• Title/Summary/Keyword: ATP-bioluminescence technique

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Rapid Microbiological Assessment Method by using ATP-Bioluminescence in Ginseng Powder (ATP-Bioluminescence를 이용한 인삼분말의 미생물 신속검사법)

  • 곽이성;김천석;송용범;고셩룡
    • Journal of Ginseng Research
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    • v.25 no.3
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    • pp.127-129
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    • 2001
  • Bioluminescence technique was applied to ginseng powders. ATP bioluminescence can be used as a rapid method that can be implemented for microbiological monitoring of contaminated ginseng powders. The RLU (relative light units) of ATP was proportion to bacterial CFU (colony forming units) when in high contaminated ginseng powders ($\geq$ about 1.0$\times$10$^4$CFU/g). However, when in low contaminated ginseng powders ($\times$10$^4$CFU/g), the RLU was not proportion to CFU, respectively.

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Evaluation of Hygienic Status of University Foodservice Operation using ATP bioluminescence Assay (ATP bioluminescence Assay를 이용한 대학 급식시설의 위생상태 평가에 관한 연구)

  • 박영숙
    • Korean journal of food and cookery science
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    • v.16 no.2
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    • pp.195-201
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    • 2000
  • An investigation was conducted to evaluate the hygienic status of university foodservice operation by using conventional swabbing technique plus standard plate count and ATP bioluminescence assay. The results of the study were as follows: 1) For all kitchen boards, knives, feeding trays, and dish towels tested, there was an overall agreement at 84.7% level between the results obtained using ATP bioluminescence and plate count when using a pass/fail cut-off of 3$\times$ control values for ATP assay and 40 CFU(colony forming unit)/㎠ for plate count. 2) The agrement rate between ATP assay and standard plate count was 87.5% for the samples before use, 29.2% for those during use, and 42.7% for those after cleaning and sanitizing. 3) The plate counts of three university foodservice operations for kitchen board, kitchen knife, feeding tray and dish towel were within the acceptable limits when tested before using. However, none of them were within the acceptable limits when tested during using and after cleaning and sanitizing. 4) Above results suggested that an immediate action needs to be taken to reduce the potential danger of cross-contamination and also effective sanitary control methods needs to be developed to improve the sanitary condition.

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Treatment test for bovine mastitis by the determination of ATP based on firefly bioluminescence (Bioluminescence 반응에 의한 ATP측정을 이용한 젖소 유방염 치료에 관한 연구)

  • Kim, Tae-jong;Kim, Jong-bae;Lee, Seong-bae;Jeon, Young-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.393-405
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    • 1989
  • This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

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Verification of the Suitability of the ATP Luminometer as the Monitoring Tool for Surface Hygiene in Foodservices (급식소 식품접촉표면 위생 모니터링 도구로서의 ATP Luminometer 적합성 확인)

  • Kim, Yang-Sook;Moon, Hey-Kyung;Kang, Seong-Il;Nam, Eun-Jeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.11
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    • pp.1719-1723
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    • 2010
  • The purpose of this study was to verify feasibility of using a ATP Luminometer, real-time hygiene monitoring tool for food contact surfaces in foodservices. For this, 54 cutting boards, 70 knives, 21 rubber gloves in 4 institutional foodservices were studied. ATP (RLU: relatively light unit) values by ATP Luminometer were compared with APC (CFU: colony forming unit) of swabbing culture method using aerobic count plates of 3M petrifilm. ATP ranged from 0 RLU/$cm^2$ to 64693 RLU/$cm^2$ on knives, from 0.1 RLU/$cm^2$ to 6743.6 RLU/$cm^2$ on cutting boards and from 31 RLU/$cm^2$ to 465635 RLU/$cm^2$ on the rubber gloves. APC ranged from 0 CFU/$cm^2$ to 166667 CFU/$cm^2$ on knives, from 0 CFU/$cm^2$ to 1000 CFU/$cm^2$ on cutting boards and from 0 CFU/$cm^2$ to 730000 CFU/$cm^2$ on the rubber gloves. To express the degree of association between ATP and APC, a linear regression was performed. There were significant positive correlations found between log RLU and log CFU on the knives (r=0.84, p<0.001), the cutting boards (r=0.79, p<0.001), the rubber gloves (r=0.78, p<0.001). Results of this study showed the possibility that ATP bioluminescence technique can be used as the monitoring tool for surface hygiene in foodservices.

A novel retentive type of dental implant prosthesis: marginal fitness of the cementless double crown type implant prosthesis evaluated by bacterial penetration and viability

  • Hong, Seoung-Jin;Kwon, Kung-Rock;Jang, Eun-Young;Moon, Ji-Hoi
    • 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.

Real-Time Monitoring of Mitochondrial ATP Synthesis and Hydrolysis by Surface Infrared Spectroscopy

  • Yamaguchi, Ryo-Taro;Hirano-Iwata, Ayumi;Aonuma, Yuki;Yoshimura, Yuya;Shinohara, Yasuo;Kimura, Yasuo;Niwano, Michio
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.108-109
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    • 2013
  • Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

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Detection of Multidrug Resistance Using Molecular Nuclear Technique (분자핵의학 기법을 이용한 다약제내성 진단)

  • Lee, Jae-Tae;Ahn, Byeong-Cheol
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.2
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    • pp.180-189
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    • 2004
  • Although the outcome of cancer patients after cytotoxic chemotherapy is related diverse mechanisms, multidrug resistance (MDR) for chemotherapeutic drugs due to cellular P-glycoprotein (Pgp) or multidrug-resistance associated protein (MRP) is most important factor in the chemotherapy failure to cancer. A large number of pharmacologic compounds, including verapamil, quinidine, tamoxifen, cyclosporin A and quinolone derivatives have been reported to overcome MDR. Single photon emission computed tomography (SPECT) and positron emission tomography (PET) are available for the detection of Pgp and MRP-mediated transporter. $^{99m}Tc$-MIBI and other $^{99m}Tc$-radiopharmaceuticals are substrates for Pgp and MRP, and have been used in clinical studies for tumor imaging, and to visualize blockade of PgP-mediated transport after modulation of Pgp pump. Colchicine, verapamil and daunorubicin labeled with $^{11}C$ have been evaluated for the quantification of Pgp-mediated transport with PET in vivo and reported to be feasible substrates with which to image Pgp function in tumors. Leukotrienes are specific substrates for MRP and $N-[^{11}C]acetyl-leukotriene$ E4 provides an opportunity to study MRP function non-invasively in vivo. SPECT and PET pharmaceuticals have successfully used to evaluate pharmacologic effects of MDR modulators. Imaging of MDR and reversal of MDR with bioluminescence in a living animal is also evaluated for future clinical trial. We have described recent advances in molecular imaging of MDR and reviewed recent publications regarding feasibility of SPECT and PET imaging to study the functionality of MDR transporters in vivo.