ATP luminescence measurements (using Relative Light Units, RLU) has been used to assess the levels of bacterial contamination on the surfaces of various materials. However, not much is known about their suitability in assessing bacterial contamination on paper surfaces. This study was conducted to evaluate the feasibility of using ATP luminometers in measuring levels of bacteria contamination on paper surfaces. The three ATP luminometers studied were Clean-Q, smart PD-30, and 3M™ Clean-Trace™ LM1 manufactured by different companies. There were some differences in RLU results among the three ATP luminometers when they were tested with different concentrations of Micrococcus luteus cell suspension. 106 - 107 cells were required in order to effectively detect Bacillus subtilis, Escherichia coli, and Micrococcus luteus on the surfaces of A4 printing sheets (100 cm2) when using the three ATP luminometers. The sizes and physical properties of surface areas varied slightly among the swabs used for the three ATP luminometers. Concentration-specific measurements (RLU) of M. luteus taken from the surfaces of six kinds of paper (fine print paper, medium print paper, ground paper, newsprint paper, practice paper, tracing paper) were possible using the smart PD-30 and LM1 ATP luminometers. ATP detection values of M. luteus varied among the six types of paper. The highest ATP detection values were found on the surfaces of tracing paper. If the RLU value is recorded at the level of 1000, this could indicate a very high bacterial contamination level of 105 to 106 CFU/4 cm2.

This study was conducted to obtain basic information for the use of the ATP fluorescence detection method in consideration of the most common and frequent contamination situation that occurs in laboratories dealing with fire blight causing bacterium, Erwinia amylovora. ATP luminescence measurements (Relative Light Unit, RLU) were tested against these pathogen cells (CFU/cm2) which were artificially introduced on the disinfected surface of a bench floor of a biosafety cabinet (Class 2 Type A1), on a part of the disinfected surface of a lab experimental bench, on a part of the disinfected floor, and on a part of the disinfected floor of an acryl chamber for bioaerosol studies in a biosafety laboratory (BSL 2 class) using two different ATP bioluminometers. RLU values were not much increased with the bacterial cells from 2.15 × 102/cm2 to 2.15 × 106/cm2. RLU values varied among the four different surfaces tested. RLU values measured from the same number of bacterial cells differed little between the two different ATP bioluminometers used for this study. RLU values obtained from bacterial cells higher than 2.15 × 107/cm2 indicated the presence of bacterial contamination on the four different surfaces tested. The R2 values obtained based on the correlation data for the RLU values in response to different E. amylovora cell numbers (CFU/ cm2) on the surfaces of the four test spots ranged from 0.9827 to 0.9999.

외식산업의 급속한 성장과 함께 호텔을 비롯한 외식업체에서는 식품안전 요구 및 위생에 대한 중요성 이 증가하고 있다. 이에 현장에서 위생상태를 판단할 수 있는 신속하고 실용적인 모니터링기법이 요구된다. 본 연구는 국내5성급 호텔 5군데를 대상으로 개인위생(작업자의 손), 조리기구(칼, 도마, 식품보관용기, 슬라이스 머신 칼날, 제빙기 스쿱), 시설·설비(냉장고 손잡이, 작업대, 싱크대), 고객 접점항목(뷔페용 집게)에 대한 위생관리 실태를 조사하였고 그에 대한 검증법인 ATP 값의 상관관계를 도출하고자 하였다. 5개 호텔의 위생관리 실태조사 결과, 다른 검사 항목보다 상대적으로 조리기구 및 개인위생 결과가 비교적 위생적으로 관리되고 있었으며(조리기구 92.2%, 개인위생 91.4%, 시설·설비 76.19%, 고객 접점항목 88.6%) 으로 시설·설비는 비교적 미흡한 것으로 나타났다. ATP검사 결과, 조리 기구(51±45 RLU/25 cm2)는 시설·설비(167±123 RLU/25 cm2)보다 비교적 위생적으로 잘 관리되었다. 위생실태 조사 결과 점수와 ATP 값의 상관성 분석을 실시한 결과, 각 호텔 별 작업자 손, 조리기구, 시설·설비의 대부분 음의 상관관계를 가지며 높은 상관성(-0.64 - -0.89) 을 보였다. 또한 이번 연구에서 각 검사 항목의 평소 상태의 ATP 값(1020±1254 RLU/25 cm2)에 비해 세척 후 ATP 값(92±67 RLU/25 cm2)이 현저히 감소되어 세척의 유효성을 확인하는 도구로 적합성을 확인할 수 있었다. 호텔을 비롯한 외식업체에서 수행하는 주관적인 위생실태조사와 ATP 검사법을 병행한다면, 실시간으로 보이지 않는 오염 물질의 객관적인 수치화를 통해 식품사고 발생을 예방하기 위한 효과적인 모니터링 방법이 될 것으로 사료된다.

