• Title/Summary/Keyword: GV20

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Simultaneous Detection of Seven Phosphoproteins in a Single Lysate Sample during Oocyte Maturation Process (난자성숙 과정의 단일 시료에서 일곱 가지 인산화 단백질의 동시 분석 방법)

  • Yoon, Se-Jin;Kim, Yun-Sun;Kim, Kyeoung-Hwa;Yoon, Tae-Ki;Lee, Woo-Sik;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.3
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    • pp.187-197
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    • 2009
  • Objective: Phosphorylation and dephosphorylation of proteins are important in regulating cellular signaling pathways. Bead-based multiplex phosphorylation assay was conducted to detect the phosphorylation of seven proteins to maximize the information obtained from a single lysate of stage-specific mouse oocytes at a time. Methods: Cumulus-oocyte complexes (COCs) were cultured for 2 h, 8 h, and 16 h, respectively to address phosphorylation status of seven target proteins during oocyte maturation process. We analyzed the changes in phosphorylation at germinal vesicle (GV, 0 h), germinal vesicle breakdown (GVBD, 2 h), metaphase I (MI, 8 h), and metaphase II (MII, 16 h in vitro or in vivo) mouse oocytes by using Bio-Plex phosphoprotein assay system. We chose seven target proteins, namely, three mitogen-activated protein kinases (MAPKs), ERK1/2, JNK, and p38 MAPK, and other 4 well known signaling molecules, Akt, GSK-$3{\alpha}/{\beta}$, $I{\kappa}B{\alpha}$, and STAT3 to measure their phosphorylation status. Western blot analysis and kinase inhibitor treatment for ERK1/2, JNK, and Akt during in vitro maturation of oocytes were conducted for the confirmation. Results: Phosphorylation of ERK1/2, JNK, p38 MAPK and STAT3 was increased over 3 folds up to 20 folds, while phosphorylation of the other three signal molecules, Akt, GSK-$3{\alpha}/{\beta}$, and $I{\kapa}B{\alpha}$ was less than 3 folds. All of these results except for Akt were statistically significant (p<0.05). Conclusion: This is the first report on the new and valuable method measuring many phosphoproteins simultaneously in one minute sample such as oocyte lysates. All of the three MAPKs, ERK1/2, JNK, and p38 MAPK are involved in the process of mouse oocyte maturation. In addition, STAT3 might be important regulator of oocyte maturation, while Akt phosphorylation at Serine 473 may not be involved in the regulation of oocyte maturation.

Land-Cover Change Detection of Western DMZ and Vicinity using Spectral Mixture Analysis of Landsat Imagery (선형분광혼합화소분석을 이용한 서부지역 DMZ의 토지피복 변화 탐지)

  • Kim, Sang-Wook
    • Journal of the Korean Association of Geographic Information Studies
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    • v.9 no.1
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    • pp.158-167
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    • 2006
  • The object of this study is to detect of land-cover change in western DMZ and vicinity. This was performed as a basic study to construct a decision support system for the conservation or a sustainable development of the DMZ and Vicinity near future. DMZ is an is 4km wide and 250km long and it's one of the most highly fortified boundaries in the world and also a unique thin green line. Environmentalists want to declare the DMZ as a natural reserve and a biodiversity zone, but nowadays through the strengthening of the inter-Korean economic cooperation, some developers are trying to construct a new-town or an industrial complex inside of the DMZ. This study investigates the current environmental conditions, especially deforestation of the western DMZ adopting remote sensing and GIS techniques. The Land-covers were identified through the linear spectvral mixture analysis(LSMA) which was used to handle the spectral mixture problem of low spatial resolution imagery of Landsat TM and ETM+ imagery. To analyze quantitative and spatial change of vegetation-cover in western DMZ, GIS overlay method was used. In LSMA, to develop high-quality fraction images, three endmembers of green vegetation(GV), soil, water were driven from pure features in the imagery. Through 15 years, from 1987 to 2002, forest of western DMZ and vicinity was devastated and changed to urban, farmland or barren land. Northern part of western DMZ and vicinity was more deforested than that of southern part. ($52.37km^2$ of North Korean forest and $39.04km^2$ of South Korean were change to other land-covers.) In case of North Korean part, forest changed to barren land and farmland and in South Korean part, forest changed to farmland and urban area. Especially, In North Korean part of DMZ and vicinity, $56.15km^2$ of farmland changed to barren land through 15 years, which showed the failure of the 'Darakbat' (terrace filed) project which is one of food increase projects in North Korea.

