Neural stem/precursor derived from pluripotent human embryonic stem cells (hESCs) has considerable therapeutic potential due to their ability to generate various neural cells which can be used in cell-replacement therapies for neurodegenerative diseases. However, production of neural cells from hESCs remains technically very difficult. Understanding neural-tube like rosette characteristic neural precursor cells from hESCs may provide useful information to increase the efficiency of hESC neural differentiation. Generally, neural rosettes were derived from differentiating hEBs in attached culture system, however this is time-consuming and complicated. Here, we examined if neural rosettes could be formed in suspension culture system by bypassing attachment requirement. First, we tested whether the size of hESC clumps affected the formation of human embryonic bodies (hEBs) and neural differentiation. We confirmed that hEBs derived from $500{\times}500\;{\mu}m$ square sized hESC clumps were effectively differentiated into neural lineage than those of the other sizes. To induce the rosette formation, regular size hEBs were derived by incubation of hESC clumps($500{\times}500\;{\mu}m$) in EB medium for 1 wk in a suspended condition on low attachment culture dish and further incubated for additional $1{\sim}2$ wks in neuroectodermal sphere(NES)-culture medium. We observed the neural tube-like rosette structure from hEBs after $7{\sim}10$ days of differentiation. Their identity as a neural precursor cells was assessed by measuring their expressions of neural precursor markers(Vimentin, Nestin, MSI1, MSI2, Prominin-1, Pax6, Sox1, N-cadherin, Otx2, and Tuj1) by RT-PCR and immunofluorescence staining. We also confirmed that neural rosettes could be terminally differentiated into mature neural cell types by additional incubation for $2{\sim}6$ wks with NES medium without growth factors. Neuronal(Tuj1, MAP2, GABA) and glial($S100{\beta}$ and GFAP) markers were highly expressed after $2{\sim}3$ and 4 wks of incubation, respectively. Expression of oligodendrocyte markers O1 and CNPase was significantly increased after $5{\sim}6$ wks of incubation. Our results demonstrate that rosette forming neural precursor cells could be successfully derived from suspension culture system and that will not only help us understand the neural differentiation process of hESCs but also simplify the derivation process of neural precursors from hESCs.
Journal of the Korean Society of Food Science and Nutrition
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v.38
no.11
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pp.1528-1534
/
2009
This study was designed to evaluate the anti-obesity and hypoglycemic effects of Gugija (Lycium chinense Mill) extracts in 3T3-L1 adipocytes. We investigated the $\alpha$-amylase and $\alpha$-glucosidase inhibitory activities of extracts from Gugija. Gugija was extracted by 70% EtOH and 80% MeOH and aqueous, respectively. A single oral dose of Gugija extract inhibited the increase of blood glucose levels significantly at 0, 30, 60, 90 and 120 min and decreased incremental response areas under the glycemic response curve. These results suggest that Gugija 70% EtOH extracts may delay carbohydrate digestion and reduce postprandial hyperglycemia. In addition, triglyceride content in 3T3-L1 adipocytes decreased at higher concentrations of Gugija 70% EtOH extract. Free fatty acid content in 3T3-L1 adipocytes was increased at higher concentrations of Gugija 70% EtOH extract. Also, glucose transporter 4 (GLUT4), the key insulin signaling pathway transcription factor, was remarkably increased by the Gugija 70% EtOH extract when compared to those of control cells in protein expression levels. Therefore, Gugija can be developed as an effective anti-obesity and hypoglycemic agent.
