• Title, Summary, Keyword: ATP

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Electrical properties and ATP-sensitive K+ channel density of the rat substantia nigra pars compacta neurons (랫드 흑질 신경세포의 전기적 특성과 ATP-sensitive K+채널의 전류밀도)

  • Han, Seong-kyu;Park, Jin-bong;Ryu, Pan-dong
    • Korean Journal of Veterinary Research
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    • v.40 no.2
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    • pp.275-282
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    • 2000
  • Substantia nigra is known to highly express glibenclamide binding site, a protein associated to ATP-sensitive $K^{+}$ ($K_{ATP}$) channel in the brain. However, the functional expression of $K_{ATP}$ channels in the area is not yet known. In this work, we attempted to estimate the functional expression of $K_{ATP}$ channels in neurons of the substantia nigra pars compacta (SNC) in young rats using slice patch clamp technique. Membrane properties and whole cell currents attributable to $K_{ATP}$ channel were examined by the current and voltage clamp method, respectively. In SNC, two sub-populations of neurons were identified. Type I (rhythmic) neurons had low frequency rebound action potentials ($4.5{\pm}0.25Hz$, n=75) with rhythmic pattern. Type II (phasic) neurons were characterized by faster firing ($22.7{\pm}3.16Hz$, n=12). Both time constants and membrane capacitance in rhythmic neurons ($34.0{\pm}1.27$ ms, $270.0{\pm}11.83$ pF) and phasic neurons ($23.7{\pm}4.16$ ms, $184{\pm}35.2$ pF) were also significantly different. The current density of $K_{ATP}$ channels was $6.1{\pm}1.47$ pA/pF (2.44~15.43 pA/pF, n=8) at rhythmic neurons of young rats. Our data show that in SNC there are two types of neurons with different electrical properties and the density of $K_{ATP}$, channel of rhythmic neuron is about 600 channels per neuron.

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Protection of Metal Stress in Saccharomyces cerevisiae: Cadmium Tolerance Requies the Presence if Two ATP-Binding Domains of Hsp 104 Protein

  • Lee, Gyeong Hui;Eom, Jeong Hun
    • Bulletin of the Korean Chemical Society
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    • v.22 no.5
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    • pp.514-518
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    • 2001
  • We have explored the importance of two ATP binding domains of Hsp104 protein in protection of yeast cells from cadmium exposure. In the previous study we have discovered that the presence of two ATP binding sites was essential in providing heat sh ock protection as well as rescuing cells from oxidative stress. In this paper we first report wild type cell with functional hsp104 gene is more resistant to cadmium stress than hsp104-deleted mutant cell, judging from decrease in survival rates as a result of cadmium exposure. In order to demonstrate functional role of two ATP binding sites in cadmium defense, we have transformed both wild type (SP1) and hyperactivated ras mutant (IR2.5) strains with several plasmids differing in the presence of ATP binding sites. When an extra copy of functional hsp104 gene with both ATP binding sites was overexpressed with GPD-promoter, cells showed increased survival rate against cadmium stress than mutants with ATP binding sites changed. The degree of protection in the presence of two ATP binding sites was similarly observed in ira2-deleted hyperactivated ras mutant, which was more sensitive to oxidative stress than wild type cell. We have concluded that the greater sensitivity to cadmium stress in the absence of two ATP binding sites is attributed to the higher concentration of reactive oxygen species (ROS) produced by cadmium exposure based on the fluorescence tests. These findings, taken all together, imply that the mechanism by which cadmium put forth toxic effects may be closely associated with the oxidative stress, which is regulated independently of the Ras-cAMP pathway. Our study provides a better understanding of cadmium defense itself and cross-talks between oxidative stress and metal stress, which can be applied to control human diseases due to similar toxic environments.

