Agmatine iminohydrolase (EC 3.5.3.12) catalyzes the hydrolysis of agmatine into putrescine. The enzyme seems to be one of the critical enzymes in putrescine biosynthesis. The enzyme was purified to homogeneity from Panax ginseng C.A. Meyer by combined method of ammonium sulfate 1 fractionation, DEAR anion exchange column, hydroxyapatite column and agmatine carboxyhexyl Sepharose 4B affinity column. The molecular weight estimated by native pore gadient polyacrylamide gel electrophoresis was 71, 000 Dalton, while that estimated by SDS-PAGE was 70, 000 Dalton, indicating a monomeric enzyme. The optimal pH and temperature were 9.0 and 37$^{\circ}C$, respectively. The Km and 1 Vmax for agmatine were 8.3 mM and 14.4 nmole/hr, respectively. Heat stability of this enzyme was high. The enzyme was observed to be inhibited by polyamines such as putrescine, cadaverine, spermidine and spermine. Especially, putrescine was a potent inhibitor of the purified enzyme. These results suggest that polyamines could be important in growth regulation of Panax ginseng C.A. Meyer.
Journal of the Korean Society for Aviation and Aeronautics
/
v.20
no.2
/
pp.32-38
/
2012
The United Nations (UN) has tried to make international agreement to restrict artificial greenhouse gas emissions and the UN has concluded the UN Framework Convention Climate Change (UNFCCC) and the Kyoto protocol. Moreover, in 2012, the European Union announced that they will enforce the Directive 2008/101/EC. Therefore, after 2012, aircraft carriers that depart or arrive from EU will follow that regulation. For these reasons, Korea should prepare systematic and effective policy to reduce greenhouse gas emission from aviation activities. The purpose of this study is to find out effective measures to reduce greenhouse gas emission from aviation activities through research by countries all over the world. Here are the 4 measures to reduce greenhouse gas emission from Aviation activities that were found through research UK and Japan's policies. First, Korea should implement aggressive incentive policies. Providing proper incentive can attract voluntary participation of aircraft carriers to reduce greenhouse gas emission. Second, the government should adopt environmental tax on use of fossil fuels. Third, Korea should adopt the greenhouse gas Emissions Trading Scheme (ETS). Lastly, the Korea government should pull in with the international community to establish world-wide environmental policies.
This study was conducted to find a system for screening organic resources with 16 species, 62 samples which were selected to randomizing point from city, province and industrial areas in the whole country. Content of organic matters were $65.3%{\sim}98.0%$ in all samples so that they were largely over than 60%, raw material regulation of compost. Concentrations of total nitrogen and total phosphorus were $0.7{\sim}4.8%\;and\;0.8{\sim}5.0$, they could look for effect of the nitrogen and phosphorus supply as a raw material of compost. In case of 8 elements concentrations of heavy metal, they were too high to use as raw materials of compost which were over to regulation limit in Cu, Cr, Ni, and As from fiber industry, Ni from food company and leather industry, and the others are adapt to limit levels. HEM contents fro the highest to $113mg\;kg^{-1}$ from liber industry and PAHs content were the highest to $3,462ug\;kg^{-1}$ from paper-mill manufacture. Distribution of PAHs concentiations were naphthalene>phenanthrene>pyrene>fluoroanthene>acenaphthene. $Microtox{(R)}\;EC_{50}$ values for bioassay were pharmaceutical company>paper-mill manufacture>industrial area sewage sludge>fiber industry>urban sewage sludge>metropolitan sewage sludge. HEM between Zn, Cu, and Ni was significant at the 99% and between Cd was significant at the 95%, Microtox between Hg and BEM significant at the 95%.
This paper reviews and criticizes the EU Case of C-452/13, Germanwings GmbH v. Ronny Henning. Under this case, Ronny Henning later sued Lufthansa's budget carrier Germanwings after it refused to pay him 250 euros compensation for a delay he said totalled more than three hours. Germanwings, however, maintained his flight had arrived only two hours and 58 minutes behind schedule. In those circumstances, the following question to the European Court of Justice (ECJ) for a preliminary ruling: What time is relevant for the term time of arrival used in Articles 2, 5 and 7 of Regulation [No 261/2004]: (a) the time that the aircraft lands on the runway (touchdown); (b) the time that the aircraft reaches its parking position and the parking brakes are engaged or the chocks have been applied (in-block time); (c) the time that the aircraft door is opened; (d) a time defined by the parties in the context of party autonomy? ECJ says that the situation of passengers on a flight does not change substantially when their aircraft touches down on the runway at the destination airport, when that aircraft reaches its parking position and the parking brakes are engaged or when the chocks are applied, as the passengers continue to be subject, in the enclosed space in which they are sitting, to various constraints. Therefore, it is only when the passengers are permitted to leave the aircraft and the order is given to that effect to open the doors of the aircraft that the passengers may in principle resume their normal activities without being subject to those constraints. ECJ rules that it is apparent that Articles 2, 5 and 7 of Regulation No 261/2004 must be interpreted as meaning that the concept of 'arrival time', which is used to determine the length of the delay to which passengers on a flight have been subject, corresponds to the time at which at least one of the doors of the aircraft is opened, the assumption being that, at that moment, the passengers are permitted to leave the aircraft.
