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Biomedical subjects

C B Chapman

Publications and source records attributed to C B Chapman.

At least 19 recordsLinked to original sources

Nyctohemeral rhythm in the levels of S-adenosylmethionine in the rat pineal gland and its relationship to melatonin biosynthesis.

Liquid chromatographic techniques that permit the simultaneous analysis of S-adenosylmethionine, melatonin, and its intermediary metabolites N-acetyl-5-hydroxytryptamine and 5-hydroxytryptamine within individual pineal glands have been developed. S-Adenosylmethionine has been shown to undergo a marked nyctohemeral rhythm in the pineal gland of the rat, with maximal levels occurring during the light period and minimal levels during the dark period. Detailed studies of the temporal relationships between the levels of S-adenosylmethionine and those of melatonin and its intermediary metabolites suggest that an association exists between the levels of S-adenosylmethionine and the status of the biosynthesis of melatonin. Exposure of animals to continuous light and the administration of the beta-adrenoreceptor antagonist propranolol were both found to inhibit the induction of melatonin synthesis and prevent the reduction in the levels of S-adenosylmethionine during the dark period. As a corollary the induction of melatonin biosynthesis following the administration of the beta-adrenoreceptor agonist isoproterenol during the light period was accompanied by a marked decrease in the levels of S-adenosylmethionine in the pineal gland. The significance of the link between the nyctohemeral rhythms in the levels of S-adenosylmethionine and the biosynthesis of melatonin in the pineal gland is discussed in the context of the therapeutic efficacy of S-adenosylmethionine as an antidepressant.

Animals

The development of an enzyme-linked immunosorbent assay (ELISA) for cephalexin.

Cephalexin was structurally modified by the attachment of a spacer at the carboxylic acid through which it was subsequently covalently attached to BSA. This method permitted the molecule to be attached without cleavage of the beta-lactam ring giving a conjugate distinct from previously described immunogenic preparations of penicillins and cephalosporins. This approach required the development of a novel spacer molecule, and its synthesis and characterisation are reported. Rabbits were used to raise antisera and the antibodies produced were characterised with respect to their reactivity with cephalexin and various analogues, other cephalosporins, and a number of penicillins.

Animals

The role of procaine in adverse reactions to procaine penicillin in horses.

Procaine penicillin is a commonly used antibiotic in equine medicine but its use is associated with a substantial incidence of adverse reactions. Soluble procaine concentrations were determined by HPLC in several commercially available procaine penicillin preparations, including some that were involved in adverse reactions. The mean (+/- SEM) soluble procaine concentrations in the veterinary preparations was 20.18 +/- 5.07 mg/ml, which was higher than the concentration in the only procaine penicillin preparation for use in humans in Australia of 7.3 mg/ml. Heating the veterinary procaine penicillin preparations to 50 degrees C for 1 day led to a significant (P less than 0.01) increase in the amount of soluble procaine. Heating to 50 degrees C for 7 days also produced a significant (P less than 0.02) increase. Soluble procaine tended to return to baseline concentrations when veterinary procaine penicillin preparations were heated to 50 degrees C for 2 days then stored for 7 days at room temperature. Administration of procaine HCl intravenously (IV) at 2, 5, and 10 mg/kg produced behavioural, locomotor and vascular reactions, which were clinically similar to those reported in adverse reactions to procaine penicillin. The more severe reactions occurred at higher doses, although different horses responded variably at the same dose. Some adverse reactions lead to recumbency but none were fatal. The blood procaine concentrations 1 min after IV administration averaged 19.0 +/- 12.6 and 25.3 +/- 16 micrograms/ml at 2.5 mg/kg and 5 mg/kg, respectively. Ten min after administration, blood procaine concentrations were significantly higher (P less than 0.001) in the 5 mg/kg group than in the 2.5 mg/kg group. Intramuscular (IM) procaine HCl at 5 mg/kg produced significantly lower (P less than 0.001) blood concentrations than similar IV doses, and, in contrast to the IV doses, the amount of procaine in the blood was significantly higher 5 and 10 min after administration than it was after 1 min. Mild excitatory reactions in 4/5 horses were noted 5 to 10 min after IM administration. Administration of diazepam 20 s before procaine HCl prevented the excitatory adverse reaction in 2/2 horses, but administration after the procaine did not influence the outcome.

Animals

Detection of reserpine in horses by high-performance liquid chromatography.

A high-performance liquid chromatography (HPLC) assay was developed for the detection of reserpine. The assay was used to monitor the plasma concentrations of the drug given intramuscularly on one or two occasions to five horses. The blood concentrations of reserpine varied quite considerably between horses given the same dose of the drug. However, on average, reserpine could be detected consistently, and quantified, for 48 h after a single dose of 2.5 mg, and for a similar period after the second of two 2.5 mg doses given 13 d apart. Because of the apparently large variability in the pharmacokinetics of reserpine in horses, exact times cannot be given beyond which the drug will no longer be detectable in the plasma. However, following two doses of 2.5 mg reserpine given 13 d apart, at least 7 d must elapse after the second dose before there is no drug detectable in the plasma of most horses.

Animals

Adverse reaction to procaine penicillin G in horses.

Adverse reactions to intramuscular injections of procaine penicillin G are reported in 11 horses, five of which died. The clinical findings are presented and suggest central nervous involvement in most cases. Post mortem findings in one horse were consistent with anaphylaxis whereas in other cases the clinical findings, duration of treatment, speed of onset and subsequent completion of treatment supports diagnosis of an acute procaine toxicity syndrome.

Anaphylaxis

Development of an enzyme-linked immunosorbent assay for the detection of phenothiazine tranquillisers in horses.

An acepromazine (ACP) hapten was synthesised, coupled to bovine serum albumin and injected into a horse to produce antibodies to the drug. A competitive ELISA was developed whereby ACP attached to the solid phase via lysozyme competed with free ACP present in phosphate buffered saline, horse serum or horse urine for limiting amounts of antibody. The assay could detect the presence of ACP and, or, some of its metabolites in horse urine for at least 25 hours after intravenous injection of 0.1 mg kg-1 ACP maleate, but because of non-specific interference, horse serum could not be used. As little as 0.24 micrograms ml-1 ACP or its metabolites could be detected. The level of detection and the ease of performance of the assay make it an attractive alternative to the more complex methods currently available for the screening of horse urine samples at horse races, shows and sales.

Acepromazine

Proteolytic cleavage of immunoglobulin by enzymes released by Fasciola hepatica.

Immature Fasciola hepatica release a papain or cathepsin B-like proteolytic enzyme which cleaves immunoglobulins (Ig) of mouse, rat, rabbit and sheep in vitro. Mouse IgG and IgM molecules are both susceptible to cleavage as is hemoglobin. Whether single or multiple proteases are responsible for Ig cleavage is unknown. The proteolytic activity of secreted enzyme(s) is optimal at pH 3.5-4.5, but activity is also present at pH 7. Proteolysis is enhanced in the presence of 5 mM dithiothreitol or 100 mM cysteine. Based on studies with protease inhibitors, the F. hepatica enzyme activity has been identified as a thiol protease. It is destroyed by heating at 56 degrees C for 1 h, but retains activity after storage at -20 degrees C for 7 days. Whether inhibition of the proteolytic activity increases the susceptibility of F. hepatica immature worm to any extant immune effector mechanisms in hosts remains to be determined.

Animals