• Title/Summary/Keyword: 유전자독성

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Chitosan Derivatives for Target of Specific Tissue in the Body (생체 내 특정 조직의 표적을 위한 키토산 유도체)

  • Jang, Mi-Kyeong;Nah, Jae-Woon
    • Applied Chemistry for Engineering
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    • v.21 no.6
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    • pp.593-602
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    • 2010
  • Chitosan as a natural polymer has superior physicochemical properties such as biocompatibility, biodegradability and nontoxicity, but application of chitosan for therapy of cancer and gene related-disease has been limited by poor solubility in aqueous solution. Therefore, low molecular weight water-soluble chitosan (LMWSC) with high reactivity and strong positive charge can be applied as a delivery system having function to carry in the specific tissue the bioactive material like poor solubility drug, or therapeutic gene and developed as a therapeutic system having good therapeutic efficiency. The most important factor for therapy of various diseases is to reveal the antigen or receptor expressed in specific lesion tissue and the antibody and ligand which can bind with antigen is to introduce at the biomaterials for enhancement the therapeutic efficiency. The studies for cationic synthetic polymer as drug or gene delivery have been actively performed, but it has many problems such as toxicity in the body, therapeutic efficiency. From this point of view, this article demonstrated the introduction of functional groups to target the specific tissue and therapeutic strategy using the modification of LMWSC with free-amine group. The development of these delivery system will provide a positive vision for cancer therapy.

Effects of Poly-Gamma Glutamate Contents Cheonggukjang on Osteoblast Differentiation (폴리감마글루탐산(PGA) 함유량이 증가된 청국장이 조골세포 분화에 미치는 영향)

  • Lee, Ki Ho;Sim, Mi-Ok;Song, Yong Su;Jung, Ho Kyung;Jang, Ji-Hun;Kim, Min-Suk;Kim, Tae Mook;Lee, Hyo Eun;An, Byeong-Kwan;Jung, Won Seok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.5
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    • pp.664-670
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    • 2016
  • Cheonggukjang (CKJ) is a Korean traditional food made of fermented soybeans. In comparison to normal intake of soybeans, Cheonggukjang has high digestibility with bioactive, antioxidant substances, and thrombolytic enzymes. Recent studies have reported anti-oxidant, anti-cancer, anti-inflammatory, anti-obesity activities as well as inhibitory activities against osteoporosis for CKJ. In this study, we identified the effects of CKJ on osteoblast differentiation by increasing the polyglutamic acid (PGA) content of CKJ. Alkaline phosphatase (ALP) activity and mineralization significantly increased in response to treatment with both natural CKJ (CKJ A) and PGA-increased CKJ (CKJ B). However, CKJ B exhibited higher ALP activity and mineralization than CKJ A. Real-time reverse transcription PCR demonstrated that mRNA expression of osteoblastic-associated genes such as type I collagen, alkaline phosphatase, osteocalcin, and osteopontin in C2C12 cells was significantly up-regulated by CKJ A or B treatment. These results indicate that treatment with CKJ has an anabolic effect on bone by increasing osteoblastic differentiation and ALP activity. Increasing PGA content in CKJ had a greater effect than CKJ A on up-regulation of osteoblastic gene expression in osteoblast cells.

Inhibitory effect of Allium macrostemon extracts on adipogenesis of 3T3-L1 preadipocytes (산달래 추출물의 3T3-L1 지방전구세포 분화 억제 효능)

  • Lee, Joo-Yeon;Jeong, Yeju;Kim, Jina;Kim, Choon Young
    • Korean Journal of Food Science and Technology
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    • v.52 no.5
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    • pp.441-449
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    • 2020
  • The aim of this study was to compare the biological activities of whole-plant (WAE), bulb (BAE), and leaf (LAE) extracts of Allium macrostemon. The antioxidant activities, total polyphenol contents, and anti-adipogenic activities of WAE and LAE were superior to those of BAE, whereas the biological effects of WAE and LAE were similar. Therefore, the effect of LAE on adipogenesis was further investigated. Treatment of preadipocytes with LAE at 100 g/mL resulted in the inhibition of intracellular lipid accumulation by 49.64%. Consistent with this result, quantitative reverse transcription-PCR showed that LAE treatment decreased the gene expressions of CCAAT/enhancer-binding protein beta (C/EBPβ), peroxisome proliferator-activated receptor gamma (PPARγ), C/EBPα and stearoyl-CoA desaturase 1 (SCD1). Thus, LAE attenuates the adipogenesis of preadipocytes by suppressing the expression of adipogenic and lipogenic genes. These results suggest that LAE can be potentially useful as a functional ingredient to prevent obesity in the food industry.

