Search

Familial breast cancer has been recognised for many years. In the 1990s the genetic mechanism of inheritance of a proportion of these familial cancers was found to be attributable to germline mutation in either of two newly discovered genes, namely BRCA1 and BRCA2. Since the discovery of these genes, studies have been performed in which the pathological characteristics of familial cancers arising in patients with germline BRCA1 and BRCA2 mutation have been examined. A distinct pathological phenotype of high-grade, oestrogen receptor-negative breast cancer, often with medullary features, has been consistently described for BRCA1 cancers. A less distinct phenotype has been described for BRCA2 cancers. The discovery of genotype-phenotype correlation has significant implications for patient management and novel treatment strategies, not only for inherited cancers, but for breast cancer in general.

Diffuse lung disease (DLD) of infancy has multiple aetiologies and the spectrum of disease is substantially different from that seen in older children and adults. In many cases, a specific diagnosis renders a dire prognosis for the infant, with profound management implications. Two recently published series of DLD of infancy, collated from the archives of specialist centres, indicate that the majority of their cases were referred, implying that the majority of biopsies taken for DLD of infancy are first received by less experienced pathologists. The current literature describing DLD of infancy takes a predominantly aetiological approach to classification. We present an algorithmic, histological, pattern-based approach to diagnosis of DLD of infancy, which, with the aid of appropriate multidisciplinary input, including clinical and radiological expertise and ancillary diagnostic studies, may lead to an accurate and useful interim report, with timely exclusion of inappropriate diagnoses. Subsequent referral to a specialist centre for confirmatory diagnosis will be dependent on the individual case and the decision of the multidisciplinary team.

Purpose Endometrial adenocarcinoma (EC) is the most common gynaecologic cancer. Up to 90% of EC patients are obese which poses a health threat to patients post-treatment. Standard treatment for EC includes hysterectomy, although this has significant side effects for obese women at high risk of surgical complications and for women of childbearing age. This trial investigates the effectiveness of non-surgical or conservative treatment options for obese women with early stage EC. The primary aim is to determine the efficacy of: levonorgestrel intrauterine device (LNG-IUD); with or without metformin (an antidiabetic drug); and with or without a weight loss intervention to achieve a pathological complete response (pCR) in EC at six months from study treatment initiation. The secondary aim is to enhance understanding of the molecular processes and to predict a treatment response by investigating EC biomarkers. Methods An open label, three-armed, randomised, phase-II, multi-centre trial of LNG-IUD ± metformin ± weight loss intervention. 165 participants from 28 centres are randomly assigned in a 3:3:5 ratio to the treatment arms. Clinical, quality of life and health behavioural data will be collected at baseline, six weeks, three and six months. EC biomarkers will be assessed at baseline, three and six months. Conclusions There is limited prospective evidence for conservative treatment for EC. Trial results could benefit patients and reduce health system costs through a reduction in hospitalisations and through lower incidence of adverse events currently observed with standard treatment.

BACKGROUND Women with breast carcinoma diagnosed before age 40 years have a greater prevalence of germline BRCA1 and BRCA2 mutations than women with breast carcinoma diagnosed at older ages. Several recognizable histologic characteristics have been identified in breast carcinoma from studies of BRCA1/2 mutation carriers who belong to multiple-case families. The authors attempted to determine whether breast carcinoma occurring before age 40 years in BRCA1 or BRCA2 mutation carriers who were not selected for family history could be distinguished histologically from one another and from breast carcinoma in women of a similar age without a germline BRCA1 or BRCA2 mutation. METHODS The study undertook a histologic assessment of breast carcinomas diagnosed before age 40 years identified from a population-based study. RESULTS Breast carcinoma in BRCA1 mutation carriers was associated with a distinct histologic appearance; these tumors were high grade, and had exceptionally high mean mitotic counts, a syncytial growth pattern, pushing margins, and confluent necrosis. Atypical medullary carcinoma was overrepresented in BRCA1 mutation carriers. All low grade tumors and tumors with low mitotic rates belonged to the group without BRCA1 or BRCA2 mutations. Pleomorphic lobular carcinomas and extensive intraduct carcinomas were more common in BRCA2 mutation carriers. CONCLUSIONS Breast carcinoma occurring in women with a germline BRCA1 or BRCA2 mutation have recognizable histologic phenotypes, which may be useful in identifying individuals more likely to carry germline mutations. Histologic examination of breast carcinoma should become an important part of the evaluation of women seeking genetic testing for germline mutations in these breast carcinoma susceptibility genes. [See editorial on pages 2251-4, this issue.] Cancer 1998;83:2335-2345. © 1998 American Cancer Society.

The neural crest is of fundamental importance in the development of the head and peripheral nervous system, in the evolution of the vertebrates, and clinically because it gives rise to developmental abnormalities and neoplasms in humans, We have established a resource for studying the development of the neural crest by systematically constructing cDNA libraries horn spatiotemporally exact neural crest and related cell populations, Neural crest populations were obtained from vagal and thoracic axial levels and from branchial arches, at premigratory and early and late migratory stages, at localization stages, and after differentiation into dorsal root ganglion cells, Schwann cells, sympathetic neurons, adrenal medullary cells, and melanocytes. Libraries were constructed using several methods developed to approach the issues of making representative libraries from small amounts of tissue, The fidelity and usefulness Elf the libraries were tested, and this revealed that they expressed a variety of sequences such as integrins, CAMs, growth factors and their receptors, protein-tyrosine kinases, and phosphatases. Differential display also revealed a unique combination of cDNA species; We then selected libraries spatio-temporally appropriate for epithelium-mesenchyme transformation and probed for TGF-beta-related sequences, As anticipated, we confirmed the presence of TGF-beta 2 and dorsalin-1 but could not detect TGF-beta 1. We also revealed new expression sites, defined by the origin of the libraries, of receptors known to be expressed elsewhere (Tsk 7L; TBRII), We anticipate that this collection of cDNA libraries will be of use in studying normal and abnormal neural crest development, by both homology searches and differential expression approaches, with spatiotemporal expression information being inherent in the initial screen. (C) 1996 Academic Press, Inc.

