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Ocular Irritation Testing
Published in David W. Hobson, Dermal and Ocular Toxicology, 2020
George P. Daston, F. E. Freeberg
Conquet et al.34 and Laillier et al.50 examined a number of different endpoints associated with edema of ocular tissues, including corneal and conjunctival water content as an index of swelling, and conjunctival and aqueous humor content of Evans blue dye to detect increased capillary permeability in the conjunctiva and iris and ciliary body. (Evans blue dye injected intravenously associates with plasma proteins. Its presence in extravascular spaces indicates extravasation of plasma proteins.) There was a strong correlation between conjunctival water content and Draize conjunctival score, and between corneal water content and Draize corneal score, at least for the first day after exposure to irritating solvents. There was also a good correlation between dye content of the conjunctiva and Draize conjunctival score. On the other hand, there was little relationship between dye concentration in the aqueous and visible iridial irritation.
Antiinflammatory Actions of VIP in the Lungs and Airways
Published in Sami I. Said, Proinflammatory and Antiinflammatory Peptides, 2020
In complementary studies, 3 μM of capsaicin (or solvent only, in control studies) was infused intratracheally (in 0.5 mL of solvent) in anesthetized, mechanically ventilated rats and guinea pigs. Ten minutes earlier, Evans blue was injected i.v. (30 mg/kg). A third group of animals received an i.v. infusion of VIP (20 μg/kg min), beginning 5 min before capsaicin, and for the rest of the experiment. Airway pressure and arterial BP were monitored throughout. At the end of the experiment, the trachea was removed, blotted clean, and extracted in formamide for spectrophotometric measurement of Evans blue at 620 nM. The dye content increased in tracheas from capsaicin-treated animals (to 22.8 ± 2.8 1JLg/g), but was at control value in tracheas from animals that also received VIP (p < .05).
Functions of the Kidneys and Functional Anatomy
Published in Peter Kam, Ian Power, Michael J. Cousins, Philip J. Siddal, Principles of Physiology for the Anaesthetist, 2020
Peter Kam, Ian Power, Michael J. Cousins, Philip J. Siddal
Approximately two-thirds of body weight is water. A 70-kg male has a total body water content of 42L, divided between the intravascular (3L plasma), interstitial (11L) and intracellular (28L) fluid compartments (the extracellular fluid is the sum of intravascular and interstitial volumes). Total body water can be estimated using dilution techniques with markers that diffuse throughout the total body water compartment, such as isotopically labelled water, using deuterium (2H) or tritium (3H). Markers used to determine the extracellular fluid must cross capillaries but not cell membranes; these include inulin, mannitol, radiosodium, radiochloride and thiosulphate (the latter is the most widely used). The intravascular fluid volume can be determined with a marker that remains within vessels, such as radiolabelled albumin, or the dye Evans blue, which binds to plasma albumin. The interstitial volume cannot be measured directly and is calculated by subtraction of the plasma volume from the extracellular volume.
Astragalus injection ameliorates lipopolysaccharide-induced cognitive decline via relieving acute neuroinflammation and BBB damage and upregulating the BDNF-CREB pathway in mice
Published in Pharmaceutical Biology, 2022
Ke Liu, Guoran Wan, Ruhong Jiang, Li Zou, Dong Wan, Huifeng Zhu, Shan Feng
Evans blue perfusion analysis was performed as described previously (Feng et al. 2018). The mice were anaesthetized with 10% chloral hydrate (0.1 mL/10 g, ip) and intracardially perfused with prewarmed 0.9% NaCl briefly to wash out blood cells, followed by 0.5% Evans blue in cold 4% paraformaldehyde. After perfusion, the brains were taken out, post-fixed in 4% paraformaldehyde at 4 °C for 4 h, and then cryoprotected in 20% and 30% sucrose solutions in phosphate-buffered saline (PBS) at 4 °C for 3 days. The brain tissues were cut into 30 μm thick sections in a cryostat (Leica Microsystems, Wetzlar, Germany). The sections were mounted on gel-coated slides, dried at 37 °C for 1 h, and kept at −20 °C for use. The Evans blue–perfused cerebral microvessels in the sections, which exhibited fluorescence when Evans blue bound to proteins, were examined and images were captured using a Leica fluorescence microscope equipped with a CCD camera (Leica Microsystems).
2,4,6-Trihydroxy-3-geranyl acetophenone suppresses vascular leakage and leukocyte infiltration in lipopolysaccharide-induced endotoxemic mice
Published in Pharmaceutical Biology, 2021
Yee Han Chan, Nazmi Firdaus Musa, Yi Joong Chong, Siti Arfah Saat, Faizul Hafiz, Khozirah Shaari, Daud Ahmad Israf, Chau Ling Tham
The permeability assay was performed based on previously described protocols with some modifications (Bae et al. 2012). A total number of 36 mice were used in this assay, where the mice were randomly allocated into six experimental groups (n = 6) as described in ‘Experimental animals’ section. Mice were pre-treated with PBS, tHGA (2, 20 and 100 mg/kg) or dexamethasone (3 mg/kg) via i.p. route for 1 h, followed by the induction of LPS (15 mg/kg) via i.p. route for 6 h. Evans blue (1%) was then injected intravenously (i.v.) and allowed to circulate in the blood vessels for 30 min. Next, the mice were sacrificed, and 2 mL of ice-cold PBS was injected into the mice peritoneal region to obtain a peritoneal wash. Permeated Evans blue was measured spectrophotometrically using a microplate reader (UVM 340, ASYS Hitech GmbH, Eugendorf, Austria) at a wavelength of 620 nm. The plate correction factor was fixed at 740 nm. The data were expressed in µg per mouse based on a standard curve.
Dioscin alleviates lipopolysaccharide-induced acute lung injury through suppression of TLR4 signaling pathways
Published in Experimental Lung Research, 2020
Chuntao Wang, Qingnian Li, Tianyu Li
ALI is characterized by a massive inflammatory cascade within the lungs in addition to severely impaired gas exchange, resulting from alveolar–capillary barrier disruption and pulmonary edema.1,3 In this study, the Evans blue dye extravasation assay (a marker for pulmonary capillary permeability) was employed to quantify the extent of alveolar–capillary barrier disruption. Compared with other treatments recommended for ALI, our study not only provided statistical data of Evans blue dye but also images of the Evans blue dye extravasation assay; obviously, the images were more convincing. Pretreatment with dioscin significantly diminished the pulmonary capillary permeability (Figure 3). To evaluate the magnitude of pulmonary edema, the lung W/D ratio was calculated. The results illustrated that pretreatment with dioscin decreased the lung W/D ratio (Figure 4), which indicates that dioscin could prevent the progression of pulmonary edema.