• Title/Summary/Keyword: quantitative PCR

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De novo Expression of Hepatic UCP3 Is Time-Dependently Related with Metabolic Function in Fenofibrate-Treated High Fat Diet Rats (고지방 섭취한 쥐에서 페노파이브레이트 복용에 의한 간 UCP3 발현 기간과 대사변화 관계)

  • Park, Mi-Kyoung;Kang, Ah-Young;Seo, Eun-Hui;Joe, Yeon-Soo;Kang, Soo-Jeong;Hong, Sook-Hee;Kim, Duk-Kyu;Lee, Hye-Jeong
    • Journal of Life Science
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    • v.19 no.1
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    • pp.15-20
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    • 2009
  • Uncoupling protein 3 (UCP3) is a mitochondrial protein that is expressed predominantly in skeletal muscle. It may play a role in altering metabolic function. However, its major physiological roles are not fully understood. Recently de novo expression of UCP3 in rat liver by fenofibrate was reported. We also reported previously that fenofibrate-induced de novo expression of UCP3 contributes to reduction of adipose tissue in obese rats. In the present study, we investigated that ienofibrate-induced expression of UCP3 in rat liver is related with metabolic function such as body weight and hepatic lipid content by time-dependent manner in high-fat diet rats. Eight-week-old male Sprague-Dawley rats were randomly divided into two groups; the high fat diet group (HF, n=16) and fenofibrate-treated high fat diet group (HFF, n=16). The mRNA expression of hepatic UCP3 was detected as early as 1 week of fenofibrate treatment by quantitative real-time PCR and the amount of mRNA was increased time-dependently. The mean body weight of the HFF group was significantly less com. pared with the HF group after 6 weeks of fenofibrate treatment, even though there was no difference of food intake between the two groups. Rectal temperature was increased during 4 to 6 weeks of fenofibrate treatment and body weight was decreased after 6 weeks of treatment. These results were corresponded with the increased amount of the expression of UCP3 mRNA and protein. We suggest that de novo expression of hepatic UCP3 is increased time-dependently with fenofibrate treatment and that the amount of expression is correlated with metabolic function.

Intracullular Functions of the mas2+ Gene in the Fission Yeast, Schizosaccharomyces pombe (분열형 효모에서의 mas2+ 유전자의 세포 내 기능)

  • Sin, Sang-Min;Cha, Jae-Young;Ha, Se-Eun;Sim, Sun-Mi;Kim, Hyoung-Do;Lee, Jung-Sup;Park, Jong-Kun
    • Journal of Life Science
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    • v.19 no.1
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    • pp.101-110
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    • 2009
  • The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel $mas2^+$ (mitosis associated protein) gene, a homolog of human SMARCAD1 was isolated and characterized from a fission yeast Schizosaccharomyces pombe (S. pombe) using gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 922 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that an SNF2 domain is located, which is involved in the chromosome remodeling. The quantitative analysis of the $mas2^+$ transcript against $adh1^+$ showed that the expression level of $mas2^+$ is high before septum formation in S. pombe. When $mas2^+$ null mutant cells were grown at 27 and $35^{\circ}C$, the cytokinesis of $mas2^+$ null mutant was greatly delayed and a large number of multi-septate and mis-segregated cells were produced. In addition, the number of multi-septate cells significantly increased. When cells were cultured in YES rich medium to increase proliferation, the abnormal phenotypes $mas2^+$ null mutant dramatically increased. These phenotypes could be rescued by an over-expression of the mast gene. The Mas2 protein localized in the nuclei of S. pombe, as evidenced by Mas2-EGFP signals. These results suggest that the $mas2^+$ is homologous to human SMARCAD1 gene and involved in septum formation and chromosome remodeling control.

Data Mining Approaches for DDoS Attack Detection (분산 서비스거부 공격 탐지를 위한 데이터 마이닝 기법)

  • Kim, Mi-Hui;Na, Hyun-Jung;Chae, Ki-Joon;Bang, Hyo-Chan;Na, Jung-Chan
    • Journal of KIISE:Information Networking
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    • v.32 no.3
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    • pp.279-290
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    • 2005
  • Recently, as the serious damage caused by DDoS attacks increases, the rapid detection and the proper response mechanisms are urgent. However, existing security mechanisms do not effectively defend against these attacks, or the defense capability of some mechanisms is only limited to specific DDoS attacks. In this paper, we propose a detection architecture against DDoS attack using data mining technology that can classify the latest types of DDoS attack, and can detect the modification of existing attacks as well as the novel attacks. This architecture consists of a Misuse Detection Module modeling to classify the existing attacks, and an Anomaly Detection Module modeling to detect the novel attacks. And it utilizes the off-line generated models in order to detect the DDoS attack using the real-time traffic. We gathered the NetFlow data generated at an access router of our network in order to model the real network traffic and test it. The NetFlow provides the useful flow-based statistical information without tremendous preprocessing. Also, we mounted the well-known DDoS attack tools to gather the attack traffic. And then, our experimental results show that our approach can provide the outstanding performance against existing attacks, and provide the possibility of detection against the novel attack.

