Park Gyung Soo;Yoon Seong Jin;Lee Seung Min;Kim Ae Hyang;Park Soung Yun;Kang Duk Young
Korean Journal of Environmental Biology
/
v.23
no.3
s.59
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pp.293-303
/
2005
Japanese medaka, Oryzias latipes is widely distributed in the North East Asia including Korea, Japan and east China, and commonly used for freshwater toxicity tests and cytotoxicological studies worldwide. In this study, a series of experiments were conducted to identify the potential of the fish as a standard test species for saltwater toxicity evaluation such as marine receiving waters, ocean-dumped materials and sediment pore waters etc. Hatching, growth and mortality rates of the fish were estimated with the wide ranges of salinity from freshwater to seawater (35 psu). Direct exposure of the fertilized eggs in freshwater to the wide ranges of salinity (from 0 to 35 psu) without pre- acclimation to the saltwater revealed no significant differences in hatching rates by salinities (p =0.24). On the other hand, medaka larvae hatched in freshwater and exposed to saltwater directly showed high mortality at > 25 psu treatment groups (p < 0.0001). However, there was no significant difference in mortality of medaka larvae hatched in 13.8 and 14.2 psu at the wide ranges of salinities ($0\~35$ psu). Growth rates of medaka larvae hatched in the above two salinities showed no differences in body length either from 0 to 35 psu treatment groups (p =0.64 for 13.8 psu group and p=0.32 for 14.2 psu group). The number of gill chloride cell in medaka larvae sharply increased when the larvae were exposed to high salinity. Reference tests with zinc chloride revealed 96h $LC_{50}=8.84(7.19\~10.87)mg\;L^{-1}$ using 7~10 day old medaka larvae. These were comparable or better sensitivity in comparison with the other standard test species such as North American sheepshead minnow Cyprinodon variegatus. Based on the results of these experiments, hatching rates and larvalmortality of medaka must be good toxicity parameters for seawater bioassay and the species seems to be a good standard species for both the freshwater and seawater toxicity test.
This study was designed to examine the suitable characteristics of potential probiotic bacteria. Possible probiotic bacteria, including Lactobacillus acidophilus DDS-1, Lb. acidophilus B-3208, Bifidobacterium bifidum KCTC 3357, Lb. plantarum, Leuconostoc mesenteroides ssp. mesenteroides ATCC 8293, and Lactococcus lactis ssp. lactis ATCC 7962 were selected. We then measured their acid and bile tolerances, adhesion properties in the gastrointestinal tract, antimicrobial activity against pathogenic bacteria, and immunomodulation activity. The acid tolerances of Lb. acidophilus DDS-1, Lb. acidophilus B-3208, Lb. plantarum, and Leu. mesenteroides ssp. mesenteroides ATCC 8293, in PBS (pH 2.5) for 2 hr, were high enough that 50% of the inocula survived. The bile tolerances of all bacteria, except Lc. lactis ssp. lactis ATCC 7962, were also observed at a 3% oxgall concentration in MRS broth. The results of the adhesion property assay showed that the total binding affinities of Lb. acidophilus DDS-1, Lb. acidophilus B-3208, and B. bifidum were about three times higher than those of the other bacteria. In testing their antimicrobial activities against pathogens, Lb. acidophilus B-3208, B. bifidum KCTC 3357, and Lb. plantarum inhibited the growth of pathogenic bacteria. For their immunomodulation activity, the cell wall fractions from Lb. acidophilus DDS-1 and Lb. acidophilus B-3208 showed the highest bone marrow cell proliferation activities. However, the cell wall fractions of Lb. acidophilus DDS-1 and B. bifidum, and the cytosol fraction of Lc. lactis ssp. lactis ATCC 7962 showed higher macrophage stimulation activities than those of the other bacteria. Since Lb. acidophilus DDS-1 and Lb. acidophilus B-3208 satisfy the requirements for probiotics, they can be considered suitable probiotic bacteria.
