• International Journal of Stem Cells
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• v.16 no.3
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• pp.293-303
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• 2023

Background and Objectives: The physiological oxygen tension in fetal brains (~3%, physioxia) is beneficial for the maintenance of neural stem cells (NSCs). Sensitivity to oxygen varies between NSCs from different fetal brain regions, with midbrain NSCs showing selective susceptibility. Data on Hif-1𝛼/Notch regulatory interactions as well as our observations that Hif-1𝛼 and oxygen affect midbrain NSCs survival and proliferation prompted our investigations on involvement of Notch signalling in physioxia-dependent midbrain NSCs performance. Methods and Results: Here we found that physioxia (3% O₂) compared to normoxia (21% O₂) increased proliferation, maintained stemness by suppression of spontaneous differentiation and supported cell cycle progression. Microarray and qRT-PCR analyses identified significant changes of Notch related genes in midbrain NSCs after long-term (13 days), but not after short-term physioxia (48 hours). Consistently, inhibition of Notch signalling with DAPT increased, but its stimulation with Dll4 decreased spontaneous differentiation into neurons solely under normoxic but not under physioxic conditions. Conclusions: Notch signalling does not influence the fate decision of midbrain NSCs cultured in vitro in physioxia, where other factors like Hif-1𝛼 might be involved. Our findings on how physioxia effects in midbrain NSCs are transduced by alternative signalling might, at least in part, explain their selective susceptibility to oxygen.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.22 no.2
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• pp.164-173
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• 2000

The p16 protein is a cyclin dependent kinase inhibitor that inhibits cell cycle progression from $G_1$ phase to S phase in cell cycle. Many p16 gene mutations have been noted in many cancer-cell lines and in some primary cancers, and alterations of p16 gene function by DNA methylation have been noticed in various kinds of cancer tissues and cell-lines. There have been a large body of literature has accumulated indicating that abnormal patterns of DNA methylation (both hypomethylation and hypermethylation) occur in a wide variety of human neoplasma and that these aberrations of DNA methylation may play an important epigenetic role in the development and progression of neoplasia. DNA methylation is a part of the inheritable epigenetic system that influences expression or silencing of genes necessary for normal differentiation and proliferation. Gene activity may be silenced by methylation of up steream regulatory regions. Reactivation is associated with demethylation. Although evidence or a high incidence of p16 alterations in a variety of cell lines and primary tumors has been reported, that has been contested by other investigators. The precise mechanisms by which abnormal methylation might contribute to carcinogenesis are still not fully elucidated, but conceivably could involve the modulation of oncogene and other important regulatory gene expression, in addition to creating areas of genetic instability, thus predisposing to mutational events causing neoplasia. There have been many variable results of studies of head and neck squamous cell carcinoma(HNSCC). This investigation was studied on 13 primary HNSCC for p16 gene status by protein expression in immunohistochemistry, and DNA genetic/epigenetic analyzed to determine the incidence, the mechanisms, and the potential biological significance of its Inactivation. As methylation detection method of p16 gene, the methylation specific PCR(MSP) is sensitive and specific for methylation of any block of CpG sites in a CpG islands using bisulfite-modified DNA. The genomic DNA is modified by treatment with sodium bisulfate, which converts all unmethylated cytosines to uracil(thymidine). The primers designed for MSP were chosen for regions containing frequent cytosines (to distinguish unmodified from modified DNA), and CpG pairs near the 5' end of the primers (to provide maximal discrimination in the PCR between methylated and unmethylated DNA). The two strands of DNA are no longer complementary after bisulfite treatment, primers can be designed for either modified strand. In this study, 13 paraffin embedded block tissues were used, so the fragment of DNA to be amplified was intentionally small, to allow the assessment of methylation pattern in a limited region and to facilitate the application of this technique to samlples. In this 13 primary HNSCC tissues, there was no methylation of p16 promoter gene (detected by MSP and automatic sequencing). The p16 protein-specific immunohistochemical staining was performed on 13 paraffin embedded primary HNSCC tissue samples. Twelve cases among the 13 showed altered expression of p16 proteins (negative expression). In this study, The author suggested that low expression of p16 protein may play an important role in human HNSCC, and this study suggested that many kinds of genetic mechanisms including DNA methylation may play the role in carcinogenesis.

