• Journal of Embryo Transfer
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• v.28 no.3
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• pp.281-287
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• 2013

A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.423-428
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• 2000

In 1997 when cloned sheep Dolly and soon after Polly were born, it had become head-line news because in the former the nucleus that gave rise to the lamb came from cells of six-year-old adult sheep and in the latter case a foreign gene was inserted into the donor nucleus to make the cloned sheep produce human protein, factor IX, in e milk. In the last few years, once the realm of science fiction, cloned mammals especially in livestock have become almost commonplace. What the press accounts often fail to convey, however, is that behind every success lie hundreds of failures. Many of the nuclear-transferred egg cells fail to undergo normal cell divisions. Even when an embryo does successfully implant in the womb, pregnancy often ends in miscarriage. A significant fraction of the animals that are born die shortly after birth and some of those that survived have serious developmental abnormalities. Efficiency remains at less than one % out of some hundred attempts to clone an animal. These facts show that something is fundamentally wrong and enormous hurdles must be overcome before cloning becomes practical. Cloning researchers now tent to put aside their effort to create live animals in order to probe the fundamental questions on cell biology including stem cells, the questions of whether the hereditary material in the nucleus of each cell remains intact throughout development, and how transferred nucleus is reprogrammed exactly like the zygotic nucleus. Stem cells are defined as those cells which can divide to produce a daughter cell like themselves (self-renewal) as well as a daughter cell that will give rise to specific differentiated cells (cell-differentiation). Multicellular organisms are formed from a single totipotent stem cell commonly called fertilized egg or zygote. As this cell and its progeny undergo cell divisions the potency of the stem cells in each tissue and organ become gradually restricted in the order of totipotent, pluripotent, and multipotent. The differentiation potential of multipotent stem cells in each tissue has been thought to be limited to cell lineages present in the organ from which they were derived. Recent studies, however, revealed that multipotent stem cells derived from adult tissues have much wider differentiation potential than was previously thought. These cells can differentiate into developmentally unrelated cell types, such as nerve stem cell into blood cells or muscle stem cell into brain cells. Neural stem cells isolated from the adult forebrain were recently shown to be capable of repopulating the hematopoietic system and produce blood cells in irradiated condition. In plants although the term$\boxDr$ stem cell$\boxUl$is not used, some cells in the second layer of tunica at the apical meristem of shoot, some nucellar cells surrounding the embryo sac, and initial cells of adventive buds are considered to be equivalent to the totipotent stem cells of mammals. The telomere ends of linear eukaryotic chromosomes cannot be replicated because the RNA primer at the end of a completed lagging strand cannot be replaced with DNA, causing 5' end gap. A chromosome would be shortened by the length of RNA primer with every cycle of DNA replication and cell division. Essential genes located near the ends of chromosomes would inevitably be deleted by end-shortening, thereby killing the descendants of the original cells. Telomeric DNA has an unusual sequence consisting of up to 1,000 or more tandem repeat of a simple sequence. For example, chromosome of mammal including human has the repeating telomeric sequence of TTAGGG and that of higher plant is TTTAGGG. This non-genic tandem repeat prevents the death of cell despite the continued shortening of chromosome length. In contrast with the somatic cells germ line cells have the mechanism to fill-up the 5' end gap of telomere, thus maintaining the original length of chromosome. Cem line cells exhibit active enzyme telomerase which functions to maintain the stable length of telomere. Some of the cloned animals are reported prematurely getting old. It has to be ascertained whether the multipotent stem cells in the tissues of adult mammals have the original telomeres or shortened telomeres.

