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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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A Study of the Representation in the Elementary Mathematical Problem-Solving Process (초등 수학 문제해결 과정에 사용되는 표현 방법에 대한 연구)

  • Kim, Yu-Jung;Paik, Seok-Yoon
    • Journal of Elementary Mathematics Education in Korea
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    • v.9 no.2
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    • pp.85-110
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    • 2005
  • The purpose of this study is to examine the characteristics of visual representation used in problem solving process and examine the representation types the students used to successfully solve the problem and focus on systematizing the visual representation method using the condition students suggest in the problems. To achieve the goal of this study, following questions have been raised. (1) what characteristic does the representation the elementary school students used in the process of solving a math problem possess? (2) what types of representation did students use in order to successfully solve elementary math problem? 240 4th graders attending J Elementary School located in Seoul participated in this study. Qualitative methodology was used for data analysis, and the analysis suggested representation method the students use in problem solving process and then suggested the representation that can successfully solve five different problems. The results of the study as follow. First, the students are not familiar with representing with various methods in the problem solving process. Students tend to solve the problem using equations rather than drawing a diagram when they can not find a word that gives a hint to draw a diagram. The method students used to restate the problem was mostly rewriting the problem, and they could not utilize a table that is essential in solving the problem. Thus, various errors were found. Students did not simplify the complicated problem to find the pattern to solve the problem. Second, the image and strategy created as the problem was read and the affected greatly in solving the problem. The first image created as the problem was read made students to draw different diagram and make them choose different strategies. The study showed the importance of first image by most of the students who do not pass the trial and error step and use the strategy they chose first. Third, the students who successfully solved the problems do not solely depend on the equation but put them in the form which information are decoded. They do not write difficult equation that they can not solve, but put them into a simplified equation that know to solve the problem. On fraction problems, they draw a diagram to solve the problem without calculation, Fourth, the students who. successfully solved the problem drew clear diagram that can be understood with intuition. By representing visually, unnecessary information were omitted and used simple image were drawn using symbol or lines, and to clarify the relationship between the information, numeric explanation was added. In addition, they restricted use of complicated motion line and dividing line, proper noun in the word problems were not changed into abbreviation or symbols to clearly restate the problem. Adding additional information was useful source in solving the problem.

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The Increase of Apoptotic Neutrophils and Phagocytic Macrophage by Germanium in Acute Lung Injury Induced by Lipopolysaccharide (LPS에 의한 급성 폐손상에서 게르마늄에 의한 호중구 세포사와 큰포식세포의 포식능 증가)

  • Lee, Yoon-Jeong;Cho, Hyun-Gug;Jeune, Kyung-Hee
    • Applied Microscopy
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    • v.38 no.4
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    • pp.293-306
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    • 2008
  • The essential factor of acute respiratory distress syndrome (ARDS), an acute lung injury accompanied commonly by sepsis syndrome is accumulation of neutrophils in lung tissue. The study attempted to confirm whether a lung injury would be decreased with the anti-inflammatory effect of germanium by the treated germanium prior to the development of ARDS and whether nitric oxide influence in suppressing a lung injury. Test groups were divided in the following structure for experiment; CON that has been administered with sodium chloride to airway, LPS administered with endotoxin for 5 hours in the same amount and 5 hours of endotoxin administered Ge+LPS following 1 hours of pre-treated germanium. The result of a test using experimental animals, infilteration of neutrophils (p<0.001) in bronchoalveolar lavage fluid (BALF) was significantly decreased, the structure of lung tissue was preserved relatively well, and much neutrophils with distinct positive were observed on tunel staining which showed increase of apoptotic neutrophils in the pre-treated germanium group compare to the endotoxin administrated group. In observation of ultrastructural changes of cell in BALF, phagocytic alveolar macrophage was increased in alveolar space, the nucleus of most engulfed neutrophils were condensed, and some apoptosis neutrophils appears to be DNA fragmentation and effacement of cellular organelles were found in intercellular matrix in the pre-treated germanium group. However, the nitric oxide showed increase in all the groups excluding CON, and the nitric oxide effect such as degranulation diminishing of mast cells and apoptosis increase of neutrophils in the pre-treated group only. The situation appears that there was change in internal environment of the experimental animal by the pre-treated germanium before the nitric oxide is produced and the anti-inflammatory effect activated the pre-processed germanium by nitric oxide which activated following the change. Therefore, the nitric oxide created from macrophage in accordance with the pre-treated germanium appears to influence in alleviating a lung injury. Accordingly, acute lung injury is alleviated by the anti-inflammatory effect of germanium such as inhibition of neutrophils migration, induction of neutrophil apoptosis and increase of phagocytic function of phagocyte, and the nitric oxide produced from activated macrophage by germanium would influence in suppressing a lung injury.

