Milk and dairy products are not only excellent foods for humans, providing plentiful varied nutrients, but are also a good medium for detrimental food-borne pathogens. Although the food safety field has stabilized due to standardization of food processing, such as the hazard analysis critical control point (HACCP), outbreaks and cases caused by food-borne pathogens still occur at high rates. In approximately 30% of cases, the disease-causing pathogenic organism is undetermined. Recently, a biosensor was developed that has a simple and fast response and overcomes the problems of conventional methods such as cultivation, immuno-assay, polymerase chain reaction, and microarray. Due to the high selectivity and sensitivity of optical biosensors, it is a suitable method for the immediate detection of food-borne pathogens in milk and dairy products.
The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa), group 8 (low-temperature preservation at $0^{\circ}C$ under no additional pressure), group 9 (low-temperature preservation at $-5^{\circ}C$ under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$ water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 7 ($0^{\circ}C$/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance). By the results of this study, low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa suggest the possibility for long term preservation of teeth.
The purpose of this study was to evaluate the viability of periodontal ligament cells in rat teeth using slow cryo-preservation method under pressure by means of MTT assay and WST-1 assay. Eighteen teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both sides of the first and second maxillary molars were extracted as atraumatically as possible under Tiletamine anesthesia. The experimental groups were group 1 (Immediate control), group 2 (Cold preservation at $4^{\circ}C$for 1 week), group 3 (Slow freezing), group 4 (Slow freezing under pressure of 3 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$water bath, then MTT assay and WST-1 assay were processed. One way ANOVA and Tukey method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 4 showed significantly higher viability of periodontal ligament cells than group 2 and 3 (p < 0.05), but showed lower viability than immediate control group. By the results of this study, slow cryo-preservation method under pressure suggests the possibility for long term cryo-preservation of the teeth.
Kim, Jae-Jeung;Kim, Joon-Tae;Park, Sung-Woo;Park, Eun-Suk;Kim, Heung-Tae
The Korean Journal of Pesticide Science
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v.7
no.3
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pp.159-168
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2003
With microtiter plate, the assay method was developed for detecting the fungicidal activity of new compounds against spore germination, spore adhesion and mycelial growth of Colletotrichum sp. JC24 cal1Sing red pepper anthracnose. Also, the effects of some commercialized fungicides on fungal development like above mentioned were investigated by measuring the optical density of mycelia grown into wells of microtiter plate. For the standardization of assay method, some factors, such as the treatment of MTT and/or propanol, inodulum density and incubation period, affecting on mycelial optical density were investigated. For obtaining precise and consistent mycelial optical density, it was necessary the treatment of MTT for 12 hrs and propanol for 1 hr. inoculum density adjusted to $1\times10^5$ spores/mL and incubation period for 36 hrs at $25^{\circ}C$. For fungicidal activities, 6 protective fungicides, 6 ones inhibiting sterol biosynthesis, and one inhibiting respiration were used in this study. While mancozeb, chlorothalonil and dithianon among 6 protective fungicides inhibited strongly spore germination, adhesion, and mycelial growth at $6.25{\mu}g/mL$, propineb, iminoctadine and fluazinam inhibited intermediately spore germination and mycelial growth at $100{\mu}g/mL$. Washing above 3 fungicides with new PD broth, their activity against spore adhesion decreased. With hexaconazole, tebuconazole and myclobutanil, the tendency of the activity against fungal differentiation of the early infection stage was similar to the latter group of protective fungicides, showing the decrease of the inhibitory activity against spore adhesion by washing 2 hrs after incubation. However, kresoxim-methyl inhibited spore adhesion distinctly, depending on the applied concentrations. Based on these results, it might be able to assess the fungicidal activity of many compounds against spore germination, adhesion and mycelial growth by the use of microtiter plate in vitro. Using the assay developed in this report, it was possible to investigate the inhibitory activity of some commercialized fungicides, too.
Kim, Jae-Wook;Kim, Eui-Sung;Kim, Jin;Lee, Seung-Jong
Restorative Dentistry and Endodontics
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v.31
no.3
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pp.192-202
/
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.
This study was conducted to standardize the proper ingredient ratios of chinese cabbage kimchi by the sensory evaluation, chemical properties, and functional properties of antimutagenic effect and inhibitory effect on the growth of cancer cells from the ratios obtained from literatures. The standardized ratios of ingredient from the literatures was 13.0 of radish, 2.0 of green onion, 3.5 of red pepper powder, 1.4 of garlic, 0.6 of ginger, 2.2 of anchovy juice, 1.0 of sugar and the final salt concentration 2.7 in the proportion of 100 salted chinese cabbage. The standardized ratio of the ingredients exhibited better overall acceptability and less moldy smell and moldy flavor than any other ratio of ingredient in the sensory evaluation. The standardized kimchi with the above ratios of the ingredients, at final salt concentration of 2.5%, showed high reducing sugar contents and Leuconostoc sp. counts. All juices of the chinese cabbage kimchi showed not only high antimutagenicity against aflatoxin $B_1$ in Salmonella typhimurium TA100 but also strong inhibitory effect on the growth of AGS human gastric adenocarcinoma cells in SRB assay, especially these functional properties were the most effective at each standardized ratio of the ingredients. From the taste, chemical and functional properties, the standardized ratios of ingredients was 13.0 radish, 2.0 green onion, 3.5 red pepper powder, 1.4 garlic, 0.6 ginger. 2.2 anchovy juice, 1.0 sugar and the final salt concentration 2.5 in the proportion of 100 salted chinese cabbage.