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Permeability properties of skeletal muscle ATP-sensitive K+ channels reconstituted into planar lipid bilayer (평지방막에 융합된 골격근의 single ATP-sensitive K+ channel의 이온투과성에 대한 연구)

  • Ryu, Pan-dong
    • Korean Journal of Veterinary Research
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    • v.32 no.4
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    • pp.543-553
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    • 1992
  • Properties of unitary ATP-sensitive $K^+$ channels were studied using planar lipid bilayer technique. Vesicles were prepared from bullfrog (Rana catesbeiana) skeletal muscle. ATP-sensitive $K^+$ (K (ATP)) channels were identified by their unitary conductance and sensitivity to ATP. In the symmetrical solution containing 200mM KCI, 10mM Hepes, 1mM EGTA and pH 7.2, single K (ATP) channels showed a linear current-voltage relations with slight inward rectification. Slope conductance at reversal potential was $60.1{\pm}0.43$ pS(n=3)). Micromolar ATP reversibly inhibited the channel activity when applied to the cytoplasmic side. In the range of -50~+50 mV, the channel activity was not voltage-dependent, but the channel gating within a burst was more frequent at negative voltage range. Varying the concentrations of external/internal KCl(mM) to 40/200, 200/200, 200/100 and 200/40 shifted reversal potentials to $-30.8{\pm}2.9$(n=3), $-1.1{\pm}2.7$(n=3), 10.5 and 30.6(mV), respecrivety. These reversal potentials were close to the expected values by the Nernst equation, indicating nearly ideal selectivity for $K^+$ over $Cl^-$. Under bi-ionic conditions of 200mM external test ions and 200mM internal $K^+$, the reversal potentials for each test ion/K pair were measured. The measured reversal potentials were used for the calculation of the releative permeability of alkali cations to $K^+$ ions using the Goldman-Hodgkin-Katz equation. The permeability sequence of 5 cations relative to $K^+$ was $K^+$(1), $Rb^+$(0.49), $Cs^+$(0.27), $Na^+$(0.027) and $Li^+$(0.021). This sequence was recognized as Eisenman's selectivity sequence IV. In addition, modelling the permeation of $K^+$ ion through ATP-sensitive $K^+$ channel revealed that a 3-barrier 2-site multiple occupancy model can reasonably predict the observed current-voltage relations.

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Extracellular ATP Induces Apoptotic Signaling in Human Monocyte Leukemic Cells, HL-60 and F-36P

  • Yoon, Mi-Jung;Lee, Hae-Jin;Kim, Jae-Hwan;Kim, Dong-Ku
    • Archives of Pharmacal Research
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    • v.29 no.11
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    • pp.1032-1041
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    • 2006
  • Extracellular adenosine 5'-triphosphate (ATP) affects the function of many tissues and cells. To confirm the biological activity of ATP on human myeloid leukemic cells, F-36P and HL-60, cells were treated with a variety of concentrations of ATP. The stimulation with extracellular ATP induced the arrest of cell proliferation and cell death. from the analysis of Annexin-V staining and caspase activity by flow cytometry. The Annexin-V positive cells in both cell lines were dramatically increased following ATP stimulation. The expression of P2 purinergic receptor genes was confirmed, such as P2X1, P2X4, P2X5, P2X7 and P2Y1, P2Y2, P2Y4, P2Y5, P2Y6, P2Y11 in both leukemic cell lines. Interestingly, ATP induced intracellular calcium flux in HL-60 cells but not in F-36P cells, as determined by Fluo-3 AM staining. Cell cycle analysis revealed that ATP treatment arrested both F-36P and HL-60 cells at G1/G0. Taken together, these data showed that extracellular ATP via P2 receptor genes was involved in the cell proliferation and survival in human myeloid leukemic cells, HL-60 and F-36P cells by the induction of apoptosis and control of cell cycle. Our data suggest that treatment with extracellular nucleotides may be a novel and powerful therapeutic avenue for myeloid leukemic disease.

