27 results on '"Lisa Happerfield"'
Search Results
2. Supplementary Table H from Improved grading and survival prediction of human astrocytic brain tumors by artificial neural network analysis of gene expression microarray data
- Author
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V. Peter Collins, Panayiota Poirazi, Tom C. Freeman, Lisa Happerfield, Karen Plant, Lu Liu, Matthew T. Wayland, Magnus Backlund, Anastasis Oulas, and Lawrence P. Petalidis
- Abstract
Supplementary Table G from Improved grading and survival prediction of human astrocytic brain tumors by artificial neural network analysis of gene expression microarray data
- Published
- 2023
3. Data from Improved grading and survival prediction of human astrocytic brain tumors by artificial neural network analysis of gene expression microarray data
- Author
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V. Peter Collins, Panayiota Poirazi, Tom C. Freeman, Lisa Happerfield, Karen Plant, Lu Liu, Matthew T. Wayland, Magnus Backlund, Anastasis Oulas, and Lawrence P. Petalidis
- Abstract
Histopathologic grading of astrocytic tumors based on current WHO criteria offers a valuable but simplified representation of oncologic reality and is often insufficient to predict clinical outcome. In this study, we report a new astrocytic tumor microarray gene expression data set (n = 65). We have used a simple artificial neural network algorithm to address grading of human astrocytic tumors, derive specific transcriptional signatures from histopathologic subtypes of astrocytic tumors, and asses whether these molecular signatures define survival prognostic subclasses. Fifty-nine classifier genes were identified and found to fall within three distinct functional classes, that is, angiogenesis, cell differentiation, and lower-grade astrocytic tumor discrimination. These gene classes were found to characterize three molecular tumor subtypes denoted ANGIO, INTER, and LOWER. Grading of samples using these subtypes agreed with prior histopathologic grading for both our data set (96.15%) and an independent data set. Six tumors were particularly challenging to diagnose histopathologically. We present an artificial neural network grading for these samples and offer an evidence-based interpretation of grading results using clinical metadata to substantiate findings. The prognostic value of the three identified tumor subtypes was found to outperform histopathologic grading as well as tumor subtypes reported in other studies, indicating a high survival prognostic potential for the 59 gene classifiers. Finally, 11 gene classifiers that differentiate between primary and secondary glioblastomas were also identified. [Mol Cancer Ther 2008;7(5):1013–24]
- Published
- 2023
4. Abnormal placental CD8
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Susanne, Lager, Ulla, Sovio, Elizabeth, Eddershaw, Margaretha W, van der Linden, Cansu, Yazar, Emma, Cook, Lisa, Happerfield, Flora A, Jessop, Neil J, Sebire, D Stephen, Charnock-Jones, and Gordon C S, Smith
- Subjects
Fetal Growth Retardation ,Pre-Eclampsia ,Pregnancy ,Placenta ,Infant, Newborn ,Endothelial Cells ,Humans ,Female ,CD8-Positive T-Lymphocytes - Abstract
Placental pathological abnormalities are more frequently observed in complicated pregnancies than in healthy pregnancies. Infiltration of CD8Fetal growth restriction (FGR) and pre-eclampsia are severe, adverse pregnancy outcomes. Alterations in placental histology are frequently reported in these pregnancy complications and are often based upon scoring by pathologists. However, many alterations are also observed in placenta from uncomplicated pregnancies. Moreover, knowledge of disease state may bias assessment. We sought to perform an objective comparison of placental microscopic appearance in normal and complicated pregnancies. Placental villous tissue (n = 823) and edge biopsies (n = 488) from 871 individual, singleton pregnancies were collected after delivery. Cases of small-for-gestational age (SGA) or pre-eclampsia were matched with healthy controls. A subset of the SGA cases displayed signs of FGR. Cases of preterm delivery were also included. Tissue sections were stained with haematoxylin and eosin or antibodies for CD8, CD14, CD31, CD79α and elastase. Images were scored by two experienced pathologists for pathological features or analysed by image analysis and stereology. Analyses were performed blind to case-control status and gestational age. Volume fraction of T-cells increased in placentas from pregnancies complicated by pre-eclampsia (adjusted odds ratio (aOR) 1.46, 95% CI: 1.12-1.90) and FGR (aOR 1.64, 95% CI: 1.11-2.43), whereas B-cells only increased in FGR (aOR 1.65, 95% CI: 1.05-2.60). Pathological abnormalities in villous tissue were reported in 21.4% (88/411) of complicated pregnancies and 14.3% (52/363) of controls (OR 1.62, 95% CI: 1.12-2.37). There were no differences in the fractions of endothelial cells, fibrin deposition, macrophages and neutrophils when comparing normal and complicated pregnancies. In conclusion, FGR and pre-eclampsia are associated with T-cell infiltration of the placenta and placental pathological abnormalities.
- Published
- 2020
5. Abnormal placental CD8+ T cell infiltration is a feature of fetal growth restriction and pre-eclampsia
- Author
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Lisa Happerfield, F. A. Jessop, Ulla Sovio, Cansu Yazar, Elizabeth Eddershaw, Neil J. Sebire, Susanne Lager, Gordon C. S. Smith, D. Stephen Charnock-Jones, Emma Cook, Margaretha W van der Linden, Lager, Susanne [0000-0003-3556-065X], Sovio, Ulla [0000-0002-0799-1105], Yazar, Cansu [0000-0003-2660-0111], Cook, Emma [0000-0002-6142-4443], Sebire, Neil J [0000-0001-5348-9063], Charnock-Jones, D Stephen [0000-0002-2936-4890], Smith, Gordon CS [0000-0003-2124-0997], and Apollo - University of Cambridge Repository
- Subjects
0301 basic medicine ,CD31 ,Pathology ,medicine.medical_specialty ,Physiology ,leukocytes ,Placenta ,Reproduktionsmedicin och gynekologi ,CD8-Positive T-Lymphocytes ,03 medical and health sciences ,small for gestational age ,villitis of unknown aetiology ,0302 clinical medicine ,Pre-Eclampsia ,Pregnancy ,image analysis ,Obstetrics, Gynecology and Reproductive Medicine ,medicine ,Humans ,small-for-gestational age ,fibrin ,Pathological ,villous tissue ,Fetal Growth Retardation ,Eclampsia ,business.industry ,Infant, Newborn ,Endothelial Cells ,Gestational age ,medicine.disease ,endothelial cells ,030104 developmental biology ,medicine.anatomical_structure ,stereology ,Small for gestational age ,Female ,pregnancy complication ,business ,pathologist scoring ,Infiltration (medical) ,030217 neurology & neurosurgery - Abstract
KEY POINTS: Placental pathological abnormalities are more frequently observed in complicated pregnancies than in healthy pregnancies Infiltration of CD8+ T-cells into the placental villous tissue occurred in both fetal growth restriction and pre-eclampsia, whereas CD79α+ B-cell infiltration was only apparent with reduced fetal growth. Vascularisation, fibrin depositions, macrophage and neutrophil infiltration in the placenta did not differ between healthy and complicated pregnancies. ABSTRACT: Fetal growth restriction (FGR) and pre-eclampsia are severe, adverse pregnancy outcomes. Alterations in placental histology are frequently reported in these pregnancy complications and are often based upon scoring by pathologists. However, many alterations are also observed in placenta from uncomplicated pregnancies. Moreover, knowledge of disease state may bias assessment. We sought to perform an objective comparison of placental microscopic appearance in normal and complicated pregnancies. Placental villous tissue (n = 823) and edge biopsies (n = 488) from 871 individual, singleton pregnancies were collected after delivery. Cases of small-for-gestational-age (SGA) or pre-eclampsia were matched with healthy controls. A subset of the SGA cases displayed signs of FGR. Cases of pre-term delivery were also included. Tissue sections were stained with H&E or antibodies for CD8, CD14, CD31, CD79α, elastase. Images were scored by two experienced pathologists for pathological features or analysed by image analysis and stereology. Analyses were performed blind to case-control status and gestational age. Volume fraction of T-cells increased in placentas from pregnancies complicated by pre-eclampsia (adjusted odds ratio (aOR) 1.46, 95% CI 1.12-1.90) and FGR (aOR 1.64, 95% CI 1.11-2.43), whereas B-cells only increased in FGR (aOR 1.65, 95% CI 1.05-2.60). Pathological abnormalities in villous tissue were reported in 21.4% (88/411) of complicated pregnancies and 14.3% (52/363) of controls (OR 1.62, 95% CI 1.12-2.37). There were no differences in the fractions of endothelial cells, fibrin deposition, macrophages, and neutrophils when comparing normal and complicated pregnancies. In conclusion, FGR and pre-eclampsia are associated with T-cell infiltration of the placenta and placental pathological abnormalities. This article is protected by copyright. All rights reserved.