Expression of Human Cytochrome b5 in Zebrafish (Zebrafish에서 human cytochrome b5의 발현)

  • Han, Se Mi;Yoo, Min
    • Journal of Life Science
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    • v.27 no.6
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    • pp.617-622
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    • 2017
  • In this study, we sought to develop an effective cloning system by which human cytochrome $b_5$ (cyt $b_5$) is introduced and expressed in zebrafish. First, the 414 bp human cyt $b_5$ gene was amplified from RNA extracts of HeLa cells using RT-PCR, and the amplicon was subsequently sequenced to confirm that it was intact. Next, cyt $b_5$ was cloned into the pEGFP-N3 vector, which also encodes a fluorescent gene. One-cell stage zebrafish embryos were microinjected with the recombinant vector containing the cyt $b_5$ gene. Fluorescence microscopy confirmed high expression of the fluorescent gene in the injected fry compared to the non-fluorescent control fry. Finally, we extracted RNA from the injected fry and performed RT-PCR to determine whether the human cyt $b_5$ gene is expressed in the transgenic zebrafish. Sequencing analysis further confirmed that the cloned human cyt $b_5$ gene was intact. The transgenic zebrafish model produced in this study will be a useful tool to study therapeutic approaches to cure various diseases related to the deficiency of functional human cyt $b_5$ as well as tools for cloning useful genes in fish.

Status and Prospect of Lactic Acid Bacteria with Antibiotic Resistance (항생제 내성을 가진 유산균의 현황과 전망)

  • Chon, Jung-Whan;Seo, Kun-Ho;Bae, Dongryeoul;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • v.38 no.2
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    • pp.70-88
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    • 2020
  • Lactic acid bacteria (LAB) form an essential part of the intestinal microbiota of the human body and possess the ability to stabilize the intestinal microbiota, strengthen immunity, and promote digestion as well as intestinal synthesis of vitamins, amino acids, and proteins. Hence, LAB are currently widely used in various products. However, due to the indiscriminate overuse of antibiotics in humans and livestock, bacterial resistance to antibiotics has been increasing rapidly, which has led to serious problems in the treatment of bacterial infections. Additionally, several reports have revealed that antibiotic-resistant LAB may infect people whose immune systems are not fully developed or whose immune systems are temporarily weakened. Therefore, it is imperative to consider the possibility of antibiotic-resistant LAB causing diseases in humans and animals, investigate the mechanism of action between antibiotics and LAB, and determine the relevant regulations for the safe use of LAB.

Gene Expression of Chironomus riparius Heat Shock Protein 70 and Developmental Retardation Exposure to Salinity (염분 노출에 따른 Chironomus riparius의 성장지연과 Heat shock protein 70 유전자 발현)

  • Kim, Won-Seok;Im, Byeong-Hyeon;Hong, Cheol;Choi, Seung-Won;Park, Kiyun;Kwak, Ihn-Sil
    • Korean Journal of Ecology and Environment
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    • v.50 no.3
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    • pp.305-313
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    • 2017
  • We investigate that the impact of freshwater organism exposed to the salinity environment by the frequent rainfall following climate change. To evaluate the stress response following salinity exposure, we assessed the survival rate, molting success rate, the developmental period and mouthpart deformities in Chironomus riparius. In addition, we measured the molecular responses of biomarker gene, gene expression of heat shock protein 70 (HSP70) in C. riparius exposed to salinity after 96 hour. The C. riparius survival rates were showed on time dependent manner and not observed survival organisms above 15 psu at day 4. The pupation and emergence of C. riparius were not seen above 15 psu, and the molting success rate was less than 20% at 10 psu. The developmental retardation of C. riparius was well observed in the pupation and emergence period and was delayed by 4 days at 10 psu compared to the control and 5 psu. The mouthpart deformities after salinity exposure at 96 or 72 hour were observed at 10 psu and 15 psu. The expression of C. riparius HSP70 level was significantly increased exposure to 5 psu and 10 psu. Thus, salinity has been caused to be various ecotoxicological and molecular stress responses on freshwater organisms similar to harmful substances such as EDCs and so on.

Characteristics of Growth and Metal Removal in Recombinant Saccharomyces cerevisiae harboring a Metallothionein Gene (Metallothionein 유전자가 도입된 재조합 Saccharomyces cerevisiae의 생육과 금속제거에 대한 특성)

  • 정동환;김대옥서진호
    • KSBB Journal
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    • v.10 no.5
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    • pp.475-481
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    • 1995
  • The effect of metallothionein expression on the metal resistance and removal by recombinant Saccharomyces cerevisiae containing the plasmid pJW9 was investigated. The recombinant strain S. cerevisiae BZ-pJ was constructed by transforming the host strain S. cerevisiae BZ3l-1-7Ba with the gene coding for a metal-binding protein, metallothionein. Introduction of the MT gene yielded an increase in the minimum inhibitory concentration (MIC) of copper more than three times compared with the host strain. The minimum inhibitory concentrations of $Cr^{2+}, Znr^{2+} and Pb^{2+}, $ were not different for the two strains. The recombinant yeast grown in a medium containing 8mM CuSO4 was able to remove copper with a capacity of 18.9mg $Cu^{2+}$/g dry cell. In a mixture of copper and zinc, the presence of copper relieved the toxic effects caused by zinc, resulting in an enhancement of the final cell density and the specific growth rate of the recombinant yeast. The capability to remove copper by the recombinant yeast was linearly proportional to the copper concentrations in the medium. The efficiency of copper removal was rather constant regardless of the initial copper concentrations. The specific removal of zinc was dependent on the zinc concentrations in media, though, and such dependence was not so pronounced as the concentration of copper.

