• 한국식품위생안전성학회지
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• 제15권3호
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• pp.252-255
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• 2000

본 연구는 육류와 가금육, 유가공 공장의 생산라인에서 미생물 오염수준을 측정함에 있어 ATP bioluminescence법이 적용될 수 있는지를 조사하기 위해 수행되었다. 도축장, 도계장, 유가공 공장 생산라인에서 시료를 채취하였으며 ATP bioluminescence법과 표준평판법을 병행하여 나타난 결과치(log RLU/ml, CFU(log/ml))를 상호 비교하여 상관계수를 측정하였다. 모든 시료(n=408)에서의 상관계수는 0.93이었으며 이중 쇠고기, 돼지고기, 닭고기는 0.93(n=220), 육, 유가공장 생산라인 또한 0.93(n=187)로 비교적 높게 나타나 ATP bioluminescence법이 기존의 표준평판법에 비해 상관계수가 높으므로 식품생산현장에서 신속하고 편리하게 세균의 오염여부를 판정할 수 있는 방법이라고 생각된다.

• The Korean Journal of Physiology and Pharmacology
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• 제13권3호
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• pp.175-179
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• 2009

High concentrations of ATP induce membrane blebbing. However, the underlying mechanism involved in epithelial cells remains unclear. In this study, we investigated the role of the P2X7 receptor (P2X7R) in membrane blebbing using Par C5 cells. We stimulated the cells with 5 mM of ATP for 1${\sim}$2 hrs and found the characteristics of membrane blebbing, a hallmark of apoptotic cell death. In addition, 500 ${\mu}M$ Bz-ATP, a specific P2X7R agonist, induced membrane blebbing. However, 300 ${\mu}M$ of Ox-ATP, a P2X7R antagonist, inhibited ATP-induced membrane blebbing, suggesting that ATP-induced membrane blebbing is mediated by P2X7R. We found that ATP-induced membrane blebbing was mediated by ROCK I activation and MLC phosphorylation, but not by caspase-3. Five mM of ATP evoked a biphasic $[Ca^{2+}]_i$ response; a transient $[Ca^{2+}]_i$ peak and sustained $[Ca^{2+}]_i$ increase secondary to ATP-stimulated $Ca^{2+}$ influx. These results suggest that P2X7R plays a role in membrane blebbing of the salivary gland epithelial cells.

• 한국식품과학회지
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• 제18권2호
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• pp.88-92
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• 1986

Bioluminescence방법에 의하여 세균내에 존재하고 있는 ATP를 추출하여 그양을 정량분석하므로서 제육표면에 생존하고 있는 세균수를 신속하게 측정하는데 응용될 수 있는 가능성을 조사하기 위하여 본 실험을 실시하였으며, 그 결과는 1. Luciferin-luciferase에 작용하는 ATP농도가 $10^{-10}M$부터 $10^{-6}M$까지 증가함에 따라 발생된 빛의 양도 비례적으로 증가했다. 2 . 계육에서 분리동정된 부패균인 E. coli, H, alvei, P. putida, A. hydrophila에서 추출된 ATP 농도와 세균수와의 관계는 거의 일직선으로 나타난다. 3. E. coli에 있는 ATP양은 생존중 거의 변화하지 않는 것으로 나타났다. 4 계육의 저장기간동안 증식된 세균수에 따라 ATP 농도도 비례 적으로 증가되며 이때 상판계수(r)는 0.95로 나타났다. 5. 계육에 오염된 세균수와 ATP 농도와의 관계에서 볼 때 ATP농도가 $10^{-7}M$부근에서부터 부패가 시작된다고 사료된다. 6. 개발된 방법에 의하여 계육표면에 존재하는 세균의 ATP 농도를 측정하여 세균수를 간접적으로 신속히 ( 1시간내) 조사할 수 있다.