Kim, Dong-Uk;Kim, Ji-Hoon;Kim, Sung-Mi;Kwon, Ky-Beom
The Korean Journal of Air & Space Law and Policy
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v.32
no.1
/
pp.225-285
/
2017
In regard to the regulations related to the RPA(Remotely Piloted Aircraft), which is sometimes called in other countries as UA(Unmanned Aircraft), ICAO stipulates the regulations in the 'RPAS manual (2015)' in detail based on the 'Chicago Convention' in 1944, and enacts provisions for the Rules of UAS or RPAS. Other contries stipulates them such as the Federal Airline Rules (14 CFR), Public Law (112-95) in the United States, the Air Transport Act, Air Transport Order, Air Transport Authorization Order (through revision in "Regulations to operating Rules on unmanned aerial System") based on EASA Regulation (EC) No.216/2008 in the case of unmanned aircaft under 150kg in Germany, and Civil Aviation Act (CAA 1998), Civil Aviation Act 101 (CASR Part 101) in Australia. Commonly, these laws exclude the model aircraft for leisure purpose and require pilots on the ground, not onboard aricraft, capable of controlling RPA. The laws also require that all managements necessary to operate RPA and pilots safely and efficiently under the structure of the unmanned aircraft system within the scope of the regulations. Each country classifies the RPA as an aircraft less than 25kg. Australia and Germany further break down the RPA at a lower weight. ICAO stipulates all general aviation operations, including commercial operation, in accordance with Annex 6 of the Chicago Convention, and it also applies to RPAs operations. However, passenger transportation using RPAs is excluded. If the operational scope of the RPAs includes the airspace of another country, the special permission of the relevant country shall be required 7 days before the flight date with detail flight plan submitted. In accordance with Federal Aviation Regulation 107 in the United States, a small non-leisure RPA may be operated within line-of-sight of a responsible navigator or observer during the day in the speed range up to 161 km/hr (87 knots) and to the height up to 122 m (400 ft) from surface or water. RPA must yield flight path to other aircraft, and is prohibited to load dangerous materials or to operate more than two RPAs at the same time. In Germany, the regulations on UAS except for leisure and sports provide duty to avoidance of airborne collisions and other provisions related to ground safety and individual privacy. Although commercial UAS of 5 kg or less can be freely operated without approval by relaxing the existing regulatory requirements, all the UAS regardless of the weight must be operated below an altitude of 100 meters with continuous monitoring and pilot control. Australia was the first country to regulate unmanned aircraft in 2001, and its regulations have impacts on the unmanned aircraft laws of ICAO, FAA, and EASA. In order to improve the utiliity of unmanned aircraft which is considered to be low risk, the regulation conditions were relaxed through the revision in 2016 by adding the concept "Excluded RPA". In the case of excluded RPA, it can be operated without special permission even for commercial purpose. Furthermore, disscussions on a new standard manual is being conducted for further flexibility of the current regulations.
Objective: Follicle-stimulating hormone (FSH) is the central hormone involved in mammalian reproduction, maturation at puberty, and gamete production that mediates its function by control of follicle growth and function. The present study investigated the mutations involved in the regulation of FSH receptor (FSHR) activation. Methods: We analyzed seven naturally-occurring mutations that were previously reported in human FSHR (hFSHR), in the context of equine FSHR (eFSHR); these include one constitutively activation variant, one allelic variant, and five inactivating variants. These mutations were introduced into wild-type eFSHR (eFSHR-wt) sequence to generate mutants that were designated as eFSHR-D566G, -A306T, -A189V, -N191I, -R572C, -A574V, and -R633H. Mutants were transfected into PathHunter EA-parental CHO-K1 cells expressing β-arrestin. The biological function of mutants was analyzed by quantitating cAMP accumulation in cells incubated with increasing concentrations of FSH. Results: Cells expressing eFSHR-D566G exhibited an 8.6-fold increase in basal cAMP response, as compared to that in eFSHR-wt. The allelic variation mutant eFSHR-A306T was not found to affect the basal cAMP response or half maximal effective concentration (EC50) levels. On the other hand, eFSHR-D566G and eFSHR-A306T displayed a 1.5- and 1.4-fold increase in the maximal response, respectively. Signal transduction was found to be completely impaired in case of the inactivating mutants eFSHR-A189V, -R572C, and -A574V. When compared with eFSHR-wt, eFSHR-N191I displayed a 5.4-fold decrease in the EC50 levels (3,910 ng/mL) and a 2.3-fold decrease in the maximal response. In contrast, cells expressing eFSHR-R633H displayed in a similar manner to that of the cells expressing the eFSHR-wt on signal transduction and maximal response. Conclusion: The activating mutant eFSHR-D566G greatly enhanced the signal transduction in response to FSH, in the absence of agonist treatment. We suggest that the state of activation of the eFSHR can modulate its basal cAMP accumulation.
