ABSTRACT
The Papanicolaou stain is a gold-standard staining method for tumor diagnosis in human cytology. However, it has not been used routinely in veterinary cytology, because of its complicated multistep procedure and requirement for wet fixation. Currently, a rapid Papanicolaou stain using air-dried smears is utilized in human cytology, but usefulness of this rapid-air-dry Papanicolaou (RAD-Pap) stain in the veterinary field has not been fully evaluated. The purpose of this study was to evaluate the usefulness of the RAD-Pap stain by using quantitative analysis. Air-dried impression smears were collected from tumor specimens and stained with RAD-Pap and Giemsa. Twelve parameters representing the criteria of malignancy were quantitated, and characteristics of the RAD-Pap were evaluated statistically. The RAD-Pap stain could be applied to all the smears, and images of nucleoli and chromatin patterns were clear and detailed. In quantitative analysis with the RAD-Pap stain, but not with the Giemsa stain, dispersion of nucleolus size and dispersion of nucleolus/nucleus ratio in malignant tumors were significantly higher than those in benign tumors. These findings demonstrated that the RAD-Pap stain was useful for obtaining detailed nuclear information, and the ability to differentiate benignity and malignancy by nucleolus findings was a principal advantage of this stain. This RAD-Pap stain could be routinely used as a supportive staining method in veterinary diagnostic cytology.
Subject(s)
Azure Stains , Cat Diseases/diagnosis , Dog Diseases/diagnosis , Neoplasms/veterinary , Vaginal Smears/veterinary , Animals , Cat Diseases/pathology , Cats , Cell Nucleolus/pathology , Cell Nucleus/pathology , Dog Diseases/pathology , Dogs , Neoplasms/diagnosis , Neoplasms/pathology , Vaginal Smears/methodsABSTRACT
The objective of this study was to establish a simple and rapid immunocytochemical technique that can be used in veterinary diagnostic cytology. Air-dried impression smears were collected from canine tumors. Samples of epithelial tumors, mesenchymal tumors, and malignant peripheral nerve sheath tumors and melanomas were used for detection of cytokeratin, vimentin, and S-100 protein, respectively. The labeled streptavidin-biotin system was used in the present study. Optimal fixation was determined using standard immunocytochemical procedures, and acetone fixation was found to be the most effective. Optimal concentrations of primary and secondary antibodies were determined at a preset 5-min incubation. Omission of H2O2 treatment, shortening the time for blocking and labeled-streptavidin incubation, and simplifying washing did not decrease immunopositive intensities or enhance false-positive reactions. The described rapid protocol requires approximately 45 min without the use of any special equipment.
Subject(s)
Dogs/metabolism , Immunohistochemistry/methods , Keratins/metabolism , S100 Proteins/metabolism , Vimentin/metabolism , Animals , AntibodiesABSTRACT
Hepatocyte growth factor (HGF) is a multifunctional growth factor with mitogenic, anti-apoptotic and anti-fibrotic activities. In this study, we investigated the effect of administration of recombinant human HGF on pulmonary arterial hypertension. Pulmonary arterial hypertension was induced in rats by a single injection of monocrotaline (MCT) and recombinant human HGF (0.12 mg/day) was administered into the right ventricle cavity using osmotic pumps, which were implanted subcutaneously 21 days after MCT injection. Continuous intravenous delivery of recombinant human HGF for 14 days led to prolonged survival of animals suffering from severe MCT-induced pulmonary arterial hypertension. Although a bolus injection of recombinant human HGF did not affect pulmonary arterial pressure, a 14-day administration of recombinant human HGF attenuated the inflammatory cell infiltrate, matrix accumulation and vascular medial thickening. As a consequence, the pulmonary lumen was enlarged and the pulmonary arterial pressure was significantly reduced. Additionally, continuous administration of recombinant human HGF suppressed lung tissue expression of platelet-derived growth factor, which plays an important role in the development of pulmonary arterial hypertension. These results indicate that recombinant human HGF possibly has a great potential for improving symptoms and altering the clinical course of pulmonary arterial hypertension.