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Animal, Human, and in Vitro Test Methods for Predicting Skin Irritation
Published in Francis N. Marzulli, Howard I. Maibach, Dermatotoxicology Methods: The Laboratory Worker’s Vade Mecum, 2019
Sunita M. Patil, Esther Patrick, Howard I. Maibach
In order to distinguish between mild and moderate irritants in an acute exposure test, Finkelstein and his colleagues (1963, 1965) used pretreatment of test sites with an irritant and enhanced visualization of the response by injection of trypan blue in order to increase test sensitivity. The technique was performed in anesthetized rabbits, rats, or guinea pigs. A circular area of the shaved abdomen was painted with a 20% solution of formaldehyde that was allowed to dry for 5 min. This was repeated three times and then 1 -in cotton flannel pads were saturated with test material and applied to each site. A control substance of known irritancy was tested in each study. Pads were secured in place and the entire trunk was wrapped in polyethylene. A solution of trypan blue was injected into subcutaneous tissue away from the dosage sites. The dye was absorbed and served as a marker for plasma leakage because it spontaneously binds to albumin. After 16 h, patches were removed and the degree of bluing at each site was evaluated on a 0–100% scale. In light of more recent work comparing the reactivity of dorsal and abdominal animal skin (Vinegar, 1979), one wonders if the enhanced sensitivity was due in part to choice of test site.
Metals
Published in Frank A. Barile, Barile’s Clinical Toxicology, 2019
Anirudh J. Chintalapati, Frank A. Barile
Industrially, Fe is used in powder metallurgy and as a catalyst in chemical reactions. Steel is one of the most important alloys of iron and is incorporated into construction materials. Ferric ferrocyanide, a dark blue, amorphous solid formed by the reaction of potassium ferrocyanide with a ferric salt (Prussian blue), is used as a pigment in paint and in laundry bluing. Potassium ferricyanide (red prussiate of potash) is obtained from ferrous ferricyanide (Turnbull’s blue) and is integrated into blueprint paper.
Circulation of fluid between plasma, interstitium and lymph
Published in Neil Herring, David J. Paterson, Levick's Introduction to Cardiovascular Physiology, 2018
Neil Herring, David J. Paterson
Most chemical mediators of inflammation act primarily on receptors expressed by the postcapillary venule to cause endothelial gap formation (Figures 11.26 and 11.27). The endothelial gaps create holes in the normally continuous glycocalyx, disrupting the semipermeable barrier. Consequently, water and plasma proteins leak out rapidly into the tissue (Figure 11.28). The greater the inflammatory stimulus, the greater the number of the leakage spots. In ischaemia-reperfusion injury (Section 6.12), glycocalyx damage may possibly raise permeability without gap formation. A common pharmacological test for inflammation, the skin blueing test, is based on the raised protein permeability. The dye Evans blue is injected into the plasma of a rat. Little escapes into normal skin because the dye binds to circulating plasma albumin; however, if an inflammatory agent is injected into the skin, the dye-albumin complex leaks out, producing a blue skin patch.
