• Title/Summary/Keyword: salt sensitive

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OsATG10b, an Autophagosome Component, Is Needed for Cell Survival against Oxidative Stresses in Rice

  • Shin, Jun-Hye;Yoshimoto, Kohki;Ohsumi, Yoshinori;Jeon, Jong-seong;An, Gynheung
    • Molecules and Cells
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    • v.27 no.1
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    • pp.67-74
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    • 2009
  • Autophagy degrades toxic materials and old organelles, and recycles nutrients in eukaryotic cells. Whereas the studies on autophagy have been reported in other eukaryotic cells, its functioning in plants has not been well elucidated. We analyzed the roles of OsATG10 genes, which are autophagy-related. Two rice ATG10 genes - OsATG10a and OsATG10b - share significant sequence homology (about 75%), and were ubiquitously expressed in all organs examined here. GUS assay indicated that OsATG10b was highly expressed in the mesophyll cells and vascular tissue of younger leaves, but its level of expression decreased in older leaves. We identified T-DNA insertional mutants in that gene. Those osatg10b mutants were sensitive to treatments with high salt and methyl viologen (MV). Monodansylcadaverine-staining experiments showed that the number of autophagosomes was significantly decreased in the mutants compared with the WT. Furthermore, the amount of oxidized proteins increased in MV-treated mutant seedlings. These results demonstrate that OsATG10b plays an important role in the survival of rice cells against oxidative stresses.

Isolation and Identification of Lactic Acid Bacteria Inhibiting Gastro-intestinal Pathogenic Bacteria of Domestic Animal. (가축 소화기 병원성 세균을 저해하는 유산균의 분리 및 동정)

  • Lee, Jae-Yeon;Hwang, Kyo-Yeol;Kim, Hyun-Soo;Kim, Geun;Sung, Soo-Il
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.129-134
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    • 2002
  • To isolate probiotic lactic acid bacteria having superior inhibitory activities against animal gastro-intestinal pathogenic bacteria such as Salmonella gallinarum, Staphylococcus aureus and Escherichia coli, 130 strains were initially isolated from the small intestines of Korean native chickens and 7 lactic acid bacteria were finally selected. By using API CHL kit and 16S rRNA sequencing method, the selected lactic acid bacteria were found to be belonged to genus Lactobacillus except BD14 identified as Pediococcus pentosaceus. Especially, Lactobacillus pentosus K34 showed the highest resistancy to both of HCl and bile salt, as well as the highest inhibitory activities against S. gallinarum, S. aureus and E. coli. All the selected strains were sensitive to various antibiotics such as neomycin, erythromycin, cephalosporin, amoxicillin/clavulanic acid, ampicillin, oxytetracycline, but resistant to ciprofloxacin. All the selected strains except BL strain were resistant to colistin and streptomycin, and BD14, BD16, K34 strains were resistant to gentamicin.

Detection of Antistaphylococcal and Toxic Compounds by Biological Assay Systems Developed with a Reporter Staphylococcus aureus Strain Harboring a Heat Inducible Promoter - lacZ Transcriptional Fusion

  • Chanda, Palas Kumar;Ganguly, Tridib;Das, Malabika;Lee, Chia Yen;Luong, Thanh T.;Sau, Subrata
    • BMB Reports
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    • v.40 no.6
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    • pp.936-943
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    • 2007
  • Previously it was reported that promoter of groES-groEL operon of Staphylococcus aureus is induced by various cellwall active antibiotics. In order to exploit the above promoter for identifying novel antistaphylococcal drugs, we have cloned the promoter containing region ($P_g$) of groES-groEL operon of S. aureus Newman and found that the above promoter is induced by sublethal concentrations of many antibiotics including cell-wall active antibiotics. A reporter S. aureus RN4220 strain (designated SAU006) was constructed by inserting the $P_g$-lacZ transcriptional fusion into its chromosome. Agarose-based assay developed with SAU006 shows that $P_g$ in single-copy is also induced distinctly by different classes of antibiotics. Data indicate that ciprofloxacin, rifampicin, ampicillin, and cephalothin are strong inducers, whereas, tetracycline, streptomycin and vancomycin induce the above promoter weakly. Sublethal concentrations of ciprofloxacin and ampicilin even have induced $P_g$ efficiently in microtiter plate grown SAU006. Additional studies show for the first time that above promoter is also induced weakly by arsenate salt and hydrogen peroxide. Taken together, we suggest that our simple and sensitive assay systems with SAU006 could be utilized for screening and detecting not only novel antistaphylococcal compounds but also different toxic chemicals.

