The bifunctional PheA protein, having chorismate mutase and prephenate dehydratase (CMPD) activities, is one of the key regulatory enzymes in the aromatic amino acid biosynthesis in Escherichia coli, and is negatively regulated by an end-product, phenyalanine. Therefore, PheA protein has been thought as useful for protein engineering to utilize mass production of essential amino acid phenylalanine. To obtain feedback resistant PheA protein against phenylalanine, we mutated by using random mutagenesis, extensively screened, and obtained $pheA^{FBR}$ gene encoding a feedback resistant PheA protein. The mutant PheA protein contains substitution of Leu to Phe at the position of 118, displaying that higher affinity (about $290{\mu}M$) for prephenate in comparison with that (about $850{\mu}M$) of wild type PheA protein. Kinetic analysis showed that the saturation curve of $PheA^{FBR}$ against phenyalanine is hyperbolic rather than that of $PheA^{WT}$, which is sigmoidal, indicating that the L118F mutant enzyme has no cooperative effects in prephenate binding in the presence of phenylalanine. In vitro enzymatic assay showed that the mutant protein exhibited increased activity by above 3.5 folds compared to the wild type enzyme. Moreover, L118F mutant protein appeared insensitive to feedback inhibition with keeping 40% of enzymatic activity even in the presence of 10 mM phenylalanine at which the activity of wild type $PheA^{WT}$ was not observed. The substitution of Leu to Phe in CMPD may induce significant conformational change for this enzyme to acquire feedback resistance to end-product of the pathway by modulating kinetic properties.
About the phenomenon of being imaged of everything, the scholars of the humanities who had studied on the simple reason structure in a text have been in a big agony how accept it. Especially, semiologists have studied about this for a long time and the points at issues of Saussure, Peirce as well as Umbeto Eco are more outstanding. Being based upon his philosophic interesting from medieval esthetics to modern semiotics, Eco was very concerned about the field of general esthetics and poputar arts like television and cartoons. He connected the mutual open-relations between 'signifier' and 'signified' debated in Semiotics with the open and vague modern arts and regarding it as a deviation from the custom, intensively studied the film-media. Saussure is a representative figure of semiotics and explained Sign and the character of semiotics as the division into two parts such as signifier/ signified, form/ substance, langue/ parole, synchrony/ diachrony. The triadic semiotics (the theory that Sign is composed of the triadic structure like sign, referent and interpretant) of Peirce put the new item- 'interpretant' in sign and referent to connect them and open the possibility to introduce time in to the Sign. In this paper, I try to analyze a cartoon film in the semiotic structure with the systemic, reasonable and logical approach and analysis as as possible. While the images shown through a film were depended on the romantic and impressional judge in the past, due to semiotics, it' s quite possible to correlate the procedure of symbolization to social coherence so that we analyze the incredible power of images to suck audiences with the systemetic Sign. I accept all ot film-images including a cartoon film as not the simple esthetic arts but a social custom and system, want to serve as a aid to properly understand world and humanbeings and prevent the film-image from being mystic. A cartoon and a cartoon film which were begun with the link of a text and an illustration give shape to all of images such as materials, places and even thoughts with a cartoon icon existed in only a cartoon. A cartoon and a cartoon film simply and exquistely conceptualize the complex and vague attribute of an organic creature and extend them infinetly beyond language. However, it can be exploited as a mysticism to temptate the general public and a faking material. In addition to that, it can distort our world-knowledge engaging a political power and the massive power of mass media. In this paper, being based on semiotics to approach a cartoon film in a scientific and organic system, I conclude that a non-linguistic cartoon expression is entangled with the manifold signs and implies the supplementary meanings just like a regular linguistic expression. It remarks that the iconic images of a cartoon film are composed of the social codes and can be analyzed on grounds of a linguistic system.
