Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.
Kim, Hyeong-Hwan;Cho, Myoung-Rae;Kang, Taek-Jun;Ahn, Seung-Joon;Jeon, Sung-Wook;Lee, Chan-Jung;Cheong, Jong-Chun
Journal of Mushroom
/
v.10
no.4
/
pp.184-190
/
2012
Monitorings and management experiment of adult Lycoriella ingenua (Diptera: Sciaridae) in artificial sawdust grown shiitake mushroom cultivation were carried out by using yellow sticky traps and Hypoaspis aculeifer (Acari: Laelapidae) at Hwaseong, Buyeo and Cheongwon in 2012. The larvae of L. ingenua reduced commercial value of the mushroom by direct eating or retarding the growth of mushroom by spongifying the sawdust medium. The number of L. ingenua caught by traps showed the highest peak in late september resulting in 127.5~1,025.7, 87.4~743.6 and 133.7~650.4 individuals per trap in Hwaseong, Buyeo and Cheongwon, respectively. Damage rate of fruiting bodies in shiitake mushroom by L. ingenua were 7.7~30.3% in Hwaseong, 6.7~25.3% in Buyeo and 5.3~26.0% in Cheongwon and showed the highest peak in late september. L. ingenua were effectively controlled when 30.3 individuals of H. aculeifer per $m^2$ inoculated for three time with 7 to 14 days of interval. In Hwaseong, Buyeo and Cheongwon, density of L. ingenua which were caught on the yellow sticky traps were 168.2, 126.1, 132.5 individuals, respectively. And, damage rate of fruiting bodies by L. ingenua were 5.9%, 5.5% and 5.4% respectively. Both of the L. ingenua density and damage rate of mushroom reduced more than 60% in all experimental site in contrast to the control treatment.
The present study was evaluated the antibacterial effect of the combination of $Coptidis$$rhizoma$, $Glycyrrhiza$$uralensis$ Fischet, $Schizandra$$chinensis$ and $Corni$$Fructus$(1:1:1) extracts(CGSC10). Furthermore, the effectiveness of CGSC10, sodium chlorate, and the combination of CGSC10 and sodium chlorate(CGSCS10) against $E.$$coli$ O157:H7 infection was studied using ICR female mice. During the incubation period, the dose of 5, 10, and 20% CGSC10 was inhibited the growth of $E.$$coli$ O157:H7 by 34.7, 60.2, and 76.4%, respectively. For 7 days after single challenge with $E.$$coli$ O157:H7, forty female ICR mice were divided into four experimental groups which were administered in drinking water with saline, 10% CGSC10, 15 mM sodium chlorate, and CGSCS10, respectively. On the 3rd day, the number of $E.$$coli$ O157:H7 in mouse feces was significantly decreased by administration of CGSC10, 15 mM sodium chlorate, and CGSCS10 ($p$ < 0.001). On the 7th day-after administration, CGSC10, sodium chlorate, and CGSCS10 were decreased the number of $E.$$coli$ O157:H7 by 27.1, 67.7, and 83.3%, respectively. According to the results of the present study, administration of CGSCS10 to mice can reduce the severity of $E.$$coli$ O157:H7 infection. In addition, it is suggested that CGSCS10 represents a good candidate for the treatment of enteric infections in domestic animals.
The present study was evaluated the antibacterial effect of the combination of Coptidis rhizoma, Lonicerae Flos, and Paeonia japonica (1:1:1) extracts (CLP1000). Also, the effectiveness of CLP1000, dioctahedral smectite (DHS), and the combination of CLP1000 and DHS (CLPS1000) against E. coli O157:H7 infection was studied using ICR female mice. During the incubation period, the dose of 10% and 20% CLP1000 were inhibited the growth of E. coli O157:H7 by 30% and 47%, respectively. For 7 days after single challenge with E. coli O157:H7, forty female ICR mice were divided into four experimental groups which were orally administered with saline, 10% CLP1000, 10% DHS, and 10% CLPS1000, respectively. On the 3rd day, the number of E. coli O157:H7 in mouse feces was significantly decreased by administration of CLP1000 (p < 0.05), DHS (p < 0.05) and CLPS1000 (p < 0.001). On the 7th day, CLP1000 (p < 0.05) and CLPS1000 p < 0.001) administration significantly decreased the number of E. coli O157:H7. According to the results of the present study, administration of CLPS1000 to mice can reduce the severity of E. coli O157:H7 infection. Also, it is suggested that CLPS100 represents a good candidate for the treatment of enteric infections in domestic animals.
