• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1015-1025
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• 2015

The development of diverse polymyxin derivatives is needed to solve the toxicity and resistance problems of polymyxins. However, no platform has generated polymyxin derivatives by genetically engineering a polymyxin synthetase, which is a nonribosomal peptide synthetase. In this study, we present a two-step approach for the construction of engineered polymyxin synthetases by substituting the adenylation (A) domains of polymyxin A synthetase, which is encoded by the pmxABCDE gene cluster of Paenibacillus polymyxa E681. First, the seventh L-threonine-specific A-domain region in pmxA was substituted with the L-leucine-specific A-domain region obtained from P. polymyxa ATCC21830 to make polymyxin E synthetase, and then the sixth D-leucine-specific A-domain region (A_6-D-Leu-domain) was substituted with the D-phenylalanine-specific A-domain region (A_6-D-Phe-domain) obtained from P. polymyxa F4 to make polymyxin B synthetase. This step was performed in Escherichia coli on a pmxA-containing fosmid, using the lambda Red recombination system and the sacB gene as a counter-selectable marker. Next, the modified pmxA gene was fused to pmxBCDE on the chromosome of Bacillus subtilis BSK4dA, and the resulting recombinant strains BSK4-PB and BSK4-PE were confirmed to produce polymyxins B and E, respectively. We also succeeded in constructing the B. subtilis BSK4-PP strain, which produces polymyxin P, by singly substituting the A_6-D-Leu-domain with the A_6-D-Phe-domain. This is the first report in which polymyxin derivatives were generated by genetically engineering polymyxin synthetases. The two recombinant B. subtilis strains will be useful for improving the commercial production of polymyxins B and E, and they will facilitate the generation of novel polymyxin derivatives.

• BMB Reports
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• 제50권11호
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• pp.537-538
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• 2017

Hypoxia affects various physiological and pathophyological processes. Hypoxia changes the expression of hypoxia-responsive genes through two main pathways. First, hypoxia activates transcription factors (TF) such as Hypoxia-inducible Factor (HIF). Second, hypoxia decreases the activity of Jumonji C domain-containing histone demethylases (JMJDs) that require $O_2$ and ${\alpha}$-Ketoglutarate (${\alpha}$-KG) as substrates. The JMJDs affect gene expression through their regulation of active or repressive histone methylations. Profiling of H3K4me3, H3K9me3, and H3K27me3 under both normoxia and hypoxia identified 75 TFs whose binding motifs were significantly enriched in the methylated regions of the genes. TFs showing similar binding strengths to their target genes might be under the 'metabolic control' which changes histone methylation and gene expression by instant changing catalytic activities of resident histone demethylases.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.326.1-326.1
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• 2002

We applied serological analysis of cDNA expression library technique to identify cancer-associated genes. We screened cDNA expression libraries of human testis and gastric cancer cell lines with sera of patients with gastric cancers. We identified a gene whose expression is testis-specific among normal tissues. We cloned and characterized this novel gene. It contains D-E-A-D box domain and encodes a putative protein of 630 amino acids with possible helicase activity. It showed wide expression in various cancer tissues and cancer cell lines. (omitted)

• 한국지능시스템학회:학술대회논문집
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• 한국퍼지및지능시스템학회 2003년도 ISIS 2003
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• pp.447-450
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• 2003

In this research, we propose an automatic knowledge acquisition and composite knowledge expression mechanism based on machine learning and relational database. Most of traditional approaches to develop a knowledge base and inference engine of expert systems were based on IF-THEN rules, AND-OR graph, Semantic networks, and Frame separately. However, there are some limitations such as automatic knowledge acquisition, complicate knowledge expression, expansibility of knowledge base, speed of inference, and hierarchies among rules. To overcome these limitations, many of researchers tried to develop an automatic knowledge acquisition, composite knowledge expression, and fast inference method. As a result, the adaptability of the expert systems was improved rapidly. Nonetheless, they didn't suggest a hybrid and generalized solution to support the entire process of development of expert systems. Our proposed mechanism has five advantages empirically. First, it could extract the specific domain knowledge from incomplete database based on machine learning algorithm. Second, this mechanism could reduce the number of rules efficiently according to the rule extraction mechanism used in machine learning. Third, our proposed mechanism could expand the knowledge base unlimitedly by using relational database. Fourth, the backward inference engine developed in this study, could manipulate the knowledge base stored in relational database rapidly. Therefore, the speed of inference is faster than traditional text -oriented inference mechanism. Fifth, our composite knowledge expression mechanism could reflect the traditional knowledge expression method such as IF-THEN rules, AND-OR graph, and Relationship matrix simultaneously. To validate the inference ability of our system, a real data set was adopted from a clinical diagnosis classifying the dermatology disease.