Survey on Quality of Hospice.Palliative Care Programs in Korea (한국 호스피스.완화의료기관 실태 조사)

  • Yun, Young-Ho;Choi, Eun-Sook;Lee, In-Jeong;Rhee, Young-Sun;Lee, Jung-Suk;You, Chang-Hoon;Kim, Hyun-Sook;Paek, Yu-Jin
    • Journal of Hospice and Palliative Care
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    • v.5 no.1
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    • pp.31-42
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    • 2002
  • Purpose : The purpose of this study was to evaluate the present status of hospice palliative care programs in Korea as a basic database for standardization of hospice palliative care. Method : The data was collected from July to October, 2001. The instrument used for this study was the questionnaires which was consisted of the general characteristics of organization, recipient of service, manpower, contents of service, financial conditions and facilities. Sixty-four hospice palliative care programs answered the questionnaires, confirmed by telephone. Results : They were 40 hospital-based hospice palliative care programs and 24 nonmedical hospice palliative care programs. 11 Hospital-based hospice palliative programs have isolated unit or hospital affiliated free standing hospice. 6 Non-hospital hospice palliative programs have a free standing hospice. Major subjects of hospice palliative program were terminal cancer patients but patients with non-terminal illness were also included. Only 24 of 64 hospice palliative programs had all of the essential professionals : physicians, nurses, social workers, and clergies. Home hospice palliative care programs have a referral system in hospital based (89.7%) and nonmedical programs (73.7%). 24hr hospice are were provided in 26 hospital-based (65.0%) and 9nonmedical programs (37.5%). There were rooms for family in half of hospital-based programs. 73.9% of hospice palliative care programs have financial problems. 62.0% of Hospice palliative care programs need financial support from government. Conclusion : 64 Hospice palliative care programs provided hospice palliative services but had many problems in manpower, quality of care and facility. For improving the quality of terminal patients' life and promoting the cost effectiveness of health care resources, it is necessary to consider the standardization and institutionalization of hospice palliative care.

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Effects of Aromatherapy on Depression, Anxiety and the Autonomic Nervous System in Breast Cancer Patients Undergoing Adjuvant Radiotherapy (방사선 요법을 받는 유방암 환자에게 적용한 아로마 요법이 우울, 불안 및 자율신경계 반응에 미치는 효과)

  • Yun, Sun-Hee;Cha, Jung-Hee;Yoo, Yang-Sook;Kim, Yeong-In;Chung, Su-Mi;Jeong, Hea-Lim
    • Journal of Hospice and Palliative Care
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    • v.15 no.2
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    • pp.68-76
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    • 2012
  • Purpose: The study was designed to verify effects of aromatherapy on depression, anxiety and the autonomic nervous system in breast cancer patients who are undergoing adjuvant radiotherapy. Methods: Data were collected from November 2006 through March 2007 at the C university hospital in Seoul. The study included 33 patients and they were assigned to three groups. Group I had aroma oil inhalation for 2 minutes per time, three times a day for six weeks whereas Group II and a control group inhaled aroma oil for 2 minutes per time, once a day for six weeks. For Groups I and II, a mixture of lemon, lavender, rosewood and rose essential oils were used while control group inhaled tea tree oil. Results: Depression was significantly decreased in patients in the experimental groups only. In Groups I and II patients, anxiety level was lower than that in control patients. Patients in Groups I and II also showed stronger physical resistance to stress than control group patients. Conclusion: Aromatherapy should be considered as a method that can significantly decrease depression in breast cancer patients who are undergoing adjuvant radiotherapy.

