The responses to antifolales of ToxopLasmc Bondii were investigated by measuring the dihydrorolate redLlctase (DHFR) activity. quantity of DHFR mRNA, and single-strand conformational polymorphism (SSCP) pattern. Pyrimethamine (PYM) and methotrexate (MTX) were tested ds anlifolates. When T. gondii was treated wish PYM, the viability was decreased by the increasing concentration of PYM. DHFR activity tended to increase as the passage proceeded. and the quantity of mRNA expressed was also increased according to passages. The viability of 7. gonnii was decreased by the increasing concentration of MTX, but it was maintained over 40% up to $100{\;}{\mu\textrm{m}}$ MTX. DHFR activity was 77.4% in the 1st passage ($1{\;}{\mu\textrm{m}}$). 82.2% in the 4th passage ($10{\;}{\mu\textrm{m}}$), and 141.3% in the 7th passage ($100{\;}{\mu\textrm{m}}$) But no changes were detected in SSCP pattern of T gondii rxposvd to FYM and MTX. both. These results suggested that the response of T gondii to FYN was rofulalcd by transrriptional level and that, in MTX. the viability of T. gonnii was derived from increasing DHFK activity.
Alstroemeria plants were transformed by using an improved particle-gun-mediated transformation system. Friable embryogenic callus (FEC) induced from the leaves with axil tissues of Alstroemeria plant was used as the target tissue. Also, FEC was transformed with the bar gene was used as a selectable marker. In the case of plasmid pAHC25, 7.5% of the twice-bombarded FEC clumps showed blue foci, whereas the clumps with single bombardment showed only 2.3%. Additionally, a 90° rotation with double bombardment led to a higher frequency (6 times) of luciferase gene expression in PBL9780 than the control treatment. After 8 weeks of bombardment, more than 60 independent transgenic lines were obtained for pAHC25 and nearly 150 independent transgenic lines were obtained for PBL9780, all of which were resistant to PPT and demonstrated either GUS or luciferase activity. Regarding effect of osmotic treatment (0.2 M mannitol) with 7 different periods, the highest transient gene expression was obtained in 8 h before and 16 h after transformation in both pAHC25 and PBL9780. Compared with the control, at least three times more GUS foci and photons were observed in this treatment. With respect to different combinations of mannitol and sorbitol with 8 h before and 16 h after transformation, high numbers of transient and stable transgene expressions were observed in both 0.2 M mannitol and 0.2 M sorbitol used in the osmotic pre-culture. This combination showed the highest transformation efficiency in both pAHC25 (8.5%) and PBL9780 (14.5%). In the control treatment, only 10% of the FEC clumps produced somatic embryos. However, by using 0.2 M mannitol and 0.2 M sorbitol, the frequency of somatic embryos increased to 36.5% (pAHC25) and 22.9% (PBL9780). Of the somatic embryos produced, at least 60% germinated. Approximately 100 somatic embryos from these 210 independent transgenic lines from 2 plasmids developed into shoots, which were then transferred to the greenhouse. PCR analysis confirmed the presence of the bar gene. This is the report on the production of transgenic Alstroemeria plants by using particle bombardment with a high efficiency, thereby providing a new alternative for the transferring of gene of interests in Alstroemeria in the breeding program in the future.
Development of a rapid method possessing the requisite sensitivity and specificity for virus monitoring is necessary for protection of the shellfish-consuming public. Oysters tissue usually contains virus particles in relatively small concentrations along with various other substances that can interfere with detection steps. Therefore, the critical point concerning the detection of viruses in shellfish tissues resides in the processing of samples. The current study demonstrated the possibility of purifying small amounts of virus particles at the interface of a 10/50% sucrose gradient after a single round of sucrose gradient ultracentrifugation. We could detect HAV and poliovirus simultaneously from oyster tissues by using two different sets of primer. Furthermore, the method showed a high level of virus recovery rate (>95%) as determined by plaque assays of the final samples. Taken the advantages of the simple and sensitive methods, it was possible to detect 2 pfu of HAV in 5 g of oyster digestive tissues within 24h.
Twenty one strains strongly producing protease were isolated from Korean traditional Cheonggukjang. Eight strains selected by sensory evaluation on Cheonggukjang prepared with isolated strains were identified with based on biochemical properties a and 16S rDNA sequencing. Identified strains were Bacillus subtilis MB4, and Bacillus amyloliquefaciens A1, A2, B1, MC1, SB2, SC1, and SD1. Protease activities, important strain selection factor, were higher in Cheongjukjang prepared with B. subtilis MB4 (179.6 Unit) and B. amyloliquefaciens SB2 (201.9 Unit) than commercial traditional Cheonggukjang (97.9 Unit). Sensory evaluation revealed Cheonggukjang prepared with B. subtilis MB4 had flavor very similar to commercial traditional Cheonggukjang. Cheonggukjang prepared with B. suhtilis MB4 (0.0006 Pa s) and commercial traditional Cheonggukjang (0.0002 Pa s) revealed lower viscosities than those of Cheonggukjang prepared with B. amyloliquefaciens SB2, MC1, B1, A1, SD1, A2, and SC1 (0.006 to 0.008 Pa s at 1001/s. Results show Cheonggukjang could be prepared using single strain of B. subtilis MB4, maintaining high protease activity and very similar sensory and viscosity qualities with those of commercial traditional Cheonggukjang.
