Xanthomonas oryzae pv. oryzae (Xoo) produces a putative effector, XoAvrBs2. We expressed XoAvrBs2 homologously in Xoo with a TAP-tag at the C-terminus to enable quantitative analysis of protein expression and secretion. Addition of rice leaf extracts from both Xoo-sensitive and Xoo-resistant rice cultivars to the Xoo cells induced expression of the XoAvrBs2 gene at the transcriptional and translational levels, and also stimulated a remarkable amount of XoAvrBs2 secretion into the medium. In a T3SS-defective Xoo mutant strain, secretion of the TAPtagged XoAvrBs2 was blocked. Thus, we elucidated the transcriptional and translational expressions of the XoAvrBs2 gene in Xoo was induced in vitro by the interaction with rice and the induced secretion of XoAvrBs2 was T3SSdependent. It is the first report to measure the homologous expression and secretion of XoAvrBs2 in vitro by rice leaf extract. Our system for the quantitative analysis of effector protein expression and secretion could be generally used for the study of host-pathogen interactions.
A 1,989-bp genomic region encoding nickel resistance genes was isolated from Legionella pneumophila, a pathogen for legionellosis. From a sequencing and computer analysis, the region was found to harbor two structural genes, a nreB-like protein gene (1,149 bp) and a nreA-like protein gene (270 bp), in a row. Both genes exhibited a significant degree of similarity to the corresponding genes from Synechocystis sp. PCC6803 ($54\%$ amino acid sequence identity) and Achromobacter xylosoxidans 31A ($76\%$). The gene was successfully expressed in E. coli MG1655 and conferred a nickel resistance of up to 5 mM in an LB medium and 3 mM in a TMS medium including gluconate as the sole carbon source. E. coli harboring the nickel resistance gene also exhibited a substantial resistance to cobalt, yet no resistance to cadmium or zinc. Since the extracellular concentration of nickel remained constant during the whole period of cultivation, it was confirmed that the nickel resistance was provided by an efflux system like the $Ni^2+$permease (nrsD) of Synechocystis sp. strain PCC6803. Since polyphosphate (poly-P) is known as a global regulator for gene expression as well as a potential virulence factor in E. coli, the nickel resistance of a ppk mutant of E. coli MG 1655 harboring the nickel resistance gene from L. pneumophila was compared with that of its parental strain. The nickel resistance was significantly attenuated by ppk inactivation, which was more pronounced in an LB medium than in a TMS medium.
Journal of the Society of Cosmetic Scientists of Korea
/
v.35
no.2
/
pp.159-169
/
2009
In this study, the antibacterial activity, antioxidative effects, inhibitory effects on tyrosinase, inhibitory effects on elastase, and components of Quercus acutissima Carruth leaf extracts were investigated. MIC values of ethyl acetate fraction from Q. acutissima Carruth leaf on P. acnes, S. aureus, P. ovale, and E. coli were 0.13 %, 0.25 %, 0.13 % and 0.25 %, respectively. The results showed that the antibacterial activity of the ethyl acetate fraction was the highest in the S. aureus, P. acnes, and P. ovale. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) of extract/fractions of Q. acutissima Carruth. leaf was in the order: 50 % ethanol extract (12.13 ${\mu}g/mL$) < ethyl acetate fraction (7.07 ${\mu}g/mL$) < deglycosylated flavonoid aglycone fraction (6.20 ${\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some Q. acutissima Carruth leaf extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was 50 % ethanol extract ($OSC_{50}$, 1.81 ${\mu}g/mL$) < ethyl acetate fraction (1.70 ${\mu}g/mL$) < deglycosylated flavonoid aglycone fraction (0.70 ${\mu}g/mL$). Deglycosylated flavonoid aglycone fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Q. acutissima Carruth leaf on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Q. acutissima Carruth leaf extracts suppressed photohemolysis in a concentration dependent manner, particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect (${\tau}50$, 220.00 min at 25 ${\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Q. acutissima Carruth leaf extracts, showed 3 bands (QA 1, QA2 and QA3) on TLC. TLC chromatogram of ethyl acetate fraction of Q. Carruth. leaf extract revealed 4 bands (QA 1 ${\sim}$ QA 4), Among them, kaempferol (QA 1), quercetin (QA 2), and gallic acid (QA 3) were identified. The inhibitory effect ($IC_{50}$) of aglycone fraction on tyrosinase was 65.7 ${\mu}g/mL$. The inhibitory effect ($IC_{50}$) of aglycone fraction on elastase was 24.50 ${\mu}g/mL$. These results indicate that extract/fractions of Q. acutissima Carruth. can functionized as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. Extract/fractions of Q. acutissima Corruth can be applicable to new functional cosmetics for antioxidant, antiaging, antibacterial activity.