- Published
- 2020
6. Translating
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Ruth T, Casey, Mary A, McLean, Basetti, Madhu, Benjamin G, Challis, Rogier, Ten Hoopen, Thomas, Roberts, Graeme R, Clark, Deborah, Pittfield, Helen L, Simpson, Venkata R, Bulusu, Kieran, Allinson, Lisa, Happerfield, Soo-Mi, Park, Alison, Marker, Olivier, Giger, Eamonn R, Maher, and Ferdia A, Gallagher
- Subjects
Article - Abstract
PURPOSE: Mutations in the mitochondrial enzyme succinate dehydrogenase (SDH) subunit genes are associated with a wide spectrum of tumours including phaeochromocytoma and paraganglioma (PPGL) 1, 2, gastrointestinal stromal tumours (GIST) 3, renal cell carcinoma (RCC) 4 and pituitary adenomas5. SDH-related tumorigenesis is believed to be secondary to accumulation of the oncometabolite succinate. Our aim was to investigate the potential clinical applications of MRI spectroscopy ((1)H-MRS) in a range of suspected SDH-related tumours. PATIENTS AND METHODS: Fifteen patients were recruited to this study. Respiratory-gated single-voxel (1)H-MRS was performed at 3T to quantify the content of succinate at 2.4 ppm and choline at 3.22 ppm. RESULTS: A succinate peak was seen in six patients, all of whom had a germline SDHx mutation or loss of SDHB by immunohistochemistry. A succinate peak was also detected in two patients with a metastatic wild-type GIST (wtGIST) and no detectable germline SDHx mutation but a somatic epimutation in SDHC. Three patients without a tumour succinate peak retained SDHB expression, consistent with SDH functionality. In six cases with a borderline or absent peak, technical difficulties such as motion artefact rendered (1)H-MRS difficult to interpret. Sequential imaging in a patient with a metastatic abdominal paraganglioma demonstrated loss of the succinate peak after four cycles of [(177)Lu]-DOTATATE, with a corresponding biochemical response in normetanephrine. CONCLUSIONS: This study has demonstrated the translation into clinical practice of in vivo metabolomic analysis using (1)H-MRS in patients with SDH-deficient tumours. Potential applications include non-invasive diagnosis and disease stratification, as well as monitoring of tumour response to targeted treatments.
- Published
- 2019
7. Familial Adrenocortical Carcinoma in Association With Lynch Syndrome
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Benjamin G, Challis, Narayanan, Kandasamy, Andrew S, Powlson, Olympia, Koulouri, Anand Kumar, Annamalai, Lisa, Happerfield, Alison J, Marker, Mark J, Arends, Serena, Nik-Zainal, and Mark, Gurnell
- Subjects
congenital, hereditary, and neonatal diseases and abnormalities ,MutS Homolog 2 Protein ,Neoplastic Syndromes, Hereditary ,Adrenocortical Carcinoma ,Humans ,Female ,Genetic Predisposition to Disease ,Genetic Testing ,Middle Aged ,Special Features ,Colorectal Neoplasms, Hereditary Nonpolyposis ,Adrenal Cortex Neoplasms ,Germ-Line Mutation - Abstract
Context: Adrenocortical carcinoma (ACC) is a rare endocrine malignancy with a poor prognosis. Although the majority of childhood ACC arises in the context of inherited cancer susceptibility syndromes, it remains less clear whether a hereditary tumor predisposition exists for the development of ACC in adults. Here, we report the first occurrence of familial ACC in a kindred with Lynch syndrome resulting from a pathogenic germline MSH2 mutation. Case: A 54-year-old female with a history of ovarian and colorectal malignancy was found to have an ACC. A detailed family history revealed her mother had died of ACC and her sister had previously been diagnosed with endometrial and colorectal cancers. A unifying diagnosis of Lynch syndrome was considered, and immunohistochemical analyses demonstrated loss of MSH2 and MSH6 expression in both AACs (proband and her mother) and in the endometrial carcinoma of her sister. Subsequent genetic screening confirmed the presence of a germline MSH2 mutation (resulting in deletions of exons 1–3) in the proband and her sister. Conclusion: Our findings provide strong support for the recent proposal that ACC should be considered a Lynch syndrome-associated tumor and included in the Amsterdam II clinical diagnostic criteria. We also suggest that screening for ACC should be considered in cancer surveillance strategies directed at individuals with germline mutations in DNA mismatch repair genes., This study provides the first description of an intergenerational occurrence of ACC in a family with Lynch Syndrome resulting from a germline MSH2 mutation. Thus, ACC should be included in clinical diagnostic criteria for LS and considered in cancer surveillance recommendations for individuals with germline mutations in DNA mismatch repair genes.