Kim, A-Reum;Jeong, Moon-Jin;Ahn, Yong-Soon;Kim, Mi-Na;Kim, Sung-Im;Lim, Do-Seon
Journal of dental hygiene science
/
v.15
no.2
/
pp.209-219
/
2015
In order to explore an effect of interaction of Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis that are bacteria relevant to periodontal disease on its growth, the bacteria were incubated in trypticase soy hemin menadione broth at $37^{\circ}C$$CO_2$ incubator for 7 days through anaerobic jar by single and co-culture with heat treated dead bacteria under anaerobic gas pack. In order to confirm growth level, absorbance was measured and for confirming colony structure and form, it was observed with scanning electron microscope. In order to confirm an effect on pathogenicity of P. gingivalis, real time reverse transcriptase polymerase chain reaction was implemented for expression analysis for rgpA gene that produces HRgpA which is gingipain. As a result, the following conclusion was obtained. Colony formation of S. gordonii and P. gingivalis was increased by other dead bacteria and in case of F. nucleatum, its colony formation was showed an aspect of being increased by dead bacterium of P. gingivalis but decreased by dead bacterium of S. gordonii. Therefore, it is considered that the strains being used for this study would affect interactively through bacterial cell itself as well as their interaction factor at the time of colony formation.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
/
v.31
no.1
/
pp.22-31
/
2018
Objectives : In this study, we investigated whether Astragali Radix Pharmacocupuncture (ARP) has an effective on the full thickness defect wound healing process of mouse. Methods : A total of 50 mice (ICR mouse, 7 week-old male) were divided into control group and ARP group. A single full thickness skin defect was made on the dorsal side of the each mouse using an 8mm diameter biopsy punch. Control group were treated with 0.2㎖ saline and ARP group were treat with 0.2ml ARP at 8 points around the wound every three days total 4 times during the experimental period. The change in wound size, contraction rate, healing rate, and epithelization rate was measured by digital images taken on days 3, 6, 9, and 13, and evaluated using a digital image analysis program. Tissues were collected for histological analysis, RT-PCR, and Western blot on days 3, 6, 9, and 15. Results : The results are as follows. ARP group accelerated the rate of wound contraction, wound healing and epithelization compared to the control group. ARP group showed the decrease of inflammatory cells in early inflammatory phase compared to the control group. ARP group upregulated PECAM-1 mRNA and protein expression compared to the control group. ARP group inhibited the scar width and area compared to the control group. Conclusions: ARP showed positive effects on wound healing through the inhibition of inflammatory reaction and increase of PECAM-1 expression related to the wound healing process.
Nam, Gun He;Wee, Ji-Hyang;Kim, Sang Yung;Baek, Ji-Young;Kim, Young Min
Journal of Life Science
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v.29
no.10
/
pp.1152-1158
/
2019
Rudbeckia laciniata var. hortensis Bailey is used in home remedy for colic and gastritis in South Korea. Although Rudbeckia laciniata var. hortensis Bailey is used extensively for home remedies, no single study on its efficacy exists. In this study, we investigated the anti-obesity effects of Rudbeckia laciniata var. hortensis Bailey. The anti-obesity effect of a 70% ethanol extract from Rudbeckia laciniata var. hortensis Bailey on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated with an Oil Red O assay, western blot analysis, and mRNA analysis. Compared to the control (only treated with DM), the 70% ethanol extract of Rudbeckia laciniata var. hortensis Bailey significantly inhibited adipocyte differentiation and intracellular triglyceride (TG) levels at a concentration of $100{\mu}g/ml$. To determine how the TG content was reduced, we measured the level of protein and mRNA expression of obesityrelated agents, such as peroxisome proliferators-activated receptor ${\gamma}$ ($PPAR{\gamma}$), CCAAT/enhancer- binding protein ${\alpha}$ ($C/EBP{\alpha}$), AMP-activated protein kinase (AMPK) phosphorylation, LPL, and FAS. As a result, the 70% ethanol extract of Rudbeckia laciniata var. hortensis Bailey significantly increased the expression of AMPK and decreased the expression of genes related to adipogenesis and fat storage, such as $PPAR{\gamma}$, $C/EBP{\alpha}$, LPL, and FAS.