An investigation was conducted to evaluate the hygienic status of 53 high school foodservice systems in Gyeonggi province by using hygiene management guide checklist, ATP bioluminescence assay of food utensils were conducted during process. The 5 hygiene management guide checklist groups about personal hygiene, cooking facilities control, cross contamination control, cook and storage control, management control were checked by experts and had good grades but there were some inadequate behaviors on observation. Total cleaning levels were inadequate, including hand, rubber gloves, aprons, knives, food tray, machine and instruments. The possibility of cross contamination is also noted in handles for refrigerators, ovens, food dryers, hand washing. It was also noted that there were too much work on the nutritionist and cook, additional personnel need to be added. lack of space, deterioration of facilities were identified in some high school foodservice systems. ATP bioluminescence assay was conducted on surface of food facilities, ATP ranged 1,393±5,041.2 RLU on yellow gloves, 244±258.7 RLU on pink gloves, 3,780±11,418.6 RLU on apron, 49,056±62,831.4 RLU on refrigerator grip, 41,422±61,259.8 RLU in oven, 31,407±41,344.9 RLU on hand cleaning board.

An investigation was conducted to evaluate the hygienic status of 33 high school foodservice systems in Yongin city by using hygiene management guide checklist, ATP bioluminescence assay and microbe inspection petrifilm (APC, coliform group, Staphylococcus aureus) of food utensils during use. The 22 hygiene management guide checklist items about facilities, personal hygiene, food control, distribution, washing and disinfection had good grade but there were some inadequate behaviors on observation. The inspection results showed their sanitary condition met the level B of the recommendation of Korea method, it means sanitary management system get settled but more practical CCP system was needed. ATP bioluminescence assay was conducted on surface of food facilities, ATP ranged 425~2,552 RLU on gloves, 541~70,251 RLU on apron, 1,596~88,490 RLU on working desk, 1,177~263,813 RLU on sterilizer grip, 715~32,814 RLU on sterilizer shelf, 114~619,725 RLU on refrigerator grip, 677~319,007 RLU on refrigerator shelf, 71~196,725 RLU on freezer grip, 1,535~233,375 RLU on freezer shelf. APC ranged 66.7±29.0 CFU on freezer grip, 102.1±35.9 CFU on refrigerator grip, 45.4±28.2 CFU on heating cabinet grip, 58.8±40.4 CFU on sterilizer grip, the number of coliform group ranged 5.6±4.9 CFU on freezer grip, 9.1±8.7 CFU on refrigerator grip, 1.2±1.1 CFU on heating cabinet grip, 4.5±4.4 CFU on sterilizer grip. S. aureus ranged 8.0±5.6, CFU on freezer grip, 12.2±9.6 CFU on refrigerator grip, 2.1±1.6 CFU on heating cabinet grip, 11.6±6.4 CFU on sterilizer grip.

The mitochondrial COI gene has often been utilized as a molecular marker for species identification. However, it has sometimes caused misidentification for some pairs of closely related species. For detecting complementary barcoding loci, we first screened candidate genes by calculating genetic distances within and between species based on 542 sequences collected from the Genbank by using aphids of the Eriosomatini as an example. Of eight genes analyzed, we selected the ATP6 and ATP8 genes, which exhibited lower intraspecific and higher interspecific genetic divergences than did the COI gene. Secondly, we tested the usefulness of these genes by calculating genetic distances between all the combinations of 44 individuals of 23 eriosomatine species for each of the ATP6, ATP8, and COI genes. In the ATP8 gene, the average intraspecific divergence was lowest (0.6%) and the average interspecific divergence was highest (14.7%). The ATP8 gene evolved more rapidly than did the COI gene if genetic divergence between individuals was sufficiently large, whereas it evolved more slowly than did COI if genetic divergence was less than a threshold (1% in COI distance). As a result, species with intraspecific variation in COI and ATP6 exhibited no genetic variation in ATP8. The pattern of genetic divergence in ATP8 well accorded with the pattern of species delimitation in the present taxonomic system. Thus, we conclude that the use of the ATP8 gene in DNA barcoding could improve the accuracy of species identification in the Eriosomatini and possibly other insect groups.