- Published
- 2016
8. Microarray, qPCR, andKCNJ5Sequencing of Aldosterone-Producing Adenomas Reveal Differences in Genotype and Phenotype between Zona Glomerulosa- and Zona Fasciculata-Like Tumors
- Author
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Elena A.B. Azizan, Gary J. Hoffman, Brian Y.H. Lam, Junhua Zhou, Alison Marker, Rhoda E. Kuc, Jennifer Clarke, Morris J. Brown, Lisa Happerfield, and Stephen Newhouse
- Subjects
Adenoma ,Adult ,Male ,Familial hyperaldosteronism ,medicine.medical_specialty ,Genotype ,Endocrinology, Diabetes and Metabolism ,education ,Clinical Biochemistry ,Adrenal Gland Neoplasms ,Biology ,Biochemistry ,chemistry.chemical_compound ,Endocrinology ,Zona fasciculata ,Internal medicine ,Hyperaldosteronism ,mental disorders ,medicine ,Humans ,Aldosterone ,Aged ,Laser capture microdissection ,Adrenal gland ,Biochemistry (medical) ,Middle Aged ,medicine.disease ,Molecular biology ,Phenotype ,medicine.anatomical_structure ,G Protein-Coupled Inwardly-Rectifying Potassium Channels ,chemistry ,Zona glomerulosa ,Female ,Zona Glomerulosa ,Zona Fasciculata ,psychological phenomena and processes - Abstract
Aldosterone-producing adenomas (APA) are heterogeneous. The recent finding of somatic KCNJ5 mutations suggests a genetic explanation.The objectives of this study were the following: 1) to compare transcriptional profiles in APA and adjacent adrenal gland (AAG); 2) to test whether gene expression profile clusters with different cell histology; and 3) to measure the frequency of KCNJ5 mutations and determine the genotype-phenotype relationship.The design of the study included laboratory analyses of 46 unselected APA.The patients in this study had primary hyperaldosteronism with unilateral APA.The objectives of this study were the following: 1) Illumina beadchip analysis of RNA from eight paired APA-AAG; 2) a blinded review of cell histology for 46 APA; 3) laser capture microdissection of zona glomerulosa (ZG) and zona fasciculata (ZF) cells; and 4) sequencing of KCNJ5 in 46 APA.The main outcome measures of this study were the following: 1) a difference in gene expression profile and a correlation with histological markers of ZF; 2) a frequency of KCNJ5 mutations and phenotypic comparisons of wild type with mutant APA.The results of the study were the following: 1) a cluster analysis of microarray data separated APA from AAG. APA at opposite ends of the APA cluster had an approximately 800-fold difference in CYP17A1 mRNA expression, whereas histology showed 0% ZF-like cells in one vs. 100% in the other. A heat map ranking APA by CYP17A1 expression correctly predicted several genes (e.g. KCNK1, SLC24A3) to be enriched in laser capture microdissection samples of ZG; 2) known or novel mutations of KCNJ5 were found in 20 of 46 consecutive APA [43% (95% confidence interval [CI] (29, 58)%)]. The APA with KCNJ5 gene mutations were larger compared with tumors harboring the wild type, 1.63 [95% CI (1.37, 1.88)] vs. 1.14 [0.97, 1.30] cm (P = 0.0013), had predominantly ZF-like cells, and their CYP17A1 (log(2)-fold change) was higher than in wild type: -0.96 [95% CI (-0.07, -1.85)] vs. -2.54 [-1.61, -3.46], (P = 0.017).KCNJ5 mutations are common in APA, particularly those arising from ZF. The long-recognized heterogeneity among APA may have a genetic basis.
- Published
- 2012
9. Multilocus Inherited Neoplasia Alleles Syndrome: A Case Series and Review
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Eamonn R. Maher, Derek Lim, Rick van Minkelen, Emma R. Woodward, Marc Tischkowitz, Ian M. Frayling, Lone Sunde, James Whitworth, Lisa Happerfield, Anne-Bine Skytte, Mark J. Arends, Clinical Genetics, Whitworth, James [0000-0002-3682-2298], Tischkowitz, Marc [0000-0002-7880-0628], Maher, Eamonn [0000-0002-6226-6918], and Apollo - University of Cambridge Repository
- Subjects
0301 basic medicine ,Adult ,Male ,Cancer Research ,Databases, Factual ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Genetic disorder diagnosis ,Locus heterogeneity ,Neoplasms ,Proto-Oncogene Proteins ,Genotype ,medicine ,Humans ,Genetic Predisposition to Disease ,Allele ,Exome sequencing ,Alleles ,Aged ,Genetics ,Tumor Suppressor Proteins ,Cancer ,High-Throughput Nucleotide Sequencing ,Syndrome ,Middle Aged ,medicine.disease ,Penetrance ,030104 developmental biology ,MutS Homolog 2 Protein ,Oncology ,030220 oncology & carcinogenesis ,Mutation ,Allelic heterogeneity ,Female ,Tumor Suppressor Protein p53 - Abstract
Mendelian causes of inherited cancer susceptibility are mostly rare and characterized by variable expression and incomplete penetrance. Phenotypic variability may result from a range of causes including locus heterogeneity, allelic heterogeneity, genetic and environmental modifier effects, or chance. Another potential cause is the presence of 2 or more inherited cancer predisposition alleles in the same individual. Although the frequency of such occurrences might be predicted to be low, such cases have probably been underascertained because standard clinical practice has been to test candidate inherited cancer genes sequentially until a pathogenic mutation is detected. However, recent advances in next-generation sequencing technologies now provide the opportunity to perform simultaneous parallel testing of large numbers of inherited cancer genes. Herein we provide examples of patients who harbor pathogenic mutations in multiple inherited cancer genes and review previously published examples to illustrate the complex genotype-phenotype relationships in these cases. We suggest that clinicians should proactively consider the likelihood of this phenomenon (referred to herein as multilocus inherited neoplasia alleles syndrome [MINAS]) in patients with unusual inherited cancer syndrome phenotypes. To facilitate the clinical management of novel cases of MINAS, we have established a database to collect information on what is likely to be an increasingly recognized cohort of such individuals.
- Published
- 2015
10. MLH1 Promoter Methylation, Diet, and Lifestyle Factors in Mismatch Repair Deficient Colorectal Cancer Patients From EPIC-Norfolk
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Laura J. Gay, Mark J. Arends, Panagiota N. Mitrou, Richard Bowman, Ashraf E. Ibrahim, Lisa Happerfield, Robert Luben, Alison McTaggart, Richard Y. Ball, and Sheila A. Rodwell
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Adult ,Male ,Oncology ,congenital, hereditary, and neonatal diseases and abnormalities ,Cancer Research ,medicine.medical_specialty ,Pathology ,Colorectal cancer ,Medicine (miscellaneous) ,Biology ,MLH1 ,DNA Mismatch Repair ,Surveys and Questionnaires ,Internal medicine ,Vegetables ,medicine ,Humans ,Prospective Studies ,Promoter Regions, Genetic ,Prospective cohort study ,Life Style ,neoplasms ,Adaptor Proteins, Signal Transducing ,Aged ,Nutrition and Dietetics ,Smoking ,Nuclear Proteins ,Microsatellite instability ,Promoter ,DNA ,Methylation ,DNA Methylation ,Middle Aged ,medicine.disease ,Immunohistochemistry ,United Kingdom ,digestive system diseases ,Diet ,Fruit ,DNA methylation ,Female ,Microsatellite Instability ,DNA mismatch repair ,Colorectal Neoplasms ,MutL Protein Homolog 1 - Abstract
There is conflicting evidence for the role diet and lifestyle play in the development of mismatch repair (MMR)-deficient colorectal cancers (CRC). In this study, associations between MMR deficiency, clinicopathological characteristics, and dietary and lifestyle factors in sporadic CRC were investigated. Tumor samples from 185 individuals in the EPIC-Norfolk study were analyzed for MLH1 gene promoter methylation and microsatellite instability (MSI). Dietary and lifestyle data were collected prospectively using 7-day food diaries (7dd) and questionnaires. MMR-deficient tumor cases (MLH1 promoter methylation positive, MSI-H) were more likely to be female, older at diagnosis, early Dukes' stage (A/B), and proximal in location (MSI-H P = 0.03, 0.03, 0.02, and 0.001, respectively). Tumors with positive MLH1 promoter methylation (>20%) were associated with poor differentiation (P = 0.03). Low physical activity was associated with cases without MSI (P = 0.05). MMR deficiency was not significantly associated with cigarette smoking or alcohol, folate, fruit, vegetable, or meat consumption. We conclude that MMR-deficient tumors represent a distinct subset of sporadic CRC that are proximal in location, early Dukes' stage, and poorly differentiated, in cases that are female and older at diagnosis. There is no overall role for diet and lifestyle in MMR status in CRC, consistent with age-related susceptibility to MLH1 promoter methylation.
