The shortage of human organs for transplantation has induced the research on the possibility of using animal as porcine. However, pig to human transplantation as known as xeno-transplantation has major problem as immunorejection. Recently, the solutions of pig to human xenotransplantation are commonly mentioned as having a genetically modification which include alpha 1, 3 galatosyl transferase knockout (GTKO) and immune-suppressing gene transgenic model. Unfortunately, the expression level of transgenic gene is very low activity. Therefore, development of gene overexpression system is the most urgent issue. Also, the tissue specific overexpression system is very important. Because most blood vessels are endothelial cells, establishment of the endothelial-specific promoter is attractive candidates for the introduction of suppressing immunorejection. In this study, we focus the ICAM2 promoter which has endothelial-specific regulatory region. To detect the regulatory region of ICAM2 promoter, we cloned 3.7 kb size mini-pig ICAM2 promoter. We conduct serial deletion of 5' flanking region of mini-pig ICAM2 promoter then selected promoter size as 1 kb, 1.5 kb, 2 kb, 2.5 kb, and 3 kb. To analyze promoter activity, luciferase assay system was conducted among these vectors and compare endothelial activity with epithelial cells. The reporter gene assay revealed that ICAM2 promoter has critical activity in endothelial cells (CPAE) and 1 kb size of ICAM2 promoter activity was significantly increased. Taken together, our studies suggest that mini-pig ICMA2 promoter is endothelial cell specific overexpression promoter and among above all size of promoters, 1 kb size promoter is optimal candidate to overcome the vascular immunorejection in pig to human xenotransplantation.
Objective: The aim of this study is to elucidate the effects of transforming growth factor-${\beta}$ (TGF-${\beta}$)1 and L-ascorbic acid on proteoglycan synthesis, and the relationship between Sox9, proteoglycan, and TGF-${\beta}1$ in intervertebral disc cells. Methods: Human intervertebral disc tissue was sequentially digested to 0.2% pronase and 0.025% collagenase in DMEM/F-12 media and extracted cells were cultured in $37^{\circ}C$, 5% $CO_2$ incubator. When intervertebral disc cells were cultured with TGF-${\beta}1$ or L-ascorbic acid, the production level of sulfated glycosaminoglycan (sGAG) was estimated by dimethyl methyleneblue (DMMB) assay. The changes of Sox9 mRNA and protein levels via TGF-${\beta}1$ were detected by RT-PCR and Western blot analysis in each. Results: The amount of sGAG was increased with the lapse of time during incubation, and sGAG content of pellet cultured cells was much larger than monolayer culture. When primary cultured intervertebral disc cells in monolayer and pellet cultures were treated by TGF-${\beta}1$ 20 ng, sGAG content of experimental group was increased significantly compared to control group in both cultures. L-Ascorbic acid of serial concentrations (50-300 ug/ml) increased sGAG content of mono layer cultured intervertebral disc cells significantly in statistics. The co-treatment of TGF-${\beta}1$ and L-ascorbic acid increased more sGAG production than respective treatment. After treating with TGF-${\beta}1$, Sox9 mRNA and protein expression rates were significantly increased in disc cells compared with the control group. Conclusion: This study suggests that TGF-${\beta}1$ would increase sulfated glycosaminoglycan (sGAG) and other proteoglycans such as versican by elevating Sox9 mRNA and protein expressions in order.
Numerous methods can be employed to teach students and farmers on soil science and fertilizer. The cartoon is one of the most effective teaching aids both for instructors and students. Instructor becomes a director of the cartoon to organize the framework of story based on the chapter of lecture. Director should adapt the regular sentence on soil science into the short and concise scenario suitable for the visual expression of the cartoon, followed by the selection of the scene for drawing. Next is to cast the instructor for the role of actor or sometimes supporter. After casting, the director starts to draw each cut of the cartoon in sequence. I published the cartoons in weekly serials in the farmer's newspaper for three years. Each cartoon was composed of eighteen cuts and formatted with the introductory, main and conclusive issues. Cartoon started with dialogues between the cast on subject to stimulate the interest by farmers, inserted the tables and graphs for the scientific knowledge and summarized the chapter. Major contents covered in the cartoons were glossary, nutrient uptake, pH, salt accumulation, moisture, pollution, chemical fertilizer, compost and other general aspects of soils. Cartoons had been very popular serial of the newspaper and are archived in the web for reviewing.
Nitric oxide(NO) has been known to play important roles in numerous physiologic processes including neurotransmission, vasorelaxation, and cellular apoptosis. Using a mouse cDNA gene chip, we examined expression patterns and time course of NO-dependent genes in mouse macrophage RAW264.7 cells. Genes shown to be upregulated more than two fold or at least at two serial time points were further selected and validated by RT-PCR. Finally, 81 selected genes were classified by function as signaling, apoptosis, inflammation, transcription, translation, ionic homeostasis and metabolism. Among those, genes related with signaling, apoptosis and inflammation, such as guanylate cyclase 1, soluble, alpha3(Gucy1a3); protein kinase C, alpha($Pkc{\alpha}$); lymphocyte protein tyrosine kinase(Lck); BCL2/adenovirus E1B 19 kDa-interacting protein(Bnip3); apoptotic protease activating factor 1(Apaf1); X-linked inhibitor of apoptosis(Xiap); cyclin G1(Ccng1); chemokine(C-C motif) ligand 4(Ccl4); B cell translocation gene 2, anti-proliferative(Btg2); lysozyme 2(Lyz2); secreted phosphoprotein 1(Spp1); heme oxygenase(decycling) 1(Hmox1); CD14 antigen(Cd14); and granulin(Grn) may play important roles in NO-dependent responses in murine macrophages.