Medullary Sponge Kidney Detected in the Pediatric Age (소아 연령에서 발견된 medullary sponge kidney)

  • Chung, Woo Yeong;Cho, Min Hyun;Gu, Young-Ran;Leem, Sun-Hee;Cheong, Hae Il
    • Childhood Kidney Diseases
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    • v.16 no.2
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    • pp.109-114
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    • 2012
  • Purpose: Medullary sponge kidney (MSK) is a rare congenital disease characterized by diffuse ectasia or dilatation of precalyceal collecting tubules. MSK incidence and prevalence in the general population is uncertain and only a few patients are reported especially in the pediatric age. There has been increasing reports of patients with MSK who have other malformative disorders. Also several case reports concerning about etiological association of some genes. Methods: Collaborative study through nation-wide survey was done to investigate the incidence and etiological association of some genes such as GDNF gene, ATP6V1B1, ATP6V0A4 gene in developing MSK in Korean children. Results: Four cases of MSK who have various other malformative disorders were collected. There are no mutations of GDNF gene, ATP6V1B1, ATP6V0A4 gene in all patients. Conclusion: MSK is one of the very rare diseases in pediatric age. The etiological association of GDNF gene, ATP6V1B1, ATP6V0A4 gene in developing MSK in Korean children is not proved.

The Signaling Mechanism of Contraction Induced by ATP and UTP in Feline Esophageal Smooth Muscle Cells

  • Kwon, Tae Hoon;Jung, Hyunwoo;Cho, Eun Jeong;Jeong, Ji Hoon;Sohn, Uy Dong
    • Molecules and Cells
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    • v.38 no.7
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    • pp.616-623
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    • 2015
  • P2 receptors are membrane-bound receptors for extracellular nucleotides such as ATP and UTP. P2 receptors have been classified as ligand-gated ion channels or P2X receptors and G protein-coupled P2Y receptors. Recently, purinergic signaling has begun to attract attention as a potential therapeutic target for a variety of diseases especially associated with gastroenterology. This study determined the ATP and UTP-induced receptor signaling mechanism in feline esophageal contraction. Contraction of dispersed feline esophageal smooth muscle cells was measured by scanning micrometry. Phosphorylation of $MLC_{20}$ was determined by western blot analysis. ATP and UTP elicited maximum esophageal contraction at 30 s and $10{\mu}M$ concentration. Contraction of dispersed cells treated with $10{\mu}M$ ATP was inhibited by nifedipine. However, contraction induced by $0.1{\mu}M$ ATP, $0.1{\mu}M$ UTP and $10{\mu}M$ UTP was decreased by U73122, chelerythrine, ML-9, PTX and $GDP{\beta}S$. Contraction induced by $0.1{\mu}M$ ATP and UTP was inhibited by $G{\alpha}i_3$ or $G{\alpha}q$ antibodies and by $PLC{\beta}_1$ or $PLC{\beta}_3$ antibodies. Phosphorylated $MLC_{20}$ was increased by ATP and UTP treatment. In conclusion, esophageal contraction induced by ATP and UTP was preferentially mediated by P2Y receptors coupled to $G{\alpha}i_3$ and $G{\alpha}q$ proteins, which activate $PLC{\beta}_1$ and $PLC{\beta}_3$. Subsequently, increased intracellular $Ca^{2+}$ and activated PKC triggered stimulation of MLC kinase and inhibition of MLC phosphatase. Finally, increased $pMLC_{20}$ generated esophageal contraction.

