• Restorative Dentistry and Endodontics
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• v.21 no.1
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• pp.1-18
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• 1996

There are many advantages when using IIF and DNA probe methods over anaerobic culture method in that they are time-and effort-saving, more precise and more sensitive. Furthermore, in IIF and DNA probe methods, the detection is possible only with small amount of bacteria, the quantitative analysis is possible, and the cell viability is not necessary. The purpose of this study is to observe the incidence of P.endodontalis by carrying out anaerobic culture, IIF and colony lift using DNA probe method respectively, and to compare these 3 methods in terms of effectiveness and sensitivity in order to identify the most effective detection method. 30 teeth with at least one clinical symptoms, with single canal, and with pulp necrosis were sampled. For sampling bacteria, access cavity was prepared after disinfecting tooth and its surroundings. Then the paper point was inserted up to the periapical area, leave there for a while, and finally it was placed into PRAS Ringer's sol. and PBS sol. In anaerobic culture method, P.endodontalis was identified by biochemical tests after subculturing black and brown colonies which were produced after 7 days of incubation on BAP and Brucella BAP in anaerobic chamber. To identify P.endodontalis in IIF method, species-specific polyclonal rabbit-antisera of P.endodontalis(ATCC 35406) was reacted with sampled PBS sol. dispensed onto glass slide, and then P.endodontalis was examined by phase contrast microscopy after incubating with Goat anti-rabbit lgG conjugated to Fluorescein isothiocyanate. For colony lift using DNA probe method, membranes were laid over colonies on the surface of BAP and were hybridized with cloned DNA probe of P.endodontalis. The existence of P.endodontalis was then identified by the methods of chemiluminescent detection and color metric detection. Black colony was found in 11 teeth out of 30 teeth and P.endodontalis was detected in 6 teeth (20 %) by anaerobic culture method, 16 teeth (53 %) by IIF method, and 7 teeth (23 %) by DNA probe method. IIF method is significantly better in detecting P.endodontalis than DNA probe method and anaerobic culture method. There was no significant differences between DNA probe method and anaerobic culture method. There was significant correlation between the formation of black colony and the existence of P.endodontalis. The probability of detecting P.endodontalis when black colony being present is 2.89 times higher than when not being present. There was significant relationship between the foul odor of clinical symptoms and P.endodontalis. The sensitivity of existing P.endodontalis when foul odor being present was 93.75 %, while the specificity of not existing P.endodontalis when foul odor not being present was 28.57 %. These results suggested that the probes of P.endodontalis will be used to decide the method and prognosis in endodontic treatments.

• Korean Journal of Food Science and Technology
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• v.39 no.1
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• pp.66-70
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• 2007

Lactobacillus fermentum YL-3 was encapsulated to increase acid tolerance and its total viability. After micro-encapsulation of L. fermentum YL-3 cells with sodium alginate and soybean oil, the morphology of the microcapsule was observed using confocal laser scanning microscopy (CLSM) after staining with pyronin Y and fluorescein isothiocyanate. The sizes of the microcapsules were 120-126 ${\mu}m$, 444-486 ${\mu}m$ and 401-463 ${\mu}m$ when manufactured at pH 2, 3 and 4, respectively. The microcapsule could hold live cells of L. fermentum YL-3 up to $1.2{\times}10^{7}$, $8.1{\times}10^{7}$ and $1.1{\times}10^{8}$ CFU/mL at pH 2, 3 and 4, respectively. The acid tolerance and preservative ability of L. fermentum YL-3 in microcapsule and macrocapsule at $4^{\circ}C$ and $25^{\circ}C$ were tested. L. fermentum YL-3 cells were evenly located in the alginate capsule matrix structure and the firmness of microcapsule was highest at pH 2. Micro-encapsulation showed the most effective acid tolerance at pH 2.0 and preservation of viability at $4^{\circ}C$. However, at $25^{\circ}C$, the macrocapsules showed more effective cell protection than the microcapsules. The application range for microcapsules could be wider than for macrocapsules in the food industry.

