• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.4
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• pp.288-292
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• 2006

Through statistically designed experiments, lysis agents were optimized to effectively disrupt bacterial cells in a microfluidic device. Most surfactants caused the efficient lysis of Gram-positive microbes, but not of Gram-negative bacteria. A Plackett-Burman design was used to select the components that increase the efficiency of the lysis of the Gram-negative bacteria Escherichia coli. Using this experimental design, both lysozyme and benzalkonium chloride were shown to significantly increase the cell lysis efficiency, and ATP was extracted in proportion to the lysis efficiency. Benzalkonium chloride affected the cell membrane physically, while lysozyme destroyed the cell wall, and the amount of ATP extracted increased through the synergistic interaction of these two components. The two-factor response-surface design method was used to determine the optimum concentrations of lysozyme and benzalkonium chloride, which were found to be 202 and 99 ppm, respectively. The lysis effect was further verified by microscopic observations in the microchannels. These results indicate that Gram-negative cells can be lysed efficiently in a microfluidic device, thereby allowing the rapid detection of bacterial cells using a bioluminescence-based assay of the released ATP.

• Journal of Food Hygiene and Safety
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• v.36 no.1
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• pp.42-50
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• 2021

Along with the rapid growth of the food service industry, food safety requirements and hygiene are increasing in importance in restaurants and hotels. Accordingly, there is a need for quick and practical monitoring techniques to determine hygiene status in the field. In this study, we investigated 5 domestic 5-star hotels specifically, personal hygiene (hands of workers), cooking utensils (knife, cutting board, food storage container, slicing machine blade, ice-maker scoop) and other facilities (refrigerator handle, sink). In addition, we examined the hygiene management status of customer contact points (tongs for buffet, etc.) to derive the correlation between the ATP values as a, a verification method. As a result of our five-hotel survey, we found that cooking utensils and personal hygiene were relatively sanitary compared to other inspection items (cookware 92.2%, personal hygiene 91.4%, facilities and equipment 76.19%, customer contact items 88.6%). According to our ATP-based mothod, kitchen utensils (51 ± 45 RLU/25㎠) were relatively clean compared to other with facilities and equipment (167 ± 123 RLU/25㎠). In the present study, we also evaluated the usefulness of the ATP bioluminescence method for monitoring surface hygiene at hotel restaurants. After correlation analysis of surveillance of hygienic status points and ATP assay, most results showed negative and high correlation (-0.64--0.89). Our ATP assay (92 ± 67 RLU/25㎠) of each item after cleaning showed signigicantly reduced results compared to the ATP assay (1020 ± 1254 RLU/25㎠) for normal status, thereby indicating its suitability as a tool to verify the validity of cleaning. By our results, ATP bioluminescence could be used as an effective tool for visual numerical evaluation of invisible contaminants.

• Journal of Microbiology
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• v.38 no.2
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• pp.74-79
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• 2000

For the demonstration of a novel riboflavin kinase assay method based on the bacterial bioluminescence, partially purified riboflavin kinase was prepared from bovine liver through ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography. Using bacterial luciferase from Photobacterium phosphoreum and the dithionite reduction method, and easy, safe, and fast assay method was established. The optimal temperature, pH, Km values form riboflavin and ATP of boving liver riboflavin kinase determined with this luminescence method were 35$^{\circ}C$, pH 7, 15.3${\mu}$M and 8.3.${\mu}$M, respectively. The detection limit of FMN produced by riboflavin kinase was in the range of 200 pM to 4${\mu}$M which is comparable to the HPLC-fluorescence detection method, while the detection time for each assay was less than 15 sec compared to the HPLC method which requeires at least 10 min for completion.

• Journal of the Korean Dietetic Association
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• v.17 no.2
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• pp.142-160
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• 2011

The purpose of this study was to investigate food sanitation status in elderly welfare facilities and assess the performance of food sanitation practices. Twenty elderly welfare facilities out of 85 located in Seoul with a capacity of fewer than 50 persons participated. The food sanitation status of worktable, kitchen utensils (knives, cutting boards, ladles, spoons), and tableware and bowls were examined by ATP bioluminescence. The results found that the ATP value of knife was the highest. Those of ladles appeared relatively higher than others. Meanwhile, the tableware and bowls, although washed everyday after meals, had the lowest ATP value. This study also conducted a survey on the food sanitation practices of 32 cooking employees in the 20 facilities. Fifty-six percent were in their 40s, and 53% had graduated from high school. More than half (66%) of them had no certification of cooking. Half of the respondents had worked for at least 5 years in food service facilities, and had received food sanitation training. Among them, 31% said they applied food sanitation training while working, and 47% responded the training was very helpful. The foodservice employees demonstrated good food sanitation practices. The results show that food sanitation performance of the workers significantly differed according to their age, education level, total work experience in food service facilities, chef certification, and prior food sanitation experience.

