Escherichia coli harboring pAC-LYCO4 and pDdxs was used for lycopene production. Three wild type strains of E. coli OW1, MG1655, and W3110 were compared with DH5${\alpha}$ used before for lycopene production. Lycopene productivity of E. coli MG1655 was similar to DH5${\alpha}$ and the highest among those wild type strain. Therefore, MG1655 strain was used for random transposon and NTG mutagenesis to increase lycopene productivity. Through transposon mutation, five transposon mutants with increased lycopene productivity were obtained. It was found that genes knocked out by transposon insertion were treB in Tn1 mutant, B2436 in Tn2 mutant, and rfaH in Tn3, 4, and 5 mutants. Lycopene productivity was the highest in Tn4 mutant among the Tn mutants, which was 6-fold and 8-fold higher in lycopene concentration and content, respectively, in comparison with those obtained with wild type strain. NTG4 mutant was acquired with NTG mutation. The highest lycopene productivity of 6 mg/L and 4 mg/g DCW was obtained from the NTG4 mutant when arabinose of 0.013 mM was added for induction of dxs, rate-limiting gene of MEP pathway. The lycopene productivity of NTG4 mutant was increased 18-fold and 12-fold in lycopene concentration and content, respectively when comparing with the wild type strain.
Melatonin has been known as an animal hormone. However, melatonin exists in diverse organisms including higher plants. The biosynthesis and physiological roles for melatonin in plants is still largely unknown, although both dicot and monocot plants have melatonin and some medicinal plants even contain large amounts of melatonin. In this study we detected melatonin in diverse Viola plants, in which melatonin had not been examined so far, by reverse phase HPLC analysis, demonstrating the wide existence of melatonin in the genus of Viola. We then fed tryptophan (Trp) and tryptamine (TAM) to the incubation medium for Viola leaf sections to test their effects on melatonin formation. Trp is also the hypothesized starting material of melatonin in plants, and TAM is the following intermediate produced by the decarboxylation of Trp. Trp feeding did not affect the contents of melatonin. In contrast, TAM feeding clearly increased the level of melatonin in Viola leaves. Because TAM is derived from Trp, we concluded that the Trp-TAM pathway exists in Viola plants as well. Ineffectiveness of Trp feeding to the change of melatonin contents supports the hypothesis that the decarboxylation step from Trp to TAM is the rate-limiting step in plant melatonin biosynthesis.
A perennial plant species of Ranunculaceae, Pulsatilla tongkangensis is a very rare species only found in limestone vegetation of South Korea. It is not only rare but also has great value as a horticultural resource, thus resulting in serious problems of overexploitation and habitat disturbance. For the conservation of the rare species and its development as a resource, the study was conducted with the following three aims: first, to investigate any new geographical distribution of the species; second, to identify the site condition and vegetation structure of its habitat; and third, to compare the ecology of P. tongkangensis in other microhabitats such as rock, slope and ridge area. A new distribution was found in limestone vegetation at Danyang-gun, Chungcheongbuk-do. It was, therefore, confirmed that the distribution of P. tongkangensis exactly coincides with the area of distribution of limestone, indicating a possibility that the species might be a calcicole. 87% of the microhabitats found in rocks. Compared to P. tongkangensis, at ridges or slopes, number of P. tongkangensis plants were few in rocks. It is possibly due to the lack of microhabitats that could settle in rocky areas. Once established on rocks, however, plants grew large enough to flower with higher number of leaves and flowers per plant, resulting in higher flowering rate on rocky areas. Cracks in/between rocks are likely to be safe places for the plants with favorable light conditions, abundant fine particles to root down and reduced competition for resources. Yet, further research is necessary in terms of determining whether the species is indeed a calcicole, whether other limiting factors work in its establishment stage, and genetic diversity exists in the population.
