• Title/Summary/Keyword: $Mg^{2+}$

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Isolation and Structure Identification of Photosensitizer from Perilla frutescens Leaves Which Induces Apoptosis in U937 (들깻잎(Perilla frutescens)으로부터 U937 세포에 apoptosis를 유도하는 광과민성 물질의 분리 및 구조동정)

  • Ha, Jun Young;Kim, Mi Kyeong;Lee, Jun Young;Choi, Eun Bi;Hong, Chang Oh;Lee, Byong Won;Bae, Chang Hwan;Kim, Keun Ki
    • Journal of Life Science
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    • v.25 no.1
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    • pp.53-61
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    • 2015
  • In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the photosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an ethyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magnesium comes away from a porphyrin ring. Observation of morphological changes in U937 cells following cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cytotoxicity of PL9443 substance against U937 cells showed that inhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.

Antioxidant Activities of Stachys sieboldii MIQ Roots (초석잠(Stachys sieboldii MIQ) 뿌리 추출물의 항산화 활성 고찰)

  • 백홍석;나영수;김도한;이창한;류병호;송승구
    • Journal of Life Science
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    • v.14 no.1
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    • pp.1-7
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    • 2004
  • The root of Stnchys Sieboldif MIQ was extracted three times with methanol and extract was found to contain 3.02% of polyphenols and 1.97% of flavonoids. DPPH radical scavenging method, ferric thiocyanate method, and nitrite scavenging ability method were employed to investigate the constituents of the extract and to measure their activity on antioxidation. The fraction extracted by ethylacetate showed higher anti oxidation value than that of $\alpha-tocopherol$, butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) at the same concentration. UV-VIS spectral data of the extract by ethylacetate that was isolated on a silica gel column proved adsorption maxima in the range of 280∼330 nm. The fraction ES-RS that has $\lambda_{max}(nm)$ of band 1, 325nm and band II, 289nm exhibitd the strongest activity on antioxidation. ES-R5 fraction showed similar pattern to flavones by the analysis of UV-VIS spectral data.

Effect of Slurry Composting Bio-filtration (SCB) by Subsurface Drip Fertigation on Cucumber (Cucumis sativus L.) Yield and Soil Nitrogen Distribution in Greenhouse

  • Lim, Tae-Jun;Park, Jin-Myeon;Noh, Jae-Seung;Lee, Seong-Eun;Kim, Ki-In
    • Korean Journal of Soil Science and Fertilizer
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    • v.46 no.4
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    • pp.253-259
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    • 2013
  • The use of subsurface drip fertigation using slurry composting bio-filtration (SCB) as nitrogen (N) fertilizer source can be beneficial to improve fertilizer management decision. The objective of this study was to evaluate effects of SCB liquid fertilizer by subsurface drip fertigation on cucumber (Cucumis sativus L.) yield and soil nitrogen (N) distribution under greenhouse condition. Cucumber in greenhouse was transplanted on April $4^{th}$ and Aug $31^{st}$ in 2012. N sources were SCB and urea. Four N treatments with 3 replications consisted of control (No N fertilizer), SCB 0.5N + Urea 0.5N (50:50 split application), SCB 1.0N, Urea 1.0N. 100% of N recommendation rate from soil testing was denoted as 1.0N. The subsurface drip line and a tensiometer were installed at 30 cm soil depth. An irrigation was automatically started when the tensiometer reading was -15 kPa. The growth of cucumber at 85 days after transplanting was 5% higher in all N treatment than control. Semi-forcing culture produced more fruit yield than retarding culture. Fruit yields were 62.2, 76.3, 76.4, and 75.1 Mg $ha^{-1}$ for control, SCB 1.0N, Urea 1.0N, and SCB 0.5N + Urea 0.5N, respectively. Although fruit yields were similar under SCB 1.0N, Urea 1.0N, and SCB 0.5N + Urea 0.5N, 176 kg K $ha^{-1}$ can be over applied if cucumber is grown twice a year under SCB 1.0N that may result in K accumulation in soil. N uptake was 172, 209, 213, 207 kg $ha^{-1}$ for control, SCB 1.0N, Urea 1.0N, and SCB 0.5N + Urea 0.5N, respectively. N use efficiency was the highest (37%) at SCB 0.5N + Urea 0.5N under semi-forcing culture. Nitrate-N concentration in soil for all N treatments except control in semi-forcing culture was the highest between 15 and 30 cm soil depth at the 85 days after transplanting and between 0 and 15 cm soil depth after cucumber harvest. These results suggested that SCB 0.5N + Urea 0.5N can be used as an alternative N management for cucumber production in greenhouse if K accumulation is concerned.