• The Journal of Korean Society for School & Community Health Education
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• v.5
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• pp.71-84
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• 2004

The sexual discourse has been frequent, but the subjects of it have been limited to the young, and the sex of women and that of the old people, until now, have been overlooked. According to the data in 2,000 of the Korea Statistic Service, the older-than-sixty-five-year people consist of 37.4% men and 62.6% women. Of them, the older-than-seventy-year people consist of 33.8% men and 55.2% women. The data show the fact that as the age of the old people is higher, the number of old women is enlarging. Paying attention to the above-mentioned tendency, this study conducted a survey about the aspects of the sexual consciousness of old women. The purpose of study was focused, through the survey, on discovering the ways of old women's recognization about their sexual desire, and on presenting basic data to developing the programs which could solve the problems derived from the discovery. That is to say, the purpose of it was to prevent the sexual desire's distortion and suppression of old women, if they had any, and to present basic data to developing the programs which could convert their recognization of sex. The objects of the survey were the sample group of 301 people inhabiting in Hwaseong City, Seongnam City, Yongin City, and Suwon City, chosen out of 375,298 old women of 614,316 older-than-sixty-five-year people in Kyeonggi Province (Kyeonggi Province homepage, 2002). The data were collected by the questionnairers' interviews in the fields of old men's resting house, churches, temples, old men's niversities and so like, from September 19th to September 31th in 2003. The questionnairers as volunteers were taught the prior education, being prepared for the survey about old women's sexual consciousness. The contents of education were composed of the understanding of old women, the ways of questionnairing, and the items of questionnaire. The outcomes of the survey are following; First, concerning general items, the old women of the sample group answered that they felt greatly the difficulties of health(36.5%), economy(12.0%), and loneliness(11.3%) in turn. The 48.2% of them answered that they were healthy, but the 62.3% answered that they were suffering various illnesses. The 24.4% answered they were economically rich, and the 23.9% answered they were working. The 53.5% were living alone due to divorce or death. The 71.1% had been living for more than ten years. The 83.1% believed in their own religion. Second, the 43.8% of the sample group admitted the necessity of a heterosexual friend and sexuality The 39.0% felt sexual desire, and the 33.9% were experiencing sexuality. As the obstructing factors in performing sexuality, the objects of sample group pointed out shamefulness or morality(28.7%), anxiety that such deeds might afflict their health or the problem of health itself(20.2%), weakening of sexual potency(16.9%). This outcome shows that they have ambiguous notions about their sexual alienation and manner. Third, the 33.7% of the sample group admitted sexual education and sexual counsel, and of those that admitted them, the 85.2% said that they would take part in sexual education if it were held. It shows that the demand of sexual education and sexual counsel need be fulfilled. Therefore, through this study, the following two important suggestions can be drawn: First, systematic sexual education programs should be developed, considering old people's developing steps containing their rights of health and sex. Also those should be educated through kindergartens, primary, middle and higher education institutions. Second, sexual education and sexual counsel on prospective old people and present old people should be performed.

• Journal of Food Hygiene and Safety
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• v.26 no.4
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• pp.424-434
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• 2011

The increase in the consumption of herb medicines have made their use a public health problem due to the potential fungal contamination and the risk of the presence of my cot ox ins. 360 samples of herb medicines were evaluated for the aflatoxin contamination. The natural occurrence of aflatoxins in these samples were determined using immunoaffinity column clean up and high performance liquid chromatography (HPLC) with post-column derivatization. For samples analyzed, mean levels (incidence) of AFB1, AFB2, AFG1 and AFG2 in positive samples were $1.4\;{\mu}g/kg$(46.4%), $0.4\;{\mu}g/kg$(25.4%), $1.1\;{\mu}g/kg$(37.8%) and $0.9\;{\mu}g/kg$(24.3%), respectively. Recoveries of the full analytical procedure were 71.7~99.7% for AFB1, 88.1~99.2% for AFB2, 82.8~95.5% for AFG1 and 77.9~90.0% for AFG2. The excess cancer risk estimated using the cancer potency of aflatoxin B1 (7 $(mg/kg/day)^{-1}$ for $HBsAg^-$ and 230 $(mg/kg/day)^{-1}$ for $HBsAg^+$) were $1.30{\times}10^{-5}{\sim}1.22{\times}10^{-7}$ for hepatits B surface antigen negative ($HBsAg^-$) and $3.31{\times}10^{-4}{\sim}3.12{\times}10^{-6}$ for hepatits B surface antigen positive ($HBsAg^+$) respectively. In conclusion, although the contamination levels of samples used in the study were low, further actions are also required to undertake a program of herbal surveys in order to access mycotoxin contamination overall so that the safety of public will be protected.