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.196-196
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2017
Oilseed crop Camelina (Camelina sativa L.) is a suitable for biodiesel production that has high adaptability under low-nutrient condition like marginal land and requires low-input cost for cultivation. Enhanced abiotic stress tolerance of Camelina is very important for oil production under the wide range of different climate. CsRCI2s (Rare Cold Inducible 2) are related proteins in various abiotic stresses that predicted to localized at plasma membrane (PM) and endoplasmic reticulum (ER). These proteins are consist of eight-family that can be divided into tail (CsRCI2D/E/F/G) and no-tail (CsRCI2A/B/E/H) type of C-terminal. However, it is still less understood the function of C-terminal tail. In this study, CsRCI2D/H genes were cloned through gateway cloning system that used pCB302-3 as destination vector. And we used agrobacterium-mediated transformation system for generation of overexpression (OX) transformants. Overexpression of target gene was confirmed using RT-PCR and segregation ratio on selection media. We analyzed physiological response in media and soil under abiotic stresses using CsRCI2D and CsRCI2H overexpression plant. To compare abiotic stresses tolerance, wild type and CsRCI2D/H OX line seeds were sown on agar plate treated with various NaCl and mannitol concentration for 7 days. In the test of growth rate under abiotic stress on media, CsRCI2H OX line showed similar to NaCl and mannitol stress. In the other hand, CsRCI2D OX line showed to be improved stress tolerance that especially increased in 200mM NaCl but was similar on mannitol media. In greenhouse, WT and CsRCI2D/H OX lines for physiological analysis and productivity under abiotic stresses were treated 100, 150, 200mM NaCl. Then it was measured various parameters such as leaf width and length, plant height, total seed weight, flower number, seed number. CsRCI2H OX line in greenhouse did not show any changes in physiological parameters but CsRCI2D OX line was improved both physiological response and productivity under NaCl stress. Among physiological parameters of CsRCI2D OX line under NaCl stress, leaf length and width were observed shorter than WT but it were slightly longer than WT in 200mM NaCl stress. Furthermore, total seed weight of CsRCI2D OX line under stress displayed to decrease than WT in normal condition, but it was gradually raised with increasing NaCl stress then more than WT relatively. These results suggested CsRCI2D might be contribute to improve abiotic stress tolerance. However, function of CsRCI2H is need to more detail study. In conclusion, overexpression of CsRCI2s family can generate various environmental stress tolerance plant and may improve crop productivity for bio-energy production.
TSAHC [4'-(p-toluenesulfonylamido)-4-hydroxychalcone] is a promising antitumorigenic chalcone compound, especially against TM4SF5 (four-transmembrane L6 family member 5)-mediated hepatocarcinoma. We evaluated the potential of TSAHC to inhibit the catalytic activities of nine cytochrome P450 isoforms and of P-glycoprotein (P-gp). The abilities of TSAHC to inhibit phenacetin O-deethylation (CYP1A2), coumarin 6-hydroxylation (CYP2A6), bupropion hydroxylation (CYP2B6), amodiaquine N-deethylation (CYP2C8), diclofenac 4-hydroxylation (CYP2C9), omeprazole 5-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), and midazolam 1'-hydroxylation (CYP3A) were tested using human liver microsomes. The P-gp inhibitory effect of TSAHC was assessed by [$^3H$]digoxin accumulation in the LLCPK1-MDR1 cell system. TSAHC strongly inhibited CYP2C8, CYP2C9, and CYP2C19 isoform activities with $K_i$ values of 0.81, 0.076, and $3.45{\mu}M$, respectively. It also enhanced digoxin accumulation in a dose-dependent manner in the LLCPK1-MDR1 cells. These findings indicate that TSAHC has the potential to inhibit CYP2C isoforms and P-gp activities in vitro. TSAHC might be used as a nonspecific inhibitor of CYP2C isoforms based on its negligible inhibitory effect on other P450 isoforms such as CYP1A2, CYP2A6, CYP2B6, CYP2D6, CYP2E1, and CYP3A.