Park, Mirye;Kim, Z-Hun;Nam, Seung Won;Lee, Sang Deuk;Yun, Suk Min;Kwon, Dae Ryul;Lee, Chang Soo
Microbiology and Biotechnology Letters
/
v.48
no.2
/
pp.205-214
/
2020
Cyanobacteria are microorganisms which have important roles in the nitrogen cycle due to their ability to fix nitrogen in water and soil ecosystems. They also produce valuable materials that may be used in various industries. However, some species of cyanobacteria may limit the use of water resources by causing harmful algal blooms in water ecosystems. Many culture collection depositories provide cyanobacterial strains for research, but their systematic preservation is not well-developed in Korea. In this study, we developed a method for the cryopreservation of the cyanobacteria Trichormus variabilis (syn. Anabaena variabilis), using alginate microcapsules. Two approaches were used for the experiments and their outputs were compared. One of the methods involved the cryopreservation of cells using only a cryoprotectant and the other used the cryoprotectant within microcapsules. After cryopreservation for 35 days, cells preserved with both methods were successfully regenerated from the initial 1.0 × 105 cells/ml to a final concentration of 6.7 × 106 cells/ml and 1.1 × 107 cells/ml. Irregular T. variabilis shapes were found after 14 days of regeneration. T. variabilis internal structures were observed by transmission electron microscopy (TEM), revealing that lipid droplets were reduced after cryopreservation. The expression of the mreB gene, known to be related to cell morphology, was downregulated (54.7%) after cryopreservation. Cryopreservation using cryoprotectant alone or with microcapsules is expected to be applicable to other filamentous cyanobacteria in the future.
Ju-Hwi Park;Jongbeom Chae;Joon Ha Lee;Dongyup Hahn;Ju-Ock Nam
Journal of Applied Biological Chemistry
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v.66
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pp.447-454
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2023
In the environment in which humans live, there are various antigens that invade the human body and interfere with humans leading a healthy life, so the immune system recognizes the antigen then removes them through a complex mechanism. Macrophages are widely distributed immune cells involved in the innate immune system, and produce various immune modulators such as inducible nitric oxide synthase-induced nitric oxide, cyclooxygenase-2 induced prostaglandin E2 and proinflammatory cytokines such as tumor necrosis factor-alpha. On the other hand, Protaetia brevitarsis seulensis larvae are a type of edible insect that have emerged as an alternative to the future food supply problem. The immuno-modulatory effect through the activation of murine macrophage RAW264.7 cell via mitogen-activated protein kinases (MAPKs)/nuclear factor-kappa B (NF-κB) signaling pathways has been reported. Based on this report, in this study, we confirmed how the expression of immune modulators induced by Protaetia brevitarsis seulensis larvae extracts in RAW264.7 cells was changed by treatment with pharmacological inhibitors of toll-like receptor 4 (TLR4), MAPKs and NF-κB signaling pathways. As a result, reduction of immune modulators was confirmed in the c-Jun N-terminal kinase (JNK) inhibitor treatment group and NF-κB inhibitor treatment group among the Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell. Furthermore, in the TLR4 inhibitor-treated group, decreases in phosphorylation of JNK and NF-κB factors were confirmed in Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell, as well as decreases in immune modulators. This results suggest that Protaetia brevitarsis seulensis larvae activates RAW264.7 cells by the engagement of TLR4-JNK/NF-κB signaling pathway.