• Korean Journal of Environmental Agriculture
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• v.29 no.3
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• pp.282-286
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• 2010

The present studies were carried out for three years from 2007 until 2009 to control nematode and to increase yield of cucumber by crashed-rape seed application at polyethylene film house. Crashed-rape seed has a lot of glucosinolate. Myrosinase decompose gulcosinolate into isothiocyanate and thiocyanate when crashedrape seed go to decay at soil. Those chemical compounds act on poison to nematode at soil. When the crashedrape seed treated at soil, an amount of thiocyanate at soil was risen up. Thiocyanate of plot treated with 200 and 400 kg crashed-rape seed per 10a was 30 and 40 mg/kg, respectively. Nematode(meloidogyne spp) population at soil was 13 to 17 nematodes per dried soil 300g. Yield of cucumber increased 6 to 15 percent to be compared with control. While, Nematode(meloidogyne spp) population of control plot were 463 nematodes per dried soil 300 g. This level was much higher than 150 nematodes which can be brought about injury to plant. Even if the more an amount of crashed-rape seed application, the higher yield of cucumber and control effect of nematode. Consider economical efficiency, 200 kg of crashed-rape seed per 10a was the most effective. Therefore, we suggest applying 200 kg of crashed-rape seed per 10a to control soil nematode when culture cucumber at plastic film house.

• The Korean Journal of Nuclear Medicine
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• v.34 no.2
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• pp.144-153
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• 2000

Purpose: The purpose of this study was to evaluate the relationship between radiation-induced activation of DNA repair genes and radiation induced apoptosis in A431 cell line. Materials and Methods: Five and 25 Gys of gamma radiation were given to A431 cells by a Cs-137 cell irradiator. Apoptosis was evaluated by flow cytometry using annexin V-fluorescein isothiocyanate and propidium iodide staining. The expression of DNA repair genes was evaluated by both Northern and Western blot analyses. Results: The number of apoptotic cells increased with the increased radiation dose. It increased most significantly at 12 hours after irradiation. Expression of p53, p21, and hRAD50 reached the highest level at 12 hours after 5 Gy irradiation. In response to 25 Gy irradiation, hRAD50 and p21 were expressed maximally at 12 hours, but p53 and GADD45 genes showed the highest expression level after 12 hours. Conclusion: Induction of apoptosis and DNA repair by ionizing radiation were closely correlated. The peak time of inducing apoptosis and DNA repair was 12 hours in this study model. hRAD50, a recently discovered DNA repair gene, was also associated with radiation-induced apoptosis.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.10
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• pp.5291-5298
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• 2012

Beta-asarone is one of the main bioactive constituents in traditional Chinese medicine Acorus calamu. Previous studies have shown that it has antifungal and anthelmintic activities. However, little is known about its anticancer effects. This study aimed to determine inhibitory effects on LoVo colon cancer cell proliferation and to clarify the underlying mechanisms in vitro and in vivo. Dose-response and time-course anti-proliferation effects were examined by MTT assay. Our results demonstrated that LoVo cell viability showed dose- and time-dependence on ${\beta}$-asarone. We further assessed anti-proliferation effects as ${\beta}$-asarone-induced apoptosis by annexin V-fluorescein isothiocyanate/propidium iodide assay usinga flow cytometer and observed characteristic nuclear fragmentation and chromatin condensation of apoptosis by microscopy. Moreover, we found the apoptosis to be induced through the mitochondrial/caspase pathway by decreasing mitochondrial membrane potential (MMP) and reducing the Bcl-2-to-Bax ratio, in addition to activating the caspase-9 and caspase-3 cascades. Additionally, the apoptosis could be inhibited by a pan-caspase inhibitor, carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK). When nude mice bearing LoVo tumor xenografts were treated with ${\beta}$-asarone, tumor volumes were reduced and terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) assays of excised tissue also demonstrated apoptotic changes. Taken together, these findings for the first time provide evidence that ${\beta}$-asarone can suppress the growth of colon cancer and the induced apoptosis is possibly mediated through mitochondria/caspase pathways.