Embryonic stem (ES) cells can self-renew maintaining the undifferentiated state. Self-renewal requires many factors such as OCT4, SOX2, and NANOG. It is previously known that OCT4 and SOX2 can bind to NANOG promoter and support Nanog gene expression in mouse ES cells by the detailed studies using the mouse Nanog promoter. Here, we constructed serial deletion mutant promoter-reporter constructs to investigate the human Nanog gene promoter in detail. The highest promoter activity was obtained in the 0.6 kb (-253/+365) promoter-reporter construct which includes the binding sites of OCT4 and SOX2. To further confirm contribution of OCT4 and SOX2 in Nanog gene expression, we introduced site- directed mutation(s) in the OCT4 and/or SOX2 binding sites of the human Nanog promoter 0.6 kb (-253/+365) and checked the influence of the mutation on the promoter activity using human EC cell line NCCIT. Mutation either in OCT4 binding site or SOX2 binding site significantly reduced the activity of Nanog promoter which directly confirmed that OCT4 and SOX2 binding is essential in human Nanog gene expression.
Journal of the Society of Cosmetic Scientists of Korea
/
v.23
no.2
/
pp.23-32
/
1997
Melanin pigmentation in human skin is a major defense mechanism against ultraviolet light of the sun. Tyrosinase(EC 1.14.18.1) plays a key role in the biosynthesis of ultraviolet of melanin. This is why much researches have been focused on its regulation in controlling the epidermal melanization. We have found that the water-extract of Banha(Pinelliae ternate B.), an oriental medicinal plant, has no tyrosinase inhibitory activity, but does inhibit the melanin biolsynthesis in B16 mouse melanin cells. We also found that Banha extract lowers the tyrosinase activity in cultured cells. To elucidate the action mechanism of Banha extract we have investigated its effect on the tyrosinase mRNA level using reverse transcription-polymerase chain reaction technique. It was revealed that Banha extract reduced the tyrosinase mRNA level in dose dependent manner; when B16 mouse melanoma cells were cultured with 2mg/ml and 5mg/ml of Banha extract, there were 20% and 44% decrease in tyrosinase mRNA level, respectively. These data suggest that the Banha extract exerts its melanogenic inhibitory effect through the transcriptional regulation of tyrosinase mRNA.
$\gamma$-Glutamyl transpeptidase (GGT, EC 2.3.2.2.) catalyzes the transfer of the $\gamma$-glutamyl moiety from $\gamma$-glutamyl containing ompounds, notably glutathione (GSH), to acceptor amino acids and peptides. A second gene (GGTII) encoding GGT was previously isolated and characterized from the fission yeast Schizosaccharomyces pombe. In the present work, the GGTII-lacZ fusion gene was constructed and used to study the transcriptional regulation of the S. pombe GGTII gene. The synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene was significantly enhanced by NO-generating SNP and hydrogen peroxide in the wild type yeast cells. The GGTII mRNA level was increased in the wild-type S. pombe cells treated with SNP. However, the induction by SNP was abolished in the Pap1-negative S. pombe cells, implying that the induction by SNP of GGTII is mediated by Pap1. Fermentable carbon sources, such as glucose (at low concentrations), lactose and sucrose, as a sole carbon source, enhanced the synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene in wild type KP1 cells but not in Pap1-negative cells. Glycerol, a non-fermentable carbon source, was also able to induce the synthesis of $\beta$-galactosidase from the fusion gene, but other non-fermentable carbon sources such as acetate and ethanol were not. Transcriptional induction of the GGTII gene by fermentable carbon sources was also confirmed by increased GGTII mRNA levels in the yeast cells grown with them. Nitrogen starvation was also able to induce the synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene in a Pap1-dependent manner. On the basis of the results, it is concluded that the S. pombe GGTII gene is regulated by oxidative and metabolic stress.
The enzyme responsible for the synthesis of nitric oxide (NO) from L-arginine in mammalian tissues is known as nitric oxide synthase (NOS) (EC.1.14.13.39). In the present study, the role of NO in the regulation of exocrine secretion was investigated in rat pancreatic acinar cells. Treatment of rat pancreatic acinar cells with cholecystokinin-octapeptide (CCK-OP) resulted in an increase in the arginine conversion to citrulline, the amount of $NO_X$, the release of amylase, and the level of CGMP. Especially, CCK-OP-stimulated increase of arginine to citrulline transformation, the amount of $NO_X$, and CGMP level were completely counteracted by the inhibitor of NOS, NG-monomethyl-L-arginine (MMA), by contrast, that of amylase release was partially reduced. Furthermore, MMA-induced decrease of NOS activity and amylase release showed dose-dependent pattern. The data on the time course of CCK-OP-induced citrulline formation and CGMP rise indicate that NOS and guanylate cyclase were activated by treatment of CCK-OP. However, the mechanism of agonist-stimulated guanylate cyclase activation in acinar cells remains unknown. Therefore, activation of NOS is one of the early events in receptor-mediated cascade of reactions in pancreatic acinar cells and NO, not completely, but partially mediate pancreatic enzyme exocrine secretion.
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