The study was carried out to find effects of the saponins that were extracted from red ginseng on the growth of, aflatoxins production by, and protein patterns of Aspergillus flavus NRRL 3357. A. flavus with $10^6$ conidia as grown at $30^{\circ}C$ for seven days on the enriched medium. Mycelial growth and pH changes of medium which cultured the mold, were similar to those of the control group. However, aflatoxin which produced by the mold was less than that of the control in all concentration of the saponin. To be more specific, 0.3 % of the saponin inhibited production of aflatoxin $B_1$ and $G_1$ to the extent of 31.6 and 21% of the control. The protein peaks of A. flavus at the fourth day of the culture were shown high intensity near the level of 14,300 daltons. However, the mold which cultured in the medium containing the saponin showed low intensity of protein than that of the control group on all molecular weight.
The goal of this study is to investigate effects of temperature and co-chaperonin requirement for in vitro protein refolding assisted by E. coli chaperone GroEL under permissive and nonpermissive temperature conditions. In vitro protein refolding of two denatured proteins was kinetically investigated under several conditions in the presence of GroEL. Effects of temperature and GroES-requirement on the process of prevention of protein aggregation and refolding of denatured protein were extensively monitored. We have found that E. coli GroEL chaperone system along with ATP is required for invitro refolding of unfolded polypeptide under nonpermissive temperature of $37^{\circ}C$. However, under permissive condition spontaneous refolding can occur due to lower temperature, which can competes with chaperone-mediated protein refolding via GroEL chaperone system. Thus, GroEL seemed to divert spontaneous refolding pathway of unfolded polypeptide toward chaperone-assisted refolding pathway, which is more efficient protein refolding pathway.
Journal of the Society of Cosmetic Scientists of Korea
/
v.25
no.1
/
pp.23-36
/
1999
Medimin A is a derivative of vitamin A which has been developed by coupling retinoic acid with polyethylene glycol(PEG) to enhance skin permeability and stability. We carried out the collagen synthesis and clinical test to examine the reducing effect of wrinkles by Medimin A. In vitro collagen synthesis was evaluated by quantitative assay of ($^3$H)-proline incorporation into collagenase sensitive protein in fibroblast cultures. Clinical test was evaluated by image analysis of skin replica, visual observation and self-estimated response of volunteers for 10 weeks. Medimin A stimulated about 40% in collagen synthesis. The area of main deep wrinkle on the skin replica was reduced 38.4% with topical application of O/W emulsion containing 0.2% Medimin A. The wrinkles on the eye region was also reduced 25.4%-44.1% by the visual observation and 93% of all volunteers responded that topical application of the O/W emulsion was showed some reducing effect of wrinkles after 10 weeks. From these results, we suggest that Medimin A is a potent anti-wrinkle agent by objective evaluation methods(in vitro collagen synthesis and image analysis of skin replica) and subjective evaluation methods(visual observation and self-estimated response of volunteers).
Proceedings of the Plant Resources Society of Korea Conference
/
2010.10a
/
pp.14-14
/
2010
Vitamin C (ascorbic acid) is an essential component for collagen biosynthesis and also for the proper functioning of the cardiovascular system in humans. Unlike most of the animals, humans lack the ability to synthesize ascorbic acid on their own due to a mutation in the gene encoding the last enzyme of ascorbate biosynthesis. As a result, vitamin C must be obtained from dietary sources like plants. In this study, we have developed two different kinds of transgenic potato plants (Solanumtuberosum L. cv. Taedong Valley) overexpressing strawberry GalUR and mouse GLoase gene under the control of CaMV 35S promoter with increased ascorbic acid levels. Integration of the these genes in the plant genome was confirmed by PCR and Southern blotting. Ascorbic acid(AsA) levels in transgenic tubers were determined by high-performance liquid chromatography(HPLC). The over-expression of these genes resulted in 2-4 folds increase in AsA intransgenic potato and the levels of AsA were positively correlated with increased geneactivity. The transgenic lines with enhanced vitamin C content showed enhanced tolerance to abiotic stresses induced by methyl viologen(MV), NaCl or mannitol as compared to untransformed control plants. The leaf disc senescence assay showed better tolerance in transgenic lines by retaining higher chlorophyll as compared to the untransformed control plants. Present study demonstrated that the over-expression of these gene enhanced the level of AsA in potato tubers and these transgenics performed better under different abiotic stresses as compared to untransformed control. We have also investigated the mechanism of the abiotic stress tolerance upon enhancing the level of the ascorbate in transgenic potato. The transgenic potato plants overexpressing GalUR gene with enhanced accumulation of ascorbate were investigated to analyze the antioxidants activity of enzymes involved in the ascorbate-glutathione cycle and their tolerance mechanism against different abiotic stresses under invitro conditions. Transformed potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed significant increase in the activities of superoxide dismutase(SOD, EC 1.15.1.1), catalase, enzymes of ascorbate-glutathione cycle enzymes such as ascorbate peroxidase(APX, EC 1.11.1.11), dehydroascorbate reductase(DHAR, EC 1.8.5.1), and glutathione reductase(GR, EC 1.8.1.7) as well as the levels of ascorbate, GSH and proline when compared to the untransformed tubers. The increased enzyme activities correlated with their mRNA transcript accumulation in the stressed transgenic tubers. Pronounced differences in redox status were also observed in stressed transgenic potato tubers that showed more tolerance to abiotic stresses when compared to untransformed tubers. From the present study, it is evident that improved to lerance against abiotic stresses in transgenic tubers is due to the increased activity of enzymes involved in the antioxidant system together with enhanced ascorbate accumulated in transformed tubers when compared to untransformed tubers. At moment we also investigating the role of enhanced reduced glutathione level for the maintenance of the methylglyoxal level as it is evident that methylglyoxal is a potent cytotoxic compound produced under the abiotic stress and the maintenance of the methylglyoxal level is important to survive the plant under stress conditions.