Analysis of Volatile Flavor Compounds from the Leaves of Eucommia ulmoides (두충잎의 휘발성 향기성분 분석)

  • 이미순;정미숙
    • Korean journal of food and cookery science
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    • v.17 no.4
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    • pp.359-366
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    • 2001
  • This study was performed to analyze the volatile flavor compounds of Eucommia ulmoides leaves as influenced by harvesting time and drying method. Essential oils of fresh, air-dried and freeze-dried leaves of Eucommia ulmoides were extracted by SDE(simultaneous steam distillation and extraction) method using pentane and diethyl ether(1:1), and their volatile flavor compounds were analyzed by GC and GC-MS. Total 51 components, including 10 hydrocarbons, 15 alcohols, 12 aldehydes, 4 ketones, 4 esters and 6 acids were identified in fresh Eucommia ulmoides harvested in July. In fresh samples harvested in September, 15 hydrocarbons, 10 alcohols, 5 aldehydes, 4 ketones, 4 esters and 3 acids were identified. In fresh Eucommia ulmoides, aldehydes(8.25ppm) were the most abundant compounds in July samples and alcohols(18.87ppm) in September ones. Seventy one components, including 21 hydrocarbons, 12 alcohols, 12 aldehydes, 9 ketones, 5 esters, 8 acids and 4 miscellaneous ones were identified in air-dried samples harvested in July. In air-dried samples harvested in September, 10 hydrocarbons, 9 alcohols, 3 aldehydes, 3 ketones, 4 esters, 4 acids and 1 miscellaneous one were identified, and the most abundant compounds in July and September samples were hydrocarbons at 5.06ppm and 15.11ppm, respectively. A total of 41 components, including 13 hydrocarbons, 9 alcohols, 5 aldehydes, 3 ketones, 6 esters and 5 acids were identified in freeze-dried samples harvested in July. Freeze-dried samples harvested in August also contained 41 components but with different types, and the ones of September 26 compounds. In freeze-dried ones, hydrocarbons were the most abundant compounds in July sample and esters in August and September samples.

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Muskelin Interacts with Multi-PDZ Domain Protein 1 (MUPP1) through the PDZ Domain (Muskelin과 multi-PDZ domain protein 1 (MUPP1) 단백질의 PDZ 도메인을 통한 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.25 no.5
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    • pp.594-600
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    • 2015
  • Protein-protein interactions have a critical role in the regulation of many cellular functions. Postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domain is one of domains that mediate protein-protein interactions. PDZ domains typically bind to the specific motif at the carboxyl (C)-terminal end of partner proteins. Multi-PDZ domain protein 1 (MUPP1), which has 13 PDZ domains, serves a scaffolding function for structure proteins and signaling proteins, but the cellular function of MUPP1 has not been fully elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and muskelin. Muskelin was recently identified as a GABAA receptor (GABAAR) α1 subunit binding protein and known to have a role in receptor endocytosis and degradation. Muskelin bound to the 3rd PDZ domain, but not to other PDZ domains of MUPP1. The C-terminal end of muskelin was essential for the interaction with MUPP1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, muskelin but not the C-terminal deleted muskelin was co-immunoprecipitated with MUPP1. In addition, MUPP1 co-localized with muskelin at the same subcellular region in cells. These findings collectively suggest that MUPP1 or its interacting proteins could modulate GABAAR trafficking and turnover through the interaction with muskelin.

Effect of NaCl on the Stability of Oncolytic Vaccinia Virus (항암 백시니아 바이러스의 안전성에 대한 염화나트륨의 효과)

