• Journal of Life Science
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• v.17 no.2 s.82
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• pp.198-203
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• 2007

DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.

• Parasites, Hosts and Diseases
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• v.47 no.2
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• pp.117-124
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• 2009

Toxoplasma gondii provokes rapid and sustained nuclear translocation of the signal transducer and activator of transcription 6 (STAT6) in HeLa cells. We observed activation of STAT6 as early as 2hr after infection with T. gondii by the nuclear translocation of fluorescence expressed from exogenously transfected pDsRed2-STAT6 plasmid and by the detection of phosphotyrosine-STAT6 in Western blot. STAT6 activation occurred only by infection with live tachyzoites but not by co-culture with killed tachyzoites or soluble T. gondii extracts. STAT6 phosphorylation was inhibited by small interfering RNA of STAT6 (siSTAT6). In view of the fact that STAT6 is a central mediator of IL-4 induced gene expression, activation of STAT6 by T. gondii infection resembles that infected host cells has been stimulated by IL-4 treatment. STAT1 was affected to increase the transcription and expression by the treatment of siSTAT6. STAT6 activation was not affected by any excess SOCS's whereas that with IL-4 was inhibited by SOCS-1 and SOCS-3. T. gondii infection induced Eotaxin-3 gene expression which was reduced by $IFN-{\gamma}$. These results demonstrate that T. gondii exploits host STAT6 to take away various harmful reactions by $IFN-{\gamma}$. This shows, for the first time, IL-4-like action by T. gondii infection modulates microbicidal action by $IFN-{\gamma}$ in infected cells.

• The Korean Journal of Ecology
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• v.21 no.5_1
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• pp.465-473
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• 1998

This study was carried out to investigate the developmental process of plant community during the secondary succession and changes of soil properties in the burned areas lapsed 28 years after the forest fire in Mt.Palgong. The forest fire occurred on March, 1969 and the red pine (pinus densiflora) forest and its floor vegetation were burned down. The results are summarized as follows: the floristic composition of burned and unburned areas were composed of 49 and 48 species of vascular plants, respectively. The dominant species based on SDR4 of the burned sites were Lespedeza maximowicxii(87.75), Carex humilis (62.94), Rhododendron schippenbachii(55.78) and Miscanthus sinensis var.purpurascens (51.94). In contrast, Pinus densiflora (81.17), Quercus serrata (53.58)m Carex humilis (53.11) and Miscanthus sinenis var. purpuracens (52.42) were dominant in the unburned area. The biological spectra showed the $H-D_1-R_5-e$ type in both areas. The indices of similarity (CCs) between the two areas were 0.80. Degree of succession (DS) was 734 in the burned area and 809 in the unburned area. The species diversity (H) and evenness indices (e) in the burned and unburned areas were 2.05, 2.13 and 0.53, 0.55, respectively. Dominance index (C) in the burned and unburned areas were 0.30 and 0.32, respectively. Soil properties such as soil pH, content of organic matter, total nitrogen, total carbon, exchangeable potassium, sodium, calcium, and magnesium in burned area were comparatively higher than those of unburned area. Monthly changes of soil properties were of little significance except for some cases. These results suggest that there was relationship between trend of vegetation recovery and the changes of soil properties after the forest fire. Mixed forestation of fire-resistant species and nitrogen fixation species will be effective for reforestation after the forest fire.

• Biomolecules & Therapeutics
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• v.31 no.1
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• pp.127-138
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• 2023

Glycogen synthase kinase-3β (GSK-3β) is an important serine/threonine kinase that implicates in multiple cellular processes and links with the neurodegenerative diseases including Alzheimer's disease (AD). In this study, structure-based virtual screening was performed to search database for compounds targeting GSK-3β from Enamine's screening collection. Of the top-ranked compounds, 7 primary hits underwent a luminescent kinase assay and a cell assay using human neuroblastoma SH-SY5Y cells expressing Tau repeat domain (Tau_RD) with pro-aggregant mutation ΔK280. In the kinase assay for these 7 compounds, residual GSK-3β activities ranged from 36.1% to 90.0% were detected at the IC₅₀ of SB-216763. In the cell assay, only compounds VB-030 and VB-037 reduced Tau aggregation in SH-SY5Y cells expressing ΔK280 Tau_RD-DsRed folding reporter. In SH-SY5Y cells expressing ΔK280 Tau_RD, neither VB-030 nor VB-037 increased expression of GSK-3α Ser21 or GSK-3β Ser9. Among extracellular signal-regulated kinase (ERK), AKT serine/threonine kinase 1 (AKT), mitogen-activated protein kinase 14 (P38) and mitogenactivated protein kinase 8 (JNK) which modulate Tau phosphorylation, VB-037 attenuated active phosphorylation of P38 Thr180/ Tyr182, whereas VB-030 had no effect on the phosphorylation status of ERK, AKT, P38 or JNK. However, both VB-030 and VB-037 reduced endogenous Tau phosphorylation at Ser202, Thr231, Ser396 and Ser404 in neuronally differentiated SH-SY5Y expressing ΔK280 Tau_RD. In addition, VB-030 and VB-037 further improved neuronal survival and/or neurite length and branch in mouse hippocampal primary culture under Tau cytotoxicity. Overall, through inhibiting GSK-3β kinase activity and/or p-P38 (Thr180/Tyr182), both compounds may serve as promising candidates to reduce Tau aggregation/cytotoxicity for AD treatment.