Aims: To examine AGR2 expression in ovarian epithelial tumours and its potential role as a prognostic biomarker. Methods: Tissue microarray technology and immunohistochemistry were used to investigate AGR2 expression in ovarian epithelial tumours and in non-neoplastic ovarian epithelium. For the carcinomas, the expression data were correlated with clinicopathological features and disease outcome. Results: AGR2 was expressed in all benign, borderline and malignant mucinous tumours and in a high proportion of endometrioid carcinomas (89%). AGR2 was frequently expressed in benign and borderline serous tumours (76% and 95%, respectively), but less commonly expressed in serous carcinomas (19%, p

Summary Aims Placental mesenchymal dysplasia (PMD) is a rare condition which is associated with the disparate fetal outcomes of Beckwith-Wiedemann syndrome (BWS), fetal growth restriction or intrauterine and neonatal death. We aimed to investigate the potential epigenetic/genetic anomalies associated with PMD and their relationship with the different causes of BWS. Methods Eight archival cases in which PMD, BWS or both were diagnosed were investigated by correlating morphology with p57 KIP2 expression, XY fluorescence in situ hybridisation (FISH) analysis and DNA genotyping. Results Placentae from BWS cases caused by aberrant IC2 methylation, leading to abnormal p57 KIP2 expression, did not show PMD but had a striking excess of extravillous trophoblast. PMD in the absence of BWS was caused by androgenetic/biparental mosaicism. The single case of BWS with PMD was due to mosaic uniparental disomy of 11p15.5. In the latter two aetiologies, our results indicate that the uniparental disomy is confined to the villous mesenchyme. Conclusions These results suggest that the link between PMD and BWS is uniparental disomy of genes confined to the telomeric IC1 region of 11p15.5. A strong candidate gene is IGF2, a known growth factor of placental mesenchyme.

pression, whereas the other contained two clonal populations and was controlled only after treatment with antineoplastic chemotherapy. Conclusion. This study has demonstrated two cases of PTLPD that were derived from donor lymphoid tissue. Although both cases were associated with EBV and remained localized to allograft tissue, their clonality and response to therapy differed. Cancer 1994;74:2436-41.

HER2 is the target of a new treatment for metastatic breast cancer using the humanised monoclonal antibody trastuzumab (Herceptin). Since only around 20% of breast cancers carry the overexpressed HER2 receptor protein to which this treatment is directed, patient selection is very important in determining eligibility for the drug. Currently, immunohistochemistry and fluorescence in situ hybridisation are the main tests used for HER2 detection, and these testing recommendations have been developed based on national and international data.

To compare the morphologic and immunohistochemical properties of breast carcinomas from Chinese and Australian women in order to define possible biologic differences between these carcinomas.Three hundred cases of breast carcinomas were assessed for histologic and immunophenotypic characteristics from the pathology archives of the Changhai and St. Vincent's Hospitals.The Chinese women had proportionally more grade 2 and 3 tumors, whereas Australian women had a higher proportion of grade 1 tumor. There was a higher proportion of younger patients with a larger tumor and patients with lymph node involvement in the Chinese group as compared with Australian women. There was no difference in rate of estrogen receptor positive tumors between the 2 groups. p53 Expression was statistically more common with less cyclin D1 expression in Chinese as compared with Australian women.This study indicates that both inherent tumor biology and stage at presentation influence adversely affect the outcome of breast carcinoma in Chinese compared as with Australian women.

The endocytosis of the T cell differentiation antigen CD4 has been investigated in CD4-transfected HeLa cells, the promyelocytic HL-60 cell line, and in a number of leukemia- or lymphoma-derived T cell lines. CD4 internalization was followed using radioiodinated antibodies in an acid-elution endocytosis assay, or by covalently modifying cell surface proteins with biotin and analyzing CD4 distributions by immunoprecipitation; both approaches gave equivalent results. The assays demonstrated that in transfected HeLa cells and in HL-60 cells CD4 was constitutively internalized and recycled in the absence of ligand. Immunogold labeling and electron microscopy demonstrated that CD4 enters cells through coated pits. In contrast to the nonlymphocytic cells, T cell lines showed very little endocytosis of CD4. Measurements of fluid phase endocytosis and morphometric analysis of the endosome compartment indicated that the endocytic capacities of HeLa and lymphoid cells are equivalent and suggested that the low level of CD4 uptake in lymphocytic cells is due to exclusion of CD4 from coated pits. This conclusion was supported by experiments using truncated CD4 molecules, lacking the bulk of the cytoplasmic domain, which were internalized equally efficiently in both transfected lymphocytes and HeLa cells. Together, these results indicate that the cytoplasmic domain of CD4 mediates the different interactions with the endocytic apparatus in lymphoid and nonlymphoid cells. We suggest that the CD4-associated lymphocyte-specific protein tyrosine kinase p56lck may be involved in preventing CD4 endocytosis in T cells.

We thank P. van der Spek, J. Hoeijmakers, S. Venitt, M. Stratton, S. Easteal, R. Sinden and T. Kunkel for discussions; L. Trute for technical assistance; M. Gardner for genetic counseling of patients; and E. Edkins for heteroduplex analysis. This work was funded by the National Health and Medical Research Council of Australia and by the Victorian Health Promotion Foundation.