• The Journal of Industrial Distribution & Business
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• v.3 no.1
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• pp.7-16
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• 2012

Despite the enforcement of SSM control laws such as 'the Law of Developing the Distribution Industry (LDDI)' and 'the Law of Promoting Mutual Cooperation between Large and Small/medium Enterprises (LPMC)' stipulating the business adjustment system, the number of super-supermarkets (SSMs) has ever been expanding in Korea. In France, however, Super Centers are being regulated most strongly and directly in the whole Europe viewing that there is not a single SSM in Paris, which is emphasized to be the outcome from French government's regulation exerted on the opening of large scale retail stores. In France, the authority to approve store opening is deeply centralized and the store opening regulation is a socio-economic regulation driven by economic laws whereas EU strongly regulates the distribution industry. To control the French distribution industry, such seven laws and regulations as Commission départementale d'urbanisme commercial guidelines (CDLIC) (1969), the Royer Law (1973), the Doubin Law (1990), the Sapin Law (1993), the Raffarin Law (1996), solidarite et renouvellement urbains (SRU) (2000), and Loi de modernisation de l'économie (LME) (2009) have been promulgated one by one since the amendment of the Fontanet guidelines, through which commercial adjustment laws and regulations have been complemented and reinforced while regulatory measures have been taken. Even in the course of forming such strong regulatory laws, InterMarche, the largest supermarket chain in France, has been in existence as a global enterprise specialized in retail distribution with over 4,000 stores in Europe. InterMarche's business can be divided largely into two segments of food and non-food. As a supermarket chain, InterMarche's food segment has 2,300 stores in Europe and as a hard-discounter store chain in France, Netto has 420 stores. Restaumarch is a chain of traditional family restaurants and the steak house restaurant chain of Poivre Rouge has 4 restaurants currently. In addition, there are others like Ecomarche which is a supermarket chain for small and medium cities. In the non-food segment, the DIY and gardening chain of Bricomarche has a total of 620 stores in Europe. And the car-related chain of Roady has a total of 158 stores in Europe. There is the clothing chain of Veti as well. In view of InterMarche's management strategies, since its distribution strategy is to sell goods at cheap prices, buying goods cheap only is not enough. In other words, in order to sell goods cheap, it is all important to buy goods cheap, manage them cheap, systemize them cheap, and transport them cheap. In quality assurance, InterMarche has guaranteed the purchase safety for consumers by providing its own private brand products. InterMarche has 90 private brands of its own, thus being the retailer with the largest number of distributor brands in France. In view of its IT service strategy, InterMarche is utilizing a high performance IT system so as to obtainas much of the market information as possible and also to find out the best locations for opening stores. In its global expansion strategy of international alliance, InterMarche has established the ALDIS group together with the distribution enterprises of both Spain and Germany in order to expand its food purchase, whereas in the non-food segment, it has established the ARENA group in alliance with 11 international distribution enterprises. Such strategies of InterMarche have been intended to find out the consumer needs for both price and quality of goods and to secure the purchase and supply networks which are closely localized. It is necessary to cope promptly with the constantly changing circumstances through being unified with relevant regions and by providing diversified customer services as well. In view of the InterMarche's positive policy for promoting local partnerships as well as the assistance for enhancing the local economic structure, implications are existing for those retail distributors of our country.

