• 한국균학회소식:학술대회논문집
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• 2016.05a
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• pp.19-20
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• 2016

Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.6
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• pp.836-842
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• 2009

The objectives of this study were to evaluate potential benefits of intercropping of corn with runner bean for a smallsized farming system, based on land equivalent ratio (LER) and silage yield and quality of corn intercropped with runner bean (Phaseolus vulgaris L.), in arid conditions of Turkey under an irrigation system. This experiment was established as a split-plot design in a randomized complete block, with three replications and carried out over two (consecutive) years in 2006 and 2007. Seven different mixtures (runner bean, B and silage corn sole crop, C, 10% B+90% C, 20% B+80% C, 30% B+70% C, 40% B+60%C, and 50% B+50%C) of silage corn-runner bean were intercropped. All of the mixtures were grown under irrigation. The corn-runner bean fields were planted in the second week of May and harvested in the first week of September in both years. Green beans were harvested three times each year and green bean yields were recorded each time. After the 3rd harvest of green bean, residues of bean and corn together were randomly harvested from a 1 $m^{2}$ area by hand using a clipper when the bean started to dry and corn was at the dough stage. Green mass yields of each plot were recorded. Silages were prepared from each plot (triplicate) in 1 L mini-silos. After 60 d ensiling, subsamples were taken from this material for determination of dry matter (DM), pH, organic acids, chemical composition, and in vitro DM digestibility of silages. The LER index was also calculated to evaluate intercrop efficiencies with respect to sole crops. Average pH, acetic, propionic and butyric acid concentrations were similar but lactic acid and ammonia-N levels were significantly different (p<0.05) among different mixtures of bean intercropped with corn. Ammonia-N levels linearly increased from 0.90% to 2.218 as the percentage of bean increased in the mixtures up to a 50:50 seeding ratio. While average CP content increased linearly from 6.47 to 12.45%, and average NDF and ADF contents decreased linearly from 56.17 to 44.88 and from 34.92 to 33.51%, respectively, (p<0.05) as the percentage of bean increased in the mixtures up to a 50:50 seeding ratio, but DM and OM contents did not differ among different mixtures of bean intercropped with corn (p>0.05). In vitro OM digestibility values differed significantly among bean-corn mixture silages (p<0.05). Fresh bean, herbage DM, IVOMD, ME yields, and LER index were significantly influenced by percentage of bean in the mixtures (p<0.01). As the percentage of bean increased in the mixtures up to a 50:50 seeding ratio, yields of fresh bean (from 0 to 24,380 kg/ha) and CP (from 1,258.0 to 1,563.0 kg/ha) and LER values (from 1.0 to 1.775) linearly increased, but yields of herbage DM (from 19,670 to 12,550 kg/ha), IVOMD (from 12,790 to 8,020 kg/ha) and ME (46,230 to 29,000 Mcal/ha) yields decreased (p<0.05). In conclusion, all of the bean-corn mixtures provided a good silage and better CP concentrations. Even though forage yields decreased, the LER index linearly increased as the percentage of bean increased in the mixture up to a 50:50 seeding ratio, which indicates a greater utilization of land. Therefore, a 50:50 seeding ratio seemed to be best for optimal utilization of land in this study and to provide greater financial stability for labor-intensive, small farmers.

• Korean Journal of Organic Agriculture
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• v.25 no.4
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• pp.901-916
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• 2017

This study was conducted to evaluate natural plant extracts for methane gas reduction in ruminants. Rumen fluid was collected from cannulated Hanwoo cow ($450{\pm}30kg$) consuming 400 g/kg concentrate and 600 g/kg timothy. The 15 ml of mixture comparing McDougall's buffer and rumen fluid in the ratio 2 to 1, was dispensed anaerobically into 50 ml serum bottles. Rumen fluid contents were collected and in vitro fermentation prepared control (timothy, 300 mg), ginseng, balloon flower, yucca plant, camellia, tea plant and ogapi extracts were added at the level of 5% against 300 mg of timothy as a substrate (v/w) and incubated for 3, 6, 9, 12, 24, 48, and 72 h. In vitro pH values range 6.55~7.41, this range include rumen titration. The dry matter digestibility was not differ between all treatments and control. Total gas emission was significantly higher (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Carbon dioxide emission was not differ all treatments on 9 h than in control and significantly higher (p<0.05) yucca plant, camellia and tea plant treatments on 12 h than control. Methane emission was not differ all treatments on 6 h than in control. The rumen microbial growth rate was significantly higher (p<0.05) in ginseng, balloon flower on 12 h and significantly higher (p<0.05) in ginseng, yucca plant, tea plant and ogapi treatments on 24 h than in control. Total VFA was significantly higher (p<0.05) in tea plant and ogapi treatments on 12 h than in control and significantly higher (p<0.05) in ginseng, balloon flower treatments on 48 h than in control. Acetic acid was significantly lower (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Propionic acid was significantly higher (p<0.05) in ginseng and balloon flower treatments on 48 h than in control. As a results, sixth natural plant extracts had no significant effect dry matter digestibility and negative on rumen fermentation, but not effect methane reduction.

