Routine analysis of amphetamine class drugs as their naphthaquinone derivatives in human urine by high-performance liquid chromatography

We describe a simple HPLC method which is suitable for the routine confirmation of immunoassay positive amphetamine urine samples. The precolumn derivisation method employing sodium naphthaquinone-4-sulphonate was found to have adequate sensitivity, selectivity and precision for the measurement of amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxyethylamphetamine (MDEA) at 500 microg/l cutoff level for confirmatory analysis of amphetamines in urine. The specificity of the method is enhanced by detecting the peaks at two different wavelengths. The ratios of the peak heights measured at the two wavelengths were different for each of the 5 amphetamines analysed. There was no interference from other phenylethylamine analogues that are commonly found in "over the counter" preparations. The HPLC method is compared to a commercial TLC system for detecting amphetamines in urine of drug abusers attending drug rehabilitation programmes. The HPLC confirmatory method described is a viable alternative to GC or to the more complex and costly GC-MS techniques for confirming amphetamine, methamphetamine, MDMA, MDA and MDEA in urine of drug abusers especially when used in a clinical care setting.     

Neuroactive substances and associated major muscle systems in<Emphasis Type="Italic"> Bucephaloides gracilescens</Emphasis> (Trematoda: Digenea) metacercaria and adult

Cholinergic, serotoninergic and neuropeptidergic components of the nervous system were examined and compared in the progenetic metacercaria and adult gasterostome trematode, Bucephaloides gracilescens in order to provide baseline information on neuronal control of the musculature involved in egg-assembly. Enzyme cytochemistry and indirect immunocytochemical techniques interfaced with confocal scanning laser microscopy demonstrated all three classes of neuroactive substance throughout the central and peripheral nervous systems. A comparable orthogonal arrangement of the central nervous system (CNS) and peripheral array of nerve plexuses was observed in both metacercaria and adult. Staining patterns for cholinergic and peptidergic substances showed significant overlap, while the serotoninergic system was confined to a separate set of neurons. Immunostaining for FMRFamide-related peptides (FaRPs) was strong in the CNS and peripheral innervation to the attachment apparatus of metacercaria and adult but was only found in the innervation of the ootype in ovigerous adults, implicating FaRPs in neuronal control of the muscle of the female reproductive tract during egg-assembly.     

The histological changes in transbronchial biopsy after treatment of acute lung rejection in heart-lung transplants

Transbronchial lung biopsies taken during episodes of acute lung rejection in heart-lung transplant patients were examined histologically. When the diagnosis was confirmed by microscopy, the patients were treated with augmented immunosuppression by high-dose corticosteroids. A second biopsy was obtained an average of 23.5 days after commencement of treatment. These biopsies were examined to determine the histological changes caused by treatment. In most cases, there were both quantitative and morphological differences between the infiltrates in acute rejection and in the remaining perivascular infiltrates after treatment. In acute rejection, 26 of 28 biopsies contained perivascular lymphocytic infiltrates, lymphocytes being large and blast-like. Although 20 of 28 follow-up biopsies still contained perivascular infiltrates, they were smaller and the lymphocytes smaller in size. Half the biopsies in rejection contained neutrophils, but less than half contained eosinophils in the perivascular infiltrates. After treatment, all these cells were less numerous. Another feature of treated rejection was the presence of haemosiderin around vessels suggesting earlier acute vascular injury. However, haemosiderin persists long after the cellular infiltrate has disappeared and cannot be considered a reliable feature of recently treated acute lung rejection. The bronchiolar infiltrates showed a similar pattern of responses to the perivascular infiltrates.     

An eighth complementation group of rodent cells hypersensitive to ultraviolet radiation

Two mutant lines (US31, US46) of mouse lymphoma cells that are hypersensitive to ultraviolet (UV) radiation were previously found to belong to different complementation groups. The mutants were tested for their ability to complement the six known complementation groups of UV-sensitive Chinese hamster ovary (CHO) cells, which are defective in nucleotide excision repair, as well as a seventh group represented by a V79 mutant. Hybrid cells were produced by fusion with polyethylene glycol and tested in situfor UV resistance. The mouse mutant US46 complemented all CHO mutants except UV61. Therefore, US46 is assigned to the same complementation group as UV61, and it is probably defective in the same locus. The mouse mutant US31 produced UV-resistant hybrid cells in each of the seven crosses, indicating that it forms an eighth complementation group among the rodent mutants. Thus, at least eight genes are likely required to repair UV damage in rodent cells.     

Sodium detection during the water absorption response in Rana pipiens

Although taste in vertebrates is typically associated with specialized receptors in the lingual epithelium, Hoff and Hillyard reported that the toad, Bufo punctatus, is able to "taste" sodium with the abdominal skin. This was reflected in a differential behavioral response to hypertonic NaCl. The present study tests for the presence of such abdominal chemoreceptors in the frog Rana pipiens. The experiment was a five-condition design in which frogs were placed on filter paper saturated with: deionized water, 250 mM NaCl, 350 mM NaCl, 12.9 microM amiloride, or 350 mM NaCl + 12.9 microM amiloride. The time that the frogs remained on the test substrate before moving to a surface of deionized water was recorded. It was necessary to dehydrate the frogs to 80% of their body weight to elicit a behavioral response to the NaCl whereas dehydration to 90% of their body weight has been reported effective in Bufo punctatus. The frogs displayed significantly shorter mean times to move on both concentrations of NaCl compared to deionized water, with the shortest times occurring when 350 mM NaCl was used. Amiloride alone did not have an effect upon times to move to deionized water, but did significantly reduce the response to 350 mM NaCl. Movement to amiloride + 350 mM NaCl did not differ significantly from that to deionized water. The results indicate that Rana pipiens, like Bufo punctatus, have epithelial chemoreceptors for the detection of NaCl on hydrated surfaces and that these receptors, like those of mammals, are amiloride sensitive.     

The effects of cycle racing on pulmonary diffusion capacity and left ventricular systolic function

The purpose of this study was to examine the effects of a 20 km cycle race (TT) on left ventricular (LV) systolic and pulmonary function in 12 endurance cyclists. Spirometry, single-breath diffusion capacity (DLCO) with partitioning of membrane (DM) and capillary blood volume (Vc) components and 2-D echocardiograms were performed before and after the TT. During the TT mean oxygen consumption was 3.79 +/- 0.5 L x min(-1) (83 +/- 5.5% of VO2max) and mean blood lactate was 8.4 +/- 2.4 mM. Following the TT, spirometry values were unchanged, however, DLCO and DM were significantly (P<0.05) reduced. LV systolic function was increased (P<0.05) immediately after exercise, while end-diastolic area was decreased (P<0.05) at all points during recovery. The reduction in DM was correlated with LV systolic function following the TT. This relationship suggests a cardiovascular contribution to pulmonary diffusion impairment following exercise.