• Title/Summary/Keyword: L&R model

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Enhancement of Excretory Production of an Exoglucanase from Escherichia coli with Phage Shock Protein A (PspA) Overexpression

  • Wang, Y.Y.;Fu, Z.B.;Ng, K.L.;Lam, C.C.;Chan, A.K.N.;Sze, K.F.;Wong, W.K.R.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.637-645
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    • 2011
  • Production of recombinant proteins by excretory expression has many advantages over intracellular expression in Escherichia coli. Hyperexpression of a secretory exoglucanase, Exg, of Cellulomonas fimi was previously shown to saturate the SecYEG pathway and result in dramatic cell death of E. coli. In this study, we demonstrated that overexpression of the PspA in the JM101(pM1VegGcexL-pspA) strain enhanced excretion of Exg to 1.65 U/ml using shake-flask cultivation, which was 80% higher than the highest yield previously obtained from the optimized JM101(pM1VegGcexL) strain. A much higher excreted Exg activity of 4.5 U/ml was further achieved with high cell density cultivation using rich media. Furthermore, we showed that the PspA overexpression strain enjoyed an elevated critical value (CV), which was defined as the largest quotient between the intracellular unprocessed precursor and its secreted mature counterpart that was still tolerable by the host cells prior to the onset of cell death, improving from the previously determined CV of 20/80 to the currently achieved CV of 45/55 for Exg. The results suggested that the PspA overexpression strain might tolerate a higher level of precursor Exg making use of the SecYEG pathway for secretion. The reduced lethal effect might be attributable to the overexpressed PspA, which was postulated to be able to reduce membrane depolarization and damage. Our findings introduce a novel strategy of the combined application of metabolic engineering and construct optimization to the attainment of the best possible E. coli producers for secretory/excretory production of recombinant proteins, using Exg as the model protein.

Reduction of Allergenicity of Wheat Flour by Enzyme Hydrolysis (효소 분해에 의한 밀가루의 항원성 저감화)

  • Park, Ju-Yeon;Ahn, Jeung-Yeub;Hong, Hee-Ok;Hahn, Young-Sook
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.152-157
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    • 2004
  • Gluten was extracted from domestic wheat flour using UTH buffer (4 M urea in 0.1 M Tris-HCl, pH 8.6) and validated by SDS-PAGE analysis for production of wheat flour products with reduced gluten content.. Anti-gluten polyclonal antibody was made by administering extracted gluten fraction on animal model. Anti-gluten serum titer of extracted gluten fraction was evaluated by ELISA, and that of antibody titer according to administration period. Anti-gluten sera were used for ELISA and immunoblot analysis before and after hydrolysis of gluten fraction at optimal pH and temperature condition for each protease. Gluten fraction separated by SDS-PAGE showed several bands covering 75 to 10 kDa, in which anti-gluten sera were 25, 34, and 45 kDa. Enzyme hydrolysis of gluten fraction revealed protein band sizes to be lower than 15 kDa. Content of pretense from bovine pancreas (b.p. protease) for gluten hydrolysis was estimated as 1 mg in 10 mL gluten fraction extracted for 4 hr.

Effects of Replacing Lucerne (Medicago sativa L.) Hay with Fresh Citrus Pulp on Ruminal Fermentation and Ewe Performance

