PubMed Health⌕ Search

Biomedical subjects

K Nithipatikom

Publications and source records attributed to K Nithipatikom.

24 records · Page 2Linked to original sources

Metabolism of several 14C-nonylphenol isomers by rainbow trout (Oncorhynchus mykiss): in vivo and in vitro microsomal metabolites.

1. Gas chromatographic/mass spectroscopic analysis of a mixture of 14C-nonylphenols produced by alkylation of 14C-RUL-phenol with n-1-nonene indicated that the radio-synthesis produced three major isomers, 2-(4-hydroxyphenyl)-nonane, 3-(4-hydroxyphenyl)-nonane and 4-(4-hydroxyphenyl)-nonane. 2. Bile from rainbow trout exposed to a mixture of these isomers of 14C-nonylphenol was found to contain the glucuronic acid conjugates of three radiolabelled metabolites, which were more polar than their parent compounds. 3. Incubation of trout hepatic microsomes with NADPH and the 14C-nonylphenol isomers resulted in the production of three radiolabelled metabolites whose mobility on silica thin layer chromatography were similar to the deglucuronidated metabolites recovered from trout bile. 4. Metabolism of the 14C-nonylphenol isomers by trout hepatic microsomes was inhibited by omission of NADPH from the incubations as well as by addition of a P450 inhibitor, piperonyl butoxide to the incubations. 5. Analysis of the metabolites extracted from the microsomal incubations by gas chromatography/mass spectroscopy indicated that the parent isomers had been hydroxylated in the C-8 position on the nonane chain to give 2-(4-hydroxyphenyl)-8-hydroxynonane, 3-(4-hydroxyphenyl)-8-hydroxynonane and 4-(4-hydroxyphenyl)-8-hydroxynonane.

Animals↗

Simultaneous determination of nitrate and nitrite in biological samples by multichannel flow injection analysis.

An automated method for the simultaneous determination of nitrite and nitrate in biological samples by using a multichannel flow injection analyzer has been developed. The method was based on the reaction of nitrite with Greiss reagent. The sample solution was injected and equally divided into two channels; channel one (1) represented total nitrite obtained by cadmium reduction of nitrate to nitrite while channel two (2) represented only nitrite. The absorbance of the color product was measured by photometric detectors with 540-nm filters. This method combines high reproducibility of sample introduction via flow injection and sensitivity of spectrophotometric detection. The detection limit is 25 nM for both nitrite and nitrate. The chemistry manifolds are constructed of Teflon tubing which, along with a low-pressure Flowfit connector system, provides for low maintenance, ease of use, and high sample throughput. We demonstrated that the system can be used for the determination of both nitrate and nitrite in a variety of biological samples as well as a comparison of the results from this system and the HPLC system.

Animals↗

Bimakalim, an ATP-sensitive potassium channel opener, mimics the effects of ischemic preconditioning to reduce infarct size, adenosine release, and neutrophil function in dogs.

BACKGROUND: The primary goal of the present study was to determine whether the infarct size-reducing effect of preconditioning is associated with an increase in adenosine release from the ischemic myocardium during a prolonged occlusion period or the subsequent reperfusion period and by a decrease in neutrophil infiltration. A second objective was to determine whether bimakalim, a KATP channel opener, mimics the effects of ischemic preconditioning. METHODS AND RESULTS: Barbital-anesthetized open-chest dogs were subjected to 60 minutes of left anterior descending coronary artery (LAD) occlusion followed by 3 hours of reperfusion. In the preconditioning group, 5 minutes of LAD occlusion followed by 10 minutes of reperfusion was elicited before the 60-minute occlusion period. In two other groups, bimakalim 1 microgram/kg bolus followed by a 0.05 micrograms.kg-1.min-1 infusion or an equivalent volume of saline was administered intravenously 15 minutes before occlusion and continued until the time of reperfusion. In a final group, bimakalim was administered 10 minutes before reperfusion and continued until the end of the experiment. To measure the release of adenosine from the ischemic region, coronary venous blood samples were collected at various times during ischemia and after reperfusion, and the concentration of adenosine was measured. Myocardial infarct size was determined by triphenyl tetrazolium chloride; transmural myocardial blood flow, by radioactive microspheres. Transmural myeloperoxidase (MPO) activity, an index of neutrophil infiltration in the area at risk, was also measured. Preconditioning produced a marked reduction in infarct size (9.8 +/- 3.0% versus 28.6 +/- 5.2% in the control group, mean +/- SEM); adenosine release at 5, 10, 15, and 30 minutes of the 3-hour reperfusion period; and transmural MPO activity in the risk area. Similarly, pretreatment with bimakalim resulted in reductions in infarct size, adenosine release, and transmural MPO activity to an extent almost identical to that of preconditioning. When bimakalim was administered 10 minutes before reperfusion, the drug also produced a significant reduction in infarct size and transmural MPO activity; however, no significant reduction in coronary venous adenosine concentrations was observed. There were no significant differences in collateral blood flow between groups. CONCLUSIONS: These results indicate that myocardial preconditioning in the canine heart produced by a short period of ischemia or a KATP channel opener is not mediated by an increase in adenosine release, as measured by coronary venous adenosine concentrations, during 60 minutes of occlusion or the initial 30 minutes of reperfusion. A significant reduction in transmural MPO activity in the ischemic area also appears to result from KATP channel activation and may play a role, at least in part, in the reduction in infarct size observed, particularly when a KATP channel opener is administered just before reperfusion.

