• Title/Summary/Keyword: Expression Strategies

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Genetically Encoded Biosensor Engineering for Application in Directed Evolution

  • Yin Mao;Chao Huang;Xuan Zhou;Runhua Han;Yu Deng;Shenghu Zhou
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1257-1267
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    • 2023
  • Although rational genetic engineering is nowadays the favored method for microbial strain improvement, building up mutant libraries based on directed evolution for improvement is still in many cases the better option. In this regard, the demand for precise and efficient screening methods for mutants with high performance has stimulated the development of biosensor-based high-throughput screening strategies. Genetically encoded biosensors provide powerful tools to couple the desired phenotype to a detectable signal, such as fluorescence and growth rate. Herein, we review recent advances in engineering several classes of biosensors and their applications in directed evolution. Furthermore, we compare and discuss the screening advantages and limitations of two-component biosensors, transcription-factor-based biosensors, and RNA-based biosensors. Engineering these biosensors has focused mainly on modifying the expression level or structure of the biosensor components to optimize the dynamic range, specificity, and detection range. Finally, the applications of biosensors in the evolution of proteins, metabolic pathways, and genome-scale metabolic networks are described. This review provides potential guidance in the design of biosensors and their applications in improving the bioproduction of microbial cell factories through directed evolution.

Identification of Drought Tolerant Genotypes by Evaluating Morpho-physiological Traits in Pepper

  • Kyu Kyu Thin;Alebel Mekuriaw;Hyerim Do;Inhwa Yeam;Je Min Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.29-29
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    • 2022
  • The fast-changing climatic conditions make plants to be vulnerable to many abiotic stresses. Drought stress is one of the limiting factors that affect pepper production in water deficient regions. It affects plant growth and development by altering physiological, morphological, and metabolic processes. Breeding drought tolerant varieties is one of the mitigation strategies to overcome the ever increasing drought disaster. Hence, screening of new drought tolerant pepper genotypes is essential. The current study was aimed to identify new drought tolerant genotypes among the collection of pepper genetic resources. In total, 70 pepper genotypes were screened for drought tolerance after exposure to drought stress condition. The pepper genotypes were classified as highly tolerant, intermediate, or severely sensitive to drought stress based on the phenotypic analysis. Consequently, 13 genotypes significantly exhibited higher recovery rate after drought stress and were classified as highly tolerant. Comparative analysis of morphological and physiological parameters and expression of drought responsive genes between tolerant and susceptible pepper genotypes will be presented and discussed. The identified tolerant genotypes will be useful resources for breeding drought tolerant pepper cultivars.

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Anticancer Drugs at Low Concentrations Upregulate the Activity of Natural Killer Cell

  • Hyeokjin Kwon;Myeongguk Jeong;Yeeun Kim;Go-Eun Choi
    • Biomedical Science Letters
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    • v.29 no.3
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    • pp.178-183
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    • 2023
  • Natural killer (NK) cells are innate cytotoxic lymphoid cells that actively prevent neoplastic development, growth, and metastatic dissemination in a process called cancer immunosurveillance. Regulation of the cytotoxic activity of NK cells relies on integrated interactions between inhibitory receptors and numerous activating receptors that act in tandem to eliminate tumor cells efficiently. Conventional chemotherapy is designed to produce an anti-proliferative or cytotoxic effect on early tumor cell division. Therapies designed to kill cancer cells and simultaneously maintain host anti-tumor immunity are attractive strategies for controlling tumor growth. Depending on the drug and dose used, several chemotherapeutic agents cause DNA damage and cancer cell death through apoptosis, immunogenic cell death, or other forms of non-killing (i.e., mitotic catastrophe, senescence, autophagy). Among stress-induced immunostimulatory proteins, changes in the expression levels of NK cell activating and inhibitory ligands and tumor cell death receptors play an important role in the detection and elimination by innate immune effectors including NK cells. Therefore, we will address how these cytotoxic lymphocytes sense and respond to high and low concentrations of drug-induced stress to the drug cisplatin, among the various types of drugs that contribute to their anticancer activity.

