• Journal of Life Science
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• v.28 no.9
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• pp.1030-1041
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• 2018

Abiotic stresses limit the growth and productivity of plants. Cellular adaptation to abiotic stresses requires coordinated regulation in gene expression directed by complex mechanisms. This study used the activation tagging system to identify novel salt stress-responsive genes. The study selected 9 activation tagging lines that showed salt stress-tolerant phenotypes during their germination stages. Thermal asymmetric interlaced-PCR (TAIL-PCR) was used to identify the T-DNA tagging sites on the Arabidopsis genome in selected activation tagging lines, including AT7508, AT7512, AT7527, AT7544, AT7548, and AT7556. RT-PCR analysis showed that ClpC2/HSP93-III (At3g48870), plant thionin family (At2g20605), anti-muellerian hormone type-2 receptor (At3g50685), vacuolar iron transporter family protein (At4g27870), and microtubule-associated protein (At5g16730) were activated in AT7508, AT7512, AT7527, AT7544, and AT7556, respectively. Interestingly, in AT7548, both the genes adjacent to the T-DNA insertion site were activated: Arabinogalactan protein 13 (AGP13) (At4g26320) and F-box/RNI-like/FBD-like domains-containing protein (At4g26340). All of the seven genes were newly identified as salt stress-responsive genes from this study. Among them, the expression of ClpC2/HSP93-III, AGP13, F-box/RNI-like/FBD-like domains-containing protein gene, and microtubule-associated protein gene were increased under salt-stress condition. In addition, AT7508, AT7527, and AT7544 were more tolerant to salt stress than wild type at seedling development stage, functionally validating the screening results of the activation tagging lines. Taken together, our results demonstrate that the activation tagging system is useful for identifying novel stress-responsive genes.

• Natural Product Sciences
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• v.22 no.3
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• pp.201-208
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• 2016

Here in this study, we investigated the lifespan-extending effect and underlying mechanism of methanolic extract of Moringa olelifa leaves (MML) using Caenorhabditis elegans (C. elegans) model system. To define the longevity properties of MML we conducted lifespan assay and MML showed significant increase in lifespan under normal culture condition. In addition, MML elevated stress tolerance of C. elegans to endure against thermal, oxidative and osmotic stress conditions. Our data also revealed that increased activities of antioxidant enzymes and expressions of stress resistance proteins were attributed to MML-mediated enhanced stress resistance. We further investigated the involvement of MML on the aging-related factors such as growth, food intake, fertility, and motility. Interestingly, MML significantly reduced growth and egg-laying, suggesting these factors were closely linked with MML-mediated longevity. We also observed the movement of aged worms to estimate the effects of MML on the health span. Herein, MML efficiently elevated motility of aged worms, indicating MML may affect health span as well as lifespan. Our genetic analysis using knockout mutants showed that lifespan-extension activity of MML was interconnected with several genes such as skn-1, sir-2.1, daf-2, age-1 and daf-16. Based on these results, we could conclude that MML prolongs the lifespan of worms via activation of SKN-1 and SIR-2.1 and inhibition of insulin/IGF pathway, followed by DAF-16 activation.

• Journal of Microbiology and Biotechnology
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• v.17 no.8
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• pp.1344-1352
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• 2007

An opportunistic human pathogen, Pseudomonas aeruginosa, contains the major catalase KatA, which is required to cope with oxidative and osmotic stresses. As an attempt to uncover the $H_2O_2$-dependent regulatory mechanism delineating katA gene expression, four prototrophic $H_2O_2$-sensitive mutants were isolated from about 1,500 TnphoA mutant clones of P. aeruginosa strain PA14. Arbitrary PCR and direct cloning of the transposon insertion sites revealed that one insertion is located within the katA coding region and two are within the coding region of oxyR, which is responsible for transcriptional activation of several antioxidant enzyme genes in response to oxidative challenges. The fourth insertion was within PA3815 (IscR), which encodes a homolog of the Escherichia coli iron-sulfur assembly regulator, IscR. The levels of catalase and SOD activities were significantly reduced in the iscR mutant, but not in the oxyR mutant, during the normal planktonic culture conditions. These results suggest that both IscR and OxyR are required for the optimal resistance to $H_2O_2$, which involves the expression of multiple antioxidant enzymes including KatA.

