• KSBB Journal
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• v.15 no.5
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• pp.489-496
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• 2000

Several culture systems including batch, two-stage CSTR, semi-fed batch, and two-stage cyclic fed-batch were investigated for the efficient production of the Fab fraction of PDC-E2 specific human monoclonal antibody using high cell density recombinant E. coli. A two-phase batch system and a two-stage continuous system were examined to overcome plasmid instability problems, by separating the growth and the production stages. The cell density and productivity of the two-stage continuous culture was better than that of the two-phase batch fermentation. In the two-stage continuous culture system with DO-stat, the cell growth and the productivity were superior to those of the system without the DO control. Also, almost total plasmid stability was maintained in the two-stage continuous culture system. Modified M9 medium was selected as an optimum feeding medium for the fed-batch process, and the optimum C/N ratio determined to be 2:3. The optimum feeding rate was $0.6g/\ell/hr$ for a constant feeding strategy in semi-fed batch system. When the feeding medium was fed by pulsing, it was observed that more frequent pulsing resulted in improved cell growth. The linear feeding method was the most efficient of the various feeding methods tested. Finally, high cell density culture using a two-stage cyclic fed batch system with pH-stat was tried because the linear feeding method showed limitations in terms of obtaining high cell densities, and a cell density of $54 g/\ell$ was achieved. It was concluded that the two-stage cyclic fed batch system was the most efficient system for high cell density culture of the systems tested. However, productivity improvements were lower than expected due to the extremely high accumulations of acetate, although the low levels of residual glucose were maintained.

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1898-1903
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• 2007

We attempted to modulate the overall protein expression rate through the addition of a repressor against the araBAD promoter system of Escherichia coli, in which glucose was used as a repressor. Therefore, 0.5% L-arabinose was initially contained as an inducer in culture medium, and either 2% glucose or 2% glycerol was used as a carbon source, and it was found that the expression of recombinant interferon-${\alpha}$ could be observed at the beginning of the batch culture when glycerol was used as a carbon source. However, when glucose was used, the initiation of recombinant interferon-${\alpha}$ expression was delayed compared with that when glycerol was used. Furthermore, when the addition of 0.5% glucose was carried out once or twice after 0.5% L-arabinose induction during DO-stat fed-batch culture, the distributions of soluble and insoluble recombinant interferon-${\alpha}$ were modulated. When glucose was not added after the induction of L-arabinose, all of the expressed recombinant interferon-${\alpha}$ formed an inclusion body during the later half of culturing. However, when glucose was added after induction, the expressed recombinant interferon-${\alpha}$ did not all form an inclusion body, and about half of the total recombinant interferon-${\alpha}$ was expressed in a soluble form. It was deduced that the addition of glucose after the induction of L-arabinose might lower the cAMP level, and thus, CAP (catabolite activator protein) might not be activated. The transcription rate of recombinant interferon-${\alpha}$ in the araBAD promoter system might be delayed by the partial repression. This inhibition of the transcription rate probably resulted in more soluble interferon-${\alpha}$ expression caused by the reduction of the protein synthesis rate.

• Journal of the Korean Society of Environmental Restoration Technology
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• v.7 no.6
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• pp.75-83
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• 2004

Focusing on native plants that have high possibility of being introduced as rooftop material, this study was conducted to investigate extensive and easy-to-manage rooftop garden and to raise the utilization of native plants by verifying their growing response to artificial substrate soil depth and irrigation period. The study was conducted from March to September in 2002. Plants tested included Chrysanthemum zawadskii, Sedium middendorffianum, Thymus quinquecostatus, Allium senescens, and Dianthus superbus. Regarding soil depth, it was 5 cm and 10 cm. Irrigation period was non-irrigation, 1-week, 2-weeks, and 3- weeks, Its result is as follows; 1. In case of Sedum middendorffianum Maxim, mortality rate was 0% regardless of soil depth and irrigation period making it very suitable material for rooftop garden. 2. In case of Allium senescens L., mortality rate was 0% regardless of soil depth and irrigation period making it very suitable material for rooftop garden. Therefore, Provided that fertilizing is managed well, it is a plant that can be highly utilized.3. In case of Chrysanthemum zawadskii Herb. Subsp. (Nakai) Y. Lee Stat., the growth of top was lower in 10cm than in 5cm and it grew well in 10cm. When utilizing for rooftop garden, it would be desirable to keep minimum viable soil depth at over 10cm. If there is enough rainfall, soil and soil depth seem to have greater effect on growth than irrigation period does. 4. In case of Diauthus superbus L. var. longicalycinus (Maxim) Williams, rooting rate and growth were better in 10cm than in 5cm. Therefore, it is desirable to keep minimum soil depth at over 10cm. 5. In case of Thymus quinquecostatus Celak, the growth of top and flowering were better in 10cm than in 5cm. Therefore, it seems desirable to have minimum viable soil depth to be over 10cm. In conclusion, the most suitable species for rooftop garden are Sedium middendorffianum and Allium senescens in this experiment. However, Chrysanthemum zwadskii, Thymus quinquecostatus, and Dianthus chinensis also can be utilized greatly when irrigation is managed regularly in artificial mixed soil over 10cm.

