Predicting the HER2 status of breast cancer from basic histopathology data: an analysis of 1500 breast cancers as part of the HER2000 International Study

The tests that are currently available for the measurement of overexpression of the human epidermal growth factor-2 (HER2) in breast cancer have shown considerable problems in accuracy and interlaboratory reproducibility. Although these problems are partly alleviated by the use of validated, standardised 'kits', there may be considerable cost involved in their use. Prior to testing it may therefore be an advantage to be able to predict from basic pathology data whether a cancer is likely to overexpress HER2. In this study, we have correlated pathology features of cancers with the frequency of HER2 overexpression assessed by immunohistochemistry (IHC) using HercepTest (Dako). In addition, fluorescence in situ hybridisation (FISH) has been used to re-test the equivocal cancers and interobserver variation in assessing HER2 overexpression has been examined by a slide circulation scheme. Of the 1536 cancers, 1144 (74.5%) did not overexpress HER2. Unequivocal overexpression (3+ by IHC) was seen in 186 cancers (12%) and an equivocal result (2+ by IHC) was seen in 206 cancers (13%). Of the 156 IHC 3+ cancers for which complete data was available, 149 (95.5%) were ductal NST and 152 (97%) were histological grade 2 or 3. Only 1 of 124 infiltrating lobular carcinomas (0.8%) showed HER2 overexpression. None of the 49 'special types' of carcinoma showed HER2 overexpression. Re-testing by FISH of a proportion of the IHC 2+ cancers showed that only 25 (23%) of those assessable exhibited HER2 gene amplification, but 46 of the 47 IHC 3+ cancers (98%) were confirmed as showing gene amplification. Circulating slides for the assessment of HER2 score showed a moderate level of agreement between pathologists (kappa 0.4). As a result of this study we would advocate consideration of a triage approach to HER-2 testing. Infiltrating lobular and special types of carcinoma may not need to be routinely tested at presentation nor may grade 1 NST carcinomas in which only 1.4% have been shown to overexpress HER2. Testing of these carcinomas may be performed when HER2 status is required to assist in therapeutic or other clinical/prognostic decision-making. The highest yield of HER2 overexpressing carcinomas is seen in the grade 3 NST subgroup in which 24% are positive by IHC. (C) 2003 Elsevier Science Ltd. All rights reserved.

Two-locus Linkage Analysis Applied to Putative Quantitative Trait Loci for Lipoprotein(a) Levels

Plasma levels of lipoprotein(a) _ Lp(a) _ are associated with cardiovascular risk (Danesh et al., 2000) and were long believed to be influenced by the LPA locus on chromosome 6q27 only. However, a recent report of Broeckel et al. (2002) suggested the presence of a second quantitative trait locus on chromosome 1 influencing Lp(a) levels. Using a two-locus model, we found no evidence for an additional Lp(a) locus on chromosome 1 in a linkage study among 483 dizygotic twin pairs.

Zygosity Diagnosis in the Absence of Genotypic Data: An Approach Using Latent Class Analysis

For zygosity diagnosis in the absence of genotypic data, or in the recruitment phase of a twin study where only single twins from same-sex pairs are being screened, or to provide a test for sample duplication leading to the false identification of a dizygotic pair as monozygotic, the appropriate analysis of respondents' answers to questions about zygosity is critical. Using data from a young adult Australian twin cohort (N = 2094 complete pairs and 519 singleton twins from same-sex pairs with complete responses to all zygosity items), we show that application of latent class analysis (LCA), fitting a 2-class model, yields results that show good concordance with traditional methods of zygosity diagnosis, but with certain important advantages. These include the ability, in many cases, to assign zygosity with specified probability on the basis of responses of a single informant (advantageous when one zygosity type is being oversampled); and the ability to quantify the probability of misassignment of zygosity, allowing prioritization of cases for genotyping as well as identification of cases of probable laboratory error. Out of 242 twins (from 121 like-sex pairs) where genotypic data were available for zygosity confirmation, only a single case was identified of incorrect zygosity assignment by the latent class algorithm. Zygosity assignment for that single case was identified by the LCA as uncertain (probability of being a monozygotic twin only 76%), and the co-twin's responses clearly identified the pair as dizygotic (probability of being dizygotic 100%). In the absence of genotypic data, or as a safeguard against sample duplication, application of LCA for zygosity assignment or confirmation is strongly recommended.