The ubiquitous Na, K-ATPase is a membrane-bound ion pump located in the plasma membrane in all animal cells and plays an essential role in a variety of cellular functions. Studies in several organisms have shown that this protein regulates different aspects of embryonic development and is responsible for the pathogenesis of several human diseases. Na, K-ATPase is an important factor for retinal development, and combinations of the isoforms of each of its subunits are expressed in different cell types and determine its functional properties. In this study, we performed RT-PCR assay to determine temporal expression and in situ hybridization to determine spatial expression of Na, K-ATPase β2 isoform (atp1b2) in Xenopus laevis. Focusing on retinal expression to distinguish the specific expression domain, we used retinal marker genes sox4, sox11, vsx1, and pax6. Xenopus atp1b2 was expressed from late gastrulation to the tadpole stage. Using whole mount in situ hybridization, we showed that Xenopus atp1b2 was expressed broadly in the eye, the whole surface ectoderm, and gills. In situ hybridization on sections revealed detailed and specific expression in the outer nuclear layer of the retina, which consists of two major classes of photoreceptors, rods and cones, surface ectoderm, pharyngeal epithelium, and gills. These findings indicate that atp1b2 may play an important role for the development of Xenopus retina.

Since airborne fungi have been known to aggravate indoor air quality, studies on developing anti-fungal filters increase recently. In this study, silver (Ag) nanoparticle was selected as anti-fungal agent. HEPA filter was coated with silver nanoparticles which were generated via spark discharge system operating at atmospheric pressure and temperature. The anti-fungal effect of the Ag-filter was evaluated with the conventional culture assay. When the number of Ag nano particle per a fungal particle in the filter was 1.91X106, the fungicidal efficiency was higher than 99%. As another anti-fungal test, ATP bioluminiscence detection method was also carried out and the results were correlated with those of the culture assay.

Na+/K+-ATPase, an energy-transducing ion pump, is responsible for maintenance of relatively high concentrations of potassium ions but low concentrations of sodium ions in the cell by transport of these ions across the plasma membrane. Na+/K+-ATPase consists of α, β, and γ subunits, but only α and β subunits are needed for basic functions. Na+/K+-ATPase is also involved in regulation of intracellular calcium ion concentration by coupling with Na+/Ca2+ exchanger involved in intracellular calcium extrusion. Our previous study showed that calcium regulatory molecules including Na+/Ca2+ exchanger are expressed in the uterine endometrium during the estrous cycle and pregnancy in pigs, however, expression of Na+/K+-ATPase in the uterine endometrium has not been determined. Thus, we examined expression of α1 (ATP1A1) and β1 (ATP1- B1) subunits of Na+/K+-ATPase in the uterine endometrium during the estrous cycle and pregnancy in pigs. Real-time RT-PCR analysis showed that levels of ATP1A1 m- RNA in the uterine endometrium during the estrous cycle and early pregnancy were higher than those during mid and term pregnancy, and that levels of ATP1B1 mRNA were highest on day (D) 12 of the estrous cycle. In situ hybridization analysis revealed that ATP1A1 and ATP1B1 mRNAs were localized to luminal (LE) and glandular epithelia (GE) in the endometrium. During mid to term pregnancy, localization of ATP1A1 mRNA was confined to LE, GE, and chorionic membrane (CM) of areolae and ATP1- B1 mRNA was localized to LE, GE and CM with the strongest intensity in LE of areolae. Signal intensity of ATP1B1 mRNA in LE was slightly stronger than that in GE. RT-PCR analysis showed that ATP1A1 and ATP1B1 mRNAs were expressed in conceptuses on D12 and D15 of pregnancy. These results showed that ATP1A1 and ATP1B1 were expressed in the uterine endometrium and conceptuses during the estrous cycle and pregnancy in a pregnancy status- and stage-specific manner. These suggest that Na+/K+-ATPase may play a key role in the establishment and maintenance of pregnancy by regulating intracellular concentrations of various ions including calcium at the maternal-fetal interface in pigs.

