Compartmental resection of peripheral nerve tumours with limb preservation in 16 dogs (1995-2011).

Peripheral nerve tumours (PNTs) affecting the limbs may lead to chronic pain, lameness and/or monoparesis that is refractory to medical treatment. The most common radical therapy for PNTs has been surgical excision with limb amputation. However, compartmental resection with preservation of the limb has been performed by the authors with favourable clinical results and therefore this bi-institutional retrospective study was undertaken to assess limb function, survival and recurrence. Sixteen dogs that had been diagnosed with PNTs between 1995 and 2011 met the inclusion criteria for this study. In the majority of the cases, good to excellent limb function was achieved. The overall median survival time (MST) was 1303days (42.8 months; range, 14 days-4639 days, [0.5-152.4 months]), with two dogs still alive at time of evaluation. Non-infiltrated margins were the best prognostic indicator; dogs with non-infiltrated margins had a MST of 2227days (P<0.001) compared to dogs with infiltrated margins (MST of 487 days). The 1-year calculated survival rate was 68.8% and the 2- and 3-year calculated survival rates were 62.5%. Surgical treatment with tumour removal and limb spare for proximal and distal PNTs can be successful. Compartmental excision can lead to good limb function, producing survival comparable to limb amputation, and should therefore be considered as an alternative to limb amputation in canine PNTs.

Developments in the application of gas chromatography with atomic emission (plus mass spectrometric) detection.

Capillary gas chromatography with atomic emission detection is a highly element-selective and sensitive detection technique for many non-metal as well as metallic elements. A 3-5 order of magnitude element/carbon selectivity, compound-independent calibration and the possibility to calculate (partial) molecular formulae are some of the attractive features of the technique. In the present review, the emphasis is on real-life applications for non-metals such as sulphur, phosphorus, nitrogen and the halogens, and on the potential of combined atomic emission/mass spectrometric detection.

Attempt to unravel the composition of toxaphene by comprehensive two-dimensional gas chromatography with selective detection.

Comprehensive two-dimensional gas chromatography (GC x GC) coupled with micro electron-capture and time-of-flight mass spectrometric (TOF-MS) detection has been used to analyse technical toxaphene. An HP-1 x HT-8 column combination yielded highly structured chromatograms and revealed a complex mixture of over 1000 compounds what is significantly higher number than in any study before. The analysis of a mixture of 23 individual congeners and TOF-MS evaluation of technical toxaphene showed that the chromatogram is structured according to the number of chlorine substituents in a molecule. The nature of the compounds (bornane and camphene) does not appear to have any influence. The sum of the peak areas of all congeners in each group was calculated using laboratory-written software; based on these results, the composition of technical toxaphene as a function of the number of chlorine substituents was provisionally calculated and was found that hepta- and octachlorinated compounds represents 75% of the total toxaphene area.

Cytotoxicity of paracetamol and 3,5-dihalogenated analogues: Role of cytochrome P-450 and formation of GSH conjugates and protein adducts.

The effect of 3,5-dihalogenation of paracetamol (PAR) on the cytotoxicity in rat hepatocytes isolated from beta-naphthoflavone pretreated, non-fasted rats, and the role of cytochrome P-450 in this regard, were studied. On incubation, 3,5-difluoro-PAR, 3,5-dichloro-PAR and 3,5-dibromo-PAR, as well as PAR, caused severe leakage of lactate dehydrogenase (LDH) which was preceded by a rapid concentration- and time-dependent depletion of intracellular glutathione (GSH). IC(50) values, representing the concentration of compound that caused 50% GSH depletion after 30 min of incubation, varied from 0.1 to 0.5 mM. This LDH leakage and GSH depletion could be inhibited by 1-ethynylpyrene. In hepatocytes from uninduced rats, GSH depletion was much less prominent and the concomitant LDH leakage almost completely absent. HPLC analysis of soluble metabolites and gas chromatography-mass spectrometry analysis, after alkaline peralkylation of the protein fraction, revealed (a) that 3,5-dihalogenated PAR analogues were liable to structure-related detoxification by glucuronidation, and (b) analogous to PAR, a substantial amount of each 3,5-dihalogenated PAR analogue was bioactivated by cytochrome P-450, ultimately leading to GSH-conjugates as well as (for 3,5-dichloro-PAR and 3,5-dibromo-PAR), protein adducts at regio-specific aromatic positions.

Rat liver microsomal cytochrome P450-dependent oxidation of 3,5-disubstituted analogues of paracetamol.

1. The cytochrome P450-dependent binding of paracetamol and a series of 3,5-disubstituted paracetamol analogues (R = -F, -Cl, -Br, -I, -CH3, -C2H5, -iC3H7) have been determined with beta-naphthoflavone (beta NF)-induced rat liver microsomes and produced reverse type I spectral changes. Ks,app varied from 0.14 mM for 3,5-diiC3H7-paracetamol to 2.8 mM for paracetamol. 2. All seven analogues underwent rat liver microsomal cytochrome P450-dependent oxidation, as reflected by the formation of GSSG in the presence of GSH. The GSSG-formation was increased in all cases upon pretreatment of rats by beta-naphthoflavone (beta NF) and was generally decreased upon pretreatment by phenobarbital (PB). 3. Rat liver microsomal cytochrome P450 as well as horseradish peroxidase catalysed the formation of 3,5-disubstituted NAPQI analogues from the corresponding parent compounds, as identified by UV-spectrophotometry of the NAPQI analogues and by GC/MS detection of the following GSH-conjugates: 2-glutathione-S-yl-3,5-dimethyl-1,4-dihydroxybenzene, 2-glutathione-S-yl-3,5-dichloro-paracetamol, and 2-glutathione-S-yl-3,5-dibromo-paracetamol. 4. In liver microsomal (beta NF-induced) incubations, apparent K(m) values, as determined for the cytochrome P450 catalysis-dependent oxidation of GSH, for seven 3,5-disubstituted paracetamol analogues (R = -F, -Cl, -Br, -I, -CH3, -C2H5, iC3H7) varied from 0.07 to 0.64 mM. Paracetamol exhibited an apparent K(m) of 0.73 mM. Apparent Vmax values for the cytochrome P450 catalysis dependent oxidation of GSH varied from 0.66 nmol min-1 mg-1 protein for paracetamol to 3.0 nmol min-1 mg-1 protein for 3,5-dimethyl-paracetamol.