Kim, Jun Hyeong;Park, Chang Bum;Park, Jong Hyun;Kwon, Kang Mu;Hwang, In Hyun;Ma, Sang Yong;Oh, Suk-Heung;Kim, Dae Keun
Korean Journal of Pharmacognosy
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v.53
no.2
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pp.96-101
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2022
Caenorhabditis elegans model system was used to investigate the antioxidant activity of methanol extract of Melampyrum roseum (Scrophulariaceae). The ethyl acetate soluble fraction of the M. roseum methanol extract showed the best DPPH radical scavenging activity. The ethyl acetate fraction was measured for the activity of superoxide dismutase (SOD), catalase, and oxidative stress tolerance by using C. elegans along with reactive oxygen species (ROS) level. In addition, to confirm that the regulation of the stress response gene is responsible for the increased stress tolerance of C. elegans treated by the ethyl acetate fraction, SOD-3 expression was measured using a transgenic strain. As a result, the ethyl acetate fraction increased SOD and catalase activity, and decreased ROS accumulation in a dose-dependent manner. In addition, the ethyl acetate fraction-treated CF1553 worm showed higher SOD-3::GFP intensity than the control worm.
As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of the productivity of RNA depolymerase was performed and useful strains with regard to 5'-phosphodiesterase productivities were identified. For these useful strains optimum condition, the effect of various compounds on the activity of 5'-phosphodiesterase, and the optimum condition for enzyme reaction were discussed. The quantitative of 5'-mononucleotides produced by the action of 5'-phosphodiesterase was performed using anion-exchange column chromatography and their identified was done by paper chromatography, thinlayer chromatography, ultra violet spectrophotometry, and characteristic color reaction using carbazole and schiff's reagent. (1) Penicillium citreo-viride PO 2-11 and Streptomyces aureus SOA 4-21 from soil were identified as a potent 5'-phosphodiesterase producing strains. (2) Optimum culture conditions for Penicillium citreo-viride PO 2-11 strain isolated were found to be pH 5.0 and $30^{\circ}C$, and the optimum conditions for enzyme action of 5'-phosphodiesterase were pH 4.2 and $60^{\circ}C$. Best carbon source for the production of 5'-phosphodiesterase was found to be sucrose and ammonium nitrate for nitrogen source. Addition of 0.01% corn steep liquor or yeast extract exhibited 20% increase in the amount of 5'-phosphodiesterase production compared to the control. 5'-phosphodiesterase produced by this strain was activated by $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by EDTA, citrate, $Cu^{++},\;CO^{++}$. 5'-phosphodiesterase produced 5'-mononucleotide from RNA at a rate of 65.81%, and among the 5'-mononucleotides accumulated 5'-GMP only was found to have flavorous and the strain was also found lack of 5'-AMP deaminase. Productivity of flavorous 5'-GMP was found to be 186.7mg per gram of RNA. (3) Optimum culture canditions for the isolated Streptomyces aureus SOA 4-21 strain were pH 7.0 and $28^{\circ}C$, and the optimum conditions for the action of 5'-phosphodiesterase were pH 7.3 and $50^{\circ}C$. The best carbon source for 5'-phosphodiesterase production was found to be glucose and that of nitrogen was asparagine. Addition of 0.01% yeast extract exhibited increased productivity of 5'-phosphodiesterase by 40% compared to the non-added control. 5'-phosphodiesterase produced by this strain was activated by $Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by citrate, EDTA, $Cu^{++}$. It was also found that the strain produce 5'-AMP deaminase in addition to 5'-phosphodiesterase. For this reason although decomposition rate was 63.58% the accumulation of 5'-AMP, 5'-CMP, 5'-GMP and 5'-UMP occurred by the breakdown of RNA. In the course of these reaction 5'-AMP deaminase converted 60% of 5'-AMP thus produced into 5'-IMP and flavorous 5'-mono nucleotide production was significantly increased by this strain over the above mentioned one. Production rates were found to be 171.8mg per grain of RNA for 5'-IMP and 148.2mg per gram of RNA for 5'-GMP, respectively.
