• Restorative Dentistry and Endodontics
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• v.31 no.5
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• pp.398-408
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• 2006

The purpose of this study was to verify the usefulness of MTT analysis as a tool of measurement of the periodontal ligament cell viability from the extracted rat molar. A total of 80 Sprague-Dawley white female rat of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted under Ketamine anesthesia. Twenty-four teeth of each group (divided as five groups depending upon the time-lapse after extraction such as Immediate, 10, 20, 40 and 60 minutes) were immersed in $200{\mu}l$ of MTT solution (0.5 mg/ml) and processed for optical density measurements. Another 10 teeth of each group were treated as same as above and sectioned at $10{\mu}m$ for microscopic examination. All measurements values were divided by the value of hematoxylin-eosin staining which represented the volume of each corresponding samples. Immediate and 10 minute groups showed highest MTT values followed by 20, 40, and 60 minutes consecutively. Statistical significance (p<0.05) existed between all groups except in immediate versus 10 minute groups and 40 versus 60 minutes. Histological findings also showed similar findings with MTT results in crystal shape and crystal numbers between the experimental groups. These data indicate that in vivo MTT analysis nay be of value for evaluation of the periodontal ligament cell viability without time- consuming cell culturing processes.

• Korean Journal of Food Science and Technology
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• v.31 no.4
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• pp.957-963
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• 1999

Effects of cryoprotectants on protein denaturation of soybean curd, tofu, during frozen storage were examined. A whole-coagulated non-press tofu was prepared by adding 2% of isolated soybean protein to soy milk in order to prevent loss of added cryoprotectants. The cryoprotectants added were glocose, glycerol, sorbitol, propylene glycol, and tripolyphosphate. The texture characteristics of soybean curds before and after frozen storage were measured by sensory evaluation and Texture analyzer, and the results were evaluated by response surface methodology (RSM). Glucose, glycerol, sorbitol, and sodium tripolyphosphate were effective as single cryoprotectant, and the mixtures of glucose and sodium tripolyphosphate, and sorbitol and propylene glycol were also effective in minimizing textural change during freezing. Overall, the mixture of cryoprotectants were more effective than single cryoprotectant. According to the RSM, the maximum effect of cryoprotectants in minimizing textural changes during freezing was obtained with the mixture of 2.1% glucose, 6.7% glycerol, 2.1% sorbitol, 0.4% propylene glycol, and 0.3% sodium tripolyphosphate. However, considering the sensory acceptability, the optimum use of cryoprotectants in frozen tofu was 1% glucose, 2% glycerol, 1% sorbitol, 0.2% propylene glycol, and 0.5% sodium tripolyphosphate.

• Restorative Dentistry and Endodontics
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• v.31 no.3
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• pp.192-202
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• 2006

The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen $(-196^{\circ}C)\;with\;4^{\circ}C$ cold preservation group. A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia. Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium) Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in $Viaspan^(R)$). Group 4 (10% DMSO in $Viaspan^(R)$) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium) , Group 6 ($Viaspan^(R)$) at $4^{\circ}C$ for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95 % level of confidence, Another 2 teeth of each group were treated as the same manner and frozen sections $10{\mu}m$ thick for microscopic observation. The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P<0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results. In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.

• Clinical and Experimental Pediatrics
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• v.51 no.10
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• pp.1112-1117
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• 2008

Purpose : We intended to observe cell death and apoptotic changes in neurons in organotypic hippocampal slice cultures following oxygen-glucose deprivation (OGD), using propidium iodide (PI) uptake, Fluoro-Jade (FJ) staining, TUNEL staining and immunofluorescent staining for caspase-3. Methods : The hippocampus of 7-day-old rats was cut into $350{\mu}m$ slices. The slices were cultured for 10 d (date in vitro, DIV 10) and and exposed to OGD for 60 min at DIV 10. They were then incubated for reperfusion under normoxic conditions for an additional 48 h. Fluorescence of PI uptake was observed at predetermined intervals, and the cell death percentage was recorded. At 24 h following OGD, the slices were Cryo-cut into $15{\mu}m$ thicknesses, and Fluoro-Jade staining, TUNEL staining, and immunofluorescence staining for caspase-3 were performed. Results : 1) PI uptake was restricted to the pyramidal cell layer and DG in the slices after OGD. The fluorescent intensities of PI increased from 6 to 48 h during the reperfusion stage. The cell death percentage significantly increased time-dependently in CA1 and DG following OGD (P<0.05). 2) At 24 h after OGD, many FJ positive cells were detected in CA1 and DG. Some neurons had distinct nuclei and processes while others had fragmented nuclei and disrupted processes in CA1. TUNEL and immunofluorescent staining for caspase-3 showed increased expression of TUNEL labeling and caspase-3 in CA1 and DG at 24 h after OGD. Conclusion : The numerous dead cells in the slice cultures after OGD tended to display apoptotic changes mediated by the activation of caspase-3.

• International Journal of Highway Engineering
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• v.9 no.4
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• pp.117-127
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• 2007

The exposed aggregate PCC(EAP) pavements have been successfully used in Europe and Japan as low-noise pavements. Coarse aggregate are exposed on the pavement surface texture of EAP by removing mortar of surface. The pavement surface texture should maintain not only low-noise characteristic but also adequate skid resistance level during the performance period. Skid resistance decreased with wearing and polishing of tire and pavement surface due to the repetition of tire-pavement contact. Since the tires mainly contact the exposed coarse aggregate, the shape and rock type of coarse aggregate significantly influence wearing and polishing of EAP pavements. The test for resistance to abrasion coarse aggregate by use of the Los Angeles machine(KS F 2508) and the method of test for resistance to abrasion coarse aggregate by use of the Accelerated Polishing Machine(ASTM D 3319-90) are generally used to evaluate polishing characteristics of aggregate. In this study, polishing of coarse aggregate of different five rock types were evaluated by KS F 2508(LA abrasion test) and ASTM D 3319-90(PSV method). The results of LA abrasion test and PSV method were contrary to each other. Since LA abrasion test is estimated the quantity of abrasion by the impact of aggregate, it may not be adequate to evaluate the polishing of aggregate by the repetition of tire. In the case of PSV method, the resistance of polishing is estimated the skid resistance variation of polished aggregate after repetition of tire. The PSV method is adequate for the evaluation on polishing of coarse aggregate. From the test results of PSV method, it was founded that rock type, specific gravity, coarse aggregate angularity, flat or elongated particles in coarse aggregate are significant to the resistance characteristic of coarse aggregate.