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Sensitization of the Apoptotic Effect of ${\gamma}$-Irradiation in Genistein-pretreated CaSki Cervical Cancer Cells

  • Shin, Jang-In;Shim, Jung-Hyun;Kim, Ki-Hong;Choi, Hee-Sook;Kim, Jae-Wha;Lee, Hee-Gu;Kim, Bo-Yeon;Park, Sue-Nie;Park, Ok-Jin;Yoon, Do-Young
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.523-531
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    • 2008
  • Radiotherapy is currently applied in the treatment of human cancers. We studied whether genistein would enhance the radiosensitivity and explored its precise molecular mechanism in cervical cancer cells. After co-treatment with genistein and irradiation, the viability, cell cycle analysis, and apoptosis signaling cascades were elucidated in CaSki cells. The viability was decreased by co-treatment with genistein and irradiation compared with irradiation treatment alone. Treatment with only ${\gamma}$-irradiation led to cell cycle arrest at the $G_1$ phase. On the other hand, co-treatment with genistein and ${\gamma}$-irradiation caused a decrease in the $G_1$ phase and a concomitant increase up to 56% in the number of $G_2$ phase. In addition, co-treatment increased the expression of p53 and p21, and Cdc2-tyr-15-p, supporting the occurrence of $G_2/M$ arrest. In general, apoptosis signaling cascades were activated by the following events: release of cytochrome c, upregulation of Bax, down regulation of Bcl-2, and activation of caspase-3 and -8 in the treatment of genistein and irradiation. Apparently, co-treatment downregulated the transcripts of E6*I, E6*II, and E7. Genistein also stimulated irradiation-induced intracellular reactive oxygene, species (ROS) production, and co-treatment-induced apoptosis was inhibited by the antioxidant N-acetylcysteine, suggesting that apoptosis has occurred through the increase in ROS by genistein and ${\gamma}$-irradiation in cervical cancer cells. Gamma-irradiation increased cyclooxygenase-1 (COX-2) expression, whereas the combination with genistein and ${\gamma}$-irradiation almost completely prevented irradiation-induced COX-2 expression and $PGE_2$ production. Co-treatment with genistein and ${\gamma}$-irradiation inhibited proliferation through $G_2/M$ arrest and induced apoptosis via ROS modulation in the CaSki cancer cells.

Effect on Gene Expression Profile of Rat Hippocampus Caused by Administration of Memory Enhancing Herbal Extract (육미지황탕가미방이 흰쥐의 기억능력과 중추신경계 유전자 발현에 미치는 영향)

  • Choi So Eop;Bae Hyun Su;Shin Min Kyu;Hong Moo Chang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.5
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    • pp.1025-1034
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    • 2002
  • The herbal extract (YMT_02) is a modified herbal extracts from Yukmijihwang-tang (YMJ) to promote memory-enhancing. The YMJ extracts has been widely used as an anti-aging herbal medicine for hundred years in Asian countries. The purpose of this study is to; 1) quantitatively evaluate the memory-enhancing effect of YMT_02 by behavior task, 2) identify candidate genes responsible for enhancing memory by cDNA microarray and 3) assess the anti-oxidant effect of YMT_02 on PC12 cell. Memory retention abilities are addressed by passive avoidance task with Sprague-Dawley (SD) male rat. Before the training session, the rats are subdivided into four groups and administrated with YMT_02, Ginkgo biloba, Soya lecithin and normal saline for 10 days. The retention test was performed. 24 hours after the training session. The retention time of the YMT_02 group was significantly (p<0.05) delayed (~100%), whereas Ginkgo biloba and Soya lecithin treatment delayed 20% and 10% respectively. The hippocampi of YMT_02 and control group were dissected and mANA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Incyte rat GEMTM 2 cDNA microarray. The microarray results show that prealbumin(transthyretin), phosphotidylethanolamine N-methyltransferase, and PEP-19 are expressed abundantly in the YMT_02 treated group. Especially, PEP-19 is a neuron-specific protein, which inhibits apoptotic processes in neuronal cell. On the other hand, transcripts of RAB15, glutamate receptor subunit 2 and CDK108 are abundant in control group. Besides, neuronal genes involved in neuronal death or neurodegeneration such as neuronal-pentraxin and spectrin are abundantly expressed in control group. Additionally, the YMT_02 shows an anti oxidative effect in the PC12 cell. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the memory-enhancing effect of herbal extracts YMT_02, for example, anti-apoptotic, anti-oxidative, and neuroprotective effects.