• Journal of Life Science
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• v.21 no.9
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• pp.1346-1353
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• 2011

Sophora tonkinensis Gapnep has been used as a traditional herbal medicine in oriental regions since ancient times. In this study, the effect and mechanism of the MeOH extract of Sophora tonkinensis Gapnep (STME) on adipocite differentiation and adipogenesis in 3T3-L1 preadipocites were investigated. Treatment with STME in the concentration range of 0-200 ${\mu}g$/ml significantly inhibited the differentiation of 3T3-L1 preadipocites in a dose-dependent manner, as determined by a decrease in intracellular lipid droplets and lipid contents measured by Oil Red O staining. In association with the inhibitory effect of lipid accumulation, the expressions of the proteins concerned with adipogenesis in 3T3-L1 preadipocites were also investigated. Treatment with STME reduced the expressions of peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$), cytidine-cytidine-adenosine-adenosine-thymine (CCAAT)/enhancer-binding proteins ${\alpha}$ and ${\beta}$ (C/EBP${\alpha}$ and C/EBP${\beta}$) and sterol regulatory element binding protein (SREBP), which are adipocyte specific markers. In flow cytometry analysis, the inhibitory effect of differentiation was caused by G1 arrest and following mitotic clonal expansion cease. Therefore, we also investigated the alteration of G1 phase arrest-related proteins. As a result, the expression of p21 protein was significantly increased, while the expressions of Cdk2, E2F-1 and phospho-Rb were reduced in a dose-dependent manner in STME treated 3T3-L1 cells. According to these results, STME might inhibit differentiation through G1 arrest in 3T3-L1 preadipocytes adipogenesis, and further studies, which are in progress, have to be completed to identify the active compounds.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.7
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• pp.928-935
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• 2008

The myostatin gene of seven important meat (Beltex (Australia), Beltex$\times$Huyang (F1), Meat and Multi-Prolific Chinese Merino Fine Wool, Meat Chinese Merino Fine Wool and Dorper (South Africa)) and non-meat (Huyang and Kazak) sheep breeds was analyzed to study the genetic basis of muscular hypertrophy (double muscling) phenotype in sheep. SNPs, four in regulatory regions and several in the introns in the myostatin gene, were identified, and the former four SNPs were used for further studies. Twelve haplotypes were predicted by PHASE program, of which four main haplotypes (1, 3, 7, 9) were present in 90% of the 364 sheep in the study. Haplotypes 1-4 were mainly present in meat breeds while haplotypes 7 and 9 dominated the non-meat breeds. The association between haplotypes and average daily gain (ADG) was analyzed among 116 sheep with production data, Haplo2 (CGAA) and Haplo8 (TGAA) were identified to have significant (p<0.05) effect on ADG by the model (JMP5.1 software) taking into account the effects of breed, family background, haplotype, birth weight and sex. ADG of these haplotype groups also correlated well (r = 0.82) with hypertrophic phenotype scores. In conclusion, the mutations -956 (T$\rightarrow$C), -41 (C$\rightarrow$A) and 6223 (G$\rightarrow$A) involved in Haplo2 and 8 may be associated with the double-muscling trait by influencing myostatin function and be suitable markers in selecting meat sheep.

• IMMUNE NETWORK
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• v.12 no.1
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• pp.33-39
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• 2012

