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Differentiation and Apoptosis of the Mammalian Embryo and Embryonic Stem Cells(ESC): I. Establishment of Mouse ESC and Induction of Differentiation by Reproductive Hormones (포유동물의 배아 및 기간세포의 분화와 세포사멸 기작: I. 생쥐 배아줄기세포의 확립과 분화유도에 미치는 생식호르몬의 영향)

  • 성지혜;윤현수;이종수;김철근;김문규;윤용달
    • Development and Reproduction
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    • v.6 no.1
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    • pp.55-66
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    • 2002
  • Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

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Geochemical Exploration Technics in the Pungchon Limestone Area (풍촌 석회암지대 탐사에 적용될 새 지화학탐사법 연구)

  • Moon, Kun Joo
    • Economic and Environmental Geology
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    • v.23 no.4
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    • pp.369-381
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    • 1990
  • Most of significant ore deposits in South Korea such as the Sangdong W - Mo, the Yeonhwa Pb-Zn and the Geodo Cu-Fe skarn ore deposits occur at the southern limb of the Hambaeg syncline in the Taebaeg Basin. The mineralization took place in the interbedded limestone of the Myobong Formation and the Pungchon limestone of the Great Limestone Group of the Cambrian age, generally striking E-W and dipping 25-30 degrees north. There are no outcrops of the skarn-type orebody at the northern limb of the syncline. In order to find a clue of a possible hidden orebody localized at the limestones in the northern limb, a lithogeochemical exploration by using carbon isotope and some elements such as Si, Ca, Fe and Al at the Sangdong Mine area has been attempted as for a modelling study. For this study, 45 samples from the Pungchon limestone which do not show any megascopic indication of mineralization have been taken in both the mineralized zone and the unminerallized zone at the Sangdong Mine area. Analytical data show that there are big differences in the contents of CaO and $Al_2O_3$ between the Pungchon limestone of the mineralized zone and that of the unmineralized zone. Carbon isotope data exhibit that ${\delta}^{13}C$ values of the Pungchon limestone in the mineralized zone are highter than those in the unmineralized zone. The difference in the analytical values of CaO, $Al_2O_3$ and the carbon isotope between the mineralized and the unmineralized zones is as follows ; Unminerallized zone Mineralized zone CaO 51.3% 43.5% $Al_2O_3$ 0.6% 2.4% ${\delta}^{13}C$ -0.39 permil -0.56 permil $Fe_2O_3$ 0.9% 1.4% $SiO_2$ 3.0% 2.4% The decrease in the Si content of the Pungchon limestone in the mineralized zone is contrary to the result of the previous study (Moon, 1987). On the basis of identification of the increase in the Al content of the limestone in the mineralized zone, it could be deduced that the decrease in the Si content of the Pungchon limestone might be due to the result of increase in the alteration products mainly occurred along fracture-system such as joint cracks or minor faults and that the phenomena shown by the Si and Al content in the mineralized zone might be derived from the thermal effect of granite extended mineralizing activity to the overlied limestone on the surface. Higher mean values of Fe and Al as well as lower mean values of carbon content and the ${\delta}^{13}C$ than mean values of those in the Pungchon limestone at the northern limb of the Hambaeg Syncline may be applicable in exploration for blind orebodies.

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The Origin and Evolution of the Mesozoic Ore-forming Fluids in South Korea: Their Genetic Implications (남한의 중생대 광화유체의 기원과 진화특성: 광상 성인과의 관계)