The mask dance drama Goseong Ogwangdae's first act, Mundung Bukchum. The story is about a man who was born to the upper class but contracted leprosy, an incurable disease, due to his ancestors' accumulated sin. Although he is suffering, he is able to triumph over the disease through sinmyeong (catharsis) that is expressed through the wordless act of Mundung Bukchum. This thesis focuses on Goseong Ogwangdae's Mundung Bukchum, particularly the meaning, costume, accompanying music, and process of pedagogical transmission. The chronological characteristics of Goseong Ogwangdae's Mundung Bukchum are as follows. First, as time goes on, the number of dance motions in Mundung Bukchum has increased and the motions used have become reified. Second, I address the small barrel drum used in Mundung Bukchum and check how, through the drum, the changed expression of Mundung is shown.In a 1965 video introduced Mundung is grasping the drum and stick and the dance appears to be made up only of humorous motions. Also in a 1969 video, "Mundung Gwangdae," from the start Mundung is grasping the drum and stick. In 1988 in a video we can see the same scene as today, with the drum and stick sitting in the center of the stage at the start of the dance. We can also confirm that the same fourteen dance motions used today are present. Third, we can also confirm the changes in Goseong Ogwangdae's signature motion, baegimsae as time goes on. Observing the video from 1965, baegimsae does not appear in Mundung Bukchum. In 2000 we can clearly see the baegimsae performed once to the left and to the right while Mundung is squatting. Comparing 1969 to 1988, there is no symmetry in the motions, baegimsae is not done to both left and right, but only in one direction. Watching the record from 2000, the baegimsae motion is performed just as it is now, with the body thrown forward with a sharp push off the ground. Fourth I confirmed how the music used to accompany Goseong Ogwangdae's Mundung Bukchum has changed over time. In 1965 according to records of Mundung Bukchum's appearance, the dance was accompanied by the taryeong rhythmic pattern played on the usual four percussion instruments (barrel drum, hourglass drum, large gong, small gong). In the 1969 records of Mundung Bukchum the accompanying music is gutgeori rhythmic pattern performed on the usual four percussion instruments-an obvious difference. In 1988 the music with Mundung Bukchum is gutgeori rhythmic pattern transitioning into jajinmori rhythmic pattern. In 2000 the music with Mundung Bukchum includes the percussion instruments as well as taepyeongso (double reed oboe) playing gutgeori and jajinmori rhythmic patterns.
Purpose: The interaction of the Fas: Fas ligand has been recognized to play an important role in radiation induced apoptosis. The purpose of this study was to investigate the role of Fas and Fas ligand mutations, in radiation-induced apoptosis in vivo. Materials and Methods: Mice with a mutation in the Fas ($C57BL/6J-Fas^{lpr}$) and its normal control (C57BL/6J) and the Fas ligand ($C3H/HeJ-Fas^{gld}$) and its normal control (C3H/HeJ), were used in this study. Eight-week old male mice were given whole body radiation. After irradiation, the mice were killed at various time intervals, and their spleens collected. Tissue sample was stained with hematoxylin-eosin, and the numbers of apoptotic cells scored. The regulating molecules of apoptosis including the p53, Bcl-2, Bax, $Bcl-X_L\;and\;Bcl-X_s$ genes were also analyzed by Western blotting. Results: With 2.5 Gy and 10 Gy of irradiation, the levels of apoptosis were lower in the $C57BL/6J-Fas^{lpr}\;and\;C3H/HeJ-Fas^{gld}$ mice than in the control mice (p<0.05). With the expression of apoptosis regulating molecules, the Bax was increased in both the C57BL/6J and C3H/HeJ mice in response to radiation; the peak levels of Bax in the C57BL6J and C3H/HeJ were 3 and 3.3-fold higher after 8hr, respectively. However the Bax was not increased in either the $C57BL/6J-Fas^{lpr}\;or\;C3H/HeJ-Fas^{gld}$mice. The p53, Bcl-X_L,\;Bcl-X_S$and Bcl-2 showed no significant changes in the $C57BL/6J-Fas^{lpr},\;C3H/HeJ-Fas^{gld}$, C57BL/6J and C3H/HeJ mice. Conclusion: The levels of radiation-induced apoptosis were lower in the lpr and gld, than the control mice, which seemed to be related to the level of Bax activation due to the radiation in the lpr and gld mice. This result suggests that Fas/Fas L plays an important role in radiation-induced apoptosis in vivo.