  • Sparkes, J.L.;Chaves, A.V.;Fung, Y.T.E.;van Ekris, I.;Bush, R.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.2
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    • pp.197-204
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    • 2010
  • Two studies were conducted to determine the effects of replacing 30% (% in diet DM) of lucerne (Medicago sativa L.) hay with citrus pulp in Merino ewe diets: i) an in vitro study which measured ruminal fermentation; and ii) an in vivo study in which twelve Merino ewes pre- and post-lambing were fed experimental diets in a cross-over design over 120 days to evaluate effects on ewe performance (i.e. DM intake, average daily gain (ADG) and wool growth). In both the in vitro and in vivo studies, the control treatment consisted of lucerne (91.3% in diet DM), lupins (8.3% in diet DM) and phosphate (0.42% in diet DM), while the citrus pulp treatment consisted of lucerne (57.7% in diet DM), lupins (9.5% in diet DM), phosphate (0.48% in diet DM) and fresh citrus pulp (32.3% in diet DM). Data were analysed using the mixed model procedure of SAS. In the in vitro study, gas production, total volatile fatty acid (VFA) yield, proportion of propionic acid to total VFA and in vitro dry matter digestibility (IVDMD) were higher (p<0.02) in the citrus pulp treatment compared to the control treatment. In contrast, in vitro ammonia production, pH and the acetate to propionate ratio were lower (p<0.03) for the citrus pulp treatment compared to the control treatment. In the in vivo study, DM intake of ewes fed the citrus pulp diet was lower than their control ewe counterparts throughout both the pre- and post-lambing periods (928.9 vs. 1,115.0 g/d pre-; 1,285.0 vs. 1,620.3 g/d post-lambing, p<0.01), however ADG was similar (p = 0.12). Wool growth parameters and lamb performance did not differ (p>0.32) between treatments. In summary, the in vitro study demonstrated that the replacement of 30% of a lucerne diet with fresh citrus pulp improved total VFA yield, increased total gas production and improved IVDMD, while decreasing the production of ammonia, acetic acid and rumen pH. In addition, the in vivo study demonstrated that the replacement of 30% of a lucerne diet with fresh citrus pulp pre- and post-lambing decreased intake but did not affect ewe performance in terms of ADG and wool growth. These findings, of course, would be of significant interest to sheep producers endeavouring to control cost of feed ingredients whilst maintaining productivity.

Use of Hydrogen Peroxide with Ozone to Simultaneously Reduce MIB and Quench Ozone Residual in Existing Water Treatment Plants Sourcing Water from the Han River (한강을 원수로 하는 오존/과산화수소 고도정수처리공정에서의 MIB제거 및 잔류오존 농도에 관한 연구)

  • McAdams, Stephen R.;Koo, Bon Jin;Jang, Myung Hoon;Lee, Sung Kyoo
    • Journal of Korean Society on Water Environment
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    • v.28 no.5
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    • pp.704-716
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    • 2012
  • This paper provides a detailed account of pilot testing conducted at South Lake Tahoe (California), the Ddukdo (Seoul) water treatment plant (WTP) and the Bokjung (Seongnam) WTP between February, 2010, and February, 2012. The objectives were first, to characterize the reactions of ozone with hydrogen peroxide (Peroxone) for Han River water following sand filtration, second to determine empirical ozone and hydrogen peroxide doses to remove a taste-and-odor surrogate 2-methylisoborneol (MIB) using an advanced oxidation process (AOP) configuration and third, to determine the optimum dosing configuration to reduce residual ozone to a safe level at the exit of the process. The testing was performed in a real-time plant environment at both low- and high seasonal water temperatures. Experimental results including ozone decomposition rates were dependent on temperature and pH, consistent with data reported by other researchers. MIB in post-sand-filtration water was spiked to 40-50 ng/L, and in all cases, it was reduced to below the specified target level (7 ng/liter) and typically non-detect (ND). It was demonstrated that Peroxone could achieve both MIB removal and low effluent ozone residual at ozone+hydrogen peroxide doses less than those for ozone alone. An empirical predictive model, suitable for use by design engineers and operating personnel and for incorporation in plant control systems was developed. Due to a significant reduction in the ozone reaction/decomposition at low winter temperatures, results demonstrate the hydrogen peroxide can be "pre-conditioned" in order to increase initial reaction rates and achieve lower ozone residuals. Results also indicate the method, location and composition of hydrogen peroxide injection is critical to successful implementation of Peroxone without using excessive chemicals or degrading performance.

Mitigation effects of red Platycodon grandiflorum extract on lipopolysaccharide-induced inflammation in splenocytes isolated from mice (홍도라지 추출물이 마우스에서 분리한 비장세포에서 lipopolysaccharide로 유도된 염증에 미치는 영향)