• Clinical and Experimental Pediatrics
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• v.46 no.1
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• pp.56-66
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• 2003

Purpose : Human umbilical vein endothelial cells(HUVECs) play an important role in regulating blood flow by releasing vasoactive substances. It has been reported that endothelial impairment and dysfunction might be a primary cause of placental vascular disease, which is manifested clinically as preeclampsia in mother and intrauterine growth restriction in fetus. Furthermore, the frequency of apoptotic changes is increased in umbilical and placental tissues from growth-restricted pregnancies. However, the various mechanisms of umbilical endothelial cell apoptosis have not been broadly proposed. We investigate the effects of amiloride derivatives on apoptotic death of HUVECs and identify their ionic mechanism. Methods : HUVECs were purchased from Clonetics, and cultured on endothelial cell growth medium. MTT assay and flow cytometry were used for assessing cytotoxic effect and confirming the apoptosis. Changes in intracellular ion concentrations were measured with specific fluorescent dyes and fluorescence imaging analysis system. Results : Amiloride derivatives elicited cytotoxic effects on HUVECs with dose-dependent manners and the rank order of potency is HMA($IC_{50}\;11.2{\mu}M$), MIA>EIPA>>amiloride. HMA-induced cytotoxicity is dependent on extra- and intracellular pH, that is, increase extra- and intracellular pH augmented the cytotoxic effects of HMA. HMA dose-dependently reduced intracellular major ions, such as $K^+$ and $Cl^-$. Interestingly, the depletion of intracellular ions induced by HMA was also significantly enhanced at alkaline extracellular pH. Conclusion : Amiloride derivatives induce apoptosis of HUVECs with dose and pH dependent manners. They reduce intracellular $K^+$ and $Cl^-$ concentration, which is also extracellular pH dependent.

• Journal of the Korean Institute of Traditional Landscape Architecture
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• v.32 no.3
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• pp.58-68
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• 2014

Christian Cay Lorenz Hirschfeld is often regarded as 'a father of landscape garden art.' He was an aesthetics professor and garden theoretician in the $18^{th}$ century. He put forth the most comprehensive garden theory book in five volumes between 1779 and 1785. His book, Theorie der Gartenkunst, was translated and widely circulated in his contemporary. The book, which dealt with diverse aspects of garden art such as history, design, material, and type, urged to promote the prevalence of landscape garden in European continents as well as in Germany. However, there have scarcely been discourses in the Hirschfeld's garden theory. This essay aims to review Hirschfeld's garden thoughts in his book critically and to reinterpret some issues in the contemporary landscape theory and practice. Hirschfeldian theory was the product of $18^{th}$ century German Enlightenment and romanticism. At that time, Nature was regarded as divine realm. There was a German affinity with natural world. The spread of reading culture and the fashion of travel literature were another background of the success of his garden literature. Several issues in Hirschfeld garden theory will discussed here. First, privileging garden art was the most significant contribution in his theory. He emphasized that garden art was the most advanced art form among all art genres. Second, garden art was grounded on the mimesis of nature. The ambiguous relationship between nature and art still existed in garden making. However, garden art can be flourished when utilizing the potency of nature in itself. Third, there was the association between the image and the idea in experiencing the garden. Some garden scenes stimulated the related emotional responses such as cheerful and merry, softly melancholic, romantic, solemn etc. Fourth, the movement was the essential aspect of garden art. Motion and emotion are come together in garden experience. To represent the landscape garden style in suitable way, the sketch or image seems to be preferable than the plans and views. Finally, garden art was composing of not only the physical space but also the spirit of place. He maintained the garden art as hortus moralis should be a social metaphor. Hirschfeldian garden theory has often been criticized as the lack of practical power and the old fashioned idea. However, his theory influenced on formulating the idea of public park in $19^{th}$ century. Moreover, there are still some visionary aspects of his theory such as the reevaluation of garden art, the emphasis of locality and the introduction of Mittelweg idea. Recently, gardening culture are prevalent in various realms of art and life. Hirschfeld's garden theory as humanistic landscape theory can provide us some insights in the contemporary practices.