• IMMUNE NETWORK
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• 제15권1호
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• pp.44-49
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• 2015

Interactions between microbes and epithelial cells in the gastrointestinal tract are closely associated with regulation of intestinal mucosal immune responses. Recent studies have highlighted the modulation of mucosal immunity by microbe-derived molecules such as ATP and short-chain fatty acids. In this study, we undertook to characterize the expression of the ATP-gated $P2X_7$ receptor ($P2X_7R$) on M cells and its role in gastrointestinal mucosal immune regulation because it was poorly characterized in Peyer's patches, although purinergic signaling via $P2X_7R$ and luminal ATP have been considered to play an important role in the gastrointestinal tract. Here, we present the first report on the expression of $P2X_7R$ on M cells and characterize the role of $P2X_7R$ in immune enhancement by ATP or LL-37.

• The Korean Journal of Physiology and Pharmacology
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• 제8권2호
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• pp.95-100
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• 2004

Ischemic preconditioning (IPC) has been accepted as a heart protection phenomenon against ischemia and reperfusion (I/R) injury. The activation of ATP-sensitive potassium $(K_{ATP})$ channels and the release of myocardial nitric oxide (NO) induced by IPC were demonstrated as the triggers or mediators of IPC. A common action mechanism of NO is a direct or indirect increase in tissue cGMP content. Furthermore, cGMP has also been shown to contribute cardiac protective effect to reduce heart I/R-induced infarction. The present investigation tested the hypothesis that $K_{ATP}$ channels attenuate DNA strand breaks and oxidative damage in an in vitro model of I/R utilizing rat ventricular myocytes. We estimated DNA strand breaks and oxidative damage by mean of single cell gel electrophoresis with endonuclease III cutting sites (comet assay). In the I/R model, the level of DNA damage increased massively. Preconditioning with a single 5-min anoxia, diazoxide $(100\;{\mu}M)$, SNAP $(300\;{\mu}M)$ and 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate (8-pCPT-cGMP) $(100\;{\mu}M)$ followed by 15 min reoxygenation reduced DNA damage level against subsequent 30 min anoxia and 60 min reoxygenation. These protective effects were blocked by the concomitant presence of glibenclamide $(50\;{\mu}M)$, 5-hydroxydecanoate (5-HD) $(100\;{\mu}M)$ and 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate, Rp-isomer (Rp-8-pCPT-cGMP) $(100\;{\mu}M)$. These results suggest that NO-cGMP-protein kinase G (PKG) pathway contributes to cardioprotective effect of $K_{ATP}$ channels in rat ventricular myocytes.

• The Korean Journal of Pain
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• 제18권2호
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• pp.107-112
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• 2005

Background: Nerve ligation injury may produce mechanical allodynia, but this can be reversed after an intrathecal administration of adenosine analogues. In many animal and human studies, ATP-sensitive potassium channel blockers have been known to reverse the antinociceptive effect of various drugs. This study was performed to evaluate the mechanical antiallodynic effects of spinal R-PIA (Adenosine A1 receptor agonist) and the reversal of these effects due to pretreatment with glibenclamide (ATP-sensitive potassium channel blocker). Thus, the relationship between the antiallodynic effects of R-PIA and ATP-sensitive potassium channel were investigated in a neuropathic model. Methods: Male Sprague Dawley rats were prepared by tightly ligating the left lumbar 5th and 6th spinal nerves and implantation of a chronic lumbar intrathecal catheter for drug administration. The mechanical allodynia was measured by applying von Frey filaments ipsilateral to the lesioned hind paw. And the thresholds for paw withdrawal assessed. In study 1, either R-PIA (0.5, 1 and $2{\mu}g$) or saline were administered intrathecally for the examination of the antiallodynic effect of R-PIA. In study 2, glibenclamide (2, 5, 10 and 20 nM) was administered intrathecally 5 min prior to an R-PIA injection for investigation of the reversal of the antiallodynic effects of R-PIA. Results: The antiallodynic effect of R-PIA was produced in a dose dependent manner. In study 1, the paw withdrawal threshold was significantly increased with $2{\mu}g$ R-PIA (P < 0.05). In study 2, the paw withdrawal threshold with $2{\mu}g$ R-PIA was significantly decreased almost dose dependently by intrathecal pretreatment of 5, 10 and 20 nM glibenclamide (P < 0.05). Conclusions: These results demonstrated that an intrathecal injection of ATP-sensitive potassium channel blockers prior to an intrathecal injection of adenosine A1 receptors agonist had an antagonistic effect on R-PIA induced antiallodynia. The results suggest that the mechanism of mechanical antiallodynia, as induced by an intrathecal injection of R-PIA, may involve the ATP-sensitive potassium channel at both the spinal and supraspinal level in a rat nerve ligation injury model.

