ABSTRACT
The Phospholipase D (PLD) superfamily is linked to neurological disease, cancer, and fertility, and a recent report correlated a potential loss-of-function PLD2 polymorphism with hypotension. Surprisingly, PLD2 -/- mice exhibit elevated blood pressure accompanied by associated changes in cardiac performance and molecular markers, but do not have findings consistent with the metabolic syndrome. Instead, expression of endothelial nitric oxide synthase (eNOS), which generates the potent vasodilator nitric oxide (NO), is decreased. An eNOS inhibitor phenocopied PLD2 loss and had no further effect on PLD2 -/- mice, confirming the functional relationship. Using a human endothelial cell line, PLD2 loss of function was shown to lower intracellular free cholesterol, causing upregulation of HMG Co-A reductase, the rate-limiting enzyme in cholesterol synthesis. HMG Co-A reductase negatively regulates eNOS, and the PLD2-deficiency phenotype of decreased eNOS expression and activity could be rescued by cholesterol supplementation and HMG Co-A reductase inhibition. Together, these findings identify a novel pathway through which the lipid signaling enzyme PLD2 regulates blood pressure, creating implications for on-going therapeutic development of PLD small molecule inhibitors. Finally, we show that the human PLD2 polymorphism does not trigger eNOS loss, but rather creates another effect, suggesting altered functioning for the allele.
Subject(s)
Blood Pressure/genetics , Nitric Oxide Synthase Type III/metabolism , Phospholipase D/deficiency , Signal Transduction , Animals , Cholesterol/metabolism , Gene Expression , Gene Expression Regulation , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Hydroxymethylglutaryl CoA Reductases/genetics , Hyperlipidemias/etiology , Hyperlipidemias/metabolism , Male , Mice , Mice, Knockout , Mutation , Nitric Oxide/biosynthesis , Nitric Oxide Synthase Type III/genetics , Obesity/etiology , Obesity/metabolismABSTRACT
This study aimed to conduct measurement uncertainty assessment of a new method for determination of Sudan colorants (Sudan I, II, III and IV) in food by high performance liquid chromatography (HPLC). Samples were extracted with organic solvents (hexane, 20% acetone) and first purified by magnesium trisilicate (2MgO·3SiO2). The Sudan colorants (Sudan I-IV) were also initially separated on C8 by gradient elution using acetonitrile and 0.1% (v/v) formic acid aqueous solution as the mobile phases and detected with diode-array detector (DAD). The uncertainty of mathematical model of Sudan I, II, III and IV is based on EURACHEM guidelines. The sources and components of uncertainty were calculated. The experiment gave a good linear relationship over the concentration from 0.4 to 4.0 μg/mL and spiked recoveries were from 74.0% to 97.5%. The limits of determination (LOD) were 48, 61, 36, 58 μg/kg for the four analytes, respectively. The total uncertainty of Sudan colorants (Sudan I, II, III and IV) was 810±30.8, 790±28.4, 750±27.0, 730±50.0 μg/kg, respectively. The recovery uncertainty was the most significant factor contributing to the total uncertainty. The developed method is simple, rapid, and highly sensitive. It can be used for the determination of trace Sudan dyes in food samples. The sources of uncertainty have been identified and uncertainty components have been simplified and considered.
Subject(s)
Humans , Azo Compounds , Chemistry , Chromatography, High Pressure Liquid , Methods , Food Analysis , Methods , Food Coloring Agents , Chemistry , Limit of Detection , Magnesium Silicates , Chemistry , Naphthols , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the value of multislice spiral computed tomography (MSCT) in demonstrating the relationship between bronchus and peripheral lung cancer.</p><p><b>METHODS</b>We prospectively performed volumetric targeted scans of 0.5 mm collimation with MSCT and reconstructed images of multiplanar reconstruction (MPR), curved multiplanar reformations (CMPR) and surface shaded display (SSD) in 53 peripheral lung cancers. The results were compared with macroscopic and microscopic specimens.</p><p><b>RESULTS</b>(1) The third- to seventh-order branches of the bronchi were clearly shown in all patients by the designed protocol. CT demonstrated the tumor-bronchus relationship in 29 (96.7%) adenocarcinomas and 13 (76.5%) squamous-cell carcinomas. Statistic analysis showed that there was no significant difference between the two groups (chi(2) = 2.8, P > 0.05). (2) The tumor-bronchus relationship was identified as four types with MSCT. Type I: bronchus was obstructed abruptly by the tumor, type II: bronchus penetrated into the tumor with tapered narrowing and interruption, type III: bronchus lumen shown within tumor was patent and intact, type IV: bronchus ran at the periphery of the tumor with intact or narrowed lumen. (3) Type I was shown in 31 of 53 (58.5%) tumors with squamous-cell carcinoma slightly more common than adenocarcinoma. Type II and type III were seen equally in 8 of 53 (15.1%) tumors which occurred only in adenocarcinomas. Type IV was seen in 15 of 53 (28.3%) tumors with adenocarcinoma being slightly more frequent than squamous cell carcinoma. (4) The tumor at the fourth-order bronchus was more common in squamous cell carcinoma, whereas that at the fixth-order bronchus was more likely in adenocarcinoma.</p><p><b>CONCLUSION</b>Volumetric targeted scan of ultra-thin section with MSCT and followed by MPR, CMPR and SSD reconstruction can greatly improve the manifestation of the bronchioles and accurately demonstrate the patterns of tumor-bronchus relationship, thereby reflecting pathologic changes to some extent.</p>
