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ISSUE ADDRESSED: Children and adolescents are particularly receptive to cues from food and drink advertising. Several policies recommend restricting unhealthy or discretionary ('junk') food advertising on government infrastructure such as public transport. Prior research in New South Wales (NSW) and Western Australia (WA) reveals a high proportion of discretionary food advertising outdoors and on public transport. The aim of this study was to identify the landscape of food and drink advertising on public transport assets around Adelaide schools. METHODS: An environmental scan of food and drink advertising on public transport assets within 500 m of 65 Adelaide schools was conducted. The Council of Australian Governments Health Council's interim guide was used to categorise advertisements as 'suitable' or 'unsuitable' for promotion. RESULTS: Almost four in every five food and drink advertisements on public-transport assets observed around Adelaide schools included discretionary food or drink, and therefore were categorised as unsuitable for promotion. Advertisements on bus exteriors were more likely to promote discretionary foods, compared with bus shelters. CONCLUSIONS: This proportion of 'unhealthy' food advertising was comparable to previous research conducted in WA and NSW. SO WHAT?: In light of historic state/territory actions such as the ACT preventing discretionary food advertising on public transport, these findings should prompt a review and discussion of the suitability of food and drink promotion on government-owned South Australian transport and their associated assets.
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The factor VIII intron 22 inversion is the most common cause of hemophilia A, accounting for approximately 40% of all severe cases of the disease. Southern hybridization and multiplex long distance PCR are the most commonly used techniques to detect the inversion in a diagnostic setting, although both have significant limitations. Here we describe our experience establishing a multicolor fluorescent in situ hybridization (FISH) based assay as an alternative to existing methods for genetic diagnosis of the inversion. Our assay was designed to apply three differentially labelled BAC DNA probes that when hybridized to interphase nuclei would exhibit signal patterns that are consistent with the normal or the inversion locus. When the FISH assay was applied to five normal and five inversion male samples, the correct genotype was assignable with p<0.001 for all samples. When applied to carrier female samples the assay could not assign a genotype to all female samples, probably due to a lower proportion of informative nuclei in female samples caused by the added complexity of a second X chromosome. Despite this complication, these pilot findings show that the assay performs favourably compared to the commonly used methods.
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Núcleo Celular/genética , Fator VIII/genética , Hemofilia A/genética , Hibridização in Situ Fluorescente/métodos , Tipagem Molecular/métodos , Sítios de Ligação/genética , Southern Blotting , Núcleo Celular/metabolismo , Inversão Cromossômica , Cromossomos Artificiais Bacterianos/metabolismo , Cromossomos Humanos , DNA/análise , Sondas de DNA/metabolismo , Feminino , Genótipo , Hemofilia A/diagnóstico , Heterozigoto , Humanos , Íntrons , Leucócitos Mononucleares/citologia , Masculino , Reação em Cadeia da Polimerase , Fatores SexuaisRESUMO
Genetic analysis of breast tumor core needle biopsies may provide early diagnostic information that is useful to direct subsequent clinical management. We report metaphase comparative genomic hybridization (CGH) analysis of 42 core needle biopsies prospectively sampled from invasive ductal breast carcinomas of 41 patients, and show that recurrent chromosomal copy number changes are associated with histologically defined tumor features. As far as is known, this is the first time CGH profiles from diagnostic breast tumor core needle biopsies have been reported in association with pathological data in such detail. A comparison of Grade 1 (n = 7), Grade 2 (n = 16), and Grade 3 tumors (n = 19) revealed a chromosomal copy number gain involving 8q22 that was associated with higher grade (1/7 vs. 16/19, respectively) and therefore altered cell differentiation status. CGH results were validated using tumor touch imprints prepared from a subgroup from the same sample set (n = 18) by fluorescence in situ hybridization (FISH) and two probes selected from regions of interest within 8p21 and 8q22. Overall concordance between CGH and FISH for 8p21 and 8q22 imbalances was 9/18 (50%) and 12/18 (67%), respectively. When FISH and CGH data were combined, and tumors classified according to better defined resected tumor histology available for 34cases, 19/19 Grade 3 tumors showed 8q22 gain compared with 0/6 Grade 1 tumors and 4/9 Grade 2 tumors. Further comprehensive studies are required to verify the biological significance of this association and its potential to facilitate early clinicopathological assessment of breast cancer. This article contains Supplementary Material available at http://www.interscience.wiley.com/jpages/1045-2257/suppmat.