• Journal of Ginseng Research
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• v.9 no.1
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• pp.119-127
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• 1985

The effect of ginseng saponin on acid phosphatase (AP) activity in liver Iysosomes was investigated and the mechanism by which ginseng saponin may function on the integrity of Iysosomes was discussed. The experimental results obtained are summarized as follows; 1, A very marked increase in the AP activity was observed in the supernatant of hypotonic medium, as compared with that of isotonic medium, indicating that the hypoosmotic shock per so results in activation through osmotic Iysis of particles. 2. Ginseng saponin had no effect on the activity of AP if once released from Iysosomes when Iysed in the hypotonic medium, suggesting that ginseng saponin has no effect on the enzyme molecules per se. 3. The AP activity in isotonic medium suspensions was decreased at the concentrations of 10-6, 10-5 and 10-4% of ginseng saponin, but increased at 10-2 and 10-1%. It's suggested that ginseng saponin enhances the integrity of Iysosomes at 10-6, 10-5 and 10-4%, but decreases it at 10-2 and 10-1%. 4. Suspending particles in distilled water resulted in no correlation of AP activity with treatment with ginseng saponin. 5, The AP activity was decreased in the presence of ATP, showing the possible significance of ATP as a Iysosomal stabilizer and the possibility that ginseng saponin may affect a membrane bound ATPase system by which Iysosomal AP release may be controlled.

• Journal of Life Science
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• v.6 no.3
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• pp.198-203
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• 1996

A thermolabile alkaline phosphatase has been purified through steps of osmotic shock, ammonium sulfate salting-out, and DAEA-cellulose chromatography from the cultured broth of the marine Vibrio sp. M-96 strain. The optimal temperature for the enzyme activity was 35$\circ$C. The optimal pH was pH11.0, and the range of pHstability was pH10.4 to 12.0. Thermal inactivation occured within 6 mintes at 60$\circ$C. The enzyme was considerably inactivated by 0.1mM concentrations of Hg$^{2+}$, Ni$^{2+}$ and Zn$^{2+}$, whereas activated up to 234% by 1mM of Mn$^{2+}$. The activation energy and deactivation energy by the Arrhenius equation were 4.02 Kcal/mol and 9.098 Kcal/mol, respectively. The Km and Vmax values of the enzyme for p-introphenylphosphate were found to be 0.0465mM and 0.001334mM/min, respectively. Active form of the enzyme had a molecular weight of 57,000 dalton determined by the Sephadex G-200 gel filtration method.

• Transactions of the Korean hydrogen and new energy society
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• v.32 no.1
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• pp.1-10
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• 2021

Water electrolysis technology, which generates hydrogen using renewable energy resources, has recently attracted great attention. Especially, the polymer electrolyte membrane water electrolysis system has several advantages over other water electrolysis technologies, such as high efficiency, low operating temperature, and optimal operating point. Since research that analyzes performance characteristics using test bench have high cost and long test time, however, model based approach is very important. Therefore, in this study, a system model for water electrolysis dynamics of a polymer electrolyte membrane was developed based on MATLAB/Simulink®. The water electrolysis system developed in this study can take into account the heat and mass transfer characteristics in the cell with the load variation. In particular, the performance of the system according to the stack temperature control can be analyzed and evaluated. As a result, the developed water electrolysis system can analyze water pump dynamics and hydrogen generation according to temperature dynamics by reflecting the dynamics of temperature.

• Nutrition Research and Practice
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• v.11 no.5
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• pp.430-434
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• 2017

BACKGROUND/OBJECTIVE: Chronic hyperglycemia induces oxidative stress via accumulation of reactive oxygen species (ROS) and contributes to diabetic complications. Hyperglycemia induces mitochondrial superoxide anion production through the increased activity of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. This study aimed to determine whether fisetin and luteolin treatments suppress the oxidative stress by modulating the expression of sirtuins (SIRTs) and forkhead box O3a (FOXO3a) under hyperglycemic conditions in human monocytes. MATERIALS/METHODS: Human monocytic cells (THP-1) were cultured under osmotic control (14.5 mmol/L mannitol), normoglycemic (NG, 5.5 mmol/L glucose), or hyperglycemic (HG, 20 mmol/L glucose) conditions, in the absence or presence of fisetin and luteolin for 48 h. To determine the effect of fisetin and luteolin treatments on high glucose-induced oxidative stress, western blotting and intracellular staining were performed. RESULTS: Hyperglycemic conditions increased the ROS production, as compared to normoglycemic condition. However, fisetin and luteolin treatments inhibited ROS production under hyperglycemia. To obtain further insight into ROS production in hyperglycemic conditions, evaluation of p47phox expression revealed that fisetin and luteolin treatments inhibited p47phox expression under hyperglycemic conditions. Conversely, the expression levels of SIRT1, SIRT3, SIRT6, and FOXO3a were decreased under high glucose conditions compared to normal glucose conditions, but exposure to fisetin and luteolin induced the expression of SIRT1, SIRT3, SIRT6, and FOXO3a. The above findings suggest that fisetin and luteolin inhibited high glucose-induced ROS production in monocytes through the activation of SIRTs and FOXO3a. CONCLUSIONS: The results of our study supports current researches that state fisetin and luteolin as potential agents for the development of novel strategies for diabetes.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.1
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• pp.9-14
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• 2009