• Journal of Oral Medicine and Pain
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• v.24 no.1
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• pp.95-106
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• 1999

This study was performed to investigate the age distribution with tooth calcification and degree of eruption of permanent teeth. For the study, healthy 184 patients from 5 to 19 years old without any previous serious dental treatment were randomly selected, and intraoral standard films and dental casts were taken for evaluation of stage of calcification and degree of eruption, respectively. Tooth calcification of 13 stages, designed by the author based on the Nolla's classification and eruption level of 4 or 5 degree was used. Data were processed by SAS/Stat program and the obtained results were as follows; 1. The age of root completed with open apex in lower posterior teeth were 13.8 years for first premolar, 14.0 years for second premolar, 10.5 years for first molar, and 14.2 years for second molar. There were no significant difference between right and left side. 2. As for the sequence of eruption, first molar was the first teeth erupted in upper arch, while central incisor was the first teeth in lower arch. In general, eruption of lower teeth were slightly earlier than the corresponding teeth of upper arch. 3. There were no difference of age of the same stage of development between Nolla's and the author's classification. From the results, the author's classification can be used for estimation of age with more finely in age of 8 to 15 years old. 4. Multiple regression equations for age with Nolla's(Ns) and the author's(Ks) classification of tooth calcification, and degree of eruption(DE) were as follow; Age(by #34) = 7.55 + 0.76Ks34 + 0.80DE34 - 0.72Ns34 Age(by #35) = 7.10 + 0.81Ks35 + 0.6IDE35 Age(by #37) = 6.61 + 0.82Ks37 + 0.5IDE37. Age(by #44) = 7.02 + 0.62Ks44 + 0.82DE44 Age(by #45) = 8.04 + 0.93Ks45 + 0.64DE45 - 0.89Ns45 Age(by #47) = 6.40 + 0.86Ks47 + 0.56DE47.

• Korean Journal of Clinical Pharmacy
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• v.21 no.2
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• pp.131-137
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• 2011

The purpose of this study was to evaluate the clinical efficacy of a standardized special ethanol extract from Gynostemma pentaphyllum as a management for anxiety and stress of normal population. This is a two-arm, parallelgroup, randomized, double blind clinical trial comparing Gynostemma pentaphyllum extract 200 mg bid (GP-EX, n=48) or placebo bid (n=54). The main outcome measures were the decrease in anxiety sensitivity index (ASI), the State version (S-STAI) of the Stait-Trait Anxiety Inventory (STAI) and the Trait version (T-STAT) of the STAI from baseline over a 6 weeks treatment period. In more anxious group (S-STAI50 or ASI19), the anxiety in group with GP-EX was decreased significantly than one in normal population with placebo [S-STAI50: T-STAI = from $57.7{\pm}6.5$ ($mean{\pm}S.D.$) to $46.8{\pm}11.2$ in normal population with GP-EX, p=0.002 vs. from $54.1{\pm}9.9$ to $49.0{\pm}9.6$ in normal population with placebo, p>0.05; ASI19: T-STAI = from $47.2{\pm}12.0$ to $42.4{\pm}11.1$ in normal population with GP-EX, p=0.022 vs. from $48.7{\pm}11.5$ to $46.0{\pm}10.4$ in normal population with placebo, p>0.05]. The most frequently reported adverse reactions considered possibly related to treatment were mild gastrointestinal events. GP-EX is more effective than placebo and is well tolerated as a therapy for anxiety and stress of normal population.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.5
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• pp.2689-2698
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• 2013