  • Kim, Seong-Geun;Ran, Gui Shao;Kwon, Hyuk-Chan;Hwang, Tae-Ho
    • Journal of Life Science
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    • v.26 no.1
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    • pp.23-33
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    • 2016
  • Pexa-Vec (JX-594) is a specific cancer-targeted oncolytic and immunotherapeutic vaccinia virus. The purpose of this study was to develop methods to maximize the stability of Pexa-Vec. In short-term instability testing, viral activity was rapidly decreased both at 4℃ and at room temperature (RT), but it was completely restored after sonication followed by vortex. Long-term stability testing of Pexa-Vec in the following liquid formulations was performed: (A) 30 mM Tris/pH 7.6, (B) 30 mM Tris/pH 8.6, (C) 30 mM Tris/pH 7.6, 150 mM NaCl, 15% sucrose, (D) 30 mM Tris/pH 7.6, 15% sucrose, and (E) 30 mM Tris/pH 8.6, 15% sucrose. Viral activity decreased less than 2 log10 at 4℃, and RT was observed in 3 days in B, while viral activity was not decreased even after 4–8 weeks at 4℃ and at 1 week in RT in A, suggesting that neutral pH may be essential to maintain virus stability. The addition of 15% sucrose into A (D) significantly increased viral stability at −20℃, 4℃, or RT, and it was also observed at pH 8.6 (E). The addition of 150 mM NaCl into D (C) significantly increased viral stability in addition to the sucrose effect at 4℃ or RT. Accordingly, the viral activity in formulation C was maintained for 1.5 years at 4℃, and for 1-2 weeks in RT. In conclusion, we propose that formulation C can provide the most adequate condition for the proper storage of vaccinia oncolytic virus.

Expression and Localization of 20α-Hydroxysteroid Dehydrogenase in Immature Pig Testis (미니돼지 정소에서 20α-HSD의 발현 및 특성화)

  • Kim, Jeong-Soo;Seong, Hun-Ki;Byambaragchaa, Munkhzaya;Sim, Bo-Woong;Her, Chang-Gi;Kang, Myung-Hwa;Min, Kwan-Sik
    • Journal of Life Science
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    • v.27 no.7
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    • pp.739-745
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    • 2017
  • In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy. $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) predominantly converts progesterone into its biologically inactive form $20{\alpha}$-hydroxyprogesterone ($20{\alpha}$-OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In this study, we characterized the expression and localization of $20{\alpha}$-HSDinthe testis of MediKinetics $Micropigs^{(R)}$. The testes were collected at days 6, 9, 12, 18, and 21 after birth. The $20{\alpha}$-HSD mRNA was found to be expressed in the testis at day 6 after birth by RT-PCR. The highest level of mRNA expression in the testis was detected on day 21 after birth. However, the mRNA was not detected in the placenta after parturition. Western blot for $20{\alpha}$-HSD reveal that the specific 37-kDa band was detected in immature pig testis. However, this band was not detected in testis tissue at day 6 after birth. In the immunohistochemical analysis of the testis, $20{\alpha}$-HSD was detected in the Sertoli cells and Leydig cells. Taken together, our study shows for the first time that the $20{\alpha}$-HSD mRNA and protein are expressed in pig testis after birth. Further investigation is required to elucidate the functional mechanisms of $20{\alpha}$-HSD in pig testis after birth.

Volatile Flavor Components and Free Radical Scavenging Activity of Cnidium officinale (천궁(Cnidium officinale)의 휘발성 향기성분 및 유리기 소거활성)

  • Lee, Ji-Hye;Choi, Hyang-Sook;Chung, Mi-Sook;Lee, Mie-Soon
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.330-338
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    • 2002
  • This study was performed to develop natural spices and functional foods using Cheongung (Cnidium officinale) which is one of the Korean medicinal plants. The volatile flavor patterns of Cnidium officinale were detected by electronic nose with 6 metal oxide sensors, and the principal component analysis was carried out. The volatile flavor components of Cnidium officinale were isolated by simultaneous steam-distillation extraction with pentane and diethylether (1 : 1), and essential oils were analyzed by capillary GC and GC/MS. The free radical scavenging activity of ethanol and methanol extracts from Cnidium officinale was measured by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and compared with ${\alpha}-tocopherol$ as reference. The principal component analysis showed the difference of principal components between fresh and drying samples. Eighty-five volatile flavor components (643.64 ppm) from fresh Cnidium officinale were identified and the major components were butyl phthalide, sabinene, neocnidilide. Sixty-four volatile flavor components (218.15 ppm) from hot air dried one were identified and the major components were butyl phthalide, sabinene, 3-N-butyl phthalide. And 73 volatile flavor components (784.15 ppm) from freeze dried one were identified and the major components were butyl phthalide, sabinene, ${\beta}-selinene$. The free radical scavenging activity of methanol cold extract (500 ppm) of freeze dried Cnidium officinale was higher than other samples. And methanol and ethanol cold extracts (above 250 ppm) of freeze dried sample were higher than ${\alpha}-tocopherol$ $25\;{\mu}M$ (22.34%).