• BMB Reports
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• 제34권3호
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• pp.194-200
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• 2001

Secreted arginase from Evernia prunastri thallus has been purified 616-fold from the incubation medium. Purified arginase was resolved as only one peak in a capillary electrophoresis with a pI value of 5.35. The protein contained high amounts of acidic amino acids, such as Asx and Glx, and a relatively high quantity of Ser and Gly. The molecular mass of native, purified arginase was estimated as about 26 kDa by SE-HPLC. Substrate saturated kinetic showed a typical Michaelis-Menten relationship with a K_m value of 3.3 mM L-arginine. Atranorin behaved as a mixed activator of the enzyme (apparent $K_m$ = 0.96 mM); whereas evernic and usnic acid were revealed as non competitive inhibitors (apparent $K_m$ values were 3.16 mM and 3.05 mM, respectively). Kinetics of atranorin binding indicated that saturation was reached from 0.18 ${\mu}mol$ of the total atranorin and the occurrence of multiple sites for the ligand. This agrees with a possible aggregation of several enzyme subunits during the interaction process. A value of binding sites of about 12 was obtained. The binding of evernic acid was saturated from 23 nmol of total phenol. The number of binding sites was about 5. The loss of the binding ability of evernic acid could be interpreted as a single negative cooperatively. Usnic acid behaves in a similar way to evernic acid, although the binding saturation occurs at $0.14\;{\mu}moles$ of the ligand. This binding appears to be unspecific, and has 28 usnic acid binding sites to the protein.

• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.2763-2769
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• 2015

Background: Cancer is an unnatural type of tissue growth in which the cells exhibit unrestrained division, leading to a progressive increase in the number of dividing cells. It is now the second largest cause of death in the world. The present study concerned antioxidant, anticancer and anticholinesterase activities and protocatechuic, catechin, caffeic acid, syringic acid, p-coumaric acid and o-coumaric concentrations in methanol extracts of flowers, fruits and seeds of Hypericum amblysepalum. Materials and Methods: Antioxidant properties including free radical scavenging activity and reducing power, and amounts of total phenolic compounds were evaluated using different tests. Protocatechuic, catechin, caffeic acid, syringic acid, p-coumaric acid and o-coumaric concentrations in extracts were determined by HPLC. Cytotoxic effects were determined using the MTT test with human cervix cancer (HeLa) and rat kidney epithelium cell (NRK-52E) lines. Acetyl and butyrylcholinesterase inhibitory activities were measured by by Ellman method. Results: Total phenolic content of H. amblysepalum seeds was found to be higher than in fruit and flower extracts. DPPH free radical scavenging activity of the obtained extracts gave satisfactory results versus butylated hydroxyanisole and butylated hydroxytoluene as controls. Reducing power activity was linearly proportional to the studied concentration range: $10-500{\mu}g/mL\;LC_{50}$ values for H. amblysepalum seeds were 11.7 and 2.86 respectively for HeLa and NRK-52E cell lines. Butyryl-cholinesterase inhibitory activity was $76.9{\pm}0.41$ for seed extract and higher than with other extracts. Conclusions: The present results suggested that H. amblysepalum could be a potential candidate anti-cancer drug for the treatment of human cervical cancer, and good source of natural antioxidants.

• Biomolecules & Therapeutics
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• 제25권2호
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• pp.105-111
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• 2017

It has been known that RA, one of major constituents of Perilla frutescens which has been used as a traditional folk remedy for sedation in oriental countries, shows the anxiolytic-like and sedative effects. This study was performed to know whether RA may enhance pentobarbital-induced sleep through ${\gamma}-aminobutyric$ acid $(GABA)_A-ergic$ systems in rodents. RA (0.5, 1.0 and 2.0 mg/kg, p.o.) reduced the locomotor activity in mice. RA decreased sleep latency and increased the total sleep time in pentobarbital (42 mg/kg, i.p.)-induced sleeping mice. RA also increased sleeping time and number of falling sleep mice after treatment with sub-hypnotic pentobarbital (28 mg/kg, i.p.). In electroencephalogram (EEG) recording, RA (2.0 mg/kg) not only decreased the counts of sleep/wake cycles and REM sleep, but also increased the total and NREM sleep in rats. The power density of NREM sleep showed the increase in ${\delta}-waves$ and the decrease in ${\alpha}-waves$. On the other hand, RA (0.1, 1.0 and $10{\mu}g/ml$) increased intracellular $Cl^-$ influx in the primary cultured hypothalamic cells of rats. RA (p.o.) increased the protein expression of glutamic acid decarboxylase ($GAD_{65/67}$) and $GABA_A$ receptors subunits except ${\beta}1$ subunit. In conclusion, RA augmented pentobarbital-induced sleeping behaviors through $GABA_A-ergic$ transmission. Thus, it is suggested that RA may be useful for the treatment of insomnia.