Deregulated activation of STAT3 is frequently associated with many human hematological and epithelial malignancies, including gastric cancer. While exaggerated STAT3 signaling facilitates an antiapoptotic, proangiogenic, and proproliferative environment for neoplastic cells, the molecular mechanisms leading to STAT3 hyperactivation remain poorly understood. Using the gp130(Y757F/Y757F) mouse model of gastric cancer, which carries a mutated gp130 cytokine receptor signaling subunit that cannot bind the negative regulator of cytokine signaling SOCS3 and is characterized by hyperactivation of the signaling molecules STAT1 and STAT3, we have provided genetic evidence that IL-11 promotes chronic gastric inflammation and associated tumorigenesis. Expression of IL-11 was increased in gastric tumors in gp130(Y757F/Y757F) mice, when compared with unaffected gastric tissue in wild-type mice, while gp130(Y757F/Y757F) mice lacking the IL-11 ligand-binding receptor subunit (IL-11Ralpha) showed normal gastric STAT3 activation and IL-11 expression and failed to develop gastric tumors. Furthermore, reducing STAT3 activity in gp130(Y757F/Y757F) mice, either genetically or by therapeutic administration of STAT3 antisense oligonucleotides, normalized gastric IL-11 expression and alleviated gastric tumor burden. Surprisingly, the genetic reduction of STAT1 expression also reduced gastric tumorigenesis in gp130(Y757F/Y757F) mice and coincided with reduced gastric inflammation and IL-11 expression. Collectively, our data have identified IL-11 as a crucial cytokine promoting chronic gastric inflammation and associated tumorigenesis mediated by excessive activation of STAT3 and STAT1.

To assess the combined role of G-CSF, GM-CSF, and M-CSF in myeloid cell production, mice deficient in all three myeloid CSFs were generated (G−/−GM−/−M−/− mice). G−/−GM−/−M−/− mice share characteristics found in mice lacking individual cytokines: they are toothless and osteopetrotic and furthermore acquire alveolar proteinosis that is more severe than that found in either GM−/− or G−/−GM−/− mice. G−/−GM−/−M−/− mice have a significantly reduced lifespan, which is prolonged by antibiotic administration, suggesting compromised ability to control bacterial infection. G−/−GM−/−M−/− mice have circulating neutrophils and monocytes, albeit at significantly reduced numbers compared with wild-type mice, but surprisingly, have more circulating monocytes than M−/− mice and more circulating neutrophils than G−/−GM−/− mice. Due to severe osteopetrosis, G−/−GM−/−M−/− mice show diminished numbers of myeloid cells, myeloid progenitors, and B lymphocytes in the bone marrow, but have significantly enhanced compensatory splenic hemopoiesis. Although G−/−GM−/−M−/− mice have a profound deficiency of myeloid cells in the resting peritoneal cavity, the animals mount a moderate cellular response in a model of sterile peritonitis. These data establish that in the absence of G-CSF, GM-CSF, and M-CSF, additional growth factor(s) can stimulate myelopoiesis and acute inflammatory responses.

summary Considerable heterogeneity of morphology and disease outcome exists within breast cancers (BC), which likely reflects variable molecular pathogeneses within this broad clinical group. Aim To evaluate the underlying genomic alterations associated with familial, early-onset BC (EOBC) phenotypes, in order to improve the management of this disease. Methods Using hierarchical clustering of morphological and immunophenotypical parameters, 116 EOBC were stratified into six groups. Conventional and array-based comparative genomic hybridisation was used to analyse the genomic alterations. Results Specific areas of genomic imbalance were associated with individual phenotypes. The largest phenotypical group was high grade, oestrogen receptor and HER-2 negative. This group contained the majority of BRCA1 germline mutation-associated tumours and commonly showed loss of chromosomal regions 5cent-5q13, 5q14–22 and 4q28-32. High mitotic rate, an important indicator of tumour cell proliferation and poor prognosis, was associated with gain of 19p, mapped within 7 Mb of the telomere. This region contains the candidate oncogene CDC34, the protein product of which is involved in ubiquitin-mediated degradation of the cyclin-dependent kinase inhibitor, p27 Kip1 . Conclusion Phenotype-based analysis can be used to determine the genetic changes important in subtypes of BC. Further, the different morphological phenotypes could act as a cost-effective surrogate for genotypical stratification to facilitate optimal management of this disease.

Summary Aims The objective of this study was to evaluate the accuracy, ease of use and reproducibility of chromogenic in situ hybridisation (CISH) for HER2 testing by studying its inter-laboratory concordance in five Australian pathology laboratories. Methods The HER2 status of 49 breast cancers was determined by CISH twice in two different laboratories. Each sample had previously been tested by immunohistochemistry (IHC; 2+ and 3+ cases selected) and fluorescence in situ hybridisation (FISH). Participating laboratories were blinded to these test results. Oestrogen receptor (ER) status was also evaluated for each cancer. Results High correlation was observed between FISH and CISH results. No cases showing high gene amplification by FISH were scored as non-amplified by CISH (kappa coefficient=1)High correlation was observed between IHC and CISH, all IHC 3+ samples showing amplification by CISH. Inter-laboratory CISH concordance was also good (kappa coefficient=0.67). Fifty-six per cent of HER2-amplified samples tested ER positive, while 42% of ER-positive cases showed HER2 gene amplification, confirming that HER2 testing should not be confined to ER-negative breast cancers. Conclusions These findings demonstrate that CISH is a robust test to assess HER2 status in breast cancer and therefore is an important addition to the HER2 testing algorithm.

We present case of renal tubular dysgenesis, a rare fetal disorder of abnormal renal tubule development which causes anhydramnios and neonatal death. Kidneys are of normal size, with non-specific, subtle, ultrasound abnormalities, but show absence or incomplete differentiation of proximal tubules due to abnormalities of the renin angiotensin system. We highlight the need for post-mortem for accurate diagnosis, targeted genetic testing, genetic counselling and subsequent antenatal screening for this usually autosomal recessive condition.