효소적 방법에 의한 참돔(Pagrus major) 품질판정에 관한 연구

  • 배진한;정호진;여해경;심길보;김태진;조영제
    • Proceedings of the Korean Aquaculture Society Conference
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    • pp.153-154
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    • 2003
  • 운동여부와 운동횟수 그리고 어체 크기에 대한 양식산 어종의 품질판정을 위한 지표를 검색하기 위하여 효소적 방법 즉 ATP 관련화합물과 양식산 활어의 건강한 상태와 칼슘의 노출, 염도의 감소, 산소의 부족, 궁핍 등과 같은 스트레스를 받은 상태를 판단하는 adenylate energy charge (AEC)를 근육의 품질판정을 위한 지표로써 조사하였다. 운동사육구와 비운동사육구에서의 참돔육에 대한 ATP 관련화합물의 변화는 운동 0일에는 ATP함량은 8.62 μmol/g이었지만 20일차에는 8.59 μmol/g으로 운동기간이 길어짐에 따라 ATP의 함량은 약간의 감소를 보였다. 이는 운동이라는 자극요인으로 인하여 ATP의 소모가 촉진되었을 것으로 판단된다. 또한 운동 0일차의 ADP함량은 0.72 μmo1/g이었지만 20일차에는 0.93μmol/g로 ATP의 소모만큼 약간의 증가를 보이고 있다. 반면, 비운동사육구에서 0일차의 ATP함량은 5.78 μmol/g이었지만 20일차에서는 6.31 μmol/g로 운동사육구와 대조적으로 기간이 길어짐에 따라 ATP함량이 약간의 증가를 보여주고 있다. 생리적으로 동물에 가혹한 환경에 두거나 스트레스를 가하면 adenylate energy charge (AEC)가 영향을 받아 변화게 된다. AEC는 (ATP + ½ ADP)/(ATP + ADP + AMP)의 비율로 정의되는데, AEC의 범위는 0∼1사이로써 0은 ATP 관련화합물이 AMP로 전환된 상태를 말하며, 1은 모든 ATP 관련 화합물이 ATP로 존재하는 것을 의미한다. 그러므로, 이들결합의 상관적인 정도는 개개의 세포에서 에너지의 효용성을 측정하는 것으로 이용되고 있다. AEC 수치는 운동수조에서 0.91∼0.92 건강상태가 좋았지만 비운동수조는 0.87∼0.88로 운동수조에 비해 값이 낮게 나왔다. 운동횟수에 관계없이 운동시킨 어육의 AEC는 0.90∼0.91로 건강한 상태였다. 어체크기에 따른 ATP 관련화합물의 변화는 0.7∼0.9kg의 크기를 가진 참돔에서는 ATP 함량이 7.88 μmole/g, 1.5∼l,7kg의 크기를 가진 참돔에서는 ATP함량이 9.61μmole/g로 많았고 다른 크기에서는 큰 차이를 나타내지 않았다. 그리고 ADP함량은 어체의 크기가 증가할수록 함량은 감소하였다. 그러나, 어체의 크기에 따른 ATP관련화합물의 변화는 거의 없었다. 건강도의 지표인 AEC는 0.89∼0.94로 전체적으로 건강상태로 나타났다. 하루 2회 운동시킨 육에서의 ATP함량은 0일차에 8.75 μmole/g, 10일차에 8.46 μmole/g, 20일차에 8.51μmole/g로 약간의 감소를 보여준다. ADP함량은 일까지는 증가하지만 그 이후에는 큰 변화를 보이지 않는다. 6시간 운동시킨 그룹의 ATP함량은 0일차에 8.63 μmole/g, 5일차에 8.52 μmole/g, 10일차에 8.4μmole/g, 20일차에 8.51 μmole/g로 10일까지는 감소하지만 그 이후에는 큰 변화를 보이지 않는다. ADP함량은 0일차에 0.72 μmole/g, 10일차 0.84 μmole/g, 20일차에 0.83 μmole/g으로 10일차까지는 약간 증가하지만 그 이후에는 큰 변화를 보이지 않았다. 참돔의 근원섬유에 있어서 Ca/sup 2+/이 존재할 때 Mg/sup 2+/-ATPase 활성의 변화는 운동시킨 참돔 근원섬유의 Mg/sup 2+/-ATPase 활성은 0.35μmol Pi/minㆍmg였으며, 비운동구에서 0.34μmol Pi/minㆍmg으로 운동시킨 참돔의 육에서 ATPase 활성이 약간 높은 것으로 나타났다. 운동사육시킨 참돔은 운동기간이 증가할수록 ATPase활성이 증가하여 운동 20일에는 0.42μmo1 Pi/minㆍmg의 함량을 나타내었으나, 비운동사육에서는 큰 차이를 보이지 않고 있다. 근원섬유의 Ca/sup 2+/가 첨가되지 않았을때의 Mg/sup 2+/-ATPase 활성을 나타내었다. 운동시키전에는 각각 0.10μmol Pi/minㆍmg, 0.14μmol Pi/minㆍmg 나타나 비운동구가 운동구에 비하여 Mg/sup 2+/(-Ca/sup 2+/)-ATPase 활성이 높았으며 사육기간동안 활성이 서서히 감소하는 경향을 나타내었다. Ca/sup 2+/-ATPase의 활성 또한 사육기간 동안 큰 변화를 보이지 않았다. 그러므로, 참돔의 운동여부에 따라서 ATPase활성을 살펴본 결과, 운동기간에 따른 약간의 차이는 보이고 있으나, 품질지표로써 사용하기가 어렵다고 판단된다. 효소적인 방법을 이용한 양식산 활어의 품질지표로써 ATP 관련화합물, ATPase 활성은 운동여부, 크기에 따라 객관적인 지표로써의 사용이 어려우므로, 건강도를 측정할 수 있는 AEC의 수치를 적용하는 것이 적합하다고 판단된다. 즉, 양식산 참돔의 AEC 수치가 1∼0.9의 범위에서는 건강한 활어로 상급의 등급을, 0.8∼0.7의 AEC 수치는 중급의 등급으로, 그 이하의 수치에서는 하급으로 적용가능 하다고 판단된다.

