what's the time required for fixing solid tumor in 10% formaldehyde for IHC buffer embedded tissue?
By Shrouk El khamisy · Benha UniversityI would like to support Maria Soares's recommendation as there is evidence for it in relation to routinely tested breast cancer biomarkers including ... [more]I would like to support Maria Soares's recommendation as there is evidence for it in relation to routinely tested breast cancer biomarkers including ER. PR. and HER2. As for the size of the tissue being it is important to attempt to slice larger tissue specimens down to to 1x1x0.5cm portions. It is important to regularly ascertain that any stored buffered formalin is at neutral or slightly above pH 7, before using itFollowing
Does anyone work with tamoxifen resistant MCF-7 breast cancer cell line? Has anyone checked the action of tamoxifen to that resistant cell?
By Nipin Sp · Konkuk UniversityYou may wish to follow up the work of my colleagues Dr Julia and Prof Bob Nicholson in Cardiff using the following weblink of their latest work on lon... [more]You may wish to follow up the work of my colleagues Dr Julia and Prof Bob Nicholson in Cardiff using the following weblink of their latest work on long term cultures of tamoxifen resistant MCF-7 cell lines http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3393679/Following
How can circulating tumor cells exist at all?
By Christer Ericsson · Karolinska InstituteI have a lateral thought to offer which could help to accommodate the differing views expressed above. There is evidence to suggest that CTCs may be d... [more]I have a lateral thought to offer which could help to accommodate the differing views expressed above. There is evidence to suggest that CTCs may be derived from bone marrow spread of the original tumour. This is implied from the finding that patients with CTCs have a great propensity to develop bone metastases. An inference one can derive from this is that the initial micro-metastatic spread to the bone marrow ( which speculated to occur in ~30% of breast cancer patients) if it survives and becomes established as a nidus of viable tumour deposit, it may remain latent until a clone evolves with characteristics of bone marrow derived cells (i.e. blood cells). Such cells may have the capacity to survive when travelling through the blood circulation as well as circulate through the capillary bed like ordinary blood cells.Following
Answer added in Protein Biochemistry13 Can testosterone be immunodetected in formalin-fixed testes?By Breno Caneguim · Universidade Federal de São PauloDespite my reservation I have spotted one paper from Croatia describing IHC localisation of testosterone in Leydig cells at ultrastructural level. The... [more]Despite my reservation I have spotted one paper from Croatia describing IHC localisation of testosterone in Leydig cells at ultrastructural level. The following link relates to an abstract of the paper http://journals.cambridge.org/action/displayFulltext?type=1&fid=174502&jid=MAM&volumeId=9&issueId=S03&aid=174501Following
Answer added in Protein Biochemistry13 Can testosterone be immunodetected in formalin-fixed testes?By Breno Caneguim · Universidade Federal de São PauloI feel that small diffusible molecules like testosterone will not be retained to any degree even in optimally fixed tissue. In addition there is the d... [more]I feel that small diffusible molecules like testosterone will not be retained to any degree even in optimally fixed tissue. In addition there is the danger of any diffused teststerone molecules giving false positive results outside the site of normal synthesis and storageFollowing
Article: Detection of breast cancer metastasis in sentinel lymph nodes using intra-operative real time GeneSearch™ BLN Assay in the operating room: results of the Cardiff studyRobert E. Mansel, Amit Goyal, Anthony Douglas-Jones, Victoria Woods, Sumit Goyal, Ian Monypenny, Helen Sweetland, Robert G. Newcombe, Bharat Jasani[show abstract] [hide abstract]
ABSTRACT: Background Intra-operative assessment is not routinely performed in the UK due to poor sensitivity of available methods and overburdened pathology resources. We conducted a prospective clinical feasibility study of the GeneSearch™ Breast Lymph Node (BLN) Assay (Veridex, LLC, Warren, NJ) to confirm its potential usefulness within the UK healthcare system. Methods In the assay 50% of the lymph node was processed to detect the presence of cytokeratin-19 and mammaglobin mRNA. The assay was calibrated to detect metastases >0.2mm. Assay results were compared to H&E performed on each face of ~2mm alternating node slabs and 3 additional sections cut at ~150μm interval from each face of the node slab. Results 124 sentinel lymph