A gene encoding a putative phospholipase D was isolated from Bacillus licheniformis and cloned into pGEM-T easy vector. The gene was expressed in E. coli BL21 (DE3) using a pET-21(a) vector containing His6 tag. Affinity purification of the recombinant phospholipase D with nickel-nitrilotriacetic acid (Ni-NTA) resin resulted major one-band by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The purified enzyme showed a molecular weight of 44 kDa. The optimum activity of enzyme was around pH 7.0 and the enzyme was also the most stable around this condition. The optimum temperature was about $40-45^{\circ}C$ and the enzyme still showed considerable activities at wide range of temperature. Among various detergents, Triton X-100 significantly increased the enzyme activity, resulting in 181% activity of control at 0.6 mM of the detergent. Calcium ion did not significantly affect the enzyme activity, suggesting that the enzyme might be classified into $Ca^{2+}$-independent PLD.
Although it is uncertain that the cause of changed pattern of the natural disaster related to water (i.e. flood and drought) is due to excessive carbon dioxide yielded from economic activity or the increased number of sunspots, it is apparent that there have been unusual climate change that directly affects the water resource management. Due to such a frequent unusual weather activities, there have been increased natural disaster and the most direct and major reason is considered as climate change. As we see, the climate change necessarily causes social costs. Especially, the effects on the water resource due to flood and drought take the considerable part of such costs. Therefore, this study is basic work to develop a new economic analysis technique to be used in pursuing appropriate adaptation project in field of the amount of cost damage through analysis of the effects of the climate change on the water resource. The models appeared in many reports for cost assessment of climate change were various (e.g., PAGE, DICE, AIM, IMAGE, MERGE, and etc.) and this report summarizes general characteristics of each model. To assess the effects of climate change of the water management, we defined the field of the water management on climate change. The results help post-study in field of the climate change's social-economic effect assessment, can be employed for the prioritizing process of the national fund's investment.
Advances in Internet technologies and the proliferation of mobile devices enabled consumers to approach a wide range of goods and services, while causing an adverse effect that they have hard time reaching their congenial items even if they devote much time to searching for them. Accordingly, businesses are using the recommender systems to provide tools for consumers to find the desired items more easily. Association Rule Mining (ARM) technology is advantageous to recommender systems in that ARM provides intuitive form of a rule with interestingness measures (support, confidence, and lift) describing the relationship between items. Given an item, its relevant items can be distinguished with the help of the measures that show the strength of relationship between items. Based on the strength, the most pertinent items can be chosen among other items and exposed to a given item's web page. However, the diversity of the measures may confuse which items are more recommendable. Given two rules, for example, one rule's support and confidence may not be concurrently superior to the other rule's. Such discrepancy of the measures in distinguishing one rule's superiority from other rules may cause difficulty in selecting proper items for recommendation. In addition, in an online environment where a web page or mobile screen can provide a limited number of recommendations that attract consumer interest, the prudent selection of items to be included in the list of recommendations is very important. The exposure of items of little interest may lead consumers to ignore the recommendations. Then, such consumers will possibly not pay attention to other forms of marketing activities. Therefore, the measures should be aligned with the probability of consumer's acceptance of recommendations. For this reason, this study proposes a model-based approach to combine those measures into one unified measure that can consistently determine the ranking of recommended items. A regression model was designed to describe how well the measures (independent variables; i.e., support, confidence, and lift) explain consumer's acceptance of recommendations (dependent variables, hit rate of recommended items). The model is intuitive to understand and easy to use in that the equation consists of the commonly used measures for ARM and can be used in the estimation of hit rates. The experiment using transaction data from one of the Korea's largest online shopping malls was conducted to show that the proposed model can improve the hit rates of recommendations. From the top of the list to 13th place, recommended items in the higher rakings from the proposed model show the higher hit rates than those from the competitive model's. The result shows that the proposed model's performance is superior to the competitive model's in online recommendation environment. In a web page, consumers are provided around ten recommendations with which the proposed model outperforms. Moreover, a mobile device cannot expose many items simultaneously due to its limited screen size. Therefore, the result shows that the newly devised recommendation technique is suitable for the mobile recommender systems. While this study has been conducted to cover the cross-selling in online shopping malls that handle merchandise, the proposed method can be expected to be applied in various situations under which association rules apply. For example, this model can be applied to medical diagnostic systems that predict candidate diseases from a patient's symptoms. To increase the efficiency of the model, additional variables will need to be considered for the elaboration of the model in future studies. For example, price can be a good candidate for an explanatory variable because it has a major impact on consumer purchase decisions. If the prices of recommended items are much higher than the items in which a consumer is interested, the consumer may hesitate to accept the recommendations.