Dysfunction of proteins involved in the G1 to S transition of the cell cycle, such as p16(INK4A) and RB1, is common in many cancer types. A screen of p16 protein expression was performed in benign, borderline, and invasive ovarian tumors, together with endometrial cancers, aligned on a tissue microarray. We observed frequent p16 overexpression in serous papillary carcinomas of ovarian and endometrial origin. An extended cohort of ovarian serous papillary carcinomas was examined to further evaluate the frequency of p16 overexpression. Strong, uniform staining in the majority of cancer cells occurred commonly in invasive serous papillary ovarian cancers, particularly in grade 3 carcinomas. RB1 protein expression abnormalities were rare. Our data indicate that abnormalities in the retinoblastoma pathway, as determined by p16 overexpression, are common in serous papillary carcinomas and are probably an early event.

Ductal carcinoma in situ represents about 20% of all tumours diagnosed within mammographic screening programs. The natural history of DCIS is poorly understood, as it cannot be observed directly. Estimates of the proportion of DCIS that progress to invasive cancer, as well as factors that may influence progression, are important for clinical management. Here we review various sources of evidence regarding the natural history of DCIS. We identified relevant publications of studies on: follow-up studies of DCIS initially misdiagnosed as benign, studies of recurrence of DCIS as invasive cancer, autopsy studies, studies of risk factors for DCIS, animal studies and studies that used mathematical models to study growth of DCIS and invasive cancer. Data sources included the MEDLINE data base, searches of articles cited in key reviews and editorials. The most direct evidence regarding the progression of DCIS to invasive cancer comes from studies where DCIS was initially misdiagnosed as benign and treated by biopsy alone. These studies suggest that between 14–53% of DCIS may progress to invasive cancer over a period of 10 or more years. The reported prevalence of undiagnosed DCIS in autopsy studies, of approximately 9%, has been used to suggest a larger reservoir of DCIS may exist in the population. All types of study designs reviewed had limitations that may bias the estimate of progression in either direction. The available evidence suggests not all DCIS will progress to invasive cancer in the medium term but precise estimates of progression are not possible given the limitations of the data. Mathematical modelling of various scenarios of progression and studies of genetic factors involved in progression may shed further light on the natural history of DCIS.

Current stocks of the LCC15-MB cell line, which we originally isolated from a human breast-bone metastasis, were found to be genetically matched to the MDA-MB-435 cell line from the Lombardi Cancer Center (MDA-MB-435-LCC) using comparative genomic hybridisation, DNA microsatellite analysis and chromosomal number. LCC15-MB stocks used for our previously published studies as well as the earliest available LCC15-MB cells also showed identity to MDA-MB-435-LCC cells. The original karyotype reported for LCC15-MB cells was considerably different to that of MDA-MB-435 cells, indicating that the original LCC15-MB cells were lost to contamination by MDA-MB-435-LCC cells. Chromosome number is the simplest test to distinguish original LCC15-MB cells (n approximately 75) from MDA-MB-435 (n approximately 52). Collectively, our results prove that LCC15-MB cells currently available are MDA-MB-435 cells and we suggest their re-designation as MDA-MB-435-LCC15 cells. We also review the known misclassification of breast and prostate cancer cell lines to date and have initiated a register maintained at http://www.svi.edu.au/cell_lines_registry.doc.

Tissue microarrays are a recent innovation in the field of pathology. They were originally designed as a high-throughput approach for researchers to assess the expression of interesting candidate disease-related genes or gene products simultaneously on hundreds of tissue samples. However, their use is becoming more widespread in routine pathology, for example for quality assurance and for the optimisation of diagnostic reagents such as monoclonal antibodies and gene probes. Several molecular and conventional pathological techniques can be performed on a single tissue array, thereby enabling morphology, DNA, RNA and protein targets to be analysed on sequential sections through multiple tissue samples. Moreover, compared with full-face tissue sections, tissue microarrays are a cost- and time-efficient, effective approach to analysing biomarker expression on a large number of samples. Whilst tissue microarrays are available from commercial sources, many pathology laboratories prefer to make in-house arrays from their often extensive pathology archive to facilitate the correlation of their findings with clinical parameters. The technical skills necessary to produce tissue arrays are well within the capacity of most laboratories. However, several pitfalls to successful array production exist. The present article describes the applications of this technique and details practical points for optimal tissue array production.

Loss of genetic material from chromosome arm 8p occurs frequently in human breast carcinomas, consistent with this region of the genome harboring one or more tumor suppressor genes (TSGs). We used the complementary techniques of microsatellite-based LOH, high-density FISH, and conventional CGH on 6 breast cancer cell lines (MCF7, SKBR3, T47D, MDA MB453, BT549, and BT474) to investigate the molecular cytogenetic changes occurring on chromosome 8 during tumorigenesis, with particular emphasis on 6 potential TSGs on 8p. We identified multiple alterations of chromosome 8, including partial or complete deletion of 8p or 8q, duplication of 8q, and isochromosome 8q. The detailed FISH analysis showed several complex rearrangements of 8p with differing breakpoints of varying proximity to the genes of interest. High rates of LOH were observed at markers adjacent to or within PCM1, DUSP4/MKP2, NKX3A, and DLC1, supporting their status as candidate TSGs. Due to the complex ploidy status of these cell lines, relative loss of 8p material detected by CGH did not always correlate with microsatellite-based LOH results. These results extend our understanding of the mechanisms accompanying the dysregulation of candidate tumor suppressor loci on chromosome arm 8p, and identify appropriate cellular systems for further investigation of their biological properties.