In order to characterize ion channels present in tomato roots, microsomes were incorporated into an artificial lipid bilayer arranged for electrophysiological analysis. Of the five different ion channels that could be found, a channel of 450 pS conductance was found most frequently. This channel displayed subconductance states of 450, 257 and 105 pS. All subconductance states showed linear current-voltage relationships. At positive holding potentials, high frequency of transient channel openings was observed; however, at negative potentials, the open times were long and open probability high. Po was 0.83 at -40 mV. When an additional 50 mM $K^+\;or\;Na^+$ was added to the cis side of bilayer, the reversal potentials shifted in the negative direction to near -10 mV. Thus, the 450 pS cation channel selects poorly between $K^+\;and\;Na^+$. In the presence of $100\;{\mu}M$ metal ions, the channel activity was severely inhibited by $La^{3+},\;Ba^{2+},\;and\;Zn^{2+}$, and Po was decreased to 0.2 or even less. However, $Al^{3+}\;and\;Cd^{2+}$ decreased the activity by only 20%. Interestingly, each metal ion showed different kinetics of channel inhibition. While $500\;{\mu}M\;La^{3+}$ inhibited the activities of all subconductance state, 1 mM $Zn^{2+}$ inhibited all except the 105 pS state. $Cd^{2+}$ changed the gating of the channel from a long-opening state to brief transient openings even at negative holding potentials. These data represent that the metal ions may have different binding sites on the channel protein and could be useful modulators and probes to investigate structural characteristics as well as the functional roles of the 450 pS channel on the root physiology.
Iron (hydro)oxides in aqueous environments are primarily formed due to mining activities, and they are known to be typical colloidal particles disturbing surrounding environments. Among them, hematites are widespread in surface environments, and their behavior is controlled by diverse factors in aqueous environments. This study was conducted to elucidate the effect of environmental factors, such as ionic composition and strength, pH, and natural organic matter (NOM) on the behavior of colloidal hematite particles. In particular, two analytical methods, such as dynamic light scattering (DLS) and single-particle ICP-MS (spICP-MS), were compared to quantify and characterize the behavior of colloidal hematites. According to the variation of ionic composition and strength, the aggregation/dispersion characteristics of the hematite particles were affected as a result of the change in the thickness of the diffuse double layer as well as the total force of electrostatic repulsion and van der Walls attraction. Besides, the more dispersed the particles were, the farther away the aqueous pH was from their point of zero charge (PZC). The results indicate that the electrostatic and steric (structural) stabilization of the particles was enhanced by the functional groups of the natural organic matter, such as carboxyl and phenolic, as the NOM coated the surface of colloidal hematite particles in aqueous environments. Furthermore, such coating effects seemed to increase with decreasing molar mass of NOM. On the contrary, these stabilization (dispersion) effects of NOM were much more diminished by divalent cations such as Ca2+ than monovalent ones (Na+), and it could be attributed to the fact that the former acted as bridges much more strongly between the NOM-coated hematite particles than the latter because of the relatively larger ionic potential of the former. Consequently, it was quantitatively confirmed that the behavior of colloidal hematites in aqueous environments was significantly affected by diverse factors, such as ionic composition and strength, pH, and NOM. Among them, the NOM seemed to be the primary and dominant one controlling the behavior of hematite colloids. Meanwhile, the results of the comparative study on DLS and spICPMS suggest that the analyses combining both methods are likely to improve the effectiveness on the quantitative characterization of colloidal behavior in aqueous environments because they showed different strengths: the main advantage of the DLS method is the speed and ease of the operation, while the outstanding merit of the spICP-MS are to consider the shape of particles and the type of aggregation.