  • Choi, Seon-Gyu;Pak, Sang-Joon
    • Economic and Environmental Geology
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    • v.40 no.5
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    • pp.517-535
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    • 2007
  • Two distinctive Mesozoic hydrothermal systems occurred in South Korea: the Jurassic/Early Cretaceous(ca. $200{\sim}130$ Ma) deep-level ones during the Daebo orogeny and the Late Cretaceous/Tertiary(ca. $110{\sim}45$ Ma) shallow hydrothermal ones during the Bulgugsa event. The Mesozoic hydrothermal system and the metallic mineralization in the Korean Peninsula document a close spatial and temporal relationship with syn- to post-tectonic magmatism. The calculated ${\delta}^{18}O_{H2O}$ values of the ore-forming fluids from the Mesozoic metallic mineral deposits show limited range for the Jurassic ones but variable range for the Late Cretaceous ones. The orogenic mineral deposits were formed at relatively high temperatures and deep-crustal levels. The mineralizing fluids that were responsible for the formation of theses deposits are characterized by the reasonably homogeneous and similar ranges of ${\delta}^{18}O_{H2O}$ values. This implies that the ore-forming fluids were principally derived from spatially associated Jurassic granitoids and related pegmatite. On the contrary, the Late Cretaceous ferroalloy, base-metal and precious-metal deposits in the Taebaeksan, Okcheon and Gyeongsang basins occurred as vein, replacement, breccia-pipe, porphyry-style and skarn deposits. Diverse mineralization styles represent a spatial and temporal distinction between the proximal environment of subvolcanic activity and the distal to transitional condition derived from volcanic environments. The Cu(-Au) or Fe-Mo-W deposits are proximal to a magmatic source, whereas the polymetallic or the precious-metal deposits are more distal to transitional. On the basis of the overall ${\delta}^{18}O_{H2O}$ values of various ore deposits in these areas, it can be briefed that the ore fluids show very extensive oxygen isotope exchange with country rocks, though the ${\delta}D_{H2O}$ values are relatively homogeneous and similarly restricted.

Autoradiographic Studies on the Inhibitory Effect of Dibutyryl Cyclic AMP on Mouse Oocyte Maturation in Vitro (Dibutyryl Cyclic AMP가 생쥐여포난자의 성숙에 미치는 억제효과에 관한 자기방사법적 연구)

  • Choi, Choon-Keun
    • Applied Microscopy
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    • v.7 no.1
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    • pp.21-43
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    • 1977
  • This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

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HACCP Application of Instant Ablactation Baby Food Processing (즉석 이유식 제조시 HACCP의 적용)

  • 이정규;노완섭
    • The Korean Journal of Food And Nutrition
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    • v.14 no.5
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    • pp.457-466
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    • 2001
  • In this study. HACCP system was applied to improve the quality of instant ablactation baby food with rising sanitary problem among SunSig(powder of roasted grains and vegetables) . It took about 9min 30 second to produce and the temperature status of raw materials were 30.9∼31.8$\^{C}$. Raw material and production phase goods had average 6.2 in pH and 0.23∼0.63 in water activity. The average number of total aerobic bacteria(TAB) and coliform were 1.4$\times$10$^4$ CFU/g, 5.5$\times$10 CFU/g in raw materials. And the number became 8.7$\times$10$^2$ CFU/g, 6.9$\times$10 CFU/g after 1st grinding step. The distributions of TAB and coliform in equipments were 3.8$\times$10$^4$ CFU/㎠, 8.0$\times$10 CFU/㎠ on the average. According to storage temperature, variations of number of TAB coliform were slight at 4$\^{C}$ but increased at 20$\^{C}$ and 30$\^{C}$ . Therefore following subjects can be issued on the basis of results. Minimize the storage time and keep at cool temperature. 1st and 2nd grinders should be managed hygienically. Establish the heating step during the production of instant action baby food. Instant ablactation baby food produced should be stored at cool and freeze condition.

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Effects of Isoflavone Supplementation on Lipid Profiles and Antioxidant Systems in Rats Fed with Cholesterol Diet (고콜레스테롤 식이를 섭취한 흰쥐에서 이소플라본 보충이 혈청 지질패턴 및 항산화체계에 미치는 영향)