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Neoplasias da Mama/genética , Aberrações Cromossômicas , Cromossomos Humanos Par 8 , Biópsia por Agulha , Neoplasias da Mama/patologia , Estudos de Coortes , Humanos , Hibridização in Situ Fluorescente , Invasividade Neoplásica , Hibridização de Ácido NucleicoRESUMO
Purpose: One third of ER-positive breast cancer patients who initially respond to endocrine therapy become resistant to treatment. Such treatment failure is associated with poor prognosis and remains an area of unmet clinical need. Here, we identify a specific posttranslational modification that occurs during endocrine resistance and which results in tumor susceptibility to the apoptosis-inducer TRAIL. This potentially offers a novel stratified approach to targeting endocrine-resistant breast cancer.Experimental Design: Cell line and primary-derived xenograft models of endocrine resistance were investigated for susceptibility to TRAIL. Tumor viability, cancer stem cell (CSC) viability (tumorspheres), tumor growth kinetics, and metastatic burden were assessed. Western blots for the TRAIL-pathway inhibitor, c-FLIP, and upstream regulators were performed. Results were confirmed in primary culture of 26 endocrine-resistant and endocrine-naïve breast tumors.Results: Breast cancer cell lines with acquired resistance to tamoxifen (TAMR) or faslodex were more sensitive to TRAIL than their endocrine-sensitive controls. Moreover, TRAIL eliminated CSC-like activity in TAMR cells, resulting in prolonged remission of xenografts in vivo In primary culture, TRAIL significantly depleted CSCs in 85% endocrine-resistant, compared with 8% endocrine-naïve, tumors, whereas systemic administration of TRAIL in endocrine-resistant patient-derived xenografts reduced tumor growth, CSC-like activity, and metastases. Acquired TRAIL sensitivity correlated with a reduction in intracellular levels of c-FLIP, and an increase in Jnk-mediated phosphorylation of E3-ligase, ITCH, which degrades c-FLIP.Conclusions: These results identify a novel mechanism of acquired vulnerability to an extrinsic cell death stimulus, in endocrine-resistant breast cancers, which has both therapeutic and prognostic potential. Clin Cancer Res; 24(10); 2452-63. ©2018 AACR.
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Neoplasias da Mama/metabolismo , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/metabolismo , Resistencia a Medicamentos Antineoplásicos , Processamento de Proteína Pós-Traducional , Receptores de Estrogênio/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Animais , Antineoplásicos Hormonais/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/genética , Linhagem Celular Tumoral , Modelos Animais de Doenças , Resistencia a Medicamentos Antineoplásicos/genética , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Camundongos , Células-Tronco Neoplásicas/efeitos dos fármacos , Células-Tronco Neoplásicas/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Genomic rearrangements are a well-recognized cause of genetic disease and can be formed by a variety of mechanisms. We report a complex rearrangement causing severe hemophilia A, identified and further characterized using a range of PCR-based methods, and confirmed using array-comparative genomic hybridization (array-CGH). This rearrangement consists of a 15.5-kb deletion/16-bp insertion located 0.6 kb from a 28.1-kb deletion/263-kb insertion at Xq28 and is one of the most complex rearrangements described at a DNA sequence level. We propose that the rearrangement was generated by distinct but linked cellular responses to double strand breakage, namely break-induced replication (BIR) and a novel model of break-induced serial replication slippage (SRS). The copy number of several genes is affected by this rearrangement, with deletion of part of the Factor VIII gene (F8, causing hemophilia A) and the FUNDC2 gene, and duplication of the TMEM185A, HSFX1, MAGEA9, and MAGEA11 genes. As the patient exhibits no clinically detectable phenotype other than hemophilia A, it appears that the biological effects of the other genes involved are not dosage-dependent. This investigation has provided novel insights into processes of DNA repair including BIR and the first description of SRS during repair in a pathological context.