• The Korean Journal of Physiology
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• v.18 no.2
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• pp.163-169
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• 1984

The effect of vanadate on the optimum pH of Na-K-ATPase was investigated. The results were as follows: 1) The optimum PH of Na-K-ATPase was shifted from PH 7.4 to 6.8 at 10 mM K by $5{\times}10^{-6}M$ vanadate. 2) The ratio of Na-K-ATPase activity at pH 6.8 and 7.4 increased with increasing vanadate concentration. 3) Inspite of the presence of $5{\times}10^{-6}M$ vanadate Na-K-ATPase activity at pH 7.4 was higher than that at pH 6.8 below 50 mM $Na^+$, and the ratio of Na-K-ATPase activity at pH 7,4 and 6.8 was higher than that of the control. 4) Na-K-ATPase activity at pH 7.4 was higher than that at pH 6.8 below 7mM $K^+$. 5) Optimum pH of Na-K-ATPase activity was shifted from pH 7.4 to 6.8 by $10^{-5}M$ vanadate at 5 mM $K^+$. 6) $K^+$-pNPPase activity increased with lowering of pH, and the degree of inhibition of $K^+$-pNPPase activity by $10^{-7}$M vanadate was decreased with lowering of pH. These results suggest that vanadate shifts the optimum pH of Na-K-ATPase activity to more acidic PH than PH 7.4. This effect may not be caused by the decrease in the inhibitory potency of vanadate itself to Na-K-ATPase by the change of medium pH, but mainly by the alteration of Na-and K-binding site, which appears in the presence of vanadate only.

• The Korean Journal of Nuclear Medicine
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• v.32 no.3
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• pp.298-304
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• 1998

Purpose: We evaluated the usefulness of Re-188 sulfur colloid for radiation synovectomy and therapy of intraperitoneal metastasis. Materials and Methods: We investigated the labeling efficiency of Re-188 sulfur colloid on various conditions. The stability of Re-188 sulfur colloid was observed at room temperature for 24 h and in human serum and synovial fluid for 72 h. The particle size distribution of Re-188 sulfur colloid was measured by filtering with various pore size filters. Animal experiment was performed in mice and rabbits. Results: The labeling efficiency of Re-188 sulfur colloid was $64.5{\pm}5.8%$ (n=5) at the conditions of sodium thiosulfate 40 mg, EDTA $Na_2.2H_2O$ 0.8 mg, $KReO_4$ 0.8 mg at pH 1. After purification, the radiochemical purity was higher than 99%. The stability of Re-188 sulfur colloid was high (>99%) at room temperature for 24 h and in human serum and synovial fluid for 72 h. The particle size distribution of Re-188 sulfur colloid was 0.3% ($<1{\mu}m$), 11.2% ($1{\sim}5{\mu}m$), 25.8% ($5{\sim}10{\mu}m$) and 52.8% ($>10{\mu}m$). In mice, 1 h postinjection of Re-188 sulfur colloid into tail vein, uptakes in lung, liver and muscle were $37.30{\pm}5.36$, $32.33{\pm}1.79$, $6.60{\pm}0.02%$ ID/organ respectively. After i.p. injection in mice, the uptakes of extraperitonial organs of Re-188 sulfur colloid at 1 and 24 h were $0.1{\pm}0.1$, $0.4{\pm}0.1%$ ID/organ, and the excretions through urine and feces (${\sim}70 h$) were low ($2.68{\pm}0.80$, $0.95{\pm}0.17%$). When Re-188 sulfur colloid was injected to synovial space of rabbit, the uptake in other organs except knee was very low. Conclusion: Re-188 sulfur colloid showed high labeling efficiency, stability and potency for clinical use.

