The aim of this study was to optimize methods for quantifying 13 uridine 59-diphosphate-glucuronosyltransferase (UGT) isoforms (UGT1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B10, 2B15, and 2B17) in human liver, intestinal, and kidney microsomes, and in recombinant human UGT-expressing insect cell membranes (rhUGTs) by targeted peptide-based quantification using liquid chromatography-tandem mass spectrometry. Production of targeted peptides was compared by combining three denaturing agents (urea, sodium deoxycholate, and octyl glucoside) and three denaturing temperatures (37°C, 60°C, and 95°C) followed by tryptic digestion for 2-20 hours. Denaturing conditions and digestion times yielding high production efficiency varied markedly among isoforms and specimens, indicating the importance of specific optimization. Each UGT isoform was quantified using the methods found to be optimal.
BackgroundAll clinical and biological manifestations related to postcardiac arrest (CA) syndrome are attributed to ischemia–reperfusion injury in various organs including brain and heart. Molecular hydrogen (H2) has potential as a novel antioxidant. This study tested the hypothesis that inhalation of H2 gas starting at the beginning of cardiopulmonary resuscitation (CPR) could improve the outcome of CA.Methods and ResultsVentricular fibrillation was induced by transcutaneous electrical epicardial stimulation in rats. After 5 minutes of the subsequent CA, rats were randomly assigned to 1 of 4 experimental groups at the beginning of CPR: mechanical ventilation (MV) with 2% N2 and 98% O2 under normothermia (37°C), the control group; MV with 2% H2 and 98% O2 under normothermia; MV with 2% N2 and 98% O2 under therapeutic hypothermia (TH), 33°C; and MV with 2% H2 and 98% O2 under TH. Mixed gas inhalation and TH continued until 2 hours after the return of spontaneous circulation (ROSC). H2 gas inhalation yielded better improvement in survival and neurological deficit score (NDS) after ROSC to an extent comparable to TH. H2 gas inhalation, but not TH, prevented a rise in left ventricular end-diastolic pressure and increase in serum IL-6 level after ROSC. The salutary impact of H2 gas was at least partially attributed to the radical-scavenging effects of H2 gas, because both 8-OHdG- and 4-HNE-positive cardiomyocytes were markedly suppressed by H2 gas inhalation after ROSC.ConclusionsInhalation of H2 gas is a favorable strategy to mitigate mortality and functional outcome of post-CA syndrome in a rat model, either alone or in combination with TH.
t is widely recognized that accumulation of abdominal visceral fat is strongly related to the development of coronary artery disease (CAD). [1][2][3][4][5][6] Epicardial adipose tissue (EAT) is the actual visceral fat of the heart deposited under the visceral layer of the pericardium and has the same origin as abdominal visceral fat. Pathological investigations revealed EAT and the adventitia of coronary arteries or myocardium to be contiguous, with no intervening structures. 7 The accumulation of EAT shows a good correlation with the volume of abdominal visceral fat and EAT is also known to be a rich source of free fatty acids and a number of bioactive molecules and inflammatory cytokines. [8][9][10][11] Some reports have suggested a crucial role of EAT in the development of CAD through changes in adipokine expressions in EAT, which promote pro-inflammatory characteristics, thereby possibly facilitating the progression of coronary atherosclerosis. [9][10][11] In fact, de Vos et al have reported that peri-coronary EAT thickness is strongly related to vascular risk factors and coronary calcification in post-menopausal women. 12 A recent study demonstrated that 64-slice multidetectorrow computed tomography (MDCT) is suitable for volumetric quantification of EAT with higher reproducibility than measurements of EAT thickness by echocardiography, and that excessive accumulation of EAT was associated with obesity and metabolic syndrome. 13 MDCT provides noteworthy information about coronary arteries including not only the presence and degree of stenotic lesions but also of subclinical atherosclerotic plaques. 14-19 MDCT can identify atherosclerotic plaques, in vessels with only minimal angiographic disease, with high sensitivity and moderate specificity as compared with intravascular ultrasound (IVUS). Moreover, MDCT can detect significant atherosclerotic plaques in vessels with signs of positive remodeling, which tend to be underestimated by conventional coronary angiography (CAG). 20,21 We sought to determine the relationship between the epicardial fat volume and the severity and extent of atherosclerosis of the whole coronary tree using CAG and MDCT in patients presenting with a possible diagnosis of stable effort angina. The relationship between the epicardial fat volume measured by 64-slice multidetector computed tomography (MDCT) and the extension and severity of coronary atherosclerosis was investigated. Methods and Results: Both MDCT and conventional coronary angiography (CAG) were performed in 71 consecutive patients who presented with effort angina. The volume of epicardial adipose tissue (EAT) was measured by MDCT. The severity of coronary atherosclerosis was assessed by evaluating the extension of coronary plaques in 790 segments using MDCT data, and the percentage diameter stenosis in 995 segments using CAG data. The estimated volume of EAT indexed by body surface area was defined as VEAT. Increased VEAT was associated with advanced age, male sex, degree of metabolic alterations, a history of acute corona...
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