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The Workflow for Computational Analysis of Single-cell RNA-sequencing Data (단일 세포 RNA 시퀀싱 데이터에 대한 컴퓨터 분석의 작업과정)

  • Sung-Hun WOO;Byung Chul JUNG
    • Korean Journal of Clinical Laboratory Science
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    • v.56 no.1
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    • pp.10-20
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    • 2024
  • RNA-sequencing (RNA-seq) is a technique used for providing global patterns of transcriptomes in samples. However, it can only provide the average gene expression across cells and does not address the heterogeneity within the samples. The advances in single-cell RNA sequencing (scRNA-seq) technology have revolutionized our understanding of heterogeneity and the dynamics of gene expression at the single-cell level. For example, scRNA-seq allows us to identify the cell types in complex tissues, which can provide information regarding the alteration of the cell population by perturbations, such as genetic modification. Since its initial introduction, scRNA-seq has rapidly become popular, leading to the development of a huge number of bioinformatic tools. However, the analysis of the big dataset generated from scRNA-seq requires a general understanding of the preprocessing of the dataset and a variety of analytical techniques. Here, we present an overview of the workflow involved in analyzing the scRNA-seq dataset. First, we describe the preprocessing of the dataset, including quality control, normalization, and dimensionality reduction. Then, we introduce the downstream analysis provided with the most commonly used computational packages. This review aims to provide a workflow guideline for new researchers interested in this field.

Genotoxicological Safety of Hot Water Extracts of the ${\gamma}$-Irradiated Glycyrrhizae Radix, Aurantii nobilis Pericarpium and Bupleuri Radix in vitro (감마선조사 감초, 진피(陳皮) 및 시호 열수 추출물의 in vitro 유전독성학적 안전성 평가)

  • Jo, Sung-Kee;Ham, Yeon-Ho;Park, Hae-Ran;Oh, Heon;Byun, Myung-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1137.2-1245
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    • 2001
  • The ${\gamma}$-irradiated medicinal herbs were examined the genotoxicological safety to consider the possibility of application of the irradiation technology for hygienic purpose. The three medicinal herbs -Glycyrrhigae Radix, Aurantii nobilis Pericarpium and Bupleuri Radix- were irradiated with ${\gamma}$ -rays at the practical dosage of 10 kGy. The hot water extracts of the irradiated herbs were examined in two short-term in vitro tests: (1) Ames test in Salmonella typhimurium TA98 and TA100, (2) Micronucleus test in cultured Chinese hamster ovary(CHO) cells. In the Salmonella reversion assays both with and without metabolic activation, the number of revertant colonies was not increased with each extract from the irradiated herbs, compared with negative controls. No significant difference in formation of the colonies was observed between non-irradiated and 10 kGy-irradiated herbs. These results indicated that no mutagenicity of the irradiated herbs was detected. In the micronucleus tests in cultured CHO cells, the incidences of micronucleus were not increased with irradiated herbs, and no significant difference in the incidences was observed between non-irradiated and irradiated herbs. These results indicated that no cytogenetic toxicity of the irradiated herbs was detected. The results of the two in vitro tests suggest that the irradiated herbs do not show mutagenic effects and cytogenetic toxicity. Further tests of in vivo genotoxicity and chronic toxicity are needed to determine the safety of the herbs irradiated with ${\gamma}$ -rays at practical doses.

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Mutant Frequency at the hprt Locus in Human T-Cell Exposed to Pentachlorophenol (Pentachlorophenol의 노출에 의한 사람 T-임파구의 hprt 유전자에서 돌연변이 빈도)

  • 윤병수;조명행;김인규;박선영;이영순
    • Toxicological Research
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    • v.13 no.1_2
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    • pp.71-78
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    • 1997
  • The mutational effects of pentachlorophenol (PCP) on the hypoxanthine phosphoribosyl transf erase (hprt) locus in human T-cell were analysed by T-cell clonal assay in vitro. Cells were exposed for 24 hours at primary culture to 0~100 ppm (W/V) PCP in dimethyl sulfoxide. Treated cells were allowed at the same time to stimulate by phytohemagglutinin (PHA) and T-cell growth factor (TCGF) and then seeded in medium containing 6-thioguanine to select for hprt-negative routants. We have also defined the optimal condition for the determination of mutant frequency. The parameters investigated include survival counting, first and second subculture for clonal efficiency plating and mutant plating. Under the optimal conditions, mutant frequencies of high dose-treated cells were significantly higher than those of non-treated or low dose cells. The results indicated a clear dose-effect relationship and showed that mutant frequency in 50 ppm PCP treated cell was 4.31$\times$$10^{-5}$ (background, 8.32$\times$$10^{-6}$). Above data strongly suggest that hprt mutation assay can be used as a biomarker for the environmental risk assessment.

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