• Title/Summary/Keyword: Marker compound

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Tissue Culture Studies of Anthranilate Synthase the Tryptophan Biosynthetic Control Enzyme

  • Widholm, Jack.M.
    • Journal of Plant Biotechnology
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    • v.2 no.2
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    • pp.55-60
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    • 2000
  • Experiments initiated 30 years ago to obtain selectable markers have led to a series of studies of Trp biosynthesis and anthranilate synthase (AS) the control enzyme using largely plant tissue cultures since they have experimental properties that can be readily exploited. Enzymological and compound feeding studies provided evidence that AS is the control point in the Trp biosynthesis branch and that altering the AS feedback control by the selection of mutants resistant to the Trp analog 5-methyl-tryptophan (5MT) can lead to the overproduction of this important amino acid. Plants regenerated from these Trp overproducing lines of most species also had high free Trp levels but Nicotiana tabaum (tobacco) plants expressed the feedback altered AS only in cultured cells and not in the regenerated plants. further tests by transient and stable expression of the cloned promoter for the naturally occurring tobacco feedback-insensitive AS, denoted ASA2, confirmed the tissue culture specific nature of the expression control. The 5MT caused by the expression of a feedback-insensitive AS from tobacco has been used to select protoplast fusion hybrids with several species since the resistance is expressed dominantly. Recently the ASA2 gene has been used successfully as a selectable marker to select transformed Astragalus sinicus and Glycine max hairy roots induced by Agrobactetium rhizogenes. These results show that the ASA2y-subunit can interact with the y-subunit of another species to form active feedback-insensitive enzyme that may be useful for selecting transformed cells. Plastid DNA transformation of tobacco has also effectively expressed ASA2 in the compartment in which Trp biosynthesis is localized in the cell.

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Prognostic Significance of Altered Blood and Tissue Glutathione Levels in Head and Neck Squamous Cell Carcinoma Cases

  • Khan, Sami Ullah;Mahjabeen, Ishrat;Malik, Faraz Arshad;Kayani, Mahmood Akhtar
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7603-7609
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    • 2014
  • Glutathione is a thiol compound that plays an important role in the antioxidant defense system of the cell and its deficiency leads to an increased susceptibility to oxidative stress and, thus, progression of many disease states including head and neck cancer. In the present study, alterations of glutathione levels were investigated in study cohort of 500 samples (cohort 1 containing 200 head and neck cancer blood samples along with 200 healthy controls and cohort II with 50 head and neck squamous cell carcinoma tissue samples along with 50 control tissues) by high performance liquid chromatography. The results indicated that mean blood glutathione levels were significantly reduced in head and neck cancer patients (p<0.001) compared to respective controls. In contrast, the levels of glutathione total (p<0.05) and glutathione reduced (p<0.05) were significantly elevated in head and neck squamous cell carcinoma tissues compared to the adjacent cancer-free control tissues. In addition to this, pearson correlation performed to correlate different tissue glutathione levels (GSH) with clinical/pathological parameters demonstrated a significant negative correlation between pT-stage and GSH level ($r=-0.263^{**}$; p<0.01), C-stage and GSH level ($r=-0.335^{**}$; p<0.01), grade and GSH ($r=-0.329^{**}$; p<0.01) and grade versus redox index ($r=-0.213^{**}$; p<0.01) in HNSCC tissues. Our study suggests that dysregulation of glutathione levels in head and neck cancer has the potential to predict metastasis, and may serve as a prognostic marker.

Ginsenoside Rg3 promotes inflammation resolution through M2 macrophage polarization

