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Background--Endothelial dysfunction may contribute to the risk of premature atherosclerosis in patients with diabetes. Endothelin ET-1 ; may be involved in this process by activating smooth muscle cell mitogenesis and leukocyte adhesion. We sought to assess the activity of endogenous ET-1 in a group of patients with type II diabetes mellitus with the use of antagonists of ET-1 receptors. Methods and Results--Forearm blood flow FBF ; responses strain gauge plethysmography ; to intraarterial infusion of a selective blocker of ETA receptors BQ-123 ; and, on a different occasion, to ET-1, were measured in 15 patients with diabetes and 12 healthy controls. In addition, 5 patients with diabetes received coinfusion of BQ-123 and BQ-788 a selective blocker of ETB receptors ; . In normal subjects, BQ-123 did not significantly modify FBF from baseline P 0.16 ; . In contrast, BQ-123 administration resulted in a significant vasodilator response in patients with diabetes P 0.001 ; . Infusion of exogenous ET-1 resulted in lower vasoconstrictor responses in patients with diabetes than in controls P 0.001 ; , whereas the vasoconstrictor response to norepinephrine was similar in the 2 groups P 0.78 ; . In patients with diabetes, the vasodilator response to selective ETA blockade was not significantly modified by nonselective blockade of ET-1 receptors obtained by coinfusion of BQ-123 and BQ-788. Conclusions--The activity of endogenous ET-1 on ETA receptors is enhanced in the resistance vessels of patients with diabetes, whereas their sensitivity to exogenous ET-1 is blunted. This abnormality may participate in the pathophysiology of vascular complications associated with diabetes. Circulation. 2002; 106: 1783-1787. ; Key Words: endothelin diabetes vasculature receptors atherosclerosis.
Energy Intensity has improved 22 percent since 1994 baseline, exceeding the the 20 percent improvement goal. In 2006 Dow defined a new 10 year energy goal to drive a 25% improvement in energy intensity from the 2004 baseline. The measurement system for tracking from 2004 to 2015 was improved making it inappropriate to directly compare historical EI numbers with the information below.
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Products 20X ; and Taqman Master Mix 2X ; from Applied Biosystems were used to amplify perforin and RNAse P mRNAs according to the instruction manual. Reactions were run on ABI Prism 7700 Sequence Detection System Applied Biosystems ; in a final volume of 25 L for 50 cycles. Perforin expression was normalized to RNAse P levels in each sample. Cytolytic activity PBMCs exposed or not to rIL-2 ; and polyclonal rIL-2activated NK cells were tested for cytolytic activity against various NK-susceptible tumor target cells including K562 and LCL 721.221 in a 4-hour 51Cr-release assay as previously described.30 In other experiments their cytolytic activity was evaluated against autologous or heterologous PHA blasts obtained as previously described.31 The concentration of anti-HLA class I mAb used for these experiments was 10 g mL. The effector-target E T ; ratios are indicated in the figure legends. Immunofluorescence staining PMN and NK cells were obtained as reported in "PBMCs, NK cells, and neutrophil purification and culture." Cells were counted and used for cytospin preparations. Slides were air-dried and used for immunofluorescence staining. Primary antibodies included in this study included anti human NE mAb clone NP57 at dilution 1: 25 Dako Cytomation, Glostrup, Denmark ; , anti-NE rabbit polyclonal Calbiochem ; at dilution 1: 300, antihuman perforin 1: 500, clone dG9; Becton Dickinson, Oxford, United Kingdom ; , rabbit anticathepsin D Upstate Biotechnology, Lake Placid, NY ; , and rabbit antiLamp-2 AS120, 1: 200 ; and a pool of antihuman granzyme B clone GRB7, 1: 20, Valter Occhiena, Milan, Italy; clone GZB01, 1: 20, Biocarta, Hamburg, Germany; clone 11F1, 1: 20, Novocastra, Newcastle upon Tyne, United Kingdom ; . For double immunofluorescence of PMN cells, rabbit antihuman elastase and mouse anti-CD43 1: 25, clone MT1; Novocastra ; were used; elastase was revealed with swine antirabbit FITC-conjugated antibody 1: 25; Dako Cytomation ; and CD43 with Texas-Redconjugated goat antimouse IgG1 1: 75; Southern Biotechnology Associates ; . For immunofluorescence of NK cells confocal microscopy was performed as described.1 Briefly, cells were attached to glass slides in serum-free medium for 10 to 20 minutes, fixed in 20C methanol for 5 minutes, rinsed in PBS, preblocked in PBS, 1% normal goat serum for 15 minutes, and incubated with primary antibodies in PBS, 1% BSA for 40 minutes at room temperature. Cells were washed 3 to 4 times in PBS, 1% BSA and incubated with secondary antibody for 30 minutes at room temperature, washed 5 to 6 times in PBS, 1% BSA, and mounted in PBS, 90% glycerol 2.5% DABCO.
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