We cultured canine kidney(MDCK) cells stably expressing aquaporin-2(AQP2) on collagen-coated permeable membrane filters and examined the effect of extracellular ATP on arginine vasopressin(AVP)-stimulated fluid transport and cAMP production. Exposure of cell monolayers to basolateral AVP resulted in stimulation of apical to basolateral net fluid transport driven by osmotic gradient which was formed by addition of 500 mM mannitol to basolateral bathing solution. Pre-exposure of the basolateral surface of cell monolayers to ATP(100 ${\mu}M$) for 30 min significantly inhibited the AVP-stimulated net fluid transport. In these cells, AVP-stimulated cAMP production was suppressed as well. Profile of the effects of different nucleotides suggested that the $P2Y_2$ receptor is involved in the action of ATP. ATP inhibited the effect of isoproterenol as well, but not that of forskolin to stimulate cAMP production. The inhibitory effect of ATP on AVP-stimulated fluid movement was attenuated by a protein kinase C inhibitor, calphostin C or pertussis toxin. These results suggest that prolonged activation of the P2 receptors inhibits AVP-stimulated fluid transport and cAMP responses in AQP2 transfected MDCK cells. Depressed responsiveness of the adenylyl cyclase by PKC-mediated modification of the pertussis-toxin sensitive $G_i$ protein seems to be the underlyihng mechanism.

• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.391-396
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• 1996

The Gram-negative marine bacterium Marinomonas vaga, which requires 0.5 M NaCl concentration for optimal growth, is slightly halophilic. The growth of M vaga was highly resistant to the proton conductor, carbonyl cyanide m-chlorophenylhydrazone (CCCP) under alkaline pH conditions (pH 8.5) but very sensitive to CCCP under acidic pH conditions (pH 6.5). These results suggest that the respiratory chain-linked NADH oxidase system of M. vaga may lead to generation of a $Na^{+}$ electrochemical gradient. In order to examine the existence of $Na^{+}$-stimulated NADH oxidase in M. vaga, membrane fractions were prepared by the osmotic lysis method. The membrane-bound NADH oxidase oxidized both NADH and deamino-NADH as substrates and required $Na^{+}$ for maximum activity. The maximum activity of NADH oxidase was obtained at about pH 8.5 in the presence of 0.2 M NaCl. The site of $Na^{+}$-dependent activation in the NADH oxidase system was at the NADH:quinone oxidoreductase segment. The NADH oxidase and NADH:quinone oxidoreductase were very sensitive to the respiratory chain inhibitor, 2-heptyl-4-hydroxyquinoline-N-oxide (HQNO) in the presence of 0.2 M NaCl but highly resistant to another respiratory inhibitor, rotenone. Based on these findings, we conclude that M. vaga possesses the $Na^{+}$-dependent NADH:quinone oxidoreductase that may function as an electrogenic $Na^{+}$ pump.

• Journal of The Korean Society of Grassland and Forage Science
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• v.34 no.1
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• pp.15-20
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• 2014

To investigate lignification process and its physiological significance under water-deficit condition, the responses of peroxidases, polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL) in relation to leaf water status to the short term of water deficit treatment in the leaves with different maturities in forage rape were measured. The significant decrease in relative water content (RWC) and leaf osmotic potential (${\Psi}{\pi}$) were apparent after 5 d of water-deficit treatment. The activity of guaiacol peroxidase (GPOD), ascorbate peroxidase (APOD), coniferyl alcohol peroxidase (CPOD), and syringaldazine peroxidase (SPOD) was depressed especially in middle and old leaves when compared with that of control leaves. On the other hand, in young leaves, a significant increase in CPOD (+34%) and SPOD (+24%) activity as affected by water-deficit treatment was apparent. The activation of PAL and PPO was observed in middle and old leaves for PAL and in young and middle leaves for PPO. These results suggest that peroxidases in middle and old leaves did not involve in lignification under mild water-deficit stress, whereas CPOD and SPOD in young leaves participate in lignification by a coordination with PAL and PPO to incorporate phenol and lignin into the cell walls.