Development of a Test Method for the Evaluation of DNA Damage in Mouse Spermatogonial Stem Cells

  • Jeon, Hye Lyun;Yi, Jung-Sun;Kim, Tae Sung;Oh, Youkyung;Lee, Hye Jeong;Lee, Minseong;Bang, Jin Seok;Ko, Kinarm;Ahn, Il Young;Ko, Kyungyuk;Kim, Joohwan;Park, Hye-Kyung;Lee, Jong Kwon;Sohn, Soo Jung
    • Toxicological Research
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    • v.33 no.2
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    • pp.107-118
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    • 2017
  • Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration ($IC_{50}$) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.

Construction of Retrovirus Vector System for the Regulation of Recombinant hTPO Gene Expression (재조합 hTPO 유전자의 발현 조절을 위한 Retrovirus Vector System의 구축)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Do-Hyang;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.161-167
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    • 2007
  • In this study, we constructed and tested retrovirus vectors designed to express the human thrombopoietin gene under the control of the tetracycline-inducible promoters. To increase the hTPO gene expression at him-on state, WPRE sequence was also introduced into retrovirus vector at downstream region of either the hTPO gene or the sequence encoding reverse tetracycline-controlled transactivator (rtTA). Primary culture cells (PFF, porcine fetal fibroblast; CEF, chicken embryonic fibroblast) infected with the recombinant retrovirus were cultured in the medium supplemented with or without doxycycline for 48hr, and induction efficiency was measured by comparing the hTPO gene expression level using RT-PCR, western blot and ELISA. Higher hPTO expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hTPO (in CEF) or rtTA(in PFF) gene. This resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

Direct Action of Relaxin on the Pig Mammary Glands to Promote Development of the Alveoli and Remodeling of the Extracellular Matrix (Relaxin Hormone의 직접젇인 작용에 의한 돼지 유선상피세포의 발달과 세포외간 기질의 분해개조 촉진)

  • Min, G.S.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.585-592
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    • 2004
  • Our previous studies demonstrated that relaxin in concert with estrogen promotes development of the mammary parenchyma during the last third of gestation in gilts, and the specific relaxin-binding sites were present in the mammary gland. This study was conducted to determine if relaxin-binding sites in the mammary gland were functional relaxin receptors. Three cycling cross-bred gilts were bilaterally ovariectomized on day 0 of the experiment. Beginning on day 15 and continuing through day 29 post-surgery, the gilt received an im. injection of estradiol benzoate at 12-hr intervals. Beginning on day 22 post-surgery, highly purified porcine relaxin was administered(lug/hr) into the left fourth mammary gland from the anterior end via miniature osmotic pump. Physiological saline was administered to the right fourth mammary gland. The gilt was sacrificed on day 29 post-surgery and histological characteristics of the mammary parenchyma were examined. The mammary glands treated locally with saline showed little, if any, lobulo-alveolar development, whereas the mammary glands treated with relaxin showed not only marked lobulo-alveolar development but also prominent secretions in the alveoli. The saline-treated glands were characterized by relatively dense and highly organized collagen fiber bundles. Whereas, in the relaxin-treated mammary glands, collagen fiber bundles were dispersed and loosely organized. In conclusion, relaxin-binding sites in the mammary gland are functional relaxin receptors and relaxin acts directly on the pig mammary gland to promote development of the alveoli and remodeling of the extracellular matrix.