Kim, Eunju;Kim, Yoo-Sun;Kim, Kyung-Mi;Jung, Sangwon;Yoo, Sang-Ho;Kim, Yuri
Nutrition Research and Practice
/
v.10
no.1
/
pp.11-18
/
2016
BACKGROUND/OBJECTIVES: Type 2 diabetes (T2D) is more frequently diagnosed and is characterized by hyperglycemia and insulin resistance. $\small{D}$-xylose, a sucrase inhibitor, may be useful as a functional sugar complement to inhibit increases in blood glucose levels. The objective of this study was to investigate the anti-diabetic effects of $\small{D}$-xylose both in vitro and stretpozotocin (STZ)-nicotinamide (NA)-induced models in vivo. MATERIALS/METHODS: Wistar rats were divided into the following groups: (i) normal control; (ii) diabetic control; (iii) diabetic rats supplemented with a diet where 5% of the total sucrose content in the diet was replaced with $\small{D}$-xylose; and (iv) diabetic rats supplemented with a diet where 10% of the total sucrose content in the diet was replaced with $\small{D}$-xylose. These groups were maintained for two weeks. The effects of $\small{D}$-xylose on blood glucose levels were examined using oral glucose tolerance test, insulin secretion assays, histology of liver and pancreas tissues, and analysis of phosphoenolpyruvate carboxylase (PEPCK) expression in liver tissues of a STZ-NA-induced experimental rat model. Levels of glucose uptake and insulin secretion by differentiated C2C12 muscle cells and INS-1 pancreatic ${\beta}$-cells were analyzed. RESULTS: In vivo, $\small{D}$-xylose supplementation significantly reduced fasting serum glucose levels (P < 0.05), it slightly reduced the area under the glucose curve, and increased insulin levels compared to the diabetic controls. $\small{D}$-xylose supplementation enhanced the regeneration of pancreas tissue and improved the arrangement of hepatocytes compared to the diabetic controls. Lower levels of PEPCK were detected in the liver tissues of $\small{D}$-xylose-supplemented rats (P < 0.05). In vitro, both 2-NBDG uptake by C2C12 cells and insulin secretion by INS-1 cells were increased with $\small{D}$-xylose supplementation in a dose-dependent manner compared to treatment with glucose alone. CONCLUSIONS: In this study, $\small{D}$-xylose exerted anti-diabetic effects in vivo by regulating blood glucose levels via regeneration of damaged pancreas and liver tissues and regulation of PEPCK, a key rate-limiting enzyme in the process of gluconeogenesis. In vitro, $\small{D}$-xylose induced the uptake of glucose by muscle cells and the secretion of insulin cells by ${\beta}$-cells. These mechanistic insights will facilitate the development of highly effective strategy for T2D.
Bulletin of the Society of Naval Architects of Korea
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v.27
no.3
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pp.38-46
/
1990
A numerical solution method of the boundary integral equation for axisymmetric potential flows is presented. Those are represented by ring source and ring vorticity distribution. Strengths of ring source and ring vorticity are approximated by linear functions of a parameter $\zeta$ on a segment. The geometry of the body is represented by a cubic B-spline. Limiting integral expressions as the field point tends to the surface having ring source and ring vorticity distribution are derived upto the order of ${\zeta}ln{\zeta}$. In numerical calculations, the principal value integrals over the adjacent segments cancel each other exactly. Thus the singular part proportional to $\(\frac{1}{\zeta}\)$ can be subtracted off in the calculation of the induced velocity by singularities. And the terms proportional to $ln{\zeta}$ and ${\zeta}ln{\zeta}$ can be integrated analytically. Thus those are subtracted off in the numerical calculations and the numerical value obtained from the analytic integrations for $ln{\zeta}$ and ${\zeta}ln{\zeta}$ are added to the induced velocity. The four point Gaussian Quadrature formula was used to evaluate the higher order terms than ${\zeta}ln{\zeta}$ in the integration over the adjacent segments to the field points and the integral over the segments off the field points. The root mean square errors, $E_2$, are examined as a function of the number of nodes to determine convergence rates. The convergence rate of this method approaches 2.
In this paper, parameter characteristics such as pH effect, isotherm, kinetic and thermodynamic parameters and competitive adsorption of dyes including malachite green (MG), direct red 81 (DR 81) and thioflavin S (TS), which have different functional groups, being adsorbed onto activated carbon were investigated. Langmuir, Freundlich and Temkin isotherm models were employed to find the adsorption mechanism. Effectiveness of adsorption treatment of three dyes by activated carbon were confirmed by the Langmuir dimensionless separation factor. The mechanism was found to be a physical adsorption which can be verified through the adsorption heat calculated by Temkin equation. The adsorption kinetics followed the pseudo second order and the rate limiting step was intra-particle diffusion. The positive enthalpy and entropy changes showed an endothermic reaction and increased disorder via adsorption at the S-L interface, respectively. For each dye molecule, negative Gibbs free energy increased with the temperature, which means that the process is spontaneous. In the binary component system, it was found that the same functional groups of the dye could interfere with the mutual adsorption, and different functional groups did not significantly affect the adsorption. In the ternary component system, the adsorption for MG lowered a bit, likely to be disturbed by the other dyes meanwhile DR 81 and TS were to be positively affected by the presence of MG, thus resulting in much higher adsorption.
The Journal of The Korea Institute of Intelligent Transport Systems
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v.20
no.6
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pp.37-46
/
2021
Government agencies, such as police and local governments, strive to prevent traffic hazards and create a comfortable road environment by pormoting transportation and road facilities. To this end, roads and transportation facilities are enhanced and adjusted, and improvement projects in areas with frequent traffic accidents are carried out. Usually, improvement projects in areas with frequent traffic accidents vary by projects and region. Moreover, these projects are carried out under the supervision of a person in charge and related parties. Hence, civil complaints and subjectivity are reflected in deriving priorities for the improvement projects, limiting the efficiency of the project. To this end, a study was conducted to estimate the direction of improvement of the project target site. This study comprehensively considered road, traffic, and accident conditions of representative projects with high effectiveness in handling traffic accidents. The results of the study state that the accuracy of estimating the improvement project was around 88%. In addition, the study found that there was a strong relationship between traffic volume, accident rate, and accident severity in estimating the improvement direction.
