• Journal of Applied Biological Chemistry
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• v.61 no.1
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• pp.83-91
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• 2018

Advances in bacterial and fungal genome mining uncover a plethora of cryptic secondary metabolite biosynthetic gene clusters. Guided by the genome information, targeted transcriptional derepression could be employed to determine the product of a cryptic gene cluster and to explore its biological role. Monascus spp. are food grade filamentous fungi popular in eastern Asia and several genome data belong to them are now available. We achieved transcription activation of a cryptic fungal polyketide synthase-nonribosomal peptide synthase gene Mpfus1 in Monascus purpureus ${\Delta}MpPKS5$ by inserting Aspergillus gpdA promoter at the upstream of Mpfus1 through double crossover gene replacement. The gene cluster with Mpfus1 show a high similarity to those for the biosynthesis of conjugated polyene derivatives with 2-pyrrolidone ring and the mycotoxin fusarin is the representative member of this group. The ${\Delta}MpPKS5$ is incapable of producing azaphilone pigment, providing an excellent background to identify chromogenic and UV-absorbing compounds. Activation of Mpfus1 resulted in a yellow hue on mycelia and its methanol extract exhibit a maximum absorption at 365 nm. HPLC analysis of the organic extracts indicated the presence of a variety of yellow compounds in the extract. This implies that the product of MpFus1 is metabolically or chemically unstable. LC-MS analysis guided us to predict the MpFus1 product and to propose that the Mpfus1-containing gene cluster encode the biosynthesis of a desmethyl analogue of fusarin. This study showcases the genome mining in Monascus and the possibility to unveil new biological activities embedded in it.

• The Korean Journal of Physiology
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• v.24 no.1
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• pp.39-49
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• 1990

It has been reported that bombesin induces contraction of the smooth muscle of the gastrointestinal tract. Thus, the present investigation was undertaken to see an influence of bombesin on electrical activity of the gastric smooth muscle, since electrical activity is associated with contractile activity in the smooth muscle of the stomach. Smooth muscle strips $(5\;{\times}\;1.5\;cm)$ that included the corpus and antrum were prepared from the ventral and dorsal portion of the feline stomach along the greater curvature. Circular muscle strips $(1\;{\times}\;0.3\;cm)$ of the corpus were also obtained. Electrical activity of the corpus and antrum of the muscle strip was monophasically recorded by using Ag-AgCl capillary electrodes placed on the circular muscle layer. Contractile activity of the circular muscle strip was also recorded. The recordings were performed in Krebs-Ringer solution that was continuously aerated with $O_{2}$ containing 5% $Co_{2}$, and kept at $36^{\circ}C$. Dose-related responses of electrical activity and contractility to bombesin was studied after frequency of slow waves and contraction of each strip reached to a steady state. An action of $D-leu^{13}-{\psi}\;(CH_{2}NH)-D-leu^{14}-bombesin,\;D-pro^{2}-D-trp^{7,9}-substance\;P$, tetrodotoxin, hexamethonium, atropine, phentolamine or propranolol on the effect of bombesin was also observed. 1) Bombesin increased frequency of slow waves and contractions dose-dependently at concentrations from $10^{-9}\;M\;to\;3\;{\times}\;10^{-8}\;M$. 2) The bombesin analogue at a concentration of $3\;{\times}\;10^{-7}\;M$ antagonized the effect of bombesin on frequency of slow waves. 3) The effect of bombesin on frequency of slow waves was inhibited by tetrodotoxin $(10^{-6}\;M)$ and hexamethonium $(10^{-3}\;M)$ but unaffected by atropine $(10^{-6}\;M)$, phentolamine $(10^{-5}\;M)$ and propranolol $(10^{-5}\;M)$. 4) The effect of bombesin on frequency of slow waves was blocked by the substance P analogue at a concentration of $10^{-5}\;M$. 5) Substance P at a concentration of $10^{-5}\;M$ failed to change frequency of slow waves. It is concluded from the above results that bombesin increases the frequency of slow waves as well as contractions of the smooth muscle strip from the feline stomach, and the effect of bombesin might be mediated by non-cholinergic or non-adrenergic mechanism at neuromuscular junction. However, enteric nerves that have substance P as a neurotransmitter do not appear to participate in the action of bombesin on frequency of slow waves.