In the web or not, all academic information have the creator which produces that information. The creator can be individual, organization, institution, or country. Most information consist of the title, author and content. The article among academic information is described by title, author, keywords, abstract, publisher, ISSN(International Standard Serial Number) and etc., and the patent information is consisted some metadata such as invention title, applicant, inventors, agents, application number, claim items etc. Most web-based academic information services provide search functions to user by processing and handling these metadata, and the search function using the author field is important. In this paper, we propose an effective indexing management for person name search, and search techniques using boosting factor and near operation based on phrase search to improve precision rate of search result. And we describe person name retrieval result with another expression name, co-authors and persons in same research field. The approach presented in this paper provides accurate data and additional search results to user efficiently.
Bang, Minji;Gonzales, Edson Luck;Shin, Chan Young;Kwon, Kyoung Ja
Biomolecules & Therapeutics
/
v.29
no.2
/
pp.144-153
/
2021
Astrocytes play various important roles such as maintaining brain homeostasis, supporting neurons, and secreting inflammatory mediators to protect the brain cells. In aged subjects, astrocytes show diversely changed phenotypes and dysfunctions. But, the study of aged astrocytes or astrocytes from aged subjects is not yet sufficient to provide a comprehensive understanding of their important processes in the regulation of brain function. In this study, we induced an in vitro aged astrocyte model through late passage cultivation of rat primary cultured astrocytes. Astrocytes were cultured until passage 7 (P7) as late passage astrocytes and compared with passage 1 (P1) astrocytes as early passage astrocytes to confirm the differences in phenotypes and the effects of serial passage. In this study, we confirmed the morphological, molecular, and functional changes of late passage astrocytes showing aging phenotypes through SA-β-gal staining and measurement of nuclear size. We also observed a reduced expression of inflammatory mediators including IL-1β, IL-6, TNFα, iNOS, and COX2, as well as dysregulation of wound-healing, phagocytosis, and mitochondrial functions such as mitochondrial membrane potential and mitochondrial oxygen consumption rate. Culture-conditioned media obtained from P1 astrocytes promoted neurite outgrowth in immature primary cultures of rat cortices, which is significantly reduced when we treated the immature neurons with the culture media obtained from P7 astrocytes. These results suggest that late passage astrocytes show senescent astrocyte phenotypes with functional defects, which makes it a suitable model for the study of the role of astrocyte senescence on the modulation of normal and pathological brain aging.
Kim, Yun-Hui;Lee, Dong-Soo;Kang, Joo-Hyun;Lee, Yong-Jin;Chung, June-Key;Lee, Myung-Chul
The Korean Journal of Nuclear Medicine
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v.38
no.1
/
pp.99-108
/
2004
Purpose: The ability to noninvasively track the migration of neural progenitor cells would have significant clinical and research implications. We generated stably transfected F3 human neural progenitor cells with human sodium/iodide symporter (hNIS) for noninvasively tracking F3. In this study, the expression patterns of hNIS gene in F3-NIS were examined according to the cultured time and the epigenetic modulation. Materials and Methods: F3 human neural stem cells had been obtained from Dr. Seung U. Kim (Ajou University, Suwon, Korea). hNIS and hygromycin resistance gene were linked with IRES (Internal Ribosome Entry Site) under control of CMV promoter. This construct was transfected to F3 with Liposome. To investigate the restoration of hNIS gene expression in F3-NIS, cells were treated with demethylating agent (5-Azacytidine) and Histone deacetylase inhibitor (Trichostatin A: TSA). The expression of hNIS was measured by I-125 uptake assay and RT-PCR analysis. Results: The iodide uptake of the F3-NIS was higher 12.86 times than F3 cell line. According to the cell passage number, hNIS expression in F3-NIS gradually diminished. After treatment of 5-Azacytidine and TSA with serial doses (up to $20{\mu}M$, up to 62.5nM, respectively) for 24 hours, I-125 uptake and mRNA of hNIS in F3-NIS were increased. Conclusion: These results suggest that hNIS transfected F3 might undergo a change in its biological characters by cell passage. Therefore, the gene ex[ressopm of exogenous gene transferred human stem cell might be affected to the epigenetic modulation such as promoter methylation and Histone deacetylation and to the cell culture conditions.