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Interspecies Nuclear Transfer using Bovine Oocytes Cytoplasm and Somatic Cell Nuclei from Bovine, Porcine, Mouse and Human (소, 돼지, 생쥐, 사람의 체세포와 소 난자를 이용한 이종간 핵 이식)

  • 박세영;김은영;이영재;윤지연;길광수;김선균;이창현;정길생;박세필
    • Korean Journal of Animal Reproduction
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    • v.26 no.3
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    • pp.235-243
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    • 2002
  • This study was designed to examine the ability of the bovine (MII) oocytes cytoplasm to support several mitotic cell cycles under the direction of differentiated somatic cell nuclei of bovine, porcine, mouse and human. Bovine GV oocytes were matured in TCM-199 supplemented with 10% FBS. At 20h after IVM, recipient oocytes were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst and their 1st polar body (PB) and MII plate were removed by enucleation micropipette under UV filter. Ear skin samples were obtained by biopsy from an adult bovine, porcine, mouse and human and cultured in 10% FBS added DMEM. Individual fibroblast was anlaysed chromosome number to confirm the specificity of species. Nuclear transferred (NT) units were produced by electrofusion of enucleated bovine oocytes with individual fibroblast. The reconstructed embryos were activated in 5 $\mu$M ionomycin for 5 min followed by 1.9 mM 6-dimethylaminopurine (DMAP) in CR1aa for 3 h. And cleaved NT embryos were cultured in CR1aa medium containing 10% FBS on monolayer of bovine cumulus cell for 8 days. Also NT embryo of 4~8 cell stage was analysed chromosome number to confirm the origin of nuclear transferred somatic cell. The rates of fusion between bovine recipient oocytes and bovine, porcine, mouse and human somatic cells were 70.2%, 70.2%, 72.4% and 63.0%, respectively. Also, their cleavage rates were 60.6%, 63.7%, 54.1% and 62.7%, respectively, there were no differences among them. in vitro development rates into morula and blastocyst were 17.5% and 4.3% in NT embryos from bovine and human fibroblasts, respectively. But NT embryos from porcine and mouse fibroblasts were blocked at 16~32-cell stage. The chromosome number in NT embryos from individual fibroblast was the same as chromosome number of individual species. These results show that bovine MII oocytes cytoplasm has the ability to support several mitotic cell cycles directed by newly introduced nuclear DNA.

Short Tenn Reactions to Acupuncture Treatment and Adverse Events Following Acupuncture in Korea a Cross-sectional Survey of Patient Reports (침치료 직 후 자가 설문지를 이용한 침반응(針感)과 부작용에 대한 단면적 연구)