The blood volumes of the primary and secondary circulatory system in the Atlantic cod Gadus morhua L., using plasma bound Evans Blue and compartmental analysis

The volume of the primary (PCS) and secondary (SCS) circulatory system in the Atlantic cod Gadus morhua was determined using a modified dye dilution technique. Cod (N=10) were chronically cannulated in the second afferent branchial artery with PE-50 tubing. Evans Blue dye was bound to harvested fish plasma at a concentration of 1 mg dye ml(-1) plasma, and injected at a concentration of 1 mg kg(-1) body mass. Serial sampling from the cannula produced a dye dilution curve, which could be described by a double exponential decay equation. Curve analysis enabled the calculation of the primary circulatory and total distribution volume. The difference between these volumes is assumed to be the volume of the SCS. From the dilution curve, it was also possible to calculate flow rates between and within the systems. The results of these experiments suggest a plasma volume in the PCS of 3.42+/-0.89 ml 100 g(-1) body mass, and in the SCS of 1.68+/-0.35 ml 100 g(-1) body mass (mean +/- S.D.) or approximately 50% that of the PCS. Flow rates to the SCS were calculated as 2.7% of the resting cardiac output. There was an allometric relationship between body mass and blood volumes. Increasing condition factor showed a tendency towards smaller blood volumes of the PCS, expressed as percentage body mass, but this was not evident for the volume of the SCS.

The reliability of video based time motion analysis

Time motion analysis is extensively used to assess the demands of team sports. At present there is only limited information on the reliability of measurements using this analysis tool. The aim of this study was to establish the reliability of an individual observer's time motion analysis of rugby union. Ten elite level rugby players were individually tracked in Southern Hemisphere Super 12 matches using a digital video camera. The video footage was subsequently analysed by a single researcher on two occasions one month apart. The test-retest reliability was quantified as the typical error of measurement (TEM) and rated as either good (10% TEM). The total time spent in the individual movements of walking, jogging, striding, sprinting, static exertion and being stationary had moderate to poor reliability (5.8-11.1% TEM). The frequency of individual movements had good to poor reliability (4.3-13.6% TEM), while the mean duration of individual movements had moderate reliability (7.1-9.3% TEM). For the individual observer in the present investigation, time motion analysis was shown to be moderately reliable as an evaluation tool for examining the movement patterns of players in competitive rugby. These reliability values should be considered when assessing the movement patterns of rugby players within competition.

Applied physiology and game analysis of rugby union

Increased professionalism in rugby has elicited rapid changes in the fitness profile of elite players. Recent research, focusing on the physiological and anthropometrical characteristics of rugby players, and the demands of competition are reviewed. The paucity of research on contemporary elite rugby players is highlighted, along with the need for standardised testing protocols. Recent data reinforce the pronounced differences in the anthropometric and physical characteristics of the forwards and backs. Forwards are typically heavier, taller, and have a greater proportion of body fat than backs. These characteristics are changing, with forwards developing greater total mass and higher muscularity. The forwards demonstrate superior absolute aerobic and anaerobic power, and Muscular strength. Results favour the backs when body mass is taken into account. The scaling of results to body mass can be problematic and future investigations should present results using power function ratios. Recommended tests for elite players include body mass and skinfolds, vertical jump, speed, and the multi-stage shuttle run. Repeat sprint testing is a possible avenue for more specific evaluation of players. During competition, high-intensity efforts are often followed by periods of incomplete recovery. The total work over the duration of a game is lower in the backs compared with the forwards; forwards spend greater time in physical contact with the opposition while the backs spend more time in free running, allowing them to cover greater distances. The intense efforts undertaken by rugby players place considerable stress on anaerobic energy sources, while the aerobic system provides energy during repeated efforts and for recovery. Training should focus on repeated brief high-intensity efforts with short rest intervals to condition players to the demands of the game. Training for the forwards should emphasise the higher work rates of the game, while extended rest periods can be provided to the backs. Players should not only be prepared for the demands of competition, but also the stress of travel and extreme environmental conditions. The greater professionalism of rugby union has increased scientific research in the sport; however, there is scope for significant refinement of investigations on the physiological demands of the game, and sports-specific testing procedures.