• Korean Journal of Organic Agriculture
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• v.15 no.3
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• pp.319-330
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• 2007

This study analyzed physical and chemical characteristics of peat soil to use peat soil as the materials fur pot media and investigated seedling quality of horticultural plants in order to use peat soil as the raw materials fur pot media. The summary of the results is as follows; The chemical characteristics of peat soil, which is main ingredient of pot media are as follows; pH was 4.9, EC was less than $2.0ds{\cdot}m^{-1}$, which interferes the growth of the plant and organic ingredient was 33%. When looking into the germination characteristics of plants according to the mixture of pot media, red pepper showed 54.2% speed of germination and 97% germination rate in peat soil single treatment. Therefore the peatsoil was appropriate for the pot media for red pepper. In case of cucumbers, in the 50 : 50 treatment of main ingredient (peat soil) and auxiliary ingredients (vermiculite, peat moss and perlite) they showed 100% speed of germination and 100% germination rate. Therefore 50 : 50 treatment was appropriate fur the pot media for cucumbers. In case of chinese cabbage, peat soil, perlite and peat moss mixture (50 : 25 : 25) treatment showed the highest speed of germination (77.5%), while the germination rate was a little lower (92.15%) than comparative soil. However, it was appropriate for the pot media for chinese cabbage. In case of watermelon, germination was bad because of the influence of EC when the teat soil ingredient is over 80%. However, in the mixture of peat soil and vermiculite (50:50) treatment, they showed 91.6% speed of germination and 100% germination rate. Therefore it was appropriate for the pot media for watermelon. When looking into the growth of the plants according to the mixture of ingredients, peat soil and perlite (50:50) mixture showed excellent seedling quality for cucumbers, peat soil and perlite (50:50) mixture showed excellent seedling quality and it was proven to be appropriate for the pot media for cucumbers. In case of watermelon, peat soil, peat moss and perlite (80 : 10 : 10) mixture showed excellent seedling quality and it was proven to be appropriate for the pot media for watermelon.

• Korean Journal of Environmental Agriculture
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• v.32 no.3
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• pp.193-200
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• 2013

BACKGROUND: Plant factory system of an artificial light type using Light-Emitting Diodes (LEDs), fluorescent light, or metal halide lamp instead of sun light is an ultimated method for plant production without any pesticides regardless of seasonal changes. The plant factory is also completely isolated from outside environmental conditions such as a light, temperature, or humidity compared to conventional greenhouse. Light-environment control such as a quality or quantity in the plant factory system is essential for improving the growth and development of plant species. However, there was little report that the effects of various light qualities provided by LEDs on Ledebouriella seseloides growth under the plant factory system. METHODS AND RESULTS: Ledebouriella seseloides seedlings transplanted at urethane sponge were grown in the plant factory system of a horizontal type with LED artificial lights for 90 days. Yamazaki solution for hydroponic culture of the seedlings was regularly irrigated by the deep flow technique (DFT) system on the culture gutters. Electrical Conductivity (EC) and pH of the solution was recorded at 1.4 ds/m and 5.8 in average, respectively during the experimental period. Number of unfolded leaves, leaf length, shoot fresh and dry weight of the seedlings were three times measured in every 30 days after beginning of the experiment. Blue LEDs, red LEDs, and fluorescent lights inside the plant factory were used as light sources. Conventional fluorescent lamps were considered as a control. In all the treatment, light intensity was maintained at $100{\mu}mol/m^2/s$ on the culture bed. Fresh weight of the seedlings was 3.7 times greater in the treatment with the mixture radiation of fluorescent light and blue+red LEDs (1:3 in energy ratio; Treatment FLBR13) than in fluorescent light treatment (Treatment FL). In FLBR13 treatment, dry weight per seedling was two times greater than in FL or BR11 treatment of blue+red LEDs (1:3 in energy ratio; Treatment BR11) during the culture period. Increasing in number of unfolded leaves was also significantly affected by the FLBR13 treatment comparing with BR11 treatment. CONCLUSION(S): Hydroponic culture of Ledebouriella seseloides seedlings was successfully achieved in the plant factory system with mixture lights of blue, red LEDs and fluorescent lights. Shoot growth of the seedlings was significantly promoted by the FLBR13 with the mixture radiation of fluorescent light, blue, and red LEDs under 1:3 mixture ratio of blue and red LEDs during the experimental period compared to conventional light conditions.

