• Title/Summary/Keyword: CAT mRNA

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The Effect of Stocking Density and Strain on the Performance and Physiological Adaptive Responses in Broiler Chickens (육계에서 품종 및 사육 밀도가 생산성 및 생리적 적응성 지표에 미치는 영향)

  • Jang, In-Surk;Yun, Seo-Hyun;Ko, Young-Hyun;Kim, Se-Yun;Song, Min-Hye;Kim, Jong-Sun;Sohn, Sea-Hwan;Moon, Yang-Soo
    • Korean Journal of Poultry Science
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    • v.41 no.3
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    • pp.205-215
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    • 2014
  • The aim of this study was to determine the effects of stocking density and strain on the performance and physiological adaptive responses including the plasma corticosterone content and the level of mRNA expression of pro-inflammatory cytokines and antioxidant enzymes in broiler chicks. A total of 300 birds of two strains (150 Ross strain vs. 150 Cobb strain) aged 3-d old were allotted into two stocking densities (standard stocking density,$0.046m^2/bird$ vs. high stocking density, $0.023m^2/bird$) in battery cages by $2{\times}2$ factorial designs with ten replicates until 35 d of age. There was no significant strain effect on body weight, feed intakes and feed to gain ratio and the relative organ weights. However body weight, feed intakes and relative organ weight were found to be significantly (P<0.05) affected by the effect of stocking density. Plasma corticosterone level was not affected by both stocking density and strain effects. Hepatic mRNA expression of pro-inflammatory cytokines including interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, IL-18 and interferon-gamma (IFN-${\gamma}$) was not significantly changed by the effects of strain and stocking density. However, the mRNA expression of glutathione peroxidase (GPX) was affected by strain, showing that Ross strain decreased (P<0.05) the GPX expression. With respect to the effect of stocking density, there was a significant (P<0.05) increase in superoxide dismutase (SOD) and catalase (CAT) and GPX mRNA expression in the liver from high stocking density group. Splenic pro-inflammatory cytokine expression was not also affected by stocking density and strain, except that IL-18 mRNA significantly (P<0.05) decreased in Cobb strain under high stocking density. The mRNA expression of SOD and CAT was significantly (P<0.05) affected by the effects of stocking density and strain. In conclusion, growth performance was not affected by strain but stocking density. Although mRNA expression of major pro-inflammatory cytokines was not changed by stocking density and strain, antioxidant enzyme was significantly affected by stocking density, strain or even organ in birds under summer conditions. More detailed studies still needed to be explored to elucidate the effects of environmental conditions and genetic background on physiological responses in birds.

Anti-aging Effect of Cycloheterophyllin in UVA-irradiated Dermal Fibroblasts (자외선 조사에 의해 노화된 섬유아세포에서 Cycloheterophyllin의 항노화 효능)

  • Shim, Joong Hyun
    • Korean Journal of Pharmacognosy
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    • v.50 no.4
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    • pp.285-290
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    • 2019
  • This study was carried out to identify the skin anti-aging effect of cycloheterophyllin on dermal fibroblasts. To elucidate anti-aging effects of cycloheterophyllin on dermal fibroblasts, I measured cell viability, mRNA expressions, and Collagen, type I/matrix metallopeptidase 1(MMP1)-ELISA assay. In this study, I investigated the effects of cycloheterophyllin on Collagen, type I, alpha 1(COL1A1)/Collagen, type III, alpha 1(COL3A1)/MMP1/Superoxide dismutases/Catalase(CAT) mRNA expressions and Collagen, type I/MMP1 protein production. Quantitative Real-time RT-PCR showed that cycloheterophyllin increased mRNA level of COL1A1/COL3A1/CAT genes and collagen, type I protein by ELISA assay compared to UVA-treated dermal fibroblasts. Furthermore MMP1 mRNA and protein expressions were decreased by cycloheterophyllin treatment. These observations revealed that cycloheterophyllin increased anti-aging effects in dermal fibroblasts. Therefore, I identified the anti-aging effects of cycloheterophyllin, and these results showed that the cycloheterophyllin can be a considerable potent ingredient for skin anti-aging. Based on this, I anticipated further researches about cycloheterophyllin for mechanism to develop not only cosmetics but for healthcare food or medicine.