Epithelial-to-mesenchymal transition (EMT) is a collection of events that allows the conversion of adherent epithelial cells, tightly bound to each other within an organized tissue, into independent fibroblastic cells possessing migratory properties and the ability to invade the extracellular matrix. EMT contributes to the complex architecture of the embryo by permitting the progression of embryogenesis from a simple single-cell layer epithelium to a complex three-dimensional organism composed of both epithelial and mesenchymal cells. However, in most tissues EMT is a developmentally restricted process and fully differentiated epithelia typically maintain their epithelial phenotype. Recently, elements of EMT, specially the loss of epithelial markers and the gain of mesenchymal markers, have been observed in pathological states, including epithelial cancers. Increasing evidence has confirmed its presence in human colon during colorectal carcinogenesis. In general, chronic inflammation is considered to be one of the causes of many human cancers including colorectal cancer(CRC). Accordingly, epidemiologic and clinical studies indicate that patients affected by ulcerative colitis and Crohn's disease, the two major forms of inflammatory bowel disease, have an increased risk of developing CRC. A large body of evidence supports roles for the SMAD/STAT3 signaling pathway, the NF-kB pathway, the Ras-mitogenactivated protein kinase/Snail/Slug and microRNAs in the development of colorectal cancers via epithelial-tomesenchymal transition. Thus, EMT appears to be closely involved in the pathogenesis of colorectal cancer, and analysis refered to it can yield novel targets for therapy.

• Molecular & Cellular Toxicology
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• v.2 no.3
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• pp.193-201
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• 2006

Cephalexin, one of most widely prescribed cephalosporin, has been reported to cause acute renal failure as a side effect in human and experimental animals. Although numerous animal studies have been reported for the cephalosporin nephrotoxicity, the molecular and cellular nephrotoxic mechanisms of cephalexin are still unknown. This investigation evaluated the time-dependent gene expression profile of kidney in mouse during cephalexin induced nephrotoxicity. C57BL/6 female mice were administered either saline or 1,000 mg/kg cephalexin intraperitoneally. Mice were sacrificed at 3, 6, and 24 hr after administration. Blood biochemical and histopathological results indicated cephalexin induced nephrotoxicity. Microarray experiment carried out using Affymetrix $GeneChip^{(R)}$. There were 198 informative genes that were significantly expressed >5-fold versus control at 3, 6, and 24 hr (p<0.01), of which 156 and 42 were up-and down-regulated, respectively. Major classes of up-regulated genes at 3, 6 hr included those involved in MAPK/Jak-STAT signaling pathway and immune response such as cytokine-cytokine receptor interaction and complement and coagulation cascades. At 24 hr, up-regulated genes were mainly involved in regeneration/repair and immune response; down-regulated genes were generally associated with transporters and intermediary metabolism. Among the up-regulated genes at 24 hr, several potential biomarkers on nephrotoxicity such as Kim-1, Fga, Timp1, and Slc34a2 were clustered in a same category. In addition, Tnfrsf12a and Lcn2 which were consistently up-regulated (>5 fold) were also included as potential biomarkers. These results may provide clues for elucidating the mechanism of cephalexin induced nephrotoxicity and evaluating potential biomarkers to assess nephrotoxicity.

• Journal of Animal Science and Technology
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• v.64 no.5
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• pp.922-936
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• 2022

