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Effects of BCG on the Absorptive Cells in the Appendix of the Mouse Implanted with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐의 막창자꼬리점막 흡수세포의 미세구조에 미치는 영향)

  • Lee, Woon-Woo;Park, Kyung-Ho;Kim, Myeong-Soo;Park, Dae-Kyoon;Ko, Jeong-Sik
    • Applied Microscopy
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    • v.37 no.3
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    • pp.157-166
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    • 2007
  • This experiment was performed to evaluate the ultrastructural responses of the absorptive cells in the appendix of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG (Bacillus Calmette-Guerin). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (experimental control group and BCG treated group). In the experimental groups, each mouse was inoculated with $1{\time}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.5mL of saline or BCG (0.5 mL/25gm B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8CFU$) were injected subcutaneously to the animals every other day. The day following the last injection, each mouse was sacrificed. Pieces of the tissue were taken from the appendix, prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. The ultrathin sections were stained with uranyl acetate and lead citrate. In the normal control, experimental control and BCG treated mice, general morphology of the absorptive cells of appendix were similar. But myelin figures and intramitochondrial dense granules were more frequently observed in the absorptive cells of BCG treated mice than normal control ones. Above results show that BCG did show slight ultrastructural alterations in the absorptive cell of the appendix. These results that BCG may slightly suppress function of the absorptive cells of the appendix.

Antioxidant and anti-inflammatory activities of extracts from Ledum palustre L. (백산차 추출물의 항산화 및 항염증 활성)

  • Kim, Se Gie
    • Food Science and Preservation
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    • v.24 no.7
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    • pp.1025-1033
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    • 2017
  • In this study, Ledum palustre L. was extracted by 4 different methods (LPW, hot water extraction; LPA, autoclave extraction; LPU, ultrasonification extraction; LPE, 70% ethanol extraction) and LPE was fractionated by using polarity difference of each solvent and used as 4 samples (LPE/H, the n-hexane layer; LPE/E, the EtOAc layer; LPE/B, the n-BuOH layer; LPE/W, the $H_2O$ layer). Antioxidant activities of Ledum palustre L. extracts were measured by DPPH and ABTS. As a result, the DPPH and ABTS radical scavenging showed high activities with LPE (82.3%, 99.8%) and LPE/E (91.8%, 99.6%) at the concentration of $1,000{\mu}g/mL$. The anti-inflammatory activities of LPE and LPE/E were measured by the inhibitory activity against NO, $PGE_2$, TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 production on LPS-stimulated Raw 264.7 macrophages. As a result of MTT assay, cell viabilities of LPE and LPE/E were more than 90% at $25{\mu}g/mL$. NO and $PGE_2$ productions were inhibited by LPE (NO: 50%, $PGE_2$: 70%) and LPE/E (NO: 57%, $PGE_2$: 73%) at the concentration of $25{\mu}g/mL$. The inhibition activities against TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production were 24%, 47% and 40% at the concentration of $25{\mu}g/mL$ of LPE. In particular, LPE/E showed 51%, 57% and 62% inhibition activities at the same concentration, respectively. From the above results, it can be concluded that $1,000{\mu}g/mL$ of LPE and LPE/E have the high antioxidant activities similar with Vitamin C, and $25{\mu}g/mL$, the low concetration of LPE and LPE/E have excellent anti-inflammatory activities. Therefore, if more research about anti-aging, whitening and antimicrobial activity of Ledum palustre L. extracts is carried out in the future, it will be possible to use them as effective materials for the prevention and treatment of inflammatory diseases and in the areas of functional foods and cosmetics.

Antioxidant Activities and Cell Viability against Cancer Cells of Adenophora remotiflora Leaves (모시잎의 항산화 효과 및 암세포주에 대한 세포 독성)