We have identified that StarD10, a member of the START protein family, is overexpressed in both mouse and human breast tumors. StarD10 was initially discovered on the basis of its cross-reactivity with a phosphoserine-specific antibody in mammary tumors from Neu/ErbB2 transgenic mice and subsequently isolated from SKBR3 human breast carcinoma cells using a multistep biochemical purification strategy. We have shown that StarD10 is capable of binding lipids. StarD10 was found to be overexpressed in 35% of primary breast carcinomas and 64% of human breast cancer cell lines, correlating with their ErbB2/Her2 status. Coexpression of StarD10 with ErbB1/epidermal growth factor receptor in murine fibroblasts enhanced anchorage-independent growth in soft agar, providing evidence for functional cooperation between StarD10 and ErbB receptor signaling. Taken together, these data suggest that overexpression of this lipid-binding protein contributes to breast oncogenesis.

Chromosome region 17q12-23 commonly shows an increase in DNA copy number in breast cancers, suggesting that several oncogenes are located at this site. We performed a high-resolution expression array and comparative genomic hybridization analysis of genes mapped to the entire 17q12-23 region, to identify novel candidate oncogenes. We identified 24 genes that showed significant overexpression in breast cancers with gain of 17q12-23, compared to cancers without gain. These genes included previously identified oncogenes, together with several novel candidate oncogenes. FISH analysis using specific gene probes hybridized to tissue arrays confirmed the underlying amplification of overexpressed genes. This high-resolution analysis of the 17q12-23 region indicates that several established and novel candidate oncogenes, including a Wnt-signaling pathway member, are amplified and overexpressed within individual primary breast cancer samples. We were also able to confirm the presence of two apparently separate and reciprocally amplified groups of genes within this region. Investigation of these genes and their functional interactions will facilitate our understanding of breast oncogenesis and optimal management of this disease.

Aim To assess the impact of use of Electronic Request Forms (ERFs) on the amount of clinical information accompanying placentas for histopathology in a tertiary public hospital. Background In 2009 Mater Pathology Services introduced a ‘tick box' dedicated placental request form (DPRF) which significantly increased the amount of clinical information provided with placentas sent for histopathology. In 2011 ERFs were introduced across the campus, replacing the DPRF. Methods Histopathology request forms from sequential accessions of placentas were evaluated for number of pieces of clinically relevant information, defined as data that would change specimen handling or information that was used for clinicopathological correlation. ERF were compared to two control groups from different years: consecutive DPRF from 2009 and consecutive accessions using standard blank pathology request forms from 2006. Each request form was assessed by at least two authors. Results Use of the ERF significantly decreased the number of items of useful clinical information provided, compared to the DPRF (mean of 2.6 vs 1.3 items, p = 0.0004). Provision of relevant clinical information declined to the level seen prior to the introduction of the DPRF. Discussion ERFs may improve quality by reducing error, but also change clinician behaviour, resulting in provision of fewer items of useful clinical information to guide specimen handling, pathology reporting and clinicopathological correlation.

A twin pregnancy is described in which an umbilical cord hemangioma, polyhydramnios, developmental abnormalities and perinatal death were restricted to one twin, while the other twin was unaffected. Cord hemangiomas are rare and their association with fetal abnormalities is controversial. This case study supports a direct association between the cord hemangioma and the adverse pregnancy outcome, since congenital abnormalities and a cord hemangioma were present in only one of the twins.

Differences in gene expression are likely to explain the phenotypic variation between hormone-responsive and hormone-unresponsive breast cancers. In this study, DNA microarray analysis of approximately 10,000 known genes and 25,000 expressed sequence tag clusters was performed to identify genes induced by estrogen and repressed by the pure antiestrogen ICI 182 780 in vitro that correlated with estrogen receptor (ER) expression in primary breast carcinomas in vivo. Stanniocalcin (STC) 2 was identified as one of the genes that fulfilled these criteria. DNA microarray hybridization showed a 3-fold induction of STC2 mRNA expression in MCF-7 cells inor = 3 h of estrogen exposure and a 3-fold repression in the presence of antiestrogen (one-way ANOVA, P0.0005). In 13 ER-positive and 12 ER-negative breast carcinomas, the microarray-derived mRNA levels observed for STC2 correlated with tumor ER mRNA (Pearson's correlation, r = 0.85; P0.0001) and ER protein status (Spearman's rank correlation, r = 0.73; P0.0001). The expression profile of STC2 was further confirmed by in situ hybridization and immunohistochemistry on a larger cohort of 236 unselected breast carcinomas using tissue microarrays. STC2 mRNA and protein expression were found to be associated with tumor ER status (Fisher's exact test, P0.005). The related gene, STC1, was also examined and shown to be associated with ER status in breast carcinomas (Fisher's exact test, P0.05). This study demonstrates the feasibility of using global gene expression data derived from an in vitro model to pinpoint novel estrogen-responsive genes of potential clinical relevance.

LMO4 belongs to a family of LIM-only transcriptional regulators, the first two members of which are oncoproteins in acute T cell leukemia. We have explored a role for LMO4, initially described as a human breast tumor autoantigen, in developing mammary epithelium and breast oncogenesis. Lmo4 was expressed predominantly in the lobuloalveoli of the mammary gland during pregnancy. Consistent with a role in proliferation, forced expression of this gene inhibited differentiation of mammary epithelial cells. Overexpression of LMO4 mRNA was observed in 5 of 10 human breast cancer cell lines. Moreover, in situ hybridization analysis of 177 primary invasive breast carcinomas revealed overexpression of LMO4 in 56% of specimens. Immunohistochemistry confirmed overexpression in a high percentage (62%) of tumors. These studies imply a role for LMO4 in maintaining proliferation of mammary epithelium and suggest that deregulation of this gene may contribute to breast tumorigenesis.