• Korean Chemical Engineering Research
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• v.52 no.4
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• pp.503-515
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• 2014

A lot of existing wastewater treatment plants (WWTPs) are rebuilt or retrofitted for advanced wastewater treatment processes to cope with reinforced effluent criteria of nitrogen and phosphorous. Moreover, how to treat the wasted sludge from WWTPs has been also issued since the discharge of the wasted sludge into ocean is impossible from 2011 due to the London Convention 97 protocol. These trend changes of WWTPs get a motivation to assess environmental and economic impacts from the construction stage to the waste stage in WWTPs. Therefore, this study focuses on evaluation of environmental and economic impacts of the advanced wastewater treatment processes and waste sludge treatment process by using life cycle assessment. Four advanced wastewater treatment processes of Anaerobic/Anoxic/Oxic ($A_2O$), 5 stages-Bamard Denitrification Phosphate (Bardenpho), Virginia Initiative Plant (VIP), and Modified University of Cape Town (MUCT) are chosen to compare the conventional activated sludge (CAS) and three waste sludge treatment methods of land fill, incineration, and composting are used. To evaluate environmental and economic impacts of each advanced wastewater treatment processes, life cycle assessment (LCA) and life cycle cost (LCC) are conducted based on International organization for standardization (ISO) guidelines. The results clearly represent that the $A_2O$ process with composting shows 52% reduction in the environmental impact than the CAS process with landfill. On the other hand, the MUCT process with composting is able to save 62% of the life cycle cost comparing with the CAS process with landfill. This result suggested the qualitative and quantitative criteria for evaluating eco-environmental and economic technologies of advanced treatment processes and also sludge treatment method, where their main influence factors on environmental and economic impacts are analyzed, respectively. The proposed method could be useful for selecting the most efficient and eco-friendly wastewater treatment process and sludge treatment method when retrofitting the existing WWTPs to advanced treatments.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.26-27
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• 2018

With entry into force of the Nagoya Protocol to promote the fair sharing of the benefits of accessing and utilizing genetic resources, much support has been given to research on the development of biomaterials and products using domestic natural resources. Conservation and resource-saving of native species became very important through Nagoya Protocol enactment. The trend of cosmetic industry has been shifing from use synthetic chemicals to natural biomaterials, due to the safety regulations on new materials, ban on animal experiments, and expansion of cosmeceuticals range. In addition, functional cosmetic range has been expanded from whitening, wrinkle improvement, and ultraviolet shielding, to hair loss, hair loss alleviation, acne relaxation, and moisturizing of atopic skin, thus causing the activation of research about field of efficacy evaluation on natural biomaterials and commercialization. Chungbuk province is fostering the bio industry as a key industry for regional economic growth. For this purpose, Osong Biotechnology Complex/Ochang Science Industrial Complex in middle area, Jecheon biovalley in northern region, and Chungju Enterprise city have been established, thus playing a pivotal role in Bio innovative cluster in Korea. In particular, it was established the osong cosmetics clinical research support center to develop the cosmetics industry in chungbuk, thereby supporting clinical trials, efficacy evaluations, overseas certification, and overseas market entry in order to advance into the global market. In addition, oriental plants such as astragalus propinquus, schisandra chinensis, eucommia, alpiniae oxyphyllae fructus and biancaea sappan are being actively studied as global cosmetic ingredients through the promotion of various national research and development projects using natural materials in chungbuk province. The chungbuk natural product industry is expected to grow further throughout cosmetics industry development in the future, as companies and research institutes are actively promoting the secure index of effective material in natural products and effective material commercialization.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.44 no.3
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• pp.249-258
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• 2018