Background: Therapeutic approaches using monoclonal antibodies (mAbs) against complement regulatory proteins (CRPs:i.e.,CD46,CD55 and CD59) have been reported for adjuvant cancer therapy. In this study, we generated a recombinant 1E8 single-chain anti-CD59 antibody (scFv-Fc) and tested anti-cancer effect.by using complement dependent cytotoxicity (CDC). Methods: We isolated mRNA from 1E8 hybridoma cells and amplified the variable regions of the heavy chain (VH) and light chain (VL) genes using reversetranscriptase polymerase chain reaction (RT-PCR). Using a linker, the amplified sequences for the heavy and light chains were each connected to the sequence for a single polypeptide chain that was designed to be expressed. The VL and VH fragments were cloned into the pOptiVEC-TOPO vector that contained the human CH2-CH3 fragment. Then, 293T cells were transfected with the 1E8 single-chain Fv-Fc (scFv-Fc) constructs. CD59 expression was evaluated in the prostate cancer cell lines using flow cytometry. The enhancement of CDC effect by mouse 1E8 and 1E8 scFv-Fc were evaluated using a cytotoxicity assay. Results: The scFv-Fc constructs were expressed by the transfected 293T cells and secreted into the culture medium. The immunoreactivity of the secreted scFv-Fc construct was similar to that of the mouse 1E8 for CCRF-CEM cells. The molecular masses of 1E8 scFv-Fc were about 120 kDa and 55 kDa under reducing and non-reducing conditions, respectively. The DNA sequence of 1E8 scFv-Fc was obtained and presented. CD59 was highly expressed by the prostate cancer cell line. The recombinant 1E8 scFv-Fc mAb revealed significantly enhanced CDC effect similar with mouse 1E8 for prostate cancer cells. Conclusion: A 1E8 scFv-Fc construct for adjuvant cancer therapy was developed.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1520-1520
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• 2001

Food adulteration is a serious consumer fraud and a potentially dangerous practice. Regulatory authorities and food processors require a rapid, non-destructive test to accurately confirm authenticity in a range of food products and raw materials. Olive oil is prime target for adulteration either on the basis of the processing treatments used for its extraction (extra virgin vs virgin vs ordinary oil) or its geographical origin (e.g. Greek vs Italian vs Spanish). As part of an investigation into this problem, some preliminary work focused on the ability of near infrared spectroscopy to discriminate between virgin olive oils from separate regions of the Mediterranean i. e. Crete and the Peloponese. A total of 46 oils were collected: 18 originated in Crete and 28 in the Peloponese. Oils were stored in a temperature-controlled room at 2$0^{\circ}C$ prior to spectral collection at room temperature (15-18$^{\circ}C$). Samples (approximately 0.5$m\ell$) were placed in the centre of the quartz window in a camlock reflectance cell; the gold-plated baking plate was then gently placed into the cell against the glass so as to minimize the formation of air bubbles. The rear of the camlock cell was then screwed into place producing a sample thickness of 0.5mm. Spectra were recorded between 400 and 2498nm at 2nm intervals on a NIR Systems 6500 scanning monochromator. Spectral collection took place over 2-3 days. Data were analysed using both WINISI and The Unscrambler software to investigate the possibility of discriminating between the oils from Crete and the Peloponese. A number of data pre-treatments were used and discriminant models were developed using discriminant PLS (WINISI & Unscrambler) and SIMCA (Unscrambler). Despite the small number of samples involved, a satisfactory discrimination between these two oil types was achieved. Graphical examination of principal component scores for each oil type also holds out the possibility of separating oils from either Crete and the Peloponese on the basis of districts within each region. These preliminary data suggest the potential of near infrared spectroscopy to act as a screening technique for the confirmation of geographic origin of extra virgin olive oils. The sample presentation strategy adopted uses only small volumes of material and produces high quality spectra.

• Development and Reproduction
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• v.13 no.2
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• pp.79-87
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• 2009

Due to its well-known function in parturition and milk ejection, oxytocin (OT) has been considered as a 'female neurohypophyseal hormone'. Recent studies, however, clearly indicate that OT has some local roles in male reproduction via both central and peripheral routes. In experimental rodents, OT is released within distinct brain regions in response to social stimuli, and the brain OT receptor (OTR) mediated actions were strongly involved in the regulation of a variety of male behaviors such as mating-associated behaviors. In particular, OT and/or OTR knockout mice provide important clues about the molecular regulatory mechanism of the socio-sexual behaviors. Several lines of evidence also show that OT is synthesized within rodents testis, epididymis and prostate, and the presence of OTRs in these organs. In rodent testes, OT might have a role in the modulation of steroidogenesis via stimulation of the conversion of testosterone (T) to dihydrotestosterone (DHT) by 5alpha-reductase. Similar effects of OT on this androgen conversion were observed in epididymis and prostate suggesting the OT's modulatory role, such as contractility induction, in these androgen-dependent organs. In this context, further investigations on the OT's role in male CNS and reproductive organs are likely to provide better understanding on the complex socio-sexual behaviors and a platform for development of therapeutics to treat some psychological and/or andrological problems.