Effect of Salinity on Lignin and Hydroxycinnamic Acid Contents in Rice

  • Wi Seung Gon;Kim Jae-Sung;Kim Jin-Hong;Baek Myung Hwa;Yang Dae-Hwa;Lee Myung Chul;Chung Byung Yeoup
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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    • pp.368-372
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    • 2004
  • The lignin contents between IR-29 and Pokkali were not significantly different in the absence of NaCl, but they were slightly increased at 40 mM NaCl. Although lignin contents were not relatively significantly different between salt treated and control plants, the total yields of alkaline nitrobenzene oxidation ranged from 17.4-20.0 mg/g of cell wall residue at 40 mM NaCl were significantly different compared with control plants (11.8-12.2 mg/g). The total amounts of ester-linked hydroxycin-namic acids in IR-29 were decreased from 14.5 to 9.9mg/g, while Pokkali is almost same levels (14.9-15.0 mg/g) under treated and control with 40 mM NaCl. In contrast, the total amounts of ether-linked hydroxycinnamic acids were increased from 9.4 to 13.9 mg/g together with an opposite trend in Pokkali as a decrease 10.9 to 8.8 mg/g under treated and control with 40 mM NaCl. These results revealed that IR-29 is more sensitive in response to 40 mM NaCl in terms of hydroxycinnamic acids than Pokkali.

Effects of Starvation on Lipid Metabolism and Gluconeogenesis in Yak

  • Yu, Xiaoqiang;Peng, Quanhui;Luo, Xiaolin;An, Tianwu;Guan, Jiuqiang;Wang, Zhisheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.11
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    • pp.1593-1600
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    • 2016
  • This research was conducted to investigate the physiological consequences of undernourished yak. Twelve Maiwa yak ($110.3{\pm}5.85kg$) were randomly divided into two groups (baseline and starvation group). The yak of baseline group were slaughtered at day 0, while the other group of yak were kept in shed without feed but allowed free access to water, salt and free movement for 9 days. Blood samples of the starvation group were collected on day 0, 1, 2, 3, 5, 7, 9 and the starved yak were slaughtered after the final blood sample collection. The liver and muscle glycogen of the starvation group decreased (p<0.01), and the lipid content also decreased while the content of moisture and ash increased (p<0.05) both in Longissimus dorsi and liver compared with the baseline group. The plasma insulin and glucose of the starved yak decreased at first and then kept stable but at a relatively lower level during the following days (p<0.01). On the contrary, the non-esterified fatty acids was increased (p<0.01). Beyond our expectation, the ketone bodies of ${\beta}$-hydroxybutyric acid and acetoacetic acid decreased with prolonged starvation (p<0.01). Furthermore, the mRNA expression of lipogenetic enzyme fatty acid synthase and lipoprotein lipase in subcutaneous adipose tissue of starved yak were down-regulated (p<0.01), whereas the mRNA expression of lipolytic enzyme carnitine palmitoyltransferase-1 and hormone sensitive lipase were up-regulated (p<0.01) after 9 days of starvation. The phosphoenolpyruvate carboxykinase and pyruvate carboxylase, responsible for hepatic gluconeogenesis were up-regulated (p<0.01). It was concluded that yak derive energy by gluconeogenesis promotion and fat storage mobilization during starvation but without ketone body accumulation in the plasma.

Improved Purification Process for Cholera Toxin and its Application to the Quantification of Residual Toxin in Cholera Vaccines

  • Jang, Hyun;Kim, Hyo-Seung;Kim, Jeong-Ah;Seo, Jin-Ho;Carbis, Rodney
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.108-112
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    • 2009
  • A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-${\mu}m$ crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-${\mu}m$ permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH 7.0, containing 1.0 M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of $3.1\;EU/{\mu}g$ of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a $G_{M1}$ ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The $G_{M1}$ ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

Detection of porcine encephalomyocarditis virus by in situ hybridization (In situ hybridization에 의한 돼지 뇌심근염 바이러스의 검출)

  • Oh, Sang-hyeon;Park, Nam-yong;Chung, Ci-young;Cho, Kyoung-oh;Lee, Bong-joo;Park, Young-seok;Park, Hyung-seon
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.148-158
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    • 1999
  • The purpose of this study was to establish a rapid, reliable diagnostic method detecting Encephalomyocarditis virus(EMCV) RNA in formalin-fixed, paraffin-embedded tissues of EMCV naturally infected pigs by cDNA probe of EMC $K_3$, the EMCV strain isolated from Korea. Using a biotin-labelled nick translated probe for the cDNA marker. We made up for some defects of radiolabeled method. In sits hybridization(ISH) technique, differently from the other nucleic acid hybridization methods, is able to detect the virus genome specifically in the state of the intact shapes of cells and/or tissues. We succeeded in performing the experiment to detect the EMCV within 1~2 hours using the $MicroProbe^{TM}$ capaillary action system. In this study, we observed highly specific positive signals of red color by staining the paraffin-embedded tissue sections of naturally EMCV-infected pig organs or tissues, including brain, heart, kidney and lacrimal gland with the Fast Red TR salt/Naphtol phosphate chromogen. The results suggested that this ISH method is considered as a highly sensitive and reliable tool for molecular biologic diagnosis of the EMC viral disease.