  • Kang, Saeromi;Park, Soo-Jin;Lee, Ae-Yeon;Huang, Jin;Chung, Hae-Young;Im, Dong-Soon
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.68-74
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    • 2018
  • Background: Ginsenosides have been reported to have many health benefits, including anti-inflammatory effects, and the resolution of inflammation is now considered to be an active process driven by M2-type macrophages. In order to determine whether ginsenosides modulate macrophage phenotypes to reduce inflammation, 11 ginsenosides were studied with respect to macrophage polarization and the resolution of inflammation. Methods: Mouse peritoneal macrophages were polarized into M1 or M2 phenotypes. Reverse transcription-polymerase chain reaction, Western blotting, and measurement of nitric oxide (NO) and prostaglandin $E_2$ levels were performed in vitro and in a zymosan-induced peritonitis C57BL/6 mouse model. Results: Ginsenoside $Rg_3$ was identified as a proresolving ginseng compound based on the induction of M2 macrophage polarization. Ginsenoside $Rg_3$ not only induced the expression of arginase-1 (a representative M2 marker gene), but also suppressed M1 marker genes, such as inducible NO synthase, and NO levels. The proresolving activity of ginsenoside $Rg_3$ was also observed in vivo in a zymosan-induced peritonitis model. Ginsenoside $Rg_3$ accelerated the resolution process when administered at peak inflammatory response into the peritoneal cavity. Conclusion: These results suggest that ginsenoside $Rg_3$ induces the M2 polarization of macrophages and accelerates the resolution of inflammation. This finding opens a new avenue in ginseng pharmacology.

Protective effect of ginsenoside Re on acute gastric mucosal lesion induced by compound 48/80

  • Lee, Sena;Kim, Myung-Gyou;Ko, Sung Kwon;Kim, Hye Kyung;Leem, Kang Hyun;Kim, Youn-Jung
    • Journal of Ginseng Research
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    • v.38 no.2
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    • pp.89-96
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    • 2014
  • The protective effect of ginsenoside Re, isolated from ginseng berry, against acute gastric mucosal lesions was examined in rats with a single intraperitoneal injection of compound 48/80 (C48/80). Ginsenoside Re (20 mg/kg or 100 mg/kg) was orally administered 0.5 h prior to C48/80 treatment. Ginsenoside Re dose-dependently prevented gastric mucosal lesion development 3 h after C48/80 treatment. Increases in the activities of myeloperoxidase (MPO; an index of neutrophil infiltration) and xanthine oxidase (XO) and the content of thiobarbituric acid reactive substances (TBARS; an index of lipid peroxidation) and decreases in the contents of hexosamine (a marker of gastric mucus) and adherent mucus, which occurred in gastric mucosal tissues after C48/80 treatment, were significantly attenuated by ginsenoside Re. The elevation of Bax expression and the decrease in Bcl2 expression after C48/80 treatment were also attenuated by ginsenoside Re. Ginsenoside Re significantly attenuated all these changes 3 h after C48/80 treatment. These results indicate that orally administered ginsenoside Re protects against C48/80-induced acute gastric mucosal lesions in rats, possibly through its stimulatory action on gastric mucus synthesis and secretion, its inhibitory action on neutrophil infiltration, and enhanced lipid peroxidation in the gastric mucosal tissue.

Anti-allergic Effect of Zizania latifolia Turcz Extracts (고장초의 항알레르기 효과)

  • Lee, Eun-Jung;Whang, Eun-Yeong;Whang, Key;Lee, In-Seon;Yang, Seun-Ah
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.717-721
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    • 2009
  • The anti-allergic effect of the methanol extract of Zizania latifolia Turcz was investigated. Methanol extract of Z. latifolia Trucz did not display any cytotoxicity activities in the IgE-sensitized RBL-2H3 cells. The extract was found to inhibit compound 48/80-induced degranulation and antigen-induced $\beta$-hexosaminidase release as well as PMA plus A23187-induced TNF-$\alpha$ production in RBL-2H3 mast cells in a dose-dependent manner. Release of $\beta$-hexosaminidase, a marker for the release of histamine in mast cells, was inhibited with an $IC_{50}$ value of $73.8\;{\mu}g/mL$. These results suggest that Z. latifolia extracts may be useful for the prevention of type I allergic reaction.

Studies on the Crude Drugs of Atractylodis Species (출류(朮類) 대조 생약에 관한 연구)

