Computational protocol: Pulmonary Edema Due to Oral Gavage in a Toxicological Study Related to Aquaporin-1, -4 and -5 Expression

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Protocol publication

[…] At the end of the test, which lasted 14 days for the acute test and 90 days for the sub-chronic test, surviving rats were euthanized by overdose inhalation of carbon dioxide. All tested rats were subjected to gross necropsy. The lungs and hearts were removed and fixed in 10% neutral buffer formalin for 48 h. Fixed specimens were embedded in paraffin, cut into 4-µm sectioned and then therefore stained with hematoxylin and eosin (H&E).The lungs were examined histopathological as follows: alveolar septum thickness was measured as the ratio of the septal area to the alveolar sac, while peribronchial and perivascular cuffing, perivascular edema, alveologenic edema, tunica media hyperplasia of the pulmonary artery, mast cell accumulation and accumulation of alveolar lipid containing macrophage were evaluated by severity scoring. The severity scores were classified as follows: absent, 0; focal, 1; moderate, 2; and severe, 3. Moreover the hearts were examined histopathologically for the early stage of cardiomyopathy based on aggregation of mononuclear inflammatory cells with fibrosis, which was also graded by method above. Area measurements were performed using an image analysis program (ImageJ®, NIH, version 1.36, by Wayne Rassband) with 10 fields of the area of interested/sample. [...] Quantitative results were expressed as the mean ± standard error of mean. Data were statistically analyzed with IBM® SPSS® Statistics software version 20 using a one-way analysis of variance (ANOVA) followed by Levene’s test. To differentiate differences between groups, multiple comparisons with the Bonferroni test and Dunnett’s test were performed for equal and non-equal variance assumptions, respectively. Bivariate correlation was examined by Spearman’s rho correlation test. Proportional statistics were examined by Chi-square test. […]

Pipeline specifications

Software tools ImageJ, SPSS
Applications Miscellaneous, Microscopic phenotype analysis
Diseases Pulmonary Edema, Lung Injury, Drug-Related Side Effects and Adverse Reactions