Rainfall and Hydrological Comparative Analysis of Water Quality Variability in Euiam Reservoir, the North-Han River, Korea (북한강 의암호의 수질 변동성에 대한 강우·수문학적 비교분석)

  • Hwang, Soon-Jin;Sim, Yeon Bo;Choi, Bong-Geun;Kim, Keonhee;Park, Chaehong;Seo, Wanbum;Park, Myung-Hwan;Lee, Su-Woong;Shin, Jae-Ki
    • Korean Journal of Ecology and Environment
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    • v.50 no.1
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    • pp.29-45
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    • 2017
  • This study explored spatiotemporal variability of water quality in correspondence with hydro-meteorological factors in the four stations of Euiam Reservoir located in the upstream region of the North-Han River from May 2012 to December 2015. Seasonal effect was apparent in the variation of water temperature, DO, electric conductivity and TSS during the study period. Stratification in the water column was observed in the near dam site every year and vanished between August and October. Increase of nitrogen nutrients was observed when inflowing discharge was low, while phosphorus increase was distinct both during the early season with increase of inflowing discharge and the period of severe draught persistent. Duration persisting high concentration of Chl-a (>$25mg\;m^{-3}$: the eutrophic status criterion, OECD, 1982) was 1~2 months of the whole year in 2014~2015, while it was almost 4 months in 2013. Water quality of Euiam Reservoir appeared to be affected basically by geomorphology and source of pollutants, such as longitudinally linked instream islands and Aggregate Island, inflowing urban stream, and wastewater treatment plant discharge. While inflowing discharge from the dams upstream and outflow pattern causing water level change seem to largely govern the variability of water quality in this particular system. In the process of spatiotemporal water quality change, factors related to climate (e.g. flood, typhoon, abruptly high rainfall, scorching heat of summer), hydrology (amount of flow and water level) might be attributed to water pulse, dilution, backflow, uptake, and sedimentation. This study showed that change of water quality in Euiam Reservoir was very dynamic and suggested that its effect could be delivered to downstream (Cheongpyeong and Paldang Reservoirs) through year-round discharge for hydropower generation.

Effects of Compost Leachate on Growth and Yield of Leaf Lettuce in Hydroponic Culture (퇴비단 여과액비가 수경재배에서 상추의 생육과 수량에 미치는 영향)

  • Ryoo, Jong-Won
    • Journal of Animal Environmental Science
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    • v.15 no.1
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    • pp.51-58
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    • 2009
  • The compost leachate was dark-colored solution that leaches out of the bottom of the compost pile. The compost leachate was rich in nutrients and can potentially used in plant culture. In the organic production, commercial liquid fertilizer was used to insure the availability of nutrients during the formation of the yield. The cost of supplemental liquid fertilizer could be reduced by developing a fertilizer based on animal fertilizer. This experiment was conducted to investigate the effect of different combination of compost leachate and conventional inorganic solution in hydroponic culture for lettuce growth. Six different treatments were applied. The compost leachate(CL) and nutrient solution(NS) were mixed by six different mixing ratios of 0:100, 20:80, 40:60, 60:40%, 80:20 and 100%:0% based on nitrogen content. The chemical nutrient solution was the solution of National Horticulture Research Station for the growth of leaf lettuce. The concentration of nutrient solution was adjusted 1.5 mS/cm in EC. The compost leachate was low in phosphorus(P), calcium(Ca), magnesium(Mg), but rich in potassium(K). The plant height of lettuce treated with CL 20+NS 80% was similar with 100% NS of control plot. Plant height was highest in the plot of CL 20+NS 80%. The treatment of 100% compost leachate was lowest in the growth characteristics of leaf lettuce. Number of leaf was very low in 100% compost leachate compared with plot of chemical nutrient solution. In the beginning of growth stage, SPAD value was reduced in plot treated with CL100%, but CL20+NS80% plot was higher compared to 100% compost leachate. SPAD value of leaf lettuce leaves was decreased as the amount of CL was increased. The dry weight of lettuce were 107.4, 104.2g in plot of NS 100% and CL 20%+NS80%, respectively. The leaf number and plant weight were decreased at high application plots of compost leachate. The leaf lettuce showed lowest in the plot treated with 100% compost leachate, and the growth of lettuce severely decreased after application of 100% CL treatment. The results showed that compost leachate can be use as liquid fertilizer for the organic hydroponic production. The mixture solution of 20% of compost leachate and 80% of nutrient solution could be used as a nutrition solution in hydroponic culture of leaf lettuce. Our studies have shown that is possible to produce using compost leachate, although growth is slower than when using a conventional inorganic hydroponic solution.

