, Bum-Soon
    • Korean Journal of Dental Materials
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    • v.45 no.4
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    • pp.257-274
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    • 2018
  • The purpose of this study was to evaluate the effect of acid-treatment conditions on the surface properties of the RBM (Resorbable Blast Media) treated titanium. Disk typed cp-titanium specimens were prepared and RBM treatments was performed with calcium phosphate ceramic powder. Acid solution was mixed using HCl, $H_2SO_4$ and deionized water with 4 different volume fraction. The RBM treated titanium was acid treated with different acid solutions at 3 different temperatures and for 3 different periods. After acid-treatments, samples were cleaned with 1 % Solujet solution for 30 min and deionized water for 30 min using ultrasonic cleanser, then dried in the electrical oven ($37^{\circ}C$). Weight of samples before and after acid-treatment were measured using electric balance. Surface roughness was estimated using a confocal laser scanning microscopy, crystal phase in the surface of sample was analyzed using X-ray diffractometer. Surface morphology and components were evaluated using Scanning Electron Microscope (SEM) with Energy Dispersive X-ray spectroscopy (EDX) and X-ray Photoemission Spectroscopy (XPS). Values of the weight changes and surface roughness were statistically analyzed using Tukey-multiple comparison test (p=0.05). Weight change after acid treatments were significantly increased with increasing the concentration of $H_2SO_4$ and temperature of acid-solution. Acid-treatment conditions (concentration of $H_2SO_4$, temperature and time) did not produce consistent effects on the surface roughness, it showed the scattered results. From XRD analysis, formation of titanium hydrides in the titanium surface were observed in all specimens treated with acid-solutions. From XPS analysis, thin titanium oxide layer in the acid-treated specimens could be evaluated. Acid solution with $90^{\circ}C$ showed the strong effect on the titanium surface, it should be treated with caution to avoid the over-etching process.