Kim, Eun-Hee;Jang, Young-Ah;Kim, Sol-Bi;Kim, Han-Hyuk;Lee, Jin-Tae
Journal of Applied Biological Chemistry
/
v.61
no.3
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pp.297-304
/
2018
Coptis chinensis is used in oriental medicine for soothing, anti-inflammation, antimicrobial and antipyretic properties, and its main ingredient berberine is known to have strong antibacterial activity. In this study, we investigated the anti-microbial effect of hot water extract of Coptis chinensis (CW) on skin related microorganism and the airborne microbe, the antifungal effects of fungi, which are frequently detected in residential environments. CW showed antibacterial effect against Propionibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis, against the airborne microbe, which was collected in four different places. At the concentration of 100 mg/mL, the antimicrobial activity continued for 42 days, showed heat stability without change in the antimicrobial activity even after heat treatment. The MIC and MBC of CW against S. aureus was 0.03, 0.05 mg/mL, against S. epidermidis was 0.50, 0.75 mg/mL and against P. acne was 0.10, 0.15 mg/mL. As a result of measuring the MIC of four kinds of fungi with high detection frequency in the surrounding environment, Gliocladium virens was 65 mg/mL by determined as MIC which can inhibit one hundred percent of mycelial growth. The concentration 90 mg/mL was determined as MIC against Aureobasidium pullulans and 100 mg/mL against Penicilium pinophilum and Chaetomium globosum. CW was considered a safe extract that showed no irritation even in the ocular mucous membrane irritation evaluation test, a patch test. Therefore, these results suggest that Coptis chinensis has antimicrobial, antifungal and safety on human body and can be applied to the development of materials for cosmetic and residential environment industries.
Kim, Nan-Jin;Ryoo, Hyun-Mo;Kim, Hyun-Jung;Kim, Young-Jin;Nam, Soon-Hyeun
Journal of the korean academy of Pediatric Dentistry
/
v.28
no.2
/
pp.238-246
/
2001
Bone morphogenetic proteins(BMPs), members of the transforming growth factor $\beta$(TGF-$\beta$) superfamily were first identified as the factors that induce ectopic bone formation in vivo, when implanted into muscular tissue. Especially BMP-2 inhibits terminal differentiation of C2C12 myoblasts and converts them into osteoblast lineage cells. In the molecular mechanism of the signal transduction of TGF-$\beta$ and related factors, intracellular signaling proteins were identified as Smad. In previous study, it has been reported that Smad 1 and Smad 5, which belong to the R-Smad family mediate BMP signaling, were involved in the induction of osteoblast differentiation in C2C12 cells. To understnad the role of Smads involved in osteogenic transdifferentiation in C2C12 cell, in present study, after we stably transfected C2C12 cells with each. Smad(Smad 1,Smad 5) expression vector, cultured for 3 days and stained for alkaline phophatase activity. ALP activity positive cells appeared in the Smad 1, Smad 5 stably transfected cell even in the abscence of BMP. After transiently co-transfected C2C12 cells with each Smad expression vector and ALP promoter, it was examined that Smad 1 and Smad 5 expression vector had increased about 2 fold ALP promoter activity in the abscence of BMP. These result suggested that both Smad 1 and Smad 5 were involved in the intracellular BMP signals which induce osteoblast differentiation in C2C12 cells. The effect of BMP on C2C12 cells with Smad 1, Smad 5 transfected were studied by using northern blot analysis. the treatment of BMP upregulated ALP mRNA level in three groups, especially upregulation of ALP was larger in Smad 1, Smad 5 transfected cell than control group. Pretreatment with cycloheximide($10{\mu}g/ml$), a protein synthesis inhibitor resulted in blocking the ALP gene expression even in BMP(100ng/ml) treated cell. These results suggested that Smad increased the level of ALP mRNA via the synthesis of a certain transcriptional regulatory protein.