Proceedings of the Plant Resources Society of Korea Conference
/
2018.04a
/
pp.67-67
/
2018
Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with $95.4{\pm}0.61%$ of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at $500{\mu}g.ml^{-1}$ showed the total antioxidant of $48.70{\pm}2.90%$, DPPH radical scavenging activity of $37.25{\pm}2.25$, nitric oxide (NO) radical scavenging activity of $54.55{\pm}1.95$ and $H_2O_2$ radical scavenging activity of $43.75{\pm}3.21$. The F41 at $25{\mu}g.ml^{-1}$ displayed antibacterial activity against E. coli ($14.25{\pm}0.2mm$), P. mirabilis ($10.4{\pm}0.6mm$), S. dysenteriae ($18.6{\pm}03mm$), S. paratyphi A ($14.1{\pm}1.1mm$), E. aerogenes ($5.6{\pm}0.4mm$) and S. marcescens ($14.25{\pm}0.2mm$). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) and potent molecules such as 8-[(2E)-2-(3-hydroxybenzylidene)hydrazinyl]-1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, 2-(Dimethylamino)ethyl (1Z)-N-hydroxy-2-(4-morpholinyl)-2-oxoethanimidothioate, Fluorene in the F41, and virtual study revealed that these molecules are likely responsible for the antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells.
Cuticular penetration and detoxication as mechanisms of resistance to the carbamate insecticides in fenobucarb-selected($R_{f}$) and carbofuran-selected($R_{c}$) strains of the brown planthopper (N. lugens Stal) were investigated. Rates of penetration were not significantly different in the susceptible and resistant strains. However, total amount of excretion of the $R_{f}$ and $R_{c}$ strains were much larger than that of the susceptible strain. Fenovucarb and carbofuran were in vivo metabolizd much faster in the $R_{f}$ strain than in the susceptible strain. OSBP(o-sec-butyl phenol) and 3-ketocarbofuran phenol were invitro the major metabolites of fenobucarb and carbofuran in the brown planthopper, respectively. Total amount of the two major metabolites were produced abotu 2 times larger in the $R_{f}$ and $R_{c}$ strains compared to the susceptible strain. OSBP and 3-ketocarbofuran phenol were not so toxic to the brown planthopper ($LD_{50}$ >100 $\mu\textrm{g}$/g hopper). Based on our data, detoxication plays a large role in resistance to fenobucarb and carbofuran in the resistant strain of BPH, although several resistance factors maybe involved.
Effects of wheat arabinoxylan on mouse spleen lymphocytes and peritoneal macrophages were examined in vitro. Among three wheat arabinoxylans (A1: low MW, A2: medium MW, A3: high MW), A3$(50{\sim}100\;{\mu}g/mL)$ increased the viability of spleen lymphocytes up to $114{\sim}125%$ of the control. A1 and A3 $(20\;{\mu}g/mL)$ increased the viability of lipopolysaccharide-treated lymphocytes synergistically. Viability of murine peritoneal macrophages treated with wheat arabinoxylans $(10{\sim}100{\mu}g/mL)$ was increased up to $135{\sim}175%$ of the control. The cytotoxic activity of macrophages against murine lymphocytic leukemic cell increased in the presence of wheat arabinoxylan. Phagocytic index of macrophages treated with wheat arabinozylans $(20\;{\mu}g/mL)$ significantly increased $197{\sim}232%$ compared with the control, and lysosomal phosphatase and myeloperoxidase activities also increased significantly (p<0.05). Treatment of wheat arabinoxylans tended to decrease nitrite production, but significantly stimulated $H_2O_2\;and\;O_2$ productions of macrophages (p<0.05). These results indicate that the immunostimulating effect of wheat arabinoxylan may be closely related with lysosomal enzyme activity and reactive oxygen intermediate production of macrophages.
