metrizamide and Inflammation

This protocol describes microsphere-based protease assays for use in flow cytometry and high-throughput screening. This platform measures a loss of fluorescence from the surface of a microsphere due to the cleavage of an attached fluorescent protease substrate by a suitable protease enzyme. The assay format can be adapted to any site or protein-specific protease of interest and results can be measured in both real time and as endpoint fluorescence assays on a flow cytometer. Endpoint assays are easily adapted to microplate format for flow cytometry high-throughput analysis and inhibitor screening.

Topics: Animals; Biotinylation; Flow Cytometry; Fluorescence Resonance Energy Transfer; Green Fluorescent Proteins; High-Throughput Screening Assays; Humans; Inflammation; Kinetics; Microspheres; Peptide Hydrolases; Peptides; Reproducibility of Results; Temperature

Vasography is a common diagnostic procedure utilized in evaluating the male genital tract for causes of azoospermia, for evaluation of persistent perineal pain and seminal vesiculitis, and recently used in the staging of prostate cancer. In order to evaluate the effects of injectable agents on the healthy vas deferens, 7 groups of 10 rats had the left vas deferens injected by vasopuncture with saline, 4 contrast agents, doxycycline and absolute alcohol. The animals were sacrificed 8 weeks later, and were examined for patency of the vas and inflammatory response. Results revealed that vasopuncture did not produce obstruction. Vasography with water-soluble contrast media does entail a low risk of inflammation in the rat, but produces no obstruction. Intra-vas injection of doxycycline, injected by some for treatment of chronic seminal vesiculitis and perineal pain, is not deleterious in the rat model.

Topics: Animals; Contrast Media; Diatrizoate; Diatrizoate Meglumine; Doxycycline; Drug Combinations; Ethanol; Inflammation; Iophendylate; Male; Metrizamide; Radiography; Rats; Rats, Inbred Strains; Vas Deferens