This study was conducted to compare the methods of slide dip and leaf dip techniques in the resistant level determination of spider mites to acaricides used; .and to investigate resistant levels of two-spotted spider mite, Tetranychus urticae Koch to benzomate, dicofol and cyhexatin in Honam region of Korea. Comparisons of the resistant strains and susceptible strain based on $LC_{50}$ values were summarized. Using the slide dip and leaf dip techniques with benzomate acaricide, the Keumchun strain of T. urticae showed 75-and 69-fold resistant levels, respectively, as compared with the Kwangju susceptible strain, but the Hackyo strain was 38-and 35-fold resistant levels, respectively. There was no significant difference (P>0.05) between the slide dip and leaf dip methods. With dicofol acaricide, resistant levels of the Keumchun strain of T. urticae were 71 times by using the slide dip and 74 times by using the leaf dip as resistant as the Kwangju susceptible strain while the Hackyo strain was 24 times by the former method and 23 times by the later method. There was also no significant difference (P>0.05) between both the methods. The slide dip technique is more recommendable than the leaf dip technique for determination of degree of chemical resistance to the spider mites with accuracy and less variation of the results. Using the slide dip method with cyhexatin, the Bia, Keumchun and Hackyo strains of T. urticae were 19-, 18- and 9-fold resistant levels, respectively, over the Kwangju susceptible strain.
The effect of inlay surface treatment on bonding was investigated when resin inlay was bonded to resin-modified glass-ionomer base with resin cement. For the preparation of glass-ionomer base, resin-modified glass-ionomer cement (Fuji II LC, GC Co., Japan) was filled in class I cavities of 7mm in diameter and 2mm in depth made in plastic molds. Eighty eight resin inlay specimens were made with Charisma$^{(R)}$ (Kulzer, Germany) and then randomly assigned to the four different surface treatment conditions: Group I, $50{\mu}m$ aluminium oxide sandblasting and silane treatment ; Group II, silane treatment alone ; Group III, sandblasting alone, and Group IV (control), no surface treatment. After a dentin bonding agent with primer (One-Step$^{TM}$, Bisco Inc., IL., U.S.A.) was applied to bonding surface of resin inlay and base, resin inlay were cemented to glass-ionomer base with a resin cement (Choice$^{TM}$, Bisco Inc., IL., U.S.A.). Shear bond strengths of each specimens were measured using Instron universal testing machine (4202 Instron, lnstron Co., U.S.A.) and fractured surfaces were examined under the stereoscope. Statistical analysis was done with one-way ANOVA and Dunkan's multiple range test. The results were as follows: 1. Sandblasting and silane treatment provided the greatest bond strength(10.56${\pm}$1.95 MPa), and showed a significantly greater bond strength than sandblasting alone or no treatment (p<0.05). 2. Silane treatment provided a significantly greater bond strength(9.77${\pm}$2.04 MPa) than sandblasting alone or no treatment (p<0.05). However, there was no significant difference in bond strength between sandblasting treatment and silane one (p>0.05). 3. Sandblasting alone provided no significant difference in bond strength from no treatment (p>0.05). 4. Stereoscopic examination of fractured surface showed that sandblasting and silane treatment or silane treatment alone had more cohesive failure mode than adhesive failure mode. 5. In relationship between shear bond strength and failure mode, cohesive failure occurred more frequently as bond strength increased.
Environmentally sound control of Japanese lawngrass cutworm, Spodoptera depravata with Korean entomopathogenic nematodes (Heterorhabditis sp. Gyeongsan, Steinernema carpocapsae GSNI , S. glaseri Dongrae, S. longicaudum Nonsan, and S. monticolum Jiri) was evaluated in the laboratory and field. In addition, effect of turf height on the pathogenicity of entomopathogenic nematodes and density of ant (Formica japonica) and spine-tailed earwigs (Forficula scudderi) were investigated in the field. The corrected mortality of 3rd instar of S. depravata larvae was 53.3% to 66.7% 5 days later after treatment of entomopathogenic nematodes. $LC_{50}$ value of S. carpocapsae GSN1 and S. monticolum Jiri against 3rd instar of S. depravata was 6.9 and 3.9, respectively. Pathogenicity of S. carpocapsae GSN1 was different depending on larval stages, i.e., mortality of 2nd instar was the highest representing 73.3% but decreased from 3rd instar. Turf height affected pathogenicity of entomopathogenic nematodes. Mortality of 4th instar of S. depravata was 40.0% by S. carpocapsae GSN1 and 33.3% by S. monticolum Jiri, and 83.3% by fenitrothion, respectively, in the turf height of 14mm, but those were lower in 45mm. The corrected mortality of S. depravata larvae was lower at the entomopathgenic nematode plots than fenitrothion plot in the distribution field of Formica japonica. However, reduction rate of F japonica was higher in fenitrothion by 56.7% compared with 0% in S. carpocapsae GSN1 and 6.7% in S. monticolum Jiri. Turf height influenced control of S. depravate in S. carpocapsae GSN1 and fenitrothion. Control value of S. carpocapsae GSN1 was 62.2% in the turf height of 3-4cm, but not effective at all in 6-8cm and 12-14cm. However, there was not significantly different in fenitrothion plots. Forficula Scudderi was also reduced only in fenitrothion plots. Reduction rate was 100% in 3-4cm, 41.7% in 6-8cm, and 16.7% in 12-14cm, respectively.