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Cromossomos Humanos X/genética , Fator VIII/genética , Mutação , Adolescente , Antígenos de Neoplasias/genética , Sequência de Bases , Análise Mutacional de DNA , Proteínas de Ligação a DNA/genética , Duplicação Gênica , Fatores de Transcrição de Choque Térmico , Proteínas de Choque Térmico/genética , Hemofilia A/genética , Hemofilia A/patologia , Humanos , Masculino , Modelos Genéticos , Mutagênese Insercional , Proteínas de Neoplasias/genética , Proteínas do Tecido Nervoso/genética , Receptores Acoplados a Proteínas G/genética , Deleção de SequênciaRESUMO
PURPOSE: Previous studies have shown that increased trans-scleral permeability after exposure to certain prostaglandins is associated with increased intrascleral matrix metalloproteinases (MMPs). The present study was undertaken to determine whether these MMPs could directly alter transscleral permeability. METHODS: Freshly enucleated mouse eyes were incubated with human MMP-1, -2, and -14 for 4 hours at 37 degrees C. The eyes then were incubated with 10 or 70 kDa dextran-tetramethylrhodamine-lysine for 16 to 32 minutes at 37 degrees C. Two methods of analysis were used. In the first, quickly isolated retinas were homogenized and centrifuged. Fluorescence in the supernatants was determined by microspectrofluorimetry. In the second, the eyes were fixed in 4% paraformaldehyde, and frozen sections were prepared. After the identity of the sections was masked, the intensity of fluorescence in anterior, middle, and posterior regions of the outer retina and inner retina was scored with a 7-point grading scheme. RESULTS: The concentration of 10-kDa fluorescent dextran was 5.14 +/- 1.61 microg/mL (mean +/- SD, n = 33) in the control retinal supernatants, and 6.37 +/- 2.67 microg/mL (n = 40) in the retinal supernatants from the MMP-treated eyes. This increase was statistically significant (P < 0.02, t-test). The structural organization of the retina and other ocular tissues was maintained in all experimental conditions. Histologic scoring of fluorescence found significantly increased dextran in the outer retina of eyes treated with MMPs for 32 minutes (the score of control eyes was 2.5 +/- 0.4 and of MMP-treated eyes was 3.5 +/- 0.1, mean +/- SD; P = 0.02, n = 3). Analysis by region found greater scores in the third of the retina nearest to the optic nerve head. CONCLUSIONS: These results show that MMP-1, -2, and -14 can directly increase transscleral permeability and support the view that the increased MMP-1 and -2 observed after topical PG treatment could contribute to increased uveoscleral outflow.
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Biotina/análogos & derivados , Dextranos/metabolismo , Metaloproteinases da Matriz/farmacologia , Retina/metabolismo , Rodaminas/metabolismo , Esclera/metabolismo , Animais , Biotina/metabolismo , Camundongos , Permeabilidade/efeitos dos fármacos , Espectrometria de FluorescênciaRESUMO
Invasive ductal carcinomas of the breast (IDC) are routinely assessed on hematoxylin and eosin stained paraffin sections, with limited use of immunohistochemistry (IHC). Most IDC are regarded as a single diagnostic entity, IDC of no special type (IDC-NST), which is subdivided further only by grading. However, recent research suggests that there is high clinical relevance in differentiating IDC subtypes. Here, we ascertain whether tumor histology alone can predict basal or luminal cell phenotype in high-grade IDC-NST, and whether IHC and molecular characteristics are associated with the observed morphologies. A total of 29 grade 3 IDC-NST samples were studied, 10 tumors from a selected pilot cohort A and 19 tumors from an unselected validation cohort B. Along with histopathological assessment, the expression of ESR1, PGR, ERBB2 (HER-2), the basal/myoepithelial marker TP73L (p63), cytokeratins 5/6 (KRT5/6) and 14 (KRT14), and the luminal-specific cytokeratins 8/18 (KRT 8/18) and 19 (KRT19) was assessed by IHC. Hierarchical cluster analysis of clinicopathological variables and, separately, microarray expression profiles showed that the phenotypically distinctive basaloid and luminal tumors of cohort A fell into two main groups, defined by heterogeneous or uniformly positive expression of KRT8/18. The 38 genes differentially expressed between these two classes included ERBB2, KRT8, and six other genes previously associated with ERBB2-positive or luminal phenotypes. Tumor histology was not predictive for validation cohort B, but quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed two molecularly defined clusters that again aligned with the KRT8/18 staining phenotypes. Metaphase comparative genomic hybridization revealed 10q, 16q, and 20q copy-number imbalances that associated recurrently with KRT8/18 staining patterns.
