The content of ATP, ADP, AMP, sodium phosphate and sodium pyrophosphate were determined by 31P NMR, the linear range of ATP, ADP and AMP were found to be 0.004-0.080 mol x L(-1), sodium phosphate and sodium pyrophosphate were 0.005-0.100 mol x L(-1). The RSD were 0.40%-1.30%, the recovery were 96.9% - 105.2%. The method has been applied to the determination of ATP injection. The impurities of ATP injection were ADP and sodium phosphate. The content of ATP is in line with the requirement of the pharmacopoeia. The results indicated that the method is of good reproducibility, high accuracy, rapid and simple operation, without pretreatment and interference of other elements, 31P NMR is a new and reliable method of analyzing ATP, ADP, AMP and phosphate.
Adenosine Diphosphate ; analysis ; Adenosine Monophosphate ; analysis ; Adenosine Triphosphate ; analysis ; Chemistry, Pharmaceutical ; methods ; Diphosphates ; analysis ; Magnetic Resonance Spectroscopy ; Perfusion ; Pharmaceutical Preparations ; analysis ; Phosphates ; analysis ; Quality Control ; Reproducibility of Results

BACKGROUND: To evaluate the metabolic and pathological changes associated with myocardial ischemia-reperfusion, magnetic resonance spectroscopy with 31P was applied as well as pathological examination. METHODS: Effect of adenosine on the recovery of high energy phosphorous metabolites during the reperfusion period following 90 minutes of left anterior descending coronary artery(LAD) ligation was assessed by 31P spectroscopy in 13 cats(8 : control group, 5 : adenosine group). In adenosine group 0.2 mg/kg/min of adenosine was infused intravenously for 90 minutes from 30 minutes before reperfusion. The experiments were peformed on a 4.7 T/30cm Biospec MRS/MRI system(Bruker, Switzerland) and the MR signals were obtained by using innerdiameter 1.5 cm sized doubly tuned surface coil. The size of the spectral peaks was measured by area integration method. RESULTS: In control group, high energy phosphorous metabolites decreased continueously during the ischemic period revealing the lowest values at the end of the periods : 17.0% for PCr and 24.0% for ATP, PCr depleted below 50% of the baseline level immediately after the LAD ligation and ATP, after 15 minutes of ischemia. Therfore the depletion rate was faster in PCr change than in ATP. The recovery of the PCr and ATP occurred after reestablishment of blood flow showing, for example, 43.3% and 36.3% of the baseline levels after 10 minutes of reperfusion. After infusion of adenosine, there was a tendency of higher recovery rates of high energy phosphates than in control group. Recovery rates of PCr and ATP after 90 minutes of reperfusion, were 28.2%, 11.2% in control group and 38.3%, 18.6% in adnosine group, respectively. In adenosine grop, relative sizes of infarction were not statistically different from those of control group. CONCLUSIONS: 31P MRS can be used for in-vivo assessment of the changes of high energy phosphorous metablites concerning acute myocardial ischemia and reperfusion. Adenosine infusion improves the recovery of ATP and PCr during the reperfusion period following acute ischemia.
Adenosine Triphosphate ; Adenosine* ; Animals ; Cats* ; Infarction ; Ischemia ; Ligation ; Magnetic Resonance Spectroscopy ; Myocardial Ischemia ; Phosphates ; Polymerase Chain Reaction ; Reperfusion ; Spectrum Analysis

OBJECTIVETo explore the changes of brain energy metabolism following acrylamide (ACR) poisoning.

METHODSCreatie kinase (CK), adenosine triphosphate (ATP), adenosine diphosphate(ADP), adenosine 5'-monophosphate(AMP) and glucose contents in brain were observed in O1a mice and 6J mice following ACR intoxication by enzyme analytical method.

RESULTSATP, CK and glucose levels decreased transiently in O1a mice, while ATP level in 6J mice was significantly decreased (1.76 mumol/g, P < 0.01), as compared to the control (2.53 mumol/g) but ADP and AMP were increased, glucose was decreased. The activity of CK in poisoned group (1.13 mumol/g, P < 0.01) was lower than that of control (3.16 mumol/g and lasted for 5 weeks).

CONCLUSIONThe influence of ACR on O1a mice was slight and reversible but on 6J mice was severe and lasting. There was severe damage to the potential energy supply compensation, which might be the biochemical basis of neuron damage induced by acrylamide.

