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Effect of Dietary Supplementation of Acanthopanax senticosus and Eucommiaceae on the Expression of Lipogenic, Myogenic and Antioxidant Enzyme Genes in Broiler Chickens (육계에서 가시오갈피와 두충의 첨가 급여가 항산화 효소, 지방 및 근육 관련 유전자 발현에 미치는 영향)

  • Kang, H.K.;Beloor, J.;Sohn, S.H.;Jang, I.S.;Moon, Y.S.
    • Korean Journal of Poultry Science
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    • v.36 no.1
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    • pp.39-45
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    • 2009
  • This study was carried out to investigate the effect of dietary supplementation of Acanthopanax (A) senticosus and Eucommiaceae on the expression of lipogenic, myogenic and oxidative stress genes in broiler chickens. Birds were subjected (assigned) to one of the following 5 dietary treatments: control (CON), A. senticosus 0.5% (T1), 1.0% (T2), Eucommiaceae 0.5% (T3) and 1% (T4). Each treatment was replicated 8 times with 4 birds per replication, housed in 4 birds per cage. Birds were arranged according to randomized block design. Feeding trial was conducted from day 4 to 35th day of age. Liver and muscle tissues were collected for analysis. Broilers subjected to 1% A. senticosus had higher feed conversion ratio than the other treated birds whereas no significant differences were found in body weight, weight gain and feed intake. The gene expression levels of fatty acid synthase were not different among the treatments while the transcription factor $PPAR{\gamma}$ was highly expressed in Eucommiaceae but not in control and A. senticosus. The gene expression levels of myogenin were high in both A. senticosus and Eucommiaceae compared to control group. MyoD also showed high expression in treated groups furthermore, Eucommiaceae stimulated the expression of MyoD more than that of A. senticosus. The antioxidant gene expressions (SOD, CAT, SOD, GPX) generally were not much different among the treatments, however, SOD and GPX were stimulated in broilers consumed 1% Eucommiaceae diet. The result of this experiment showed that dietary supplementation of A. senticosus and Eucommiaceae in broiler may improve the antioxidant defence system through SOD and GPX without affect of growth performance in broilers.

Effect of the Contents Ratio of Panaxadiol Ginsenosides Extracted from Various Compartment of Ginseng on the Transcription of Cu/Zn Superoxide Dismutase Gene (홍삼의 각 부위에서 추출된 Panaxadiol분획의 함량비에 따른 유해산소제거효소(Cu/Zn Superoxide Dismutase) 유도효과)

  • Chang Mun Seog;Choi Kang Ju;Rho Hyune Mo
    • Journal of Ginseng Research
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    • v.23 no.1 s.53
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    • pp.44-49
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    • 1999
  • Cu/Zn superoxide dismutase (SOD1) is a protective enzyme responsible for the dismutat ion of superoxide radicals within the cell by converting superoxide radicals to oxygen and hydrogen peroxide, which is in turn changed to oxygen and water by catalase. Previously, we reported that the panaxadiol (PD) and its ginsenoside $Rb_2$ induced the expression of SOD1 gene through AP2 binding site and its induction. Here, we examined the effect of subfractions of panaxadiol ginsenosides, which were extracted from different parts of ginseng root that possess various ratios of panaxadiol to panaxatriol, on the induction of SOD1 gene expression. To explore this possibility, the upstream regulatory region of SOD1 was linked to the chloramphenicol acetyl transferase (CAT) structural gene and introduced into human hepatoma HepG2 cells. We observed that the transcriptional activation of SOD1 was proportional to the contents ratio of panaxadiol ginsensides. Consistent with this results, the total extract portion prepared from the finely-hairy root, which contains the higher ratio of panaxadiol to panaxatriol about 2.6, increased the SODl transcription about 3 fold. This results suggest that the panaxadiol fraction could induce the SOD1 and total extract of the ginseng finely-hairy root would be a useful material as a functional food for the SOD1 inducer.