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Neoplasias da Mama/genética , Queratina-18/genética , Queratina-8/genética , Invasividade Neoplásica/genética , Adulto , Idoso , Estudos de Coortes , DNA Complementar , Feminino , Humanos , Pessoa de Meia-Idade , Hibridização de Ácido Nucleico , Análise de Sequência com Séries de Oligonucleotídeos , Receptores de Estrogênio/análise , Receptores de Progesterona/análise , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Changing demographic, social, economic and technological trends have impacted the expectations of the Academic Health Center in preparing physicians to serve the needs of the American society, resulting in revisions to current curricula. In addition to the traditional basic sciences and clinical disciplines, accredited medical schools are required to provide curriculum exposure in behavioral health, communication skills, diversity and cultural awareness, ethics, evidence-based medicine, geriatrics, integrative medicine, pain management, palliative care, public health, socio-economic dynamics, and domestic violence. These themes are considered 'cross-cutting' since it is recognized these important curricular components apply across all years of medical school. In this article, the authors describe a strategic model developed at the University of New Mexico School of Medicine (UNMSOM) to integrate horizontally and vertically 12 cross-cutting themes as an evolving interdisciplinary curriculum reform process. These areas were defined through a combination of internal self-study, external requirements, and student and faculty interest. In the early stage of use of this model at UNMSOM, the authors describe the new cross-cutting themes that have been integrated. Minimal disruption and a spirit of cooperation and acceptance have characterized the curricular change that has been required. Preliminary assessment indicates that the program has been successful.
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Currículo/normas , Prestação Integrada de Cuidados de Saúde , Educação de Graduação em Medicina/tendências , Modelos Educacionais , Faculdades de Medicina/normas , Centros Médicos Acadêmicos , Comportamento Cooperativo , Currículo/tendências , Educação de Graduação em Medicina/organização & administração , Avaliação Educacional , Humanos , Comunicação Interdisciplinar , New Mexico , Desenvolvimento de Programas , Avaliação de Programas e Projetos de Saúde , Inquéritos e QuestionáriosRESUMO
The activation of genes important to acute lymphoblastic leukemia (ALL) may be evidenced by somatically acquired chromosomal translocations found recurrently in different patient subgroups. It is for this reason that research efforts have focused on the molecular dissection of recurring chromosomal rearrangements. However, even though a large number of leukemia-causing genes have been identified, the genetic basis of many ALL cases remains unknown. We and others have reasoned that novel translocations found in the leukemic cells of ALL patients may mark the location of more frequent gene rearrangements that are otherwise hidden submicroscopically within normal or complex karyotypes. Towards this end, we here describe the first reported association of a t(5;10)(q22;q24) with adult ALL. Fluorescence in situ hybridization (FISH) and Southern blot hybridization studies have eliminated likely involvement of the candidate genes APC and MCC on chromosome 5, and PAX2, TLX1, and NFKB2 on chromosome 10. Results further suggest that the breakpoint on chromosome 5 lies centromeric of APC and the chromosome 10 breakpoint is centromeric of PAX2. The genomic regions disrupted by this t(5;10)(q22;q24) have not previously been associated with leukemia.