Acrylamide ; poisoning ; Adenosine Triphosphate ; analysis ; Animals ; Brain ; drug effects ; metabolism ; Creatine Kinase ; analysis ; Energy Metabolism ; drug effects ; Glucose ; analysis ; Mice

PURPOSE: To evaluate the various phosphorus metabolism' of breast tumors with use of in vivo phosphorus-31 (31P) M R spectroscopy (MRS) MATERIALS AND METHODS: Five patients with breast tumor (benign in two, malignant in three) and three normal healthy volunteers participated in this study. All in vivo31P MRS examinations were performed on 1.5 Twhole-body MRI/MRS system by using a Free Induction Decay (FID) pulse sequence. Tl-weighted MR images were used for localization of tumors. Peak areas for each phosphorus metabolite were measured using a Marquart algorithm. RESULTS: Breast carcinoma had a substantially larger phosphomonoester (PME) and a smaller phosphocreatine (PCr) peak intensity than normal breast tissue. This was reflected in the relatively higher PME/PCr ratio of breast carcinomas as well as phosphodiester (PDE)/PCr, inorganic phosphate (Pi)/PCr, and adenosine triphosphate (ATP)/PCr ratios, compared with normal controls. The mean pH value of breast tumor demonstrating the alkaline nature was higher than that of normal controls. Spectral patterns between benign breast disease and normal breast tissue were quite similar, and differentiation was not established. CONCLUSION: Our preliminary study suggests that in vivo 31P MRS is a noninvasive examination which may be useful in the early differentiation of malignant breast tumors from normal and benign conditions. However, normal control and benign conditions could not be characterized on the basis of the phosphorus metabolite ratios.
Adenosine Triphosphate ; Breast Diseases ; Breast Neoplasms* ; Breast* ; Healthy Volunteers ; Humans ; Hydrogen-Ion Concentration ; Magnetic Resonance Spectroscopy* ; Phosphocreatine ; Phosphorus ; Spectrum Analysis

Attapulgite(ATP), as a fertilizer slow-release agent and soil conditioner, has shown remarkable effect in improving the utilization rate of fertilizer and the yield and quality of agricultural products and Chinese medicinal materials. This study aims to explore the effect of ATP on the growth and root quality of Angelica sinensis. To be specific, Mingui 1 was used, and through the pot(soil culture) experiment in the Dao-di producing area, the effects of conventional chemical fertilizer added with ATP on the morphology, photosynthesis, soil respiration, and content of ferulic acid and volatile oil in roots of Mingui 1 were detected. The underlying mechanism was discussed from the perspective of source-sink relationship. The results showed that ATP, via the fertilizer slow-release effect, could meet the needs of A. sinensis for nutrients at the root expansion stage, improve the net photosynthetic rate of leaves and aboveground biomass of plants, and promote the transfer and accumulation of nutrients from the aboveground part(source) to the underground root(sink) in advance during the dry matter accumulation period of roots, so as to improve the root weight per plant. ATP can increase the content of total ferulic acid(the sum of free ferulic acid and coniferyl ferulate), the main effective component of Angelicae Sinensis Radix, by promoting the synthesis of ferulic acid in the roots and the transformation to coniferyl ferulate. However, it had little effect on the content of volatile oil. ATP had certain influence on soil respiration, which needs to be further explored from root activity, rhizosphere microorganisms, and soil microorganisms. This study can lay a basis for soil remediation and improvement and ecological cultivation of A. sinensis.
Adenosine Triphosphate ; Angelica sinensis/chemistry* ; Coumaric Acids ; Fertilizers/analysis* ; Magnesium Compounds ; Oils, Volatile/chemistry* ; Plant Roots/chemistry* ; Silicon Compounds ; Soil

PURPOSE: We evaluated the usefulness of the in vitro adenosine triphosphate-based chemotherapy response assay (ATP-CRA) for prediction of clinical response to fluorouracil-based adjuvant chemotherapy in stage II colorectal cancer. MATERIALS AND METHODS: Tumor specimens of 86 patients with pathologically confirmed stage II colorectal adenocarcinoma were tested for chemosensitivity to fluorouracil. Chemosensitivity was determined by cell death rate (CDR) of drug-exposed cells, calculated by comparing the intracellular ATP level with that of untreated controls. RESULTS: Among the 86 enrolled patients who underwent radical surgery followed by fluorouracil-based adjuvant chemotherapy, recurrence was found in 11 patients (12.7%). The CDR ≥ 20% group was associated with better disease-free survival than the CDR < 20% group (89.4% vs. 70.1%, p=0.027). Multivariate analysis showed that CDR < 20% and T4 stage were poor prognostic factors for disease-free survival after fluorouracil-based adjuvant chemotherapy. CONCLUSION: In stage II colorectal cancer, the in vitro ATP-CRA may be useful in identifying patients likely to benefit from fluorouracil-based adjuvant chemotherapy.
Adenocarcinoma ; Adenosine Triphosphate ; Adenosine* ; Cell Death ; Chemotherapy, Adjuvant* ; Colorectal Neoplasms* ; Disease-Free Survival ; Drug Screening Assays, Antitumor ; Drug Therapy* ; Fluorouracil ; Humans ; In Vitro Techniques* ; Multivariate Analysis ; Recurrence