Kim, Ji-Hyun;Kim, Hyun-Jin;Son, Jeong-Hwa;Chun, Ho-Nam;Yang, Jin-Oh;Choi, Sung-Jin;Paek, Nam-Soo;Choi, Gyoung-Hoon;Kim, Sung-Koo
Journal of Microbiology and Biotechnology
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v.13
no.6
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pp.919-925
/
2003
Alcohol consumption has numerous health consequences for the human body. For example, heavy drinking on a daily basis causes liver diseases, and certain products such as acetaldehyde produced from alcohol metabolism are more toxic than alcohol itself. Accordingly, the current study evaluated the role of Lactobacillus fermentum MG590 to enhance the removal of the toxic effect of alcohol in alcohol metabolism. The maximum activities of the alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) by L. fermentum MG590 were observed after 6 h of culture. The production of ADH and ALDH by L. fermentum MG590 was also confirmed by SDS-PAGE. Six hours after the addition of alcohol to a culture broth of L. fermentum MG590, the alcohol concentration decreased from 7.5 to 2.7%. From an in vitro evaluation based on hepatocytes, the viability of hepatocytes in a medium containing alcohol and the cytosol of L. fermentum MG590 was higher than that in a medium containing only alcohol. From an in vivo test using SD rats fed a 22% alcoholic drink, the blood alcohol concentration (BAC), glutamic-oxaloacetic transaminase (GOT), and glutamic-pyruvic transaminase (GPT) in the rats fed a medium containing L. fermentum MG590 were lower than those in the rats fed a medium containing only the alcohol drink. These results demonstrate that the ADH and ALDH produced by L. fermentum MG590 play an important role in detoxicating alcohol in vivo. Therefore, a fermentation broth of L. fermentum MG590 could be used as an effective alcohol detoxification drink.
The response of Spirulina (Arthrospira) platensis to high salt stress was investigated by incubating the cells in light of moderate intensity in the presence of 0.8 M NaCl. NaCl caused a decrease in photosystem II (PSII) mediated oxygen evolution activity and increase in photosystem I (PSI) activity and the amount of P700. Similarly maximal efficiency of PSII (Fv/Fm) and variable fluorescence (Fv/Fo) were also declined in salt-stressed cells. Western blot analysis reveal that the inhibition in PSII activity is due to a 40% loss of a thylakoid membrane protein, known as D1, which is located in PSII reaction center. NaCl treatment of cells also resulted in the alterations of other thylakoid membrane proteins: most prominently, a dramatic diminishment of the 47-kDa chlorophyll protein (CP) and 94-kDa protein, and accumulation of a 17-kDa protein band were observed in SDS-PAGE. The changes in 47-kDa and 94-kDa proteins lead to the decreased energy transfer from light harvesting antenna to PSII, which was accompanied by alterations in the chlorophyll fluorescence emission spectra of whole cells and isolated thylakoids. Therefore we conclude that salt stress has various effects on photosynthetic electron transport activities due to the marked alterations in the composition of thylakoid membrane proteins.