• International Journal of Oral Biology
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• 제31권4호
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• pp.141-148
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• 2006

The effects of adenosine triphosphate(ATP) on salivary glands have been recognized since 1982. The presence of purinergic recepetors(P2Rs) that mediate the effects of ATP in various tissues, including parotid and submandibular salivary gland, has been supported by the cloning of receptor cDNAs and the expression of the receptor proteins. P2Rs have many subtypes, and the activation of these receptor subtypes increase intracellular $Ca^{2+}$, a key ion in the regulation of the secretion in the salivary gland. The apical pores of taste buds in circumvallate and foliate papillae are surrounded by the saliva from von Ebner salivary gland(vEG). Thus, it is important how the secretion of vEG is controlled. This study was designed to elucidate the roles of P2Rs on salivary secretion of vEG. Male Sprague-Dawley rats (about 200 g) were used for this experiment. vEG-rich tissues were obtained from dissecting $500-1,000\;{\mu}m$ thick posterior tongue slices under stereomicroscope view. P2Rs mRNA in vEG acinar cells were identified with RT-PCR. To observe the change in intracellular $Ca^{2+}$ activity, we employed $Ca^{2+}-ion$ specific fluorescence analysis with fura-2. Single acinar cells and cell clusters were isolated by a sequential trypsin/collagenase treatment and were loaded with $10\;{\mu}M$ fura -2 AM for 60 minutes at room temperature. Several agonists and antagonists were used to test a receptor specificity. RT-PCR revealed that the mRNAs of $P2X_4$, $P2Y_1$, $P2Y_2$ and $P2Y_3$ are expressed in vEG acinar cells. The intracellular calcium activity was increased in response to $10\;{\mu}M$ ATP, a P2Rs agonist, and 2-MeSATP, a $P2Y_1$ and $P2Y_2R$ agonist. However, $300\;{\mu}M\;{\alpha}{\beta}-MeATP$, a $P2X_1$ and $P2X_3R$ agonist, did not elicit the response. The responses elicited by $10\;{\mu}M$ ATP and UTP, a $P2Y_2R$ agonists, were maintained when extracellular calcium was removed. $10\;{\mu}M$ suramin, a P2XR antagonist, and reactive blue 2, a P2YR antagonist, partially blocked ATP-induced response. However, when extracellular calciums were removed, suramin did not abolish the responses elicited by ATP. These results suggest that P2Rs play an important role in salivary secretion of vEG acinar cells and the effects of ATP on vEG salivary secretion may be mediated by $P2X_4$, $P2Y_1$, $P2Y_2$, and/or $P2Y_3$.