A series of experiments were carried out to evaluate the differences in susceptibility of the 13-aphidicidial insecticides in the green peach aphids(Myzus persicae) collected from 13 different localities. The aphids were reared and increased on the potted tobacco plants in the laboratory. The susceptibility to insecticides was compared at the $LC_{50}$ levels with the resistant ratio by a leaf-dip method with a laboratory strain reared for 2 years in the laboratory without exposure to insecticides. The susceptibility to the insecticides was greatly varied with the local strains. The local strains demonstrated relatively high resistance to the insecticides over the laboratory strain at the $LC_{50}$ level; in resistant fold, 2.3 to 519.0 to cypermenthrin, 2.3 to 494.5 to decamethrin, 2.8 to 442.4 to fenvalerate, 2.5 to 170.6 to formothion, 1.5 to 494.5 to decamethrin, 2.8 to 442.4 to fenvalerate, 2.5 to 170.6 to formothion, 1.5 to 231.8 to phosphamidon, 3.1 to 42.1 to monocrotophos, 1.0 to 30.9 to phenthoate+dimethoate, 1.8 to 21.0 to heptanophos, 2.1 to 24.8 to oxydemeton-methyl, 1.0 to 24.9 to thiometon, 1.6 to 4.7 acephate, 0.8 to 4.1 to pirimicarb.
Antifungal compounds from Bacillus polyfermenticus CJ6 were purified using SPE, preparative HPLC, and reverse phase-HPLC. Antifungal compounds from B. polyfermenticus CJ6 were separated into three fractions (8, B, C) using preparative HPLC. LC/MS analysis of antifungal peaks suggested that B. polyfermenticus CJ6 produces lipopeptides; two kinds of iturin A ($C_{14}$, $C_{15}$), three kinds of surfactins ($C_{13}$, $C_{14}$, $C_{15}$), four kinds of fengycin A ($C_{14}$, $C_{15}$, $C_{16}$, $C_{17}$) and two kinds fengycin B ($C_{16}$, $C_{17}$). The antifungal activity of fraction 8, which was presumed as inturin A, was found to be stable after the pH, heat or proteolytic enzyme treatment, but it was unstable at 50-$70^{\circ}C$ for 24 hr. The antifungal activity of fraction B, which presumed as surfactins and fengycin A, was found to be stable after the heat treatment, but it was unstable in the pH 3.0 and after the protease (type I) or ${\alpha}$-chymotrypsin treatment. The antifungal activity of fraction C, which was presumed as fengycin A and B, was found to be stable in the pH 3.0-9.0 range and the heat treatment, but it was unstable with the treatment of protease (type I). The amino acid composition of the purified peaks 8-1 and 8-2 were Asx, Tyr, Gln, Pro, and Ser in a molar ratio of 3:1:1:1:1, which showed the same amino acid composition as iturin. From these results, we confirmed that antifungal compounds from B. polyfermenticus CJ6 most likely belonged to iturin A as well as surfactins and fengycins. As lipopeptides are known to act in a synergistic manner, the antifungal compounds from B. polyfermenticus CJ6 might have potential uses in biotechnology and biopharmaceutical applications.