MicroRNA 155 Expression Pattern and its Clinic-pathologic Implication in Human Lung Cancer (폐암에서 microRNA 155의 발현 양상과 임상병리학적 의의)

  • Kim, Mi Kyeong;Moon, Dong Chul;Hyun, Hye Jin;Kim, Jong-Sik;Choi, Tae Jin;Jung, Sang Bong
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1056-1062
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    • 2016
  • Lung cancer is currently the most common malignant disease and the leading cause of mortality in the world and non-small cell lung cancer (NSCLC) accounts for 75-80% of lung cancer cases. miR-155 gene was found to be over expressed in several solid tumors, such as thyroid carcinoma, breast cancer, colon cancer, cervical cancer, pancreatic ductal adenocarcinoma (PDAC) and lung cancer. The aims of this study were to define the expression of miR-155 in lung cancer and its associated clinic-pathologic characteristics. Total RNA was purified from formalin-fixed, paraffin-embedded NSCLC tissues and benign lung tissues. Expression of miR-155 in human lung cancer tissues were evaluated as mean fold changes of miR-155 in cancer tissues compared to benign lung tissues by quantitative real-time reverse transcriptase polymerase chain reaction (real-time qRT-PCR) and associations of miR-155 expression with clinic-pathologic findings of cancer. Compared with the benign control group, miR-155 expression was significantly overexpressed in NSCLCs (p=<0.001). miR-155 was more overexpressed in squamous cell carcinoma than in adenocarcinoma. Poorly differentiated tumors showed significantly overexpression of miR-155 than well-differentiated tumors (p=<0.001). Overexpression of miR-155 was significantly associated with lymph node metastasis (p=<0.05). In survival analysis for all NSCLC patients, high miR-155 expression was significantly correlated with worse overall survival (p=<0.05). These results suggested that miR-155 might play an important role in lung cancer progression and metastasis.

Effect of Hormones and Short Chain Fatty Acids on CYP7A1 Gene Expression in HepG2 Cell (호르몬과 단쇄지방산이 HepG2 Cell 내에서 CYP7A1 발현에 미치는 효과)

  • Yang, Jeong-Lye;Lee, Hyun-Jung;Kim, Yang-Ha
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.5
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    • pp.573-580
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    • 2005
  • Cholesterol $7\alpha-hydroxylase$ (CYP7A1) is the rate-limiting enzyme in the conversion of cholesterol to bile acids and plays a central role in regulating cholesterol homeostasis. We previously showed that a fermentable $\beta-glucan$ ingestion decreased plasma cholesterol levels due to fecal bile acid excretion elevation involved inincrease of cholesterol $7\alpha-hydroxylase$ mRNA expression and activity. It is proposed that short chain fatty acids (SCFA) produced by cecal and colonic fermentation of soluble fiber are associated with cholesterol-lowering effect of fiber. In the present study, we investigated whether CYP7A1 expression is up-regulated by short chain fatty acids or by hormones in cultured human hepatoma (HepG2) cells. Confluent HepG2 cell were incubated with acetate, propionate, or butyrate at 1 mM concentration for 24 hrs. Acetate as well as propionate increased to 1.8-fold expression of CYP7A1 mRNA than the control. Butyrate also increased 1.5-fold expression of CYP7A1 mRNA. Our data show for the first time that SCFA increase expression of CYP7A1 mRNA. Adding insulin, dexamethasone and triiodothyronine $(1\;{\mu}M)$ to HepG2 cell increased the expression of CYP7A1 mRNA to $150\%,\;173\%,\;141\%$, respectively. These results suggest that SCFA produced by cecal fermentation stimulate enteric nervous system, in which secreted some neuropeptides may be responsible for change in cholesterol and bile acid metabolism. These findings suggest that SCFA are involved in lowering plasma cholesterol levels due to the up-regulation of CYP7A1 and bile acid synthesis.