Criteria for Evaluating Scientific Models Used by Pre-service Elementary Teachers (예비 초등 교사들의 과학 모델 평가 기준)

  • Oh, Phil Seok;Lee, Jung Sook
    • Journal of The Korean Association For Science Education
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    • v.34 no.2
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    • pp.135-146
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    • 2014
  • The purpose of this study is to explore evaluation criteria that pre-service elementary teachers employ as they evaluate and select models to explain electric circuits. Thirty junior students in a university of education have participated in the study as a part of the science education course in which they were enrolled. The lessons for the participants have been organized as a cyclic sequence of different modeling pedagogies including the expressive, experimental, and evaluative modeling. The pre-service teachers have been given five electric circuits in order and asked to create models and further develop them through peer discussion. Their modeling activities have been video- or audio-recorded, and the recordings and their transcripts have been analyzed using a framework of model evaluation criteria. It reveals that the types and frequencies of evaluation criteria used are different between situations of model development and model selection. While empirical and theoretical criteria have been used dominantly in both situations, more various criteria have been employed in the situation where the pre-service teachers selected one model among alternatives. Implications for science education and science education research have been suggested.

Molecular Cloning, Tissue Distribution and Segmental Ontogenetic Regulation of b0,+ Amino Acid Transporter in Lantang Pigs

  • Zhi, Ai-Min;Feng, Ding-Yuan;Zhou, Xiang-Yan;Zou, Shi-Geng;Huang, Zhi-Yi;Zuo, Jian-Jun;Ye, Hui;Zhang, Chang-Ming;Dong, Ze-Min;Liu, Zhun
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.8
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    • pp.1134-1142
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    • 2008
  • Cationic amino acid transporter $b^{0,+}AT$ (HGMW-approved gene symbol SLC7A9, solute carrier family 7, member 9) plays a crucial role in amino acid nutrition. In the present study, we describe the cloning and sequencing of porcine $b^{0,+}AT$. Based on the sequence of porcine $b^{0,+}AT$ deposited in the NCBI (National Center for Biotechnological Information), we identified a putative porcine homologue. Using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine $b^{0,+}AT$ was isolated. The porcine $b^{0,+}AT$ cDNA was 1,680 bp long, encoding a 487 amino acid trans-membrane protein. The predicted amino acid sequence was found to have 88.9% and 87.1% identity with human and mouse $b^{0,+}AT$, respectively. Real-time RT-PCR indicated porcine $b^{0,+}AT$ transcripts expressed in heart, kidney, muscle and small intestine. The small intestine had the highest $b^{0,+}AT$ mRNA abundance while the muscle had the lowest (p<0.05). Along the longitudinal axis, the ileum had the highest $b^{0,+}AT$ mRNA abundance while the colon had the lowest (p<0.05). The $b^{0,+}AT$ mRNA level was highest on day 7 and 90 in the duodenum (p<0.05). It increased from day 1 to day 26 in the jejunum (p>0.05) and had the highest abundance on day 60 (p<0.05). There was, however, no difference between day 1, 7, 26, 30, 90 and 150 (p>0.05). The strongest $b^{0,+}AT$ expression appeared on day 7 in the ileum before weaning, and then decreased till day 30 but rose gradually again from day 60 to 150 (p<0.05).

Molecular Cloning and Expression of the Novel Attacin-Like Antibacterial Protein Gene Isolated from the Bombyx mori (집누에로부터 새로운 attactin 유산 항세균성 펩타이드 유전자의 분리 및 발현)

  • 윤은영;김상현;강석우;진병래;김근영;김호락;한명세;강석권
    • Korean journal of applied entomology
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    • v.36 no.4
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    • pp.331-340
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    • 1997
  • Hyalophora cecropia attacin-like antibacterial gene was isolated from Bombyx mori induced with nonpathogenic bacteria. It was expressed in Spodopfera frugiperda 9 (Sf9 cells using baculovirus expression vector system (BEVS), and examined its antibacterial activity. With a cDNA library constructed from fifthinstar B. mori injected with Escherichia coli(4 X IOhcellsllarva), differential screening was performed using naive and induced mRNA probes. BmInc6 clone was screened by partial nucleotide sequence and GenBank database analysis. A complete nucleotide sequence of Bmlnc6 cDNA was determined (GenBank, AF005384). Its insert size was 852 bp and had open reading frame that started translation at position 35 and stopped at 679. And its putative polyadenylational signal existed at 812 bp. The number of amino acid deduced from Bmlnc6 cDNA was 214 and hydropathy analysis showed that this peptide was hydrophilic. This peptide deduced by BmInc6 was named nuecin. When the nuecin gene was expressed in Sf9 cells using BEVS, about 950 bp of the transcripts was detected. In addition, SDS-PAGE analysis showed that the molecular weights of intracellular expressed protein and the mature protein secreted to culture media were approximately 23 and 20 kDa, respectively. The antibacterial activity of nuecin against E. coli and Bacillus subtilis was significantly high, demonstrating that nuecin had a wider antibacterial spectrum with gram negative and positive bacteria than attacin.