  • Kim, Ho-Hyun;Chon, In-Ju;Kang, In-Ho;Ham, In-Hye;Ze, Keam-Ryon;Whang, Wan-Kyunn;Cho, Hyoung-Kwon
    • Korean Journal of Pharmacognosy
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    • v.34 no.2 s.133
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    • pp.123-127
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    • 2003
  • Atrartylodis Rhizoma(蒼朮)'s origin plants are Atratylodes lancea and A. Chinensis in Chinese, Japanese and Korean pharmacopoeia. A. Japonica is only indigenous in Korea, it is actually used as Atractylodis Rhizoma in Korean market. A. lancea is used in Hunan province, China and A. Chinensis is used in Hubei province, China. It is impossible to distinguish with species differency as macro- and micro-morphology. We tried to distinguish with species differency by HPLC and GC-Mass spectra. Atractylone(mw. 216) which is a marker compound in Atractylodis Rhizoma Alba(白朮) was detected in A. japonica. Atractylodin (mw.182) was detected in A. lancea and two eudesmadien derivatives (mw. 204) were detected in A. chinensis. HPLC chromatogram showed the same patterns. As a result, we propose that A. japonica will be added as Atractylodis Rhizoma (蒼朮)'s origin plant in Korean Pharmacopoeia. Atractylodis Rhizoma Alba(白朮)'s origin plants are A. macrocephala in China, and A. Japonica and A. ovata in Korea and Japan. In GC-Mass analysis, all samples showed same patterns and the main compound was atractylone.

Anti-adipogenic Pregnane Steroid from a Hydractinia-associated Fungus, Cladosporium sphaerospermum SW67

  • Lee, Seoung Rak;Kang, Heesun;Yoo, Min Jeong;Yu, Jae Sik;Lee, Seulah;Yi, Sang Ah;Beemelmanns, Christine;Lee, Jaecheol;Kim, Ki Hyun
    • Natural Product Sciences
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    • v.26 no.3
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    • pp.230-235
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    • 2020
  • A pregnane steroid, 3α-hydroxy-pregn-7-ene-6,20-dione (1), was isolated from a Hydractinia-associated Cladosporium sphaerospermum SW67 by repetitive column chromatographic separation and high-performance liquid chromatography (HPLC) purification. The planar structure of 1 was elucidated from the analysis of the spectroscopic data (1D and 2D NMR spectra) and LC-MS data. The absolute configuration of 1 was determined by interpretation of ROESY spectrum of 1, together with the comparison of reported spectroscopic values in previous studies. To the best of our knowledge, this is the first report of the identification of the pregnane scaffold from C. sphaerospermum, a natural source. Compound 1 was evaluated for its effects on lipid metabolism and adipogenesis during adipocyte maturation and showed that compound 1 substantially inhibited lipid accumulation compared to the control. Consistently, the expression of the adipocyte marker gene (Adipsin) was reduced upon incubation with 1. Further, we evaluated the effects of 1 on lipid metabolism by measuring the transcription of lipolytic and lipogenic genes. The expression of the lipolytic gene ATGL was significantly elevated upon exposure to 1 during adipogenesis, whereas the expression of lipogenic genes FASN and SREBP1 was significantly reduced upon treatment with 1. Thus, our findings provide experimental evidence that the steroid derived from Hydractinia-associated C. sphaerospermum SW67 is a potential therapeutic agent for obesity.

Induction of cytoprotective autophagy by morusin via AMP-activated protein kinase activation in human non-small cell lung cancer cells

  • Park, Hyun-Ji;Park, Shin-Hyung
    • Nutrition Research and Practice
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    • v.14 no.5
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    • pp.478-489
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    • 2020
  • BACKGROUND/OBJECTIVES: Morusin, a marker component of Morus alba L., possesses anti-cancer activity. The objective of this study was to determine autophagy-inducing effect of morusin in non-small cell lung cancer (NSCLC) cells and investigate the underlying mechanism. SUBJECTS/METHODS: Autophagy induction and the expression of autophagy-related proteins were analyzed by LC3 immunofluorescence and western blot, respectively. The role of autophagy and AMP-activated protein kinase (AMPK) was determined by treating NSCLC cells with bafilomycin A1, an autophagy inhibitor, and compound C, an AMPK inhibitor. Cytotoxicity and apoptosis induction were determined by MTT assay, trypan blue exclusion assay, annexin V-propidium iodide (PI) double staining assay, and cell cycle analysis. RESULTS: Morusin increased the formation of LC3 puncta in the cytoplasm and upregulated the expression of autophagy-related 5 (Atg5), Atg12, beclin-1, and LC3II in NSCLC cells, demonstrating that morusin could induce autophagy. Treatment with bafilomycin A1 markedly reduced cell viability but increased proportions of sub-G1 phase cells and annexin V-positive cells in H460 cells. These results indicate that morusin can trigger autophagy in NSCLC cells as a defense mechanism against morusin-induced apoptosis. Furthermore, we found that AMPK and its downstream acetyl-CoA carboxylase (ACC) were phosphorylated, while mammalian target of rapamycin (mTOR) and its downstream p70S6 kinase (p70S6K) were dephosphorylated by morusin. Morusin-induced apoptosis was significantly increased by treatment with compound C in H460 cells. These results suggest that morusin-induced AMPK activation could protect NSCLC cells from apoptosis probably by inducing autophagy. CONCLUSIONS: Our findings suggest that combination treatment with morusin and autophagy inhibitor or AMPK inhibitor might enhance the clinical efficacy of morusin for NSCLC.