Anti-atopic Effects of Castanea crenata Inner Shell Extracts Fermented by Lactobacillus bifermentans (Lactobacillus bifermentans로 발효한 율피의 항아토피 효과)

  • Kim, Bae Jin;Son, Woo Rim;Choi, Mi Ok;Jo, Seung Kyeung;Jung, Hee Kyoung;Lee, Jin Tae;Kim, Hak Yoon;Kwoen, Dae Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.9
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    • pp.1378-1386
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    • 2013
  • Atopic dermatitis (AD) is a common skin disease characterized by chronic and relapsing inflammatory dermatitis with immunological disturbances. In spite of the continuous increase in the incidence of AD, it is regrettable that till date there is no effective treatment to treat the same. Therefore, the present study was designed to examine the possible anti-atopic effects of Castanea crenata inner shell extracts fermented by Lactobacillus bifermentans (FCS) in 2,4-dinitrochlorobenzene (DNCB) induced AD in NC/Nga mice. Based on the results of HPLC analysis, we found that FCS contains anti-inflammatory factors such as gallic acid (10.18 mg/g) and ellagic acid (2.14 mg/g). The groups that we have used in this study included 0.1%, 1%, 5% fermented Castanea crenata inner shell extracts (FCS 0.1, FCS 1, FCS 5), 1,3-butylene glycol treated control (AD), and normal mice. After topical FCS treatment, we observed that the clinical severity score for AD was lower in both the FCS 1 and FCS 5 groups than the AD group. We also proved beyond doubt that there was improvement of melanin, erythema and skin moisture indices in the FCS 5 group. Spleen index and gene expression levels of pro-inflammatory cytokines such as IL-$1{\beta}$ and TNF-${\alpha}$ were significantly decreased in the FCS 5 group compared to the AD group (P<0.05). Further, we also found that the level of serum immunoglobulin E (IgE) in the FCS-treated group was decreased in a concentration-dependent manner. The results of our study suggest that FCS can be effectively used as a cosmeceutical ingredient for both the prevention and improvement of AD.

Evaluation of Methane Oxidation and the Production Potential of Soils in an Urban School (도심 학교 토양의 메탄 산화 및 생성 잠재력 평가)

  • Lee, Yun-Yeong;Kim, Tae Gwan;Ryu, Hee Wook;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.32-40
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    • 2014
  • Methane oxidation and the production potentials of ground soil (soil A) and garden soil (soil B, C, & D) in an urban school were evaluated, and the methanotrophic and methanogen communities in the soil samples were quantified using quantitative realtime PCR. The methanotrophic community in the raw soil A sample possessed a $6.1{\times}10^3$ gene copy number/g dry weight soil, whereas those in the raw soils B~D samples were $1.6-1.9{\times}10^5$ gene copy numbers/g dry weight soil. Serum bottles added with the soil samples were enriched with methane gas, and then evaluated for their methane oxidation potential. The soil A sample had a longer induction phase for methane oxidation than the other soils. However, soil A showed a similar methane oxidation potential with soils B~D after the induction phase. The methanotrophic community in the enriched soil A sample was increased by up to $2.3{\times}10^7$ gene copy numbers/g dry weight soil, which had no significantly difference compared with those in soils B~D ($1.2-2.8{\times}10^8$ gene copy numbers/g dry weight soil). Methane production showed a similar tendency to methane oxidation. The methanogens community in raw soil A ($1.7{\times}10^5$ gene copy number/g dry weight soil) was much less than those in raw soils B~D ($1.3-3.4{\times}10^7$ gene copy numbers/g dry weight soil). However, after methane gas was produced by adding starch to the soils, soil samples A~D showed $10^7$ gene copy numbers/g dry weight soil in methanogens communities. The results indicate that methanotrophic and methanogenic bacteria have coexisted in this urban school's soils. Moreover, under appropriate conditions for methane oxidation and production, methanotrophic bacteria and methanogens are increased and they have the potential for methane oxidation and production.

Protective effect of lycopene against cytokine-induced β-cell apoptosis in INS-1 cells (라이코펜이 사이토카인에 의해 유도된 베타세포 사멸에 미치는 효과 및 기전 연구)

  • Kim, Kyong;Jang, Se-Eun;Bae, Gong Deuk;Jun, Hee-Sook;Oh, Yoon Sin
    • Journal of Nutrition and Health
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    • v.51 no.6
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    • pp.498-506
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    • 2018
  • Purpose: Lycopene, a carotenoid with anti-oxidant properties, occurs naturally in tomatoes and pink grapefruit. Although the beneficial effects of lycopene on various disorders have been established, little attention has been paid to the possible anti-diabetic effects of lycopene focusing on ${\beta}$-cells. Therefore, this study investigated the potential of lycopene to protect ${\beta}$-cells against apoptosis induced by a cytokine mixture. Methods: For toxicity experiments, the cells were treated with 0.1 ~ 10 nM of lycopene, and the cell viability in INS-1 cells (a rat ${\beta}$-cell line) was measured using a MTT assay. To induce cytokine toxicity, the cells were treated with a cytokine mixture (20 ng/mL of $TNF{\alpha}$ + 20 ng/mL of IL-$1{\beta}$) for 24 h, and the effects of lycopene (0.1 nM) on the cytokine toxicity were measured using the MTT assay. The expression levels of the apoptotic proteins were analyzed by Western blotting, and the level of intracellular reactive oxidative stress (ROS) was monitored using a DCFDA fluorescent probe. The intracellular ATP levels were determined using a luminescence kit, and mRNA expression of the genes coding for anti-oxidative stress response and mitochondrial function were analyzed by quantitative reverse-transcriptase PCR. Results: Exposure of INS-1 cells to 0.1 nM of lycopene increased the cell viability significantly, and protected the cells from cytokine-induced death. Lycopene upregulated the mRNA and protein expression of B-cell lymphoma-2 (Bcl-2) and reduced the expression of the Bcl-2 associated X (Bax) protein. Lycopene inhibited apoptotic signaling via a reduction of the ROS, and this effect correlated with the upregulation of anti-oxidative stress response genes, such as GCLC, NQO1, and HO-1. Lycopene increased the mRNA expression of mitochondrial function-related genes and increased the cellular ATP level. Conclusion: These results suggest that lycopene reduces the level of oxidative stress and improves the mitochondrial function, contributing to the prevention of cytokine-induced ${\beta}$-cell apoptosis. Therefore, lycopene could potentially serve as a preventive and therapeutic agent for the treatment of type 2 diabetes.