• Journal of Korean Society of Environmental Engineers
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• v.37 no.12
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• pp.696-704
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• 2015

This study was carried out to evaluate the optimum operational condition of Semi-continuously Fed and Mixed Reactor (SCFMR) to treat the dairy cow manure and saw dust mixture. Step-wise increase in organic loading rates (OLRs) or decrease in hydraulic retention times (HRTs) were utilized until the biogas volume became significantly decreased at mesophilic temperature ($35^{\circ}C$). The optimum operating condition of the SCFMR fed with TS 13% dairy cow manure and saw dust mixture was found to be an HRTs of 25 days and its corresponding OLRs of $4.45kg\;VS/m^3-day$. At this condition the biogas and methane production rates were 1.44 v/v-d and 1.12 v/v-d (volume of biogas per volume of reactor per day), respectively and the TVS removal efficiency of 37% was achieved. The successful operation with such a high OLR was due to the high reactor alkalinity concentration of 14,500~15,600 mg/L as $CaCO_3$ as a result of the characteristic of the original substrate, dairy cow manure and saw dust mixture whose alkalinity was more than 8,000 mg/L as $CaCO_3$. The parameters for the reactor stability, the ratios of volatile acids and alkalinity concentrations (V/A) and the ratio of propionic acid and acetic acid concentrations (P/A) appeared to be 0.11 and 0.43, respectively, that were greatly stable in operation. Free ammonia toxicity was not experienced due to the long term acclimation by the reactor TS content ranged 7.2~10.4% during the entire operational period.

• Journal of Pharmaceutical Investigation
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• v.36 no.3
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• pp.209-215
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• 2006

Zaltoprofen, (2-(10,11-dihydro-10-oxodibenzo[b,f]thiepin-2-yl)propionic acid) is an NSAID with powerful anti-inflammatory effects as well as an analgesic action on inflammatory pain. The purpose of the present study was to evaluate the bioequivalence of two zaltoprofen tablets, $Soleton^{\circledR}$ (CJ Corp.) and SCD Zaltoprofen (Samchundang Pharmaceutical Co., Ltd.), according to the guidelines of the Korea Food and Drug Administration (KFDA). The release of zaltoprofen from the two zatoprofen formulations in vitro was tested using KP Vlll Apparatus ll method with various dissolution media. Twenty six healthy male subjects, $23.2{\pm}2.26$ years in age and$64.7{\pm}8.08$ kg in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After a single tablet containing 80 mg as zaltoprofen was orally administered, blood samples were taken at predetermined time intervals and the concentrations of zaltoprofen in serum were determined using HPLC with UV detector. The dissolution profiles of two formulations were similar in all tested dissolution media. The pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated, and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$, $C_{max}$ and untransformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug, $Soleton^{\circledR}$ were 6.33, 5.91 and 17.7% for $AUC_t$, $C_{max}$ and untransformed $T_{max}$, respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log 0.8 to log 1.25 (e.g.,log $1.01{\sim}1og\;1.11$ and log $0.928{\sim}1og\;1.18$ for $AUC_t$ and $C_{max}$, respectively). Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating SCD Zaltoprofen tablet was bioequivalent to $Soleton^{\circledR}$ tablet.

• Clean Technology
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• v.20 no.3
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• pp.232-240
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• 2014

The significance of thermoplastic polyolefin polypropylene (PP) lies in its potential to replace polyvinyl chloride (PVC), the most widely used material for automobile interiors (door trim, dash board), which discharges harmful compounds in certain conditions. Another benefit of PP (0.855 amorphous - 0.946 crystalline $g/cm^3$) is its low density compared to that of PVC ($1.1-1.45g/cm^3$), which reduces vehicle weight. Market demand for eco-friendly water-based adhesive/coating material is rising significantly as a substitute for solvent-based adhesive/coating material which emits VOC and causes harmful working conditions. Under such context, in this study, a series of eco-friendly waterborne polyurethane-urea primer (a paint product that allows finishing paint to adhere much better than if it were used alone) for hydrophobic PP were prepared from different mix of DMPA content, NCO/OH molar ratio, various wt% of silicone diol and various soft segment content, among which DMPA of 21 mole %, NCO/OH molar ratio of 1.2, modified silicone diol of 5 wt% and soft segment content of 73 wt% led to good adhesion strength. Additionally, the incorporation of optimum content of additives (0.5 wt% dispersing agent, 0.5 wt% levelling agent, 1.5 wt% antifoaming agent, 3.0 wt% matting agent) into the optimum waterborne polyurethane-urea also enabled good stability, levelling, antifoaming and non-glossy.

• Korean Journal of Environmental Agriculture
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• v.32 no.3
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• pp.214-223
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• 2013

BACKGROUND: This study focused on the development of an analytical method about dichlorprop (DCPP; 2-(2,4-dichlorophenoxy)propionic acid) which is a plant growth regulator, a synthetic auxin for agricultural commodities. DCPP prevents falling of fruits during their growth periods. However, the overdose of DCPP caused the unwanted maturing time and reduce the safe storage period. If we take fruits with exceeding maximum residue limits, it could be harmful. Therefore, this study presented the analytical method of DCPP in agricultural commodities for the nation-wide pesticide residues monitoring program of the Ministry of Food and Drug Safety. METHODS AND RESULTS: We adopted the analytical method for DCPP in agricultural commodities by gas chromatograph in cooperated with Electron Capture Detector(ECD). Sample extraction and purification by ion-associated partition method were applied, then quantitation was done by GC/ECD with DB-17, a moderate polarity column under the temperature-rising condition with nitrogen as a carrier gas and split-less mode. Standard calibration curve presented linearity with the correlation coefficient ($r^2$) > 0.9998, analysed from 0.1 to 2.0 mg/L concentration. Limit of quantitation in agricultural commodities represents 0.05 mg/kg, and average recoveries ranged from 78.8 to 102.2%. The repeatability of measurements expressed as coefficient of variation (CV %) was less than 9.5% in 0.05, 0.10, and 0.50 mg/kg. CONCLUSION(S): Our newly improved analytical method for DCPP residues in agricultural commodities was applicable to the nation-wide pesticide residues monitoring program with the acceptable level of sensitivity, repeatability and reproducibility.