• Title/Summary/Keyword: Rice cells

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Inhibitory Effect of the Ethanol Extract of a Rice Bran Mixture Comprising Angelica gigas, Cnidium officinale, Artemisia princeps, and Camellia sinensis on Brucella abortus Uptake by Professional and Nonprofessional Phagocytes

  • Hop, Huynh Tan;Arayan, Lauren Togonon;Reyes, Alisha Wehdnesday Bernardo;Huy, Tran Xuan Ngoc;Baek, Eun Jin;Min, WonGi;Lee, Hu Jang;Lee, Chun Hee;Rhee, Man Hee;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1885-1891
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    • 2017
  • In this study, we evaluated the inhibitory effect of a rice bran mixture extract (RBE) on Brucella abortus pathogenesis in professional (RAW 264.7) and nonprofessional (HeLa) phagocytes. We fermented the rice bran mixture and then extracted it with 50% ethanol followed by gas chromatography-mass spectrometry to identify the components in RBE. Our results clearly showed that RBE caused a significant reduction in the adherence of B. abortus in both cell lines. Furthermore, analysis of phagocytic signaling proteins by western blot assay revealed that RBE pretreatment resulted in inhibition of phosphorylation of JNK, ERK, and p38, leading to decline of internalization compared with the controls. Additionally, the intensity of F-actin observed by fluorescence microscopy and FACS was remarkably reduced in RBE-pretreated cells compared with control cells. However, the intracellular replication of B. abortus within phagocytes was not affected by RBE. Taken together, these findings suggest that the phagocytic receptor blocking and suppressive effects of RBE on the MAPK-linked phagocytic signaling pathway could negatively affect the invasion of B. abortus into phagocytes.

Suppression of Inflammatory Responses by Black Rice Extract in RAW 264.7 Macrophage Cells via Downregulation of NF-kB and AP-1 Signaling Pathways

  • Limtrakul, Pornngarm;Yodkeeree, Supachai;Pitchakarn, Pornsiri;Punfa, Wanisa
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.10
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    • pp.4277-4283
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    • 2015
  • Anthocyanin, a phenolic compound, has been reported to have an anti-inflammatory effect against lipopolysaccharide (LPS) induced changes in immune cells. However, little is known about the molecular mechanisms underlying its anti-inflammatory effects. Few research studies have concerned the anti-inflammation properties of colored rice extract as a functional material. Therefore, the purpose of this study was to examine anti-inflammatory effects of the polar fraction of black rice whole grain extracts (BR-WG-P) that features a high anthocyanin content. Our results showed that BR-WG-P significantly inhibited LPS-induced pro-inflammatory mediators, including production of NO and expression of iNOS and COX-2. In addition, secretion of pro-inflammatory cytokines including TNF-${\alpha}$ and IL-6 was also significantly inhibited. Moreover, BR-WG-P and anthocyanin inhibited NF-kB and AP-1 translocation into the nucleus. BR-WG-P also decreased the phosphorylation of ERK, p38 and JNK in a dose dependent manner. These results suggested that BR-WG-P might suppress LPS-induced inflammation via the inhibition of the MAPK signaling pathway leading to decrease of NF-kB and AP-1 translocation. All of these results indicate that BR-WG-P exhibits therapeutic potential associated with the anthocyanin content in the extract for treating inflammatory diseases associated with cancer.

A PAS-Containing Histidine Kinase is Required for Conidiation, Appressorium Formation, and Disease Development in the Rice Blast Fungus, Magnaporthe oryzae

  • Shin, Jong-Hwan;Gumilang, Adiyantara;Kim, Moon-Jong;Han, Joon-Hee;Kim, Kyoung Su
    • Mycobiology
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    • v.47 no.4
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    • pp.473-482
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    • 2019
  • Rice blast disease, caused by the ascomycete fungus Magnaporthe oryzae, is one of the most important diseases in rice production. PAS (period circadian protein, aryl hydrocarbon receptor nuclear translocator protein, single-minded protein) domains are known to be involved in signal transduction pathways, but their functional roles have not been well studied in fungi. In this study, targeted gene deletion was carried out to investigate the functional roles of the PAS-containing gene MoPAS1 (MGG_02665) in M. oryzae. The deletion mutant ΔMopas1 exhibited easily wettable mycelia, reduced conidiation, and defects in appressorium formation and disease development compared to the wild type and complemented transformant. Exogenous cAMP restored appressorium formation in ΔMopas1, but the shape of the restored appressorium was irregular, indicating that MoPAS1 is involved in sensing the hydrophobic surface. To examine the expression and localization of MoPAS1 in M. oryzae during appressorium development and plant infection, we constructed a MoPAS1:GFP fusion construct. MoPAS1:GFP was observed in conidia and germ tubes at 0 and 2 h post-infection (hpi) on hydrophobic cover slips. By 8 hpi, most of the GFP signal was observed in the appressoria. During invasive growth in host cells, MoPAS1:GFP was found to be fully expressed in not only the appressoria but also invasive hyphae, suggesting that MoPAS may contribute to disease development in host cells. These results expand our knowledge of the roles of PAS-containing regulatory genes in the plant-pathogenic fungus M. oryzae.