Purpose: Commercial enzyme-linked immunosorbent assay (ELISA) kits have been considered less reliable for children than for adults. The aim of this study was to compare four ELISA kits and in-house immunoblotting based on the analysis of anti-H. pylori-IgG antibody reactivity. Methods: A total of 399 serum samples were collected at the GNU Hospital during 1998-1999. All sera were tested using ELISA and immunoblotting. Statistically significant differences were determined by the $x^2$ test. Results: The overall seropositivity rates using GAP IgG, Genedia IgG, HM-CAP, Pyloriset EIA-G, and immunoblotting were 13.0%, 25.1%, 18.3%, 15.8%, and 62.9%, respectively. Immunoblotting showed a higher seropositivity rate than did all four ELISA kits in all age groups. Genedia IgG had the highest seropositivity among the ELISA kits. The seropositivity rate for children aged 13 to 18 months was lowest, and that of children aged 15 years was highest (90.0%). The seropositivity rate for children aged 7 months to 5 years was significantly lower than that for children aged 6 to 15 years among the four ELISA kits (p<0.0001) and immunoblotting (p=0.02). Conclusion: Immunoblotting is the most sensitive test for detection of anti-Helicobacter pylori IgG antibodies among the serological tests in this study. These results emphasize the need for standardization when commercial ELISA tests are used in different nations or in young age groups. Immunoblotting could be a suitable noninvasive assay for serodiagnosis and seroepidemiologic study of H. pylori infection in Korean children.
Lee, Jin Ho;Kim, Min Jeong;Lee, Jae Woong;Kim, Me Riong;Lee, In Hee;Kim, Eun Jee
Journal of Acupuncture Research
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v.31
no.4
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pp.45-55
/
2014
Objectives : The main aim of this study was to assess the comparative efficiency of two preparation types of Hwangryunhaedok decoction(HRHD-D) using distilled and mixed extraction by measuring the index components and indicators of antioxidant and anti-inflammatory effects. Methods : The antioxidant activity was assessed by comparing distilled and mixed extractions of HRHD-D using an ELISA reader. The anti-inflammatory effect was determined by measuring NO amounts in RAW 264.7 cells. The contents were analyzed with high performance liquid chromatography-diode array detector(HPLC-DAD). Results : The electron donating ability of mixed and distilled extractions obtained with 500 ppm DPPH(1,1-diphenyl-2-picrylhydrazyl assay) solution were 57.8 % and 4.2 %, respectively. The total phenolic content of mixed extraction was 6.9 times that of distilled extraction and total flavonoid content was 51.5 times higher. The anti-inflammatory effect was assessed by NO measurement, and was found to increase significantly dependent on concentration in all mixed extract concentrations(25, 50, 100, 200, $400{\mu}g/mL$), but the difference in distilled extraction by concentration was only significant at 200 and $400{\mu}g/mL$. The HPLC analysis results of mixed extract of HRHD-D showed detection of all four main active constituents of HRHD-D. However, they were not detected in the distilled extract of HRHD-D. Conclusions : Mixed extraction with distillation added to decoction of HRHD-D showed better efficacy in antioxidant and anti-inflammatory effects, and ingredient quantities compared to distilled extraction. Further stability and clinical efficacy studies for standardization of mixed extractions are required.
Purpose: The clinical implication of p53 mutation in gastric cancer is still unclear, as shown by the discordant results that continue to be reported in the literature. Materials and Methods: To assess p53 gene mutation, tumor p53 overexpression, and serum anti-p53 antibody, we employed a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis, an immunohistochemistry using monoclonal antibody DO-7, and an enzymelinked immunosorbent assay (ELISA), respectively. Results: Of 169 surgical specimens of gastric cancer, mutation at exon $5\∼8$ of the p53 was identified in 33 ($19.5\%$) and was significantly correlated with lymph node metastasis. Overexpression of p53 was found in 62 specimens ($36.7\%$) and had a significant correlation with tumor differentiation. Serum anti-p53 antibody was positive in 18 patients ($10.7\%$). Twenty-three of the mutated tumors ($69.7\%$) and 39 of the non-mutated tumors ($28.7\%$) displayed immunoreactivity. Twelve of the immunopositive tumors ($19.4\%$) and 6 of the immunonegative tumors produced anti-p53 antibody. These differences were statistically significant (P<0.001 and P=0.005, respectively). There was no significant difference in survival according to the mutation of p53. Conclusion: Mutation and overexpression of p53 can be easily detected by immunohistochemistry. However, standardization of the immunohistochemical staining method, as well as guidelines for interpreting the stained result, will produce concordant results and thereby improve clinical application.
Hwang, Byung Soon;Kim, Ji Yeong;Kwon, Su Hyun;Kim, Gi-Chang;Kang, Hae Ju;Rhee, Ju Hee;Hwang, In Guk
The Korean Journal of Food And Nutrition
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v.33
no.5
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pp.532-543
/
2020
This experiment was conducted to assess the high antioxidant activity varieties of Chinese cabbage (Brassica rapa L. ssp. pekinensis) from the 55 accessions. The antioxidant activity of Chinese cabbage were determined by the TPC, TFC, DPPH, ABTS, and chlorophyll, carotenoid contents. The TPC and TFC showed a range of 1.21~4.61 mg GAE/g DW, 0.18~3.09 mg CE/g DW. The DPPH and ABTS assay were in the range of 0.65~4.36 and 1.42~6.91 mg ascorbic acid equivalent (ASCE)/g DW, respectively. The UPLC analysis was performed quantitatively to identify chlorophyll and carotenoid in the Chinese cabbage extract. The levels of the total chlorophyll and total carotenoid were 86.60~1,235.91, and 75.86~490.11 ㎍/g, respectively. The comprehensive differences in the total and individual chlorophyll contents have also been observed among different varieties. These results will be valuable as basic data for the standardization of Chinese cabbage.
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