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A Story of Practice of Mutual Respect Rights between a Mother and two Sons (어머니와 두 아들의 상호존중 권리실천 이야기)

  • Suh, Young Sook;Kim, Ji Eun;Song, Eun Kyung
    • Korean Journal of Childcare and Education
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    • v.9 no.6
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    • pp.439-460
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    • 2013
  • The purpose of this study was to examine a mother's and her two son's 'practice of mutual respect rights' experiences, and to find a solution to inspire the practice of realizing rights of mutual respect within a family. From September 2012 to March 2013, qualitative research methods such as transcripts, observations, in-depth interviews, and analysis of records were applied to a mother and her two sons. As a result, the mother's experiences showed that she and her two sons had enough mutual respect, and the regression of the first child was a natural and positive part in that process. Also, conflicts between protection rights and participation rights were integrated into a practice of mutual respect, and the mother had an opportunity to think about positive discipline. With this practice, the mother and her two sons were able to reconcile their relationships and have deep respect for one another, and family members felt psychologically safe through this practice of rights. In addition, the practice of rights was transferred to other family members, and the mother came to have courage about being an adequate parent. The results of this study revealed that the following research could be used to propose concrete contents and to plan how to recognize and access the practice of rights among family members.

Current Barriers of Obesity Management of Children Using Community Child Care Centers and Potential Possibility of Utilizing Mobile Phones: A Qualitative Study for Children and Caregivers (지역아동센터 이용 어린이의 비만관리의 한계점과 모바일폰의 잠재적인 활용 가능성: 어린이와 보호자 대상의 질적 연구)

  • Lee, Bo Young;Park, Mi-Young;Kim, Kirang;Shim, Jea Eun;Hwang, Ji-Yun
    • Korean Journal of Community Nutrition
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    • v.25 no.3
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    • pp.189-203
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    • 2020
  • Objectives: This study was performed to identify the current barriers of obesity management for children using Community Child Care Centers and their caregivers (parents and teachers working in the Centers). Further, this study explored the possibility of utilizing a mobile phone application for tailored obesity prevention and management programs to overcome the current difficulties associated with children's obesity management. Methods: The qualitative data were collected through in-depth interviews with 20 obese and overweight children or children who wanted to participate in this study using Community Child Care Centers, 12 teachers working at the Centers, and a focus group interview with five parents of children using the Centers. Data were analyzed with a thematic approach categorizing themes and sub-themes based on the transcripts. Results: The current barriers of obesity management of obese and overweight children using Community Child Care Centers were lack of self-directed motivation regarding obesity management (chronic obesity-induced lifestyles and reduced self-confidence due to stigma) and lack of support from households and Community Child Care Centers (latchkey child, inconsistency in dietary guidance between the Center and household, repetitive pressure to eat, and absence of regular nutrition education). Mobile phone applications may have potential to overcome the current barriers by providing handy and interesting obesity management based on visual media (real-time tracking of lifestyles using behavior records and social support using gamification), environmental support (supplementation of parental care and network-based education between the Community Child Care Center and household), and individualized intervention (encouragement of tailored and gradual changes in eating habits and tailored goal setting). It is predicted that the real-time mobile phone program will provide information for improving nutritional knowledge and behavioral skills as well as lead to sustainable children's coping strategies regarding obesity management. In addition, it is expected that environmental factors may be improved by network-based education between the Community Child Care Centers and households using the characteristics of mobile phones, which are free from space and time constraints. Conclusions: The tailored education program for children using Community Child Care Centers based on mobile phones may prevent and reduce childhood obesity by overcoming the current barriers of obesity management for children, providing environmental and individualized support to promote healthy lifestyles and quality of life in the future.

