HER2 amplification status in breast cancer: a comparison between immunohistochemical staining and fluorescence in situ hybridisation using manual and automated quantitative image analysis scoring techniques.

AIMS: To compare the results of breast cancer sections with HercepTesttrade mark immunohistochemistry (IHC) scores ranging from 0 to 3+ with fluorescence in situ hybridisation (FISH) for HER2 amplification. The HER2 digital scoring application of the Micrometastasis Detection System (MDS) was used, together with manual scoring of FISH and HercepTest, to determine whether this system provides an accurate alternative. METHODS: Paraffin wax embedded sections were stained using HercepTest and analysed by eye and automated quantitative image analysis. FISH was performed using the PathVysion fluorescent probe and scored by eye and automated quantitative image analysis using MDS. RESULTS: Of 114 cases, 26% were amplified by FISH, whereas only 18% scored 3+; 32% of IHC 2+ cases were amplified by FISH, and one showed borderline amplification. Six percent of IHC negative cases (0 or 1+) were amplified by FISH, and one showed borderline amplification. Of IHC 3+ cases, 10% were non-amplified by FISH. Classification discrepancies were seen in 18% of HercepTest cases scored by eye and using the MDS system. MDS was consistent with visual FISH scoring and correctly differentiated most ambiguous visual IHC scores. CONCLUSIONS: FISH provides a more accurate and consistent scoring system for determining HER2 amplification than HercepTest. The MDS system provides a reliable, consistent alternative to visual IHC and FISH scoring. IHC is still a valuable technique to aid in identification of isolated or heterogeneous tumour populations for subsequent FISH analysis, and a combined FISH and HercepTest approach to all breast cancer cases may be the most efficient strategy.

Chylous ascites: a manifestation of blunt abdominal trauma in an infant.

This report presents an infant in whom a unique case of chylous ascites developed after blunt abdominal trauma. Unfortunately, this case was complicated by Pseudomonas peritonitis, likely from a distant source. Our patient was treated medically and had a good overall outcome.

Detergent sensitivity of the tonoplast H(+)-ATPase and its purification from Beta vulgaris.

Tonoplast membrane fractions were isolated from beetroot (Beta vulgaris) using a refined method of preparation which significantly improved the yield of active tonoplast H(+)-ATPase, and electron microscopy showed these fractions to be a preparation of small vesicles, of diameter 500 nm to 50 nm and a minor fraction consisting of mainly tubular membrane structures of diameter 5 nm and length up to 1 micron. The stability of the tonoplast H(+)-ATPase was assessed in the presence of many biological detergents, using a linked assay. The addition of detergent to tonoplast membranes generally led to an increase in ATPase activity, and activity was maintained in a wide range of both non-ionic and zwitterionic detergents. Using the non-ionic detergent dodecyl maltoside, the tonoplast H(+)-ATPase was partially purified using ion-exchange chromatography on an HPLC system. Very high rates of ATP hydrolysis were recorded in these fractions. The purified membranes behaved as expected in the presence of known activators and inhibitors. An unexpected observation, however, was that low concentrations of vanadate could significantly increase the rate of H(+)-ATPase activity.

The evolution of acute renal failure, 1956-1988.

A total of 1347 patients with severe acute renal failure, treated at a single centre between 1956 and 1988, are reviewed. Only patients with an acute uraemic episode requiring dialysis and/or with serum creatinine levels above 600 mumols/l were included. The age of patients increased from median 41.25 years in the 1950s to 60.5 years in the period 1980 to 1988. The case-mix also altered with a decline in obstetric and traumatic disease, both of which carried an excellent prognosis, and an increase in the number of elderly patients with complicated medical and surgical conditions. Survival significantly decreased with increasing age and in the presence of complicating factors such as sepsis or malignancy. Despite these changes, there has been a progressive improvement in survival from 48.8 per cent in 1956-1959 to 57.9 per cent in 1985-1988. Survival for medical and surgical cases has improved from 38.5 per cent in 1956-1959 to 57.9 per cent in 1985-1988 (overall survival for non-obstetric cases was 44.2 per cent), due to improved prognosis for acute renal failure in general and acute renal failure due to intrinsic renal disease in particular.