• Title/Summary/Keyword: basal expression level

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Effects of Chromium Picolinate Supplementation on Growth Hormone Secretion and Pituitary mRNA Expression in Finishing Pigs

  • Wang, M.Q.;He, Y.D.;Xu, Z.R.;Li, W.F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.7
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    • pp.1033-1037
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    • 2008
  • The purpose of the present study was to investigate the effects of chromium picolinate (CrPic) on growth hormone (GH) secretion and pituitary GH mRNA expression in finishing pigs. Forty eight crossbred pigs with an initial body weight of 65.57 kg (SD = 1.05) were blocked by body weight and randomly assigned to two treatments with three replicates. Each group was fed the diet supplemented with or without $200{\mu}g/kg$ chromium from CrPic for 40 days. The results showed that average daily gain of pigs was increased by 9.84% (p<0.05), and longissimus muscle area was increased by 17.29% (p<0.05) with the supplementation of CrPic. The results of GH dynamic secretion showed that supplemental CrPic increased the mean level and peak value of GH by 36.58% (p<0.05) and 26.60% (p<0.05), respectively, while there was no significant effect on basal value, peak amplitude and peak duration. Pituitary mRNA expression of GH was not significantly influenced by supplemental CrPic. These results indicated that CrPic increased pigs GH secretion without change of pituitary GH mRNA expression.

Effects of Cheunggihwadamhwan Extract on Lowering Lipid, Antioxidation and Production of Proinflammatory Cytokines in Rats Fed on High Fat Diet (청기화담환(淸氣化痰丸) 추출물이 비만쥐의 지질강하, 항산화효과 및 전염증성 Cytokines 생산에 미치는 영향)

  • Lee, Han;Park, Won-Hyung;Cha, Yun-Yeop
    • Journal of Korean Medicine Rehabilitation
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    • v.24 no.2
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    • pp.1-13
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    • 2014
  • Objectives The purpose of this study was to investigate the effects of Cheunggihwadamhwan (CGHDH) extract on lowering lipid, antioxidation and production of proinflammatory cytokines in rats fed on high fat diet. Methods 40 Male Sprague-Dawley rats were fed on high fat diet for 8 weeks and 32 rats (above 400 g) were randomly divided into 4 groups (8 mice in each group) : control group, 100 mg/Kg CGHDH group, 200 mg/Kg CGHDH group, 300 mg/Kg CGHDH group. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And We fed each experimental group of rats basal diet and administered an extract of Cheunggihwadamhwan extracts (100 mg/kg, 200 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid in plasma and liver, concentration of proinflammatory cytokines, antioxidative activity and gene expression. The gene expression level was investigated by the way of reverse transcription-polymerase chain reaction (RT-PCR). Results 1. Concentration of plasma FFA, plasma TG, plasma total cholesterol and plasma LDL-cholesterol showed a significant decrement in Cheunggihwadamhwan groups. However, concentration of plasma HDL-cholesterol showed a significant increment in 200, 300 mg/kg Cheunggihwadamhwan groups. 2. Concentration of liver total cholesterol and liver TG showed a significant decrement in Cheunggihwadamhwan groups. 3. Concentration of plasma TBARS showed a significant decrement in all Cheunggihwadamhwan groups. Concentration of liver TBARS showed a significant decrement in 200, 300 mg/kg Cheunggihwadamhwan groups. Concentration of liver GSH-Px, SOD and CAT showed a tendency to decrease in all Cheunggihwadamhwan groups. 4. Concentration of plasma IL-$1{\beta}$, plasma IL-6, TNF-$\alpha$ and NO, showed a tendency to decrease in all Cheunggihwadamhwan groups. Concentration of plasma IL-10 showed a tendency to increase in all Cheunggihwadamhwan groups. 5. In the analysis of reverse transcription-polymerase chain reaction (RT-PCR), the gene expression of Apo-B and Apo-E in the Cheunggihwadamhwan groups showed a low expression than that of control group. The ratio of Apo-B expression per $\beta$-actin expression in the showed a significant decrement in all Cheunggihwadamhwan groups. The ratio of Apo-E expression per $\beta$-actin expression in the showed a significant decrement in 300 mg/kg Cheunggihwadamhwan groups. Conclusions According to this study, the extract of Cheunggihwadamhwan showed a positive effect of lowering lipid, antioxidation and a control of producing proinflammatory cytokines.