Acutely increased aquaporin-4 exhibits more potent protective effects in the cortex against single and repeated isoflurane-induced neurotoxicity in the developing rat brain
Published in Toxicology Mechanisms and Methods, 2023
Habip Yılmaz, Aslıhan Şengelen, Serdar Demirgan, Hüsniye Esra Paşaoğlu, Melike Çağatay, İbrahim Emre Erman, Mehmet Bay, Hasan Cem Güneyli, Evren Önay-Uçar
Immunohistochemical reactions were performed using an automated immunohistochemical stainer (Ventana BenchMark ULTRA, Ventana Medical Systems, Inc., Tucson, AZ, USA). The 2-µm-tissue sections (of hippocampus, cerebellum, and cortex) were deparaffinized using the EZ prep solution (Ventana), incubated with antigen retrieval buffer (CC1 solution, Ventana) at 98 °C for 60 min. Endogenous peroxidase activity was blocked with 3% hydrogen peroxide for 4 min at 37 °C (UltraView Universal DAB Inhibitor, Ventana). Slides were incubated with the following primary antibodies (at 37 °C for 60 min): anti-AQP4 (1:250, sc-20812, Santa Cruz Biotechnology, Dallas, TX, USA), and anti-4HNE (1:50, MAB3249, R&D Systems, Minneapolis, MN, USA). Detection was performed using the Ventana ultraView DAB Detection kit (Ventana Medical Systems, Inc.) which uses a hydrogen peroxide substrate and diaminobenzidine tetrahydrochloride chromogen. Final counterstain was performed with Ventana hematoxylin and bluing reagent. Slides were washed with Tris buffer (pH 7.6), mounted using a xylene-based mounting medium, and visualized using light microscopy (Olympus BX51, Tokyo, Japan). IHC analysis was made by a single pathologist blinded to the study protocol. IHC score for 4HNE was calculated using percent positive cells. Quantification for AQP4 levels was performed using Fiji software (ImageJ v2) as described by Patera et al. (2019). Data were presented as the mean optical density (MOD; = log (max intensity/mean intensity)].
Feasibility of olfactomedin 4 as a molecular biomarker for early diagnosis of gastric neoplasia after intestinal metaplasia
Published in Scandinavian Journal of Gastroenterology, 2023
Lixing Pang, Xin Yan, Dongxing Su, Xianbin Wu, Haixing Jiang
Paraffin-embedded tissue sections were successively baked, dewaxed in graded xylene, hydrated with anhydrous alcohol, dehydrated in gradient alcohol, and antigen-repaired in a sodium citrate solution. A phosphate-buffered saline (PBS) washing process was conducted three times, each for 3 min. Then, the tissue sections were blocked using H2O2 for 10 min and washed three times in a PBS solution (3 min each time). Anti-OLFM4 primary antibodies (1:100) were added at room temperature for 3 h (16–25 °C), followed by a secondary antibody (chromogen) at room temperature for another 20 min. Triple PBS washing was performed before and after hybridization. The color was developed with DAB for 8 min. This was followed by washing with water, hematoxylin staining, rapid differentiation in acid alcohol, bluing in lithium carbonate, exposure to anhydrous alcohol, drying, and sealing with resin.
SERPINB9 is commonly amplified and high expression in cancer cells correlates with poor immune checkpoint blockade response
Published in OncoImmunology, 2022
Sofía Ibáñez-Molero, Alex van Vliet, Joanna Pozniak, Karlijn Hummelink, Alexandra M. Terry, Kim Monkhorst, Joyce Sanders, Ingrid Hofland, Ewout Landeloos, Yannick Van Herck, Oliver Bechter, Thomas Kuilman, Weiwei Zhong, Jean-Christophe Marine, Lodewyk Wessels, Daniel S. Peeper
Immunohistochemistry of samples was performed on a BenchMark Ultra autostainer (Ventana Medical Systems). Briefly, paraffin sections were cut at 3 µm, heated at 75°C for 28 minutes and deparaffinized in the instrument with EZ prep solution (Ventana Medical Systems). Heat-induced antigen retrieval was carried out using Cell Conditioning 1 (CC1, Ventana Medical Systems) for 64 minutes at 95°C. PI-9 (SERPINB9) was detected using a polyclonal (1/1600 dilution, 1 hour at R.T, Abcam ab36624). Bound antibody was visualized using the OptiView DAB Detection Kit (Ventana Medical Systems). Slides were counterstained with Hematoxylin and Bluing Reagent (Ventana Medical Systems). The scoring was performed blinded with Slidescore®. Percentage of positivity (of any intensity) of the cancer cells or of the stroma (including lymphocytes, macrophages, fibroblasts, etc) was evaluated. Necrotic areas, luminal debris or other pigment areas (e.g., iron pigment or anthracosis) were not included. Samples with no tumor cells were excluded from the analysis.