Evans blue

Several compounds that inhibit vesicular glutamate transport have been identified These include the dyes Evans Blue and Rose Bengal. In addition, the stilbene derivative 4,4 -diisothiocyanatostilbene-2,2 -disulfonic acid (DEDS), a compound commonly used as a specific inhibitor of anion channels, inhibits vesicular glutamate transport. Most known inhibitors have limited use as they are membrane impermeant, with the exception of Rose Bengal. 

Effect of the extracts on the increased vascular permeability induced by acetic acid in mice was determined according to Whittle method with some modifications [5]. Each test sample was administered orally to a group of ten mice, which were male Swiss albino mice (20-25 g) were purchased from the animal breeding laboratories of Refik Saydam Central Institute of Health (Ankara, Turkey), in 0.2 mF20 g body weight. Thirty minutes after the administration each mice was injected with 0.1 ml of 4% Evans blue (Sigma, St. Louis, Missouri, USA) in saline solution (/.v.) at the... 

Material Dose (mg/kg) Evans blue concentration (pg/ml) SEM Inhibition (%)... 

F. Blue 2B 250% (C.I. Direct Blue 6, C.I. No. 22610), Direct Brown BRL 200% (C.I. Direct Brown 95, C.I. No. 30145) Fabricolor Inc. Evans Blue (C.I. Direct Blue 53, C.I. No. 23860), Benzo Azurine G (C.I. Direct Blue 8, C.I. No. 24140) Pfaltz and Bauer Inc. Benzidine was obtained from Sigma Chemical Co., o-tolidine from Fisher Scientific Corp. and 3,3 -dimethoxybenzidine (o-dianisidine) from Eastman Kodak Company. Aniline, -aminophenol, -phenylenediamine and -nitroaniline used in the interference study were obtained from Chem Service Inc. 

Fig. 13.1 Cytoprotective action of adenosine in a quantitative model of mouse hindlimb ischemia and reperfusion (I/R) injury. Adult wild type mice were injected with (a) sterile vehicle (0.1% DMSO in phosphate-buffered saline, pH 7.4) or various adenosine receptor agonists (b, c) or antagonist (d, f). (a) Following ischemia and reperfusion, skeletal muscle showed a significant uptake of Evans Blue dye (EBD) in vehicle-treated mice (first part of (a), representative of 7 mice). The contra-lateral leg not subjected to ischemia-reperfusion showed virtually no EBD uptake. In the second part of (a), the same section was stained with rabbit polyclonal anti-skeletal muscle actin antibodies followed by staining with goat anti-rabbit IgG conjugated with FITC. (b) The nonselective adenosine agonist R-PIA caused a large reduction in the EBD-stained area (see both first and second parts to this figure, representative of six mice)...

Dhillon H. S., Donaldson D., Dempsey R. J., and Prasad M. R. (1994). Regional levels of free fatty acids and Evans blue extravasation after experimental brain injury. J. Neurotrauma 11 405 415. 

Figure 20.1 Distribution volume of Evans blue-labeled albumin in a rat fibrosarcoma as a function of (A) perfusion pressure or (B) perfusion rate. The ratio of distribution volume (Fd)/infused volume ( ] ) was quantified at the infusion pressures of 36, 50, 94, and 163 in cmH20, respectively. Symbols represent data from individual experiments N= 2 for pressure of 36 cmH20 and N=5 for other pressures. The line in (B) was obtained through linear curve-fitting of the data. Reproduced with permission (McGuire and Yuan, 2001).

Distribution volume of Evans blue-labeled albumin in a rat fibrosarcoma as a function of 400... 

Matsuda, R., Nishikawa, A., and Tanaka, H., 1995, Visualization of dystrophic muscle fibers in mdx mouse by vital staining with Evans blue evidence of apoptosis in dystrophin-deficient muscle, J Biochem (Tokyo), 118, pp 959-964. 

Experimentally, the BBB disruption can be assessed by the administration of dyes (e.g., Evans blue, Fluorescein) or radioactive tracers (e.g., [14C] sucrose) and visualization or measurement of their distribution in the brain parenchyma on tissue sections (Fig. 8.1). To estimate brain water content, the weight of a brain is determined with the wet-and-dry-weight method , i.e., weighing the tissue speci-... 

Fig.8.1a-d. Evans blue extravasation 6 h after permanent middle cerebral artery occlusion (MCAO) in Sprague-Dawley rats. Evans blue extravasation is shown in red and cell nuclei in blue labeled by 4 -6-diamidino-2-phenylindole (DAPI), a compound that binds to DNA. (a) A low-magnification view of the entire ischemic area, (b) Evans blue extravasation from a vessel located in the area adjacent to the ischemic area, (c) Electronic magnification of the box in (b) (arrow is pointing at a leakage of Evans blue from the vessel), (d) No extravasation of Evans blue is seen in a vessel from the same section seen in (b) and (c) but located in the corresponding region of the contralateral hemisphere Contra, contralateral. [Reproduced with permission from Yepes et al. (2003)]... 

Uyama O, Okamura N, Yanase M, Narita M, Kawabata K, Sugita M (1988) Quantitative evaluation of vascular permeability in the gerbil brain after transient ischemia using Evans blue fluorescence. J Cereb Blood Flow Metab 8 282-284... 

In fact, the D-protease was active it cleaved a particular synthetic substrate and was inhibited by specific inhibitors. To examine the structure of the D- and L-enzymes, Milton and coworkers tested both forms for activity with D and L forms of a chiral peptide substrate and for inhibition by d and l forms of a chiral peptide-analog inhibitor. Both forms were also tested for inhibition by the achiral inhibitor Evans blue. The findings are given in the table. 

Figure 8.6 shows a diagram of a general model of blood-tissue solute transport, used to analyze data on the transport of labeled solutes introduced in the blood or perfusate flow supplied to individual organs. The development and analysis of models of this sort to analyze solute transport in physiological systems is a field pioneered by Sangren and Sheppard [178], Renkin [172], and Crone [40], Optically detectable probes (such as Evans Blue dye bound to albumin) can be used in conjunction with model analysis to probe the intravascular transport of... 

---