Study on the Symbolic Meaning of Fashion and Art Collaboration

  • JEON SUR
    • International Journal of Advanced Culture Technology
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    • v.12 no.3
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    • pp.290-302
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    • 2024
  • The collaboration between art and fashion goes beyond simple marketing strategies and creates new cultural and aesthetic values through the fusion of the two art forms. This collaboration enhances the brand's image and value, provides consumers with new aesthetic experiences, and provides artists with the opportunity to publicize their works. Fashion can also break away from the existing stereotyped patterns and express the zeitgeist contained in modern society in a much more diverse way through collaboration with works of art, which has cultural and social significance. Therefore, the purpose of this study is to rediscover the meaning of fashion in art based on human life and the spiritual world, and to explore how fashion is expressed as a cultural code in modern society. In the past, most of the views looked at the relationship with art as fashion was the subject, but this paper focused on analyzing the interaction between fashion and art from an artistic point of view. This effect could be demonstrated through the analysis of the convergence case of fashion and art. It is expected that various forms of collaboration will continue in the future, especially fashion, which is based on the researcher's abstraction of color, absorbs and inspires each other's elements, and is related in various aspects such as color symbolism, emotional expression, and design composition principles.

Significance of Tissue Expression and Serum Levels of Angiopoietin-like Protein 4 in Breast Cancer Progression: Link to NF-κB /P65 Activity and Pro-Inflammatory Cytokines

  • Shafik, Noha M;Mohamed, Dareen A;Bedder, Asmaa E;El-Gendy, Ahmed M
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8579-8587
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    • 2016
  • Background: The molecular mechanisms linking breast cancer progression and inflammation still remain obscure. The aim of the present study was to investigate the possible association of angiopoeitin like protein 4 (ANGPTL4) and its regulatory factor, hypoxia inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$), with the inflammatory markers nuclear factor kappa B/p65 (NF-${\kappa}B$/P65) and interleukin-1 beta (IL-$1{\beta}$) in order to evaluate their role in inflammation associated breast cancer progression. Materials and Methods: Angiopoietin-like protein 4 (ANGPTL4) mRNA expressions were evaluated using quantitative real time PCR and its protein expression by immunohistochemistry. DNA binding activity of NF-${\kappa}B$/P65 was evaluated by transcription factor binding immunoassay. Serum levels of ANGPTL4, HIF-$1{\alpha}$ and IL-$1{\beta}$ were immunoassayed. Tumor clinico-pathological features were investigated. Results: ANGPTL4 mRNA expressions and serum levels were significantly higher in high grade breast carcinoma ($1.47{\pm}0.31$ and $184.98{\pm}18.18$, respectively) compared to low grade carcinoma ($1.21{\pm}0.32$ and $171.76{\pm}7.58$, respectively) and controls ($0.70{\pm}0.02$ and $65.34{\pm}6.41$, respectively), (p<0.05). Also, ANGPTL4 high/moderate protein expression was positively correlated with tumor clinico-pathological features. In addition, serum levels of HIF-$1{\alpha}$ and IL-$1{\beta}$ as well as NF-${\kappa}B$/P65 DNA binding activity were significantly higher in high grade breast carcinoma ($148.54{\pm}14.20$, $0.79{\pm}0.03$ and $247.13{\pm}44.35$ respectively) than their values in low grade carcinoma ( $139.14{\pm}5.83$, $0.34{\pm}0.02$ and $184.23{\pm}37.75$, respectively) and controls ($33.95{\pm}3.11$, $0.11{\pm}0.02$ and $7.83{\pm}0.92$, respectively), (p<0.001). Conclusion: ANGPTL4 high serum levels and tissue expressions in advanced grade breast cancer, in addition to its positive correlation with tumor clinico-pathological features and HIF-$1{\alpha}$ could highlight its role as one of the signaling factors involved in breast cancer progression. Moreover, novel correlations were found between ANGPTL4 and the inflammatory markers, IL-$1{\beta}$ and NF-${\kappa}B$/p65, in breast cancer, which may emphasize the utility of these markers as potential tools for understanding interactions for axes of carcinogenesis and inflammation contributed for cancer progression. It is thus hoped that the findings reported here would assist in the development of new breast cancer management strategies that would promote patients' quality of life and ultimately improve clinical outcomes. However, large-scale studies are needed to verify these results.

GENE EXPRESSION CHARACTERISTICS OF PUTATIVE PROINFLAMMATORY CYTOKINES AND RECEPTOR MOLECULE CLONING (Putative proinflammatory cytokine유전자의 발현양상과 수용체 분자의 cloing)

