Metabolism of 3-tert-butyl-4-hydroxyanisole to 3-tert-butyl-4,5-dihydroxyanisole by rat liver microsomes.

3-tert-Butylhydroxyanisole (3-BHA) is an antioxidant which can have a modulating effect on chemical carcinogenesis. Information concerning the metabolism of 3-BHA is incomplete. In the present study, the metabolites formed by incubating 3-BHA with liver microsomes from rats given beta-naphthoflavone by p.o. intubation were studied. Three metabolites were identified, two major metabolites and a minor metabolite. One of the major metabolites was the catechol of 3-BHA, i.e., 3-tert-butyl-4,5-dihydroxyanisole, which has not previously been reported. A characteristic of this compound is its capacity to be oxidized readily. The second major metabolite was tert-butyl hydroquinone which has been reported previously to be a liver microsomal metabolite of 3-BHA. The third metabolite, which occurred in small quantities, was 2,2'-dihydroxy-3,3'-di-tert-butyl-5,5'-dimethoxydiphenyl. 2,2'-Dihydroxy-3,3'-di-tert-butyl-5,5'-dimethoxydiphenyl has been identified previously as a major metabolite of 3-BHA in the rat intestine. An understanding of the metabolism of 3-BHA may assist in elucidating the mechanism(s) of its biological effects.

Performance of a new transdermal nitroglycerin adhesive patch formulation.

This two-period crossover study in 24 healthy men compared the transdermal absorption of nitroglycerin from a new 20-cm2 nitroglycerin adhesive transdermal patch applied for a single 24-hour period (q24hr) with that from one inch of 2% nitroglycerin ointment applied over a 50-cm2 area every eight hours (q8hr) during a single 24-hour period. The observed differences in pharmacokinetic parameters were expected, based on product design (q24hr vs q8hr) and surface area (20 cm2 vs 50 cm2); however, when corrected for surface area, the mean plasma AUC ratios indicated that the patch delivered about 1.5 times more drug than the ointment. The patch delivery of nitroglycerin was confirmed by patch residual results, which indicated nitroglycerin was released at a rate of 0.75 mg/cm2/24 hrs. The patch exhibited good skin adhesion throughout the 24-hour application period.

Comparison of the absorbed ELISA and agar gel immunodiffusion test with clinicopathological findings in ovine clinical paratuberculosis.

Thirty-two sheep with clinical paratuberculosis and 43 normal, healthy control animals were tested for serum antibodies to Mycobacterium avium subspecies paratuberculosis with the absorbed enzyme-linked immunosorbent assay (ELISA) and agar gel immunodiffusion (AGID) test. All the sheep were necropsied and the diseased cases were categorised as having either multibacillary ('lepromatous') or paucibacillary ('tuberculoid') intestinal lesions. The ELISA and AGID test were highly sensitive when testing the multibacillary group (86.4 per cent and 100 per cent, respectively) but the sensitivity of the tests in the paucibacillary group was significantly lower (10 to 50 per cent and 30 per cent, respectively). These findings were related to the ELISA optical density readings, with the multibacillary samples having values significantly greater than those of the paucibacillary and control groups, and the optical density values appearing to correlate with the numbers of mycobacteria present in the intestinal lesions. These results indicate both the usefulness of serological testing in the diagnosis of the multibacillary form of paratuberculosis and the difficulty in identifying animals with the paucibacillary form of the disease.

In vitro classical conditioning of abducens nerve discharge in turtles.

In vitro classical conditioning of abducens nerve activity was performed using an isolated turtle brainstem-cerebellum preparation by direct stimulation of the cranial nerves. Using a delayed training procedure, the in vitro preparation was presented with paired stimuli consisting of a 1 sec train stimulus applied to the auditory nerve (CS), which immediately preceded a single shock US applied to the trigeminal nerve. Conditioned and unconditioned responses were recorded in the ipsilateral abducens nerve. Acquisition exhibited a positive slope of conditioned responding in 60% of the preparations. Application of unpaired stimuli consisting of CS-alone, alternate CS and US, or backward conditioning failed to result in conditioning, or resulted in extinction of CRs. Latencies of CR onset were timed such that they occurred midway through the CS. Activity-dependent uptake of the dye sulforhodamine was used to examine the spatial distribution of neurons labeled during conditioning. These data showed label in the cerebellum and red nucleus during conditioning whereas these regions failed to label during unconditioned responses. Furthermore, the principal abducens nucleus labeled heavily during conditioning. These findings suggest the feasibility of examining classical conditioning in a vertebrate in vitro brainstem-cerebellum preparation. It is postulated that the abducens nerve CR represents a behavioral correlate of a blink-related eye movement. Multiple sites of conditioning are hypothesized, including the cerebellorubral circuitry and brainstem pathways that activate the principal abducens nucleus.

Changes in perceived contrast of suprathreshold gratings as a function of orientation and spatial frequency.

Orientation anisotropy for suprathreshold gratings of different spatial frequencies was measured using a contrast matching procedure. Observers matched the contrast of sine-wave gratings of various orientations to a vertical reference grating set at different reference contrasts. At threshold, the size of the anisotropy increased with spatial frequency, confirming previous results. When the reference grating contrast was set above threshold, the anisotropy declined, and eventually disappeared for gratings of medium spatial frequencies. At higher spatial frequencies, although the relative anisotropy became smaller, it did not disappear within the range of contrasts used in this study. For medium, but not for high spatial frequencies, the data are consistent with Kulikowski's (1976) model of effective contrast constancy.

Effect of food on salsalate absorption.

To assess the effect of food on salsalate absorption, single 1500-mg oral doses of salsalate were administered to 17 men under fasted and fed conditions according to a randomized open-label crossover design. A 7-day washout separated treatment periods. Blood samples were drawn throughout the 48-h period following dose administration and the resulting plasma samples assayed by high-performance liquid chromatography (HPLC) for unchanged drug, salsalate, and the major metabolite, salicylic acid. When results for the fasted and fed treatments were compared, no significant differences were observed in the pharmacokinetic parameters for the major metabolite salicylic acid or in the extent of absorption of unchanged drug; however, the rate of salsalate absorption was affected. Although the time-to-peak for salsalate was significantly delayed by approximately 1 h in the presence of food, the peak level was not significantly affected. The lack of difference between the two treatments for the therapeutic moiety, salicylic acid, indicates a lack of a significant food effect on single doses of salsalate.