• 한국약용작물학회지
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• 제20권6호
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• pp.479-486
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• 2012

The production of adventitious roots derived from root explant of Echinacea angustifolia and its secondary metabolite content were assessed in different types and levels of auxin. The induction of adventitious roots from root explant cultured in Murashige and Skoog solid medium supplemented with 1.0 mg/L indole -3-butyric acid (IBA) attained highest as 20.87 mg fresh weight and 3.07 mg dry weight per culture but root suspension culture at the same concentration of IBA enhanced biomass production as 3.07 g fresh weight and 0.38 g per culture after 4 weeks in culture. 3.0 mg/L ${\alpha}$-naphthalene acetic acid (NAA) treatment had similar effect on root biomass production as 3.07 g fresh weight and 0.38 g per culture with liquid suspension culture, whereas adventitious roots exposed to over 3.0-5.0 mg/L IBA or 5.0 mg/L NAA were less responsive by reducing the number of adventitious roots and/or changing root morphology such as short and thick. The content of secondary metabolites such as phenolic, flavonoids and total caffeic acid in adventitious roots cultured on MS medium supplemented with 1.0 mg/L IBA were attained highest as 27.20, 9.60. 10.67 mg/g dry weight, respectively. Overall, the best production of root biomass and secondary metabolites were given by 1.0 mg/L IBA.

• 한국발생생물학회지:발생과생식
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• 제21권2호
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• pp.205-213
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• 2017

The aim of this study was to determine the effect of additional alpha-linolenic acid (ALA) supplementation during in vitro maturation (IVM) and culture (IVC) on nucleic maturation and embryo development of pigs. Cumulus-oocyte complexes (COCs) were incubated in IVM medium containing different concentration of ALA (25, 50 and $100{\mu}M$) for 44 h. After in vitro maturation, nuclear maturation of oocytes were evaluated by aceto-orcein stain. Mature oocytes with $50{\mu}M$ ALA were fertilized and cultured in IVC medium with ALA (25, 50 and $100{\mu}M$) during early-embryogenesis (48 hours after fertilization). Then, embryos were cultured with $25{\mu}M$ ALA during early embryogenesis and/or late embryogenesis (120 hours after early-embryogenesis). In results, oocyte maturation were significantly increased by $50{\mu}M$ ALA treatment groups compared with control groups (p<0.05). Treatment of $25{\mu}M$ ALA during early-embryogenesis enhanced cleavage rate of embryo compared with other groups (p<0.05), whereas formation and total cell number of blastocyst had no significant difference. Similarly, cleavage rate of embryos were increased by $25{\mu}M$ ALA supplement during early- or late-embryogenesis than ALA treatment both stage of embryogenesis (p<0.05), but did not influence to blastocyst formation. Interestingly, total cell number of blastocyst were enhanced in ALA treatment group during early-embryogenesis. These findings indicated that ALA supplement enhance the nuclear maturation of oocyte and embryo development, however, excessive ALA could negatively influence. Therefore, we suggest that ALA is used for improvement of in vitro production of mammalian embryo and further study regarding with functional mechanism of ALA is needed.

• 한국수정란이식학회지
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• 제32권4호
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• pp.275-285
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• 2017

Lysophosphatidic acid (LPA) is an important signaling molecule. Here, the effect and mechanism of LPA on the preimplantation development of porcine embryos during in vitro culture (IVC) was examined. Porcine embryos were cultured in porcine zygote medium (PZM-3) supplemented with $30{\mu}M$ LPA during different days. There was a significantly higher cleavage rate in Day 1-7 and significantly higher total cell number of blastocysts in Day 1-3 and Day 4-7. It was also found that messenger RNA (mRNA) expression level of PCNA, BCL2 and BAX in blastocysts obtained from D1-7 group were significantly higher and BCL2/BAX mRNA ratio in D1-3 group was significantly lower than control group but Day 4-7 and Day 1-7 groups were comparable with control group. Treatment with $20{\mu}M$ PLC inhibitor significantly decreased the embryo cleavage rate and blastocyst formation rate. Moreover, LPA as an activator of PLCs, enhanced the $30{\mu}M$ LPA + $20{\mu}M$ U73122 group embryo cleavage rate which similar with control group. In conclusion, the results suggest that treatment with LPA during IVC improves the porcine early embryo cleavage by activation of PLC signaling pathway and regulate the mRNA expression that contribute to total cell number of blastocysts during blastocyst formation.

• 한국연초학회지
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• 제10권2호
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• pp.99-107
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• 1988

Pot experiment was conducted to find out the effect of lime application on yield and chemical composition of burley tobacco in 1986, Lime increased exchangeable calcium and pH of soil, but decreased Al, Fe and Mn concentrations. Yield was increased by lime application, however lime could not be caused to yield increasing in the soil with high calcium contents. Cored leaves of limed Plot contained higher Mg. K, total nitrogen, NO3-nitrogen, water soluble and insoluble ash, alkalinity number of water soluble and insoluble ash, citric and malic acid, but lower Fe, Mn, protein-nitrogen, NH3-nitrogen, nicotine petroleum ether extract, palmitic and linolenic acid concentrations than those of unlined plot. The linoleic acid and volatile neutral constituents of cured leaves were not affected by liming. Lime increased yield, however it did not affect leaf duality in respect to chemical characteristics, suggesting that liming should be necessary for tobacco cultivation.