Adenosine↗

Effects of nicorandil and glyceryl trinitrate on infarct size, adenosine release, and neutrophil infiltration in the dog.

OBJECTIVE: The major aim of this study was to determine if nicorandil, a potassium channel opener nitrate, produces a reduction in myocardial infarct size at a non-hypotensive dose in dogs and to determine if this effect is the result of an increase in adenosine release or reduction in neutrophil infiltration into the ischaemic area. Glyceryl trinitrate was used for purposes of comparison. METHODS: Barbitone anaesthetised dogs were subjected to 60 min of left anterior descending coronary artery occlusion followed by 3 h of reperfusion. Nicorandil (100 microgram.kg-1 bolus followed by a 10 microgram.kg-1.min-1 infusion; NC/pre group), glyceryl trinitrate (10 microgram.kg-1 bolus followed by a 1 microgram.kg-1.min-1 infusion; GTN/pre group), or an equivalent volume of saline (control group) were given intravenously 15 min before occlusion and continued to the time of reperfusion. In two other groups, nicorandil (NC/post group) or glyceryl trinitrate (GTN/post group) were given 10 min before reperfusion and continued until the end of the experiment. To measure the release of adenosine from the ischaemic region, coronary venous blood samples were collected before occlusion, during occlusion, and at various times following reperfusion. Myocardial infarct size was determined by triphenyltetrazolium chloride and transmural myocardial blood flow by radioactive microspheres. Transmural myeloperoxidase activity, an index of neutrophil infiltration, was measured in biopsies obtained from the area at risk. RESULTS: Pretreatment with nicorandil and glyceryl trinitrate caused a marked reduction in myocardial infarct size expressed as percent of the area at risk [NC/pre group, 7.8(SEM 1.6)%; GTN/pre group, 11.9(2.3)%; control group, 31.0(5.6)%]. When nicorandil and glyceryl trinitrate were given before reperfusion, both drugs still produced a significant reduction in infarct size [NC/post group, 13.8(2.0)%; GTN/post group, 18.9(4.3)%]. Coronary venous adenosine concentrations during reperfusion were significantly lower in both nicorandil and glyceryl trinitrate pretreated groups, but not in the post-treated groups. Transmural myeloperoxidase activity was significantly lower in both nicorandil treated groups. CONCLUSIONS: Pretreatment with a non-hypotensive dose of nicorandil or glyceryl trinitrate markedly reduces myocardial infarct size and adenosine release from the ischaemic-reperfused area. These agents were also effective, but to a lesser degree, when given just before reperfusion. The cardioprotective actions of nicorandil appear to be related not only to its potassium channel opening activity but also in part to its nitrate activity.

Adenosine↗

Determination of plasma adenosine by high-performance liquid chromatography with column switching and fluorometric detection.

A new method for the rapid determination of plasma adenosine concentrations was developed by using high-performance liquid chromatography with a column switching technique and fluorometric detection. Several "stop solutions" were used to prevent the enzymatic degradation and cellular uptake and release of adenosine in blood samples. Red blood cells and certain denatured proteins were separated by centrifugation. Subsequently, the supernatant was transferred directly into autosample vials and adenosine was reacted with chloroacetaldehyde to form a strong fluorescent, 1-N6-ethenoadenosine. The adenosine derivative was injected directly and separated on a shielded hydrophobic phase column coupled with a C18 reverse-phase column using a column switching valve. Macromolecules and other interfering substances were excluded by the shielded hydrophobic phase column and bypassed to waste. Then, the adenosine derivative and other retained compounds were switched onto the reverse-phase column for further separation and subsequently to the fluorescence detector. The system reduces the analysis time and contamination of the column and hence allows a shorter cleanup time and a longer column lifetime. Adenosine as low as 30 fmol (signal-to-noise ratio, S/N = 3) can be detected by this method. The percentage of recovery of adenosine in plasma treated with adenosine deaminase was above 90%. This method is very rapid (without tedious sample preparation) and sensitive for determining adenosine in canine blood and should prove to be useful in analyzing the effects of ischemia and reperfusion on arterial and coronary venous adenosine concentrations in blood or perfusate samples released from the ischemic or hypoxic myocardium.

Adenosine↗