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Induction of apoptosis by etoposide treatment in colon cancer cell line SNU C2A (대장암 세포주 SNU C2A에서 etoposide 처리에 의한 apoptosis 유도)

  • Jung, Ji-Yeon;Na, Yun-sook;Jung, Ho-Chul;Oh, Sang-Jin
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.221-229
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    • 2001
  • Background: Inactivation of tumor suppressor genes is believed to be important in the development of many human malignancies. Recently, several lines of evidence have indicated that the wild type p53 gene located at 17p13.3, may function as a tumor suppressor gene and that a mutant p53 gene could promote transformation by inactivating normal p53 function in a dominant negative fashion. These broad spectrum of p53 mutation in human cancers provide that mutant p53 and their protein may be potential targets of tumor diagnostic and therapeutic interventions. Method: Colony formation was performed to investigate growth suppressional ability. p53 expression pattern was examined by western blot and p53-mediated transactivation ability was assessed by CAT activity. SNU C2A cells were observed in apoptotic aspects induced by etoposide and $H_2O_2$ treatment, detecting sensitivity on agent, DNA fragmentation through agarose gel, chromatin condensation by fluorescence microscope, and cell cycle distribution by FACS. Result: 1) p53 mutant his179arg ($histidine{\rightarrow}arginine$) detected in SNU C2A cells lost transcriptional activity and growth suppression ability, showing dominant negative effect on its wild type p53. 2) Etoposide-treated SNU C2A cells induced apoptosis, exhibiting dramatic reduction of cell growth, DNA fragmentation, nuclear condensation formation of apoptotic body and increment of sub-G1 cell fraction. 3) Etoposide and $H_2O_2$-treated SNU C2A cells have no high increase of p53 expression and overexpressed p53 protein changed localization, from cytoplasm to nucleus. Also, p53-mediated transcriptional activity was increased by agents-treatment. Conclusion: SNU C2A cells coexpress wild-type and mutant p53 protein induced apoptosis in the condition on DNA damage, through localizational shift from cytoplasm to nucleus of p53 protein rather than the induction of p53 protein. SNU C2A cells derived mutant p53 his179arg abrogated both the growth supression ability and transactivational activity, showing inhibition effect on transcriptional activity of wild type p53, but did not repress the activity of wild type p53 in SNU C2A cells owing to dominant activity of wild type. These cell condition may provide new gene therapeutic implications leading effective antiproliferation of cell when mutant and wild-type p53 protein were co-expressed in cell.

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Identification of a Novel Cassette Array in Integron-bearing Helicobacter Pylori Strains Isolated from Iranian Patients

  • Goudarzi, Mehdi;Seyedjavadi, Sima Sadat;Fazeli, Maryam;Roshani, Maryam;Azad, Mehdi;Heidary, Mohsen;Navidinia, Masoumeh;Goudarzi, Hossein
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3309-3315
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    • 2016
  • Helicobacter pylori as the second most common cause of gastric cancer in the world infects approximately half of the developed countries population and 80% of the population living in developing countries. Integrons as genetic reservoirs play major roles in dissemination of antimicrobial resistance genes. To the best of our knowledge, this is the first study to report carriage of class 1 and 2 integrons and associated gene cassettes in H. pylori isolates from Iran. This cross-sectional study was conducted in Tehran among 110 patients with H. pylori infection. Antimicrobial susceptibility testing (AST) for H. pylori strains were assessed by the micro broth dilution method. Class 1 and 2 integrons were detected using PCR. In order to determine gene cassettes, amplified fragments were subjected to DNA sequencing of both amplicon strands. The prevalence of resistance to clarithromycin, metronidazole, clarithromycin, tetracycline, amoxicillin, rifampin, and levofloxacin were 68.2% (n=75), 25.5% (n=28), 24.5% (n=27), 19.1% (n=21), 18.2% (n=20) and 16.4% (n=18), respectively. Frequency of multidrug resistance among H. pylori isolates was 12.7%. Class 2 integron was detected in 50 (45.5%) and class 1 integron in 10 (9.1%) H. pylori isolates. The most predominant gene cassette arrays in class 2 integron-bearing H. pylori were included sat-era-aadA1, dfrA1-sat2-aadA1, blaoxa2 and, aadB whereas common gene cassette arrays in class 1 integron were aadB-aadA1-cmlA6, aacA4, blaoxa2, and catB3. The high frequency of class 2 integron and multidrug resistance in the present study should be considered as a warning for clinicians that continuous surveillance is necessary to prevent the further spread of resistant isolates.