- Published
- 2011
11. Phospho-STAT5 and phospho-Akt expression in chronic myeloproliferative neoplasms
- Author
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Lizz F. Grimwade, Colin Tristram, Lisa Happerfield, Gary McIntosh, Paul Scorer, Amy E. Quinn, Michael A. Scott, Wendy N. Erber, Nigel H. Piggott, Marie Hall, Elaine M. Boyd, Mark Rees, and A Bench
- Subjects
Male ,Bone Marrow Cells ,Biology ,Exon ,Mastocytosis, Systemic ,hemic and lymphatic diseases ,STAT5 Transcription Factor ,medicine ,Humans ,Phosphorylation ,Systemic mastocytosis ,Protein kinase B ,PI3K/AKT/mTOR pathway ,STAT5 ,Aged ,Myeloproliferative Disorders ,Kinase ,food and beverages ,Hematology ,Janus Kinase 2 ,Middle Aged ,medicine.disease ,Neoplasm Proteins ,Proto-Oncogene Proteins c-kit ,Chronic Disease ,biology.protein ,Cancer research ,Female ,Signal transduction ,Megakaryocytes ,Proto-Oncogene Proteins c-akt ,Receptors, Thrombopoietin - Abstract
The majority of Myeloproliferative Neoplasms (MPNs) are characterised by mutations in genes encoding molecules or receptors involved in cell signalling, the most common being the JAK2 V617F mutation. This mutation leads to ligand-independent activation of downstream signalling pathways by constitutive phosphorylation. The signalling pathways affected include the Janus kinase-signal transducers and activators of transcription (JAK-STAT) and phosphotidylinositide-3 kinase (PI3K) pathways, which regulate cell survival and apoptosis respectively. Monoclonal antibodies to phospho-STAT5 and phospho-Akt were generated and assessed by immunocytochemistry on bone marrow biopsies of MPN patients with JAK2 V617F, JAK2 exon 12, MPL exon 10 and KIT D816V mutations. JAK2 V617F mutation was associated with significantly increased levels of phosphorylated STAT5 and Akt in haemopoietic cells, most marked in megakaryocytes. In contrast, JAK2 exon 12 and MPL exon 10 mutations did not affect the level of phosphorylation. In systemic mastocytosis with KIT D618V mutation there was significantly increased expression of phosphorylated STAT5 and Akt in neoplastic mast cells although there was no change in the expression in other haemopoietic cells. JAK2 V617F is associated with upregulated phosphorylation of STAT5 and Akt in megakaryocytes, and to a lesser extent in other haemopoietic cells. Immunocytochemistry of bone marrow trephines for these phospho-proteins can be used as a supplementary diagnostic test with a high negative predictive value.
- Published
- 2009
12. Megaoesophagus inRassf1a-null mice
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David J. Adams, Lisa Happerfield, Louise van der Weyden, and Mark J. Arends
- Subjects
Pathology ,medicine.medical_specialty ,Achalasia ,Biology ,Pathology and Forensic Medicine ,Pathogenesis ,Mice ,Esophagus ,Fibrosis ,medicine ,Animals ,Genetic Predisposition to Disease ,Molecular Biology ,Myenteric plexus ,Mice, Knockout ,Tumor Suppressor Proteins ,S100 Proteins ,Histology ,Original Articles ,Cell Biology ,medicine.disease ,Esophageal Achalasia ,Disease Models, Animal ,medicine.anatomical_structure ,Immunohistochemistry ,Chronic Inflammatory Infiltrate - Abstract
Megaoesophagus, or oesophageal achalasia, is a neuromuscular disorder characterized by an absence of peristalsis and flaccid dilatation of the oesophagus, resulting in the retention of ingesta in the dilated segment. The aetiology and pathogenesis of idiopathic (or primary) megaoesophagus are still poorly understood and very little is known about the genetic causes of megaoesophagus in humans. Attempts to develop animal models of this condition have been largely unsuccessful and although the ICRC/HiCri strain of mice spontaneously develop megaoesophagus, the underlying genetic cause remains unknown. In this report, we show that aged Rassf1a-null mice have an enhanced susceptibility to megaoesophagus compared with wild-type littermates (approximately 20%vs. approximately 2% incidence respectively; P = 0.01). Histological examination of the dilated oesophaguses shows a reduction in the numbers of nerve cells (both ganglia and nerve fibres) in the myenteric plexus of the dilated mid and lower oesophagus that was confirmed by S100 immunohistochemistry. There was also a chronic inflammatory infiltrate and subsequent fibrosis of the myenteric plexus and the muscle layers. These appearances closely mimic the gross and histopathological findings in human cases of megaoesophagus/achalasia, thus demonstrating that this is a representative mouse model of the disease. Thus, we have identified a genetic cause of the development of megaoesophagus/achalasia that could be screened for in patients, and may eventually facilitate the development of therapies that could prevent further progression of the disease once it is diagnosed at an early stage.
