• KSBB Journal
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• 제15권5호
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• pp.489-496
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• 2000

고농도 유전자 재조합 대장균을 이용하여 pyruvate dehy-drogenase complex-E2 특이성 인간 모노클론 항체의 Fab 부분을 효율적으로 생산하기 위해 회분식, 이단 연속식, 반 유 가식, two-stage cyclic fed-batch 동 여러 가지 배양 방법이 조사되었다. 먼저 플라스미드 안정성 문제를 극복하기 위해 growth stage와 production stage를 분리하는 two-phase 회분식 배양과 이단 연속식 시스템을 시도하였다 그 결과 two-phase 회분식 배양보다는 이단 연속식 배양에서의 세포농도와 항체 생산성이 우수하였다 또한 이단 연속식 배양에서의 세포 성 장과 항처l 생산성은 용존산소를 제어한 경우가 그렇지 않은 경우보다 월등하게 높았다. 그리고 plasmid 안정성에 있어서 는 실험기간 내에 거의 100%를 유지하여 높은 안정도를 보 여주었다. 유가식 공정에 적합한 공급 배지로 변형된 M9 배 지가 최적배지로 선정되었고 이 배지 중 최적의 CjN 비율을 조사한 결파 2:3으로 결정되었다. 반 유가식 시스템에서 constant feeding 전략을 사용할 경우 최적 공급속도는 $0.6g/\ell/hr$이었다. 또한 pulse에 의해 공급배지를 공급할 경우에는 총 공급 량이 같을 경우 소량으로 자주 공급해 주는 것이 공급배지를 한꺼번에 많은 양을 공급해주는 것 보다 바람직하였다. 여러 가지 feeding 전략을 조사해 본 결과 linear feeding 방법이 가장 효과적이었다. 하지만 linear feeding 방법마저도 고농도 세포배양에 한계가 있었기 때문에 pH-stat 방법을 이용한 two-stage cyclic fed-batch 시스템을 시도하여 $54 g/\ell$의 세포 를 얻을 수 있었다. 따라서 이 방법이 일단 생산성 향상을 위한 세포의 고농도 배양에는 조사한 여러 배양 시스템 중에 가장 효율적인 시스템임올 알 수 있었다 하지만 이 시스템 에서 포도당을 낮은 level로 유지할 수 있었으나, 초산의 과도한 축적으로 항체 생산성의 향상은 예상에 비해 크지 않았다.

• Journal of Microbiology and Biotechnology
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• 제17권11호
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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.

• 한국환경복원기술학회지
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• 제7권6호
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• pp.75-83
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• 2004

본 연구는 경량형 옥상조경 소재로 도입가능성이 높은 자생식물을 중심으로 인공배합토에서 토심과 관수주기에 따른 식물의 생육반응을 검증함으로써 경량이면서 저관리형의 옥상조경을 실현하고 자생식물의 활용성을 높이고자 실시하였으며 그 결과는 다음과 같다. 1. 토심과 관수주기에 상관없이 생육이 양호하였던 수종은 애기기린초(Sedum middendorffianum Maxim)와 두메부추(Allium senescens L.)로 고사율 0%로 나타나 토심 5cm의 무관수의 조건에서도 생육이 가능한 저관리 경량형 옥상녹화에 적용 가능한 소재라 판단된다. 또한 두메부추는 토심 5cm에서 건물률이 높아 충실한 생장을 한 것으로 나타났으나 엽록소 측정을 통한 관상가치에서는 토심 10cm에서 양호한 것으로 나타나 관상가치를 고려한다면 토심 10cm 이상의 옥상녹화에 도입하는 것이 바람직한 것으로 판단된다. 2. 한라구절초(Chrysanthemum zawadskii Herb. Subsp. Coreanum (Nakai) Y. Lee Stat.)와 술패랭이(Diauthus superbus L. var. longicalycinus (Maxim) Williams)는 토심 5cm에서 고사율이 높았으며 생육이 저조하였다. 관수는 토심이 5cm인 경우 반드시 필요한 것으로 나타났으며 토심 10cm에서는 무관수에서도 양호한 생장을 하였다. 따라서 무관수의 저관리 경량형 옥상녹화를 조성하기 위해서는 최소 10cm 이상의 토심을 조성하여 식재하는 것이 바람직한 것으로 판단된다. 3. 목본류인 백리향(Thymus quinquecostatus Celak)은 토심 5cm에서 무관수이거나 3주간의 관수주기에서 90% 이상 고사하였으며 최소 1주간격으로 관수하였을 경우 생육상태가 양호하였다. 특히 토심 10cm에서는 무관수일 경우에도 생육이 우수한 것으로 나타났다. 따라서 토심 10cm에서 저관리 옥상녹화의 조성에 도입 가능한 수종으로 판단된다. 결론적으로, 본 연구에서 실험한 수종은 저관리 경량형 옥상녹화에 도입할 경우 최소 토심 10cm가 적정하며 5cm 이하의 토심에서는 최소 2주일에 1회 이상의 관수를 하는 것이 바람직한 것으로 나타났다. 특히 토심 5cm에서도 애기기린초(Sedium middendorffianum)나 두메부추(Allium senescens L.)와 같은 수종을 도입하였을 경우에는 초경량 저관리의 옥상녹화 조성이 가능한 것으로 나타났다. 본 연구는 자생초화류을 중심으로 토심과 관수와의 관계에 대한 기초적인 자료를 제시한데 그 의의가 있다고 하겠다. 이에 앞으로 다양한 토양과 수종에 대한 개발과 연구가 진행되어야 하며 특히 인공배지에서의 적정시비량과 저토심에서의 목본류에 대한 실험과 연구가 이루어져야 할 것이다.

• Journal of Oral Medicine and Pain
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• 제24권1호
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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.

• 한국임상약학회지
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• 제21권2호
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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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• 제14권5호
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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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• 제2권3호
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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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• 제64권5호
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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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• 제26권1호
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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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• 제27권6호
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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.