BACKGROUND: Excessive metals in the soil have become one of the most significant environmental problems. Phytoremediation has received considerable attention as a method for restoring the contaminated soils. The microbes having remarkable metal tolerance and plant growth-promoting abilities could also play a significant role in remediation of metal-contaminated soils, because bioaugmentation with such microbes could promote phytoextraction of metals. Therefore, the present study was focused on evaluating the phytoextraction of heavy metals (Co, Pb and Zn) in Helianthus annuus (sunflower) induced by bioaugmentation of a phosphate solubilizing bacterium. METHODS AND RESULTS: A phosphate solubilizing bacterium was isolated from metal-contaminated soils based on the greater halo size (>3 mm) with solid NBRIP agar medium containing 10 g glucose, 5 g $Ca_3(PO_4)_2$, 5 g $MgCl_2{\cdot}6H_2O$, 0.25 g $MgSO_4.7H_2O$, 0.2 g KCl, 0.1 g $(NH_4)_2SO_4$ in 1 L distilled water. Isolated bacterial strain was assessed for their resistance to heavy metals; $CoCl_2.6H_2O$, $2PbCO_3.Pb(OH)_2$, and $ZnCl_2$ at various concentrations ranging from $100-400{\mu}g/mL$ (Co, Pb and Zn) using the agar dilution method. A pot experiment was conducted with aqueous solutions of different heavy metals (Co, Pb and Zn) to assess the effect of bacterial strain on growth and metal uptake by Helianthus annuus (sunflower). The impact of bacterial inoculation on the mobility of metals in soil was investigated under laboratory conditions with 50 mL scaled polypropylene centrifuge tubes. The metal contents in the filtrate of plant extracts were determined using an atomic absorption spectrophotometer (Perkinelmer, Aanalyst 800, USA). CONCLUSION: Inoculation with Enterobacter ludwigii PSB 28 resulted in increased shoot and root biomass and enhanced accumulation of Co, Pb and Zn in Helianthus annuus plants. The strain was found to be capable of promoting metal translocation from the roots to the shoots of H. annuus. Therefore, Enterobacter ludwigii PSB 28 could be identified as an effective promoter of phytoextraction of Co, Pb and Zn from metal-contaminated soils.
To remove phosphate accumulated in the soil and water, Acinetobacter lwoffi PO8 possessing a high ability to accumulate phosphate was isolated from a active sludge. Bacterium was cultured in the liquid medium containing $150\;{\mu}g/mL$ of phosphate at $30^{\circ}C$ in different culture conditions to examine intracellular phosphate uptake. The initial pH in the range of $7.5{\sim}8.5$ was effective on the growth and phosphate uptake of the strain. Glycerol and arabinose used as a carbon sources showed 93 and 91% the phsphate uptake, respectively. Among the nitrogen sources, ammonium salt such as $NH_4NO_3$ and $(NH_4)_2SO_4$ was effectively utilized on the phosphate uptake compared with amino compounds. The rate of phosphate uptake of $NH_4NO_3$, and $(NH_4)_2SO_4$, was 95 and 96%, respectively The growth and Phosphate uptake ability in the strain were significantly promoted when metal ions were added in the medium; $Co^{2+}$, however, was not utilized by the strain. The capacity of phosphate uptake was enhanced to $10{\sim}20%$ when arginine, methionine, or lysine was added. Using $^{32}P$ to examine the uptake Pattern of intracellular phosphate, experiment result showed that polyphosphate was largely found in the fraction of intracellular inorganic phosphate of Acinetobacter lwoffi PO8.
To study the evolution mechanism of cracks in rocks with multiple defects, rock-like samples with multiple defects, such as strip-shaped through-going cracks and cavity groups, are used, and the crack propagation law and changes in AE (acoustic emission) and strain of cavity groups under different inclination angles are studied. According to the test results, an increase in the cavity group inclination angle can facilitate the initial damage degree of the rock and weaken the crack initiation stress; the initial crack initiation direction is approximately 90°, and the extension angle is approximately 75~90° from the strip-shaped through-going cracks; thus, the relationship between crack development and cavity group initiation strengthens. The specific performance is as follows: when the initiation angle is 30°, the cracks between the cavities in the cavity group develop relatively independently along the parallel direction of the external load; when the angle is 75°, the cracks between the cavities in the cavity group can interpenetrate, and slip can occur along the inclination of the cavity group under the action of the shear mechanism rupture. With the increase in the inclination angle of the cavity group, the AE energy fluctuation frequency at the peak stress increases, and the stress drop is obvious. The larger the cavity group inclination angle is, the more obvious the energy accumulation and the more severe the rock damage; when the cavity group angle is 30° or 75°, the peak strain of the local area below the strip-shaped through-going fracture plane is approximately three times that when the cavity group angle is 45° and 60°, indicating that cracks are easily generated in the local area monitored by the strain gauge at this angle, and the further development of the cracks weakens the strength of the rock, thereby increasing the probability of major engineering quality damage. The research results will have important reference value for hazard prevention in underground engineering projects through rock with natural and artificial defects, including tunnels and air-raid shelters.