Airborne pollution causes oxidative damage, inflammation, and premature aging of skin. Resveratrol is a polyphenol compound that has various biological activities such as antioxidant, anti-inflammation, and anti-melanogenic activities but it is unstable to heat and light. Resveratryl triacetate (RTA) is a new cosmetic ingredient that is more stable than resveratrol and its skin safety and whitening efficacy have been reported previously. The purpose of this study was to examine the effects of resveratrol and resveratryl triacetate (RTA) on the inflammatory responses of human epidermal keratinocytes (HEKs) exposed to airborne particulate matters with a diameter of < $10{\mu}m$ (PM10). Cultured HEKs were exposed to PM10 in the absence or presence of resveratrol and RTA. Assays were undertaken to determine cell viability, the production of reactive oxygen species (ROS), and the expression of inflammatory cytokines. PM10 treatment decreased cell viability, and increased the expression of pro-inflammatory cytokines such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), $interleukin-1{\beta}$ ($IL-1{\beta}$), interleukin-6 (IL-6), and interleukin-8 (IL-8). Resveratrol and RTA reduced cell death and ROS production induced by PM10. PM10-induced mRNA expression of the inflammatory cytokines was either attenuated (IL-6), or enhanced ($IL-1{\beta}$), or unaffected ($TNF-{\alpha}$ and IL-8) by resveratrol and RTA. PM10-induced IL-6 protein expression was attenuated by resveratrol and RTA. This study suggests that resveratrol and RTA have activities regulating cell damage and inflammatory responses of the skin exposed to airborne particulate matters.

• Development and Reproduction
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• v.10 no.2
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• pp.105-113
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• 2006

Reactive oxygen species(ROS) generated in cellular metabolism have an effect on cell maturation and development. In human reproductive tract, oxidative injury by ROS may induce female infertility. Also, oxidative injury may be responsible for developmental retardation and arrest of mammalian preimplantation embryos. Activating transcription factor 4(ATF4) is a member of the cyclic-AMP response element-binding(CREB) familiy of basic region- leucine zipper(bZip). ATF4 is known to regulate stress response to protect cell from various stress factors and inducer of apoptisis. The purpose of this study was to investigate whether ATF4 is involved in the defensive mechanism in oxidative stress condition during the development of mouse preimplantation embryos. To verify the expression of ATF4 in oxidative stress condition, 2-cell stage embryos were cultured in HTF media containing 0.1mM, 0.5mM or 1mM hydrogen peroxide($H_2O_2$) for 1hr(2-cell), 8hr(4-cell), 17hr(8-cell), 24hr(morula), 48hr(early blastocyst) or 64hr(late blastocyst). The developmental rate decreased in the 0.1mM $H_2O_2$ treated group compared with control group. In embryos treated with 0.5mM and 1mM $H_2O_2$ showed 2-cell block. As a results of the semi-quantitative RT-PCR analysis of SOD1, ATF4 and Bax gene expression, SOD1, ATF4 and Bax genes were increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. In 2-cell embryos, expression of SOD1, ATF4 and Bax genes were notably increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. Immunofluorescence analysis showed that ATF4 protein was localized at the cytoplasm of preimplantation embryos. The increase in ATF4 immunoreactivety was observed in the 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. It suggests that oxidative stress by $H_2O_2$ induces expression of ATF4 and may be involved in protection mechanism in preimplantation embryos from oxidative injury.

• Korean Journal of Environmental Biology
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• v.30 no.2
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• pp.98-106
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• 2012

The chlorophyll fluorescence and photosynthetic $CO_2$ fixation capacity of leaves from three major crop trees found on Jeju Island, Camellia sinensis L., Camellia japonica L., and Citrus unshiu M., were analyzed. The photosynthetic $CO_2$ fixation rate of C. sinensis was similar to that of C. unshiu, and much higher than that of C. japonica which belongs to the same genus. Stomatal conductance in the three species was high at dawn and low during daytime. The intercellular $CO_2$ concentration of the three species was also high at dawn and decreased at midday. The transpiration rate showed an opposite trend from the intercellular $CO_2$ concentration. The photochemical efficiencies of PSII (Fv/Fm) in C. sinensis were slightly lower at midday compared to the level at dawn and/or dusk. The decline in Fv/Fm of C. sinensis at midday was much smaller than that of C. japonica. These results indicate that C. sinensis is better acclimated to high levels of radiation under natural conditions in late summer, although its PSII reaction center was inhibited by strong radiation. Of the chlorophyll fluorescence parameters in the species, the RC/CS decreased significantly while the ABS/RC, TRo/RC, ETo/RC, and DIo/RC increased significantly at midday in late summer. However, C. unshiu did not show significant changes in these values depending on the time of day. Among the three species, the daily $CO_2$ fixation rate in C. sinensis ($320.1mmol\;m^{-2}d^{-1}$) was the highest, followed by that of C. unshiu ($292.5mmol\;m^{-2}d^{-1}$) and C. japonica ($244.8mmol\;m^{-2}d^{-1}$). Thus, C. sinensis may be a valuable crop tree in terms of the uptake of $CO_2$ under natural field conditions.