To investigate the role of the Lyn kinase in establishing signaling thresholds in hematopoietic cells, a gain-of-function mutation analogous to the Src Y527F-activating mutation was introduced into the Lyn gene. Intriguingly, although Lyn is widely expressed within the hematopoietic system, these mice displayed no propensity toward hematological malignancy. By contrast, analysis of aging cohorts of both loss- and gain-of-function Lyn mutant mice revealed that Lyn−/− mice develop splenomegaly, increased numbers of myeloid progenitors, and monocyte/macrophage (Mφ) tumors. Biochemical analysis of cells from these mutants revealed that Lyn is essential in establishing ITIM-dependent inhibitory signaling and for activation of specific protein tyrosine phosphatases within myeloid cells. Loss of such inhibitory signaling may predispose mice lacking this putative protooncogene to tumorigenesis.

The Muir-Torre syndrome (MTS) is an autosomal dominantly inherited disorder, characterized by visceral malignancies and sebaceous skin lesions. In a subset of MTS families the disease is due to an underlying DNA mismatch-repair defect. We have identified a MTS family whose spectrum of reported neoplasia included adenocarcinomas of numerous gastrointestinal sites, carcinomas of the endometrium, ovary and breast, papillary transitional cell carcinoma of the ureter, a range of cutaneous tumors, as well as keratoacanthomas. All tumors were tested for microsatellite instability and immunohistochemically stained for expression of MLH1 and MSH2 proteins. All tumors were found to be microsatellite unstable and lacking in MSH2 protein expression. The subsequent mutation detection focused on hMSH2, and a germline mutation was identified (CAA-->TAA, Gln-->STOP, codon 337). This mutation was subsequently found in a family member with a single skin lesion only. We propose that the combination of immunohistologic and microsatellite instability analysis can be exploited to screen individuals with characteristic skin lesions even before development of visceral tumors and to direct the subsequent germline mutation search. The profile of microsatellite instability and the genes rendered dysfunctional differed between tumor samples, suggesting that the molecular pathogenesis varied between lesions, despite a common germline mutation.

Genetic changes orchestrated by human papillomaviruses are the most important known factors in carcinogenesis of the uterine cervix. However, it is clear that additional genetic events are necessary for tumour progression. We have used comparative genomic hybridization to document non-random chromosomal gains and losses within a subset of 37 cervical carcinomas matched for clinical stage Ib, but with different lymph node status. There were significantly more chromosomal changes in the primary tumours when the lymph nodes were positive for metastases. The most frequent copy number alterations were loss of 3p, 11q, 6q and 10q and gain of 3q. The smallest areas of loss and gain on chromosome 3 were 3p14–22 and 3q24–26. The study identifies progressive DNA copy number changes associated with early-stage invasive cervical cancers with and without lymph node metastases, a factor of potential prognostic and therapeutic value. © 2000 Cancer Research Campaign http://www.bjcancer.com

Secondary palate formation is a complex process that is frequently disturbed in mammals, resulting in the birth defect cleft palate. Gene targeting has identified components of cytokine/growth factor signalling systems such as Tgf-alpha/Egfr, Eph receptors B2 and B3 (Ephb2 and Ephb3, respectively), Tgf-beta2, Tgf-beta3 and activin-betaA (ref. 3) as regulators of secondary palate development. Here we demonstrate that the mouse orphan receptor 'related to tyrosine kinases' (Ryk) is essential for normal development and morphogenesis of craniofacial structures including the secondary palate. Ryk belongs to a subclass of catalytically inactive, but otherwise distantly related, receptor protein tyrosine kinases (RTKs). Mice homozygous for a null allele of Ryk have a distinctive craniofacial appearance, shortened limbs and postnatal mortality due to feeding and respiratory complications associated with a complete cleft of the secondary palate. Consistent with cleft palate phenocopy in Ephb2/Ephb3-deficient mice and the role of a Drosophila melanogaster Ryk orthologue, Derailed, in the transduction of repulsive axon pathfinding cues, our biochemical data implicate Ryk in signalling mediated by Eph receptors and the cell-junction-associated Af-6 (also known as Afadin). Our findings highlight the importance of signal crosstalk between members of different RTK subfamilies.

D-type cyclins are important cell cycle regulators that promote cellular proliferation in response to growth factors by inactivation of the retinoblastoma protein (Rb). Cyclin D1 has been shown to be overexpressed in several cancer types and to act as an oncogene in breast cancers. As D-type cyclins are rate limiting for progression into S phase, the level at which they accumulate must be carefully regulated. Several mechanisms leading to overexpression of cyclin D1 have been reported including amplification, translocation and stabilization of the mRNA. Here, we present data showing elevated cyclin D1 protein in breast cancer samples in the absence of elevated mRNA level. Further, we found that in these cases, cyclin D3 protein also accumulates and that the coordinate increase in cyclin D1 and D3 occurs in 15% (7/47) of breast cancers. In addition we show that blocking the activity of the 26S proteosome results in the accumulation of cyclin D1 and D3, that both D-type cyclins are ubiquitinated and associate with Cul-1, a component of the SCF ubiquitin ligase complex. Finally, we show that the coordinated elevation of cyclin D1 and D3 is also observed in the breast cell line MCF-7 and demonstrate that the degradation of cyclin D1 and D3 is deficient in this cell line. These results indicate that cyclin D1 and cyclin D3 share a common mechanism of degradation and we propose that the coordinate increase of D-type cyclins observed in primary breast cancers reflects a defect in their proteolysis.

Breast epithelial inclusions are the most frequently encountered ectopic structures within axillary lymph nodes. Various non‐neoplastic proliferative changes have been reported in these lesions. We report the first case of ductal carcinoma in situ (DCIS) in an axillary lymph node inclusion.