• 한국식품영양과학회지
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• 제39권11호
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• pp.1719-1723
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• 2010

창원시 소재 단체급식소 4개소에서 사용하는 칼($12\;cm^2$), 도마($100\;cm^2$), 고무장갑(손바닥과 손가락 사이)의 표면을 대상으로 ATP Luminometer와 일반세균수용 건조필름을 이용한 미생물 배양법을 함께 적용하여 ATP(RLU)와 APC (CFU)을 측정하였다. 통계분석 결과, 3가지 조리도구 모두에서 ATP(RLU)와 APC(CFU) 간에 유의적인 양의 상관성이 존재하였다. 각 도구별 상관계수는 칼 0.84(p<0.001), 도마 0.79(p<0.001), 고무장갑 0.78(p<0.001)로 나타났다. 본 실은 단체급식 현장에 적합한 ATP Luminometer 가이드라인을 설정하기에 앞서 실시간적인 표면 위생 모니터링 도구로서의 사용 적합성을 살펴 본 예비실험이며, 도출된 회귀식을 이용하여 도구별 ATP(RLU) 가이드라인을 설정하고 검증할 예정이다.

• Parasites, Hosts and Diseases
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• 제27권3호
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• pp.161-170
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• 1989

Toxeplasma의 추출액을 3H-casein을 기질로 반응시켰을 때, pH 6.0과 PH 8.5에서 casein을 분해하였으며, pH 6.0에서는 cysteinyl protease의 억제제 인 iodoacetamide(rAh)에 의해 억제되 었고, 활성제 인 dithiothreitol (DTT)에 의해 환성이 증가하였다. 또 pH 8.5에서는 serine protease의 억제제인 phenylmethylsulfonil fluoride (PMSF)에 의해 활성이 억제되었으며, ATP를 첨가할 때 그 활성이 증가하여 ATP 의존성 효소임을 알 수 있었다. 위의 단백질 분해 효소를 부분 정제하기 위해 여러 chromatography를 실시하였는데, 먼저 DE52 (2.Sfx40 cm)에 통과시켰을 때, 0.05M-0.IM NaCl에 의해 유출되는 분획이 pH 6.0에서 황성을 나타내었으며, 0.25V- 0.3M에서 유출되는 분획이 pH 8.5에서 황성을 나타내었다. 이 분회들을 각각 Sephadex G-200 ($2.50{\phi}{\times}40cm$) 에 통과시켜 pH 6.0에서 활성을 나타내는 분획은 exclusion limit내에서, pH 8.5의 분획은 exclusion limit 외에서 분획을 얻었다. 이들을 각각 hydroxylapatite ($2.50{\phi}{\times}10cm$과 $2.5{\phi}{\times}20cm$)를 통과시켜 각각을 0.05M Phosphate로 유출되는 분회에서 높은 환성을 얻었다. 부분 정제된 분획들의 특성을 검토하기 위하여 억제제를 농도별로 처리하였을 때, pH 0.0에서의 분해 효소는 10-3M IAA에 의해 활성이 반감되어 cysteinyl acid protease임을 알 수 있었다. pH 8.5에서의 분해 효소는 10-5M PMSF에 의해 활성이 반감되었고, ATP에 의해 활성이 증가(ATP의 농도가 2.0mM 이상에서는 억제)하여 ATP-dependent neutral serine protease임을 알 수 있었다.

• 미생물학회지
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• 제30권6호
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• pp.528-532
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• 1992

계면 활성제 특히 Triton X-100 의 첨가에 의해 ATP-의존성 효소의 subunit 인 Clp P 의 안정도를 높히고, 염의 첨가에 의한 효소 활성의 유지 정도와 Clp P 호소에 대한 생화학적 성질을 조사하였다. Clp A 가 가지고 있는 ATPase 활성을 나타내기 위하여서는 Clp P 의 단백질 분해 효소활성이 요구되며 Clp 효소의 다량체형성을 조사한 결과 ATP 가 존재하지 않을 경우에는 Clp A 와 Clp P 효소가 각각 용출하였으며 ATP 및 Mg 를 첨가하였을 때에는 complex 를 형성하여 분자량이 600,000 이상으로서 Clp P 효소는 Hexamer 로서 효소활성을 나타내며 Clp A 와 Clp P 의 다량체 형성이 요구되고 형성시에 ATP 가 관여하고 있는 것으로 나타났다.