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 ± 6.77 RLU/cm²). 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.

Adenosine 5'-triphosphate (ATP) is an important extracellular signaling molecule which is involved in a variety of physiological responses in many different tissues and cell types, by acting at P2 receptors, either ionotropic (P2X) or G protein-coupled metabotropic receptors (P2Y). P2X receptors have seven isoforms designated as P2x₁-P2x7. In this study, we investigated the electrophysiological and pharmacological properties of rat P2x₁-P2x₄currents by using whole-cell patch clamp technique in a heterologous expression system. When ATP-induced currents were analyzed in human embryonic kidney (HEK293) cells following transient transfection of rat P2x₁-P2x₄,the currents showed different pharmacological and electrophysiological properties. ATP evoked inward currents with fast activation and fast desensitization in P2x₁_ or P2x ₃ expressing HEK293 cells, but in P2X₂- or P2x₄- expressing HEK293 cells, ATP evoked inward currents with slow activation and slow desensitization. While PPADS and suramin inhibited P2X₂or P2X₃receptor-mediated currents, they had little effects on P2x₄ receptor-mediated currents. Ivermectin potentiated and prolonged P2x₄ receptor-mediated currents, but did not affect P2X₂ or P2X₃ receptor-mediated currents. We suggest that distinct pharmacological and electrophysiological properties among P2X receptor subtypes would be a useful tool to determine expression patterns of P2X receptors in the nervous system including trigeminal sensory neurons and microglia.

The effects of adenosine 5′-triphosphate (ATP) and ATP analogs, P/sub 2y/ purinoceptor agonists, on growth of normal mouse mammary epithelial cells (NMuMG) were examined. Cells were plated onto 24 well plates in DMEM supplemented with 10 % fetal calf serum. After serum starvation for 24 hours, ATP, P/sub 2y/ purinoceptor agonists (AdoPP[NH]P, ATP-α-S, ATP-γ-S, β, γ-me-ATP and 2me-S-ATP), P/sub 2u/ purinoceptor agonist (UTP) and P/sub 2y/ purinoceptor antagonists (Reactive Blue 2, more selective to P/sub 2y/ receptor than PPADS; PPADS) were added. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA (1 hour pulse with 1 μ Ci/ml, 18～19 hours after treatment). ATP, Adopp[NH]P, ATP-α-S or ATP-γ-S, significantly increased DNA synthesis at 1, 10 and 100 μM concentrations with dose-dependency (P<0.05), and the maximum responses of ATP and ATP analogs were shown at 100 μM concentration (P<0.05). The potency order of DNA synthesis was ATP≥ATP- γ -S>Adopp [NH]P>ATP-α-S. β, γ -me-ATP, 2me-S-ATP and UTP did not increase DNA synthesis. In autoradiographic analysis of percentage of S-phase cells, similar results were observed to those of DNA synthesis. Addition of 1, 10 or 100 μM Reactive Blue 2 or PPADS significantly decreased ATP (100 μM)-induced DNA synthesis, however, PPADS was less effective than Reactive Blue 2. In Elvax 40P implant experiment, ATP directly stimulated mammary endbud growth in situ suggesting the physiological regulator of ATP in mammary growth. ATP 100 μM rapidly increased MAPK activity, reaching a maximum at 5 min and then gradually decreasing to the base level in 30 min. ATP analogs, Adopp[NH]P and ATP-γ-S also increased MAPK activity, however, β, γ-me-ATP and 2me-S-ATP did not. The inhibitor of the upstream MAPK kinase (MEK), PD 98059 (25 μM), effectively reduced ATP (100 μM) or EGF(10 ng/ml, as positive control)-induced MAPK activity and DNA synthesis (P<0.05). These results indicate that ATP-induced DNA synthesis was prevented from the direct inhibition of MAPK kinase pathway. Overall results support the hypothesis that the stimulatory effects of normal mouse mammary epithelial growth by addition of ATP or ATP analogs are mediated through mammary tissue specific P/sub 2y/ purinoceptor subtype, and MAPK activation is necessary for the ATP-induced cell growth.