Jung, Jong Hwa;Cho, Sung Bae;Kim, Jineun;Kim, Jae Sang;Lee, Shim Sung
Analytical Science and Technology
/
v.6
no.1
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pp.29-37
/
1993
Some new podands containing phenyl(B), benzyl(Bz), pyridine(Py), quinoline(Q) and naphthalene(Np) as end-groups, and oxygen(O) and sulfur(S) in ether chains as donor atoms have been synthesized. The univalent cation binding characteristics of these podands have been studied by NMR titration and solvent extraction. By NMR titration we have found that the most of podands form 1:1 complexes with $Ag^+$ ion. Especially, the substituted sulfur atoms in ether chains show the effects to enhance the stabilities. We also carried out the extractions of univalent cation picrates including alkaline metal, $Ag^+$, $Tl^+$ and $NH_4{^-}$ ions from aqueous to chloroform layer by using these podands. We found that the extractabilities of $Ag^+$ ion with the quinoline-containing podands such as, $Q_2O_4$, $Q_2O_5$ and $BQO_5$ were 86.8, 86.6 and 48.0% respectively, but the naphthalene-containing podands such as, $Np_2O_4$ and $Np_2O_5$ extracted quite small amount. Otherwise, in cases of $Bz_2O_3S_2$(89.4%), $B_2O_2S_2$(96.8%), $B_2O_3S_2$(58.9%), $Py_2O_2S_2$(58.8%), $Py_2O_3S_2$(42.1%), and $B_2O_4S$(15.0%), interestingly, $Bz_2O_3S_2$ which have sulfur atoms and benzyl groups showed the highest extraction selectivity for $Ag^+$ ion. This result seems due to not only the strong interaction of $Ag^+$ ion with sulfur donors according to the HSAB theory, but also the effective ${\pi}-{\pi}$ stacking interaction between two aromatic end-groups which is enhanced by the flexible methylene spacing group in benzyl groups instead of phenyl groups. The extraction coefficients gave the similar tendency as the extractabilities and the stabilities. From these results, it could be concluded that the predominant factor affected to extraction coefficients is the stabilities, which are strongly influenced by the structures of podands.
Imicyafos which is a nematicide for controlling root-knot nematodes has been registered in the Republic of Korea in 2012, and the maximum residue limits of imicyafos are set to watermelon and korean melon as each 0.05 mg/kg. Extremely reliable and sensitive analytical method is required for ensuring food safety on imicyafos residues in agricultural commodities. Imicyafos residues in samples were extracted with acetone, partitioned with hexane and dichloromethane, and then purified with florisil. The purified samples were analyzed by HPLC-UVD and confirmed with LC-MS. Linear range was between 0.1~5 mg/kg with the correlation coefficient ($r^2$) 0.99997. Average recoveries of imicyafos ranged from 77.0 to 115.4% at the spiked levels of 0.02 and 0.05 mg/kg with the relative standard deviations of 2.2~9.6%. Limit of detection and quantification were 0.005 and 0.02 mg/kg, respectively. An inter-laboratory study was conducted to validate the determination method in depth, and the results were satisfactory. All of the validation results revealed that the developed analytical method in this study is relevant for imicyafos determination in agricultural commodities and will be used as an official analytical method.
Many of the protective functions of saliva can be attributed to the biological, physical, structural, and rheological characteristics of salivary glycoproteins. Therefore, the development of ideal saliva substitutes requires understanding of the rheological as well as biological properties of human saliva. In the present study, we investigated the changes of salivary enzymatic activities by saliva substitutes and compared viscosity of saliva substitutes with human saliva. Five kinds of saliva substitutes such as Moi-Stir, Stoppers4, MouthKote, Saliva Orthana, and SNU were used. Lysozyme activity was determined by the turbidimetric method. Peroxidase activity was determined with an NbsSCN assay. $\alpha$-Amylase activity was determined using a chromogenic substrate, 2-chloro-p-nitrophenol linked with maltotriose. The pH values of saliva substitutes were measured and their viscosity values were measured with a cone-and-plate digital viscometer at six different shear rates. Various types of saliva substitutes affected the activities of salivary enzymes in different ways. Stoppers4 enhanced the enzymatic activities of hen egg-white lysozyme, bovine lactoperoxidase (bLP), and $\alpha$-amylase. Saliva Orthana and SNU inhibited bLP activity and enhanced $\alpha$-amylase activity. MouthKote inhibited $\alpha$-amylase activity. Moi-Stir inhibited the enzymatic activities of bLP and $\alpha$-amylase. The pH values were very different according to the types of saliva substitutes. Stoppers4, MouthKote, and Saliva Orthana showed lower values of viscosity at low shear rates and higher values of viscosity at high shear rates compared with unstimulated and stimulated whole saliva. Moi-Stir and SNU displayed much higher values of viscosity than those of natural whole saliva. Collectively, our results indicate that each saliva substitute has its own biological and rheological characteristics. Each saliva substitute affects the enzymatic activity of salivary enzyme and finally oral health in different ways.