  • Kim, Soo-Yeon;Kim, Soon-Young;Chung, Chung-Eun;Yoon, Sun;Park, Jung-Hwa
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1683-1690
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    • 2010
  • There is an increasing interest in the potential of isoflavone in reducing the risk of cardiovascular disease and cancer, however, although several effects of isoflavone as a component of soy protein are well established, the hypolipidemic and antioxidant effects of purified isoflavone are still controversial. This study was to investigate the effects of isoflavone on serum lipid profiles and antioxidant status in rats. 7-week old male Sprague Dawley rats were fed one of the following diets for 8 weeks: basal diet (B), basal+0.3% isoflavone (BI), basal+0.5% cholesterol (BC), or basal+0.3% isoflavone +0.5% cholesterol (BIC). Two-way ANOVA was used to test the effects of dietary isoflavone and cholesterol supplementation and their interaction on variables. Serum lipid profiles and total antioxidant status (TAS) were examined spectrophotometrically. Degree of serum lipid peroxidation was measured by malondialdehyde (MDA) assay. The activities of serum antioxidant enzymes (GSH-Px, total-SOD) was determined. Levels of serum total cholesterol, VLDL+LDL-cholesterol and Atherogenic index were significantly lower in BI than those levels in group B (p=0.0002, p<0.0001, and p=0.0042, respectively). Serum total antioxidant status (TAS) levels were significantly higher, in both isoflavone supplemented groups (BI, BIC) compared to those levels in each control group (B, BC) (p<0.0001). Activity of total-SOD was significantly higher in BI compared to the activities in group B (p=0.0317). There was no interaction between isoflavone and cholesterol supplementation. In conclusion, isoflavone supplementation showed positive effects on the serum lipid profiles and total antioxidant activities in both conditions, either when fed a diet with or without cholesterol. These effects of isoflavone were independent of cholesterol supplementation.

Protective Effect of Isoflavone, Genistein from Soybean on Singlet Oxygen Induced Photohemolysis of Human Erythrocytes ($^1O_2$으로 유도된 사람 적혈구의 광용혈에 있어서 대두의 아이소플라본인 제니스테인의 보호작용)

  • Park, Soo-Nam
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.510-518
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    • 2003
  • Protective effects of natural components including genistein (4',5,7-trihydroxyisoflavone) from Glycine max MERRILL on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. Genistein $(10{\sim}100\;{\mu}m)$ suppressed photohemolysis in a concentration-dependent manner, and was more effective than the lipid peroxidation chain blocker, ${\alpha}$-tocopherol (Vit. E). Glycoside of genistein, genistin, the water-soluble antioxidant, L-ascorbate, and the iron chelator, myo-inositol hexaphosphoric acid dodecasodium salt (sodium phytate) did not exhibit protective effect against photohemolysis. L-Ascorbate and sodium phytate stimulated photohemolysis at high concentration $(500\;{\mu}m)$. ${\alpha}$-Carotene 3,3'-diol (lutein), a singlet oxygen $(^1O_2)$ quencher, exhibited pronounced protective effect, an indication that $^1O_2$ is important in photohemolysis sensitized by rose-bengal. Reactive oxygen scavenging activities $(OSC_{50})$ of natural antioxidants including genistein on reactive oxygen species (ROS) generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay were in the order of sodium phytate > L-ascorbate > ${\alpha}$-tocopherol > genistein > genistin. $OSC_{50}$ value of genistein, genistin, ${\alpha}$-tocopherol, L-ascorbate, and sodium phytate were 41.0, 109.0, 9.0, 5.2, and $0.56{\mu}m$ respectively. The order of free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity $(FSC_{50})$ was L-ascorbate > ${\alpha}$-tocopherol > genistein > genistin. These results indicate that genistein can function as an antioxidant in biological systems, particularly skin exposed to solar UV radiation by scavenging $^1O_2$ and other ROS, and to protect cellular membranes against ROS.

The Change of Cortical Activity Induced by Visual Disgust Stimulus (시각혐오자극으로 유발된 대뇌 피질 활성도 변화)

  • Jung, Wook;Park, Doo-Heum;Yu, Jae-Hak;Ryu, Seung-Ho;Ha, Ji-Hyeon;Shin, Byoung-Hak
    • Sleep Medicine and Psychophysiology
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    • v.20 no.2
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    • pp.75-81
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    • 2013
  • Objectives: There are a lot of studies that analyze the interaction between the emotion of disgust and the functional brain images using fMRI and PET. But studies using sLORETA (standardized low resolution brain electromagnetic tomography) almost do not exist. The aim of this research is to explore the relationship of the emotion of disgust and the cortical activation using sLORETA analysis. Methods: Forty five healthy young adults ($27.1{\pm}2.6$ years) participated in the study. While they were watching 4 neutral images and 4 disgusting images associated with mutilation selected from the international affective picture system (IAPS), participants' EEGs were taken for 30 seconds per one picture. Through these obtained EEG data, sLORETA analysis was performed to compare EEGs associated with neutral and negative images. Results: During looking for visual disgusting stimulus, all participants reported unpleasantness, arousal and stress. In sLORETA analysis, the decrease of current density in theta wave was shown at left frontal superior gyrus (BA10) and middle gyrus (BA10, 11). This voxel cluster consists of a total of 11 voxels and the threshold of t value indicating statistically significant decreases in the current density (p<0.05) was -1.984. There were no differences between male and female in the degree of being disgusted by the stimuli. Conclusion: This finding may suggest that the activation of dorsolateral prefrontal cortex might be associated with regulating disgust emotion.