Production of Green Fluorescent Protein (GFP) from Transgenic Rice Cell Suspension Culture (형질전환된 벼세포배양에서 green fluorescent protein (GFP) 생산)

  • Lee, Jae-Hwa
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.293-297
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    • 2007
  • Green fluorescent protein (GFP) is an attractive reporter for bioprocess monitoring. A fluorescence-based method was developed to quantify GFP levels in transgenic plants and protein extracts. In this study, GFP was produced and secreted from suspension cells derived from transgenic rice. The RAmy3E promoter placed before the GFP gene controlled by sugars such as sucrose. The effects of sucrose concentration on the secretion of GFP and total protein into the medium were investigated in batch suspension culture. It was possible, therefore, to induce the expression of the GFP by removing sucrose from the cultured media or by allowing the rice suspension cells to deplete sucrose catabolically. The dry cell weight (7.06 g/L) and GFP level were detected as highest at 12%, 3% sucrose after 20 day culture, respectively. However secreted GFP fluorescence at the other sucrose concentrations (6%, 12%, 18% and 24%) were a little amount in media.

Resveratrol-Enriched Rice Down-Regulates Melanin Synthesis in UVB-Induced Guinea Pigs Epidermal Skin Tissue

  • Lee, Taek Hwan;Seo, Jae Ok;Do, Moon Ho;Ji, Eunhee;Baek, So-Hyeon;Kim, Sun Yeou
    • Biomolecules & Therapeutics
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    • v.22 no.5
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    • pp.431-437
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    • 2014
  • Synthetic compounds that are used in the clinic to regulate skin hyperpigmentation, such as arbutin, hydroquinone, and kojic acid, are only moderately effective. But, their use is limited by side effects. As part of an effort to overcome the limitations, we developed resveratrol-enriched rice (RR) using genetic engineering technique. Each of resveratrol and rice has been reported to produce anti-melanogenic effects. Therefore, we hypothesized that RR would show more anti-melanogenic effects than those of resveratrol or rice alone. Anti-melanogenic effect of RR was done by using melan-a mouse melanocytes. The depigmenting efficacy was then observed following topical application of the RR to UVB-stimulated hyperpigmented dorsal skin of guinea pigs. Treatment with RR extract resulted a $21.4{\pm}0.7%$ decrease in tyrosinase expression at melan-a cells. Colorimetric analysis showed a significantly lower depigmenting value by day 9 following treatment with RR in UVB-irradiated guinea pigs the dorsal skin (p<0.01), indicating that RR produced a depigmentation effect. By staining with Fontana-Masson stain, we found that the RR-treated group had more effect histopathologically in epidermal melanin production than resveratrol or rice alone-treated group. RR was associated with reduction in the levels of microphthalmia-associated transcription factor (MITF), and downregulation of tyrosinase and tyrosinase-related protein (TRP-2) expression, leading to inhibit epidermal melanin production by western blot analysis. This study suggests that the resveratrol-enriched rice may be a promising candidate in regulating skin pigmentation with UVB exposure.

An Effective Defensive Response in Thai Aromatic Rice Varieties(Oryza sativa L. spp. indica) to Salinity