cDNA Cloning and Expression of a Cytochrome P450 1A (CYP1A) from the Pale Chub, Zacco platypus

  • Jeon, Hyoung-Joo;Park, Young-Chul;Lee, Wan-Ok;Lee, Jong-Ha;Kim, Jin-Hyoung
    • Korean Journal of Ecology and Environment
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    • v.44 no.4
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    • pp.364-372
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    • 2011
  • The pale chub (Zacco platypus) is generally found in Asian countries, such as Korea, Japan, and China. Nevertheless, very little information exists about the genes involved in the metabolism of xenobiotics in this species. This species is useful in monitoring the environmental impact on various pollutants in freshwater as a sentinel fish species. We cloned the full-length cDNA sequence of xenobiotic metabolizing cytochrome P450 1A (CYP1A) gene from Z. platypus and characterized it. Tissue distribution and timedependent induction of CYP1A were studied by real-time RT-PCR. Induction pattern of CYP1A was studied by exposing the fish to an arylhydrocarbon receptor agonist, ${\beta}$-naphthoflavone (BNF). The liver showed the highest level of expression in basal state as well as BNF- treated fish. However, appreciable levels of expression were also recorded in Gill and kidney and the least level of expression was observed in the eye. The results of the time-course study revealed an induction in the liver, brain, and gills after 6 h and 12 h in most of the tissues. This study provides an insight into the xenobiotics metabolizing system of Z. platypus and offers baseline information for further research related to biomarker, stress, and adaptive response of this ecologically important fish species in the freshwater environment.

Nam and Kim #1 Tissue-specific expression of pFV4CAT in transgenic mud loach (Misgurnus mizolepis) germ line (미꾸라지(Misgurnus mizolepis)에서 pFV4CAT 의 조직 특이적 발현)

  • Nam, Yoon-Kwon;Kim, Dong-Soo
    • Korean Journal of Ichthyology
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    • v.9 no.1
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    • pp.91-98
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    • 1997
  • The transgene, pFV4CAT, containing CAT reporter gene regulated by carp $\beta$-actin promoter, was expressed in independent transgenic mud loach germ lines, determined by reverse transcriptase-PCR (RT-PCR) and enzyme-linked immunosorbant assay (ELISA). Expression of the transmitted transgene was found to be tissue-specific in F1 and F2 generations. Tissue specificity of the expression was dependent on each transgenic line with reproducible patterns. Liver and spleen did express the transgene more frequently than other tissues tested, and muscle and heart revealed the higher amount of CAT than other tissues, while testes showed the lowest expression level. The highest level of CAT expression in muscle from a transgenic F1 line was corresponding to 68-fold compared to the basal levels of controls.

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Effect of Chitosan on Nitric Oxide Content and Inducible Nitric Oxide Synthase Activity in Serum and Expression of Inducible Nitric Oxide Synthase mRNA in Small Intestine of Broiler Chickens

  • Li, H.Y.;Yan, S.M.;Shi, B.L.;Guo, X.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.7
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    • pp.1048-1053
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    • 2009
  • The present study was conducted to determine the effects of chitosan on nitric oxide (NO) content and inducible nitric oxide synthase (iNOS) activity in serum, and relative expression of iNOS mRNA in the duodenum, jejunum, and ileum of broiler chickens. A total of 240 one-day-old Arbor Acre mixed-sex broiler chickens were randomly allotted to six dietary treatments with five replicates in each treatment and eight chickens in each replicate. The broiler chickens in the six treatments were fed the basal diet supplemented with 0 (control), 0.05, 0.2, 0.5, 1.0 or 2.0 g/kg chitosan. The trial lasted for 42 days. The results showed that dietary chitosan enhanced NO content and iNOS activity in serum as well as iNOS mRNA expression in the duodenum and ileum of broiler chickens in a quadratic dose-dependent manner (p<0.05), and improved jejunum iNOS mRNA expression in a quadratic dose-dependent manner (p<0.10) with increasing addition of chitosan. Chicks fed a diet containing 0.5-1.0 g/kg chitosan had higher NO content and iNOS activity in serum as well as small-intestinal iNOS mRNA expression compared with birds given the control diet, but positive effects of chitosan tended to be suppressed when addition of chitosan in the diet was increased to 2.0 g/kg. These results implied that there was a threshold level of chitosan inclusion beyond which progressive reductions in serum NO content and small intestinal iNOS expression occured, and the regulation of chitosan on immune functions in chickens is probably associated with activated expression of iNOS and NO secretion.