Ki, Shin-Young;Park, Sung-Woo;Lee, Myung-Ran;Kim, Eun-Young;Uh, Soo-Taek;Kim, Yong-Hoon;Park, Choon-Sik;Lee, Hi-Bal
Tuberculosis and Respiratory Diseases
/
v.45
no.4
/
pp.835-845
/
1998
Background: Silica-induced lung diseases is characterized by the accumulation of inflammatory cells at early stage and fibrosis in pulmonary parenchyma and interstitium at late stage. As a consequence of inflammation, silicosis is accompanied with the expansion of interstitial collagen and the formation of fibrotic nodule. In this process, several kinds of lung cells produce cytokines which can amplify and modulate pulmonary fibrosis. The alveolar macrophage is a potent source of proflammatory cytokines and growth factor. But in the process of silicotic inflammation and fibrosis, there are many changes of the kinetics in cytokine network. And the sources of cytokines in each phase are not well known. Method: 2.5 mg of silica was instillated into the lung of C57BL/6J mice. After intratracheal instillation of silica, the lungs were removed for imunohistochemical stain at 1, 2, 7 day, 2, 4, 8, 12 week, respectively. We investigated the expression of IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ in lung tissue. Results: 1) The expression of IL-6 increased from 1 day after exposure to 8 weeks in vascular endothelium. Also peribronchial area were stained for IL-6 from 7 days and reached the peak level for 4 weeks. 2) The IL-1 $\beta$ was expressed weakly at the alveolar and peribronchial area through 12 weeks. 3) The TNF-$\alpha$ expressed strongly at alveolar and bronchial epithelia during early stage and maintained for 12 weeks. 4) TGF-$\beta$ was expressed strongly at bronchial epithelia and peribronchial area after 1 week and the strongest at 8 weeks. Conclusion: The results above suggests IL-6, TNF-$\alpha$ appear to be a early inflammatory response in silica induced lung fibrosis and TGF-$\beta$ play a major role in the maintenance and modulation of fibrosis in lung tissue. And the regulation of TNF-$\alpha$ production will be a key role in modultion of silica-induced fibrosis.
Lee, Hee Ju;Lee, Sang Gyu;Kim, Sung Kyeom;Choi, Chang Sun;Kim, Sang Beom
Journal of Bio-Environment Control
/
v.25
no.3
/
pp.162-167
/
2016
This study was carried out to investigate proper stem cutting position and air temperature to regenerate roots from cuttings of bitter gourd. 'NS454' (NS) and 'Dragon' (DR) cultivars were tested and the cutting position was cut at the 3rd node in the stem (treatment I) and cut at the central part (stem segment) between 3rd leaf and 4th leaf in the stem (treatment II). The air temperature was maintained at 18, 23, 28 and $35^{\circ}C$ in the growth chambers, respectively. The photosynthetic photon flux at the ground of a chamber was maintained in approximately $150-200{\mu}mol\;m^{-2}s^{-1}$ during the 16-hour photoperiod. The relative humidity in the chambers was maintained over 85%. After 10 days of cuttings, regardless of cutting position, DR and NS cultivars showed 100% the survival rate in the $18^{\circ}C$ or $23^{\circ}C$ treatments, however, most of plants $18^{\circ}C$ treatment had not rooting. In the $28^{\circ}C$ treatment, regardless of cutting position, DR and NS cultivars showed 100% of the rooting rate. In the $23^{\circ}C$ treatment, for DR cultivar, the node cutting showed 90% of rooting rate and the stem segment cutting showed 40% rooting rate. For NS cultivar, the node cutting showed 50% of rooting rate and the stem segment cutting showed 40% rooting rate. DR and NS cultivars have high rooting rate, while the rooting rate remarkably decreased in the $35^{\circ}C$ treatment. For DR cultivar, the stem segment cutting showed 5.3 of the highest rooting number and the node cutting showed 2.7 rooting number in the $28^{\circ}C$ treatment. NS cultivar was not significantly different the rooting regardless of cutting position. The results suggested that cutting at the stem segment cutting of bitter gourd is an effective cutting method for increasing survival rate and the air temperature should be maintained at $28^{\circ}C$.
