Immuno-analysis of phospho-Akt in primary human breast cancers.

The aim of this study was to investigate the expression of the activated (phosphorylated) form of Akt (Ser473) in primary breast cancer and to correlate the results with clinicopathological and prognostic variables for clinically relevant associations. Phospho-Akt expression was studied using immunoblot analysis in 49 invasive breast carcinomas (median follow-up time 55 months, range 7-74 months). We assessed the level of phospho-Akt in different types of primary breast cancers and compared the use of autoradiograph X-ray film with a PVDF-DAB-staining system. Twelve percent of the tumours had no phospho-Akt protein, 25% had low phospho-Akt expression, 51% had intermediate expression and 12% had high phospho-Akt expression. No relationship was observed between phospho-Akt and tumour grade, tumour size or nodal status. A significant relationship was demonstrated between phospho-Akt score and oestrogen receptor status (P=0.014). Univariate analysis demonstrated that intermediate levels of phospho-Akt in breast tumour tissue are associated with a lower probability of developing recurrences (P=0.035), while in multivariate analyses, none of the phospho-Akt levels appeared to be independent predictors of disease recurrence or death. In addition, it has been clearly established that a suitable composition of reagents and components such as PVDF membranes treated with DAB substrate will enable the performing of sensitive immuno-analyses.

The impact on histopathology practice of new human tissue legislation in the UK.

The undisclosed or unauthorized retention of tissue from autopsies in the UK and elsewhere has caused considerable public concern and much distress to some families. Histopathologists involved in these cases have also been discomfited. These events have exposed deficiencies in prevailing legislation, principally in the Human Tissue Act 1961 and the Coroners Rules 1984. New human tissue legislation comes into force in the UK in September 2006. The Human Tissue Act 2004 and the Human Tissue (Scotland) Act 2006 make it unlawful to remove, store and use tissue from the dead without appropriate authority. The Human Tissue Act 2004, which does not apply in Scotland, also prohibits the removal, storage and use of tissue from living individuals for purposes specified in the Act unless appropriate consent has been obtained. The Coroners (Amendment) Rules 2005, which came into force in June 2005, introduced new arrangements for dealing with the retention of tissue from bodies undergoing coroner's autopsies. This new legislative regime is intended to create a climate in which pathologists, patients and the public can have confidence that tissue is used appropriately and, when necessary, with proper authority or valid consent. However, other than in Scotland, there may be unintended consequences arising from restrictions on archiving, for audit and diagnostic review, tissue samples from coronial autopsies.

Biological inferences from a mathematical model of comedo ductal carcinoma in situ of the breast.

The growth of a tumour in a duct is examined in order to model ductal carcinoma in situ (DCIS) of the breast, the earliest known stage of breast cancer. Interactions between the expansive forces created by tumour cell proliferation and the stresses that develop in the compliant basement membrane are studied using numerical and analytical techniques. Particular attention focuses on the impact that proteolytic enzymes have on the tumour's progression. As the tumour expands and the duct wall deforms, the tumour cells are subjected to mechanical and nutritional stresses caused by high pressures and oxygen deprivation. Such stresses may stimulate the cells to produce proteolytic enzymes that degrade the duct wall, making it more compliant and prone to penetration by the tumour cells. We use our model to compare these two hypotheses for enzyme production and find that mechanical stress is likely the dominant mechanism, with the wall deforming most at the centre of the duct. We then discuss the biological implications of our theoretical results and suggest possible directions for future work.

Mathematical modelling of comedo ductal carcinoma in situ of the breast.

The growth of a tumour in a cylindrical duct with compliant walls is examined in order to model the early stages of ductal carcinoma in situ (DCIS) of the breast, the earliest known stage of breast cancer. A nutrient-limited growth model is formulated, in which cell movement is described by a Stokes flow constitutive relation. The interactions between the expansive forces created by tumour cell proliferation and the stresses that develop in the compliant basement membrane are studied using asymptotic and numerical methods. In this way we show how the duct wall deforms as the tumour grows and also how the progression of the tumour along the duct depends upon the stiffness of the wall. By varying key parameters we determine how treatment, protease production and the inclusion of the surrounding stroma affect the growth. Finally, we discuss the biological relevance of our results and suggest possible directions for future work.