- Published
- 2009
13. Improved grading and survival prediction of human astrocytic brain tumors by artificial neural network analysis of gene expression microarray data
- Author
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Matthew T. Wayland, Lawrence P. Petalidis, Anastasis Oulas, Lisa Happerfield, Lu Liu, Karen Plant, V. Peter Collins, Tom C. Freeman, Panayiota Poirazi, and Magnus Bäcklund
- Subjects
Cancer Research ,Angiogenesis ,Computational biology ,Astrocytoma ,Biology ,Bioinformatics ,Article ,Text mining ,Biomarkers, Tumor ,medicine ,Humans ,Grading (tumors) ,Survival rate ,Oligonucleotide Array Sequence Analysis ,Regulation of gene expression ,Brain Neoplasms ,business.industry ,Astrocytic Tumor ,Gene Expression Profiling ,Intracellular Signaling Peptides and Proteins ,Reproducibility of Results ,Neural Networks (Computer) ,Phosphoproteins ,Prognosis ,medicine.disease ,Gene Expression Regulation, Neoplastic ,Survival Rate ,Gene expression profiling ,Oncology ,Tumor Markers, Biological ,Neural Networks, Computer ,Apoptosis Regulatory Proteins ,business ,Algorithms - Abstract
Histopathologic grading of astrocytic tumors based on current WHO criteria offers a valuable but simplified representation of oncologic reality and is often insufficient to predict clinical outcome. In this study, we report a new astrocytic tumor microarray gene expression data set (n = 65). We have used a simple artificial neural network algorithm to address grading of human astrocytic tumors, derive specific transcriptional signatures from histopathologic subtypes of astrocytic tumors, and asses whether these molecular signatures define survival prognostic subclasses. Fifty-nine classifier genes were identified and found to fall within three distinct functional classes, that is, angiogenesis, cell differentiation, and lower-grade astrocytic tumor discrimination. These gene classes were found to characterize three molecular tumor subtypes denoted ANGIO, INTER, and LOWER. Grading of samples using these subtypes agreed with prior histopathologic grading for both our data set (96.15%) and an independent data set. Six tumors were particularly challenging to diagnose histopathologically. We present an artificial neural network grading for these samples and offer an evidence-based interpretation of grading results using clinical metadata to substantiate findings. The prognostic value of the three identified tumor subtypes was found to outperform histopathologic grading as well as tumor subtypes reported in other studies, indicating a high survival prognostic potential for the 59 gene classifiers. Finally, 11 gene classifiers that differentiate between primary and secondary glioblastomas were also identified. [Mol Cancer Ther 2008;7(5):1013–24]
- Published
- 2008
14. Pregnancy, Primary Aldosteronism, and Adrenal CTNNB1 Mutations
- Author
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Junhua Zhou, Mariann Bienz, Sumedha Garg, Morris J. Brown, Lalarukh Haris Shaikh, Elena A.B. Azizan, Alison Marker, Fiona E. Karet Frankl, Mark Gurnell, Ada E. D. Teo, and Lisa Happerfield
- Subjects
Adenoma ,Adult ,medicine.medical_specialty ,Beta-catenin ,Somatic cell ,Adrenal Gland Neoplasm ,Adrenal Gland Neoplasms ,Hypokalemia ,Article ,Primary aldosteronism ,Pregnancy ,Internal medicine ,Hyperaldosteronism ,medicine ,Humans ,Aldosterone ,beta Catenin ,biology ,business.industry ,GNRHR ,Wnt signaling pathway ,General Medicine ,Middle Aged ,Receptors, LH ,medicine.disease ,Up-Regulation ,Postmenopause ,Endocrinology ,Hypertension ,Cancer research ,biology.protein ,Female ,business ,Pregnancy Complications, Neoplastic ,Receptors, LHRH - Abstract
Recent discoveries of somatic mutations permit the recognition of subtypes of aldosterone-producing adenomas with distinct clinical presentations and pathological features. Here we describe three women with hyperaldosteronism, two who presented in pregnancy and one who presented after menopause. Their aldosterone-producing adenomas harbored activating mutations of CTNNB1, encoding β-catenin in the Wnt cell-differentiation pathway, and expressed LHCGR and GNRHR, encoding gonadal receptors, at levels that were more than 100 times as high as the levels in other aldosterone-producing adenomas. The mutations stimulate Wnt activation and cause adrenocortical cells to de-differentiate toward their common adrenal-gonadal precursor cell type. (Funded by grants from the National Institute for Health Research Cambridge Biomedical Research Centre and others.).
- Published
- 2015
15. Effect of somatic CTNNB1 mutations on adrenocortical cell de-differentiation in adenoma presenting in early pregnancy or menopause
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Mariann Bienz, Lisa Happerfield, Alison Marker, Elena A.B. Azizan, Mark Gurnell, Sumedha Garg, Morris J. Brown, F.E. Karet Frankl, L. Haris Shaikh, Ada E. D. Teo, and Junhua Zhou
- Subjects
0301 basic medicine ,endocrine system ,medicine.medical_specialty ,Mutation ,Microarray ,Adenoma ,Microarray analysis techniques ,Endocrinology, Diabetes and Metabolism ,Receptor expression ,Wnt signaling pathway ,General Medicine ,Biology ,medicine.disease ,medicine.disease_cause ,Transcriptome ,03 medical and health sciences ,030104 developmental biology ,Endocrinology ,Primary aldosteronism ,Internal medicine ,medicine - Abstract
Background Primary aldosteronism is a curable cause of hypertension if the hypertension is due to an aldosterone-producing adenoma (APA) [1] , [2] . The discovery of somatic mutations defining pathogenetic subtypes of this adenoma has increased recognition of common but small zona glomerulosa-like APAs [3] , [4] , [5] , [6] . In this subtype, we have sought APAs with distinctive presentation or outcome, in which analysis of genotype–phenotype associations could help explain the low success rate and variability of adrenalectomy for curing hypertension. Methods Ten zona glomerulosa-like APAs from three women with primary aldosteronism were analysed by exome sequencing and microarray analysis. Nine had a mutation of ATP1A1 or CACNA1D, and one had a distinctive genotype and transcriptome. Targeted sequencing of further zona glomerulosa-like APAs led to the finding of two with a similar genotype, and to the recognition that all three women presented in early pregnancy or menopause. Quantitative PCR and immunohistochemistry were performed for genes upregulated in the index case. Similar analyses and a TCF–LEF assay for Wnt signalling were performed on cells transfected with the mutant gene. Findings All three APAs had novel mutations in exon 3 of CTNNB1, encoding β-catenin in the Wnt cell-differentiation pathway. The mutations prevented phosphorylation of β-catenin, resulting in an increase in total form of β-catenin and active to total ratio, as quantified by immunoblotting. Transfection of mutant constructs into HEK cells caused near doubling of Wnt signalling in a luciferase reporter assay. Microarray of the index case found a greater than 100-fold increase in expression of gonadal receptors LHCGR and GNRHR, confirmed by qPCR (both genes) and immunohistochemistry (LHCGR) in all three APAs. Increased GATA4 expression was further evidence of adrenocortical cell de-differentiation towards their common gonad-adrenal precursor cell-type. Transfection of LHCGR-negative APA cells with GFP-mutant-CTNNB1 switched on LHCGR expression in GFP-positive cells. Interpretation The role of the Wnt system in adrenal physiology and tumour development is well recognised [7] , [8] . De-differentiation of zona glomerulosa-cells after Wnt activation by the CTNNB1 mutation seems to cause aberrant gonadal receptor expression, which is also previously described [9] , [10] . Our findings connect these observations, and explain the unmasking of small APAs by increases in luteinising hormone and human chorionic gonadotropin hormone in early pregnancy or menopause. Strikingly, all three women, including the treatment-resistant older woman, were clinically cured by adrenalectomy. This contrasts with the failure of adrenalectomy to completely cure hypertension in the majority of patients. This failure is attributable to decades of exposure to aldosterone excess, and the secondary consequences of fibrosis and remodelling in the cardiovascular system [11] , [12] . CTNNB1 mutations have now been reported in 10% of APAs without a previously reported mutation [13] . Elucidation of APA subtypes could, with preoperative genotyping, permit recognition of patients in whom recent onset of primary aldosteronism is likely to be completely resolved by surgery.