• Food Science and Preservation
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• v.24 no.8
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• pp.1188-1194
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• 2017

Gochujang, a traditional Korean food, is fermented by mixing red pepper powder, various grain, meju and salt. Changes in the kind of ingredients and fermentation method could increase health functionalities. In this study, in vivo anti-colitis effects of gochujang prepared with mixed grains, bamboo salt baked 3 times and meju starters on DSS-induced colitis in C57BL/6 mice were studied. We prepared gochujang prepared with mixed grains (MG), bamboo salt, and Aspergillus oryzae (A) and Baccillus subtilis (B) mixed starters (MG-AB) and gochujang prepared with MG, bamboo salt and A, B and Lactobacillus plantarum (L) mixed starters (MG-ABL). MG-AB and MG-ABL significantly increased body weight and colon length compared to the control (p<0.05). MG-ABL showed significantly decreased interleukin-6 (IL-6) expression in serum compared to the control and MG-AB group (p<0.05). MG-ABL also regulated mRNA and protein levels of pro-apoptotic Bcl-2-associated X protein (Bax) and anti-apoptotic B-cell lymphoma-2 (Bcl-2) in the mice colon tissue (p<0.05). Therefore, MG-ABL exhibited the increased anticolitis effects by inhibiting damage of colon tissue, probably by regulating a pro-inflammatory cytokine of IL-6 and regulated apoptosis related genes. These results indicated that gochujang changed with good ingredients and starters had colitis preventive effects and might be due to active compounds in mixed grain and bamboo salt, and produced by L during the fermentation of gochujang.

• The Korean Journal of Ecology
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• v.22 no.3
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• pp.145-153
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• 1999

Authors report the changes of the community structure and secondary succession after fire in Mt. Samma (333 m). Approximately six ha of the red pine (Pinus densiflora) forest and its floor vegetation were burned and cutted down. The vegetation and soil properties were investigated in the burned and unburned sites from April to October, 1998. The dominant species based on SDR₃ in the burned site were Lespedeza cyrtobotrya (96.87), Quercus serrata (77.90), Cyperus amuricus (46.22) and Miscanthus sinensis var. purpurascens (38.33), whereas the dominant species in the unburned site were Pinus densiflora (100.00) and Q. variabilis (66.10) at the tree layer, Q. serrata (100.00) and Zanthoxylum schinifolium (29.64) at the shrub layer., and Q. serrata (76.30) and M. sinensis var. purpurascens (72.84) at the herb layer. The biological spectra based on SDR₃ were Th-D₁-R/sub 5/-e for the burned and H (M)-D₁-R/sub 5/-e type for the unburned site, respectively. The index of similarity (CCs) between the burned and unburned sites was 0.41. The degree of succession (DS) were 609 for the burned and 1168 for the unburned site, respectively. The species diversity (H) and evenness indices (e) of the burned site were lower than those of the unburned site, but the dominance indices (C) was higher in the burned site. In the analysis of soil properties, pH, the content of NO₃/sup -/-N, available phosphrous, and exchangeable cations (K/sup +/, Ca/sup 2+/, Na/sup +/, Mg/sup 2+/) in the burned site were higher than those in the unburned site, whereas the contents of organic matter, total carbon, total nitrogen and NH₄/sup +/-N were lower in the burned site. The results show that dominant species in the burned site change from Miscanthus-Lespedeza→Lespedeza→Quercus and finally to Quercus forest, and the changes of soil properties also affect the early vegetational succession after fire.

• Korean Journal of Soil Science and Fertilizer
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• v.28 no.4
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• pp.350-355
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• 1995

This study was conducted to use oyster shells, which have caused environmental problems in the coastal of Korea, as an agricultural material after processing. Physico-chemical components and neutralizing amount on the Ihyun silt loam of crushed oyster shell and slaked lime were examined. In field experiment, the properties of the soil, growth and yield of lettuce, cabbage, spinach, onion, red pepper and soybean were examined by the treatments of the shell(3.68ton/ha) or the lime (2.76ton/ha) with a randomized block design. Particle size of crushed oyster shell consisted of 73.4% of 1~60mesh and 26.6% larger then 61 mesh and contents of CaO, OM, and $P_2O_5$, etc. were 55.5%, 1.3%, and 0.29%, respectively. The requirement of the shell to neutralize the soil was 130~135% of the lime, but after 24months, it was the same. The application of the shell increased the contents of available $P_2O_5$ and $SiO_2$ exchangeable Ca in used soil. The shell tratment increased the leaf height, leaf width, etc. of the examined plants, and the yields 6~154% according to examined plants, as compared with the nonliminged, indicating that the shell possesses a great potential as an agricultural material with the same effectiveness as the lime.