  • Park, Seong-Uk;Jung, Woo-Sang;Moon, Sang-Kwan;Ko, Chang-Nam;Cho, Ki-Ho;Kim, Young-Suk;Bae, Hyung-Sup;Park, Jung-Mi
    • The Journal of Korean Medicine
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    • v.28 no.2 s.70
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    • pp.66-79
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    • 2007
  • Objectives : To explore the type and frequency of short term reactions, de Qi associated with acupuncture treatment and to determine the incidence of adverse events following acupuncture in Korea. Subjects and methods : This study is a retrospective and cross-sectional survey of patient reports. 1095 subjects, 585 of out-patients of the Oriental Medicine of Stroke & Neurological Disorders Center, East-West NEO Medical Center of Kyunghee University and 510 of out-patients of the Department of Cardiovascular & Neurologic Diseases (Stoke Center), Hospital of Oriental Medicine, Kyunghee Medical Center, from June through November of 2006, who had acupuncture, gave informed consent and completed one survey form. On this form, patients were asked to report short term acupuncture reactions, de Qi, patient satisfaction measurement (using VAS), and adverse events relating to acupuncture treatment. The acupuncturists of this study are Korean Medicine Doctors (KMD) who had worked as practitioners for 3-30 years or more. Results : The average age of the 1095 subjects was 58 years old. Positive short term acupuncture reactions after treatment were reported by 878 (80.2%), negative short term acupuncture reactions by 75 (6.8%) and no reactions were reported by 142 (13.0%). The most common positive short term acupuncture reactions were feeling 'relaxed', 472 (43.1%), followed by feeling 'less pain' 90 (8.2%), 'energized' 16 ( 1.5%), 'tingling' 16 (1.5%), 'heat feeling or Cold feeling' 10 (0.9%), and others 274 (25.0%), respectively. Negative short term acupuncture reactions were feeling 'pain' 37 (3.4%), tiredness 24 (2.2%), dizziness 9 (0.8%), and others 5 (0.2%), respectively. Traditionally described needling sensations of de Qi refer to a patient's response to distention, pulling, soreness, heaviness, numbness. 39.7% of subjects reported de Qi during needling, experiencing 'distention' 333 (30.4%), 'soreness' 52 (4.7%), 'pulling' 22 (2.0%), 'heaviness' 18 (1.6%), and 'numbness' 10 (0.9%) respectively. Positive short term acupuncture reactions and de Qi rate were the highest in the less than 40 years group (83/96 86.5%, 50/96 52.1%). No acupuncture reactions were highly seen in the over 70 years old group (31/187, 16.6%). Patient satisfaction level using VAS was a comparatively high $72.9{\pm}19.9$. Adverse events were only bleeding in 92 (8.4%) of the total subjects. High sensitive acupoints were 95 points as GV26 (54 times), LI4 (54 times), ST36 (53 times), GB20 (37 times), HT8(34 times), LV3 (29 times), SI3 (29 times), and LI11 (27 times) in order. Main impressions were stroke patients 430 (16.9%), headache 185 (16.9%), hypertension 97 (8.9%), and dizziness 85 (7.8%). Conclusions : Although 8 different Korean Medicine Doctors participated in this research, we obtained similar results from each. There were no significantly different results between the two hospitals. Short term acupuncture reactions and de Qi were most related to age. Except for bleeding there were no adverse events relating to acupuncture treatment in this study. We consider acupuncture treatment as very safe depending on practitioners. Positive short term acupuncture reactions after treatment were 12 times higher than negative short term acupuncture reactions. Subjects were comparatively satisfied with acupuncture treatment.

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The Effect of Acupuncture Treatment on the Heart Rate Variability of Chronic Headache Patients (만성두통환자에 대한 침치료가 심박변이도에 미치는 영향)

  • Jung, In-tae;Lee, Sang-hoon;Kim, Su-young;Cha, Nam-hyun;Kim, Keon-sik;Lee, Doo-ik;Lee, Jae-Dong;Lim, Sabina;Lee, Yun-ho;Choi, Do-young
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.105-112
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    • 2005
  • Obiective : The purpose of this study was to assess the effect of acupuncture treatment for chronic headache patients using power spectrum analysis of the heart rate variability(HRV). Methods : 15 clinical experiment participants were gathered and through a questionnaire patients who experienced headache for more than 4 hours a day and more than IS days per month were qualified as Chronic Headache patients. Treatment was afplied 2 times a weeks for 8 weeks. The acupoints, GV2O, HN23, ST8, HN46, TEl7, GB2O, LI2O, LI11, LI14, ST36, and LR3 were stimulated for 20 minutes. The effects of acupuncture treatment were analyzed using power spectrum analysis of the HRV. HRV was recorded before and after acupuncture treatment. Results : HRV before and after treatment was compared after 8 weeks of acupuncture treatment. Increase in mean values of SDNN and RMSSD were observed but the increases were not statistically significant. Increase in mean values of TP, LF and HF were observed but, the increase was significant(p<0.05) only in TP. Conclusions : The results suggest that acupuncture treatment on chronic headache patients can increase the activity of autonomic nervous system. Further use of HRV for quantitative analysis of acupuncture treatment on autonomic nervous system related symptoms is suggested.

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