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Effects of Organic Salts Fortified with Seaweed Components on Blood Pressure, Serum Minerals, and Hematochemicals in Spontaneously Hypertensive and Normotensive Rats (해조성분 강화염이 본태성 고혈압쥐와 정상혈압쥐의 혈압, 혈청 중 미네랄 함량 및 생화학적 특성에 미치는 영향)

  • Kim, Young-Myoung;Byun, Jee-Young;Han, Chan-Kyu;Sung, Ki-Seung;Namgung, Bae
    • Korean Journal of Food Science and Technology
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    • v.41 no.2
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    • pp.196-202
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    • 2009
  • The effects of the salts fortified with seaweed functional components on blood pressure, serum minerals, and hematochemicals in spontaneously hypertensive rats (SHR/NCrj) and normotensive rats (WKY/NCrj) were investigated. SHR and WKY rats were assigned to four groups, with 8 and 6 rats in each group: laver salt (A), fucoidan+laver high salt (B), fucoidan + laver low salt (C) and refined salt as a control (D). The final blood pressure (BP) of SHR and WKY species in contrast with reference BP were low in groups A and C as compared with control group. In terms of serum mineral content, $Na^+$ levels were similar in SHR and WKY, but $K^+$ levels were higher in the group B in SHR. Serum triglyceride levels were lower in groups A and C, but the levels of HDL- and LDL- cholesterol were significantly higher and lower in group A than those of controls, respectively (p<0.05). These results demonstrated that the salts fortified with laver might suppress blood pressure in rats, and also may improve mineral and lipid metabolism.

Determination of Carazolol and Azaperone in Livestock and Fishery Products Using Liquid Chromatography-tandem Mass Spectrometry (축수산물에서 LC-MS/MS를 이용한 카라졸롤 및 아자페론 분석)

  • Choi, Soo Yeon;Kang, Hui-Seung;Kim, Joohye;Cheon, So-Young;Jeong, Jiyoon;Cho, Byung-Hoon;Lee, Kang-Bong
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.176-184
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    • 2018
  • The aim of the present work was to develop simultaneous methods of quantification of carazolol, azaperone, and azaperol residues in livestock and fishery products using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Samples were extracted from beef, pork, chicken, egg, milk and shrimp using acetonitrile (ACN); while flat fish and eel were extracted using 80% ACN. For purification, ACN saturated n-hexane was used to remove fat composition. The standard calibration curves showed good linearity as correlation coefficients; $r^2$ was > 0.99. Average recoveries expressed were within the range of 67.9-105% for samples fortified at three different levels ($0.5{\times}MRL$, $1{\times}MRL$ and $2{\times}MRL$). The correlation coefficient expressed as precision was within the range of 0.55-7.93%. The limit of quantification (LOQ) was 0.0002-0.002 mg/kg. The proposed analytical method showed high accuracy and acceptable sensitivity based on Codex guideline requirements (CAC/GL71-2009). This method can be used to analyze the residue of carazolol, azaperone, and azaperol in livestock and fishery products.