• Journal of Life Science
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• v.21 no.6
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• pp.851-857
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• 2011

Sulforaphane (SFN) is an isothiocyanate, isolated from glucoraphanin in broccoli and other cruciferaous vegetables. Recent studies have revealed that SFN induces anti-proliferation and apoptosis by cell cycle arrest in various cancer cells. In this study, we investigated the effect of SFN induced apoptosis in chondrosarcoma HTB-94 cells. SFN caused suppression of proliferation and apoptosis in a dose-dependent manner as determined by cell phenotype, MTT assay and FACS analysis in HTB-94 cells. Treatment of SFN led to caspase-3 activation and p53 accumulation as determined by Western blot analysis. Also, SFN significantly induced DNA fragmentation and nuclear degradation though activation of caspase-3, as detected by DNA electrophoresis and immunostaining, respectively. Our results indicate that SFN-induced apoptosis was regulated by p53 and caspase-3 dependent pathways. Furthermore, SFN may act as a potent anti-proliferation agent, and as a promising candidate for molecular-targeting chemotherapy against human chondrosarcoma cells.

• Journal of Pharmaceutical Investigation
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• v.31 no.4
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• pp.241-256
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• 2001

Alginate microspheres, containing fluorescein isothiocyanate-bovine serum albumin (FITC-BSA) or green fluorescent protein (GFP) were prepared and used as a model drug to develop the oral vaccine delivery system. The alginate microspheres were coated with poly-L-lysine or chitosan. Two methods, w/o-emulsion and spray, were used to prepare alginate microspheres. To optimize preparation conditions, effects of several factors on the particle size and particle morphology of microsphere, and loading efficiency of model antigen were investigated. In both preparation methods, the particle size and the loading efficiency were enhanced when the concentration of sodium alginate increased. In the w/o-emulsion preparation method, as the concentration of Span 80 was increased from 0.5% to 2%, the particle size was decreased, but the loading efficiency was increased. The higher the emulsification speed was, the smaller the particle size and loading efficiency were. The concentration of calcium chloride did not show any effect on the particle size and loading efficiency. In the spray preparation method, the particle size was increased as the nozzle pressure $(from\;1\;kgf/m^2\;to\;3\;kgf/m^2)$ and spray rate was raised. Increasing calcium chloride concentration (<7%) decreased the particle size, in contrast to no effect of calcium chloride concentration on the w/o-emulsion preparation method. Alginate microspheres prepared by two methods were different in the particle size and loading efficiency, the particle size of microspheres prepared by the spray method was about $2-6\;{\mu}m$, larger than that prepared by the w/o emulsion method $(about\;2{\mu}m)$, and the loading efficiency was also higher with spray method. Furthermore, drying process for the microspheres prepared by the spray was simpler and easier, compared with the w/o emulsion preparation. Therefore, the spray method was chosen to prepare alginate microspheres for further experiments. Release pattern of FITC-BSA in alginate microspheres was evaluated in simulated intestinal fluid and PBS (phosphate buffered saline). Dissolution rate of FITC-BSA from alginate/chitosan microsphere was lower than that from alginate microsphere and alginate/poly-L-lysine microsphere. By confocal laser scanning microscope, it was revealed that alginate/FITC-poly-L-lysine microspheres were present in close apposition epithelium of the Peyer's patches of rabbits following inoculation into lumen of intestine, which proved that microspheres could be taken up by Peyer's patch. In conclusion, it is suggested that alginate microsphere prepared by spray method, showing a particle size of & $10\;{\mu}m$ and a high loading efficiency, can be used as a model drug for the development of oral vaccine delivery system.