BACKGROUNDPhosphorous magnetic resonance spectroscopy ((31)P-MRS) has been successfully applied to study intracellular membrane compounds and high-energy phosphate metabolism. This study aimed to evaluate the capability of dynamic (31)P-MRS for assessing energy metabolism and mitochondrial function in skeletal muscle from type 2 diabetic patients.

METHODSDynamic (31)P-MRS was performed on 22 patients with type 2 diabetes and 26 healthy volunteers. Spectra were acquired from quadriceps muscle while subjects were in a state of rest, at exercise and during recovery. The peak areas of inorganic phosphate (Pi), phosphocreatine (PCr), and adenosine triphosphate (ATP) were measured. The concentration of adenosine diphosphate (ADP) and the intracellular pH value were calculated from the biochemistry reaction equilibrium. The time constant and recovery rates of Pi, PCr, and ADP were analyzed using exponential curve fitting.

RESULTSAs compared to healthy controls, type 2 diabetes patients had significantly lower skeletal muscle concentrations of Pi, PCr and β-ATP, and higher levels of ADP and Pi/PCr. During exercise, diabetics experienced a significant Pi peak increase and PCr peak decrease, and once the exercise was completed both Pi and PCr peaks returned to resting levels. Quantitatively, the mean recovery rates of Pi and PCr in diabetes patients were (10.74 ± 1.26) mmol/s and (4.74 ± 2.36) mmol/s, respectively, which was significantly higher than in controls.

CONCLUSIONSNon-invasive quantitative (31)P-MRS is able to detect energy metabolism inefficiency and mitochondrial function impairment in skeletal muscle of type 2 diabetics.

Adenosine Diphosphate ; analysis ; Adenosine Triphosphate ; analysis ; Adult ; Diabetes Mellitus, Type 2 ; metabolism ; Female ; Humans ; Magnetic Resonance Spectroscopy ; methods ; Male ; Middle Aged ; Mitochondria, Muscle ; metabolism ; Muscle, Skeletal ; metabolism ; Phosphates ; analysis ; Phosphocreatine ; analysis ; Phosphorus ; chemistry

The KFDA (Korea Food & Drug Administration) has performed a collaborative toxicogenomics project since 2003. Its aim is to construct a toxicology database of 12 compounds administered to mice at initial phase. We chose 6-MP (6-mercaptopurine) which has been used in the treatment of childhood leukemia. It was administered at low (0.224 mg/kg) and at high (2.24 mg/kg) dose (5 mice per group) intraperitonealy to the postnatal 6 weeks mice, then the serum and liver were collected at the indicated time (6, 24 and 72 h) after scarification. Serum biochemical markers for liver toxicity were measured and histopathologic studies also were carried out. The gene expression profiling was carried out by using Applied Biosystems 1700 Full Genome Expression Mouse. By self-organization maps (SOM), we identified groups with unique gene expression patterns, some of them are supposed to be related to 6-MP induced toxicity, including lipid metabolism abnormality, inflammatory response, oxidative stress, ATP depletion and cell death. The potential toxic effects appearing as gene expression changes are dependent of the time of 6-MP but independent of the dosage of it. This study would contribute to establishment of international database as well as national one about hepatotoxicity.
Adenosine Triphosphate ; Animals ; Cell Death ; Gene Expression Profiling* ; Gene Expression* ; Genome ; Leukemia ; Lipid Metabolism ; Liver* ; Mice ; Microarray Analysis ; Oxidative Stress ; Toxicogenetics ; Toxicology ; Biomarkers