Characterization of Gibberellic Acid-Stimulated Arabidopsis (GASA) gene to drought stress response in Poplar (Populus alba × P. glandulosa) (현사시나무 Gibberellic Acid-Stimulated Arabidopsis (GASA) 유전자의 발현 특성 및 건조 스트레스 내성 구명)

  • Choi, Hyunmo;Bae, Eun-Kyung;Choi, Young-Im;Yoon, Seo-Kyung;Lee, Hyoshin
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.61-68
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    • 2017
  • Gibberellic Acid-Stimulated Arabidopsis (GASA) genes are involved in plant hormone signaling, cell division and elongation, as well as in responses to stress conditions in plants. In this study, we isolated a GASA gene from hybrid poplar (Populus alba ${\times}$ P. glandulosa) and analyzed its physiological phenotype and molecular functions in poplar. PagGASA cDNA encodes a putative protein composed of 95 amino acids containing an N-terminal signal peptide and a conservative cysteine-rich C-terminal domain. Southern blot analysis revealed that one or two copies of the PagGASA are present in the poplar genome. The PagGASA transcripts were highly detected in flowers and roots. Moreover, the expression of PagGASA was induced by growth hormone (gibberellic acid) and stress hormones (abscisic acid, jasmonic acid, and salicylic acid). By using transgenic analysis, we showed that the upregulation of PagGASA in poplar provides high tolerance to drought stress. Therefore, our results suggest that PagGASA plays an important role in drought stress tolerance via stress-related plant hormone signaling in poplar.

Recognition of Transmembrane Protein 39A as a Tumor-Specific Marker in Brain Tumor

  • Park, Jisoo;Lee, Hyunji;Tran, Quangdon;Mun, Kisun;Kim, Dohoon;Hong, Youngeun;Kwon, So Hee;Brazil, Derek;Park, Jongsun;Kim, Seon-Hwan
    • Toxicological Research
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    • v.33 no.1
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    • pp.63-69
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    • 2017
  • Transmembrane protein 39A (TMEM39A) belongs to the TMEM39 family. TMEM39A gene is a susceptibility locus for multiple sclerosis. In addition, TMEM39A seems to be implicated in systemic lupus erythematosus. However, any possible involvement of TMEM39A in cancer remains largely unknown. In the present report, we provide evidence that TMEM39A may play a role in brain tumors. Western blotting using an anti-TMEM39A antibody indicated that TMEM39A was overexpressed in glioblastoma cell lines, including U87-MG and U251-MG. Deep-sequencing transcriptomic profiling of U87-MG and U251-MG cells revealed that TMEM39A transcripts were upregulated in such cells compared with those of the cerebral cortex. Confocal microscopic analysis of U251-MG cells stained with anti-TMEM39A antibody showed that TMEM39A was located in dot-like structures lying close to the nucleus. TMEM39A probably located to mitochondria or to endosomes. Immunohistochemical analysis of glioma tissue specimens indicated that TMEM39A was markedly upregulated in such samples. Bioinformatic analysis of the Rembrandt knowledge base also supported upregulation of TMEM39A mRNA levels in glioma patients. Together, the results afford strong evidence that TMEM39A is upregulated in glioma cell lines and glioma tissue specimens. Therefore, TMEM39A may serve as a novel diagnostic marker of, and a therapeutic target for, gliomas and other cancers.

Effects of Glucose and IGF-I on Expression of Glucose Transporter 1 (Glut1) and Development of Preimplantation Mouse Embryo (생쥐의 착상전 배아의 발생과 Glucose Transporter 1 (Glut1) 발현에 대한 포도당과 IGF-I의 영향)

  • 전한식;계명찬;김종월;강춘빈;김문규
    • Development and Reproduction
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    • v.2 no.2
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    • pp.205-212
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    • 1998
  • A sodium-independent facilitative glucose transporter 1 (Glut1) is a major route by which glucose can be transported across the plasma membrane of mouse embryo. Although it has been known that insulin-like growth factor-I (IGF-I) promotes glucose transport into the mouse embryo, whether IGF-I directly regulates transcription of Glut1 has been uncovered in mouse preimplantation embryo. This study was aimed to elucidate the role of glucose and IGF-I in development and Glut1 expression in preimplantation mouse embryo. Two-cell embryos developed in blastocyst regardless of the glucose in the presence of pyruvate. IGF-I significantly increased the number of blastomeres in the mid-blastula. Deprivation of glucose did not affect the amount of Glut1 transcripts in morula cultured from 2-cell embryo. IGF-I potentiated Glut1 expression in morula cultured from 2-cell embryo even in the absence of glucose. Taken together, it is concluded that depletion of glucose does not promote Glut1 expression the in morula cultured form 2-cell embryo, and that increment of Glut1 expression possibly mediates embryotropic effect of IGF-I on preimplantation mouse embryo.

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