Peptidylprolyl cis-trans isomerase (PPlase) catalyzes the cis-trans isomerization of prolyl peptide and facilitates the folding of cellular proteins and peptides. PPlase consists of two distinct immunophilins, each specifically binding to the immunosupressive drug cyclosporin A (CsA) or FK506, respectively. A PPlase was isolated and partially purified from porcine spleen. The molecular weight of porcine spleen PPlase was determined to be ~14,000 on the basis of SDS-PAGE. The purified enzyme was strongly inhibited by FK506, but not by CsA. The inhibition constant and the true concentration of enzyme preparations were determined by active site titration using the tight binding inhibitor FK506: $K_{i}=18.7$ nM and $E_{t}=172$ nM. The equilibrium ratio of conformer. [cis]/[trans], of prolyl peptide substrates (N-Suc-Ala-Xaa-Pro-Phe-p-NA) in anhydrous trifluoroethanol/LiCl solvent system varied from 0.24 to 0.85 depending on the nature of Xaa. Overall. in this solvent-salt system, the populations of the cis conformer of substrates in equilibrium are higher than in an aqueous solution so that the substantial error caused by high background absorption can be reduced. The reactivities of porcine spleen PPlase are shown to be highly sensitive to changes in the structure of substrates. Thus, $k_{cat}/K_m$ value for the most reactive substrate (Xaa Leu) is $4.007+10^{6}M^{1}s^{1}$ and, is 2,636 fold higher than that for the least reactive peptide substrate tested, Xaa=Glu.
Species distribution modeling is one of the most effective habitat analysis methods for wildlife conservation. This study was for evaluating the suitability of species distribution to distance between forest patches in Seoul city using tits. We analyzed the distribution of the four species of tits: varied tit (Parus varius), marsh tit (P. palustris), great tit (P. major) and coal tit (P. ater), using the landscape indexes and connectivity indexes, and compared the resulting suitability indexes from 100m to 1,000m. As factors affecting to the distribution of tits, we calculated landscape indices by separating them into intra-patch indices (i.e. logged patch area (PA), area-weighted mean patch shape index (PSI), tree rate (TR)) and inter-patch indices (i.e. patch degree (PD), patch betweenness (PB), difference probability of connectivity (DPC)), to analyze the internal properties of the patches and their connectivity by tits occurrence data using logistic regression modeling. The models were evaluated by AICc (Akaike Information Criteria with a correction for finite sample sizes) and AUC (Area Under Curve of ROC). The results of AICc and AUC showed DPC, PA, PSI, and TR were important factors of the habitat models for great tit and marsh tit at the level of distance 500~800m. In contrast, habitat models for coal tit and varied tit, which are known as forest interior species, reflected PA, PSI, and TR as intra-patch indices rather than connectivity. These mean that coal tit and varied tit are more likely to find a large circular forest patch than a small and long-shaped forest patch, which are higher rate of forest. Therefore, different strategies are required in order to enhance the habitats of the forest birds, tits, in a region that has fragmented forest patches such as Seoul city. It is important to manage forest interior areas for coal tit and varied tit, which are known as forest interior species and to manage not only forest interior areas but also connectivity of the forest patches in the threshold distance for great tit and marsh tit as adapted species to the urban ecosystem for sustainable ecosystem management.
Gould, Andrew;Jung, Youn Kil;Hwang, Kyu-Ha;Dong, Subo;Albrow, Michael D.;Chung, Sun-Ju;Han, Cheongho;Ryu, Yoon-Hyun;Shin, In-Gu;Shvartzvald, Yossi;Yang, Hongjing;Yee, Jennifer C.;Zang, Weicheng;Cha, Sang-Mok;Kim, Dong-Jin;Kim, Seung-Lee;Lee, Chung-Uk;Lee, Dong-Joo;Lee, Yongseok;Park, Byeong-Gon;Pogge, Richard W.