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Interaction of Apidaecin Ib with Phospholipid Bilayers and its Edwardsiella Species-specific Antimicrobial Activity

  • Seo, Jung-Kil;Go, Hye-Jin;Moon, Ho-Sung;Lee, Min-Jeong;Hong, Yong-Ki;Jeong, Hyun-Do;Nam, Bo-Hye;Park, Tae-Hyun;Park, Nam-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.33 no.1
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    • pp.115-122
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    • 2012
  • Apidaecin Ib had strong antimicrobial activity against several tested Gram-negative bacteria including Escherichia coli, Enterobacter cloacae, and Shigella flexneri (MECs; $0.3-1.5{\mu}g/mL$), but showed no activity against all the tested Gram-positive bacteria including Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus and one yeast, Candida albicans (MECs; > $125{\mu}g/mL$). Interestingly, this peptide showed potent antibacterial activity only against Edwardsiella species (MECs; $0.6-3.6{\mu}g/mL$) among the tested fish pathogenic bacteria through a bacteriostatic process and showed no significant hemolytic activity. Apidaecin Ib took an unordered structure in all environments and also had very weak membrane perturbation activity even at $25{\mu}M$. Anti-Edwardsiella activity of apidaecin Ib is stronger than those of other antimicrobial polypeptides or antibiotics, but its activity is salt-sensitive. These results suggest that apidaecin Ib has Edwardsiella speciesspecific antibacterial activity and could be applied as new preventive or control additives for Edwardsiella species infection in freshwater fish aquaculture.

Development of Dispersive Liquid-Liquid Microextraction Based on Solidification of Floating Organic Drop for the Sensitive Determination of Trace Copper in Water and Beverage Samples by Flame Atomic Absorption Spectrometry

  • Wu, Chunxia;Zhao, Bin;Li, Yingli;Wu, Qiuhua;Wang, Chun;Wang, Zhi
    • Bulletin of the Korean Chemical Society
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    • v.32 no.3
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    • pp.829-835
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    • 2011
  • A dispersive liquid-liquid microextraction based on solidification of floating organic droplet (DLLME-SFO) has been developed as a new approach for the extraction of trace copper in water and beverage samples followed by the determination with flame atomic absorption spectrometry. In the DLLME-SFO, 8-hydroxy quinoline, 1-dodecanol, and methanol were used as chelating agent, extraction solvent and dispersive solvent, respectively. The experimental parameters related to the DLLME-SFO such as the type and volume of the extraction and dispersive solvent, extraction time, sample volume, the concentration of chelating agent and salt addition were investigated and optimized. Under the optimum conditions, the enrichment factor for copper was 122. The method was linear in the range from 0.5 to $300\;ng\;mL^{-1}$ of copper in the samples with a correlation coefficient (r) of 0.9996 and a limit of detection of $0.1\;ng\;mL^{-1}$. The method was applied to the determination of copper in water and beverage samples. The recoveries for the spiked water and beverage samples at the copper concentration levels of 5.0 and $10.0\;ng\;mL^{-1}$ were in the range between 92.0% and 108.0%. The relative standard deviations (RSD) varied from 3.0% to 5.6%.

Octadecyl-Modified Graphene as an Adsorbent for Hollow Fiber Liquid Phase Microextraction of Chlorophenols from Honey

  • Sun, Meng;Cui, Penglei;Ji, Shujing;Tang, Ranxiao;Wu, Qiuhua;Wang, Chun;Wang, Zhi
    • Bulletin of the Korean Chemical Society
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    • v.35 no.4
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    • pp.1011-1015
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    • 2014
  • Octadecyl-modified graphene (graphene-C18) was fabricated and used as adsorbent in hollow fiber liquid phase microextraction (HF-LPME) for the first time. The extraction performance of graphene-C18 reinforced HF-LPME was evaluated using chlorophenols as model analytes. The factors affecting the extraction efficiency, such as extraction time, pH of the sample solution, agitation rate, the concentration of graphene-C18 and salt addition were optimized. After the graphene-C18 reinforced HF-LPME of the chlorophenols from honey sample, the analytes were separated and determined by high-performance liquid chromatography. The linearity was observed in the range of 5.0-200.0 ng $g^{-1}$ for 2-chlorophenol and 3-chlorophenol, and 2.0-200.0 ng $g^{-1}$ for 2,3-dichlorophenol and 3,4-dichlorophenol, respectively. The limits of detection (S/N = 3) of the method were lower than 1.5 ng $g^{-1}$. The recoveries of the method were between 88% and 108%. The method is simple, sensitive and has been resoundingly applied to analysis of chlorophenols in honey samples.