High fat diet confers vascular hyper-contractility against angiotensin II through upregulation of MLCK and CPI-17

  • Kim, Jee In
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.1
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    • pp.99-106
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    • 2017
  • Obesity is a critical risk factor for the hypertension. Although angiotensin II (Ang II) in obese individuals is known to be upregulated in obesity-induced hypertension, direct evidence that explains the underlying mechanism for increased vascular tone and consequent increase in blood pressure (BP) is largely unknown. The purpose of this study is to investigate the novel mechanism underlying Ang II-induced hyper-contractility and hypertension in obese rats. Eight-week old male Sprague-Dawley rats were fed with 60% fat diet or normal diet for 4 months. Body weight, plasma lipid profile, plasma Ang II level, BP, Ang II-induced vascular contraction, and expression of regulatory proteins modulating vascular contraction with/without Ang II stimulation were measured. As a result, high fat diet (HFD) accelerated age-dependent body weight gaining along with increased plasma Ang II concentration. It also increased BP and Ang II-induced aortic contraction. Basal expression of p-CPI-17 and myosin light chain (MLC) kinase was increased by HFD along with increased phosphorylation of MLC. Ang II-induced phosphorylation of CPI-17 and MLC were also higher in HFD group than control group. In conclusion HFD-induced hypertension is through at least in part by increased vascular contractility via increased expression and activation of contractile proteins and subsequent MLC phosphorylation induced by increased Ang II.

Keumsa Linteusan Suppresses Invasion of Cancer Cells through the Inhibition of Cellular Adhesion and MMP-9 Expression

  • Kim, Won-Jung;Hong, Sung-Chang;Do, Eun-Ju;Suk, Kyung-Ho;Yun, lk-Jin;Lee, Won-Ha
    • Animal cells and systems
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    • v.13 no.2
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    • pp.113-118
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    • 2009
  • Extracts derived from various medical mushrooms have been reported to have antitumor and immuno-modulatory properties. In order to investigate the antitumor activity of keumsa Linteusan, the water extract of Phellinus Iimteus, HT1080 cells, a human fibrosarcoma cell line, were treated with it and changes in cellular migration potential was tested in vitro. At a concentration range below 1,000 $\mu$g/mL, Linteusan blocked, in a dose dependent manner, the migration of cells through Matrigel as well as Boyden chamber without affecting the viability of the cells. Prolonged treatment of HT1080 cells with Linteusan suppressed TNFa induced production of matrix metalloproteinase (MMP)-9 as well as basal level expression of MMP-2. Linteusan also affected the adhesion of the cells to fibronectin-coated surfaces. The effect of Linteusan on cell signaling pathways was also tested. Linteusan specifically affected TNF-$\alpha$ induced phosphorylation of AKT in a dose-dependent manner, while phosphorylation levels of ERK remained unaffected. These data indicate that Linteusan blocks the migration of HT1080 cells by affecting various processes associated with cell migration such as the expression of matrix degradingenzymes,cell adhesion, and AKT-medicated cellular signaling pathways.

Iron Chelator-Inducible Expression System for Escherichia coli

  • Lim, Jae-Myung;Hong, Mi-Ju;Kim, Seong-Hun;Oh, Doo-Byoung;Kang, Hyun-Ah;Kwon, Oh-Suk
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1357-1363
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    • 2008
  • The $P_{entC}$ promoter of the entCERA operon encoding enzymes for enterobactin biosynthesis in Escherichia coli is tightly regulated by the availability of iron in the culture medium. In iron-rich conditions, the $P_{entC}$ promoter activity is strongly repressed by the global transcription regulator Fur (ferric uptake regulator), which complexes with ferrous ions and binds to the Fur box 19-bp inverted repeat. In this study, we have constructed the expression vector pOS2 containing the $P_{entC}$ promoter and characterized its repression, induction, and modulation by quantifying the expression of the lacZ reporter gene encoding $\beta$-galactosidase. $\beta$-Galactosidase activities of E. coli transformants harboring pOS2-lacZ were highly induced in the presence of divalent metal ion chelators such as 2,2'-dipyridyl and EDTA, and were strongly repressed in the presence of excess iron. It was also shown that the basal level $\beta$-galactosidase expression by the $P_{entC}$ promoter was drastically decreased by incorporating the fur gene into the expression vector. Since the newly developed iron chelator-inducible expression system is efficient and cost-effective, it has wide applications in recombinant protein production.