Thirty-five pediatric lymphomas were categorized as either Burkitt's lymphoma (BL), lymphoblastic lymphoma (LL), or large cell anaplastic lymphoma (LCAL) by histological and immunophenotypic methods. They were further characterized by molecular analysis of their antigen receptor genes. Southern blot (SB) and polymerase chain reaction (PCR) techniques were compared in the detection of immunogloblin heavy chain gene (IgH) rearrangement. T cell receptor beta (TCR beta) rearrangements were analyzed by SB and TCR gamma gene rearrangements by PCR. The PCR method of IgH and TCR gamma gene analysis was preferred to the SB methods, because there were fewer equivocal results in IgH gene analysis, TCR gamma rearrangement was more frequently detected than TCR beta in both lymphoblastic and large cell anaplastic lymphomas, and the PCR technique was more rapid, required less DNA, and could be used with archival material. In addition, analysis of IgH gene rearrangement by PCR was more specific for assessing B cell lineage. Although most of the molecular data were easily interpreted, occasional ambiguous results were seen due to genetic events other than antigen receptor gene rearrangement affecting the genetic analysis. Thus, it is imperative to interpret these genetic data in the context of adequate morphological and immunophenotypic analysis.

Ewing's sarcoma (ES) and other primitive peripheral neuroectodermal tumors (pPNETs) can present a significant diagnostic problem, as they may morphologically resemble other small round cell tumors (SRCTs) of childhood. However, ES/pPNET is known to carry a characteristic t(11;22)(q24;q12), the detection of which may aid diagnosis. The recent identification of the EWS and FLI-1 genes flanking the translocation break point has enabled reverse transcriptase-polymerase chain reaction (RT-PCR) to be used to detect the putative chimeric transcription factor mRNA produced by the fusion gene. We have assessed the RT-PCR method of detection by examining 40 cases of ES for the presence of EWS/FLI-1 transcripts. Twenty-six (76%) of the 34 cases with intact mRNA yielded fusion transcripts. Four different transcript sizes were detected and two tumors contained two transcripts of different size. No transcripts were detected in a control group of non-ES/pPNET SRCTs. Eight cases with intact mRNA were transcript negative. The MIC2 cell surface antigen, which is reported to be present in over 95% of ES/pPNETs, was present in 32 of 33 tumors (97%), including all 24 EWS/FLI-1 transcript-positive cases examined. Hence MIC2 is a useful screen for ES, with RT-PCR detection of t(11;22) being the optimal method for confirming the diagnosis.

Three cases of placental site trophoblast tumor (PSTT) are added to 74 cases in the English language literature. One case presented with an anorexic syndrome, the other two with amenorrhea and abnormal vaginal bleeding. The three patients, whose lesions were confined to the uterus, were treated by total hysterectomy and their follow-up has been uneventful. Review of the literature showed that 62 patients with PSTT were alive and 15 had died, two as a complication of treatment. The diagnosis of PSTT may be difficult on a dilatation and currettage specimen. Metastases are a powerful indicator of adverse outcome. Prognosis, as assessed on pathologic criteria, is unreliable; however, a mitotic count of more than five mitoses per 10 high power fields is significant. Surgery is the mainstay of treatment. Hysterectomy is generally indicated, but young patients who wish to remain fertile may be treated by conservative surgery. In a few cases of progressive disease, chemotherapy has achieved remission, but generally chemotherapy and radiotherapy are ineffective. Long-term follow-up is essential as PSTT may progress after years of remission. Serum human chorionic gonadotrophin (hCG) levels are the best available marker of disease, but the disease may still progress even if hCG levels are not raised.

We report a case of a 21-year-old woman with hematopoietic, immunological, and congenital dysmorphic abnormalities, who died following rapidly progressive, disseminated Epstein-Barr virus (EBV)-associated lymphoproliferative disease (LPD). Polymerase chain reaction (PCR) amplification of formalin-fixed paraffin-embedded tissue showed differences in the clonality of each separate lymphoproliferative lesion examined, as determined by immunoglobulin heavy chain (IgH) gene rearrangement. PCR analysis also demonstrated that all lesions contained EBV genome. Since DNA had been extracted from paraffin blocks, a direct comparison of morphology and clonality could be made in each individual lesion. The evidence from this study indicates that the monoclonal tumors arose de novo in multiple sites and that the polyclonal background observed in some lesions reflected a substantial concomitant inflammatory response.

Maternal intercurrent illness is an uncommon cause of perinatal death. We present a case of thrombotic thrombocytopenic purpura (TTP), with characteristic placental abnormalities, leading to fetal death, and review the literature concerning the onset of the rare condition during pregnancy. A 25-year-old G1P0 woman presented at 20 weeks gestation with a one day history of headaches, dark urine and decreased fetal movements. Initial assessment showed fetal heart activity present. Concurrent investigation revealed thrombocytopenia, and haemo-lytic anaemia consistent with TTP. A fetal morphology scan was performed several days after admission which revealed fetal death. Placental examination revealed large areas of acute infarction with widespread maternal vascular thrombosis. Some maternal vessels showed the characteristic finding of alternating zones of vascular dilatation and constriction (the ‘snowman sign’) consistent with aneurysmal dilatation of maternal spiral arteries with segmental intramural hyaline deposits. The placental findings directly link the TTP to the fetal death. TTP is characterised by absence or inactivation of the plasma protease (ADAMTS13) which breaks down von Willebrand factor multimers. Deficiency leads to platelet aggregation and thrombus formation. In the placenta these changes are seen in conjunction with the effects of trophoblast on the spiral arterioles producing the characteristic histological appearance. TTP rarely occurs during pregnancy, but is associated with a high rate of perinatal death.

Aims Confined placental mosaicism (CPM) is a risk factor for intrauterine growth restriction (IUGR). We investigated if we could identify pregnancies affected by high level tetraploidy (>22% of metaphases) as a form of CPM (TCPM) by clinical or placental histological features, within a larger group of pregnancies affected by IUGR of unknown origin. Methods Placentae from pregnancies affected by IUGR of unknown cause over a period of 28 months were karyotyped: 12 of 52 placentas showed TCPM. Maternal age, gravidity, parity, pre-pregnancy body mass index (BMI), and placental histology were compared between IUGR affected pregnancies with or without TCPM. Placentas with other karyotypic abnormalities were excluded from analysis. Results Parity, gravidity or maternal age showed no significant difference. IUGR without TCPM mothers tended to have a slightly but significantly higher BMI. Placental histology failed to show a significant difference with regard to the presence of abnormality or the type of parenchymal abnormality identified. Conclusion The presence of TCPM in IUGR cannot be predicted by either clinical or histological factors. Karyotyping of all placentas from pregnancies with apparently idiopathic IUGR may be advisable to identify this cause of IUGR.