• Journal of Microbiology and Biotechnology
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• v.33 no.11
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• pp.1506-1512
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• 2023

Quantitative analysis of adenosine triphosphate (ATP) has been widely used as a diagnostic tool in the food and medical industries. Particularly, the pathogenesis of a few diseases including inflammatory bowel disease (IBD) is closely related to high ATP concentrations. A bioluminescent D-luciferin/luciferase system, which includes a luciferase (FLuc) from the firefly Photinus pyralis as a key component, is the most commonly used method for the detection and quantification of ATP. Here, instead of isolating FLuc produced in recombinant Escherichia coli, we aimed to develop a whole-cell biocatalyst system that does not require extraction and purification of FLuc. To this end, the gene coding for FLuc was introduced into the genome of probiotic Saccharomyces boulardii using the CRISPR/Cas9-based genome editing system. The linear relationship (r² = 0.9561) between ATP levels and bioluminescence generated from the engineered S. boulardii expressing FLuc was observed in vitro. To explore the feasibility of using the engineered S. boulardii expressing FLuc as a whole-cell biosensor to detect inflammation biomarker (i.e., ATP) in the gut, a colitis mouse model was established using dextran sodium sulfate as a colitogenic compound. Our findings demonstrated that the whole-cell biosensor can detect elevated ATP levels during gut inflammation in mice. Therefore, the simple and powerful method developed herein could be applied for non-invasive IBD diagnosis.

• Journal of Food Hygiene and Safety
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• v.24 no.3
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• pp.277-284
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• 2009

Currently food-borne disease is being increased at outdoor food services including hotels and restaurants. Speedy and convenient practical monitoring techniques to determine hygienic conditions are needed. This study was designed to verify correlation of direct ATP (Adenosine Tri-Phosphate) examination method using ATP bioluminescence and surveillance with check-list by inspector. Hygienic status of personal hygiene (hands), kitchen utensils (knives, chopping boards, kitchen towels, cap openers, food storage containers, and blade of slice machines), facilities and equipments (refrigerator handles, worktables, and sinks) in five major hotels in Seoul were examined. The result of personal hygiene of hotels was relatively better than other inspection items (46.6 points in personal hygiene, 40.2 points in kitchen utensils, 40.3 points in facilities & equipments). In ATP inspection, kitchen utensils and facilities & equipments were relatively clean comparing with personal hands data ($40.8{\pm}6.77\;RLU/cm^2$). After correlation analysis of surveillance in hygienic status points and ATP value, all results showed negative and high correlation. The surveillance data and ATP results investigating personal hygiene, kitchen utensils and facilities & equipments were highly correlated. The ATP examination method which shows real-time identification could be considered as an appropriate method to alternate current check-list dependent safety management in food services including hotels.

• The Journal of Advanced Prosthodontics
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• v.12 no.4
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• pp.233-238
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• 2020

PURPOSE. This study aims to compare the marginal fitness of two types of implant-supported fixed dental prosthesis, i.e., cementless fixation (CL.F) system and cement-retained type. MATERIALS AND METHODS. In each group, ten specimens were assessed. Each specimen comprised implant lab analog, titanium abutment fabricated with a 2-degree tapered axial wall, and zirconia crown. The crown of the CL.F system was retained by frictional force between abutment and relined composite resin. In the cement-retained type, zinc oxide eugenol cement was used to set crown and abutment. All specimens were sterilized with ethylene oxide, immersed in Prevotella intermedia culture in a 50 mL tube, and incubated with rotation. After 48 h, the specimens were washed thoroughly before separating the crown and abutment. The bacteria that penetrated into the crown-abutment interface were collected by washing with 500 µL of sterile saline. The bacterial cell number was quantified using the agar plate count technique. The BacTiter-Glo Microbial Cell Viability Assay Kit was used to measure bacterial adenosine triphosphate (ATP)-bioluminescence, which reflects the bacterial viability. The t-test was performed, and the significance level was set at 5%. RESULTS. The number of penetrating bacterial cells assessed by colony-forming units was approximately 33% lower in the CL.F system than in the cement-retained type (P<.05). ATP-bioluminescence was approximately 41% lower in the CL.F system than in the cement-retained type (P<.05). CONCLUSION. The CL.F system is more resistant to bacterial penetration into the abutment-crown interface than the cement-retained type, thereby indicating a precise marginal fit.