Antioxidant and Cellular Protective Activities of Ecklonia cava Extracts against Reactive Oxyegen Species (감태(Ecklonia cava) 추출물의 항산화 및 세포보호 활성)

  • Yoo, Cha Young;Kim, Si Yun;Park, Jung Won;Sung, Soo An;Kim, Da Ae;Park, Jee Hyun;Xuan, Song Hua;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.3
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    • pp.287-294
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    • 2015
  • In this study, we investigated the antioxidative effects of brown seaweed Ecklonia cava extract and its subfractions. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction of E. cava. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of ethyl acetate fraction ($FSC_{50}=6.98{\mu}g/mL$) and aglycone fraction ($7.03{\mu}g/mL$) are similar to that of (+)-${\alpha}$-tocopherol ($8.98{\mu}g/mL$) which is a reference control. Reactive oxygen species (ROS) scavenging activity (total antioxidant capacity, $OSC_{50}$) of the aglycone fraction ($OSC_{50}=14.48{\mu}g/mL$) on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay was the strongest among all extract and fractions. However, all samples showed lower antioxidant activities than that of L-ascorbic acid ($6.88{\mu}g/mL$) known as a powerful antioxidant. The protective effect of 50% ethanol extract on the $^1O_2$-induced cellular damage of human erythrocytes was dependent on the concentration from 5 to $50{\mu}g/mL$. Both ethyl acetate fraction and aglycone fraction showed strong cellular protective activities at $10{\mu}g/mL$, where the cellular protective effects (${\tau}_{50}$) of each fraction were recorded 442.0 min and 539.9 min, respectively. Three kinds of extract/fractions of E. cava showed much greater cellular protective activities at $10{\mu}g/mL$ than that of liposoluble antioxidant (+)-${\alpha}$-tocopherol (40.6 min) which is a reference control. These results suggest E. cava extracts and its fractions can be applied as an antioxidant ingredient in a field of cosmetics.

Effects of 20(S)-Protopanaxadiol and 20(S)-Protopanaxatriol on the Inflammatory Mediators Release from the Activated Mast Cells (20(S)-Protopanaxadiol 및 20(S)-Protopanaxatriol이 활성화된 비만세포로부터의 염증 매개체 유리에 미치는 영향)

  • Ro, Jai-Youl;Han, Yong-Nam;Choi, Kwang-Tae;Lee, Chang-Ho
    • Journal of Ginseng Research
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    • v.33 no.4
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    • pp.316-323
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    • 2009
  • Ginseng saponins have various pharmacological effects on the immune system. 20(S)-protopanaxadiol (PPD) and 20(S)-protopanaxatriol (PPT) are the species of ginseng saponin metabolites that are formed by human intestinal bacteria and detected in circulation. The effects of PPD and PPT on the inflammatory mediator release from the activated mast cells were tested. Histamine release was evaluated in activated guinea pig lung mast cells, and the secretion of interleukin-4 (IL-4), interleukin-8 (IL-8), and the tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) was assessed in an HMC-1 cell after treating it with ginseng saponin metabolites. The results are as follows. PPT, at its maximum concentration of $100\;{\mu}M$, completely abolished the secretion of IL-4 from the PMA-stimulated HMC-1 cell. It also inhibited IL-8 secretion from the same cells by about 40-50% of the PMA-treated DMSO control. PPD, at its maximum concentration of $100\;{\mu}M$, showed a tendency to induce histamine release from the guinea pig lung mast cells. It inhibited the secretion of IL-4 (by 89% of the PMA-treated DMSO control) in the PMA-stimulated HMC-1 cell, but did have a significant effect on the IL-8 release from the same cell. Both PPD and PPT showed no effects, however, on the release of TNF-${\alpha}$ from the PMA-stimulated HMC-1 cell. These results suggest that PPD and PPT are from the ginseng metabolites that are responsible for the immunomodulating activity of ginseng extracts when they are taken orally.