  • Park, Eun-Jung;Lee, You-Suk;Jeong, Hyun Cheol;Lee, Sung-Hyen;Lee, Hae-Jeung
    • Journal of Nutrition and Health
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    • v.52 no.3
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    • pp.243-249
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    • 2019
  • Purpose: Platycodon grandiflorum (PG) is known to have effective antimicrobial and anticancer activity. The main bioactive components of PG are saponins, and these could contribute to anti-inflammatory activity. However, little is known about the anti-inflammatory effect of PG. In this study, we aim to assess the anti-inflammatory response to Red PG Extract (RPGE) in splenocytes under ex vivo conditions. Methods: The cell viability of isolated splenocytes taken from mice was analyzed by performing a Cell Counting Kit-8 assay. The productions of nitric oxide (NO) and cytokines (specifically interleukin-6 (IL-6) and interleukin-10 (IL-10)) were measured utilizing Griess reagent and ELISA, respectively. Results: We found that co-treatment with RPGE and Lipopolysaccharide (LPS) decreased isolated splenocyte proliferation as compared with that of the LPS-stimulated control. We also observed that RPGE markedly suppressed NO synthesis and IL-6 production that was induced by LPS. There were no significant differences of IL-10 production between co-treatment with RPGE plus LPS and treatment with LPS alone. Conclusion: When taken together, our data has shown that RPGE mitigates LPS-induced inflammation in splenocytes isolated from mice. Further research is surely needed to confirm the anti-inflammation effects of RPGE in an in vivo model.

Development Model of the Foxglove Aphid, Aulacorthum solani (Kaltenbach) on Lettuce (상추에서의 싸리수염진딧물(Aulacorthum solani)의 발육과 발육모형)

  • Lee, Sang-Guei;Kim, Hyeong-Hwan;Kim, Tae-Heung;Park, Gil-Jun;Kim, Kwang-Ho;Kim, Ji-Soo
    • Korean journal of applied entomology
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    • v.47 no.4
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    • pp.359-364
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    • 2008
  • The development of Aulacorthum solani (Kaltenbach) was studied at temperatures ranging from 12.5 to $27.5^{\circ}C$ under $65{\pm}5%$ RH, and a photoperiod of 16:8 (L:D). Mortality of $1st{\sim}2nd$ nymph was higher than that of $3rd{\sim}4th$ nymph at the most temperature ranges whereas at high temperature of $27.5^{\circ}C$, more $3{\sim}4th$ nymph stage individuals died. The total developmental time ranged from 16.9 days at $12.5^{\circ}C$ to 6.6days at $22.5^{\circ}C$, suggesting that higher the temperature, faster the development. However, at higher temperature of $25^{\circ}C$ the development took 7.4 days. The lower developmental threshold temperature and effective accumulative temperatures for the total immature stage were $0.08^{\circ}C$ and 162.8 day-degreeslated development. The nonlinear shape of temperature rewas well described by the modified Sharpe and DeMichele model. When the normalized cumulative frequency distributions of developmental times for each life stage were fitted to the three-parameter Weibull function, attendance of shortened developmental times was apparent with in $1{\sim}2nd$ nymph, $3{\sim}4th$ nymph, and total nymph stages in descending order. The coefficient of determination $r^2$ ranged between 0.86 and 0.91.

Comparison of Development times of Myzus persicae (Hemiptera:Aphididae) between the Constant and Variable Temperatures and its Temperature-dependent Development Models (항온과 변온조건에서 복숭아혹진딧물의 발육비교 및 온도 발육모형)

  • Kim, Do-Ik;Choi, Duck-Soo;Ko, Suk-Ju;Kang, Beom-Ryong;Park, Chang-Gyu;Kim, Seon-Gon;Park, Jong-Dae;Kim, Sang-Soo
    • Korean journal of applied entomology
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    • v.51 no.4
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    • pp.431-438
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    • 2012
  • The developmental time of the nymphs of Myzus persicae was studied in the laboratory (six constant temperatures from 15 to $30^{\circ}C$ with 50~60% RH, and a photoperiod of 14L:10D) and in a green-pepper plastic house. Mortality of M. persicae in laboratory was high in the first(6.7~13.3%) and second instar nymphs(6.7%) at low temperatures and high in the third (17.8%) and fourth instar nymphs(17.8%) at high temperatures. Mortality was 66.7% at $33^{\circ}C$ in laboratory and $26.7^{\circ}C$ in plastic house. The total developmental time was the longest at $14.6^{\circ}C$ (14.4 days) and shortest at $26.7^{\circ}C$ (6.0 days) in plastic house. The lower threshold temperature of the total nymphal stage was $3.0^{\circ}C$ in laboratory. The thermal constant required for nymphal stage was 111.1DD. The relationship between developmental rate and temperature was fitted nonlinear model by Logan-6 which has the lowest value on Akaike information criterion (AIC) and Bayesian information criterion (BIC). The distribution of completion of each developmental stage was well described by the 3-parameter Weibull function ($r^2=0.95{\sim}0.97$). This model accurately described the predicted and observed occurrences. Thus the model is considered to be good for use in predicting the optimal spray time for Myzus persicae.