This study was performed to assess the accuracy of 31P magnetic resonance spectroscopy(MRS) in the evaluation of myocardial ischemia in cats. Twelve cats underwent myocardial ischemia and reperfusion induced by 90 minutes ligation followed by 90 minutes recirculation of the left anterior descending coronary artery (LAD). MRS was performed using a 4.7T Biospec MRS/MRI system (Bruker, Switzerland). An inner diameter 1.5cm-sized doubly tuned surface coli was used for the collection of the MR signal. The coli was implanted to the epicardial surface at the expected area of infarction. 31P MRS was acquired before and during the periods of ischemia and reperfusion with 5-minute to 30-minute of intervals. After completion of the 31P MRS study, animals were sacrificed and the hearts were excised for 2,3,5-triphenyl tetrazolium chloride (TTG0 histochemical staining. The area of infarct was measured on the photographs of TTG stained heart slices using a computer programmed planimetry and the results were compared with those of the 31P MRS study. The level of phosphocreatine (PCr) was decreased to 28.2±6.9% of the baseline level 90 minutes after occlusion and recovered to 43.8±4.8% of the baseline level at the end of the reperfusion. A 50% depletion of PCr was reached 5 minutes after the LAD occlusion. The ATP was decreased to a 26.6±3.6% of the baseline level 90 minutes after occlusion and recovered to a 35.9±6.0 of the baseline level 90 minutes after reperfusion. The decreasing rate of ATP was slower than that of PCr showing a 50% of depletion 15 minutes after occlusion. The PCr/ATP ratio was 1.16±0.09 at the baseline, decreased to 0.88±0.07 at 30 minutes of occlusion, and then progressively increased during the late ischemic and reperfused periods. The ratio of the infarcted area to the effective signal area of the surface coli was inversely correlated to the ATP (r=0.68) and PCr (r=0.40) levels obtained at the end of reperfusion. In conclusion, 31P MRS reflects the changes in myocardial high energy phosphorous metabolism during the actue ischemia and reperfusion. If on adequate localization technique is feasible, 31P MRS can be used clinically in the diagnosis and monitoring of the patients with acute myocardial infarction.
Adenosine Triphosphate ; Animals ; Cats* ; Coronary Vessels ; Diagnosis ; Heart ; Humans ; Infarction ; Ischemia ; Ligation ; Metabolism ; Myocardial Infarction* ; Myocardial Ischemia ; Phosphocreatine ; Polymerase Chain Reaction ; Reperfusion ; Spectrum Analysis*

OBJECTIVETo investigate (1) whether ischemic preconditioning (IPC) could protect immature rabbit hearts against ischemia-reperfusion injury and (2) the role of K(ATP) channel in the mechanism of myocardial protection. Since cardioplegia is a traditional and effective cardioprotective measure in clinic, our study is also designed to probe the compatibility between IPC and cardioplegia.

METHODSNew Zealand rabbits aged 14 - 21 days weighing 220 - 280 g were used. The animals were anesthetized and heparinized. The chest was opened and the heart was quickly removed for connection of the aorta via Langendorff's method within 30 s after excision. All hearts were perfused with Krebs-Henseleit buffer balanced with gas mixture (O(2):CO(2) = 95%:5%) at 60 cm H(2)O (perfusion pressure). IPC consisted of 5 min global ischemia plus 10 min reperfusion. Glibenclamide was used as the K(ATP) channel blocker at a concentration of 10 micro mol/L before IPC. Cardiac arrest was induced with 4 degrees C St. Thomas cardioplegic solution, at which point the heart was made globally ischemic by withholding perfusion for 45 min followed by 40 min reperfusion. Thirty immature rabbit hearts were randomly divided into four groups: CON (n = 9) was subjected to ischemia-reperfusion only; IPC (n = 9) underwent IPC and ischemia-reperfusion; Gli (n = 6) was given glibenclamide and ischemia-reperfusion; and Gli + IPC (n = 6) underwent glibenclamide, IPC and ischemia-reperfusion. Coronary flow (CF), HR, left ventricle developed pressure (LVDP), and +/- dp/dt(max) were monitored at equilibration (baseline value) and 5, 10, 20, 30 and 40 min after reperfusion. The values resulting from reperfusion were expressed as a percentage of their baseline values. Arrhythmia quantification, myocardial enzyme in the coronary effluent and myocardial energy metabolism were also determined.

RESULTSThe recovery of CF, HR, LVDP and +/- dp/dt(max) in preconditioned hearts was best among the four groups. The incidence of arrhythmia was low and less CK-MB leaked out in the IPC group. Myocardial ATP content was better preserved by IPC. Pretreatment with glibenclamide completely abolished the myocardial protection provided by IPC, but did not affect ischemia-reperfusion injury.

CONCLUSIONSWhile applying cardioplegia, IPC provides significant cardioprotective effects. Activation of K(ATP) channels is involved in the mechanism of IPC-produced cardioprotection.

Adenosine Triphosphate ; analysis ; Animals ; Creatine Kinase ; secretion ; Creatine Kinase, MB Form ; Heart Arrest, Induced ; Hemodynamics ; Ischemic Preconditioning, Myocardial ; Isoenzymes ; secretion ; Potassium Channels ; physiology ; Rabbits