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Development of Intelligent ATP System Using Genetic Algorithm (유전 알고리듬을 적용한 지능형 ATP 시스템 개발)

  • Kim, Tai-Young
    • Journal of Intelligence and Information Systems
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    • v.16 no.4
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    • pp.131-145
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    • 2010
  • The framework for making a coordinated decision for large-scale facilities has become an important issue in supply chain(SC) management research. The competitive business environment requires companies to continuously search for the ways to achieve high efficiency and lower operational costs. In the areas of production/distribution planning, many researchers and practitioners have developedand evaluated the deterministic models to coordinate important and interrelated logistic decisions such as capacity management, inventory allocation, and vehicle routing. They initially have investigated the various process of SC separately and later become more interested in such problems encompassing the whole SC system. The accurate quotation of ATP(Available-To-Promise) plays a very important role in enhancing customer satisfaction and fill rate maximization. The complexity for intelligent manufacturing system, which includes all the linkages among procurement, production, and distribution, makes the accurate quotation of ATP be a quite difficult job. In addition to, many researchers assumed ATP model with integer time. However, in industry practices, integer times are very rare and the model developed using integer times is therefore approximating the real system. Various alternative models for an ATP system with time lags have been developed and evaluated. In most cases, these models have assumed that the time lags are integer multiples of a unit time grid. However, integer time lags are very rare in practices, and therefore models developed using integer time lags only approximate real systems. The differences occurring by this approximation frequently result in significant accuracy degradations. To introduce the ATP model with time lags, we first introduce the dynamic production function. Hackman and Leachman's dynamic production function in initiated research directly related to the topic of this paper. They propose a modeling framework for a system with non-integer time lags and show how to apply the framework to a variety of systems including continues time series, manufacturing resource planning and critical path method. Their formulation requires no additional variables or constraints and is capable of representing real world systems more accurately. Previously, to cope with non-integer time lags, they usually model a concerned system either by rounding lags to the nearest integers or by subdividing the time grid to make the lags become integer multiples of the grid. But each approach has a critical weakness: the first approach underestimates, potentially leading to infeasibilities or overestimates lead times, potentially resulting in excessive work-inprocesses. The second approach drastically inflates the problem size. We consider an optimized ATP system with non-integer time lag in supply chain management. We focus on a worldwide headquarter, distribution centers, and manufacturing facilities are globally networked. We develop a mixed integer programming(MIP) model for ATP process, which has the definition of required data flow. The illustrative ATP module shows the proposed system is largely affected inSCM. The system we are concerned is composed of a multiple production facility with multiple products, multiple distribution centers and multiple customers. For the system, we consider an ATP scheduling and capacity allocationproblem. In this study, we proposed the model for the ATP system in SCM using the dynamic production function considering the non-integer time lags. The model is developed under the framework suitable for the non-integer lags and, therefore, is more accurate than the models we usually encounter. We developed intelligent ATP System for this model using genetic algorithm. We focus on a capacitated production planning and capacity allocation problem, develop a mixed integer programming model, and propose an efficient heuristic procedure using an evolutionary system to solve it efficiently. This method makes it possible for the population to reach the approximate solution easily. Moreover, we designed and utilized a representation scheme that allows the proposed models to represent real variables. The proposed regeneration procedures, which evaluate each infeasible chromosome, makes the solutions converge to the optimum quickly.