Journal of the Korean Society of Marine Environment & Safety
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v.24
no.5
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pp.628-636
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2018
Because large buoys are mainly made of steel, they are heavy and vulnerable to corrosion by sea water. This makes buoy installation and maintenance difficult. Moreover, vessel collision accidents with buoys and damage to vessels due to the material of buoys (e.g., steel) are reported every year. Recently, light buoys adopting eco-friendly and lightweight materials have come into the spotlight in order to solve the previously-mentioned problems. In Korea, a new lightweight buoy with a 7-Nautical Mile lantern adopting expanded polypropylene (EPP) and aluminum to create a buoyant body and tower structure, respectively, was developed in 2017. When these light buoys are operated in the ocean, the visibility and angle of light from the lantern installed on the light buoys changes, which may cause them to function improperly. Therefore, research on the performance of light buoys is needed since the weight distribution and motion characteristics of these new buoys differ from conventional models. In this study, stability estimation and motion analyses for newly-developed buoys under various environmental conditions considering a mooring line were carried out using ANSYS AQWA. Numerical simulations for the estimation of wind and current loads were performed using commercial CFD software, Siemens STAR-CCM+, to increase the accuracy of motion analysis. By comparing the estimated maximum significant motions of the light buoys, it was found that waves and currents were more influential in the motion of the buoys. And, the estimated motions of the buoys became larger as the sea state became worser, which might be the reason that the peak frequencies of the wave spectra got closer to those of the buoys.
We prepared and characterized $Cu(In_{1-x}Ga_x)Se_2$(CIGS) films using a elemental co-evaporation method for absorbing layer of high efficiency thin film solar cells. The CIGS films deposited on a soda-lime glass exhibited low resistivity because of higher carrier concentration. Na was accumulated at the CIGS surface and the 0 and Se were also accumulated at the surface, suggesting that oxidation is a driving force of Na accumulation. The structure of CIGS film was modified or a secondary phase was formed in the Cu-poor CIGS bulk films probably due to the incorporation of Na into Cu vacancy sites. As the Ga/(In+Ga) ratio increased, the diffraction peaks of $Cu(In_{1-x}Ga_x)Se_2$ films were shifted to larger angle and splitted, and the grain size of $Cu_{0.91}(In_{1-x}Ga_x)Se_2$ films became smaller. All $Cu_{0.91}(In_{1-x}Ga_x)Se_2$ films showed the p-type conductivity regardless of the Ga/(In+Ga) ratio. Ag/n-ZnO/i-ZnO/CdS/$Cu_{0.91}(In_{0.7}Ga_{0.3})Se_2$/Mo solar cells were fabricated. The currently best efficiency in this study was 14.48% for $0.18cm^2$ area ($V_{oc}=581.5mV,\;J_{sc}=34.88mA$, F.F=0.714).
Microbial lethal value and nutrient retention of sous vide processed spinach were evaluated with mathematical model prediction and experimental trial for different package sizes and pasteurization temperatures. The package size covers 500 g, 1 kg and 2 kg, while the pasteurization temperature includes 80, 90 and 97$^{\circ}C$. The basic process scheme consists of filling blanched spinach into barrier plastic film pouch, sealing under vacuum, pasteurization in hot water with over pressure and final cooling to 3$^{\circ}C$. Pasteurization condition was designed based on attainment of 6 decimal inactivation of Listeria monocytogenes at geometric center of the pouch package by heating cycle, which was determined by general method. Heat penetration property of the package and thermal destruction kinetics were combined to estimate the retention of ascorbic acid and chlorophyll. Smaller packages with shorter pasteurization time gave better nutrient retention, physical and chemical qualities. Larger package size was estimated and confirmed experimentally to give higher pasteurization value at center, lower ascorbic acid and chlorophyll contents caused by longer heat process time. Lower pasteurization temperature with longer process time was predicted to give lower pasteurization value at center and lower ascorbic acid, while chlorophyll content was affected little by the temperature. Experimental trial showed better retention of ascorbic acid and chlorophyll for smaller package and higher pasteurization temperature with shorter heating time. The beneficial effect of smaller package and higher pasteurization temperature was also observed in texture, color retention and drip production.
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