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Cromossomos Humanos Par 5 , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Translocação Genética , Células da Medula Óssea/patologia , Bandeamento Cromossômico , Mapeamento Cromossômico , Cromossomos Humanos Par 10 , Humanos , Masculino , Metáfase , Pessoa de Meia-Idade , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologiaRESUMO
OBJECTIVES: Despite advances in perioperative management during surgical treatment of hypoplastic left heart syndrome (HLHS), patients are still at risk for adverse neurodevelopmental sequelae including cognitive dysfunction. This study aimed to assess a neuropsychological profile of HLHS patients at school age who underwent the Norwood operation between 1996 and 2003 with deep hypothermic circulatory arrest (DHCA) or antegrade selective cerebral perfusion (ASCP), respectively. METHODS: Forty HLHS patients (DHCA: n = 11 and ASCP: n = 29) were compared with healthy controls (DHCA controls: n = 10 and ASCP controls: n = 24), recruited according to age, sex and socioeconomic status. Neuropsychological assessment included non-verbal intelligence (IQ norms) and raw score measures of visual and verbal short- and long-term memory as well as executive functions, processing speed and concentration. Neuropsychological data were correlated with bypass and circulatory arrest times. RESULTS: Compared with control subjects, both patient groups had average non-verbal intelligence scores [DHCA: 102 (72-112) and ASCP: 92 (70-127)], but showed reduced long-term memory capacities and decreased executive performance as well as reduced processing speed. DHCA patients, furthermore, had a reduced visual attention span, and ASCP patients scored less on the verbal learning task than controls. The duration of DHCA and cardiopulmonary bypass (CPB) was associated with visual executive functions and short-term memory. In the DHCA group, the duration of DHCA was correlated with the time to complete the Trail Making Test A measuring visual executive functions (Spearman rank correlation, rS = 0.867, P = 0.018). In patients provided with ASCP, the cumulated CPB duration was negatively correlated with the score of the block span test measuring visual short memory (rS = -0.476, P = 0.020) as well as with the copy score of the Rey Figure assessing visual executive functions (rS = -0.399, P = 0.032). CONCLUSIONS: School-age children with HLHS who underwent the Norwood procedure either with DHCA or ASCP show cognitive impairments compared with healthy controls. Our data indicate deficits in specific cognitive domains such as memory, executive functions and processing speed rather than basic intellectual dysfunction.
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Parada Circulatória Induzida por Hipotermia Profunda/métodos , Transtornos Cognitivos/etiologia , Cognição/fisiologia , Síndrome do Coração Esquerdo Hipoplásico/cirurgia , Procedimentos de Norwood/efeitos adversos , Cuidados Paliativos/métodos , Criança , Feminino , Humanos , Masculino , Memória de Curto Prazo/fisiologia , Testes Neuropsicológicos , Estudos RetrospectivosRESUMO
BACKGROUND: Chromosome band 10q24 is a gene-rich domain and host to a number of cancer, developmental, and neurological genes. Recurring translocations, deletions and mutations involving this chromosome band have been observed in different human cancers and other disease conditions, but the precise identification of breakpoint sites, and detailed characterization of the genetic basis and mechanisms which underlie many of these rearrangements has yet to be resolved. Towards this end it is vital to establish a definitive genetic map of this region, which to date has shown considerable volatility through time in published works of scientific journals, within different builds of the same international genomic database, and across the differently constructed databases. RESULTS: Using a combination of chromosome and interphase fluorescent in situ hybridization (FISH), BAC end-sequencing and genomic database analysis we present a physical map showing that the order and chromosomal orientation of selected genes within 10q24 is CEN-CYP2C9-PAX2-HOX11-NFKB2-TEL. Our analysis has resolved the orientation of an otherwise dynamically evolving assembly of larger contigs upstream of this region, and in so doing verifies the order and orientation of a further 9 cancer-related genes and GOT1. This study further shows that the previously reported human papillomavirus type 6a DNA integration site HPV6AI1 does not map to 10q24, but that it maps at the interface of chromosome bands 14q13.3-q21.1. CONCLUSIONS: This revised map will allow more precise localization of chromosome rearrangements involving chromosome band 10q24, and will serve as a useful baseline to better understand the molecular aetiology of chromosomal instability in this region. In particular, the relocation of HPV6AI1 is important to report because this HPV6a integration site, originally isolated from a tonsillar carcinoma, was shown to be rearranged in other HPV6a-related malignancies, including 2 of 25 genital condylomas, and 2 of 7 head and neck tumors tested. Our finding shifts the focus of this genomic interest from 10q24 to the chromosome 14 site.
