Elevated expression of Ki-67 identifies aggressive prostate cancers but does not distinguish BRCA1 or BRCA2 mutation carriers.

Prostate cancers in men with germline BRCA1 and BRCA2 mutations are more aggressive than morphologically similar cancers in men without these mutations. This study was performed to test the hypothesis that enhanced expression of Ki-67, as a surrogate of cell proliferation, is a characteristic feature of prostate cancers occurring in BRCA1 or BRCA2 mutation carriers. The study cohort comprised 20 cases of prostate cancer in mutation carriers and 126 control sporadic prostate cancers. Of the combined sample cohort, 65.7% stained only within malignant tissues while 0.7% stained in both malignant and benign tissues (p<0.001). Significantly increased expression of Ki-67 occurred in prostate cancers with higher Gleason score (p<0.001). Elevated Ki-67 expression was identified in 71% of prostate cancers in BRCA1 or BRCA2 mutation carriers and in 67% of the sporadic controls (p>0.5). Similar results were obtained when the data were analysed using a threshold set at 3.5 and 7.1%. This study shows that elevated expression of Ki-67 is associated both with aggressive prostate cancers and with high Gleason score irrespective of whether their occurrence is against a background of BRCA1 or BRCA2 mutations or as sporadic disease. The data suggest that, since elevated Ki-67 does not distinguish prostate cancers occurring in BRCA1 or BRCA2 mutation carriers from sporadic prostatic malignancies, the effects of these genetic mutations are probably independent. While all prostate cancers occurring in the presence of BRCA germline mutations are clinically aggressive, their potentially different phenotypes consistently involve maximal rates of cell proliferation.

Transient hemiglossal denervation during acute internal capsule infarct in the setting of dysarthria-clumsy hand syndrome.

A case of MR imaging-documented transient unilateral tongue denervation presenting during acute internal capsule infarction is described. Understanding the corticolingual pathway innervation of the hypoglossal nucleus is essential for explaining these findings. Awareness of the findings in this case will facilitate appropriate diagnosis, provide neuroanatomic explanation, and prevent misdiagnosis.

Rapid development of post-radiotherapy sarcoma and breast cancer in a patient with a novel germline 'de-novo' TP53 mutation.

AIMS: Germline mutations in the TP53 tumour suppressor gene are associated with Li-Fraumeni syndrome, which is characterised by a spectrum of neoplasms occurring in children and young adults that predominantly include early-onset breast cancer, a variety of sarcomas, brain tumours and adrenocortical tumours. The identification of patients carrying TP53 mutations is primarily based on a positive family history of these early-onset characteristic cancer types. The aim of this study is to emphasize the importance of TP53 molecular testing in patients with very early onset breast cancer and no family history of cancer. MATERIALS AND METHODS: A young woman with no family history of cancer presented with bilateral breast cancer at the age of 27 years. Forty months later she developed malignant fibrous histiocytoma of the right clavicle and another primary left breast cancer. Molecular testing of mutations 185delAG, 5382insC in BRCA1 gene and 6174delT in BRCA2 gene was performed using multiplex PCR and separation on a denaturing polyacrylamide gel. TP53 molecular analysis was performed by PCR-SSCP analysis of the whole coding region of the TP53. Exon 8 PCR products were sequenced using an ABI dye terminator kit and examined on an ABI 3100 automated sequencer. RESULTS: Molecular testing of peripheral blood DNA did not reveal mutations in BRCA1 or BRCA2 genes. A novel germline TP53 mutation, c.G841C, p.D281N, was identified. The detected mutation is a missense substitution, c.G841C, resulting in the substitution of the amino acid aspartate to asparagine, p.D281N. Molecular analysis in her parents showed that neither of them carried the mutation. CONCLUSIONS: We describe a novel 'de novo'TP53 mutation and discuss the importance of molecular testing in early-onset breast cancer patients and its effect on the management and outcome of the disease.

Analysis by array CGH of genomic changes associated with the progression or relapse of Wilms' tumour.

Despite aggressive salvage regimens, approximately half of all children who suffer a Wilms' tumour recurrence will die of their disease. Although there are increasing data on molecular genetic prognostic factors present in the tumour at diagnosis, there is little information regarding the molecular events that occur with Wilms' tumour progression and relapse. In the present study, microarray-based comparative genomic hybridization (aCGH) analysis has been carried out on 58 Wilms' tumour samples, which included 38 untreated primary and 20 recurrent tumours. A higher degree of copy number changes was observed in the recurrent tumours (33.0% genomic clones) than in the primary tumour (21.2%). Paired analysis highlighted the acquisition of 15q gain with high levels of IGF1R expression in the tumour recurrence in two cases. The most statistically significant abnormality acquired between diagnosis and relapse was loss of 17p. One case that experienced 17p loss was classified as favourable histology at diagnosis, but exhibited diffuse anaplasia at recurrence and had a homozygous TP53 deletion. Another instructive case with a constitutional 11p13 deletion presented with bilateral tumours and suffered two subsequent recurrences in the left kidney. A somatic WT1 mutation was found only in the right kidney tumour, while the constitutional 11p13 deletion was the only abnormality detected in the initial left kidney tumour by aCGH. The two subsequent relapses in the left kidney contained an accumulation of additional genetic alterations, including an independent WT1 mutation.

CHEK2 variants in susceptibility to breast cancer and evidence of retention of the wild type allele in tumours.