  • Oh, Kwi-Ok;Song, Yo-Han;Seo, Young-Seok;Lee, Dong-Whan;Moon, Dae-Hee;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.24 no.3
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    • pp.472-482
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    • 1994
  • Cytokines expressed specifically in leukocytes subsets and in activated cells, which are involved in chemotaxis and activation of leukocytes, are recently defined as chemokines. Macrophage inflammatory $protein-1{\alpha}(MIP-1{\alpha})$ and $MIP-1{\beta}$ are members of C-C chemokine subfamily which produces wide immunomodulatory, proinflammatory, and hematopoietic modulatory actions. We have studied their gene expression by using Northern blot analysis in various blood cells such as cytolytic T lymphocyte(CTL), helper T lymphocyte(HTL), macrophage, and B lymphocyte. Resting CTL line CTLL-R8 expressed $MIP-1{\alpha}$ mRNA which was downregulated by ConA stimulation. Both of resting and ConA stimulated HTL line Hut78 and Jurkat did not express $MIP-1{\alpha}$ mRNA. There was detectable $MIP-1{\alpha}$ transcript in HTL hybridoma 2B4.11 which was a little upstimulated by ConA stimulation. B cell line 230, and macrophage cell line RAW264.7 and WR19M.1 showed distinct $MIP-1{\alpha}$ message which were induced after LPS stimulation. Expression pattern of $MIP-1{\beta}$ in all cell lines or cell were almost identical to that of $MIP-1{\alpha}$. Also strategies employed to identify and characterize the biological functions was preceded by receptor cloning to trace the shorcut to the final goal of cytokine research. For the cloning of $MIP-1{\alpha}$ receptor(R), we used synthetic oligonucleotides of transmembrane(T) conserved sequences of already cloned human(h) IL-8-R, and performed reverse transcription-polymerase chain reaction(RT-PCR) amplification using murine(m) macrophage cell line mRNA. Among 5RT-PCR products, we isolated a homologous cDNA with hIL-8-R which were shown to be putative mIL-8-R cDNA.

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The Relationships among Anger Expression, Interpersonal Relationship, and Depression in Nursing Students (간호대학생의 분노표현 대인관계 및 우울 간의 관계)

  • Byun, Hye-Sun;Kim, Kyung-Hae;Lee, Da-Gyeom
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.4
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    • pp.91-99
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    • 2020
  • This study investigated the relationships of anger expression, interpersonal relationship, and depression in nursing students. The study participants were 209 nursing students recruited from two colleges. The data were collected from questionnaires gathered between December 1st and December 20th, 2016. Data were analyzed by applying descriptive analysis, t-test, ANOVA, Pearson correlation coefficients, and multiple regression analysis, using the SPSS/WIN 19.0 program. Significant correlations were observed for anger expression, interpersonal relationship, and depression in the nursing students. The total explanation of depression was determined to be 39.0%. The anger-in explaining (β=0.506, p=0.000), anger-control explaining (β= -0.342, p=0.000), and interpersonal relationship (β= -0.286, p=0.000) were significant factors of depression. The results of this study indicate a requirement for strategies to decrease trait anger-in, and promote anger-control and interpersonal relationship among nursing students.

The effects of emotional labor of dental hygienist on the job stress, anxiety and sleep (치과위생사의 감정노동이 직무스트레스와 불안, 수면에 미치는 영향)

  • Park, Hye-Young;Jang, Gye-Won;Lee, Gung-Hwa;Lee, Min-Kyung;Lee, Geong-Won;Iim, Yong;Lee, Jong-Ryol;Kim, Hye-Jin
    • Journal of Korean society of Dental Hygiene
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    • v.17 no.3
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    • pp.449-463
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    • 2017
  • Objectives: The objective of this study was to identify the relationship between emotional labor and occupational stress of the dental hygienists, and the effects of emotional labor on their occupational stress and health problems including depression, anxiety and sleep. Methods: The survey was conducted using the questionnaires about dental hygienists' working in medical institutions in Busan, Gyeongsangnam-do from August 19th and October 7th, 2016. Total of 225 participants were selected for data analyses. Frequency analysis, t-test, ANOVA, Pearson's correlation coefficient analysis, and regression analysis were conducted using SPSS Windows ver. 21.0 program(SPSS Inc., Chicago, IL. USA). Results: The analysis of the factors affecting emotional labor, occupational stress, anxiety, and sleep by participants' characteristics showed that those with higher education level, work position and annual salary had higher emotional labor. In addition, participants who worked 5 days every other week and were responsible for patient consultation had higher emotional labor. The analysis of correlations among emotional labor occupational stress, anxiety, and sleep showed positive correlations between emotional experience and emotional expression, occupational stress and emotional expression, and anxiety and emotional expression. On the other hand, negative correlation was found between anxiety and emotional experience. Regression analysis was conducted to examine the effect of emotional labor on job stress. Emotional labor was a factor affecting job stress and anxiety. Conclusions: In order to improve emotional control and vulnerability to stress among dental hygienists, intra-organizational training opportunities and mental health care strategies are needed. Additional broad-based studies are required to identify the factors affecting the occupational stress and emotional labor and to develop relevant intervention measures.