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Cromossomos Humanos Par 10/genética , Cromossomos Humanos Par 14/genética , Proteínas de Ligação a DNA/genética , Genes Neoplásicos/genética , Proteínas de Homeodomínio/genética , NF-kappa B/genética , Proteínas Oncogênicas/genética , Papillomaviridae/genética , Mapeamento Físico do Cromossomo/métodos , Fatores de Transcrição/genética , Integração Viral/genética , Cromossomos Artificiais Bacterianos/genética , DNA Viral/genética , Ordem dos Genes/genética , Marcadores Genéticos/genética , Humanos , Hibridização in Situ Fluorescente/métodos , Subunidade p52 de NF-kappa B , Fator de Transcrição PAX2 , Infecções por Papillomavirus/genética , Proteínas Proto-Oncogênicas/genética , Análise de Sequência de DNA/métodos , Infecções Tumorais por Vírus/genéticaRESUMO
We report the case of a 61-year-old woman who presented with a myelodysplastic syndrome (MDS) and a t(5;7)(q11.2;p15) in her bone marrow cells. Subsequent analysis of phytohemagglutinin-stimulated peripheral blood lymphocytes and cultured skin fibroblasts showed that the translocation was constitutional. Disruption of chromosome bands 5q11.2 and 7p15 has been described recurrently in MDS and acute myelocytic leukemia (AML) and, although the age of onset was not earlier than usual, it is nonetheless possible that genes interrupted by this translocation may been a predisposing factor for her condition. With progression to AML, a further rearrangement of the constitutional der(7)t(5;7) occurred, involving chromosome arm 1q. Fluorescence in situ hybridization (FISH) with whole-chromosome paints showed that the result of the second rearrangement, a t(1;7)(q32.1;q32), was observed, leading to trisomy of the segment 1q32.1 approximately qter and monosomy of the segment 7q32.1 approximately qter. The acquired imbalances, particularly loss of 7q, are commonly associated with MDS/AML and a poor prognosis; however, this patient remained in remission after treatment for more than two years before AML relapse, perhaps because the affected regions fall outside of the critical regions of imbalance.
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Cromossomos Humanos Par 5 , Cromossomos Humanos Par 7 , Leucemia Mieloide Aguda/genética , Síndromes Mielodisplásicas/genética , Translocação Genética , Células da Medula Óssea , Transformação Celular Neoplásica/genética , Deleção Cromossômica , Coloração Cromossômica , Cromossomos Humanos Par 1 , Feminino , Humanos , Pessoa de Meia-Idade , TrissomiaRESUMO
RUNX1 (AML1, CBFA2) is mutated in affected members of families with autosomal dominant thrombocytopenia and platelet dense granule storage pool deficiency. Many of those affected, usually by point mutations in one allele, are predisposed to the development of acute myeloid leukemia (AML) in adult life. The RUNX1 protein complexes with core binding factor beta (CBFB) to form a heterodimeric core binding transcription factor (CBF) that regulates many genes important in hematopoiesis. RUNX1 was first identified as the gene on chromosome 21 that is rearranged by the translocation t(8;21)(q22;q22.12) recurrently found in the leukemic cells of patients with AML. In addition to the t(8;21), RUNX1 is rearranged with one of several partner genes on other chromosomes by somatically acquired translocations associated with hematological malignancies. Point mutations of RUNX1 are also found in sporadic leukemias to reinforce the important position of this gene on the multi-step path to leukemia. In animal models, at least one functional copy of RUNX1 is required to effect definitive embryonic hematopoiesis. Cells expressing dominant-negative mutants of RUNX1 are readily immortalized and transformed, and those RUNX1 mutants which retain CBFB binding ability may possess dominant-negative function. However, in some families there is transmitted one mutated allele of RUNX1 with no dominant-negative function, demonstrating that simple haploinsufficiency of RUNX1 predisposes to AML and also causes a generalized hematopoietic stem cell disorder most recognizable as thrombocytopenia.