• Journal of Life Science
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• v.19 no.1
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• pp.123-128
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• 2009

Production of highly valuable immunotherapeutic proteins such as monoclonal antibodies and vaccines using plant biotechnology and genetic engineering has been studied as a popular research field. Plant expression system for mass production of such useful recombinant therapeutic proteins has several advantages over other existing expression systems with economical and safety issues. Immunotherapy of multiple monoclonal antibodies, which can recognize multiple targeting including specific proteins and their glycans highly expressed on the surface of cancer cells, can be an efficient treatment compared to a single targeting immunotherapy using a single antibody. In this study, we have established plant production system to express two different targeting monoclonal antibodies in a single transgenic plant through crossing fertilization between two different transgenic plants expressing anti-colorectal cancer mAbCO17-1A and anti-breast cancer mAbBR55, respectively. The F1 seedlings were obtained cross fertilization between the two transgenic parental plants. The presence, transcription, and protein expression of heavy chain (HC) and light chain (LC) genes of both mAbs in the seedlings were investigated by PCR, RT-PCR, and immunoblot analyses, respectively. Among all the seedlings, some seedlings did not carry or transcribe the HC and LC genes of both mAbs. Thus, the seedlings with presence and transcription of HC and LC genes of both mAbs were selected, and the selected seedlings were confirmed to have relatively stronger density of HC and LC protein bands compared to the transgenic plant expressing only each mAb. These results indicate that the F1 seedling plant with carrying both mAb genes was established. Taken together, plant crossing fertilization can be applied to generate an efficient production system expressing multiple monoclonal antibodies for immunotherapy in a single plant.

• Journal of Life Science
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• v.24 no.12
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• pp.1339-1344
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• 2014

Inflammatory bowel disease is an immune disorder associated with chronic mucosal inflammation and severe ulceration in the gastrointestinal tract. Antibodies against proinflammatory cytokines, including TNF${\alpha}$, are currently used as promising therapeutic agents against the disease. Stabilization of the transcript is a crucial post-transcriptional process in the expression of proinflammatory cytokines. In the present study, we assessed the expression and histological distribution of the HuR protein, an important transcript stabilizer, in tissues from experimental animals and patients with Crohn's disease. The total and cytosolic levels of the HuR protein were enhanced in the intestinal epithelia from dextran sodium sulfate (DSS)-treated mice compared to those in control tissues from normal mice. Moreover, the expression of HuR was very high only in the mucosal and glandular epithelium, and the relative localization of the protein was sequestered in the lower parts of the villus during the DSS insult. The expression of HuR was significantly higher in mucosal lesions than in normal-looking areas. Consistent with the data from the animal model, the expression of HuR was confined to the mucosal and glandular epithelium. These results suggest that HuR may contribute to the post-transcriptional regulation of proinflammatory genes during early mucosal insults. More mechanistic investigations are warranted to determine the potential use of HuR as a predictive biomarker or a promising target against IBD.

• Journal of Life Science
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• v.30 no.2
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• pp.156-161
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• 2020

Since industrialization, the production and utilization of various chemicals has contributed to improving the quality of our lives, but the subsequent discharge of massive waste is inevitable, and environmental pollution is becoming more serious every day. Exposure to chemicals as a result of environmental pollution is having a negative effect on human health and the ecosystem, and cleaning up the polluted environment that can affect our lives is a very important issue. Toxic aromatic compounds have been detected frequently in soil, groundwater, and wastewater because of the extensive use of oil products, and phenol, which is used to produce synthetic resins, textiles, and dyes, is one of the major pollutants, along with insecticides and preservatives. Phenol can cause dyspnea, headache, vomiting, mutation, and carcinogenesis. Phenol-degrading bacterium DWB-1-8 was isolated from the activated sludge of textile wastewater; this strain was identified as Comamonas testosteroni by 16S rRNA gene sequencing. The optimal culture conditions for the cell growth and degradation of phenol were 0.7% K₂HPO₄, 0.6% NaH₂PO₄, 0.1% NH₄NO₃, 0.015% MgSO₄·7H₂O, 0.001% FeSO₄·7H₂O, an initial pH of 7, and a temperature of 30℃. The strain was also able to grow by using other toxic compounds, such as benzene, toluene, or xylene (BTX), as the sole source of carbon.