novobiocin and Inflammation

Topics: Animals; Biomarkers; Caffeine; Carbamazepine; Corbicula; Energy Metabolism; Germ Cells; Humans; Ibuprofen; Inflammation; Neurotoxins; Novobiocin; Tamoxifen; Water Pollutants, Chemical

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

Cell-free, fat-free mammary secretions were tested in vitro for ability to support growth of streptococci associated with mastitis. Secretions were obtained prior to drying off, during the dry period, at calving, and during lactation from four cow treatment groups. Treatment groups were dry cow therapy, dry cow therapy and mammary glands subjected to induced inflammation 7 d post-drying-off, no dry cow therapy and no induced inflammation, no dry cow therapy but mammary glands subjected to induced inflammation. Growth of Streptococcus uberis, Streptococcus faecalis, and Streptococcus agalactiae in secretions from nonlactating glands was unaffected by induced inflammation. Growth of Streptococcus bovis was significantly inhibited in secretion obtained 14 d after induced inflammation. Dry cow therapy had no effect on streptococcal growth in secretion obtained 7 d after therapy. Streptococcal growth was greatest in secretions from involuted glands, and there was little or no evidence for growth inhibitory factors in cell-free, fat-free secretions obtained during the dry period. Milk from lactating glands inhibited streptococcal growth, and the inhibitory factor was presumptively identified as lactoperoxidase. Apolactoferrin, immunoglobulin, or both had little effect on streptococcal growth.

Topics: Animals; Apoproteins; Bacteriological Techniques; Body Fluids; Cattle; Citrates; Citric Acid; Culture Media; Disease Susceptibility; Female; Immunoglobulins; Inflammation; Lactation; Lactoferrin; Mammary Glands, Animal; Mastitis, Bovine; Milk; Novobiocin; Pregnancy; Streptococcal Infections; Streptococcus

An experimental product incorporating 500,000 IU of procaine penicillin G and 600 mg of sodium novobiocin in 2% aluminum monostearate-peanut oil gel (10-ml dose) was infused after the final milk-out at end of lactation into all 4 mammary quarters of 56 cows that were infected in at least 1 quarter. The therapeutic and prophylactic efficacies were published in the companion report. Infusion of the product in all quarters of 5 lactating cows resulted in only slight irritation. Penicillin was eliminated by the 11th milking and novobiocin by the 5th. After infusion in the dry udder, the antibiotics were no longer detectable in serous secretion after 14 days and failed to appear in urine at the earliest (7-day) sampling after administration. Neither antibiotic was detectable in the 1st postpartum milking after nonlactating periods as short as 3 weeks.

Topics: Animals; Cattle; Drug Combinations; Female; Inflammation; Lactation; Mammary Glands, Animal; Mastitis, Bovine; Novobiocin; Penicillin G Procaine; Pregnancy

Topics: Anti-Bacterial Agents; Chloramphenicol; Cortisone; Dexamethasone; Erythromycin; Female; gamma-Globulins; Gynecology; Humans; Inflammation; Novobiocin; Oxytetracycline; Pregnancy; Puerperal Infection; Tetracycline; Toxicology