In Vivo Image and Biodistribution of $^{123}I$-15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid (BMIPP) in Liposarcoma Bearing Nude Mice (지방육종형성 동물모델에서 $^{123}I$-15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid (BMIPP)의 생체분포와 생체영상)

  • Lee, Tae-Sup;Suh, Yong-Sup;Choi, Chang-Woon;Woo, Kwang-Sun;Chung, Wee-Sup;Lim, Soo-Jung;Lim, Sang-Moo;Awh, Ok-Doo
    • The Korean Journal of Nuclear Medicine
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    • v.35 no.5
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    • pp.324-333
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    • 2001
  • Purpose: $^{123}I$-labeled fatty acids have been used in the evaluation of regional myocardial energy metabolism. This study aimed to evaluate the usefulness of $^{123}I$-BMIPP as a liposarcoma-imaging agent. Materials and Methods: We compared in vitro uptakes between liposarcoma(SW872) and glioma(9L) cell lines, and examined biodistribution and in vivo images of $^{123}I$-BMIPP in liposarcoma-bearing nude mice. Cold-BMIPP was labeled with $^{123}I\;using\;Cu^{2+}$ as catalyst. After purification by Sep-pak, radiochemical purity was determined by TLC. We compared cellular uptake between glioma and liposarcoma after incubation of 5, 10, 15, 30, 60, 120, and 180 mins with culture medium containing $^{123}I$-BMIPP. The difference in biodistribution was determined between non-feeding (water only) group for 18 hr and feeding group in normal mice (n=6/group) at 0.5, 2, and 24 hr. In liposarcoma-hearing nude mice model, liposarcoma, SW872, ceil lines were injected subcutaneously into the felt thigh of nude mice. The biodistribution of $^{123}I$-BMIPP was evaluated at 0.5, 2, and 24 hr (n:5 / group) and in vivo Image of $^{123}I$-BMIPP was obtained with gamma camera at 2 and 24 hr in liposarcoma-hearing nude mice. Results: Radiolabeling yield and radiochemical purity were 95% and above 99%, respectively. SW872 cell line showed more increased uptake than 9L with 1.5 times at 180 mins. The clearance of $^{123}I$-BMIPP in various tissues was more delayed in the non-feeding group than in the feeding group, especially at delayed time (24 hr) in normal mice, and the major excreting organ was the gastrointestinal tract. In liposarcoma-bearing nude mice, tumor/blood ratio of $^{123}I$-BMIPP was 0.94, 0.75, and 1.38 and tumor/muscle ratio was 0.66, 1.53, and 1.11 at 0.5, 2, and 24hr, respectively. $^{123}I$-BMIPP was selectively localized in liposarcoma at 24 hr image. Conclusions: These results suggest that $^{123}I$-BMIPP can be used as a liposarcoma-imaging agent.

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Die dialektische Aporetik bei Aristoteles und ihre methodologische Funktion (아리스토텔레스에게서 아포리아의 변증적 탐구술과 그 방법론적 기능)

