Bladder cancer: evaluation of staging accuracy using dynamic MRI.

AIM: To assess the accuracy of magnetic resonance imaging (MRI) in staging bladder cancer and to assess whether dynamic gadolinium-enhanced sequences have any added benefit in staging. MATERIALS AND METHODS: Over a 22 month period, the MRI findings of 100 consecutive patients with histologically proven transitional cell carcinoma (TCC) of the bladder were reviewed. The T stage was assessed independently on T2-weighted imaging alone and in combination with gadolinium-enhanced MRI. The final histological diagnosis was considered the reference standard. Statistical analysis was performed to ascertain stage-by-stage accuracy. Accuracy of MRI in differentiating superficial (≤ T1) from invasive (≥ T2) and in differentiating organ-confined (≤ T2) from non-organ-confined (≥ T3) disease was assessed. RESULTS: On a stage-by-stage basis, tumours were correctly staged using MRI in 63% of patients (observed agreement=0.63, weighted kappa=0.57). The sensitivity and specificity of MRI to differentiate between superficial (≤ T1) from invasive (≥ T2) disease was 78.2 and 93.3%. The observed agreement for this group was 85% (kappa=70%; p<0.0001). The sensitivity and specificity of MRI to differentiate between organ-confined (≤ T2) from non-organ confined (≥ T3) disease was 90.5 and 60%. The observed agreement for this group was 89% (kappa=30%; p<0.01). Gadolinium-enhanced images improved staging in only three patients. CONCLUSION: In the present study MRI was found to be a moderately accurate tool in assessing the T stage. Agreement on a stage-by-stage basis was good. Agreement for differentiating between non-invasive versus muscle-invasive disease was good and that for organ-confined versus non-organ-confined disease was fair. Routine use of gadolinium-enhanced images is not routinely required.

Renal oncocytoma: CT features cannot reliably distinguish oncocytoma from other renal neoplasms.

AIM: To retrospectively review the computed tomography (CT) imaging features of a series of histologically confirmed renal oncocytomas and to determine whether imaging features are predictive of this subtype of benign renal epithelial tumour. MATERIALS AND METHODS: From May 2001 to October 2007, 21 patients with 28 renal masses, confirmed as renal oncocytoma on histological examination of the resection specimen, were identified from the pathology database at our institution. The preoperative imaging findings were retrospectively analysed to determine characteristic features, if any, to predict this rare subtype of benign renal tumour. RESULTS: There were 11 female and 10 male patients and the age at presentation ranged from 40-80 years (mean age 65.9 years). The size of the masses ranged from 1.2-12 cm in diameter (mean diameter 4.9 cm). All masses showed contrast enhancement. In 18 (64.3%) lesions the enhancement of the tumour was isodense to renal cortex. Ten (35.7%) lesions were hypodense to renal cortex. In three (10.7%) lesions, a well-defined stellate central scar was seen at CT and confirmed pathologically. In two (7.1%) lesions, a central scar was identified pathologically, but not seen on CT. The size of the central scars ranged from 10-29 mm diameter on CT. Twenty-two (78.6%) lesions did not demonstrate a scar on CT or pathologically. None of the patients had regional lymphadenopathy or distant metastasis. CONCLUSION: Renal oncocytoma is typically described as being hypervascular and homogeneous, with a characteristic central stellate scar on CT. The present study demonstrates that these imaging features are found in only a small proportion of these tumours. Therefore, imaging characteristics alone are unreliable when differentiating between oncocytoma and renal cell carcinoma, and histopathological diagnosis remains the reference standard.

CD80-mediated induction of immunostimulation in two ocular melanoma cell lines is augmented by interferon-gamma.