- Published
- 2016
16. Pathological Society of Great Britain and Ireland. 182nd meeting, 3-5 January 2001
- Author
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Mark J. Arends, M. Jimenez-Linan, Ian Martin Frayling, and Lisa Happerfield
- Subjects
Oncology ,medicine.medical_specialty ,business.industry ,Colorectal cancer ,Normal tissue ,Cancer ,MLH1 ,medicine.disease ,Pathology and Forensic Medicine ,MSH2 ,Internal medicine ,medicine ,business ,Immunostaining - Published
- 2001
17. Familial Adrenocortical Carcinoma Associated with HNPCC
- Author
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Narayanan Kandasamy, Serena Nik-Zainal, Anand Kumar Annamalai, Lisa Happerfield, Joan Paterson, Mark Arends, and Mark Gurnell
- Published
- 2011
18. Alterations in PTEN and PIK3CA in colorectal cancers in the EPIC Norfolk study: associations with clinicopathological and dietary factors
- Author
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Adam Naguib, James C Cooke, Lisa Happerfield, Lucy Kerr, Laura J Gay, Robert N Luben, Richard Y Ball, Panagiota N Mitrou, Alison McTaggart, Mark J Arends, Luben, Robert [0000-0002-5088-6343], and Apollo - University of Cambridge Repository
- Subjects
Male ,Questionnaires ,Cancer Research ,Class I Phosphatidylinositol 3-Kinases ,DNA Mutational Analysis ,Gene mutation ,Adenocarcinoma ,medicine.disease_cause ,Proto-Oncogene Mas ,lcsh:RC254-282 ,03 medical and health sciences ,Phosphatidylinositol 3-Kinases ,0302 clinical medicine ,Sex Factors ,Surgical oncology ,Surveys and Questionnaires ,medicine ,Genetics ,PTEN ,Humans ,neoplasms ,PI3K/AKT/mTOR pathway ,Genetic Association Studies ,030304 developmental biology ,Aged ,Regulation of gene expression ,Aged, 80 and over ,0303 health sciences ,Mutation ,biology ,PTEN Phosphohydrolase ,Middle Aged ,medicine.disease ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Immunohistochemistry ,Gene Expression Regulation, Neoplastic ,Oncology ,030220 oncology & carcinogenesis ,Colonic Neoplasms ,Cancer research ,biology.protein ,Disease Progression ,Female ,Research Article - Abstract
Background The PTEN tumour suppressor gene and PIK3CA proto-oncogene encode proteins which contribute to regulation and propagation of signal transduction through the PI3K/AKT signalling pathway. This study investigates the prevalence of loss of PTEN expression and mutations in both PTEN and PIK3CA in colorectal cancers (CRC) and their associations with tumour clinicopathological features, lifestyle factors and dietary consumptions. Methods 186 adenocarcinomas and 16 adenomas from the EPIC Norfolk study were tested for PTEN and PIK3CA mutations by DNA sequencing and PTEN expression changes by immunohistochemistry. Dietary and lifestyle data were collected prospectively using seven day food diaries and lifestyle questionnaires. Results Mutations in exons 7 and 8 of PTEN were observed in 2.2% of CRC and PTEN loss of expression was identified in 34.9% CRC. Negative PTEN expression was associated with lower blood low-density lipoprotein concentrations (p = 0.05). PIK3CA mutations were observed in 7% of cancers and were more frequent in CRCs in females (p = 0.04). Analysis of dietary intakes demonstrated no link between PTEN expression status and any specific dietary factor. PTEN expression negative, proximal CRC were of more advanced Dukes' stage (p = 0.02) and poor differentiation (p < 0.01). Testing of the prevalence of PIK3CA mutations and loss of PTEN expression demonstrated that these two events were independent (p = 0.55). Conclusion These data demonstrated the frequent occurrence (34.9%) of PTEN loss of expression in colorectal cancers, for which gene mutations do not appear to be the main cause. Furthermore, dietary factors are not associated with loss of PTEN expression. PTEN expression negative CRC were not homogenous, as proximal cancers were associated with a more advanced Dukes' stage and poor differentiation, whereas distal cancers were associated with earlier Dukes' stage.
- Published
- 2011
19. An immunohistochemical and fluorescence in situ hybridization-based comparison between the Oracle HER2 Bond Immunohistochemical System, Dako HercepTest, and Vysis PathVysion HER2 FISH using both commercially validated and modified ASCO/CAP and United Kingdom HER2 IHC scoring guidelines
- Author
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Lisa Happerfield, Anthony O'Grady, Lilian Tee, Nicola Johnson, Elena Provenzano, David Allen, Elaine W. Kay, Mai Gu, and Sarah E Pinder
- Subjects
Oncology ,medicine.medical_specialty ,Pathology ,Histology ,Receptor, ErbB-2 ,Concordance ,Breast Neoplasms ,Guidelines as Topic ,HercepTest ,Sensitivity and Specificity ,Oracle ,Pathology and Forensic Medicine ,Automation ,Breast cancer ,Internal medicine ,Biomarkers, Tumor ,Medicine ,Humans ,Neoplasm Invasiveness ,skin and connective tissue diseases ,In Situ Hybridization, Fluorescence ,Paraffin Embedding ,medicine.diagnostic_test ,Staining and Labeling ,business.industry ,medicine.disease ,Immunohistochemistry ,Neoplasm Proteins ,Medical Laboratory Technology ,Research Design ,%22">Fish ,Female ,business ,Asco cap ,Fluorescence in situ hybridization - Abstract
Immunohistochemistry (IHC) is used as the frontline assay to determine HER2 status in invasive breast cancer patients. The aim of the study was to compare the performance of the Leica Oracle HER2 Bond IHC System (Oracle) with the current most readily accepted Dako HercepTest (HercepTest), using both commercially validated and modified ASCO/CAP and UK HER2 IHC scoring guidelines. A total of 445 breast cancer samples from 3 international clinical HER2 referral centers were stained with the 2 test systems and scored in a blinded fashion by experienced pathologists. The overall agreement between the 2 tests in a 3×3 (negative, equivocal and positive) analysis shows a concordance of 86.7% and 86.3%, respectively when analyzed using commercially validated and modified ASCO/CAP and UK HER2 IHC scoring guidelines. There is a good concordance between the Oracle and the HercepTest. The advantages of a complete fully automated test such as the Oracle include standardization of key analytical factors and improved turn around time. The implementation of the modified ASCO/CAP and UK HER2 IHC scoring guidelines has minimal effect on either assay interpretation, showing that Oracle can be used as a methodology for accurately determining HER2 IHC status in formalin fixed, paraffin-embedded breast cancer tissue.
