This article reviews previously described methods of fresh tissue sampling from radical prostatectomy specimens for research and describes a method used in Oxford which is simple, logical and cost effective. The method utilises a systematic zonal approach to tissue procurement in order to meet the increasing requirement for research samples with detailed morphological information such as zone of origin, tumour stage and Gleason grade.The described method involves punch biopsy sampling from a 4mm thick transverse slice cut 8mm superior to the apex. 9 biopsies are taken from each specimen in the following zonal distribution: Mid anterior, right lateral, right peripheral zone lateral, right peripheral zone mid, left peripheral zone mid, left peripheral zone lateral, left lateral, left transition zone and right transition zone.The method was validated by successfully sampling tumour in 7 out of 8 cases (88%). In 6 of the positive cases, tumour was present in more than 1 punch biopsy. The mean time from receipt of the specimen to completion of the biopsy freezing was 23.5 minutes. Tumour stage, zone and Gleason grade were determined for all positive biopsies. All cases were reported to RCPath guidelines with no compromising of margins.A logical systematic method of fresh tissue sampling from radical prostatectomy specimens is presented, which balances the need for accurate routine histopathological reporting with the requirement for increasingly complex research samples to be taken with attention to morphological details such as zone and stage.
The role of focal brain damage as a trigger for autoimmune inflammation in multiple sclerosis (MS) is unclear. In this study we examine mechanisms by which experimental autoimmune encephalomyelitis (EAE) is enhanced by focal brain damage. EAE was produced in Lewis rats by footpad inoculation; focal brain damage, in the form of a cortical cryolesion (cryolesion-EAE), was induced 8 days post-inoculation (d.p.i.). The distribution of inflammation and chemokine production in cryolesion-EAE and EAE-only were compared. Inflammation in the brain, measured by immunocytochemistry for T lymphocytes (W3/13) and microglial activation (MHC class II -OX6), was significantly enhanced in cryolesion-EAE 11-15 d.p.i. (p < 0.01-0.05) but by 20-40 d.p.i., equated with EAE-only. Inflammation in cryolesion-EAE related to breakdown of the blood-brain barrier (BBB) at the site of the cryolesion and also to the corticospinal tracts and thalamus, reflecting the afferent and efferent neuronal connections with the cryolesioned cortex. Semiquantitative RT/PCR dot-blot hybridization assay showed a 6-fold increase in mRNA for specific chemokines in the brain in cryolesion-EAE at 9 d.p.i. (MCP-1) and 11 d.p.i. (MCP-1 and MCP-5) with no significant increase in RANTES, GRO-alpha, or MIP-1alpha. By 14 d.p.i., the levels of MCP-1 and MCP-5 mRNA equated with EAE-only animals. These results suggest that enhancement and location of autoimmune inflammation in the brain following focal cortical injury initially involve chemokines such as the macrophage chemoattractants MCP-1 and MCP-5, and the activities of afferent and efferent neuronal connections with the site of damage. By analogy, similar factors may modulate or reactivate autoimmune inflammation in MS.
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