  • Kim, In-Sook;Park, Kwon-Sam;Yu, Hyeon-Hee;Shin, Mee-Kyung
    • Journal of the East Asian Society of Dietary Life
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    • v.19 no.3
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    • pp.384-394
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    • 2009
  • This study was performed to determine the antioxidative and anticancer effects of extracts from Adenophora remotiflora leaves. The antioxidative effects of the extracts were measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging activity and hemoglobin-induced linoleic acid oxidative inhibition assays. The results indicated that the extracts had stronger effects than the synthetic antioxidant BHT at the same concentration. The $SC_{50}$ values (50% radical scavenging effect on $1{\times}10^{-4}$ M DPPH) of the methanol fraction, water extract, and BHT were 47.5 ${\mu}g$/mL, 74.6 ${\mu}g$/mL and 102.2 ${\mu}g$/mL, respectively. In addition the $IC_{50}$ values (hemoglobin-induced linoleic acid oxidation inhibition) of the methanol fraction, water extract, and BHT were 120.8 ${\mu}g$/mL, 135.6 ${\mu}g$/mL, and 150.2 ${\mu}g$/mL, respectively. This research also assessed decreases in the survival of BNLcl2 cells (normal liver cells) by solvent fractions of the A. remotiflora leaf extracts at various concentrations (1, 5, 10, 25, 50, 100, 250, 500, 1,000, 2,000 ${\mu}g$/mL). The water extract did not decrease survival at any of the concentrations when compared to the control group. The hexane, ethyl acetate, and methanol fractions decreased survival as compared to the control group by inducing cell toxicity at a concentration of 1,000 ${\mu}g$/mL and above. Therefore, an anticancer activity experiment was conducted using concentrations below 500 ${\mu}g$/mL. At 500 ${\mu}g$/mL, the methanol fraction decreased A549 cell (human lung carcinoma cells) survival by 46% as compared to the control group, presenting the greatest effect against cell survival. All extracts showed greater anticancer activity in Hep G2 cells (human liver carcinoma cells) as compared to the A549 cells. For the Hep G2 cells, the methanol extract decreased survival by 28% as compared to the control group at the concentration of 500 ${\mu}g$/mL, thus restraining lung cancer cell growth.

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Studies on the Effects of Copper on the Lactate Dehydrogenase and Esterase Isozymes in Various Tissues of Carassius carassius (붕어(Carassius carassius)의 조직내 젖산수소이탈효소와 에스테라아제 아이소자임에 미치는 동의 영향에 관한 연구)

  • Lee, Choon-Koo;Choo, Il-Young
    • The Korean Journal of Zoology
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    • v.16 no.2
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    • pp.79-96
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    • 1973
  • In order to elucidate the effects of copper on Corassius carassius, the following were studied: 1) lactate dehydrogenase isozyme patterns by cellulose acetate electrophoresis, 2) LDH activity and copper effect on LDH enzyme system y spectrophotometry, 3) esterase isozyme patterns by agar thin layer electrophoresis, 4) hemoglobin patterns by starch gel electrophoresis, and 5) histological study. 1. There were two bands of LDH isozymes (LDH-3 and LDH-5) in the gill, three bands (LDH-2, LDH-4, and LDH-5) in the liver, and two bands (LDH-3 and LDH-4) in the muscle of the normal fish. The LDH-1 bond was not found in the above three tissues. When the fish were exposed to copper, LDH-3 appeared in the liver, LDH-5 in the muscle, but no new LDH band appeared in the gill. 2. The sepcific activities of the LDH were lowest in the gill and highest in the muscle of the normal fish, and they were gradually decreassed in the gill and highest in the muscle of the normal fish, and they were gradually decreased in the liver and mucle except in the gill from 1-day to 10-day exposure to copper. It indicates that LDH activities in the liver and muscle of the fish were inhibited by copper. 3. Through in vitro experiment, it is clear that the decrease of the LDH activities of the liver and muscle of the fish exposed to copper is mainly caused by the inhibition on the M-LDH in the fish. 4. The numbers of the esterase isozyme bands of the gill, liver, muscle, blood, brain, and kidney of the normal fish were 3, 6, 2, 2, 2, and 2 respectively, and these numbers were the same as those exposed to copper. The relative mobilities of the esterase bands in the gill, liver, blood, and kidney of the exposed group were different from those of the control. 5. There was one hemoglobin band on the anode in the normal fish. It seems that the nobility of hemoglobin band of the fish exposed to copper was slightly faster than that of the normal fish. 6. The normal gill lamellae of the fish consisted of centrally located pillar cells and a number of mucus cells. When the fish were exposed to copper, the epithelial layer was divorced first, disintegrated, and then destroyed completely. 7. The liver of the normal fish had prominent central veins, cords of hepatic cells, and sinusoids. When the fish were exposed to copper, numerous droplets of fat appeared in the cells around the central vein of the liver. It is assumed that the fatty droplets were accumulated by the lesion due to fatty metamorphosis of the liver caused by copper. 8. There was no histological difference between the muscle of the normal fish and that of the fish exposed to copper. 9. In the normal fish, the tubules of the kidney were surrounded by hemopoetic tissues. However, the kidney tissue of the fish exposed to copper received some damage on the proximal tubules. Since the tubule cells were reduced in height, the lumens of the tubules were enlarged. Consequently many proximal tubules exhibited some pink-stained granular casts and various stages of degeneration.

