Muscarinic receptor subtypes in equine tracheal smooth muscle

Selective muscarinic receptor antagonists were used to identify muscarinic receptor subtypes in equine trachealis strips. The M₁ receptor antagonist pirenzepine (10^–7 mol/L to 3 × 10^–5 mol/L) and the M₃ receptor antagonist 4-diphenylacetoxy-N-methylpiperidine (4-DAMP, 10^–9 mol/L to 3 × 10^–7 mol/L³) dose dependently inhibited the contractile responses to electrical field stimulation (EFS) and exogenous acetylcholine (ACh). Schild plots yielded a pA₂ value for pirenzepine vs ACh of 6.75 ± 0.09, which is consistent with the affinity for M₂ or M₃ receptors, and a pA₂ value for 4-DAMP vs ACh of 8.47 ± 0.09, which is in agreement with the affinity for M₃ receptors. The M₂ receptor antagonist gallamine (10^–5 mol/L and 10^–4 mol/L) did not affect the response of trachealis to exogenous ACh and low-frequency EFS (0.1–2 Hz) but decreased the responses to high-frequency EFS (4–16 Hz). These results suggest that the muscarinic receptors mediating contractions induced by ACh in equine tracheal smooth muscle are of the M₃ subtype. The lack of an increase in the response to EFS following gallamine suggests that functional prejunctional inhibitory M₂ receptors are not present on the cholinergic nerves innervating equine tracheal smooth muscle.     

Detection of TNF alpha receptors on ovine leucocytes by flow cytofluorimetry.

Tumour necrosis factor receptor activity was detected on sheep leucocytes using flow cytofluorimetry and commercially available fluorescent human tumour necrosis factor. The density of receptors per cell and the proportion of cells expressing TNF receptor activity was found to be in the following rank order: monocytes greater than granulocytes greater than lymphocytes.     

THE NON-CARDIAC DISTRIBUTION OF 99mTC-METHOXY-ISOBUTYL-ISONITRILE (SESTAMIBI) IN NORMAL BEAGLES

Technetium-99m methoxy-isobutyl-isonitrile (sestamibi) is a radiophartnaceutical used for the evaluation of myocardial perfusion. Increased uptake of sestamibi has also been documented in tumors. The objective of this study was to document the extracardiac biodistribution of ^99mTc sestamibi in the normal dog. Nine normal beagles were given 0.35 mCi/kg ^99mTc sestamibi intravenously, and 60 second images were made of the entire body at 1 hour post injection. A defined distribution pattern was recognized, with good visualization of the heart, liver, gallbladder, small intestine, kidneys, urinary bladder, popliteal lymph nodes, parotid salivary glands and zygomatic salivary glands. Splenic uptake was not seen.
Target to background ratios were calculated for all the regions listed, using background regions-of-interest with the smallest coefficient of variance for the denominator. The mean, range and standard deviation of these ratios are given.     

Comparison of dot blot hybridization, polymerase chain reaction, and virus isolation for detection of bovine herpesvirus-1 (BHV-1) in artificially infected bovine semen.

Bovine semen samples spiked with bovine herpesvirus 1 (BHV-1) were used to compare dot blot hybridization, polymerase chain reaction (PCR), and virus isolation for detection of BHV-1 in bovine semen. The PCR amplification used primers targeting the BHV-1 thymidine kinase gene and a nucleic acid releasing cocktail (GeneReleaser); the PCR product was used as the DNA probe in dot blot hybridization; virus isolation was done in primary bovine fetal testis (BFT) cell cultures. Semen diluted 1:20 in tissue culture medium had the least cytotoxicity and inhibition of viral cytopathic effects in BFT cells, allowing detection of 1 TCID50/100 microL of BHV-1 suspension by virus isolation. The presence of foreign DNA such as bovine sperm DNA or salmon sperm DNA increased the sensitivity of dot blot hybridization in detecting BHV-1, allowing detection of 20,000 TCID50/100 microL of neat semen. The inhibition of PCR amplification of BHV-1 DNA in bovine semen was eliminated by diluting the samples 1:20 in tissue culture medium. The best PCR amplification was obtained when semen was diluted 1:20 and when a reaction buffer of pH 9.0, with 1.0 mM MgCl2 was used. Under these conditions, the PCR followed by ethidium bromide staining of agarose gels could detect 1 TCID20/100 microL of sample, whereas PCR followed by Southern blot hybridization could detect 0.01 TCID50/100 microL of sample.(ABSTRACT TRUNCATED AT 250 WORDS)