134 research outputs found
Fluorescence in situ hybridisation BRAF-mutation analysis on pre-stained smears from thyroid fine-needle aspiration cytology
No full textMaster i biomedisinBackground: PTC is a differentiated thyroid carcinoma and the most common (70-80%) among all thyroid
cancers. BRAFV600E-mutation is the most prevalent in PTC and a highly sensitive marker. Ultrasoundguided
fine-needle aspiration cytology (FNAC) as a fundamental tool is highly sensitive for diagnosing
thyroid nodules. However, ~25 % are of indeterminate lesions. Hence, the purpose is to develop a FISH
BRAFV600E-mutation analysis on pre-operative thyroid FNAC smears stained with MGG or DiffQuick to
eliminate such indeterminate lesions.
Methods: MGG or DQ-stained FNAC smears of pre-operative thyroid nodules histologically verified as PTC
were used to work out a FISH BRAF-mutation analysis. ~50 smears were tested, where different
parameters on the FISH assay were combined for an optimal protocol. After protocol modification, 28
smears from 14 individuals, where 14 smears were stained with a BRAF break-apart translocation probe
and the rest 14 with BRAF amplification probe, were tested.
Results: Optimal results with a combination of a TE buffer (pH 9), pepsin digestion and post-fixation of
smears in formalin 10 %, were obtained. Signals of BRAF break-apart probe and amplification probe were
detected using a Zeiss confocal microscope.
Conclusion: A modified FISH method on air-dried and Giemsa/DQ stained smears from thyroid FNAC is
suitable on areas with least bloodstain on smears avoiding an autofluorescence. A combination of
pretreatment buffer TE at pH 9, pepsin digestion and a 5 mins post-fixation in formalin 10 % are highly
beneficial steps in developing this method.publishedVersio
Fine-needle aspiration cytology of extra mammary metastatic lesions in the breast: A retrospective study of 36 cases diagnosed during 18 years
No full textBackground:Metastatic tumors in the breast require treatment according to origin and type of tumor. It is important to recognize these lesions in fine-needle aspiration cytology (FNAC) in order to avoid unnecessary mastectomy or non-relevant chemotherapy. The aim of this study was to evaluate the cytological features of metastatic tumors and possible criteria that could alert us as to the possibility of a metastasis from an extra mammary malignancy.Methods:The material included 36 confirmed or suspected metastases in the breast registered in the pathology files at Oslo University Hospital, Ulleval, during 1990–2007. There were a total of 6,325 cases of malignant breast FNAC, representing 30 men and 6,295 women. Smears were evaluated for the amount of material, presence or absence of myoepithelial cells, microcalcifications, mitoses and necrotic material. All carcinomas were graded.Results:There were seven men (7/30 = 23.3%) and 29 women (29/6,295 = 0.46%). The primary tumor was known in 22 cases (22/36 = 61.1%). No other primary tumor was known and metastatic lesion was not initially suspected in 14 cases (14/36 = 38.9%). The most common origin was lung (15/36 = 41.7%). In five cases (5/36 = 13.9%), the origin remained uncertain.Conclusions:Metastases from extra mammary sites are (relatively) common in males (23.3%). In women, metastatic lesions are rare (0.46%). A large proportion of them (88%) are high-grade adenocarcinomas and poorly differentiated carcinomas that may resemble grade 3 ductal carcinomas. Unusual clinical and/or radiological presentation in combination with high-grade malignant cells should alert us to consider the possibility of a metastasis.</jats:sec
Assessing invasion criteria in fine needle aspirates from breast carcinoma diagnosed as dcis or invasive carcinoma: can we identify an invasive component in addition to dcis
No full textObjective: To evaluate invasion criteria in fine needle aspiration cytology (FNAC) of histologically diagnosed breast ductal carcinoma in situ (DCIS) and invasive carcinoma and to evaluate their usefulness in identifying an invasive component in addition to DCIS.
Study Design: The material consisted of 331 smears diagnosed as suspicious for or consistent with DCIS and in which histology had shown either DCIS or invasive ductal carcinoma. All smears were reevaluated for the following invasion criteria: invasion of fat or fibrous tissue fragments, fibroblast proliferation, cell-poor elastoid tissue fragments, tubular structures and intracytoplasmic vacuoles.
Results: All invasion criteria except cytoplasmic vacuoles correlated with invasiveness, but none of them were found exclusively in invasive lesions. Pseudoinvasion in fibrous or fatty tissue fragments were found in 8 cases of histologic pure DCIS. One DCIS (0.4%) revealed ?2 invasion features as well as 22 invasive carcinomas (20.7%), representing 7.4% of all cases.