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Formation and Dissociation Kinetics of Zinc(II) Complexes of Tetraaza-Crown-Alkanoic Acids (Zinc(Ⅱ) Tetraaza-Crown-Allkanoic Acids 착물의 형성 및 해리 반응속도론)

  • Choi, Ki Young;Kim, Dong Won;Kim, Chang Suk;Park, Byung Bin;Choi, Suk Nam;Hong, Choon Pyo;Ryu, Hae Il
    • Journal of the Korean Chemical Society
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    • v.44 no.5
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    • pp.403-409
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    • 2000
  • The formation and dissociation rates of $Zn^{2+}$ Complexes with l,4,7,10-tetraaza-13,16-diox-acyclooctadecane-N,N',N",N'"-tetraacetic acid (1), 1,4,7,10-tetraaza-13,16- dioxacyclooctadecane-N,N',N",N'"-tetramethylacetic acid (2), and 1,4,7,10-tetraaza-13,16- dioxacyclooctadecane-N,N',N",N'"-tetrapropionic acid(3) have been measured by stopped-flow and conventional spectrophotometry. Observations were made at 25.0$\pm$0.1 $^{\circ}C$ and at an ionic strength of 0.10 M NaClO$_4$. The formation reactions of $Zn^{2+}$ ion with 1 and 2 took place by the rapid formation of an intermediate complex (ZnH$_3L^+$) in which the $Zn^{2+}$ ion is incompletely coor-dinated. This might then lead to be a final product in the rate-determining step.ln the pH range 4.76-5.76, the diprotonated (H2L2-) form is the kinetically active species despite of its low concentration. The stability con-stants (log$K_{(ZnH$_3$3$L^+$)}$) and specific water-assisted rate constants (koH) of intermediate complexes have been deter-mined from the kinetic data. The dissociation reactions of $Zn^{2+}$ complexes of 1,2, and 3 were investigated with $Cu^{2+}$ ions as a scavenger in acetate buffer. All complexes exhibit acid-independent and acid-catalyzed con-tributions. The effect of buffer and $Cu^{2+}$ concentration on the dissociation rate has also been investigated. The ligand effect on t dissociation rate of $Zn^{2+}$ complexes is discussed in terms of the side-pendant armsand the chelate ring sizes of the ligands.

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Spectrophotometric Determination of Aluminium Ion in Drinking Water by Flow Injection Analysis (흐름주입분석법에 의한 음용수 중 알루미늄 이온의 분광광도법 정량)

  • Choi, Yong-Wook;Jin, Jae-Young
    • Journal of the Korean Chemical Society
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    • v.44 no.5
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    • pp.422-428
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    • 2000
  • Optimum analytical conditions of the aluminium ion were established by flow injection analysis. Eriochrome Cyanine R(ECR) dye reacts with the aluminium ion at pH 6.0 to form a complex that exhibits maximum absorption at 535 nm. Reaction conditions including the mixing and the reaction coil length, the concentration and the pH of the buffer solutio, temperature, and injection loop volume were optimized to intro-duce this reaction into flow injection analysis. The results were as follows. A mixing coil length of 0.5 m and a reaction coil length of 4.0 m, the pH 6.0 and 1M of acetate buffer solution, the ECR concentration of 0.56 mM, the reaction temperature of 40$^{\circ}C$, the injection loop volume of 300${\mu}L$ were chosen as optimum conditions. Under these conditions the detection limit of the aluminiumion was less than 0.05 mg/L and the repeatability was better than 1%. A sampling frequency of 24 times for an hour was achieved. Interfering ions such as $F^-$, HP$O_4^{2-}$, $Fe^{2+}$, $Fe^{3+}$, $Mn^{2+}$, and other anions were tested, interference did not occur up to 1,000mg/L of ion concentration and up to 2,CO0mg/L of sulfate ion con-centration. This method was applied for the determination of aluminium ion in tap water and ground water of Jeonju and the Gochang area. The results showed that the aluminium residual in tap water of the Jeonju area was at a mean of 0.478mg/L and that in tap water of the Gochang area was at a mean of 0.278mg/L. Aluminium ion residual of the tap waters in the Jeonju area was higher level than that in the Gochang area. Aluminium residual in the ground water of the Jeonju area was 0.386 mg/L and was lower compared to 0.564 mg/L for the Gochang area.