• 한국수정란이식학회지
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• 제30권4호
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• pp.277-282
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• 2015

Cryopreservation affects osmotic tolerance and intracellular ion concentration through changes in expression levels of water and ion channels. Control of these changes is important for cell survival after cryopreservation. Relatively little is known about changes in $K^+$ channel expression compared to water channel expression. This study was performed to investigate changes in TASK-2 channel (KCNK5: potassium channel, subfamily K, member 5), a member of two-pore domain $K^+$ channel family, in cryopreserved mouse ovaries. Cryopreservation increased TASK-2 mRNA expression in mouse ovaries. In addition, TASK-2 protein expression was upregulated in vitrified and slowly frozen ovaries. TASK-2 protein was expressed in all area of granulosa cells that surround the oocyte within the follicle, except nucleus. Viability of cells overexpressed with TASK-2 was higher than that of vector-transfected cells. Our results found that TASK-2 expression was increased by cryopreservation and overexpression of TASK-2 decreased cryopreservation-induced cell death. These results suggest that TASK-2 upregulation might reduce cryodamage.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.165-171
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• 2013

The aim of the present study was to analyze the expression of FHIT and WWOX in nasopharyngeal carcinoma (NPC) and correlations with clinical pathologic features. mRNA expression of the FHIT and WWOX was assessed by real-time fluorescent relatively quantitative PCR in 61 NPC tissues and 45 non-cancerous nasopharyngeal tissues. As a result, mRNA expression levels of both FHIT and WWOX were significantly lower in NPC patients than in control samples (P=0.049 and 0.045, respectively). Moreover, the mRNA expression of both had an inverse relation with larger invasive range (P=0.035 and 0.048, respectively), poor histologic differentiation (P=0.012 and 0.016) and advanced clinical stage (P=0.026 and 0.038). Consistency was found between expression of FHIT and WWOX in the same NPC tissues (r=0.681, P=0.00). In conclusion, synergy between FHIT and WWOX may exist in the development of NPC so that the two factors may be considered as important genetic markers. Detecting the expression of FHIT and WWOX should provide clinically significant information relevatn to tumor diagnosis, progression and treatment modalities for NPC.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.4065-4069
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• 2015

Background: Epidermal growth factor-like domain multiple 7 (EGFL7), a secreted protein specifically expressed by endothelial cells during embryogenesis, recently was identified as a critical gene in tumor metastasis. Epithelial-mesenchymal transition (EMT) was found to be closely related with tumor progression. Accordingly, it is important to investigate the migration and EMT change after knock-down of EGFL7 gene expression in human pancreatic cancer cells. Materials and Methods: EGFL7 expression was firstly testified in 4 pancreatic cancer cell lines by real-time polymerase chain reaction (Real-time PCR) and western blot, and the highest expression of EGFL7 was found in PANC-1 cell line. Then, PANC-1 cells transfected with small interference RNA (siRNA) of EGFL7 using plasmid vector were named si-PANC-1, while transfected with negative control plasmid vector were called NC-PANC-1. Transwell assay was used to analyze the migration of PANC-1 cells. Real-time PCR and western blotting were used to detect the expression change of EGFL7 gene, EMT markers like E-Cadherin, N-Cadherin, Vimentin, Fibronectin and transcription factors like snail, slug in PANC-1, NCPANC-1, and si-PANC-1 cells, respectively. Results: After successful plasmid transfection, EGFL7 gene were dramatically knock-down by RNA interference in si-PANC-1 group. Meanwhile, migration ability decreased significantly, compared with PANC-1 and NC-PANC-1 group. Meanwhile, the expression of epithelial phenotype marker E-Cadherin increased and that of mesenchymal phenotype markers N-Cadherin, Vimentin, Fibronectin dramatically decreased in si-PANC-1 group, indicating a reversion of EMT. Also, transcription factors snail and slug decreased significantly after RNA interference. Conclusions: Current study suggested that highly-expressed EGFL7 promotes migration of PANC-1 cells and acts through transcription factors snail and slug to induce EMT, and further study is needed to confirm this issue.

• 한국초등수학교육학회지
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• 제17권1호
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• pp.105-128
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• 2013

우리나라 교육과정은 7차 이후 2007, 2009 개정 교육과정을 거치면서 다양한 변화를 모색하고 있다. 본 연구는 그 가운데 초등수학 교과서의 내용 변화에 주목하고 있으며, 특히 초등수학에서 다루어지고 있는 문자와 식에 초점을 맞추고 있다. 문자와 식은 6차 교육과정에서는 '관계' 영역에서, 7차 교육과정에서는 '문자와 식' 영역에서, 그리고 2007 교육과정에서는 '규칙성과 문제해결' 영역에서 다루어져왔다. 특히 7차 교육과정에서는 초등수학에서 문자가 도입되지 않았으나, 6차와 2007년 교육과정에서는 초등수학에서 문자 x의 도입, 등식의 성질, 방정식 등이 다루어지고 있다. 본 연구는 초등수학에서 이러한 변화를 겪고 있는 문자와 식에 대하여 교육과정별 교과서에 제시된 문자와 식의 내용 및 지도 시기, 지도 방법에 대한 분석을 목적으로 한다. 이를 위해 문자 x의 도입, 등식의 지도, 방정식의 지도와 같이 3가지 주제를 구분하고, 이들 각각에서 초등수학을 중심으로 6차 교육과정과 2007년 교육과정을 비교하고, 동시에 그 사이에 놓여 있는 7차 교육과정에서는 중학교 7-가 단계를 살펴보았다. 본 연구는 이를 통해 초등수학에서 문자와 식을 이해하고 지도하는데 기초자료가 될 수 있기를 기대한다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제33권3호
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• pp.191-198
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• 2007