- Published
- 2010
20. Automated immunostaining of cell smears: an alternative to flow cytometry
- Author
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Lizz F. Grimwade, David Bloxham, Lisa Happerfield, Wendy N. Erber, and Rani Saward
- Subjects
Pathology ,medicine.medical_specialty ,Tissue Fixation ,Bone Marrow Cells ,Biology ,Pathology and Forensic Medicine ,Flow cytometry ,Immunophenotyping ,Immunoenzyme Techniques ,Antigen ,Antigens, Neoplasm ,medicine ,Biomarkers, Tumor ,Humans ,Electronic Data Processing ,Blood Cells ,Immunoperoxidase ,medicine.diagnostic_test ,General Medicine ,Alkaline Phosphatase ,Molecular biology ,Staining ,medicine.anatomical_structure ,Case-Control Studies ,Hematologic Neoplasms ,Alkaline phosphatase ,Immunohistochemistry ,Feasibility Studies ,Bone marrow ,Immunostaining - Abstract
Aims: To assess the applicability of an automated tissue immunostainer machine for the phenotypic analysis of peripheral blood and bone marrow aspirate smears. Methods: Air-dried smears of peripheral blood and bone marrow from normal individuals and 14 patients with haematological malignancies were stained, following fixation, with a range of antibodies to haemopoietic antigens using both immunoperoxidase and immuno-alkaline phosphatase methods on the Bond-maX (Leica Microsystems) automated immunostaining machine. Results: Automated protocols used for staining formalin-fixed paraffin-embedded tissue could be applied to cell smears, giving high quality staining with excellent cytological detail and antigenic preservation for an extensive antibody panel. Optimal quality (morphological preservation and antigen detection without background staining) was obtained with an alkaline phosphatase method and Fast Red chromogenic substrate. Both cell surface and intracellular (cytoplasmic and nuclear) antigens could be detected and gave the expected reactivity. Conclusions: Automated immunostaining is possible for haematological smears. It generates consistent high quality staining, with the exception of surface immunoglobulin, and is applicable to haematological and, potentially, cytological smears. It provides a practical alternative to flow cytometry and will have particular application when smears are available and there is no suitable sample for flow cytometry.
- Published
- 2008
21. Human thymocyte development in mouse organ cultures
- Author
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Deborah E. Restall, Amanda G. Fisher, Lena Larsson, Matthias Merkenschiager, Lisa Happerfield, and Lindsey K. Goff
- Subjects
Antigens, Differentiation, T-Lymphocyte ,Stromal cell ,CD8 Antigens ,T-Lymphocytes ,Immunology ,Double negative ,Thymus Gland ,Organ culture ,Mice ,Organ Culture Techniques ,Precursor cell ,Animals ,Humans ,Immunology and Allergy ,Mice, Inbred BALB C ,Chimera ,Chemistry ,Cell Differentiation ,General Medicine ,Hematopoietic Stem Cells ,Embryonic stem cell ,In vitro ,Cell biology ,Thymocyte ,Phenotype ,CD4 Antigens ,Cell Division ,CD8 - Abstract
A novel system to study human thymocyte development is described in which embryonic mouse thymic rudiments are seeded with human precursor cells in vitro. In these cultures human thymocytes proliferate extensively (greater than 20-fold increase in cell number) and mature, as evidenced by the accumulation of double and single positive (CD4+ and/or CD8+) cells. Data presented here suggest that the survival and ordered development of the mature human thymocytes in chimeric thymuses is dependent on human stromal elements. Immature CD4-CD8- human thymocytes failed to colonize or minimally recolonized mouse thymic lobes unless provided with high density (greater than 1.077 g/ml) human thymic cell fractions. These fractions contain multicellular complexes of epithelial/nurse cells, thymocytes, and dendritic cells/macrophages which dramatically enhanced the recolonizing capacity of purified CD4-CD8- thymocytes. The chimeric organ culture system described here provides not only a new approach for studying human T cell ontogeny but also a direct means for the future dissection of stromal interactions necessary for successful transition of precursor cells (CD4-CD8-) to immature double positive (CD4+CD8+) and mature single positive cells (CD4+ or CD8+) in the thymus.
- Published
- 1990
22. A novel CD11c monoclonal antibody effective in formalin-fixed tissue for the diagnosis of hairy cell leukemia
- Author
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Ioannis Anagnostopoulos, Lisa Happerfield, Harald Stein, John Brown, Wendy N. Erber, and Korinna Jöhrens
- Subjects
Pathology ,medicine.medical_specialty ,Lymphoma, B-Cell ,medicine.drug_class ,Biopsy ,CD11c ,Monoclonal antibody ,Sensitivity and Specificity ,Pathology and Forensic Medicine ,Diagnosis, Differential ,Fixatives ,Bone Marrow ,hemic and lymphatic diseases ,Formaldehyde ,medicine ,Humans ,Hairy cell leukemia ,Molecular Biology ,Leukemia, Hairy Cell ,Paraffin Embedding ,biology ,medicine.diagnostic_test ,business.industry ,fungi ,food and beverages ,Antibodies, Monoclonal ,Reproducibility of Results ,Histology ,Cell Biology ,General Medicine ,medicine.disease ,Immunohistochemistry ,Lymphoma ,CD11c Antigen ,Leukemia ,Case-Control Studies ,biology.protein ,Antibody ,business - Abstract
Objective: The diagnosis of hairy cell leukemia (HCL) and its distinction from other B-cell lymphomas can be difficult in formalin-fixed tissue. This is because the histology is not always classical, and despite having a characteristic phenotype, there are few relevant monoclonal antibodies with sufficient sensitivity and specificity that can be applied to fixed material. In this study, we assessed the utility of a newly developed antibody against a formalin-resistant CD11c epitope (5D11) for the diagnosis and monitoring of HCL in formalin-fixed tissue. Methods: We performed immunohistochemical staining for CD11c expression in formalin-fixed and also decalcified tissue of 196 small B-cell lymphomas, including 104 cases of HCL showing extensive to minimal infiltrates. Results: The CD11c antibody was both sensitive and specific for HCL, even in cases with minimal infiltration. Conclusion: We recommend that this CD11c antibody be added to a panel of antibodies for immunostaining of formalin-fixed material, for differentiation of HCL from other small B-cell lymphomas, and for detection of residual disease following therapy.
- Published
- 2007
23. Application of Molecular Diagnostics to Hereditary Nonpolyposis Colorectal Cancer
- Author
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C. Mattocks, Lisa Happerfield, Kim Oakhill, Ian M. Frayling, and Mark Arends
- Subjects
Oncology ,medicine.medical_specialty ,business.industry ,Colorectal cancer ,Endometrial cancer ,Cancer ,medicine.disease ,Molecular diagnostics ,digestive system diseases ,Lynch syndrome ,Familial adenomatous polyposis ,Cancer syndrome ,Internal medicine ,medicine ,Familial Cancer ,business - Abstract
Hereditary nonpolyposis colorectal cancer is a tumor predis– position syndrome characterised by a propensity to develop, typically, but by no means exclusively, young‐onset colorectal and other cancers (1). The condition was first described in 1913 by the US pathologist Warthin in a comprehensive survey of familial cancer (2). He was stimulated to make this study because his seamstress was depressed at the thought of dying prematurely from bowel or womb cancer, as had many of her relatives. She was a member of Family “ G” in his original arti–cle, which incidentally contains examples of most of the cancer genetic conditions recognized today (2). Family “G” was redis– covered in the 1960s by Lynch, although it was not at first real-ized that it was one of Warthins original families (3,4). Lynch later made a distinction between families with only bowel can– cer (Lynch syndrome type 1: site‐specific colorectal cancer) and families with several types of cancer, including bowel (Lynch syndrome type 2: family cancer syndrome). Given the propensity to bowel cancer, but without the polyposis charac– teristic of familial adenomatous polyposis (FAP), the all embracing term “hereditary nonpolyposis colorectal cancer” (HNPCC) is now used (1). However, having to explain all this in the clinic makes the idea of going back to calling it Lynch or family cancer syndrome rather attractive.