Journal of the Korean Society of Food Science and Nutrition
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v.32
no.8
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pp.1328-1336
/
2003
This study was done to investigate the effects of n-3, n-6 fatty acid and d-limonene on the hepatic membrane lipid composition, protein kinase C (PKC) and glutathione S-transferase (GST) activities in experimental rat hepatocarcinogenesis. Sprague-Dawley female rats were fed with two different types of dietary oil for 20 weeks. Corn oil (CO) and sardine oil (SO) were used at 15% by weight as a source of n-6 and n-3 fatty acid, respectively. One week after feeding, rats were intraperitoneally injected twice with a dose of diethylnitrosamine (DEN, 50 mg/kg body weight) and after 1 week 0.05% phenobarbital (PB) was provided with drinking water. Membrane fractional lipid composition showed that the content of cholesterol was higher in 50 group than CO group and also significantly decreased by d-limonene. The content of phospholipid was increased by carcinogen treatment but not affected by dietary oils or d-limonene. Membrane C/PL molar ratio was significantly decreased by d-limonene or carcinogen treatment in 50 groups but not in CO groups. Fatty acid composition was changed by dietary oils but not by carcinogen treatment or d-limonene. Cytosolic PKC activity was not significantly different by dietary oils, d-limonene or carcinogen treatment. However, membrane PKC activity was significantly increased by carcinogen treatment and decreased by d-limonene. Cytosolic GST activity was affected by d-limonene or carcinogen treatment in all dietary groups. These data indicate that dietary oils, d-limonene and carcinogen treatment can not change much membrane phospholipid composition. But membrane C/PL molar ratio was changed by carcinogen treatment and d -limonene although the effect was different between dietary oils. Therefore, it is suggested that different dietary oils and d-limonene can somewhat modulate the changes of membrane fluidity and activities of membrane bound enzymes like membrane associated PKC during carcinogenesis.
Journal of the Korean Institute of Intelligent Systems
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v.24
no.3
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pp.265-270
/
2014
Hearing-impaired people have difficulty in hearing, so it is also hard for them to learn letters that represent sound and text that conveys complex and abstract concepts. Therefore it has been natural choice for the hearing-impaired people to use sign language for communication, which employes facial expression, and hands and body motion. However, the major communication methods in daily life are text and speech, which are big obstacles for the hearing-impaired people to access information, to learn and make intellectual activities, and to get jobs. As delivering information via internet become common the hearing-impaired people are experiencing more difficulty in accessing information since internet represents information mostly in text forms. This intensifies unbalance of information accessibility. This paper reports web-based text-to-sign language translating system that helps web designer to use sign language in web page design. Since the system is web-based, if web designers are equipped with common computing environment for internet browsing, they can use the system. The web-based text-to-sign language system takes the format of bulletin board as user interface. When web designers write paragraphs and post them through the bulletin board to the translating server, the server translates the incoming text to sign language, animates with 3D avatar and records the animation in a MP4 file. The file addresses are fetched by the bulletin board and it enables web designers embed the translated sign language file into their web pages by using HTML5 or Javascript. Also we analyzed text used by web pages of public services, then figured out new words to the translating system, and added to improve translation. This addition is expected to encourage wide and easy acceptance of web pages for hearing-impaired people to public services.
Journal of the Korean Academy of Child and Adolescent Psychiatry
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v.11
no.1
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pp.110-123
/
2000
The short-term music therapy was performed for adolescents with conduct disorder admitted to Seoul National Mental Hospital for 3 months from Jun to September, 1998. This case study focused mainly on two female patients who participated regularly in the group music therapy. The music therapy process was divided into three phases;beginning, opening up, and closing. This music therapy session consisted of three parts;hello song as beginning, various musical activities, and sound & movement activity as closing. Free musical improvisation, song discussion, musical monodrama, and sound & movement were the mainly applied techniques. Free improvisation was used to enhance, motivate, identify and contain the adolescents' feelings and ideas. Song discussion was used to convey their thoughts and to support each other. Musical monodrama was used to make them have insights into interpersonal relationships. Sound & movement was used to enhance spontaneity. It made them explore their body and voice as an expressive medium. Throughout three months period of music therapy, patient A's communication skill, socialization, and behavior areas were assessed with improvement. She could use music as a symbolic form and was able to share her feelings about herself and her family. Patient B's self-expression and cognitive areas were assessed with improvement. She became more spontaneous and could verbalize her emotions during the group session. Music as a non-verbal and therefore often a non-threatening medium wherein so much can be expressed provided two female patients an atmosphere where a sense of trust may be regained.