Proceedings of the Korean Society of Fisheries Technology Conference
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2000.05a
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pp.530-530
/
2000
The host defense system or immune system of all modern animals has their roots in very ancient organisms. After analyzing literature data concerning properties of invertebrates and vertebrates lectins we suggest that mechanism of mannans recognition may exist in marine invertebrates, as a universal mechanism for homeostasis maintenance and host defense, and mannan-binding lectins family of vertebrates has ancient precursor, as was shown for another S-type lectins family. We carried out the screening of mannan-binding type lectin among different species of echinoderms inhabiting in Piter the Grate Bay, the sea of Japan. As a result, the C-type lectins (SJL-32) specific for high mannose glycans was isolated from the coelomic plasma of the sea cucumbers Stichopus japonicus by ion-exchange chromatography on a DEAE-Toyopearl 650M, affinity chromatography on a mannan-Sepharose 6B and gel filtration on a Sephacryl S-200. SJL-32 is homodimer with molecular mass about 32 kDa on SDS-PAGE under non-reducing conditions. Protein part of the lectin has high conteins Asn, Glu, Ser. Hemagglutination of trypsin-treated O blood group human erythrocytes by SJL-32 was competitively inhibited by high-branched -D-mannan composed of -1,2 and -1,6 linked D-mannopyranose residues. In contrast, a variety of mono-, oligo-, and polysaccharides composed of residues of galactose and fucose showed absence or little inhibitory activities. The lectin activity strong depends on Ca2+ concentration, temperature and pH. Monospecific polyclonal antibodies were obtained to the lectin. As was shown by ELISA assay, antibodies to SJL-32 cross-reacted with human serum mannan-binding lectin. This data allows making conclusion about common antigenic determinants and structural homology of both lectins. In our opinion, SJL-32 belongs to evolutionary high conservative mannan-binding lectins (MBLs) family and takes part in the host defense against pathogenic microorganisms.
Jo, Angela M.;Maxwell, Annette E.;Choi, Sun-Hye;Bastani, Roshan
Asian Pacific Journal of Cancer Prevention
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v.13
no.6
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pp.2923-2930
/
2012
Background: Little is known about interest in faith-based health promotion programs among Asian American populations. Among the Christian denominations, the Seventh-day Adventist (SDA) church is known to place a strong doctrinal emphasis on health. Objectives: To understand appropriate ways to develop and implement health promotion programs and to conduct research among Korean American SDAs. Methods: We collaborated with the North American Division of Korean SDA Churches which sponsors annual week-long religious retreats for their church members. We developed and administered a 10-page questionnaire at their 2009 retreat in order to assess socio-demographic and church characteristics, religiosity, perceived relationship between health and religion, and interest and preferences for church-based health promotion programs. Results: Overall, 223 participants completed our survey (123 in Korean and 100 in English). The sample consisted of regular churchgoers who were involved in a variety of helping activities, and many holding leadership positions in their home churches. The vast majority was interested in receiving health information at church (80%) in the form of seminars, cooking classes and workshops (50-60%). Fewer respondents were interested in support groups (27%). Some interests and preferences differed between English and Korean language groups. Conclusion: Korean American SDA church retreat participants from a large geographic area are very interested in receiving health information and promoting health at their churches and can potentially serve as "agents of influence" in their respective communities.
In this study nitroreductase from Pseudomonas sp. HK-6 capable of degrading 2,4,6-trinitrotoluene (TNT) was characterized. Through a series of purification process including ammonium sulfate precipitation, DEAE-sepharose, and Q-sepharose, three different fractions I, II, and III having the enzyme activity of NTRs whose molecular weights were approximately 27 kDa were detected in fractions from HK-6 cells. Specific activity of the three fractions were approximately 4.85 unit/mg, 5.47 unit/mg, and 5.01 unit/mg, and concentrated to 9.0-, 10.1-, and 9.3-fold compared to crude extract, respectively. The optimal pH and temperature for the three NTR fractions were approximately 7.5 and $30^{\circ}C$, respectively. Metal ions, $Ag^{+}$ , $Cu^{ 2+}$, $Hg^{2+}$ inhibited approximately 70% of enzymes activities of all NTR, while $Fe^{2+}$ did not stimulate or inhibit the activities. Monitoring the effect of chemicals on the enzyme activity revealed that those NTR fractions lost enzyme activity in presence of $\beta$-mercaptoethanol, but were a little influenced by dithiothreitol, EDTA and NaCl. The three NTR fractions demonstrated enzyme activities for nitrobenzene and RDX as well as TNT.