Lee, Ju Yeong;Hwang, Eun Seol;Lee, Jeong-Sub;Kwon, Myunghee;Chung, Hyen Mi;Seo, SungChul
Journal of odor and indoor environment
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v.17
no.4
/
pp.355-361
/
2018
Mold grows more easily when humidity is higher in indoor spaces, and as such is found more often on wetted areas in housing such as walls, toilets, kitchens, and poorly managed spaces. However, there have been few studies that have specifically assessed the level of mold in the indoor spaces of water-damaged housing in the Republic of Korea. We investigated the levels of airborne mold according to the characteristics of water damage types and explored the correlation between the distribution of mold genera and the characteristics of households. Samplings were performed from January 2016 to June 2018 in 97 housing units with water leakage or condensation, or a history of flooding, and in 61 general housing units in the metropolitan and Busan area, respectively. Airborne mold was collected on MEA (Malt extract agar) at flow rate of 100 L/min for 1 min. After collection, the samples were incubated at $25^{\circ}C$ for 120 hours. The cultured samples were counted and corrected using a positive hole conversion table. The samples were then analyzed by single colony culture, DNA extraction, gene amplification, and sequencing. By type of housing, concentrations of airborne mold were highest in flooded housing, followed by water-leaked or highly condensed housings, and then general housing. In more than 50% of water-damaged housing, the level of airborne mold exceeded the guideline of Korea's Ministry of Environment ($500CFU/m^3$). Of particular concern was the fact that the I/O ratio of water-damaged housing was greater than 1, which could indicate that mold damage may occur indoors. The distribution patterns of the fungal species were as follows: Penicillium spp., Cladosporium spp. (14%), Aspergillus spp. (13%) and Alternaria spp. (3%), but significant differences of their levels in indoor spaces were not found. Our findings indicate that high levels of mold damage were found in housing with water damage, and Aspergillus flavus and Penicillium brevicompactum were more dominant in housing with high water activity. Comprehensive management of flooded or water-damaged housing is necessary to reduce fungal exposure.
Bahk, Young Yil;Park, Seo Hye;Kim-Jeon, Myung-Deok;Oh, Sung-Suck;Jung, Haneul;Jun, Hojong;Kim, Kyung-Ae;Park, Jong Myong;Ahn, Seong Kyu;Lee, Jinyoung;Choi, Eun-Jeong;Moon, Bag-Sou;Gong, Young Woo;Kwon, Mun Ju;Kim, Tong-Soo
Parasites, Hosts and Diseases
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v.58
no.5
/
pp.551-558
/
2020
The flaviviruses are small single-stranded RNA viruses that are typically transmitted by mosquitoes or tick vectors and are etiological agents of acute zoonotic infections. The viruses are found around the world and account for significant cases of human diseases. We investigated population of culicine mosquitoes in central region of Korean Peninsula, Incheon Metropolitan City and Hwaseong-si. Aedes vexans nipponii was the most frequently collected mosquitoes (56.5%), followed by Ochlerotatus dorsalis (23.6%), Anopheles spp. (10.9%), and Culex pipiens complex (5.9%). In rural regions of Hwaseong, Aedes vexans nipponii was the highest population (62.9%), followed by Ochlerotatus dorsalis (23.9%) and Anopheles spp. (12.0%). In another rural region of Incheon (habitat of migratory birds), Culex pipiens complex was the highest population (31.4%), followed by Ochlerotatus dorsalis (30.5%), and Aedes vexans vexans (27.5%). Culex pipiens complex was the predominant species in the urban region (84.7%). Culicine mosquitoes were identified at the species level, pooled up to 30 mosquitoes each, and tested for flaviviral RNA using the SYBR Green-based RT-PCR and confirmed by cDNA sequencing. Three of the assayed 2,683 pools (989 pools without Anopheles spp.) were positive for Culex flaviviruses, an insect-specific virus, from Culex pipiens pallens collected at the habitats for migratory birds in Incheon. The maximum likelihood estimation (the estimated number) for Culex pipiens pallens positive for Culex flavivirus was 25. Although viruses responsible for mosquito-borne diseases were not identified, we encourage intensified monitoring and long-term surveillance of both vector and viruses in the interest of global public health.
Journal of Practical Agriculture & Fisheries Research
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v.22
no.1
/
pp.131-142
/
2020
Chrysnathemum stunt viroid (CSVd), a small, single-stranded, infectious RNA, has caused a severe problem in chrysanthemum in the world. In Korea, since CSVd was firstly observed in 1997, the disease has been spread throughout the whole country rapidly. In spite of the seriousness, few methods for control of CSVd have been known without prevention. The use of resistant cultivars is one of the most efficient approaches for overcoming CSVd disease in chrysanthemum cultivation. This study was carried out to identify chrysanthemum genetic resources for the resistance to chrysanthemum stunt viroid (CSVd). A total of 192 commercial cultivars including 167 spray and 25 standard chrysanthemum were screened with CSVd through grafting inoculation method. In most of the inoculated cultivars, typical disease symptoms, stunting of plant height, reduced flower size, and flower color bleaching, were induced. Several cultivars, however, were symptomless or showed delay in symptom expression. Of 192 chrysanthemum cultivars, two cultivars with less than 5% of the average reduction rate of plant height, 'Chiwerel' and 'Jeongheungdaesin', were rated as resistant. And six cultivars less than 20% inculding 'Inga' were rated as moderate resistant. The remaining 184 cultivars with a high level of reduction were rated as susceptible. We expect that these genetic resources can be used in crossbreeding programs for developing CSVd resistant cultivars of chrysanthemum.
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