In this study, the acetylcholinesterase (AChE) inhibition and neuronal cell protective effects of water, 100% methanol and dichlromethane extracts from garlic were investigated. We found that dichloromethane extract of garlic resulted in a dose-dependent manner on AChE inhibition ($IC_{50}$: $36.1{\mu}g/mL$). In cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), cell viabilities of water, 100% methanol and dichlromethane extracts were lower (almost under 40%) than amyloid ${\beta}$ protein ($A{\beta}$)-induced neurotoxicity. Because $A{\beta}$ is also known to increase neuronal cell membrane breakdown, neuronal apoptosis was further confirmed by lactate dehydrogenase (LDH) and neutral red uptake (NRU) assay. Water extract presented relative protection against $A{\beta}$-induced membrane damage in LDH assay. However all garlic extracts showed significant problem with decrease of cell viability in NRU assay, especially at dichloromethan extract. To determine active compounds in column fractions (98:2 fraction) from dichloromethane extract which showed significant AChE inhibitory effect, we performed HPLC and LC-MS analysis. It was supposed that garlic may contain allyl methyl disulfide, diallyl monosulfide, and diallyl disulfide as active compounds.
Shin, Hyuk Soo;Kim, Minwoo;Song, Jerry;Lee, Junseok;Ha, Yoonjeong;Jeon, Young Hee;Kim, Ji Woong;Lee, Yun Ju;Park, Soo Nam
Journal of the Society of Cosmetic Scientists of Korea
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v.44
no.4
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pp.407-417
/
2018
In this study, the antioxidant, cytoprotective and antimicrobial activities of 50% ethanol extract of Polygoni multiflori Radix (PMR) and its ethyl acetate fraction were evaluated to confirm the applicability as a functional ingredient. The activities of the major constituent of PMR were verified and 2, 3, 5, 4′-tetrahydroxystilbene 2-O-${\beta}$-D-glucoside (THSG) was confirmed to be the main component of extract and fraction using HPLC-DAD, LC-EIS-MS analysis. The phenolic and THSG contents of the ethyl acetate fraction were 11.1- and 3.0-folds higher than those of the ethanol extract, respectively. As a result of the DPPH assay and that of luminol dependent chemiluminescence assay in $Fe^{3+}$-EDTA/H2O2 system. the ethylacetate fraction was superior to the ethanol extract in free radical and ROS scavenging activities. Especially, the ethyl acetate fraction and THSG exhibited the similar scavenging activity like L-ascorbic acid in ROS scavenging activity. The ethyl acetate fraction perceived the most potent cytoprotective effect against oxidative damage of erythrocytes induced by photosensitization reaction, followed by the ethanol fraction, THSG and that of (+)-${\alpha}$-tocopherol, which was used as a positive control. Antimicrobial activities were evaluated by disc diffusion and broth microdilution assay against S. aureus, E. coli, P. aeruginosa and C. albicans. In particular, the antibacterial activity of the extract and fraction against S. aureus was superior to that of methyl paraben. Taken together, our results suggest that PMR could be used as a natural ingredient for antioxidant, cytoprotective and antimicrobial activities.
Lung cancer is a type of cancer that has the highest mortality rate. It is mainly classified into small-cell lung cancer (SCLC) and non-small-cell lung cancer (NSCLC). Chemotherapy is used to treat lung cancer, but long-term treatment causes side effects and drug resistances. Curcumin is a bright yellow polyphenol extracted from the root of turmeric. It has biological activities, such as anti-oxidant, anti-cancer, and anti-inflammatory effects. In this study, we observed differential cell death in human lung cancer cells. Based on the results, curcumin at 10, 30, and 50 μM exhibited a dose-dependent inhibition on the cell survival of several lung cancer cells, with minor differential phenotypes. In addition, apoptosis, autophagy, and reactive oxygen species (ROS) regeneration were observed through flow cytometry. Curcumin dose-dependently increased these phenotypes in A549 (NSCLC) and DMS53 (SCLC), which were restored by corresponding inhibitors. Western blotting was performed to measure the level of expression of apoptosis- and autophagy-related proteins. The results indicate that Bax, PARP, pro-caspase-3, and Bcl-2 were dose-dependently regulated by curcumin, with seemingly higher Bax/Bcl-2 ratios in DMS53. In addition, autophagic proteins, p-AKT, p62, and LC3B, were dose-dependently regulated by curcumin. ROS inhibition by diphenyleneiodonium reduced the induction of apoptosis and autophagy generated by curcumin. Taken together, it is suggested that curcumin induces apoptosis and autophagy via ROS generation, leading to cell death, with minor differences between human lung cancer cells.
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