• Journal of Applied Biological Chemistry
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• v.55 no.4
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• pp.227-234
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• 2012

We observed that oil began to accumulate at 25 seed days after flowering (DAF) and reached the maximum potential at 35 seed DAF of oilseed rape, and the greatest weight of 100 seeds was obtained at 35 seed DAF. To survey a broad analysis of gene expression in developing embryos of Brassica napus, the Bn 300k microarray have been constructed. The Bn 300k Microarrary was designed from 80,696 unigenes clustered from 543,448 ESTs and 780 cDNA at NCBI. These arrays have been hybridized in a series of experiments with probes derived from seeds and leaf of B. napus. Approximately 8.5% of the 7,000 genes were expressed as ratios 2-fold higher in seed (25 DAF) than leaves and 0.4% at ratios 10. Also we observed that storage and cell differentiation-related genes were highly expressed at 10 DAF, whereas energy-related genes including fatty acid metabolism were increased up depending on seed maturation using Microarray, which was confirmed by reverse transcriptase polymerase chain reaction. These results suggest that B. napus arrays provide a very useful data set of seed-specific expression that can be further analyzed by examination of the promoter regions of these genes and help our understanding of the complex regulatory network in developing seeds.

• Journal of environmental and Sanitary engineering
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• v.7 no.2
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• pp.1-10
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• 1992

Since 1960's along with industrialization and urbanization, economic growth has been . achieved, however, at the same time, environmental condition has been seriously deteriorated. . Currently, volume of wastewater has been increasing at annual rate of 7% in sewage and 20% in industrial wastewater. However, the nation's sewage treatment serves only 33% of the municipal wastewater as of 1991. Major portion of air pollutants comes from combustion of oil and coal which comprise 81% of total energy use and emission gases from motor vehicles increasing at an accelerated rate. It is known that Korea generates the highest amount of waste per capta. Nevertheless, it is not sufficient to reduce the volume of waste by means of resources recovery and recycling. Recognizing the importance of global environmental problems such as ozone layer depletion, global warming and acid rain, international society has been making various efforts since the 1972 Stockholm conference. In particular, it is expected that the Rio conference which has adopted the Rio declaration and Agenda 21 will form a crucial turning point of the emerging new world order after the Cold War confrontation. To cope with such issues as domestic pollution and global environmental problems, the fundamental national policy aims at harmonizing "environmental protection and sustainable development". The Ministry of Environment has recently set up a mid-term comprehensive plan which includes annual targets for environmental protection. According to the government plan, gradual improvement of various environmental conditions and specific measures to achieve them is planned in time frame. Additional sewage treatment plants will be constructed in urban areas with the target to treat 65% of the nation's municipal sewage by 1996. Supply of clean fuels such as LNG will also be expanded starting from large cities as a cleaner substitute energy for coal and oil. In parallel with expansion of LNG, emphasis will be placed on installation of stack monitoring system. Due to the relatively limited land, government's basic policy for solid waste treatment is to develop large scale landfill facilities rather than small sized ones. Thirty three regional areas have been designated for the purpose of waste management. For each of these regions, big scale landfill site is going to be developed. To increase the rate of waste recycling the government is planning to reinforce separate collection system and to provide industries with economic incentives. As a part of meeting the changing situation on global environmental problems after UNCED, and accommodation regulatory measures stipulated in the global environmental conventions and protocols, national policy will try to alter industrial and economic structure so as to mitigate the increasing trends of energy consumption, by encouraging energy conservation and efficiency. In this regard, more attention will be given to the policy on the development of the cleaner technology. Ultimately, these policies and programs will contribute greatly to improving the current state of national public health.