Journal of The Korean Astronomical Society
/
v.55
no.5
/
pp.173-194
/
2022
We complete the survey for finite-source/point-lens (FSPL) giant-source events in 2016-2019 KMTNet microlensing data. The 30 FSPL events show a clear gap in Einstein radius, 9 𝜇as < 𝜃E < 26 𝜇as, which is consistent with the gap in Einstein timescales near tE ~ 0.5 days found by Mróz et al. (2017) in an independent sample of point-source/point-lens (PSPL) events. We demonstrate that the two surveys are consistent. We estimate that the 4 events below this gap are due to a power-law distribution of free-floating planet candidates (FFPs) dNFFP/d log M = (0.4 ± 0.2) (M/38 M⊕)-p/star, with 0.9 ≲ p ≲ 1.2. There are substantially more FFPs than known bound planets, implying that the bound planet power-law index 𝛾 = 0.6 is likely shaped by the ejection process at least as much as by formation. The mass density per decade of FFPs in the Solar neighborhood is of the same order as that of 'Oumuamua-like objects. In particular, if we assume that 'Oumuamua is part of the same process that ejected the FFPs to very wide or unbound orbits, the power-law index is p = 0.89 ± 0.06. If the Solar System's endowment of Neptune-mass objects in Neptune-like orbits is typical, which is consistent with the results of Poleski et al. (2021), then these could account for a substantial fraction of the FFPs in the Neptune-mass range.
Kim, Soo-Kyoung;Kim, Cheol-Jin;Yoon, Je-Yong;Lee, Joon-Hee
Microbiology and Biotechnology Letters
/
v.39
no.1
/
pp.29-36
/
2011
Pseudomonas aeruginosa is an opportunistic pathogen that causes various infections on urinary track, cornea, respiratory track, and burn wound site, and mainly relies on quorum sensing (QS) for its virulence. To control the infectivity of P. aeruginosa, we previously synthesized the structural analogues of a major QS signal, N-3-oxododecanoyl homoserine lactone (3OC12-HSL) to use as a QS inhibitor. Two of them (5b and 5f) had been confirmed to have an inhibitory effect on LasR, a major QS signal receptor of P. aeruginosa in the screening by the recombinant Escherichia coli reporter. To further evaluate these compounds, we tested their efficacy to control the QS and virulence of P. aeruginosa. Unlike the result from E. coli reporter, both 5b and 5f failed to affect the LasR activity in P. aeruginosa, but instead they selectively affected the activity of QscR, another 3OC12-HSL receptor of P. aeruginosa. Interestingly, their effect on QscR was complex and opposite to what we obtained with E. coli system. Both 5b and 5f enhanced the QscR activity at the low concentration range (< 10 ${\mu}m$), but high concentration of 5f (${\approx}$1 mM) strongly inhibited QscR. While 5b and 5f didn't affect the production of proteases, the key virulence factor, they significantly reduced the biofilm formation that is important in mediating chronic infections. Especially, 5f inhibited the initial attachment of P. aeruginosa, rather than the biofilm maturation. Based on our results, we suggest that 5f can be applied for an anti-biofilm agent without increasing virulence of P. aeruginosa.
Korean Journal of Air-Conditioning and Refrigeration Engineering
/
v.9
no.2
/
pp.171-179
/
1997
During the day time in summer, peak of air conditing load, and electric power management system lies under overloaded condition. The reason is the enlarged peak load value of electric power caused by increased air-cooling load in summer. To prevent load concentration during day time and overloaded condition of power management system, some energy storage methods are suggested. One of these methods is ice storage system. Water has some good properties as P.C.M.(Phase Chang Material) : Its melting point is the range of required operation temperature. It has large specific latent heat and is chemically stable compared to other organic or inorganic substances. It is cheap and easy to treat. This study represents experimental results of heat transfer characteristics of P.C.M. under the outward melting process in a vertical cylinder. We experimented with twelve combinations of conditions, i.e., three different inlet temperatures($7^{\circ}C,\;4^{\circ}C\;and\;1^{\circ}C$), two working fluid directions(upward and downward), and two aspect ratios, H/R(4 and 2). At the inlet temperature of $7^{\circ}C$ and $4^{\circ}C$, there was temperature stagnation region where the temperature of P.C.M. remains constant at $4^{\circ}C$ regardless of aspect ratio and direction of working fluid. This temperature stagnation occurs as the water, at its maximum density, flows down to the lower region. The phase change interface formed bell-shaped curve as the melting process continued. With a new set of conditions(4H/R, inlet temperature $4^{\circ}C$ and $1^{\circ}C$, downward/upwerd inlet direction), the movement of phase change interface was faster when the working flued inlet direction was downward. With the same set of conditions, melting rate and total melting energy were larger when the working fluid inlet direction was downward. The results were reversed when the other sets of conditions were applied.
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