  • Cha-um, Suriyan;Vejchasarn, Phanchita;Kirdmanee, Chalermpol
    • Journal of Crop Science and Biotechnology
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    • v.10 no.4
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    • pp.257-264
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    • 2007
  • Rice is one of the world's staple crops and is a major source of carbohydrate. Rice is exported from several countries, providing a major source of income. There are many documents reporting that rice is a salt-sensitive crop in its developmental stages. The objective of this investigation is to evaluate the effective salt-tolerance defense mechanisms in aromatic rice varieties. Pathumthani 1(PT1), Jasmine(KDML105), and Homjan(HJ) aromatic rice varieties were chosen as plant materials. Rice seedlings photoautotrophically grown in-vitro were treated with 0, 85, 171, 256, 342, and 427 mM NaCl in the media. Data, including sodium ion$(Na^+)$ and potassium ion$(K^+)$ accumulation, osmolarity, chlorophyll pigment concentration, and the fresh and dry weights of seedlings were collected after salt-treatment for 5 days. $Na^+$ in salt-stressed seedlings gradually accumulated, while $K^+$ decreased, especially in the 342-427 mM NaCl salt treatments. The $Na^+$ accumulation in both salt-stressed root and leaf tissues was positively related to osmolarity, leading to chlorophyll degradation. In the case of the different rice varieties, the results showed that the HJ variety was identified as being salt-tolerant, maintaining root and shoot osmolarities as well as pigment stabilization when exposed to salt stress or $Na^+$ enrichment in the cells. On the other hand, PT1 and KDML105 varieties were classified as salt-sensitive, determined by chlorophyll degradation using Hierarchical cluster analysis. In conclusion, the HJ-salt tolerant variety should be further utilized as a parental line or genetic resource in breeding programs because of the osmoregulation defensive response to salt-stress.

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The Calmodulin-Binding Transcription Factor OsCBT Suppresses Defense Responses to Pathogens in Rice

  • Koo, Sung Cheol;Choi, Man Soo;Chun, Hyun Jin;Shin, Dong Bum;Park, Bong Soo;Kim, Yul Ho;Park, Hyang-Mi;Seo, Hak Soo;Song, Jong Tae;Kang, Kyu Young;Yun, Dae-Jin;Chung, Woo Sik;Cho, Moo Je;Kim, Min Chul
    • Molecules and Cells
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    • v.27 no.5
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    • pp.563-570
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    • 2009
  • We previously isolated the OsCBT gene, which encodes a calmodulin (CaM)-binding protein, from a rice expression library constructed from fungal elicitor-treated rice suspension cells. In order to understand the function of OsCBT in rice, we isolated and characterized a T-DNA insertion mutant allele named oscbt-1. The oscbt-1 mutant exhibits reduced levels of OsCBT transcripts and no significant morphological changes compared to wild-type plant although the growth of the mutant is stunted. However, oscbt-1 mutants showed significant resistance to two major rice pathogens. The growth of the rice blast fungus Magnaporthe grisea, as well as the bacterial pathogen Xanthomonas oryzae pv. oryzae was significantly suppressed in oscbt-1 plants. Histochemical analysis indicated that the hypersensitive-response was induced in the oscbt-1 mutant in response to compatible strains of fungal pathogens. OsCBT expression was induced upon challenge with fungal elicitor. We also observed significant increase in the level of pathogenesis-related genes in the oscbt-1 mutant even under pathogen-free condition. Taken together, the results support an idea that OsCBT might act as a negative regulator on plant defense.

Physiological Functionality and Cytotoxic Effect of Korean Traditional Noble Wine, Samhaeju, and Commercial Rice Wine on Various Tumor Cell Lines (삼해주와 시판 곡주의 생리 기능성 및 세포 독성 효과)

  • Lim, Chae-Lan;Son, Hee-Jin;Cho, In-Young;Kim, Gye-Won;Choi, Soo-Jin;Kim, In-Sun;Han, Kee-Young;Choi, Jin-Young;Noh, Bong-Soo
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.687-693
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    • 2009
  • This study was conducted to investigate the antioxidant activity, fibrinolytic activity and cytotoxic effect of Korean traditional noble rice wine made using different methods (A-C) and commercial rice wine (D-H) on various tumor cell lines. The antioxidant activity of rice wine was measured by DPPH (2,2-dipicryl-1-picrylhydrazyl), ABTS [2,2'-azinobis(3-ethylbenzthiazoline-6-sulphonic acid)] and NO (nitric oxide) radical scavenging assay. In this study, Samhaeju showed the greatest fibrinolytic activity of 13-17U and exhibited the highest antioxidant activity. Among the different Samhaeju, the sample prepared using method C had the highest antioxidant activity. The cytotoxic effect of rice wine were also examined against the human cancer cell line (A549 cells and HeLa cells) based on the results of a WST-1 assay and morphological changes. Rice wine induced the inhibition of cell proliferation and morphological changes in tumor cell lines in a concentration-dependent manner, with Samhaeju diluted 10 fold having the strongest effect on these factors. These findings suggest that Korean rice wine has antioxidant activity and cytotoxic effect, and that these factors are influenced by the method of preparation.

