A new guinea-pig model for acute inhalation studies on combined effects of cold air and pollutants.

We have developed a new small animal model for acute inhalation studies on combined effects of cold air and gaseous urban air pollutants. The anaesthetised, tracheostomised and paralysed guinea-pig was placed inside a small, sealed whole-body-box, in which it was ventilated mechanically by using cyclic negative pressure (Pbox) for active expansion of the chest. During a 2-h normal ventilation with warm humid air (n=6), there was a need for increasing Pbox with time to maintain the fixed tidal volume (VT) of 11 ml/kg. No such need was seen in the experiments with 15-min periods of isocapnic hyperventilation at 80 and 120 breaths/min (n=13). During the 2-h normal ventilation and in experiments with hyperventilation, there was a gradual increase in heart rate and small gradual decreases in PaCO2 and pH with time. Cold air + SO2 2.5 ppm produced a significantly stronger bronchoconstriction (deltaVT=-30.3+/-7.2%, n=6, P < 0.05) than clean cold dry air (deltaVT=-10.6+/-1.3%, n=6) and cold air + NO2 2.5 ppm (deltaVT=-13.2+/-3.3%, n=6), although these three exposure conditions produced similar decreases in tracheal air and retrotracheal tissue temperatures. With the present guinea-pig model, the combined respiratory effects of cold air and gaseous urban air pollutants can be investigated in a highly controlled manner.

Nerve function and its determinants in patients with newly-diagnosed type 2 (non-insulin-dependent) diabetes mellitus and in control subjects--a 5-year follow-up.

In order to assess the changes in nerve function 5 years after the diagnosis of diabetes mellitus and the determinants of progression of neuropathy, we studied 113 Type 2 (non-insulin-dependent) diabetic patients and 127 non-diabetic control subjects. Motor and sensory nerve conduction velocities were measured at the time of diagnosis of diabetes and 5 years later. At both examinations conduction velocities and response amplitudes were lower in diabetic patients than in control subjects. During the follow-up sural nerve conduction was impaired in both diabetic and control subjects, but, in general, changes in neurophysiological parameters were slight and inconsistent. In 12 diabetic patients nerve function deteriorated significantly during the follow-up. These patients had higher glycaemic indices at both examinations and lower baseline blood pressure levels as compared to the rest of the diabetic patients. No differences between these patient groups were found in other baseline risk factors (age, obesity, use of alcohol, smoking, serum insulin levels, albuminuria, lipids). In conclusion, Type 2 diabetic patients have disturbed nerve function at the time of diagnosis, but neurophysiological impairment during the next 5 years is on the average slight. Poor glycaemic control seems to be the most important risk factor in the deterioration of nerve function in these patients.

Studies on the chromatographic fractionation of metabolites of benzo[a]pyrene in faeces and urine from germfree and conventional rats.

Rats, germfree and conventional, were dosed with 14C-labelled benzo[a]pyrene. Faeces and urine were collected. Metabolites in faeces were effectively extracted with a new method using a combination of solvents and solid sorbents. Metabolites in urine were extracted with octadecylsilane-bonded silica. The metabolites were fractionated into groups by chromatography on a cation exchanger (SP-LH-20 or SP-Sephadex C-25) and an anion exchanger (TEAP-LH-20). Some of the groups were further purified by column chromatography and analysed by HPLC and TLC. The analyses show a complex pattern of metabolism. A large part of the metabolites (9-24% depending on animal type and route of excretion) had amphoteric properties, e.g. like glutathione and cysteine conjugates. The abundance of conjugates sensitive to beta-glucuronidase and sulphatase was low. The relative amount of acidic conjugates in faeces was much higher in the germfree than in the conventional rats indicating the influence of the intestinal flora on the metabolism. The results support the view that the mercapturic acid pathway is a quantitatively important metabolic route for benzo[a]pyrene in rats. The methods of extraction and group fractionation were designed to be generally applicable to the analysis of lipophilic xenobiotics and their metabolites.

Decreased GABA, benzodiazepine, and picrotoxinin receptor binding in brains of rats after cobalt-induced epilepsy.

In previous studies of experimental and human epilepsy, defects have been shown in the gamma-aminobutyric acid (GABA) receptors. We further investigated the role of the GABA/benzodiazepine/picrotoxinin receptor complex in the epileptic focus and also in other regions of the rat brain. The focus was induced by cobalt implantation to the right motor cortex, and the brains were dissected 16-19 days after the operation. Benzodiazepine (using [3H]flunitrazepam as a ligand; FLU), GABA [3H]muscimol; MUS), and picrotoxinin [( 35S]t-butylbicyclophosphorothionate; TBPS) receptor bindings were measured in different brain areas and the values were compared with glass-implanted controls. In the focal area, the specific receptor binding decreased in the order TBPS greater than FLU greater than MUS. In the perifocal area only TBPS binding decreased, and Scatchard analysis showed a decrease in the number of binding sites (p less than 0.05) without any effect on binding affinity. No change was seen in the binding characteristics of the other areas studied. According to our results, in cobalt-induced epilepsy the GABA/benzodiazepine/picrotoxinin receptor complex is modulated in the focal area; this may lead to a defect in chloride conductance, which in turn induces disturbed control of neuronal activity in the epileptic focus.