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Antimutagenic and Cytotoxic Effects of Acer ginnala Max. Bark Extracts (신나무 껍질 추출물의 항돌연변이원성 및 세포독성 효과)

  • Oh Heung-Seok;Cui Cheng-Bi;Choi Hyung-Taek;Kim Soo-Hyun;Jeon Mi-Sun;Ham Seung-Shi
    • Food Science and Preservation
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    • v.11 no.4
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    • pp.550-556
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    • 2004
  • In the present study, we investigated the antimutagenic and cytotoxic effects of Acer ginnala Max. bark extract on S. typhimurium TA98, TA100 and cancer cell lines with Ames test and SRB assay, respectively. They were extracted with methanol and then fractionated using hexane, chloroform, ethyl acetate, butanol, and water to obtain the fractions. The inhibition rate of methanol ($200\;{\mu}g/plate$) of Acer ginnala Max. bark extract in the Salmonella typhimurium TA100 strain showed $83.3\%$ against the mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). In addition, the suppression of methanol extract with same concentration of in the Salmonella typhimurium TA98 and TA100 strains showed $80.3\%\;and\;92.7\%$ inhibition against 3-amino-1,4-dimethyl-5H-pyrido-(4,3-b)indol (Trp-P-1), respectively. The cytotoxicity effects of Acer ginnala Max. bark extract against the cell lines with human lung carcinoma (A549), human gastric carcinoma (AGS), human hepatocellular carcinoma (Hep3B) and human breast adenocarcinoma (MCF-7) were inhibited with the increase of the extract concentration. The treatment of 1.0 mg/mL Acer ginnala Max. bark methanol extract of methanol showed strong cytotoxicities of $77.3\%,\;90.4\%,\;88.9\%,\;and\;83.7\%$ against A549, AGS, Hep3B and MCF-7, respectively.

Fermentation and Quality Characteristics of Yakju with Addition of Chestnuts : Analysis of Raw Materials and Saccharification (쌀과 옥수수의 당화방법에 따른 밤 첨가 약주의 발효 및 품질 특성)

  • Huh, Chang-Ki;Seo, Jae-Sin;Kim, Yong-Doo
    • Food Science and Preservation
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    • v.15 no.4
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    • pp.512-517
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    • 2008
  • Fermentation characteristics of chestnut-added yakju prepared using various proportions of raw materials such as rice koji, rice flour, cornflour koji and cornflour were investigated. The pH of chestnut-added yakju prepared with cornflour koji and saccharified cornflour showed a higher value than that of chestnut-yakju prepared with rice koji and saccharified rice flour. The total acid content of chestnut-added yakju prepared with rice koji and saccharified rice flour was higher than that of chestnut-added yakju prepared using cornflour koji and sacharified cornflour. The reducing of sugar in chestnut-added yakju prepared with saccharified rice flour or saccharified cornflour was rapid at the first brewing stage, decreased dramatically after 2 days, and then decreased slowly after 5 days of fermentation. The value of L and a, the Hunter values, were high in chestnut-added yakju prepared with cornflour koji, and value b was high in chestnut-added yakju with rice koji. The content of iso-amyl alcohol was the highest of seven kinds of fusel oil found in chestnut-added yakju. Ethanol content increased to $17.6{\sim}18.2%$(v/v) after 8 days of fermentation. The content of lactic acid was the highest of all organic acids in the chestnut-added yakju. Sensory test results on chestnut-added yakju prepared with saccharifed corn flour showed that if rice flour is used as a sugar supplement for chestnut, the yakju prepared using koji had better flavor and taste. If cornflour was used in the preparation of chestnut-added yakju, the used of saccharified cornflour offered superior flavor and taste.