Transcriptional Response of Major Antioxidant Enzyme Genes to Heat Stress in Mud Loach (Misgurnus mizolepis) (고온 스트레스에 대한 미꾸라지(Misgurnus mizolepis) 항산화 효소 유전자들의 발현 특징)

  • Cho Young-Sun;Lee Sang-Yoon;Bang In-Chul;Kim Dong-Soo;Nam Yoon-Kwon
    • Journal of Aquaculture
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    • v.19 no.3
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    • pp.157-165
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    • 2006
  • Expression of major antioxidant enzyme (AOE) including Cu/Zn superoxide dismutase (Cu/Zn-SOD), catalase (CAT), glutathione-S-transferase (GST) and 3 glutathione peroxidase isotypes (GPXs) at mRNA levels during heat stress was examined in mud loach (Misgurnus mizolepis) liver. Based on the semi-quantitative RT-PCR, real-time RT-PCR and/or northern dot blot hybridization, the antioxidant enzyme genes were generally up-regulated during elevation of water temperature from $23^{\circ}C$ up to $32^{\circ}C$. GPXs and SOD displayed the most significant elevation of mRNA levels (up to 3 and 2 folds, respectively) while CAT showed the steady-state expression irrespective of thermal conditions. GST represented the relatively moderate response (1.3-fold increase) in its transcription to thermal stress. The transcriptional activation of AOE genes was not significant at the treatment temperature lower than $29^{\circ}C$. Increased mRNA levels of GPX (extracellular form) and SOD genes in the fish exposed to $32^{\circ}C$ was readily detectable 1 day after exposure to heat stress.

Oxidative Stress and Apoptosis in Goldfish (Carassius auratus) Caused by Exposure to Different Concentrations of Micro-polystyrene

  • Li, Zhongze;Song, Jin Ah;Choi, Cheol Young
    • Ocean and Polar Research
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    • v.43 no.3
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    • pp.141-148
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    • 2021
  • Microplastic contamination in waterbodies is a growing source of concern for researchers and other stakeholders. We investigated oxidative stress and toxicity in goldfish (Carassius auratus) in response to exposure to 1-㎛ diameter micro-polystyrene (MP) at concentrations of 0, 10, 100, and 1000 beads/mL (MP 0, MP 10, MP 100, and MP 1000 groups) for 7 d (at day 0, 1, 3, 5, and 7). We analyzed the survival rates; superoxide dismutase (SOD) and catalase (CAT) mRNA expression levels in the liver; SOD and CAT activity in the plasma; caspase-3 mRNA expression in the liver; and the levels of hydrogen peroxide (H2O2) in plasma. Terminal transferase dUTP nick end labeling (TUNEL) assays were also conducted to determine apoptosis levels in the liver. All fish in the MP 1000 group died by day 7 and the MP 100 group had a lower survival rate than the MP 10 and MP 0 groups. The mRNA expression as well as SOD, CAT, and caspase-3 activity levels were increased significantly with increases in MP concentration and exposure time. Finally, according to the TUNEL assay, more apoptosis was observed in the MP 1000 group at day 5 than in other groups. In summary, MP concentrations above 100 beads/mL caused death and oxidative stress to goldfish. We conclude that MP can cause oxidative stress and apoptosis in goldfish, which leads to death.

Physiological Changes of Juvenile Abalone, Haliotis sieboldii Exposed to Acute Water-temperature Stress (급격한 수온 스트레스에 따른 시볼트전복, Haliotis sieboldii 치패의 생리적 변화)

  • Kim Tae-Hyung;Kim Kyung-Ju;Choe Mi-Kyung;Yeo In-Kyu
    • Journal of Aquaculture
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    • v.19 no.2
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    • pp.77-83
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    • 2006
  • This study was conducted to investigate changes of hemolymph count, antioxidant enzyme activities (catalase: CAT and superoxide dismutase: SOD) and Heat Shock Protein 70 (HSP70) mRNA in hemolymph, hepatopancreas and gill of abalone (Haliotis sieboldii) exposed to various water temperatures. Abalones were exposed to 10, 15, 20, 25 or $30^{\circ}C$ for 0, 6, 12, 24 or 48 hours. Survival rate of abalone was 100% at 10, 15, 20 and $25^{\circ}C$, but 0% at $30^{\circ}C$. Hemolymph counts increased at lower water temperatures (10 and $15^{\circ}C$) and decreased at $30^{\circ}C$. SOD activity decreased immediately after exposure to lower or higher water temperatures compared to the control ($20^{\circ}C$) with an exception at $30^{\circ}C$ where the activity increased. At lower temperatures, SOD activity rose high after 24 hours, but decreased again at 48 hours. At $25^{\circ}C$, it decreased compared to the control. CAT activity decreased immediately after exposure to 10 or $25^{\circ}C$ compared to the control, and then was recovered to the initial level after increment. At $15^{\circ}C$, CAT activity was high after 6 hours, and then was recovered to the initial level after increment. At $30^{\circ}C$, the activity decreased throughout the experiment. The HSP70 mRNA expression in gill increased at lower temperatures compared to the control ($20^{\circ}C$) and $25^{\circ}C$. In this study, rapid change of wale, temperature caused stress response in abalone which had been raised at $20^{\circ}C$. At molecular level, HSP70 was expressed rapidly, but antioxidant enzymes like SOD and CAT were expressed later than HSP70. At 15 and $25^{\circ}C$ of water temperatures, the HSP70, SOD and CAT expression were stable with time. However, at $30^{\circ}C$, all abalone died possibly because they could not develop resistance to high temperature.