Indentification of IMP in the Muscle of the Ascidian Halocynthia roretzi and Changes of ATP Breakdown Products during Storage (우렁쉥이 Halocynthia roretzi 근육중 IMP의 분포확인 및 저장중 ATP분해생성물의 변화)

  • PARK Choon-Kyu;KIM Woo-Jun;KANG Hoon-I;KANG Tae-Jung;SHIN Suk-Woo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.2
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    • pp.140-148
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    • 1994
  • Indentification of IMP was carried out and changes in ATP breakdown products during storage at $0^{\circ}C\;ad\;20^{\circ}C$ were investigated in the muscles of ascidian Halocynthia roretzi. For identifying IMP, the ion-exchange column chromatographic method was applied to the perchloric acid extract of the muscle of cultured ascidian collected at the southern coast near Chungmu of Korea in April 1989. The IMP of sample was eluted a little earlier than that of the reference standard, but absorption spectra of both fractions agreed each other. In addition, both fractions gave the identical retention time of HPLC. These results reconfirmed that the ascidian muscle did contain IMP, indicating that ATP was degraded through IMP breakdown pathway, such as $ATP{\to}ADP{\to}AMP{\to}IMP{\to}Ino{\to}Hyp$. Ado was detected in some samples and IMP was detected throughout the experimental periods at both temperatures, but their levels were always very low; they did not increase significantly even when the decreasing rate of AMP was very rapid and concomitant remarkable increase in Ino were observed at the early stage of storage. Those changes in ATP suggest that AMP deaminase activity was present in the ascidian muscle, though it was very low. The main breakdwon pathway of ATP was assumed to be $ATP{\to}ADP{\to}AMP{\to}Ado{\to}Ino{\to}Hyp$. In conclusion, there were two breakdown pathways of ATP in the muscle of ascidian as was the case for the muscle of many marine crustaceans.

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Safety and Feasibility of Thallium-201 Myocardial SPECT with Intravenous Infusion of Disodium Adenosine Triphosphate (ATP) in the Diagnosis of Coronary Artery disease (관상동맥 질환 진단에서 ATP부하 T1-201 심근 관류 SPECT의 안전성과 유용성)

  • Pai, Moon-Sun;Park, Chan-H.;Yoon, Seok-Nam;Kim, Won;Kim, Han-Soo
    • The Korean Journal of Nuclear Medicine
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    • v.32 no.3
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    • pp.250-258
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    • 1998
  • Purpose: ATP (adenosine triphosphate) is a potent coronary vasodilator with a rapid onset of action and a very short half-life. Myocardial perfusion scintigraphy with intravenous ATP has not yet been sufficiently proven in the diagnosis, follow-up, and risk stratification of coronary artery disease. The purpose of this study was to evaluate the safety, feasibility and diagnostic accuracy of pharmacologic stress thallium-201 myocardial SPECT using an intra-venous ATP infusion in patients with suspected coronary artery disease. Materials and Methods: Thallium-201 myocardial SPECT in 319 patients with suspected coronary artery disease were performed after the infusion of ATP (0.08 mg/kg/min for 6 min). The adverse effects were carefully monitored. Coronary angiography was also performed within 3 weeks. Results: Although 76.5% of the patients had some adverse effects, they were transient, mild, and well tolerated. In all patients, the ATP infusion protocol was completed and only 2 patients required aminophylline. The adverse effects were dyspnea in 63%, headache in 31%, flushing in 21%, chest pain in 14% and abdominal discomfort in 5% of the patients. The sensitivity and specificity were 80% and 90% respectively. Conclusion: Thallium-201 myocardial SPECT after 6 min-infusion of ATP at a rate of 0.08 mg/kg/min is safe and has a diagnostic value in detecting coronary artery disease.

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