Cholesterol inhibitory activity was investigated to develop the functional food from edible forest resources such as Allium victorialis var. platyphyllum and other 12 species. Among tested samples by enzyme-linked immunosorbant assay (ELISA), leaf extracts of A. victorialis var. platyphyllum inhibited 73.9% of the activities of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) which is the highly regulated and major rate-limiting of the cholesterol biosynthesis pathway. Moreover, those extracts inhibited 76.7% of squalene synthase which catalyzes the head-to-head condensation of two farnesyl pyrophosphate molecules to form squalene in the biosynthesis of cholesterol. In order to find out the compounds which would play a key role in inhibitory activity of cholesterol, kaempferol and quercetin were isolated from the dichloromethane soluble fraction of extracts of A. victorialis var. platyphyllum. Kampferol, quercetin and each soluble fraction was also subjected to the test of the mRNA expression of HMG-CoA reductase and squalene synthase by reverse transcriptase-polymerase chain reaction (RT-PCR) assay, respectively. By treating both enzymes with 10 ㎍/㎖ of kaempferol and quercetin for 24 hours, respectively, the mRNA expression was not observed, suggesting that both compounds inhibited the biosynthesis of cholesterol at mRNA level. In this regard, it could be inferred that cholesterol inhibitory activity of A. victorialis var. platyphyllum was derived from kaempferol and quercetin. Both compounds have already been found in many plant extracts including hardwood and softwood, but it might be first known that they have cholesterol inhibitory activity.
KSCE Journal of Civil and Environmental Engineering Research
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v.42
no.3
/
pp.409-415
/
2022
The length of the acceleration lane in the area of entrance terminals is calculated based on 13 PS/ton horsepower of a cargo truck in Korea, so it is generally overestimated compared with the capacity of most vehicles traveling on an entrance ramp. Most drivers have, therefore, an indiscreet tendency to enter the main lane in all sections of an acceleration lane, which affects the traffic flow of the main lane. Because of this tendency, measures are required to minimize the impact on traffic flow of the main lane. The operating speed, rate of entrance, and traffic volume for each vehicle were investigated at the entrance terminals of the interchanges (ICs) of Yangji IC, Suseok IC, Yongin IC, and Osan IC, and the level of improvement in traffic flow was analyzed via VISSIM simulation. From the VISSIM simulation analysis, 74.0 % of the total vehicles traveled over the specified speed from the nose point where drivers would be able to recognize the traffic condition of the main carriageway, or the point at which there is a simplification of the curve section. In addition, 88.6 % of the vehicles entered the main carriageway up to 0.8 points compared with the entire length of the acceleration lane. It was subsequently found that an improvement of average speed in the main carriageway and at the entrance ramp can be achieved from 60.1 km/h to 68.5 km/h by intentionally limiting the entrance point onto the main carriageway up to 0.265 points of the entrance ramp.
Park, S. H.;B. S. Kwon;J. R. Chun;J. W. Jahng;Lee, H. T.;K. S. Chung
Proceedings of the KSAR Conference
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2001.03a
/
pp.51-51
/
2001
Cocaine and amphetamine-regulated transcript (CART) is a satiety factor that is regulated by leptin. It was reported that the mice intracerebroventricularly injected with CART showed behavioral changes resembled with the typical behavioral alterations found in the mice carrying disorders in the brain serotonergic (5-HT) system. Hence, this study was conducted to find out the relationships between CART and 5-HT. We first examined the mRNA levels of CART after the injections of para-chlorophenylalanine (pCPA, 300 mg/kg i.p., single injection or daily for three consecutive days) in the rat brains by in situ hybridization using the mouse CART cDNA probe cloned in our laboratory. Systemic administrations of pCPA, a potent inhibitor of tryptophan hydroxylase, the rate limiting enzyme of 5-HT biosynthesis, acutely depletes the brain 5-HT transporter (5-HTT) in the dorsal raphe nucleus (DRN), which reuptakes terminal 5-HT. Results indicated that the mRNA level of CART significantly decreased in the arcuate nucleus, paraventricular nucleus, and lateral hypothalamic nucleus by three days of daily injection with pCPA with no noticeable change detected 24 hrs after the single injection. The message levels of 5-HTT in DRN decreased in both single and three days of injections. Secondly, to investigate whether CART affect to 5-HT, mouse genomic CART gene, which is consist of 3 exons and 2 introns and mouse neurofilament light (NF-L) chain promoter were cloned. Then, we constructed neuron specific expression vector, which was transfected into HeLa cell using lipid-mediated transfection system. Expression of GFP and CART linked to NF-L-chain promoter in the transfected HeLa cell were detected by using fluorescent microscope and RT-PCR. These results confirmed normal expression of DNA constructs in vitro. Then, to increase brain specific expression of CART in vivo transgenic mice carrying CART gene controlled the deleted NF-L-chain promoter were generated by the DNA microinjection into pronuclei of fertilized embryos. Transgenic mice were detected by Southern blot. Further study is necessary to examine CART expression and 5-HTT in these transgenic mice. Therefore, these results suggest that there maybe a positive molecular correlation between CART and 5-HT in responding to the stimuli.
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