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Proteínas de Ligação a DNA/genética , Predisposição Genética para Doença/genética , Leucemia Mieloide Aguda/genética , Mutação/genética , Proteínas Proto-Oncogênicas/genética , Fatores de Transcrição/genética , Animais , Cromossomos Humanos Par 21/genética , Subunidade alfa 2 de Fator de Ligação ao Core , Modelos Animais de Doenças , HumanosRESUMO
BACKGROUND: Myelofibrosis occurs as primary myelofibrosis or as a late occurrence in the evolution of essential thrombocythaemia and polycythaemia vera. It is the rarest of the three classic myeloproliferative neoplasms (MPN). Polyploidy has only rarely been reported in MPN despite the prominent involvement of abnormal megakaryocytes. The use of peripheral blood samples containing increased numbers of haematopoietic progenitors has improved the output from cytogenetic studies in myelofibrosis and together with the use of single nucleotide polymorphism arrays (SNPa) has contributed to an improved knowledge regarding the diverse genetic landscape of this rare disease. RESULTS: Cytogenetic studies performed on a consecutive cohort of 42 patients with primary or post ET/PV myelofibrosis showed an abnormal karyotype in 24 cases and of these, nine showed a polyploid clone. Six of the nine cases showed a tetraploid (4n) subclone, whereas three showed mixed polyploid subclones with both tetraploid and octoploid (4n/8n) cell lines. The abnormal clone evolved from a near diploid karyotype at the initial investigation to a tetraploid karyotype in follow-up cytogenetic analysis in four cases. In total, six of the nine polyploid cases showed gain of 1q material. The remaining three cases showed polyploid metaphases, but with no detectable structural karyotypic rearrangements. Three of the nine cases showed chromosome abnormalities of 6p, either at diagnosis or later acquired. SNPa analysis on eight polyploid cases showed additional changes not previously recognised by karyotype analysis alone, including recurring changes involving 9p, 14q, 17q and 22q. Except for gain of 1q, SNPa findings from the polyploid group compared to eight non-polyploid cases with myelofibrosis found no significant differences in the type of abnormality detected. CONCLUSIONS: The study showed the use of peripheral blood samples to be suitable for standard karyotyping evaluation and DNA based studies. The overall profile of abnormalities found were comparable with that of post-MPN acute myeloid leukaemia or secondary myelodysplastic syndrome and cases in the polyploidy group were associated with features of high risk disease. The above represents the first documented series of polyploid karyotypes in myelofibrosis and shows a high representation of gain of 1q.
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We previously reported molecular karyotype analysis of invasive breast tumour core needle biopsies by comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH) (Walker et al, Genes Chromosomes Cancer, 2008 May;47(5):405-17). That study identified frequently recurring gains and losses involving chromosome bands 8q22 and 8p21, respectively. Moreover, these data highlighted an association between 8q22 gain and typically aggressive grade 3 tumors. Here we validate and extend our previous investigations through FISH analysis of tumor touch imprints prepared from excised breast tumor specimens. Compared to post-surgical tumor excisions, core needle biopsies are known to be histologically less precise when predicting tumor grade. Therefore investigating these chromosomal aberrations in tumor samples that offer more reliable pathological assessment is likely to give a better overall indication of association. A series of 60 breast tumors were screened for genomic copy number changes at 8q22 and 8p21 by dual-color FISH. Results confirm previous findings that 8p loss (39%) and 8q gain (74%) occur frequently in invasive breast cancer. Both absolute quantification of 8q22 gain across the sample cohort, and a separate relative assessment by 8q22:8p21 copy number ratio, showed that the incidence of 8q22 gain significantly increased with grade (p = 0.004, absolute and p = 0.02, relative). In contrast, no association was found between 8p21 loss and tumor grade. These findings support the notion that 8q22 is a region of interest for invasive breast cancer pathogenesis, potentially harboring one or more genes that, when amplified, precipitate the molecular events that define high tumor grade.
