Confocal laser endomicroscopy in experimental pigs. Methods of ex vivo imaging.

BACKGROUND: Confocal laser scanning endomicroscopy (CLSE) enables online in vivo cellular surface and subsurface imaging of normal and pathological tissue at high resolution and magnification. The aim of this study was to work out a method of ex vivo in vitro CLSE in experimental pigs and to compare CLSE images with those of "classic" histology. MATERIAL AND METHODS: Five mature female pigs entered the study. CLSE on an ex vivo in vitro basis was started 10 minutes after pharmacological euthanasia and carried out for 30 minutes. Fluorescein was administrated i.v. as a fluorescence substance. RESULTS: CLSE was successful in all tissue samples of all animals (the oesophagus, stomach, small intestine, large bowel). We have succeeded to obtain high quality images within the first 30 minutes that means 40 minutes after the euthanasia of experimental animals. CLSE images corresponded well with those of haematoxylin-eosin staining. CONCLUSIONS: CLSE on an ex vivo in vitro basis in experimental pigs is feasible.

Use of electrogastrography in preclinical studies of cholinergic and anticholinergic agents in experimental pigs.

Electrogastrography (EGG) is a non-invasive method for the assessment of gastric myoelectrical activity. Porcine EGG is comparable with human one. The purpose of this study was to evaluate the effect of atropine and neostigmine on the EGG in experimental pigs. Adult female pigs were administrated atropine (1.5 mg i.m., n=6) and neostigmine (0.5 mg i.m., n=6) after the baseline EGG, followed by a 90-min trial recording (MMS, Enschede, the Netherlands). Running spectral analysis was used for the evaluation. The results were expressed as dominant frequency of slow waves and EGG power (areas of amplitudes). Neostigmine increased continuously the dominant frequency and decreased significantly the EGG power. Atropine did not change the dominant frequency significantly. However, atropine increased significantly the EGG power (areas of amplitudes) from basal values to the maximum at the 10-20-min interval. After that period, the areas of amplitudes decreased significantly to the lowest values at the 60-90-min interval. In conclusion, cholinergic and anticholinergic agents affect differently EGG in experimental pigs.

Identification and determination of phase II nabumetone metabolites by high-performance liquid chromatography with photodiode array and mass spectrometric detection.

Chromatographic analyses play an important role in the identification and determination of phase I and phase II drug metabolites. While the chemical standards of phase I metabolites are usually available from commercial sources or by various synthetic, degradation or isolation methods, the phase II drug metabolites have usually more complicated structures, their standards are in general inaccessible and their identification and determination require a comprehensive analytical approach involving the use of xenobiochemical methods and the employment of hyphenated analytical techniques. In this work, various high-performance liquid chromatography (HPLC) methods were employed in the evaluation of xenobiochemical experiments leading to the identification and determination of phase II nabumetone metabolites. Optimal conditions for the quantitative enzymatic deconjugation of phase II metabolites were found for the samples of minipig bile, small intestine contents and urine. Comparative HPLC analyses of the samples of above-mentioned biomatrices and of the same biomatrices after their enzymatic treatment using beta-glucuronidase and arylsulfatase afforded the qualitative and quantitative information about phase II nabumetone metabolites. Hereby, three principal phase II nabumetone metabolites (ether glucuronides) were discovered in minipig's body fluids and their structures were confirmed using liquid chromatography (LC)-electrospray ionization mass spectrometric (MS) analyses.

Morphometric analysis of the porcine gastrointestinal tract in a 10-day high-dose indomethacin administration with or without probiotic bacteria Escherichia coli Nissle 1917.

BACKGROUND: Nonsteroidal anti-inflammatory drugs may cause severe injury to all parts of the gastrointestinal tract. It has been hypothesised that probiotic bacteria might reduce this adverse effect. The aim of this study was to perform a morphometric evaluation of the gastrointestinal tract in experimental pigs treated using a 10-day high-dose of indomethacin with or without Escherichia coli Nissle 1917 (EcN). METHODS: Twenty-four healthy mature pigs were included: Group A (controls; 6 animals), Group B (EcN; n = 6), Group C (indomethacin; n = 6) and Group D (EcN & indomethacin; n = 6). EcN (3.5 × 10(10) live bacteria/day for 14 days) and/or indomethacin (15 mg/kg/day for 10 days) were administered. Specimens of the stomach, small and large bowel were routinely processed for microscopic examination. The height of glandular mucosa, height and width of interfoveolar spaces and villi and basement size of epithelial cells were evaluated. RESULTS: Different effects of indomethacin and EcN on particular parts of the gastrointestinal tract were shown. The indomethacin and probiotics group demonstrated a significantly lower height of cryptal mucosa and colonocytes and widening of the basement size of colonocytes compared to controls (p = 0.004; p < 0.001; p = 0.025). The height of cryptal mucosa was significantly higher in the EcN group compared to controls (p = 0.001). CONCLUSIONS: Indomethacin alone induced marked adaptation of the gastric mucosa. EcN alone provided a significant favourable trophic effect on the colonic mucosa. However, indomethacin and probiotics administered together comprise the worst impact on all porcine stomach, small and large bowel.