- Published
- 2006
24. 802 SOMATIC MUTATIONS OF KCNJ5 ARE COMMON IN ALDOSTERONE-PRODUCING ADENOMAS OF THE ADRENAL, AND DISTINGUISH A PHENOTYPE OF YOUNGER WOMEN WITH ZF-LIKE LARGE ADENOMAS FROM OLDER MEN WITH ZG-LIKE SMALL ADENOMAS
- Author
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Elena A.B. Azizan, Gary J. Hoffman, Alison Marker, Brian Y.H. Lam, Rhoda E. Kuc, Jennifer Clarke, Lisa Happerfield, Morris J. Brown, Stephen Newhouse, and Junhua Zhou
- Subjects
medicine.medical_specialty ,Aldosterone ,biology ,Physiology ,Somatic cell ,business.industry ,Phenotype ,chemistry.chemical_compound ,Endocrinology ,chemistry ,Internal medicine ,KCNJ5 ,Internal Medicine ,biology.protein ,medicine ,Cardiology and Cardiovascular Medicine ,business - Published
- 2012
25. DNA replication licensing and human cell proliferation
- Author
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L.G. Bobrow, Sergei Romanov, G. A. Thomas, Kai Stoeber, E. Dillwyn Williams, Thea D. Tlsty, Gareth H. Williams, and Lisa Happerfield
- Subjects
Origin Recognition Complex ,Fluorescent Antibody Technique ,Eukaryotic DNA replication ,Cell Cycle Proteins ,Pre-replication complex ,Medical and Health Sciences ,DNA replication factor CDT1 ,Mice ,2.1 Biological and endogenous factors ,Aetiology ,Cellular Senescence ,Cancer ,Microscopy ,Microscopy, Confocal ,Cell Cycle ,Nuclear Proteins ,Cell Differentiation ,Biological Sciences ,Cell biology ,DNA-Binding Proteins ,ORC ,Confocal ,Cell Division ,Non-programmatic ,DNA Replication ,1.1 Normal biological development and functioning ,Biology ,Cdc6 ,Cell Line ,Replication factor C ,Control of chromosome duplication ,Underpinning research ,Breast Cancer ,Genetics ,Animals ,Humans ,DNA replication licensing ,Cell-Free System ,G1 Phase ,proliferation marker ,Minichromosome Maintenance Complex Component 2 ,Cell Biology ,MCM ,Molecular biology ,MCM Protein ,Licensing factor ,cell proliferation ,Hela Cells ,biology.protein ,Origin recognition complex ,Generic health relevance ,Schizosaccharomyces pombe Proteins ,Developmental Biology ,HeLa Cells - Abstract
The convergence point of growth regulatory pathways that control cell proliferation is the initiation of genome replication, the core of which is the assembly of pre-replicative complexes resulting in chromatin being ‘licensed’ for DNA replication in the subsequent S phase. We have analysed regulation of the pre-replicative complex proteins ORC, Cdc6, and MCM in cycling and non-proliferating quiescent, differentiated and replicative senescent human cells. Moreover, a human cell-free DNA replication system has been exploited to study the replicative capacity of nuclei and cytosolic extracts prepared from these cells. These studies demonstrate that downregulation of the Cdc6 and MCM constituents of the replication initiation pathway is a common downstream mechanism for loss of proliferative capacity in human cells. Furthermore, analysis of MCM protein expression in self-renewing, stable and permanent human tissues shows that the three classes of tissue have developed very different growth control strategies with respect to replication licensing. Notably, in breast tissue we found striking differences between the proportion of mammary acinar cells that express MCM proteins and those labelled with conventional proliferation markers, raising the intriguing possibility that progenitor cells of some tissues are held in a prolonged G1 phase or ‘in-cycle arrest’. We conclude that biomarkers for replication-licensed cells detect, in addition to actively proliferating cells, cells with growth potential, a concept that has major implications for developmental and cancer biology.
- Published
- 2001
26. Evaluation of PGP9.5 in the diagnosis of Hirschsprung's disease
- Author
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Virginia R. Sams, J. W. Keeling, Lynda G. Bobrow, and Lisa Happerfield
- Subjects
Adult ,Pathology ,medicine.medical_specialty ,Muscularis mucosae ,Adolescent ,Myenteric Plexus ,Pathology and Forensic Medicine ,Random Allocation ,medicine ,Humans ,Hirschsprung Disease ,Child ,Hirschsprung's disease ,Lamina propria ,Frozen section procedure ,Megacolon ,business.industry ,Infant, Newborn ,Infant ,Reproducibility of Results ,Frozen Section Diagnosis ,medicine.disease ,Immunohistochemistry ,Staining ,Ganglion ,medicine.anatomical_structure ,Cholinergic Fibers ,Child, Preschool ,Ganglia ,Thiolester Hydrolases ,business ,Ubiquitin Thiolesterase - Abstract
The ability of an acetylcholinesterase-stained frozen section to detect an increase in large cholinergic nerve fibres within the muscularis mucosae and extending into the lamina propria was a significant step forward in the diagnosis of Hirschsprung's disease (HD). However, such frozen section diagnosis is not always possible. The purpose of this study was to assess the ability of PGP9.5 to detect this pattern of mucosal nerve fibre staining immunohistochemically. Sixty-four specimens were included in the study. Twenty-six of these had been diagnosed as HD by conventional means. All cases were stained immunohistochemically with PGP9.5, S100, and anti-neurofilaments (NF). Twenty-four cases of HD were also stained with neurone-specific enolase (NSE). PGP9.5 reliably stained fibres in the mucosal and submucosal plexuses, and ganglion cells, when the latter were present. This positive staining of ganglion cells was more intense than that seen with NSE, and the positive fibre staining was more intense than that seen with NF. Increased lamina propria fibres were detected with PGP9.5 in only 37 per cent of HD cases compared with S100 positive staining in 60 per cent of cases. However, when S100 staining was assessed alone, it gave a higher false-negative rate in diagnosing HD than PGP9.5 used alone. Therefore we would recommend the use of PGP9.5 and S100 together for the immunohistochemical diagnosis of HD in formalin-fixed biopsies.
- Published
- 1992
27. Molecular Classification of Breast Carcinomas Using Tissue Microarrays.
- Author
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Grace Callagy, Elena Cattaneo, Yataro Daigo, Lisa Happerfield, Lynda G. Bobrow, Paul D. P. Pharoah, and Carlos Caldas
- Published
- 2003
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