To elucidate enzymatic properties of ${\alpha}-galactosidase$ (EC 3, 2, 1, 22) from germinated soybean, changes in the enzyme activities and oligosaccharide contents during germination of soybean were determined. ${\alpha}-Galactosidase$ from germinated soybean was purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. Their chemical and enzymatic properties was investigated. ${\alpha}-galactosidase$ activity of sobeam was maximized when it was germinated at $25^{\circ}C$ for 120 hour. Raffinose and stachyose in soybean were decomposed completely after 96 hours and 120 hours of germination, respectively. Soybean ${\alpha}-galactosidase$ was purified by 6.6 fold by ammonium sulfate fractionation, ion exchange chromatography on DEAE-Cellulose and Sephadex A-50, and gel filtration on Sephadex G-150. Its specific activity was 825 Units/mg protein and the yield was 2.5% of the total activity of crude extracts. The purified ${\alpha}-galactosidase$ of soybean was found to be homogeneous by polyacrylamide gel electrophoresis and by HPLC. Isoelectric point of soybean ${\alpha}-galactosidase$ was determined analytical isoelectric focusing to be pH 4.8. The soybean ${\alpha}-galactosidase$ was monomeric and its molecular weight was estimated to be 30,000 by SDS-PAGE. The optimal temperature and pH for the soybeam ${\alpha}-galactosidase$ activity were $40^{\circ}C$ and pH 6.0 and 75% of its activity was lost by heating at $60^{\circ}C$ for 10 min. The enzyme was appeared to have higher affinity to raffinose than to stachyose. The Km value of soybean enzyme was 5.3 mM for ${\rho}-nitrophenyl-{\alpha}-D-galactopyranoside$ and the activation energy on PNPG was calculated to be 13.02 Kcal per mole.
In the present study, overexpression, purification, and characterization of Aeropyrum pernix K1 chaperonin B in E. coli were investigated. The chaperonin $\beta$-subunit gene (ApCpnB, 1,665 bp ORF) from the hyperthermophilic archaeon A. pernix K1 was amplified by PCR and subcloned into vector pET21a. The constructed pET21a-ApCpnB (6.9 kb) was transformed into E. coli BL21 Codonplus (DE3). The transformant cell successfully expressed ApCpnB, and the expression of ApCpnB (61.2 kDa) was identified through analysis of the fractions by SDS-PAGE (14% gel). The recombinant ApCpnB was purified to higher than 94% by using heat-shock treatment at $90^{\circ}C$ for 20 min and fast protein liquid chromatography on a HiTrap Q column step. The purified ApCpnB showed ATPase activity and its activity was dependent on temperature. In the presence of ATP, ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}$ and $50^{\circ}$, respectively. Specifically, the activity of malate dehydrogenase (MDH) at $85^{\circ}$ was greatly stabilized by the addition of ApCpnB and ATP. Coexpression of pro-carboxypeptidase B (pro-CPB) and ApCpnB in E. coli BL21 Codonplus (DE3) had a marked effect on the yield of pro-CPB as a soluble and active form, speculating that ApCpnB facilitates the correct folding of pro-CPB. These results suggest that ApCpnB has both foldase and holdase activities and can be used as a powerful molecular machinery for the production of recombinant proteins as soluble and active forms in E. coli.
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