• Korean Journal of Hydrosciences
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• 제2권
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• pp.115-126
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• 1991

The anaerobic digester with sludge from sewage treatment plant was operated in the laboratory for two year to investigate the enumeration and activity of methanogenic microorganisms. In this experimental study, the effects of HRT on the degradation characteristics of organic materials and on the number of methanogenic bacteria produced were investigated. By making the media with the repeated wxperiment, the number and activity of methanogenic bacteria were measured. The increase of the removal rate of organic acid in the digester was oberved at HRT of 2 days. The total number of methane forming bacteria estimated by the MPN method showed 2.3 $\times$ $ at HRT of 3 days, 7$\times$$ of 5 days and 7.9$\times$$ $/ml of 10 days. The optimum incubation time for measuring the number of methanogenic bacteria was found as more than four weeks. The PMA revealed 161ml CH$/l day at HRT of 10 days and the PUA 290mg COD/l day. At the incubation time 4.3 days, the maximum value of CH$ *59.1%) was found. At this time, $ was found as 15.3% and $ 25.6%.

• 유기물자원화
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• 제25권1호
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• pp.67-78
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• 2017

본 연구의 목적은 휴믹산을 농도별로 유기농 고추 유묘기와 생육기에 처리하였을 때, 고추의 발아, 생육, 수량에 미치는 영향을 검정하고자 하였다. 0.05%와 0.1% 휴믹산을 상토에 처리하였더니 무처리보다 고추 발아율이 높았으나 0.4%와 1.0% 휴믹산의 발아율은 각각 90.0%와 86.7%로 96.7%의 무처리보다 낮았다. 고추유묘 정식 30일 후, 휴믹산을 0.05%나 1.0%는 무처리에 비해 생육이 감소하였으나 0.2% 휴믹산을 처리한 고추의 평균 초장(97.6 cm), 엽수(84.7개) 및 생체중($128.1g\;plant^{-1}$)이 유의적으로 증가하였다. 휴믹산 토양관주 처리 60일 후, 0.2% 휴믹산 처리구의 고추 초장이 유의적으로 가장 길었다. 0.2%, 0.1%, 0.05% 휴믹산 처리구의 평균 청과수는 처리 간에 유의적인 차이가 없었으나 0.4%와 1.0% 휴믹산 처리구의 평균 청과수가 각각 35.2개와 29.1개로 가장 많았다. 그러나 풋고추의 생체중은 0.2% 휴믹산 처리구가 무처리에 비해 평균 $111.5g\;plant^{-1}$ 더 무거운 것으로 나타났다. 휴믹산 농도별 토양관주 60일 후, 1.0% 휴믹산 처리구를 제외하고 무처리에 비해 고추의 총무게($5.8kg\;plant^{-1}$)와 평균 무게($1.4kg\;plant^{-1}$)가 높은 것으로 나타났으며 0.2%와 0.1% 휴믹산 처리구의 고추 총무게가 각각 $9.3kg\;plant^{-1}$과 $8.6kg\;plant^{-1}$로 다른 처리농도에 비해 많았다. 휴믹산 토양관주 처리에 따른 토양의 세균, pH 및 EC에 미치는 영향을 조사하였더니, 0.2% 휴믹산에서 토양의 세균밀도가 무처리구에 비해 3.5배 더 높게 나타났다. 휴믹산의 농도가 0.05%에서 1.0%로 증가함에 따라 고추재배 토양의 pH와 EC도 증가하는 것으로 나타났다.

• 한국식품조리과학회지
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• 제20권5호
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• pp.511-519
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• 2004

The optional ingredient, Maesil juice, was adopted to improve the quality of Yulmoo Mul-Kimchi during fermentation. The final weight of the Maesil juice as a percentage of the water content in the Yulmoo Mul-Kimchi was adjusted to 0, 1, 3, 5 and 7%. The physicochemical, microbiological and sensory characteristics were determined during fermentation at 10$^{\circ}C$ over a 30 days period. After fermentation, the additions of 3 and 5% Maesil juice gave the highest pH values and lowest total acidities. With regard to the reducing sugars, the 3 and 5% treatments gave the highest contents. The number of the total cell count and lactic acid bacteria increased to their maxima during fermentation, but began to decrease during the latter stages. Here, the additions of 3 and 5% Maesil juice showed distinctive lower and higher numbers of total cells and lactic acid bacteria, respectively, during the latter stages of fermentation. With regard to the sensory evaluation on the addition of Maesil juice, the results obtained with 3 and 5% additions to the fermented Yulmoo Mul-Kimchi were favored for color, smell, sour and carbonated tastes and overall acceptability of the products. Therefore, the optimum levels of Maesil juice addition to Yulmoo Mul-Kimchi were estimated to be between 3 and 5%.