Lee, Sang Hyeon;An, Lee-Sak;Park, Yeun Chul;Kim, Ho-Kyung
KSCE Journal of Civil and Environmental Engineering Research
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v.42
no.5
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pp.599-606
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2022
Strain gauges and the bridge weigh-in-motion (BWIM) method are the representative field measurement methods used for fatigue evaluationsof a steel bridge-in-service. For a fatigue reliability evaluation to assess fatigue damage accumulation, the effective stress range and the number of stress cycles applied as the fatigue details can be estimated based on the AASHTO Manual for Bridge Evaluations with the field measurement data of the target bridge. However, the procedure for estimating the effective stress range and the stress cycles from field measurement data has not been explicitly presented. Furthermore, studies that quantitatively compare differences in fatigue evaluation results according to the field measurement data type or processing method used are still insufficient. Here, a fatigue reliability evaluation is conducted using strain and BWIM data that are measured simultaneously. A frame model and a shell-solid model were generated to examine the effect of the accuracy of the structural analysis model when using BWIM data. Also, two methods of handling BWIM data when estimating the effective stress range and average daily cycles are defined. As a result, differences in evaluation results according to the type of field measurement data used, the accuracy of the structural analysis model, and the data handling method could be quantitatively confirmed.
It is well known that dichlorvos (DDVP), an organophosphate insecticide in common use, is so easily and rapidly hydrolyzed and excreted that it has usually little toxic effect on human body. In these days, however, it is widely used as an industrial and domestic insecticide and as an anthelmintic agent for animals, so that the accident of chemical poisoning occurs frequently. DDVP acts as a powerful inhibitor of carboxylic esterase, which can cause accumulation of acetylcholine at the synapses so paralysis of muscle and the transmission failure in cholinergic synapses dueing to desensitization of acetylcholin receptor may occure. Moreover accumulation of the acetylcholine brings about the elevation of the cyclic-AMP, which alters the cellular metabolisms of nucleic acid, carbohydrate, protein and lipid. Present study has undertaken to investigate the cardiotoxic effect of DDVP by electron microscopic study. A total of 30 Sprague-Dawley strain rats, weighing about 250gm were used as experimental animals. 2mg/kg/day of DDVP is intraperitonealy injected 3 times with intervals of every other day. On 1 day, 3 days, 5 days, 7 days and 14 days after drug administration, the animals were sacrified by cervical dislocation. Left ventricular cardiac muscles were resected and sliced into $1mm^3$. The specimens were embedded with Epon 812 and prepared by routine methods for electron microscopical observation. All preparations were stained with lead citrate and uranyl acetate and then observed with Hitachi-600 transmission electron microscope. The results were as follows: 1. In the cardiac muscle of DDVP treated rats, mitochondria with disorganized double membrane and mitochondrial crista, and vacuole formation in mitochondrial matrix were observed. But structures of mitochondria were recovered to normal in 14 days group. 2. In the cardiac muscle of DDVP treated rats, cisternae of sarcoplasmic reticulum were dilated and sacculated. But these changes were recovered to normal in 14 days group. 3. In the cardiac muscle of DDVP treated rats, glycogen particles around damaged myofibrils were decreased. But amount of glycogen particles were restored in 14 days group. 4. In the cardiac muscle of DDVP treated rats, disruption and discontinuation of myofilaments and disorganization of Z-disc were observed. But the structures of myofibrils were recovered to normal in 14 days group. It is consequently suggested that DDVP would induce the reversible degenerative changes on the ultrastructures in cardiac muscle of rat.