  • Lee, Jae-hyun
    • Journal of Korean Philosophical Society
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    • v.126
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    • pp.263-293
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    • 2013
  • Die vorliegende Abhandlung beachtet die methodologische $N{\ddot{u}}tzlichkeit$ der Dialektik $f{\ddot{u}}r$ die philosophische $Wissenschaftst{\ddot{a}}tigkeit$ des Aristoteles in einer besonderen Aufmerksamkeit auf den Begriff "${\delta}{\iota}{\alpha}{\pi}o{\rho}{\varepsilon}{\iota}{\nu}$". Bei Aristoteles bedeutet "${\delta}{\iota}{\alpha}{\pi}o{\rho}{\varepsilon}{\iota}{\nu}$" 'das durchaus $gr{\ddot{u}}ndliche$ Untersuchen einer Aporie', und eine solche Untersuchungsweise folgt der Dialektik. Deshalb beabsichtige ich zuerst, durch eine Begriffsanalyse und - interpretation von "${\delta}{\iota}{\alpha}{\pi}o{\rho}{\varepsilon}{\iota}{\nu}$" die dialektische Aporetik zu bestimmen als eine Argumentationsmethode nicht nur in einer dialogischen, sondern auch in einer nicht-dialogischen Situation. Besonders will ich einen methodologischen Grund $daf{\ddot{u}}r$ suchen, dass ein allein Forschender in der nicht-dialogischen Situation die dialektische Aporetik verwendet, indem ich einen Text von "De caelo" II 13, 294b 6~13 interpretiere. $N{\ddot{a}}mlich$ betont Aristoteles in Bezug auf den Durchgang der $Wissenschaftst{\ddot{a}}tigkeit$ eine Forderung der selbstkritischen Haltung, die der allein Forschende sich selbst seine Untersuchung in $R{\ddot{u}}cksicht$ auf die anderen Untersuchungen und auch auf die vorhersehbaren Hypothesen ${\ddot{u}}berpr{\ddot{u}}fen$ und verifizieren, um die aus seiner Untersuchung zu entdeckenden Aporien auf die dialektische Weise zu $l{\ddot{o}}sen$. Und diese Position von Aristoteles kann der wichtige Grund $daf{\ddot{u}}r$ sein, den fachsprachlichen Sinn von "${\delta}{\iota}{\alpha}{\pi}o{\rho}{\varepsilon}{\iota}{\nu}$" als auch den wissenschaftlichen Sinn der dialektischen Untersuchung der Aporie richtig verstehen und $erkl{\ddot{a}}ren$ zu $k{\ddot{o}}nnen$. Zuletzt bin ich in der Absicht, die dialektische Aporetik und ihre Funktion in drei Modeln zu formulieren und einige Beispiele $f{\ddot{u}}r$ jedes Model vorzustellen, um zu zeigen, auf welche Art und Weise Aristoteles die dialektische Aporetik $f{\ddot{u}}r$ den Lauf seiner Theoriebildung in der Tat anwendet. Durch solchen Versuch $w{\ddot{u}}nsche$ ich, dass meine Untersuchung ${\ddot{u}}ber$ die dialektische Aporetik etwas besseres $Verst{\ddot{a}}ndnis$ $f{\ddot{u}}r$ die wissenschaftliche Bedeutung der aristotelischen Dialektik anbieten $k{\ddot{o}}nnte$, und auch dass diese Studie als bedeutsame Basisdaten zu einer noch tieferen Forschung und $Verst{\ddot{a}}ndigung$ ${\ddot{u}}ber$ die philosophische Haltung und Methode des Aristoteles betrachtet werden $k{\ddot{o}}nnte$.

Biogas Production from Vietnamese Animal Manure, Plant Residues and Organic Waste: Influence of Biomass Composition on Methane Yield

  • Cu, T.T.T.;Nguyen, T.X.;Triolo, J.M.;Pedersen, L.;Le, V.D.;Le, P.D.;Sommer, S.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.2
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    • pp.280-289
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    • 2015
  • Anaerobic digestion is an efficient and renewable energy technology that can produce biogas from a variety of biomasses such as animal manure, food waste and plant residues. In developing countries this technology is widely used for the production of biogas using local biomasses, but there is little information about the value of these biomasses for energy production. This study was therefore carried out with the objective of estimating the biogas production potential of typical Vietnamese biomasses such as animal manure, slaughterhouse waste and plant residues, and developing a model that relates methane ($CH_4$) production to the chemical characteristics of the biomass. The biochemical methane potential (BMP) and biomass characteristics were measured. Results showed that piglet manure produced the highest $CH_4$ yield of 443 normal litter (NL) $CH_4kg^{-1}$ volatile solids (VS) compared to 222 from cows, 177 from sows, 172 from rabbits, 169 from goats and 153 from buffaloes. Methane production from duckweed (Spirodela polyrrhiza) was higher than from lawn grass and water spinach at 340, 220, and 110.6 NL $CH_4kg^{-1}$ VS, respectively. The BMP experiment also demonstrated that the $CH_4$ production was inhibited with chicken manure, slaughterhouse waste, cassava residue and shoe-making waste. Statistical analysis showed that lipid and lignin are the most significant predictors of BMP. The model was developed from knowledge that the BMP was related to biomass content of lipid, lignin and protein from manure and plant residues as a percentage of VS with coefficient of determination (R-square) at 0.95.This model was applied to calculate the $CH_4$ yield for a household with 17 fattening pigs in the highlands and lowlands of northern Vietnam.