Detection of Hydrocarbons Induced by Electron Beam Irradiation of Almond (Prunus amygosalus L.) and Peanut (Arachis hypogaea) (전자선 조사한 아몬드(Prunus amygosalus L.)와 땅콩(Arachis hypogaea)에서 유래한 지방분해산물 분석)

  • Jeong, In Seon;Kim, Jae Sung;Hwang, In Min;Choi, Sung Hwa;Choi, Ji Yeon;Nho, Eun Yeong;Khan, Naeem;Kim, Byung Sook;Kim, Kyong Su
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.20-24
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    • 2013
  • Food irradiation has recently become one of the most successful techniques to preserve food with increased shelf life. This study aims to analyze hydrocarbons in almonds (Prunus amygosalus L.) and peanuts (Arachis hypogaea) induced by electron beam irradiation. The samples were irradiated at 0, 1, 3, 5 and 10 kGy by e-beam and using florisil column chromatography fat, and content was extracted. The induced hydrocarbons were identified using gas chromatography-mass spectrometry (GC/MS). The major hydrocarbons in both irradiated samples were 1,7-hexadecadiene ($C_{16:2}$) and 8-heptadecene ($C_{17:1}$) from oleic acid, 1,7,10-hexadecatriene ($C_{16:3}$) and 6,9-heptadecadiene ($C_{17:2}$) from linoleic acid and 1-tetradecene ($C_{14:1}$) and pentadecane ($C_{15:0}$) from palmitic acid. Concentrations of the hydrocarbons produced by e-beam were found to be depended upon the composition of fatty acid in both almonds and peanuts. The $C_{n-2}$ compound was found to be higher than $C_{n-1}$ compound in oleic acid and palmitic acid, while in case of linoleic acid, $C_{n-1}$compound was higher than $C_{n-2}$ compound. The radiation induced hydrocarbons were detected only in irradiated samples, with 1 kGy or above, and not in the non-irradiated ones. The production of 1,7-hexadecadiene ($C_{16:2}$), 8-heptadecene ($C_{17:1}$), 1,7,10-hexadecatriene ($C_{16:3}$) and 6,9-heptadecadiene ($C_{17:2}$), in high concentration gave enough information to suggest that these may be the possible marker compounds of electron beam irradiation in almonds and peanuts.

Irradiation-induced Volatile Compounds in Irradiated Chicken by P&T Method (P&T법에 의한 닭고기의 방사선 유래 휘발성 조사물질 구명)

  • Kim, Hun;Cho, Woo-Jin;Jung, Yeon-Jung;Lee, Young-Mi;Jeong, Eun-Jeong;Yoo, Young-Jae;Byun, Myung-Woo;Cha, Yong-Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.6
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    • pp.958-964
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    • 2002
  • To select the irradiation-induced marker components from volatile flavor compounds in irradiated chicken, and complement the extraction problems of liquid continuous extraction (LLCE) method, the volatile compounds of irradiated (0,1,3,5 and 10 kGy) chicken were analyzed by Purge and Trap (P&T) and gas chromatography/mass spectrometry (GC/MS) methods. A total of 119 compounds were detected in irradiated chicken, and these compounds were composed mainly of 7 aldehydes,22 ketones,8 alcohols,30 esters,36 hydrocarbons,8 aromatic compounds and 8 miscellaneous compounds. Among these, only 21 compounds were detected in both LLCE and P&T methods, and the 98 other were detected in omly P&T method. Among volatile compounds detected in irradiated chicken, only 3 compounds such as hexene (r=0.96, p<0.01), propanol (r=0.93, p<0.05) and 1,3-bis(1,1-dimethylethyl) benzene (r=0.96, p<0.05) were newly selected as marker compounds in irradiated chicken by P&T method, which showed significant and high positive correlation coefficient in the change of relative concentration according to the increment of irradiation dosage.