Although the transfection of the T-cell costimulatory molecule CD80 cDNA into human tumours can augment their immunogenicity in vitro, its expression alone is ineffective in many tumour systems. We evaluated the influence of CD80 expression on the immunostimulatory activity of ocular melanoma cell lines and determined whether IFN-gamma could enhance the effect. Two ocular melanoma cell lines were transfected with CD80 cDNA. The immunostimulatory capacity of the CD80+ transfectants was determined by their ability to stimulate the proliferation of allogeneic peripheral blood mononuclear cells (PBMC). The influence of additional accessory molecules on PBMC proliferation was assessed by pre-treating the CD80 transfectants with IFN-gamma. The CD80+ transfectants induced proliferation of allogeneic PBMC. IFN-gamma treatment of the tumour cells induced upregulated expression of MHC class I, de novo expression of MHC class II and CD54, and enhanced the ability of the CD80+ transfectants to stimulate PBMC proliferation. CD4+ T cells were not required for the proliferative response against untreated CD80+ tumour cells but were necessary for the augmentation of proliferation observed following IFN-gamma treatment. CD80+ ocular melanoma cells possess immunostimulatory potential which is augmented by IFN-gamma induced upregulation of cell surface molecules. Further studies on the role of costimulatory molecules in inducing anti-tumour immunity in ocular melanoma may help to define new strategies for application of immunotherapeutic approaches to treat this aggressive disease.

Induction of human cytotoxic T lymphocytes that preferentially recognise tumour cells bearing a conformational p53 mutant.

The tumour-suppressor gene p53 is pivotal in the regulation of apoptosis, and point mutations within p53 are the commonest genetic alterations in human cancers. Cytotoxic T lymphocytes (CTL) recognise peptide-MHC complexes on the surface of tumour cells and bring about lysis. Therefore, p53-derived peptides are potential candidates for immunisation strategies designed to induce antitumour CTL in patients. Conformational changes in the p53 protein, generated as a result of point mutations, frequently expose the 240 epitope, RHSVV (amino acids 212-217), which may be processed differently from the wild-type protein resulting in an altered MHC-associated peptide repertoire recognised by tumour-specific CTL. In this study 42 peptides (37 overlapping nonameric peptides, from amino acids 193-237 and peptides 186-194, 187-197, 188-197, 263-272, 264-272, possessing binding motifs for HLA-A2) derived from the wild-type p53 protein sequence were assayed for their ability to stabilise HLA-A2 molecules in MHC class I stabilisation assays. Of the peptides tested, 24 stabilised HLA-A2 molecules with high affinity (fluorescence ratio >1.5) at 26 degrees C, and five (187-197, 193-200, 217-224, 263-272 and 264-272) also stabilised the complexes at 37 degrees C. Peptides 188-197, 196-203 and 217-225 have not previously been identified as binders of HLA-A2 molecules and, of these, peptide 217-225 stabilised HLA-A2 molecules with the highest fluorescence ratio. Peptide 217-225 was chosen to generate HLA-A2-restricted CTL in vitro; peptide 264-272 was used as a positive control. The two primary CTL thus generated (CTL-217 using peptide 217 225; and CTL-264 using peptide 264-272) were capable of specifically killing peptide-pulsed T2 or JY cells. In order to determine whether these peptides were endogenously processed and to test the hypothesis that mutants expressing different protein conformations would generate an alternative peptide repertoire at the cell surface, a panel of target cells was generated. HLA-A2+ SaOs-2 cells were transfected with p53 cDNA containing point mutations at either position 175 (R-->H) or 273 (R-->H) (SaOs-2/175 and SaOs-2/273). Two HLA-A2-negative cell lines, A431 and SKBr3, naturally expressing p53 mutations at positions 273 and 175 respectively, were transfected with a cDNA encoding HLA-A2. The results showed that primary CTL generated in response to both peptides were capable of killing SaOs-2/175 and SKBr3-A2 cells, which possess the same mutation, but not SaOs-2/273, A431-A2 or SKBr3 cells transfected with control vector. This suggests that these peptides are presented on the surface of SaOs-2/175 and SKBr3-A2 cells in a conformation-dependent manner and represent potentially useful target peptides for immunotherapy.

Infrequent expression of the MAGE gene family in uveal melanomas.