We have recently shown that the CHEK2*1100delC mutation acts as a low penetrance breast cancer susceptibility allele. To investigate if other CHEK2 variants confer an increased risk of breast cancer, we have screened an affected individual with breast cancer from 68 breast cancer families. Five of these individuals were found to harbour germline variants in CHEK2. Three carried the 1100delC variant (4%). One of these three individuals also carried the missense variant, Arg180His. In the other two individuals, missense variants, Arg117Gly and Arg137Gln, were identified. These two missense variants reside within the Forkhead-associated domain of CHEK2, which is important for the function of the expressed protein. None of these missense variants were present in 300 healthy controls. Microdissected tumours with a germline mutation showed loss of the mutant allele suggesting a mechanism for tumorigenesis other than a loss of the wild type allele. This study provides further evidence that sequence variation in CHEK2 is associated with an increased risk of breast cancer, and implies that tumorigenesis in association with CHEK2 mutations does not involve loss of the wild type allele.

The value of rapid functional assays of germline p53 status in LFS and LFL families.

We have tested two rapid assays of p53 function, namely the apoptotic assay and the FASAY as means of detecting germline p53 mutations in members of Li-Fraumeni and Li-Fraumeni-like families. Results of the functional assays have been compared with direct sequencing of all 11 exons of the p53 gene. The results show good agreement between the two functional assays and between them and sequencing. No false-positives or negatives were seen with either functional assay although the apoptotic assay gave one borderline result for an individual without a mutation. As an initial screen the apoptotic assay is not only rapid but inexpensive and very simple to perform. It would be expected to detect any germline defect that leads to loss of p53 function. The apoptotic assay could be ideal as a means of prescreening large numbers of samples and identifying those that require further investigation. The FASAY detects mutations in exons 4-10, is rapid and distinguishes between functionally important and silent mutations.

Analysis of Li-Fraumeni syndrome and Li-Fraumeni-like families for germline mutations in Bcl10.

The Li-Fraumeni syndrome (LFS) is a dominant disease whose hallmark is an increased risk of breast cancers, brain tumours, sarcomas, leukaemia and adrenal carcinoma. Some, but not all LFS and Li-Fraumeni-like (LFL) families are caused by TP53 mutations. Bcl10 is a recently identified tumour suppressor reported to be commonly mutated in a wide range of cancers. To investigate the possibility that Bcl10 is a susceptibility gene for LFS and LFL we have analysed 27 LFS/LFL families. No mutations were observed. This indicates that Bcl10 is unlikely to act as a susceptibility gene for LFS and LFL.

On genetic and environmental factors in Menière's disease.

The etiology of Menière's disease (MD) remains obscure. Previous studies have shown a highly significant association between sporadic MD and one of the human leukocyte antigen, HLA-C genotypes, whereas disease activity has been related to the detection of enterovirus-specific viral protein (VP1) in the peripheral circulation. This present research extends the HLA association of sporadic cases to the study of families with more than one living member with unequivocal MD. Since the sporadic HLA associations point to chromosome 6 being a candidate region of a possible MD mutation, this area of the human genome has been investigated first; DNA suitable for study by other markers has been stored. The presence or absence of VP1 in the familial MD patients has been measured and related to disease activity at the time of sample collection. The association, in both sporadic and familial cases, of MD and partial HLA class I haplotypes points to a likely MD locus lying between the HLA-C and HLA-A loci on the short arm of chromosome 6. The significant relation between disease activity and circulating VP1 has been confirmed. It is likely that the predisposition to familial MD is attributable to a mutation on chromosome 6, which has been designated M1.

Calcium, magnesium and phosphorus status of elderly inpatients: dietary intake, metabolic balance studies and biochemical status.

The calcium, magnesium and phosphorus status of a group of elderly inpatients was studied by use of duplicate meal analysis over a 5 d period and biochemical indices in twenty-one patients, and metabolic balance (5 d) in six of these. Mean daily Ca intake was lower than that of apparently healthy elderly subjects in metabolic equilibrium, although commensurate with present UK recommendations. Metabolic balance was negative for Ca. Mean daily Mg intake was approximately half the US recommendation, and half the intake at which metabolic balance has been observed in healthy elderly people. The five patients studied were in metabolic balance for Mg. Mean daily P intake was close to the UK recommendation, but negative metabolic balance was observed. The disparity between official recommendations for Ca-intake, factors contributing to suboptimal Ca status, and measures that may improve Ca status in this group are discussed.

Iron status of hospitalized and housebound elderly people: dietary intake, metabolic balances, haematological and biochemical indices.

The iron status of housebound and hospitalized elderly people was studied by duplicate diet analysis, metabolic balance, biochemical and haematological investigations. Evidence of biochemical iron deficiency in the hospital group was accompanied by low dietary intake and negative metabolic balance. Metabolic equilibrium in the housebound group was seen with intakes similar to those found in healthy elderly though biochemical measures suggested a borderline iron deficient state.

Energy, protein, zinc and copper status of twenty-one elderly inpatients: analysed dietary intake and biochemical indices.

1. Duplicate diet analysis for energy, protein, zinc and copper with estimates of biochemical status for Zn and Cu were undertaken in twenty-one elderly long-stay inpatients (mean age 82 (range 63-89) years) consuming their customary hospital diet and in a stable medical condition. Fourteen patients had a long-standing and significant healing problem, either a leg ulcer or pressure sore. 2. Mean daily intakes of energy (5.2 MJ), protein (45 g), Zn (85 mumol) and Cu (14 mumol) were low in comparison with both official recommendations and levels of intake at which metabolic equilibrium was observed in healthy elderly people studied by the same methods (Bunker et al. 1984a). 3. Mean leucocyte Zn (9 pmol/10(6) cells) and Cu (7.5 pmol/10(6) cells) were low in comparison with results from healthy elderly people (Bunker et al. 1984a), implying suboptimal status for these elements. Those patients with healing problems tended to have the lower values within the range. 4. Recommendations are made with respect to improving nutritional status in this disadvantaged group of people.