Comparison of the organic acids, fusel oil contents and antioxidant activities of Yakju with the additions of various rice cultivars (쌀 품종별 약주의 유기산, fusel oil 함량 및 항산화활성 비교)

  • Huh, Chang-Ki;Lee, Jung-Won;Kim, Yong-Doo
    • Food Science and Preservation
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    • v.20 no.3
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    • pp.365-371
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    • 2013
  • The organic acids, fuel oil contents and antioxidant activities of Yakju with the addition of rice varieties were investigated. The rice cultivars that were used in this study were Ilmi rice, Hopyung rice, Hopum rice, Hanarum rice, Anda rice and Indica rice. The study results are as follows: the pH values of Ilmi Yakju and Hanarum Yakju were 4.14 and 4.07, respectively, and those of the other Yakjus, 3.92~3.98. The total titratable acid content of the Yakju for which Indica rice was imported from Thailand was highest among the samples. The reducing sugars content of the Yakju for which Indica rice was used was higher than that of the other samples. The ethanol content of the Hanarum Yakju was higher than that of the other Yakjus, and the lowest ethanol content was found for the Indica Yakju. The major organic acid component of the Yakju was lactic acid. The total organic acid content was highest in the Indica rice Yakju. The amounts of the following main fusel oils were highest, in the following order: 2-butyl alcohol, 2-pentanol, N-propyl alcohol, acetaldehyde, ethyl acetate, methyl alcohol and 2-methyl-1-propyl alcohol. The DPPH free radical scavenging activity of the Indica rice Yakju was much higher than that of the Ilmi rice Yakju and the Hanarum rice Yakju. Also, the SOD activity of the Indica rice Yakju was strongest among all the samples. The total polyphenol content of the Indica rice Yakju was the highest among all the samples.

Development and Validation of the Determination of Sorafenib in Human Plasma using Tandem Mass Spectrometry Coupled with Liquid Chromatography (고속액체크로마토그래피 텐덤질량분석기법을 이용한 사람 혈장 내 소라페닙 농도분석법의 개발 및 검정)

  • Park, Daejin;Lee, Sunggon;Kim, Woomi
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1456-1462
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    • 2012
  • Sorafenib is a multikinase inhibitor and an oral anticancer drug approved for the treatment of patients with advanced renal cell carcinoma and those with unresectable hepatocellular carcinoma. The purpose of this study was to develop an efficient method of the determination of sorafenib in human plasma using tandem mass spectrometry coupled with liquid chromatography (LC/MS/MS) and validate the method by the guidelines of the Korean Food and Drug Administration (KFDA). Plasma samples ($100{\mu}l$) were added with chlorantraniliprole as an internal standard and then mixed with the 0.1% formic acid-containing extraction solution composed of isopropyl alcohol and ethyl acetate (1:4, v/v). After centrifugation, the supernatant was concentrated at $45^{\circ}C$ under negative pressure and centrifugal force. The residue was reconstituted with a mobile phase and injected into the HPLC instrument using a reverse phase Waters XTerra$^{TM}$ C18 column (particle size $3.5{\mu}m$). Liquid chromatography was carried out within the run time of 5 min using a mobile phase composed of buffer (0.1% formic acid and 10 mM ammonium formate), methanol, and acetonitrile (1:6:3, v/v/v). The analytes were monitored by tandem mass spectrometry in the multiple reaction monitoring method programmed to detect sorafenib at 'm/z 465.2 ${\rightarrow}$ 252.5' and chlorantraniliprole at 'm/z 484.4 ${\rightarrow}$ 286.2' with positive electrospray ionization mode ($ES^+$). The result showed the proper linearity ($r^2$ > 0.99) over the range of 2,000-5,000 ng/ml with good accuracy (90.7-103.9%) and precision (less than 10%). The newly developed method using LC/MS/MS was validated by the guideline of KFDA and identified as more sensitive compared to the previous methods.