• Horticultural Science & Technology
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• v.30 no.4
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• pp.409-416
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• 2012

The amount of cellular components including soluble sugars, amino acids, organic acids and glucosinolates (GLS) was investigated during radish root processing to develop a radish beverage. The radish root was divided into two parts, white and green tissue, and processed separately by extracting the juice from the fresh tissue and from the boiled tissue to compare differences in the components content among the preparations. The overall palatability of both the fresh and boiled extracts from the green part of the radish was higher than that of the same extracts from the white part. The sweetness of extract by boiling increased and its pungency decreased, thereby the palatability increased by being compared to the fresh radish extract. The sweetness was affected by sucrose not by glucose or fructose of monosaccharides by showing different sucrose contents according to treatment comparing palatability. Malic acid was identified as primary organic acid, and the content was higher in both the fresh and boiled extracts from the white part than in the extracts from the green part of the radish. The fresh extract from the green part of the radish contained more essential amino acids, such as threonine and valine, and more hydrophilic amino acids including glutamic acid, aspartic acid, and arginine than those of the fresh extract from the white part, suggesting the green fresh part is more palatable than the white fresh part. The main sulfur compound was ethylthiocyanate in radish, and others were butyl isothiocyanate, dimethyl-disulfide, and 4-methylthio-3-butylisothiocyanate. The four GLS were detected much more in the fresh green and fresh white parts of the radish because they evaporated during boiling. The contents of the four sulfur compounds were higher in the white fresh part than in the green fresh part, which is likely the reason the pungency was higher and the palatability was lower in the white fresh part than in the green fresh part of the radish. The ascorbic acid content was higher in the fresh extract compared to the boiled extracts from both the green and white parts. Taken together, these findings indicate that fresh radish extract is superior to obtain in terms of retaining desirable nutritional and functional components for health.

• Korean Journal of Food Science and Technology
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• v.20 no.4
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• pp.511-517
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• 1988

Kimchi, a traditional vegitable fermented food in Korea, was prepared and fermented at $5^{\circ}C$ in order to analyze the taste and flavor compounds. Major flavor components were identified as dimethyl-disulfide, dimethyl-trisulfide, dipropyl-disulfide, 1-butane 1-isothiocyanate, diallyl-disulfide by GC and GC/MS. Some organic acids such as lactic acid and citric acid increased from 1.2mg/100g and 5.4mg/100g in the initial stage to 33.3mg/100g and 44.4mg/100g at the end of fermentation, respectively. Free amino acids in kimchi were found to play an important role for the taste. The contents of total free amino acid increased from 316.3mg/100g to 600mg/100g in the fermented kimchi and glutamic acid, alanine, valine, leucine, lysine and arginine were abundant in kimchi. The results of sensory evaluation showed that the taste of kimchi was closely related to the contents of non-volatile organic acid, free amino acid and pH.

• Journal of the korean academy of Pediatric Dentistry
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• v.39 no.1
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• pp.26-33
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• 2012

The purpose of this study was to find out the effect of dentifrice containing Horseradish (Armoracia rustica) root extracts on improvement of oral hygiene. The clinical evaluation was done participated by 80 adults volunteers who provided a informed consent for their participation. The participants were divided into two groups ; control group (using toothpaste without Horseradish extracts) and experimental group (using toothpaste containing Horseradish extracts). The checklist of the experiment includes the plaque index, gingival index and anti-caries activity. The anti-caries activity was measured by cultivating Streptococcus mutans for 48 hours then estimating the number of increased bacteria using Dentocult SM. The double-blind method was used in this study. The results of this study can be summarized as follows; 1. The plaque formation decreased 28% in control group, 58% in experimental group using dentifrice with Horseradish extracts compared to baseline data. 2. For gingival index, control group and experimental group showed 26% and 40% decrease compared to baseline data, respectively. 3. The score of caries activity test decreased by 4% for control group and 69% for experimental group compared to baseline data. According to the results, a dentifrice containing Horseradish extracts can improve oral hygiene.