Journal of the korean academy of Pediatric Dentistry
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v.29
no.3
/
pp.345-353
/
2002
Bone morphogenetic proteins(BMPs) are secretory signal molecules which have a variety of regulatory functions during morphogenesis and cell differentiation. To evaluate roles of BMPs and their receptors on mouse sagittal suture development, we have examined their expression patterns in serial sections of sagittal sutures by in situ hybridization during embryonic stages(E15-E18). BMP-2 and BMP-3 were expressed in the osteogenic front and parietal bone on embryonic 15day, from E16 in hair follicle. BMP-4 was strongly expressed in the osteogenic front and weakly expressed in the mesenchyme and parietal bone. BMP-S was expressed in the hair follicles. BMP-6 was not expressed in this study. BMP-7 was expressed in parietal bone during embryonic stage. BMPR-IB was expressed in the osteogenic front, but BMPR-IA was not. From these datas, we suggest that the BMP-4 regulates the early commitment of mesenchymal cells to the osteogenic lineages, the BMP-2 and BMP-3 may be involved in regulating the differentiation of osteoblast precursor cells. BMP-7 was involved in maintenance of differentiated osteoblasts. BMPs were key signaling molecules that regulate early calvarial bone morphogenesis, mediated by BMPR-IB.
Park, Se-Ah;Kang, Hyun-Mi;Heo, Jin-Yeong;Yoon, Jin-Ah;Kim, Hae-Kwon
Clinical and Experimental Reproductive Medicine
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v.36
no.1
/
pp.23-34
/
2009
Objectives: Mesenchymal stem cells (MSC) comprise a promising tool for cellular therapy. It is known that long-term in vitro culture of human bone marrow and adipose tissue derived-MSCs lead to a reduction of life span and a change of stem-like characters. The aim of our study was to examine whether stem cell properties of human umbilical cord-derived stem cells (HUC) could be affected by in vitro expansion. Methods: HUC were isolated from human umbilical cord and cultured for 10 passages in vitro. Morphology and population doubling time (PDT) were investigated, and changes of stem cell properties were examined using RT-PCR and immunocytochemistry during serial subcultures. Results: Morphology and PDT of HUC began to change slightly from the 7th passage (p7). Expression level of nestin and vimentin mRNAs increased along with the culture period from p4 until p10. In contrast, expression level of SCF mRNA decreased during the same culture period. Expression level of Oct-4 and HNF-4${\alpha}$ mRNAs was not significantly changed throughout the culture period until p10. Expression level of BMP-4, FGF-5, NCAM and HLA-ABC mRNAs appeared to increase as the culture continued, however, the difference was not significant. Immunocytochemical studies showed that HUC at p3, p6 and p9 positively were stained with antibodies against SSEA-3 and SSEA-4 proteins. Interestingly, staining intensity of HUC for ICAM-1 and HLA-ABC gradually increased throughout the culture period. Intensity against thy-1 and fibronectin antibodies increased at p9 while that against TRA-1-60 and VCAM-1 antibodies began to decrease at p6 until p9. Conclusions: These results suggest that HUC change some of their stem cell characteristics during in vitro culture. Development of culture system might be needed for the maintenance of characteristics.
The aim of this essay is to illustrate Sunjung Manhwa in the 1970s which has been alienated in comics studies. This essay analyses the articles and the serial comics in Schoolgirl, the magazine in the 1970s, and examines the ideal representations of the girls at that time. Sunjung Manhwa is really different between the 1960s and 1970s. It cannot be explained on this gap just by analyzing Sunjung Manhwa in book form alone. Even though the censorship on comics was the element that has hampered the development of comics as a whole, the slumps of Sunjung Manhwa in the 1970s were very excessive compared to other comics genres. This article can gain the answers to the reason of the changes of Sunjung Manhwa by studying the magazines which was the main mass media aimed at girls with Sunjung Manhwa. While the articles in magazines show the editing direction and its characteristics, they reflect the values and ideologies at that time. The same is true for the comics in the magazines. Especially, the comics in the magazines was relatively free from the censorship. This essay examined how the articles and the comics in the girls' magazine in the 1970s represented the images of girls at the time by focusing on feature articles and comics in the magazine, Schoolgirl. This article explored Um, Hee-Ja's Blue Zone and Bang, Young-Jin's Mini March among a full-length serial comics in the magazine, Schoolgirl. Both Blue Zone and Mini March reveal the images of an ideal girl that has been emphasized by the articles in Schoolgirl. Blue Zone draws the appearances of an earnest and obedient daughter, and Mini March represents the figures of a cheerful and bright girl. Through this study, it can be recognized that the magazines in the 1970s highly appraised girls who are obedient to a given society and serve to a harmonious family as ideal ones, and it might be guessed that the ideal images of girls that was characterized ceaselessly by the magazines were the standard of the censorship on comics and its creativity and had also a huge impact on the contents and the expressions of a great deal of works. The 1970s was the times when its importance has been lost in the history of the comics studies by the censorship on the comics and the monopoly of "Hapdong(합동) publisher." The limits of expression in terms of censorship were awfully distinct, so its result was few of good works in quality, and there are still many blanks in the study on 1970s' comics. This study has a meaning which fills up a blank in the comics studies.
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