Journal of The Korean Society of Grassland and Forage Science
/
v.6
no.2
/
pp.78-83
/
1986
This experiment was carried out in order to study the effect of cutting time on the regrowth and the feed composition in native reed. The first growth crops cut in June, July and October and the ratoon crops after cutting cut in October. The samples measured the plant height, leaf blade & sheath-stem ratio, grass yield, feed composition and invitro dry matter digestibility. The results are summarized as follow: 1. As the result of the grass productivity and nutrient yield, it was indicated that suitable cutting time of the native reed was from middle of June to middle of July. The yield of dry matter and in vitro digestible dry matter were much more in the first crops than in the ratoon crops in both of non-fertilizer and fertilizer(P < 0.01), and more in the fertilizer than in non-fertilizer in both of the first and ratoon crops(P < 0.01). 2. The contents of feed composition in native reed of non-fertilizer and fertilizer showed different trends between the first and ratoon crops. In case of the first crops, the contents of C. protein and C.fat were higher in fertilizer than in non-fertilizer and the contents of C. fiber, NDF and ADF were lower in fertilizer than in non-fertilizer, but these contents were not significantly different between non-fertilizer and fertilizer. In the ratoon crops, these contents between non-fertilizer and fertilizer were shown in a opposite manner to those in the first crops.
Forty gentamicin-resistant isolates of Enterococcus faecalis were selected from various clinical materials, determined their antimicrobial susceptibility, and studied there R-plasmid characteristics and polypeptide patterns. All of the isolates were susceptible to vancomycin. The MICs($\mu$/ml) of antimicrobial agents to the isolates were as follows; the MIC of gentamicin was 128 and $\geq$2040, ampicillin 1 and 1, chlorarmphenicol 2 and 8, erythromycin 32 and 256, and vancomycin 1 and 2. E. faecalis HL-1 strain had 8 plasmid DNA elements, HL-2 and HL-3 strains had 6, HL-4 had 7, HL-5 had 4, and HL-6 had 5. The 51.7 Kb of gentamicin resistance plasmid DNA was conjugally transferred from two strains of E. faecalis HL-1 and HL-6 to S. aureus SK 982. The plasmid transfer frequency between S. aureus SK 982 and E. faecalis HL-1 or E. faecalis HL-6 was 6.3$\times10^{-4} and 3.7$\times10^{-5}$, respectively. Plasmid curing ratio after the treatment of ethidium bromide(10$\mu$/ml) to E. faecalis tarnsconjugants R-1 and R-6 were about 51% and 67%, respectively. The tetracycline gene was located in 2.15 Kb plasmid of E. faecalis HL-1, but it was not found in the E. faecalis HL-6 by Southern blot analyses. The antigenic components of E. faecalis HL-1, HL-6, R-1 and R-6 strains were analyzed by SDS-PAGE and immunoblotting. The E. faecalis strains had 7 to 16 polypeptide bands, however their major proteins were 97.8 and 26.8 Kd. At the Immunoblotting, 97.8, 95.8, 74.8, 63.5, 33.7 and 26.8 Kd polypeptides of the strains showed major antigenic activities with patient's sera infected intra-abdominally with an E. faecalis strain.
Kim, Yong-Ki;Kwon, Mi-Kyung;Cho, Weon-Dae;Kim, Tack-Soo;Shim, Hong-Sik;Lee, Yong-Hwon;Lee, Chan-Jung;Lee, Seong-Chan
Research in Plant Disease
/
v.10
no.2
/
pp.105-111
/
2004
Mycelial growth and days required for sclerotial formation(DRSF) in vitro, and disease incidence of infected plants in the field were investigated to evaluate disease resistance of nine garlic cultivars against two species of Sclerotium causing garlic white rot. There were differences among mycelial growth and sclerotial formation of two white rot pathogens on different garlic cultivars in vitro. Mycelial growth showed the highest level on the clove of 'Daeseo'. Sclerotia of two white rot pathogen were formed the earliest on the clove of 'Kodang' and the latest on the clove of 'Namdo'. In field trial all of nine garlic cultivars were highly susceptible to Sclerotium cepivorum and cold-type garlics were less susceptible than warm-type garlics. Meanwhile garlic cultivars used showed differences among resistant responses to Sclerotium sp., forming large sclerotia. All of warm-type garlics showed highly susceptible response, but three cold-type garlics, 'Seosan', 'Danyang' and 'Yechon' showed moderate resistant response and 'Euisung' showed resistant response. To evaluate cultivar resistance of garlics quantitatively disease progress curve was transformed as linear regression equation. Among the intercept (early disease incidence) and the slope (disease infection rate) obtained from the transformed equation, final disease incidence, and area under disease progress curve(AUDPC), AUDPC was the most effective to evaluate disease resistance of garlics to both of white rot pathogens. There was little correlation between mycelial growth and sclerotial formation on garlic cloves in vitro, and epidemiological parameters (early disease incidence, final disease incidence, disease infection rate, AUDPC) obtained from the field trial. Mycelial growth of S. cepivorum was positively correlated with final disease incidence and disease infection rate obtained from the field trial, meanwhile DRSF was negatively correlated with the epidemiological parameters.
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