Conclusion: Using established invasion criteria, practically no pure DCIS lesion will be diagnosed as invasive on FNAC, but one will identify only a subset of cases harboring an invasive componen
Expression of the pS2 protein and correlation with estrogen receptor status in fine‐needle aspirates from breast carcinomas
No full textDetermination of HER-2 status on FNAC material from breast carcinomas using<i>in situ</i>hybridization with dual chromogen visualization with silver enhancement (dual SISH)
No full textDuring the last years, HER-2 status kits and protocols for chromogen visualization of hybridization signals have come on the market. The first generation using chromogen visualization used single color probes. The second generation, now emerging on the market, uses dual chromogen visualization. The aim of this study has been to test a new dual color chromogen kit (Ventana INFORM HER2 Dual Colour ISH Roche®) and compare the results with our in-house method(s). The material consisted primarily of cytological material from invasive breast carcinomas in 49 women. Dual SISH was done on all 49 cytological and histological specimens. The histological specimens were treated according to the manufacturer’s recommendations. The procedure was modified in several steps in order to adapt it to the cytological material. Hybridization failed in two cytological specimens. Dual SISH showed concordant results on cytological and histological material as to amplified/not amplified. The included cases had the same HER-2 expression in the invasive and thein situcomponents on histology. Four IDC showed HER-2 amplification (8.5%). Polysomy was found in two cases. All dual SISH results except for one concurred with the results of the in-house method(s) (1/47=2.1%). The dual SISH is suitable for cytological examination of HER-2 status. The protocol must be optimized for cytological material.</jats:p
Abstract LB-027: The EP300-G211S mutation is highly associated with a low mutational burden in triple-negative breast cancer patients
No full textAbstract
Introduction. Next-generation sequencing (NGS) technologies offer the possibility to assess multiple genes for somatic mutations and may elucidate the driver genetic variations involved in carcinogenesis and disease progression. Among the different subtypes of breast cancer, the triple-negative subgroup (TNBC) is characterized by poor prognosis and a lack of reliable tumor markers. In particular, molecular profiling and the clinical observation that some patients with TNBC relapse early after surgery and show only poor responses to established therapies while others do not relapse at all, indicate the existence of major subgroups within the TNBC cohort. The aim of this study was to characterize somatic variants in breast cancer tumors obtained from a cohort of 147 Norwegian patients using a NGS panel with special emphasis on the TNBC subtype (57% of the patients).
Methods. Genomic DNA was extracted from paraffin embedded formalin fixed (FFPE) tissue obtained from 147 consecutive patients diagnosed with a primary BC at our hospital. The DNA samples were analyzed by next-generation sequencing (NGS) using Human Breast Cancer GeneRead DNAseq Targeted Panel V2 (Qiagen). The panel consists of a collection of PCR primers for targeted enrichment of the coding region of 44 genes commonly mutated in breast cancer. Target enrichment and library construction was performed according to the GeneReader workflow (Qiagen) and paired end sequencing was performed on a NextSeq 500 sequencer (Illumina) running 2 x 150 bp chemistry Version 2. Data analysis including alignment to the reference genome hg19 and variant calling was performed using Qiagen’s online Ingenuity Variant analysis.
Results. The ingenuity variant analysis classified the somatic mutations according to their clinical significance into four groups: pathogenic, likely pathogenic, benign and likely benign (as defined by the American College of Medical Genetics and Genomics). Focusing on the TNBC cases (n = 84), we observed that patients could be separated in two major subgroups, those with a combination of several mutations in pivotal genes (TP53, BRCA1/2, RB1, RET, PIK3CA etc.) and others, without any mutations in typical breast cancer related genes. Interestingly, the majority of tumors presenting with extremely low mutation rates were found to harbor a specific mutation in the EP300 gene (G211S). In fact, only 3 out of 29 TNBC carrying EP300-G211S had other pathogenic mutations. Breast cancer specific survival was significantly improved in EP300-G211S mut. positive patients.
Conclusion. Next generation sequencing of human triple-negative breast cancer samples suggests EP300 to be a pivotal player in breast cancer biology, potentially reflecting the overall mutational burden. These findings should be further investigated as the mutational status of EP300 might have significant implications for clinical decision making.
Note: This abstract was not presented at the meeting.
Citation Format: Vahid Bemanian, Torill Sauer, Joel Touma, Katja M. Vetvik, John C. Noone, Vessela N. Kristensen, Ida R. Bukholm, Jürgen Geisler. The EP300-G211S mutation is highly associated with a low mutational burden in triple-negative breast cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-027. doi:10.1158/1538-7445.AM2017-LB-027</jats:p
Reflections on the Narrative Research Approach
No full textIn her reflections on the narrative research approach, the author starts by placing narrative research within the framework of sociocultural theory, where the challenge for the researcher is to examine and understand how human actions are related to the social context in which they occur and how and where they occur through growth. The author argues that the narrative as a unit of analysis provides the means for doing this. She then presents some of the basic premises of narrative research before she reflects on the process of narrative inquiry and addresses the issue of the “true” narrative. Throughout the article, the author refers to educational research and in the concluding section argues that the results of narrative research can be used as thought-provoking tools within the field of teacher education
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