The p53 which is well known as tumor suppressor gene is located at 17p13. p53 is a sequence-specific DNA binding transcription factor that responds to certain cytotoxic stresses, such as DNA damage, by enhancing the transcription of genes that regulate cell-cycle progression as well as programmed cell death. The p63 gene that is located at 3q27-29, is recognized members of the p53 family, and responsible for the transcription of 6 isoforms. Three isoforms ($TAp63{\alpha}$, $TAp63{\beta}$, $TAp63{\gamma}$) contain an N-terminal transactivation (TA) domain and can induce apoptosis. The other 3 isoforms (${\Delta}Np63{\alpha}$, ${\Delta}Np63{\beta}$, ${\Delta}Np63{\gamma}$) lack the TA domain and may function in a dominant-negative fashion by inhibiting the transactivation functions of p53 and TAp63 proteins, and thus act as oncoproteins. A number of studies have investigated the role of p63 in human squamous cell carcinomas from different organs. Only a few studies have examined ${\Delta}Np63$ isoform in oral squamous cell carcinoma including normal epithelium. This study aimed to evaluate expression of ${\Delta}Np63$ isoform in human oral squamous cell carcinoma tissue and normal mucosa. The 3 cases of well differenciated oral squamous cell carcinoma specimen including adjacent normal mucosa were examined, and immunohistochemical study with monoclonal antibody(4A4) and tumor cell apoptosis analysis with Transmission Electon Microscopy were studied. And, RT-PCR analysis was done for expression of ${\Delta}Np63$ isoform. The results were as followed. 1. Normal gingiva showed the restricted p63 expression in basal cell layer. 2. Well differentiated squamous cell carcinoma showed mainly p63 expression in overall area of malignancy, especially in basal cell layer to adjacent stromal tissue. 3. Tumor cells around keratinized area with no p63 expression disclosed less micro-organelle in decreased size cytoplasm and severe chromatin margination with nuclear destruction that means apoptosis. 4. Comparison of mRNA expression of ${\Delta}Np63$ isoform by RT-PCR showed variable expression of ${\Delta}Np63$ isoform, but ${\Delta}Np63{\alpha}$ was most highly expressed in all 3 tumor specimen. From theses results, it should be suggested that ${\Delta}Np63$ isoform expression in well differentiated squamous cell carcinoma was closely related to tumor oncogenesis, expecially overexpression of ${\Delta}Np63{\alpha}$ is a most important factor in tumor genesis of oral squamous cell carcinoma.

• 한국인터넷방송통신학회논문지
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• 제22권4호
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• pp.81-86
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• 2022

본 연구에서 csv포맷으로 구조화된 파일 데이터의 컬럼의 도메인을 자동 판별하는 방법을 제안한다. 데이터와 데이터 간 융합을 통해 새로운 데이터를 생성할 수 있고, 이들 새로운 데이터가 중요한 자원이 되기 위해서는 조인 되는 컬럼의 일관성이 유지되어야 한다. 데이터 품질을 측정하기 위한 방법 중의 하나가 도메인 기반 품질 진단 방법이다. 도멘인이란 각 컬럼의 성격을 규정하는 가장 광범위한 지표이므로 이를 자동으로 판별하는 방법이 필요하다. 기존의 연구에서는 관계형 데이터베이스의 도메인 자동 판별이 주로 연구 되었지만 본 연구는 파일데이터의 특성을 이용하여 도메인을 자동화 할 수 있는 모델을 개발하였다. 파일데이터의 도메인 판별을 특화하기 위하여 정규표현식을 이용하여 데이터를 단순화 하고 이를 패턴화 하였고, 컬럼명에 해당하는 데이터 헤더의 내용을 분석하여 사용된 접미사를 분석하여 파생변수로 사용하였다. 정규표현식과 접미사의 파생변수를 추가하였을 때 기존 방법인 87%의 정확도 보다 큰 95%의 정확도로 도메인을 자동 판별하는 결과를 도출하였다. 본 연구는 공공데이터 품질진단에 자동화 방법론을 제시하여 품질 측정 기간 및 인원을 줄일 수 있을 것으로 기대된다.