It has previously been reported that MAGE-1, -2, -3 and -4 genes are expressed in human cancers including cutaneous melanoma. MAGE-1 and MAGE-3 represent targets for specific immunotherapy because they encode peptide antigens which are recognised by cytotoxic T lymphocytes (CTL) when presented by HLA class I molecules, and pilot clinical trials with these peptides are currently in progress. It is likely that other members of the MAGE gene family may also encode antigens recognised by CTL. Uveal melanomas, like cutaneous melanomas, arise from melanocytes that are derived from the neural crest. To determine if uveal melanoma patients would be suitable for MAGE-peptide immunotherapy, the expression of MAGE-1, -2, -3 and -4 genes was assessed by reverse transcription followed by polymerase chain reaction (RT-PCR) amplification and ethidium bromide staining. Expression of MAGE genes was not detected in any of 27 primary tumours. Either MAGE-1 or MAGE-4 was expressed in only 2 of 26 metastatic samples, but expression of MAGE-2 or -3 was not detected. Our data suggest that, unlike cutaneous melanomas, uveal melanomas may not be suitable candidates for MAGE-peptide immunotherapy.

Identification of peptide epitopes of MAGE-1, -2, -3 that demonstrate HLA-A3-specific binding.

The MAGE gene family of tumour antigens are expressed in a wide variety of human cancers. We have identified 43 nonamer peptide sequences, from MAGE-1, -2 and -3 proteins that contain binding motifs for HLA-A3 MHC class I molecules. The T2 cell line, transfected with the cDNA for the HLA-A3 gene, was used in a MHC class I stabilisation assay performed at 37 degrees C and 26 degrees C. At 37 degrees C, 2 peptides were identified that stabilised HLA-A3 with high affinity (fluorescence ratio, FR > 1.5), 4 peptides with low affinity (FR 1.11-1.49) and 31 peptides that did not stabilise this HLA haplotype (FR < 1.1). At 26 degrees C, 12 peptides were identified that stabilised HLA-A3 with high affinity, 8 peptides with low affinity and 17 peptides that did not stabilise this HLA haplotype. Two peptides stabilised HLA-A3 at both temperatures. Small changes in one to three amino acids at positions distinct from the anchor residues altered peptide affinity. Data were compared to a similar study in which a peptide competition assay was used to investigate MAGE-1 peptide binding to several HLA haplotypes. This study demonstrates that anchor residues do not accurately predict peptide binding to specific HLA haplotypes, changes in one to three amino acids at positions distinct from anchor residues influence peptide binding and alternative methods of determining peptide binding yield different results. We are currently investigating the ability of these peptides to induce antitumour cytotoxic T lymphocyte activity as they may be of potential therapeutic value.

Power orientation: a study of female nurse and non-nurse managers.

Researchers studied the power orientations of female managers as related to career choice, education level, age, and years as a manager. The questionnaire includes six orientations to power: 1) power as good; 2) power as resource dependency; 3) power as instinctive drive; 4) power as political; 5) power as charisma; and 6) power as control and autonomy. Discerning power may explain how one perceives the managerial position.

The value of shoulder distension arthrography with intraarticular injection of steroid and local anaesthetic: a follow-up study.

A series of 51 patients, seen in Leicester, with "frozen shoulder", and referred for arthrographic examination, were assessed both before and after distension arthrography, using air and a low-osmolar contrast media combined with a steroid and local anaesthetic injection. 38 attended for further assessment at up to 6 months later. 16 patients were found to have a rotator cuff tear. There was no significant change in the mean range of active movement in the patients with rotator cuff tears, but symptomatic improvement ensued in 44% of cases. In those found to have no rotator cuff tear, and external rotation of less than 35 degrees, a significant improvement in range of movement was seen. While those with less limitation of external rotation showed no change in their range of movement, they did experience symptomatic improvement in 73% of cases. We conclude that shoulder distension arthrography, with steroid and local anaesthetic injection, may be of symptomatic benefit in patients with frozen shoulder and without a rotator cuff tear, while only those with external rotation of less than 35 degrees are likely to improve their range of motion.