Park, Jong Woo;Kim, Jinse;Park, Seok Ho;Choi, Dong Soo;Choi, Seung Ryul;Kim, Yong Hoon;Lee, Soo Jang;Park, Chun Wan;Lee, Jung Soo
Food Science and Preservation
/
v.23
no.5
/
pp.614-622
/
2016
This study was carried out to investigate the effect of modified-atmosphere packaging (MAP) on the quality change of "Tabor" tomatoes during long-term exportation periods. Hydroponics tomatoes were harvested at the turning stage, sorted, and box packed and then packaged in nylon film with a pallet. The packaged pallet was filled with a gas composition (5% $O_2$, 1% $CO_2$, and 94% $N_2$) and stored at $10^{\circ}C$ for three weeks. The quality changes in weight loss, firmness, color, acidity, soluble solids, and microorganism growth were measured every 7 day interval. During the initial storage, the pallet-scale MAP showed slightly higher weight loss and firmness changes when compared to the conventional pallet. The total color change (${\Delta}E$) during ripening was delayed 10% under MAP storage. Acidity, soluble solids and phenolic compound contents decreased with increases in storage time regardless of the storage method; however, the quality changes of tomatoes were delayed in the MAP pallet. Furthermore, the decay rate of the pallet-scale MAP stored for 14 days was less than that of the conventional pallet, and the number of microorganisms was approximately 30% lower in the pallet-scale MAP, showing a positive effect on marketability. These results suggested that the pallet-scale MAP of tomatoes could ensure higher quality and longer storage periods than conventional pallet storage.
Pae, Ah-Ran;Won, Hyun-Du;Lee, Richard Sung-Bok;Kim, Hyeong-Seob;Woo, Yi-Hyung
The Journal of Korean Academy of Prosthodontics
/
v.49
no.4
/
pp.333-340
/
2011
Purpose: The aim of this study was to study the effect of hydroxyapatite (HA) coating crystallinity on the proliferation and differentiation of human osteosarcoma cells. Materials and methods: Surface roughness of the titanium disks increased by anodizing treatment and then HA was coated using ion beam-assisted deposition (IBAD). HA coating was crystallized by heat-treated at different temperature ($100^{\circ}C$, $300^{\circ}C$, $500^{\circ}C$, $800^{\circ}C$). According to the temperature, disks were divided into four groups (HA100, HA300, HA500, HA800). With the temperature, crystallinity of the HA coating was different. Anodized disks were used as control group. The physical properties of the disk surface were evaluated by surface roughness tests, XRD tests and SEM. The effect of the crystallinity of HA coating on HOS cells was studied in proliferation and differentiation. HOS cells were cultured on the disks and evaluated after 1, 3, 5, and 7 days. Growth and differentiation kinetics were subsequently investigated by evaluating cell proliferation and alkaline phosphatase activity. Results: Regardless of the heat-treated temperature, there is no difference on the surface roughness. Crystallinity of the HA was appeared in the groups of HA500, HA800. HOS cells proliferation, ALP activity were higher in HA500 and HA800 group than HA100 and HA300. Conclusion: Within the results of this limited study, heat treatment at $500^{\circ}C$ of HA coating produced by IBAD has shown greater effect on proliferation and differentiation of HOS cells. It is considered that further in vivo study will be necessary.
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