Siamese Crocodile White Blood Cell Extract Inhibits Cell Proliferation and Promotes Autophagy in Multiple Cancer Cell Lines

  • Phosri, Santi;Jangpromma, Nisachon;Chang, Leng Chee;Tan, Ghee T.;Wongwiwatthananukit, Supakit;Maijaroen, Surachai;Anwised, Preeyanan;Payoungkiattikun, Wisarut;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.1007-1021
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    • 2018
  • Cancer represents one of the most significant threats to human health on a global scale. Hence, the development of effective cancer prevention strategies, as well as the discovery of novel therapeutic agents against cancer, is urgently required. In light of this challenge, this research aimed to evaluate the effects of several potent bioactive peptides and proteins contained in crocodile white blood cell extract (cWBC) against LU-1, LNCaP, PC-3, MCF-7, and CaCo-2 cancer cell lines. The results demonstrate that 25, 50, 100, and $200{\mu}g/ml$ cWBC exhibits a strong cytotoxic effect against all investigated cell lines ($IC_{50}$ $70.34-101.0{\mu}g/ml$), while showing no signs of cytotoxicity towards noncancerous Vero and HaCaT cells. Specifically, cWBC treatment caused a significant reduction in the cancerous cells' colony forming ability. A remarkable suppression of cancerous cell migration was observed after treatment with cWBC, indicating potent antimetastatic properties. The mechanism involved in the cancer cell cytotoxicity of cWBC may be related to apoptosis induction, as evidenced by typical apoptotic morphology features. Moreover, certain cWBC concentrations induced significant overproduction of ROS and significantly inhibited the $S-G_2/M$ transition in the cancer cell. The molecular mechanisms of cWBC in apoptosis induction were to decrease Bcl-2 and XIAP expression levels and increase the expression levels of caspase-3, caspase-8, and p53. These led to a decrease in the expression level of the cell cycle-associated gene cyclin-B1 and the arrest of cell population growth. Consequently, these findings demonstrate the prospect of the use of cWBC for cancer therapy.

The Relationship among Narcissism, Usage Motives, and Information Diffusion of Social Media (나르시시즘 성향, 패션소셜미디어 이용동기, 정보확산 행동 간 관계 연구)

  • Kim, Nae-Eun;Song, Gwang-Suk;Kim, Mi-Sook
    • The Journal of Industrial Distribution & Business
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    • v.9 no.1
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    • pp.99-110
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    • 2018
  • Purpose - The purpose of this study is to investigate the relationship among narcissism, usage motives, usage behaviors, satisfaction with and continuance intention to use fashion social media. Research design, data, methodology - A questionnaire survey was used to collect data after conducting a pilot test. Based on the reliability test of the preliminary questionnaire used for the pilot test, the questionnaire was revised. The final questionnaires were administered to 238 fashion social media users and 216 were used for the data analysis. To assess the validity of these measures, exploratory factor analysis and the confirmatory factor analysis were performed. Structural equation modeling analysis were employed for data analysis. Results - Five factors of the usage motivation of fashion social media were extracted: information-seeking, relationship-seeking, practicality-seeking, enjoyment-seeking and self-expression motives. The statistical analysis confirmed the influence of the narcissism tendency on all of the usage motives of fashion social media, three of the fashion social media usage motives influencing information diffusion behavior, and the influence of the information diffusion behavior on users' satisfaction and continuance intention to use fashion social media. Narcissism exerted the highest influences on self-expression motive followed by information-seeking, enjoyment-seeking, relationship-seeking and practicality-seeking motives in order. Factors affecting fashion information diffusion behaviors are practicality-seeking motive, self-expression motive, and relationship-seeking motive. The greater the diffusion of information, the higher the satisfaction with using fashion social media. The consumers with higher satisfaction intended to use fashion social media and share information more frequently. Conclusions - The results indicate that narcissism is an important factor in fashion social media usage motivation. The main motives for narcissistic people to spread information is for the practical purpose at the most, and then to express their personality and style, and to build relationship with others. The satisfaction through active information sharing behaviors seems to play a key role to lead high continuance intention of fashion social media. These implies that marketing strategies to satisfy consumers' narcissism and motives to use social media, and to stimulate the information diffusion behaviors can be used to meet their needs for higher satisfaction with fashion social media.