Aim To investigate the expression of the novel anti-apoptotic markers GRP78 and MUC13 in extramammary Paget’s disease (EMPD). Method Twelve archival cases of EMPD, accessioned from 2001 until 2010 were assessed for expression of bcl-2, GRP78 and MUC13 using standard immunohistochemical and immunofluor-escent techniques. Results Twelve cases of EMPD were examined, four of which included an invasive component. Eight of twelve cases (67%) were positive for bcl2. Eight of eleven cases (72%), and four of eight cases (50%) with available tissue were positive for GRP78 and MUC13, respectively. Discussion Bcl2 is a well known regulator of apoptosis. GRP78 overexpression has been shown to significantly increase resistance to apoptosis in vitro and in vivo. MUC13 is overexpressed in a range of epithelial malignancies, and has poorly defined anti-apoptotic properties, as well as a role in proliferation activity via the p38MAPK signalling pathway, and anti-adhesive roles promoting cell motility. Our study has demonstrated an overexpression of the anti-apoptotic markers GRP78, MUC13, and bcl2 in EMPD. Overexpression of these anti-apoptotic markers is a novel finding in this disease and likely to be important in the pathogenesis of EMPD.

Aims To assess the impact of a dedicated placental pathology request form upon the provision of clinical information by clinicians in a tertiary hospital. Methods Consecutive series of placental pathology reports from before (98 cases) and after (92 cases) introduction of the request form were analysed for the amount of clinical information provided. Information was assessed as being clinically relevant or irrelevant. Results Use of the new form significantly improved recording of gestational age by clinicians (67% prior and 93% post). Clinicians using the new form gave significantly more information (1.57 points prior and 2.83 points post) and more of this information was considered useful in specimen handling and clinicopathological correlation (1.2 points prior and 2.1 points post). Discussion Relevant clinical information is vital to correct specimen handling and clinicopathological correlation. Implementation of a dedicated placenta request form significantly increases the frequency of recording of gestational age and the amount of information providedbyclinicians. Importantly, use of the form increases the amount of relevant information offered by referring clinicians. Increased relevant information assists in the ordering of appropriate ancillary tests, reduces time spent chasing clinical information and adds to the overall relevance and quality of placental pathology reports.

A malignant strumal carcinoid with widespread metastases resembling a well-differentiated thyroid follicular carcinoma is described. Strumal carcinoid is a rare ovarian tumor which usually behaves in a benign manner. Only one malignant case has been reported in the literature and in that case the carcinoid element metastasized. The clinical, histological, and immunohistochemical findings of the current case together with the relationship between strumal carcinoids and struma ovarii are discussed.

To our knowledge, this is the fifth reported case of invasive cervical neoplasia associated with idiopathic inflammatory myopathy and illustrates the importance of cervical examination in women with polymyositis.

Background Secretory breast carcinoma is an extremely rare subtype of invasive ductal carcinoma, accounting for less than 0.15% of all breast cancers. It has characteristic morphologic features and is the only epithelial breast tumour with a specific translocation (ETV6-NTRK3). Despite being a low grade carcinoma, it has recently been found to be express basal markers and hence is part of the basal-like spectrum of breast cancer. Case presentation We present a case of secretory carcinoma in the left breast of a 52-year-old woman. The immunophenotype was ‘triple negative’ (i.e., ER, PR and HER2 negative) with expression of some basal markers (CK5/6, vimentin and S100). Conclusion This is a rare tumour and is the only case of secretory breast carcinoma diagnosed at our institution within the last 10 years. It is important to recognise that not all carcinomas with triple negative, basal cytokeratin positive phenotype have high grade morphology.

The Royal College of Pathologists of Australasia Quality Assurance Programs (RCPA QAP) Breast Module supplies cases of all types of breast pathology to participants. In 2008 there were 280 participants, the majority from Australia ( Each year, 30 cases of breast pathology, both benign and malignant, are selected from routine diagnostic practice. Cases are initially selected by the convenor, which are supplied to a panel of expert breast pathologists for assessment. Final selection then takes place by the convenor. In 2009 the format moved entirely to virtual images. Virtual imaging is essential if breast cores and small lesions are to be included in the program. In 2008 there were 7047 responses out of a possible 8200 (86%), similar to the response rate of 2007 (88%). On average, for the 30 cases, 83% of responses were concordant and 5% assessed as minor discordant. Discordant responses accounted for approximately 10%.

The lymphoid differentiation antigen CD4 is a glycoprotein which binds to non-polymorphic regions on MHC class II molecules and is expressed primarily on helper/inducer T-lymphocytes and on some cells of the macrophage/monocyte lineage. CD4 is involved in T-cell ontogeny in the thymus and in the activation of peripheral blood lymphocytes (Robey and Axel, 1990). The specific association of CD4 with the cytoplasmic protein tyrosine kinase p56 lck (Turner et al., 1990) suggests that CD4 may be directly involved in signal transduction. In addition CD4 is used as a receptor by the human immunodeficiency viruses HIV-1 and 2. In order to gain a better understanding of the mechanisms underlying the normal function of CD4 in T-cells and in the pathway of HIV entry, we have studied the endocytic trafficking of CD4 in non-lymphoid cells transfected with the CD4 gene and in a number of leukaemia/lymphoma-derived T-cell lines.