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Neoplasias da Mama/diagnóstico , Carcinoma Ductal de Mama/diagnóstico , Cromossomos Humanos Par 8 , Translocação Genética , Idoso , Neoplasias da Mama/genética , Neoplasias da Mama/mortalidade , Neoplasias da Mama/cirurgia , Carcinoma Ductal de Mama/genética , Carcinoma Ductal de Mama/mortalidade , Carcinoma Ductal de Mama/cirurgia , Bandeamento Cromossômico , Variações do Número de Cópias de DNA , Feminino , Humanos , Hibridização in Situ Fluorescente , Cariotipagem , Mastectomia , Pessoa de Meia-Idade , Gradação de Tumores , Análise de SobrevidaRESUMO
Chronic myeloid leukemia (CML) is a clonal myeloproliferative disease caused by recombination between the BCR gene on chromosome 22 and the ABL1 gene on chromosome 9. This rearrangement generates the BCR-ABL1 fusion gene that characterizes leukemic cells in all CML cases. In about 90% of cases, the BCR-ABL1 rearrangement is manifest cytogenetically by the Philadelphia (Ph) chromosome, a derivative of the reciprocal translocation t(9;22)(q34;q11.2). For the remaining cases, recombination may be more complex, involving BCR, ABL1, and genomic sites on one or more other chromosomal regions, or it may occur cryptically within an apparently normal karyotype. Detection of the Ph and associated t(9;22) translocation is a recognized clinical hallmark for CML diagnosis. The disease has a natural multistep pathogenesis, and during chronic phase CML, the t(9;22) or complex variant is usually the sole abnormality. In 60-80% of cases, additional cytogenetic changes appear and often forecast progression to an accelerated disease phase or a terminal blast crisis. Because new frontline therapies such as imatinib specifically target the abnormal protein product of the BCR-ABL1 fusion gene to eliminate BCR-ABL1 positive cells, there is a new reliance on the cytogenetic evaluation of bone marrow cells at diagnosis, then at regular posttreatment intervals. Combined with other parameters, presence or absence of Ph-positive cells in the bone marrow is a powerful early indicator for clinical risk stratification. Cytogenetic changes detected at any stage during treatment, including in the BCR-ABL1-negative cells, may also provide useful prognostic information. Laboratory methods detailed here extend from initial collection of peripheral blood or bone marrow samples through cell culture with or without synchronization, metaphase or interphase harvest, hypotonic treatment and fixation, slide preparation for G-banding or fluorescent in situ hybridization (FISH), and final interpretation.
Assuntos
Análise Citogenética/métodos , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Leucemia Mielogênica Crônica BCR-ABL Positiva/terapia , Buffy Coat/metabolismo , Células da Medula Óssea/metabolismo , Células da Medula Óssea/patologia , Técnicas de Cultura de Células , Separação Celular , Bandeamento Cromossômico , Feminino , Humanos , Hibridização in Situ Fluorescente , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Contagem de Leucócitos , Masculino , Manejo de Espécimes , Fatores de TempoRESUMO
The RUNX1 (AML1, CBFA2) gene is a member of the runt transcription factor family, responsible for DNA binding and heterodimerization of other non-DNA binding transcription factors. RUNX1 plays an important part in regulating haematopoiesis and it is frequently disrupted by illegitimate somatic recombination in both acute myeloid and lymphoblastic leukaemia. Germline mutations of RUNX1 have also recently been described and are dominantly associated with inherited leukaemic conditions. We have identified a unique point mutation of the RUNX1 gene (A107P) in members of a family with autosomal dominant inheritance of thrombocytopenia. One member has developed acute myeloid leukaemia (AML).