This study was carried out to investigate accumulation of metallothionein(MT) in rat liver and kidney by cadmium administration. After male rats of Sprague-Dawley strain weighing 60$\pm$5g were fed basal diet ad libitum for 4 weeks, two types of experiments were performed. In the first set of experiment, rats were divided into five groups. Control groups was fed basal diet without injection of cadmium. Dose groups of A, B, C and D were i.p. injected 0.625, 1.25, 2.5, 5mg Cd/Kg of body wt, once a day for two days. In the second set of experiment, rats were also divided into five groups. Control group was fed basal diet without injection of cadmium. Number groups of I, II, III and IV were i.p. injected 1, 2, 3, and 4 times every 24hrs, respectively and injection doses were 2.5mg Cd/Kg of body wt. in a day. In the first of experiment, hemoglobin contents in C, D groups were lower than control group. MT concentrations in liver and kidney were increased with increasing Cd injection doses to 2.5mg Cd/Kg of body wt. Liver - SH group values in C, D groups were higher than control group. Hematocrit values did not differ among groups. In the second of experiment, hemoglobin contents and hematocrit values were decreased. MT concentration in liver and kidney were progressively increased with increasing number of Cd injection. In both sets of experiments, liver MT concentrations were higher than kidney.
Hong, In-Soo;Kim, Jun-Seok;Lee, Min-Kyoung;Yim, Woo-Jong;Islam, Md. Rashedul;Boruah, Hari P. Deka;Chauhan, Puneet Singh;Han, Gwang-Hyun;Sa, Tong-Min
Korean Journal of Soil Science and Fertilizer
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v.42
no.4
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pp.317-322
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2009
Healthy seedling generation is the major concern in overcoming adverse effects of biotic and abiotic stresses during tender stage of development in vegetables and horticultural crops. Because of this, priority is given to research leading to the generation of healthy seedlings in crops subjected to transplanting and bedding. In this study, growth pouch experiments were conducted to determine the effect of inoculation of six different strains of Methylobacterium sp. namely, M. oryzae CBMB20, M. phyllosphaerae CBMB27, M. suomiense CBMB120, and Methylobacterium strains CBMB12, CBMB15 and CBMB17 on the seedling development of the vegetable crops cabbage, Chinese cabbage and cucumber; and horticultural crops tomato and red pepper. Crops treated with the test strains generally showed higher seedling dry matter accumulation compared to the control. Significantly higher accumulation was exhibited by CBMB12, CBMB17, and CBMB20 in cabbage, as well as for CBMB27 and CBMB120 on tomato and Chinese cabbage, respectively. Furthermore, all the strains promoted root elongation in cucumber and tomato seedlings while in Chinese cabbage and red pepper, root elongation was observed with CBMB120 and CBMB12 inoculation, respectively. Large scale nursery study is needed to develop a thorough protocol for healthy seedling development with the use of these strains.
Morphological difference of the renal glomerulus at different age groups have been studied in young (three month-old), adult (twelve month-old) and old (thirty month-old) Fisher strain 344 rats. Pieces of the tissues were taken from renal corticies prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1 M Millonig's phosphate buffer, pH 7.3), following by post-fixation with 1% osmium tetroxide (0.1 M Millonig's phosphate buffer, pH 7.3) and embedded within Araldite. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed under a JEM 100CX electron microscope. The mean thickness of glomerular basal lamina and Bowman's capsule were determined by measuring the thinnest portion of basal lamina, and by taking the average of 50 readings from electron micrographs at different ages. The numerical changes of the slit pores were compared based upon the numbers over the length of 10um of glomerular basal lamina. The results were as follow: 1. The thickness of glomerular basal lamina is increased during aging; 140.4 nm in young rats, 270.0 nm in adult ones, and 437.8 nm in old ones. 2. The thickness of basal lamina of parietal cells of Bowman's capsule is 187.5 nm in young rats, 914.0 nm in adult ones, and 2850.0 nm in old ones. 3. The numbers of the slit pores of basal lamina are reduced during aging, 30.3 slit pores/$10{\mu}m$ in adult ones, and 24.2 slit pores/$10{\mu}m$ in old ones. 4. Accumulation of dense intracytoplasmic filamentous material in the parietal cells of Bowman's capsule is increased in the vicinity of the basal lamina during aging. The proximal tubule-like epithelial cell in Bowman's capsule is observed at one glomerulus in a young rat. 5. The endothelial cells are edematous and form balloon-like structure protruding into capillary lumen in young and old rats. 6. Cytoplasm of the podocyte shows a variety of alteration during aging, such as swelling of mitochondria and of endoplasmic reticulum, and increase of microtubules, microfilaments, lysosomes and lamellated myelin structures, etc. Accumulation of dense intracytoplasmic material in the foot processes is increased in the vicinity of the basal lamina during aging. The podocytic membrane-like structures are seen in young and o]d rats. 7. The mesangial matrices and mesangial cells are increased during aging, and slight swelling of endoplasmic reticulum and Golgi cisternae in young and old rats.
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