• Journal of Life Science
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• v.26 no.12
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• pp.1422-1430
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• 2016

This study was performed to investigate the modulatory effects of two prototypes of Panax ginseng saponin fractions, 20(S)-protopanaxadiol saponins (PDS) and 20(S)-protopanaxatriol saponins (PTS), on the induction of inflammatory mediators in lipopolysaccharide (LPS)-treated RAW264.7 murine macrophage cells. For this purpose, RAW264.7 cells were treated with LPS ($10{\mu}g/ml$) before, after, or simultaneously with PDS or PTS ($150{\mu}g/ml$), and the released level of nitric oxide (NO) and expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were evaluated. When RAW264.7 cells were treated with LPS and ginseng saponin fractions simultaneously for 24 hr, PTS, compared to PDS, more strongly attenuated the NO production induced by LPS treatment. When the cells were pretreated with LPS for 2 hr followed by PDS or PTS treatment for 24 hr, both ginseng saponins strongly reduced NO release. The pretreatment of RAW264.7 cells with PDS or PTS for 2 hr followed by LPS treatment for 24 hr significantly attenuated the LPS-induced production of NO. PTS showed stronger inhibitory potency to NO generation than PDS. Our western blot experiment showed that both PDS and PTS ($150{\mu}g/ml$) also significantly down-regulated the expressions of iNOS and COX-2 induced by LPS treatment. Our results suggest that both PDS and PTS possess strong protective effects against LPS-stimulated inflammation and that their protective effects are mediated by the suppression of NO synthesis via down-regulation of pro-inflammatory enzymes, iNOS, and COX-2 in the RAW264.7 cells.

• The Korean Journal of Nuclear Medicine
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• v.30 no.4
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• pp.507-515
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• 1996

Background : Potassium channel opener (K-opener) opens ATP-sensitive K'-channel located at cell membrane and induces potassium efflux from cytosol, resulting in intracellular hyperpolarization. Newly synthesized K-opener is currently examined for pharmacologic potency by means of rubidium release test from smooth muscle strip pre-incubated with Rb-86. Since in-vivo behavior of thallium is similar to that of rubidium, we hypothesized that K-opener can alter T1-201 kinetics in vivo. Purpose : This study was prepared to investigate the effects of pinacidil (one of potent K-openers) on the T1-201 uptake and clearance in cultured myocyte, and in-vivo biodistribution in mice. Methods : Spontaneous contracting myocytes were prepared to imitate in-vivo condition from 20 hearts of 3-5 days old Sprague-Dawley rat and cultured for 3-5 days before use ($5{\times}10^5$ cells/ml). Pinacidil was dissolved in 10% DMSO solution at a final concentration of 100nM or l0uM and was co-incubated with T1-201 in HBSS buffer for 20-min to evaluate its effect on cellular T1-uptake, or challenged to cell preparation pre-incubated with T1-201 for washout study. Two, 40 or $100{\mu}g$ of pinacidil was injected intravenously into ICR mice at 10 min after $5{\mu}Ci$ T1-201 injection, and organ uptake and whole body retention rate were measured at different time points. Results : Co-incubation of pinacidil with T1-201 resulted in a decrease in T1-201 uptake into cultured myocyte by 1.6 to 2.5 times, depending on pinacidil concentration and activity of T1-201 used. Pinacidil enhanced T1-201 washout by 1.6-3.1 times from myocyte preparations pre-incubated with T1-201. Pinacidil treatment appears to be resulted in mild decreases in blood and liver activity in normal mice, in contrast, renal and cardiac uptake were mildly decreased in a dose dependent manner. Whole body retention ratios of T1-201 were lower at 24 hour after injection with $100{\mu}g$ of pinacidil than control. Conclusion : These results suggest that treatment with K-opener may affect the interpretation of T1-201 myocardial images, due to decreasing thallium accumulation and enhancing washout from myocardium.