Modelling the early growth of ductal carcinoma in situ of the breast.

The growth of a tumour in a rigid walled cylindrical duct is examined in order to model the initial stages of tumour cell expansion in ductal carcinoma in situ (DCIS) of the breast. A nutrient-limited growth model is formulated, in which cell movement is described by a Stokes flow constitutive relation. The effects on the shape of the tumour boundary of the material properties (i.e. the viscosity) and the extent to which the cells adhere to the duct wall are studied using numerical and asymptotic methods. It is shown how stable, non-planar, interface configurations result and that, during these initial stages, before the duct wall has been breached, few cells die and a nutrient-rich model is usually sufficient to capture the behaviour. Finally, we discuss the relevance of this approach to DCIS and suggest possible avenues for further work.

Necropsy practice after the "organ retention scandal": requests, performance, and tissue retention.

AIMS: After the so called "organ retention scandal" in the UK this study set out to assess the impact on death certification and hospital (consent) necropsies, including the postmortem retention of tissues and organs. METHODS: Data were prospectively gathered over a one year period for all deaths occurring at the Royal Hallamshire Hospital, Sheffield, UK to determine the frequencies with which death certificates were completed and necropsies were requested. The seniority of the clinician undertaking these duties was recorded. Pathologists were asked to record the extent of every necropsy during the study period. The type and planned uses of tissues retained were recorded. RESULTS: Death certificates were issued for 88.5% of the 966 deaths for which clinicians completed proformas. Of these, 88.9% were issued by preregistration and senior house officers. Consent was sought for a necropsy in 6.2% of cases (usually by non-consultant staff) and was granted in 43.4% of these. The overall, medicolegal, and hospital necropsy rates were 13.4%, 9.9%, and 3.5%, respectively. Tissues were retained from 55.4% of necropsies for diagnostic purposes, although sampling does not appear to be systematic. CONCLUSIONS: Death certification and seeking consent for a necropsy are frequently delegated to junior clinical staff. This may explain the low standard of death certification reported by others and the low necropsy rate. The decline in the necropsy rate and the low rate of sampling for histological examination highlight the decline of the hospital necropsy and the lack of a systematic approach to tissue sampling.

Fibrinogen E fragment selectively disrupts the vasculature and inhibits the growth of tumours in a syngeneic murine model.

We recently demonstrated that a fragment of human fibrinogen, fibrinogen E fragment, inhibits the migration and differentiation of human endothelial cells in vitro. Here we show that it exerts similar effects on murine endothelial cells in vitro, and selectively disrupts tumour endothelium in vivo, causing widespread intravascular thrombosis and retarding the growth of CT26 tumours in a syngeneic murine model.

Primary endometrioid adenocarcinoma of the large intestine arising in colorectal endometriosis.

AIMS: Three cases of endometrioid adenocarcinoma arising in colorectal endometriosis are described with discussion of their macroscopic and microscopic pathology and diagnosis, using immunohistochemistry. METHODS AND RESULTS: Three middle-aged women presented with symptoms and signs of colorectal mass effect. Two had a preceding history of gynaecological endometriosis and all three had either been on hormone replacement therapy or had functioning ovaries prior to presentation with colorectal disease. Each underwent resection of tumours of the distal large intestine. The definitive diagnosis was dependent on histological examination and immunohistochemistry, which was used to demonstrate an origin in endometriotic tissue. CONCLUSIONS: Endometrioid adenocarcinoma is a rare complication of colorectal endometriosis, this report contributing to a total of 25 cases in the literature. Definitive diagnosis, aided by immunohistochemical studies, is important to enable the identification of the optimal management for this uncommon condition.