5-Hydroxytryptamine (5-HT), a monoamine, as a local regulator in the mammary gland is a chemical signal produced by the mammary epithelium cell. In cows, studies have shown that 5-HT is associated with epithelial cell apoptosis during the degenerative phase of the mammary gland. However, studies in other tissues have shown that 5-HT can effectively promote cell viability. Whether 5-HT could have an effect on mammary cell viability in dairy cows is still unknown. The purpose of this study was to determine: (1) effect of 5-HT on the viability of bovine mammary epithelial cells and its related signaling pathways, (2) interaction between prolactin (PRL) and 5-HT on the cell viability. The bovine mammary alveolar cell-T (MAC-T) were cultured with different concentrations of 5-HT for 12, 24, 48 or 72 hours, and then were assayed using cell counting kit-8, polymerase chain reaction (PCR) and immunobloting. The results suggested that 20 μM 5-HT treatment for 12 or 24 h promote cell viability, which was mainly induced by the activation of 5-HT receptor (5-HTR) 1B and 4, because the increase caused by 5-HT vanished when 5-HTR 1B and 4 was blocked by SB224289 and SB204070. And protein expression of mammalian target of rapamycin (mTOR), eukaryotic translation elongation factor 2 (eEF2), janus kinase 2 (JAK2) and signal transducer and activator of transcription 5 (STAT5) were decreased after blocking 5-HT 1B and 4 receptors. When MAC-T cells were treated with 5-HT and PRL simultaneously for 24 h, both the cell viability and the level of mTOR protein were significantly higher than that cultured with 5-HT or PRL alone. In conclusion, our study suggested that 5-HT promotes the viability of MAC-T cells by 5-HTR 1B and/or 4. Furthermore, there is a reciprocal relationship between PRL and 5-HT.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.1
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• pp.37-45
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• 2022

The aim of the present study was to investigate the physiological role of nicotinamide phosphoribosyltransferase (NAMPT) associated with odontogenic differentiation during tooth development in mice. Mouse dental papilla cell-23 (MDPC-23) cells cultured in differentiation media were stimulated with the specific NAMPT inhibitor, FK866, and Visfatin (NAMPT) for up to 10 days. The cells were evaluated after 0, 4, 7, and 10 days. Cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The mineralization assay was performed by staining MDPC-23 cells with Alizarin Red S solution. After cultivation, MDPC-23 cells were harvested for quantitative PCR or Western blotting. Analysis of variance was performed using StatView 5.0 software (SAS Institute Inc., Cary, NC, USA). Statistical significance was set at p < 0.05. The expression of NAMPT increased during the differentiation of murine odontoblast-like MDPC-23 cells. Furthermore, the up-regulation of NAMPT promoted odontogenic differentiation and accelerated mineralization through an increase in representative odontoblastic biomarkers, such as dentin sialophosphoprotein, dentin matrix protein-1, and alkaline phosphatase in MDPC-23 cells. However, treatment of the cells with the NAMPT inhibitor, FK866, attenuated odontogenic differentiation, as evidenced by the suppression of odontoblastic biomarkers. These data indicate that NAMPT regulated odontoblastic differentiation through the regulation of odontoblastic biomarkers. The increase in NAMPT expression in odontoblasts was closely related to the formation of the extracellular matrix and dentin via the Runx signaling pathway. Therefore, these data suggest that NAMPT is a critical regulator of odontoblast differentiation during tooth development.

• Restorative Dentistry and Endodontics
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• v.27 no.6
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• pp.612-621
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• 2002

The purpose of this study was to compare the histomorphological change of curved root canal preparation using GT rotary File, Profile .04 taper and stainless steel K-file. 45 mesial canals(over 20 degree) of extracted human mandibular first molars were mounted in resin using a modified Bramante muffle system and divided into three groups. The roots were cross-sectioned at 2.5mm 5mm and 8mm levels from apical foramen. Tracings of the canals were made from preinstrumentation pictures of the cross section. The canals were prepared using a step-back technique with stainless steel K file(group 1), Profile .04 taper rotary file(group 2) and GT rotary file(group 3). Tracings of the prepared canals were made from postinstrumentation picture. Canal centring ratio. amount of transportation, area of dentin removed and shape of canal were measured and statistically were evaluated with Student-Newman-Keuls test using Sigma Stat(Jandel Scientific Software, USA). The results were as followings : 1 Amount of transportation of group 2 was the lowest at apical part, but there was no statistical difference. The direction of transportation was the outside of curvature at apical part. 2. Centering ratio at the apical part of group 1 was the highest, and there was statistical differences between apical and middle part, apical and coronal part(p＜0.05). Centering ratio at the middle part of group 3 was the lowest, and there was statistical difference between apical and middle part(p＜0.05). Centering ratio of group 2 was the lowest at apical part, but there was no statistical difference. 3. Amount of dentin removed of group 1 was the highest at coronal, middle and apical part among three groups, and there was statistical difference(p＜0.05). 4. The majority of the cross-sectioned canal shape after instrumentation were irregular at coronal, middle and apical part. But there are more number of round shaped canals at group 3 than other group.