Prevalence of Feline Hemotropic Mycoplasmas Among Feral Cats in Korea by use of a PCR assay

  • Cho, Eun Kwang;Yu, DoHyeon;Choi, Ul Soo
    • Journal of Veterinary Clinics
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    • v.33 no.3
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    • pp.145-150
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    • 2016
  • We determined the prevalence of feline hemotropic mycoplasma species including 'Candidatus Mycoplasma haemominutum', Mycoplasma haemofelis, and 'Candidatus Mycoplasma turicensis' in naturally infected feral cats in Jeonju, Korea. Forty six feral cats were evaluated by PCR assay targeting the 16S rRNA gene sequence. Nine cats (19.6%) were positive for 'Candidatus Mycoplasma haemominutum', 2 cats (4.3%) were positive for 'Mycoplasm a haemofelis', and 1 cat (2.2%) was infected with both 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemofelis. 'Candidatus Mycoplasma turicensis' was undetected. Partial 16S rRNA gene sequences of Mycoplasma haemofelis were closely (> 96%) related to those from other countries. The amplification of hemoplasma DNA in these samples confirmed the presence of 'Candidatus M. haemominutum' and M. haemofelis in Korea.

Mycobacterium tuberculosis ESAT6 and CPF10 Induce Adenosine Deaminase 2 mRNA Expression in Monocyte-Derived Macrophages

  • Bae, Mi Jung;Ryu, Suyeon;Kim, Ha-Jeong;Cha, Seung Ick;Kim, Chang Ho;Lee, Jaehee
    • Tuberculosis and Respiratory Diseases
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    • v.80 no.1
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    • pp.77-82
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    • 2017
  • Background: Delayed hypersensitivity plays a large role in the pathogenesis of tuberculous pleural effusion (TPE). Macrophages infected with live Mycobacterium tuberculosis (MTB) increase the levels of adenosine deaminase2 (ADA2) in the pleural fluid of TPE patients. However, it is as yet unclear whether ADA2 can be produced by macrophages when challenged with MTB antigens alone. This study therefore evaluated the levels of ADA2 mRNA expression, using monocyte-derived macrophages (MDMs) stimulated with MTB antigens. Methods: Purified monocytes from the peripheral blood mononuclear cells of healthy volunteers were differentiated into macrophages using granulocyte-macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor (M-CSF). The MDMs were stimulated with early secretory antigenic target protein 6 (ESAT6) and culture filtrate protein 10 (CFP10). The mRNA expression levels for the cat eye syndrome chromosome region, candidate 1 (CECR1) gene encoding ADA2 were then measured. Results: CECR1 mRNA expression levels were significantly higher in MDMs stimulated with ESAT6 and CFP10, than in the unstimulated MDMs. When stimulated with ESAT6, M-CSF-treated MDMs showed more pronounced CECR1 mRNA expression than GM-CSF-treated MDMs. Interferon-${\gamma}$ decreased the ESAT6- and CFP10-induced CECR1 mRNA expression in MDMs. CECR1 mRNA expression levels were positively correlated with mRNA expression of tumor necrosis factor ${\alpha}$ and interleukin 10, respectively. Conclusion: ADA2 mRNA expression increased when MDMs were stimulated with MTB antigens alone. This partly indicates that pleural fluid ADA levels could increase in patients with culture-negative TPE. Our results may be helpful in improving the understanding of TPE pathogenesis.

IN HUMAN BREAST CANCER MCF-7 CELLS, ESTROGEN INVOLVES IN CYPIA1 GENE EXPRESSION.