Exosome-derived microRNA-29c Induces Apoptosis of BIU-87 Cells by Down Regulating BCL-2 and MCL-1

  • Xu, Xiang-Dong;Wu, Xiao-Hou;Fan, Yan-Ru;Tan, Bing;Quan, Zhen;Luo, Chun-Li
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.8
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    • pp.3471-3476
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    • 2014
  • Background: Aberrant expression of the microRNA-29 family is associated with tumorigenesis and cancer progression. As transport carriers, tumor-derived exosomes are released into the extracellular space and regulate multiple functions of target cells. Thus, we assessed the possibility that exosomes could transport microRNA-29c as a carrier and correlations between microRNA-29c and apoptosis of bladder cancer cells. Materials and Methods: A total of 28 cancer and adjacent tissues were examined by immunohistochemistry to detect BCL-2 and MCL-1 expression. Disease was Ta-T1 in 12 patients, T2-T4 in 16, grade 1 in 8, 2 in 8 and 3 in 12. The expression of microRNA-29c in cancer tissues was detected by quantitative reverse transcriptase PCR (QRT-PCR). An adenovirus containing microRNA-29c was used to infect the BIU-87 human bladder cancer cell line. MicroRNA-29c in exosomes was measured by QRT-PCR. After BIU-87 cells were induced by exosomes-derived microRNA-29c, QRT-PCR was used to detect the level of microRNA-29c. Apoptosis was examined by flow cytometry and BCL-2 and MCL-1 mRNA expressions were assessed by reverse transcription-polymerase chain reaction. Western blotting was used to determine the protein expression of BCL-2 and MCL-1. Results: The expressions of BCL-2 and MCL-1 protein were remarkably increased in bladder carcinoma (p<0.05), but was found mainly in the basal and suprabasal layers in adjacent tissues. The expression of microRNA-29c in cancer tissues was negatively correlated with the BCL-2 and MCL-1. The expression level of microRNA-29c in exosomes and BIU-87 cells from the experiment group was higher than that in control groups (p<0.05). Exosome-derived microRNA-29c induced apoptosis (p<0.01). Although only BCL-2 was reduced at the mRNA level, both BCL-2 and MCL-1 were reduced at the protein level. Conclusions: Human bladder cancer cells infected by microRNA-29c adenovirus can transport microRNA-29c via exosomes. Moreover, exosome-derived microRNA29c induces apoptosis in bladder cancer cells by down-regulating BCL-2 and MCL-1.

Regulation of Immediate Early Gene Expression by Glutamate Receptor Activation in C6 Rat Glioma Cells

  • Lee, Jin-Koo;Kim, Yung-Hi;Choi, Seong-Soo;Suh, Hong-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.1
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    • pp.19-25
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    • 2001
  • We have studied the effects of excitatory amino acids on the expression of the c-fos and c-jun mRNA in rat C6 glioma cells. The glutamate, $N-methyl-_D-aspartate$ (NMDA), and kainic acid (KA) increased c-fos mRNA level in a concentration-dependent manner. However, they did not affect c-jun mRNA level. In addition, forskolin and phorbol 12-myristate 13-acetate (PMA) increased c-fos mRNA level. Furthermore, PMA increased c-jun mRNA level whereas forskolin downregulated c-jun mRNA level. The glutamate, NMDA and KA, at a concentration of 0.25 mM, did not affect the basal c-fos and c-jun mRNA levels, and also did not affect forskolin- and PMA-induced responses. Furthermore, both forskolin and PMA itself increased the phosphorylation of ERK (extracellular signal regulated kinase) and CREB (cyclicAMP responsible element binding protein) proteins. The KA, NMDA, and glutamate did not affect forskolin- induced increase of ERK and CREB phosphorylation. The KA decreased PMA-induced increase of phosphorylation of ERK and CREB proteins, whereas glutamate and NMDA did not affect the phosphorylation of ERK and CREB proteins induced by PMA. These findings suggest that, in C6 glioma cells, c-fos mRNA induction induced by EAAs is not mediated by phosphorylation of ERK and CREB proteins.

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