Background : There have been many in vitro evidences that interleukin-4(IL-4) might be the most important cytokine inducing IgE synthesis from B-cells, and interferon-gamma(IFN-$\gamma$) might be a main cytokine antagonizing IL-4-mediated IgE synthesis. Recently some reports demonstrated that IFN-$\gamma$ might be used as a new therapeutic modality in some allergic diseases with high serum IgE level, such as atopic dermatitis or bronchial asthma. To evaluate the in vivo effect of IFN-$\gamma$ in bronchial asthma we tried a clinical study. Methods : Fifty bronchial asthmatics(serum IgE level over 200 IU/ml) who did not respond to inhaled or systemic corticosteroid treatment, and 17 healthy nonsmoking volunteers were included in this study. The CD 23 expressions of peripheral B-cells, the IL-4 activities of peripheral T-cells, the serum soluble CD23(sCD23) levels, and the superoxide anion(${O_2}^-$) generations by peripheral PMN were compared between bronchial asthmatics and normal subjects. The IL-4 activities of peripheral T-cells were analyzed by T-cell supernatant (T-sup)-induced CD23 expression from tonsil B-cells. In bronchial asthmatics the serum IgE levels and histamine $PC_{20}$ in addition to the above parameters were also compared before and after IFN-$\gamma$ treatment. IFN-$\gamma$ was administered subcutaneously with a weekly dose of 30,000 IU per kilogram of body weight for 4 weeks. Results : The ${O_2}^-$ generations by peripheral PMNs in bronchial asthmatics were higher than normal subjects($8.23{\pm}0.94$ vs $5.00{\pm}0.68\;nmol/1{\times}10^6$ cells, P<0.05), and significantly decreased after IFN-$\gamma$ treatment compared to initial values($3.69{\pm}0.88$ vs $8.61{\pm}1.53\;nmol/1{\times}10^6$ cells, P<0.05). CD23 expression of peripheral B-cells in bronchial asthmatics was higher than normal subjects($47.47{\pm}2.96%$, vs $31.62{\pm}1.92%$, P<0.05), but showed no significant change after IFN-$\gamma$ treatment. The serum sCD23 levels in bronchial asthmatics were slightly higher than normal subjects($191.04{\pm}23.3\;U/ml$ vs $162.85{\pm}4.85\;U/ml$), and 11(64.7%) of 17 patients showed a decreasing pattern in their serum sCD23 levels after IFN-$\gamma$ treatment. However the means of serum sCD23 levels were not different before and after IFN-$\gamma$ treatment. The IL-4 activities of peripheral T-cells in bronchial asthmatics were slightly higher than normal subjects($22.48{\pm}6.81%$ vs $18.90{\pm}2.43%$), and slightly increased after IFN-$\gamma$ treatment($27.90{\pm}2.56%$). Nine(60%) of 15 patients showed a decreasing pattern in their serum IgE levels after IFN-$\gamma$ treatment. And the levels of serum IgE were significantly decreased after IFN-$\gamma$ treatment compared to initial values ($658.67{\pm}120.84\;IU/ml$ vs $1394.32{\pm}314.42\;IU/ml$, P<0.05). Ten(83.3%) of 12 patients showed an improving pattern in bronchial hyperresponsiveness after IFN-$\gamma$ treatment, and the means of histamine $PC_{20}$ were significantly increased after IFN-$\gamma$ treatment compared to initial values ($1.22{\pm}0.29mg/ml$ vs $0.69{\pm}0.17mg/ml$, P<0.05). Conclusion : Our results suggest that IFN-$\gamma$ may be useful as well as safety in the treatment of bronchial asthmatics with high serum IgE level and that in vivo effects of IFN-$\gamma$ may be different from its in vitro effects on the regulations of IgE synthesis or the respiratory burst of PMN.
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