sanN Encoding a Dehydrogenase is Essential for Nikkomycin Biosynthesis in Streptomyces ansochromogenes

  • Ling, Hong-Bo;Wang, Guo-Jun;Li, Jin-E;Tan, Hua-Rong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.397-403
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    • 2008
  • Nikkomycins are a group of peptidyl nucleoside antibiotics with potent fungicidal, insecticidal, and acaricidal activities. sanN was cloned from the partial genomic library of Streptomyces ansochromogenes 7100. Gene disruption and complementation analysis demonstrated that sanN is essential for nikkomycin biosynthesis in S. ansochromogenes. Primer extension assay indicated that sanN is transcribed from two promoters (sanN-P1 and sanN-P2), and sanN-P2 plays a more important role in nikkomycin biosynthesis. Purified recombinant SanN acts as a dehydrogenase to convert benzoate-CoA to benzaldehyde in a random-order mechanism in vitro, with respective $K_{cat}/K_m$$ values of $3.8mM^{-1}s^{-1}\;and\;12.0mM^{-1}s^{-1}$ toward benzoate-CoA and NADH, suggesting that SanN catalyzes the formation of picolinaldehyde during biosynthesis of nikkomycin X and Z components in the wild-type stain. These data would facilitate us to understand the biosynthetic pathway of nikkomycins and to consider the combinatorial synthesis of novel antibiotic derivatives.

Experimental Study of Chungganhaeju-tang (Qingganjiejiu-tang) on Oxidative Stress (청간해주탕(淸肝解酒湯)의 항산화 작용에 관한 실험적 연구)

  • Lee, Ji-Eun;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.32 no.2
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    • pp.188-202
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    • 2011
  • Objectives : Oxidative stress seems to play a major role in mechanisms by which ethanol causes liver injury. Previous studies have shown that treatment with Chungganhaeju-tang (Qingganjiejiu-tang, CGHJT) has protective effects on alcoholic liver disease. The aim of this study was to investigate the effects of Chungganhaeju-tang on oxidative stress. Materials and Methods : In vitro, we evaluated the inhibitory activities of CGHJT on DPPH (1,1-diphenyl-2-picryl-hydrazyl), xanthine oxidase, trypsin, and hyaluronidase, and measured cell viability, and proliferation. In the cell culture model, we measured the activities of superoxide dismutase (SOD), and catalase (CAT) after CGHJT treatment in C34 and E47 cell lines, HepG2 cells transfected with/without the cytochrome P450 2E1 (CYP2E1) gene. In vivo, we measured malondialdehyde levels in the liver tissue and alcohol concentration in the blood. Results : CGHJT showed significant free radical scavenging activity against DPPH and xanthine oxidase in the in vitro study, and increased cell viability, proliferation, and activities of superoxide dismutase, catalase in C34 and in E47 cell lines. CGHJT reduced malondialdehyde levels and blood alcohol concentration in vivo, as well. Conclusions : This study suggests that CGHJT has antioxidant effects on oxidative stress by reducing lipid peroxidation and inhibiting the ethanol induced suppression of antioxidant enzyme activities.

Purification and Comparison of Properties of the C-Terminus Truncated Agarase of Pseudomonas sp. W7

  • Yoon, Soo-Cheol;Lee, Jong-Hee;Ahn, Sun-Hee;Lee, Eun-Mi;Park, Eun-Mi;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.767-772
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    • 2003
  • Three plasmids derived from the ${\beta}-agarase$ gene (PjaA) of Pseudomonas sp. W7 were expressed in Escherichia coli AD494(DE3) pLysS with lactose as an inducer. These products corresponded to the complete (PjaA) and the two C-terminal truncated (PjaAI and PjaAII) forms of ${\beta}-agarase$. The PjaAI and the PjaAII were originated from exonuclease L treatment from PjaA by deleting 127 and 182 amino acid residues-encoded nucleic acids at 3' region, respectively. The molecular weights of the purified proteins were 71 kDa, 58 kDa, and 50 kDa on SDS-PAGE, respectively. The $K_m$ value of PjaAI was lower than that of the PjaA, and the catalytic efficiency ($k_{cat}/K_m$) of PjaAI was increased to 5 times. The enzyme of PjaAI retained more than 90% activity at $50^{\circ}C$. In contrast to the PjaAI, the remaining activity of the PjaA was only 20% at the same temperature.