Moderate Intensity Exercise Has More Positive Effects on The Gene Expression of Inflammasome, M1, M2 Macrophage Infiltration and Brown Adipocyte Markers Compared to High Intensity Exercise in Subcutaneous Adipose of Obese Mice Induced By High Fat Diet (비만모델에서 중강도 운동에 의한 인플라마좀, 대식세포 침윤, 갈색지방 관련 바이오 마커의 개선 효과)

  • Kim, Yong-An;Pitriani, Pipit;Park, Hee-Geun;Lee, Wang-Lok
    • Journal of Life Science
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    • v.29 no.3
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    • pp.303-310
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    • 2019
  • The purpose of the study was to compare the effect of either moderate or high intensity aerobic exercise on inflammasome, M1, M2 macrophage infiltration and brown adipocyte markers in subcutaneous adipose tissue of the high fat diet-induced obese mice. The 4 weeks male C57BL/6 mice were randomly assigned to four groups: normal diet control (NC; n=10), high-fat diet control (HC; n=10), high fat diet with moderate intensity exercise (HME; n=10), or high fat diet with high intensity exercise (HIE; n=10) groups. The high fat diet was given 60% calories from fat whereas normal diet was given 18% calories from fat. The moderate intensity exercise group (HME) was set at 10m/min in the first 2 weeks, 12m/min in 3-5 weeks and 14m/min in 6-16 weeks and the high intensity exercise group (HIE) was set at 14m/min in the first 2 weeks, 17m/min in 3-5 weeks and 18m/min in 6-16 weeks. The semi quantitative reverse transcription-polymerase chain reaction (RT PCR) was used to analyze the gene expression. The moderate intensity exercise significantly reduced the expression of NLRP3, F480, CD11c and CD86. Further, the moderate intensity exercise significantly increased CD206 and $PGC1{\alpha}$, BMP7 and PRDM. The high intensity exercise significantly reduced NLRP3, CD11c and CD86. Further, the high intensity exercise significantly increased $PGC1{\alpha}$ and BMP7. In conclusion, moderate intensity exercise has more positive effects on inflammasome, M1, M2 macrophage infiltration and brown adipocyte maskers compared to high intensity exercise in high fat diet induced obese mice.

Diagnostic Availability of Estrogen Receptor Alpha mRNA on Cervical Cancer Tissue (자궁경부암 조직에서 에스트로겐 수용체 알파 mRNA의 진단적 유용성)

  • Kim, Geehyuk;Yu, Kwangmin;Kim, Jungho;Kim, Seoyong;Park, Sunyoung;Ahn, Sungwoo;Lee, Ji-Young;Kim, Sunghyun;Park, Ho-Hyun;Lee, Dongsup
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.449-456
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    • 2018
  • Cervical cancer is the fourth most frequently diagnosed cancer in women worldwide. In lower Human Development Index countries, it has the second highest incidence and mortality among cancer in women. Therefore, better diagnosis and treatment systems are needed. Among them, estrogen receptor alpha ($ER-{\alpha}$) mRNA expression has been analyzed with RT-qPCR since several studies reported that $ER-{\alpha}$ is necessary in the maturation of the uterus and is related to cervical cancer. In this study, $ER-{\alpha}$ quantitative analysis was performed on various lesions and normal tissue samples. Based on the receiver operating characteristic (ROC) curve, its sensitivity and specificity were 85% and 75%, respectively, showing higher or similar results to those of conventional HPV tests. In addition, its expression level was analyzed with clinical information. With regression analysis, the R square value between the $ER-{\alpha}$ mRNA expression level and menopause status was 0.5041, indicating a strong correlation. This study was performed as part of a pilot study and suggests that $ER-{\alpha}$ is related to carcinogenesis. Future studies will examine other hormones and menopausal factors with a larger sample size.