Exclusion of Na+ and ClIons by the central parenchyma in leaf sheaths of rice and the involvement of lamina joint

  • Neang, Sarin;Kano-Nakata, Mana;Yamauchi, Akira;Itani, Tomio;Maekawa, Masahiko;Mitsuya, Shiro
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.237-237
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    • 2017
  • Rice is highly sensitive to salt stress especially in its early growth stage, which thus is one of the major constraints in rice production. In rice plants, salt sensitivity is associated with the accumulation of $Na^+$ in the shoots, especially in the photosynthetic tissues. High salt concentrations in soil cause high $Na^+$ and $Cl^-$ transport to the shoot and preferential accumulation of those ions in older leaves, which decreases $K^+$ in the shoot, photosynthetic activity and grain yield. Salt exclusion capacity at the leaf sheath is therefore considered to be one of the main mechanisms of salt tolerance. In addition, it is suspected that the lamina joint might be involved in the salt transport from leaf sheath to leaf blade. This research aims to determine if leaf sheaths of rice exclude a large amount of $Na^+$ only or other ions such as $K^+$, $Ca^{2+}$, $Mg^{2+}$, and $Cl^-$ as well, to identify tissues in the leaf sheath, which accumulate $Na^+$, and to examine if the lamina joint is involved in the salt exclusion by the leaf sheath. The rice seedlings of salt tolerant genotype FL478 and salt sensitive genotype IR29 were independently treated with NaCl, KCl, $MgCl_2$ and $CaCl_2$, and Taichung 65 and its near-isogenic liguleless line (T65lg) were treated with NaCl. Then, the content of $Na^+$, $K^+$, $Ca^{2+}$, $Mg^{2+}$, and $Cl^-$ ions and their specific location were determined using Atomic Absorption Spectrometer, Ion Chromatograph, and Energy Dispersive X-ray Spectroscopy. Results showed that leaf sheaths of FL478 and IR29 accumulated a large amount of $Na^+$, $K^+$, $Ca^{2+}$, $Mg^{2+}$, and $Cl^-$ ons, and thus excluded them from leaf blades when treated with high concentration of each salt. When treated with NaCl, the highest $Na^+$ concentration was found in the basal part of leaf sheaths of both cultivars. Moreover, energy-dispersive X-ray spectroscopy revealed that the central parenchyma cells of the leaf sheath were the site where most Na, Cl, and K were retained under salinity in the salt tolerant genotype FL478. Also, the concentration of $Na^+$, $K^+$ and $Cl^-$ ions in leaf sheaths and leaf blades was comparable between T65 and T65lg, indicating that the lamina joint may not be involved in the exclusion of $Na^+$, $Cl^-$ and $K^+$ by the leaf sheath from the leaf blade under salinity. Therefore, we conclude that the central parenchyma cells of basal part of leaf sheath are the site that plays a physiological role to exclude $Na^+$ in the shoots of rice without the involvement of the lamina joint.

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Dealcoholized Korean Rice Wine (Makgeolli) Exerts Potent Anti-Tumor Effect in AGS Human Gastric Adenocarcinoma Cells and Tumor Xenograft Mice

  • Shin, Eun Ju;Kim, Sung Hee;Kim, Jae Ho;Ha, Jaeho;Hwang, Jin-Taek
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1485-1492
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    • 2015
  • Makgeolli is a traditional wine in Korea and has been traditionally believed to exhibit health benefits. However, the inhibitory effect of dealcoholized makgeolli (MK) on cancer has never been investigated scientifically. In this study, MK exhibited an anti-angiogenic effect by inhibiting tube formation in human umbilical vein endothelial cells, without cytotoxicity. Treatment with MK reduced the proliferation of AGS human gastric adenocarcinoma cells in a dose-dependent manner and increased the sub-G1 population. Next, we evaluated whether MK could induce apoptosis in AGS cells by using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay or Annexin V method. Treatment with MK at 500 and 1,000 μg/ml increased the number of TUNEL-positive AGS cells. Under the same conditions, MK-treated (500 and 1,000 μg/ml) cells showed significant induction of early or late apoptosis, compared with untreated cells (no induction). In addition, MK also induced phosphatase and tensin homolog (PTEN) expression in AGS cells. However, p53 expression in AGS cells was not changed by MK treatment. Furthermore, MK at 500 mg/kg·d reduced the tumor size and volume in AGS tumor xenografts. Taken together, MK may be useful for the prevention of cancer cell growth.