  • Hwang, J.E.;S.H.Eo;Cho, S.N.;Y.Y.Sheen
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.107-107
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    • 1997
  • Cytochrome P450 enzymes have been intensively investigated in hepatic tissues and several mammalian cell lines. Compared to most studies about cytochrome P450 isozymes in liver in vivo and hepatic, cell lines in vitro, the study of cytochrome P450IA1 in human breast cancer cells could be very important to understand the mechanism of the regulation of CYPIA1 gene expression and cell growth. MCF-7 human breast cancer cells are well characterized to study estrogen and antiestrogen action due to the fact that they contain high level of estrogen receptor and have biological markers characterized. And also MCF-7 cells express high level of arylhydrocarbon hydroxylase activity and human cytochrome P450IA1 cDNA was cloned from MCF-7 cells. Ah receptor was characterized in many breast cancer cell lines and polycyclic aromatic hydrocarbon such as 3-MC induced the expression of CYPIA1 gene and cytochrome P450- dependent monooxygenase activity. We undertook a study to examine the effect of estrogens and other chemicals on the regulation of human CYPIA1 gene expression in MCF-7 cells via RTPCR analysis, that might help us to understand the mechanism of the regulation of CYPIA1 gene expression and MCF-7 cell growth. Expression vector containing the functional 5'-regulatory region of human CYPIA1 fused to the CAT reporter gene was transfected into estrogen receptor positive MCF-T cells or estrogen receptor negative MDA-MB-231 cells. After these cells were treated with various chemicals, RTPCR was carried out to measure both CYPIA1 mRNA and CAT mRNA levels. 1nM 3-MC increased in both P450 and CAT mRNA levels over those of control by two folds in MCF-7 cells but does not in MDA-MB-231 cells. Estrogen or tamoxifen or retinoic acid or chrysin decreased in both P450 and CAT mRNA levels that were induced by 3-MC in MCF-7 when each chemical was administered with 3-MC concomitantly. These results suggested that the level of CYPIA1 gene expression is modulated with estrogen-related molecules and make it possible to speculate that ER is related to CYPIA1 gene expression and cell growth in breast cancer cells. [Supported by grants from the Korean Ministry of Education ]

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Expression of the Genes Involved in the Synthesis of Riboflavin from Photobacterium species of Bioluminescent Marine Bacteria (해양 발광 박테리아 Photobacterium Species의 Riboflavin 생합성에 관여하는 유전자들의 발현)

  • 이찬용
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.1-7
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    • 2000
  • The genes involved in riboflavin synthesis (ribI, II, III, and IV) were found immediately downstream of luxG in the lux operon from Photobacterium species. The single stranded DNA containing the intergenic region of lux genes and rib genes from Photobacterium phosphoreum was fully protected by P. phosphoreum mRNA from the S1 nuclease mapping assay suggesting that a transcriptional terminator was not present in the region. In addition, the levels of riboflavin synthase activity in P. phosphoreum was increased during the development of bacterial bioluminescence in the same fashion as the luciferase and fatty acid reductase activities. Insertion of the Photobacterium leiognathi DNA extending from luxB to ribII, between a strong lux promoter and a reporter gene (chloramphenicol acetyltransferase, CAT) and transferred by conjugation into P. leiognathi, did not affect expression of reporter gene. Moreover the CAT gene was not expressed in an analogous construct missing the lux promoter indicating that a promoter was not present in this region. Based on the data here, it can be concluded that the lux genes and rib genes in Photobacterium species are under common regulation.

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Effects of Adenophora triphylla Ethylacetate Extract on mRNA Levels of Antioxidant Enzymes in Human HepG2 Cells (인간 HepG2 Cell에서 항산화 효소의 mRNA 발현에 대한 잔대 에틸아세테이트 추출물 효과)

  • Choi, Hyun-Jin;Kim, Soo-Hyun;Oh, Hyun-Taek;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.10
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    • pp.1238-1243
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    • 2008
  • The root of Adenophora triphylla is widely used as traditional herbal medicine in Korea. We studied its effects on sodium nitroprusside (SNP) cytotoxicity and antioxidant genes expression in HepG2 cells. To study whether Adenophora triphylla ethylacetate extract (ATea) inhibited NO-induced cell death, HepG2 cells were preincubated for 24 hr with 50 and 100 $\mu$g/mL ATea followed by 24-hr exposure to 0.5 mM SNP (exogenous NO donor). No-induced cytotoxicity was inhibited by pretreatment of ATea, as assessed by mitochondrial dehydrogenase activity (MTT assay). We further investigated the effects of ATea on mRNA levels of various enzymes of the antioxidant system such as Cu, Zn superoxide dismutase (SOD 1), Mn SOD (SOD 2), glutathione peroxidase (GPx), catalase and several enzymes of the glutathione metabolism [glutathione reductase (GR), $\gamma$-glutamyl-cystein synthetase (GCS), glutathione-S-transferase (GST), $\gamma$-glutamyltranspeptidase ($\gamma$-GT), glucose-6-phosphate dehydrogenase (G6PD)] by RT-PCR. CAT, GCS, GR and G6PD mRNA levels were increased after treatment with ATea. The SOD 1, SOD 2, GPx, GST and $\gamma$-GT mRNA levels were not affected in ATea-treated HepG2 cells. We concluded that ATea have an indirect antioxidant effects, perhaps via induction of CAT, GCS, GR and G6PD.