Effect of reactive oxygen species on floral senescence in Hibiscus syriacus L. (활성산소종이 무궁화 꽃의 노화에 미치는 영향)

  • Seo, Sang-Kyu;Kim, Sun-Hyung;Lee, Gung-Pyo;Kang, Seung-Won
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.179-183
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    • 2009
  • To understand the effect of reactive oxygen species (ROS) on floral senescence in Hibiscus syriacus L., we have investigated change in relative water potential, malondialdehyde (MDA) content, H_2O_2 content and the activity of antioxidative enzymes in the petals during flower opening and senescence. Hibiscus flowers were achieved full bloom at early morning and started to in-rolling and showed petal in-rolling over than 50% at 24 h and 36 h after full bloom, respectively. The flower was a decrease in fresh weight by 30% and showed water loss with floral senescence. MDA content and activity of antioxidative enzymes such as APX, GR and CAT were showed no significant change until 36 h after full bloom. In the flower 48 h after full bloom that showed complete petal in-rolling and wilting, however, activity of antioxidative enzymes and H_2O_2 content was greatly increased as compared with 0 h after full bloom. These results suggest that reactive oxygen species are related to accelerating the later senescence more than inducing the early senescence during Hibiscus flower senescence.

Cloning, Expression, and Purification of Exoinulinase from Bacillus sp. snu-7

  • Kim, Kyoung-Yun;Koo, Bong-Seong;Jo, Do-Hyun;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.344-349
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    • 2004
  • A gene encoding inulin-degrading enzyme of Bacillus sp. snu-7 with ORF of 1536 nucleotides was cloned. And it was overexpressed as His-tagged protein in E. coli BL21(DE3) pLysS using pRSET B vector containing mature enzyme sequence. Maximum enzyme production was achieved by IPTG (0.1 mM) induction at $OD_{600}$ 1.2 and $30^{\circ}C$ followed by 6 h incubation. The expressed protein purified through immobilized metal affinity chromatography showed molecular mass of 60 kDa on SDS-PAGE. Results of thin-layer chromatography using inulin as a substrate showed the enzyme to be an exotype inulinase capable of producing only monomeric fructose as a product. $K_m$ and $k_{cat}$, for the hydrolyses of inulin and sucrose were $2.28\pm0.08$ mM and 358.05$\pm$20.38 $min^{-l}$, and 22.02$\pm$0.41 mM and 4619.11$\pm$215.12 $$min^{-1}, respectively. Optimal activity of the exoinulinase occurred at pH 7.0 and $50^{\circ}C$.

Genotype of Toxoplasma gondii from Blood of Stray Cats in Gyeonggi-do, Korea

  • Kim, Hye-Youn;Kim, Yun-Ah;Lee, Ho-Sa;Rhie, Ho-Gun;Cho, Shin-Hyeong;Yu, Jae-Ran;Lee, Sang-Eun
    • Parasites, Hosts and Diseases
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    • v.47 no.4
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    • pp.413-415
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    • 2009
  • Genotyping of Toxoplasma gondii has been performed in 23 PCR positive blood samples from stray cats in Korea. We used 2 separate PCR-restriction fragment length polymorphism (RFLP) patterns of SAG2 gene, amplifying the 5' and 3' ends of the locus. The results revealed that all samples belonged to the type I clonal lineage. Although T. gondii organisms were not isolated from the samples, the results of the present study represent that stray cats with T. gondii infection should be seriously concerned in our environment. Adequate and continuous control programs of stray cats are needed to reduce the risk of transmission of T. gondii as a zoonotic infection threatening the public health.