nodes were removed from 82 breast cancer patients. The assay correctly identified all 6 patients with sentinel node macrometastases (>2.0mm), and 2 of 3 patients with sentinel node micrometastases (0.2–2.0mm). Sentinel lymph nodes in 4 patients were assay positive but histology negative. Two of these four patients had isolated tumor cells seen by histology. The overall concordance with histology was 93.9% (77/82), with sensitivity of 88.9% (8/9, 95% CI 56.5–98%), specificity of 94.6% (69/73, 95% CI 86.7–97.8%), positive predictive value of 66.7% (8/12, 95% CI 39.1–86.2%) and negative predictive value of 98.6% (69/70, 95% CI 92.3–99.7%). The assay was performed in a median time of 32min (range 26–69min). Conclusion Intra-operative assessment of sentinel lymph node can be performed rapidly and accurately using the GeneSearch™ BLN Assay.Breast Cancer Research and Treatment 04/2012; 115(3):595-600. · 4.43 Impact Factor
Article: HER2 gene amplification in breast cancer: a rogues' gallery of challenging diagnostic cases: UKNEQAS interpretation guidelines and research recommendations.Jane Starczynski, Neil Atkey, Yvonne Connelly, Tony O'Grady, Fiona M Campbell, Silvana di Palma, Peter Wencyk, Bharat Jasani, Michael Gandy, J M S Bartlett[show abstract] [hide abstract]
ABSTRACT: International and national guidelines highlight the importance of accuracy, reproducibility, and quality control of in situ hybridization (ISH) methods for testing breast carcinomas. However, few guidelines cover the reporting of ISH cases with "unusual" signal patterns, including, eg, heterogeneity and loss of chromosome enumeration probe or gene signals. These cases are, in fact, relatively frequent, and there is a need for developing evidence- or consensus-based reporting guidelines to ensure consistency of treatment. Following an audit of cases from a single center (including >1,700 cases) we show that approximately 10% of ISH results reflect unusual signal patterns. We illustrate the most common of these patterns and provide reporting guidelines for diagnosticians and recommendations for future research. Our goal is to ensure that in the future such "rogues" are reported in a consistent manner that, ultimately, will be supported by molecular and biochemical evidence.American Journal of Clinical Pathology 04/2012; 137(4):595-605. · 2.60 Impact Factor
Bharat Jasani, Allen Gibbs[show abstract] [hide abstract]
ABSTRACT: Despite asbestos being identified as the single most important cause of malignant mesothelioma, the tumor is known to occur in only 10% to 20% of heavily exposed individuals. In addition, about 20% of the patients have no history of asbestos exposure even after detailed assessment. Therefore, there has been speculation for some time that asbestos alone may not be sufficient to cause mesothelioma and that other factors may be involved either as cocarcinogens or as independent mechanisms of cancer causation. To give a brief review of nonasbestos fiber erionite and therapeutic radiation as 2 established examples of asbestos-independent mechanisms, of the potential emerging role of man-made fibers such as carbon nanotubes, and of polyoma virus SV40 (simian virus 40) as a potential example of the cocarcinogenic mode of involvement. Relevant recent literature has been surveyed to portray and provide the evidence in favor of the examples. Erionite has emerged as the most important example of nonasbestos-mediated cause of mesothelioma in regions such as Turkey where exposure to this type of fiber is highly prevalent. Recently, the polyoma virus SV40 has been unexpectedly discovered as an effective cocarcinogen of asbestos in the causation of animal mesothelioma, though despite considerable research, its potential role in human mesothelioma remains unproven.Archives of pathology & laboratory medicine 03/2012; 136(3):262-7. · 2.58 Impact Factor
Article: HER2 testing in the UK: recommendations for breast and gastric in-situ hybridisation methods.J M S Bartlett, J Starczynski, Neil Atkey, E Kay, A O'Grady, Michael Gandy, Merdol Ibrahim, Bharat Jasani, I O Ellis, S E Pinder, R A Walker[show abstract] [hide abstract]
ABSTRACT: These guidelines supplement existing guidelines on HER2 testing by immunohistochemistry and in-situ hybridisation(ISH) methods in the UK. They provide a specific focus on aspects of guidance relevant to HER2 ISH testing methods, both fluorescent and chromogenic. They are formulated to give advice on methodology, interpretation and quality control for ISH-based testing of HER2 status in common tumour types, including both breast and gastric tumours. The aim is to ensure that all ISH-based testing is accurate, reliable and timely.Journal of clinical pathology 06/2011; 64(8):649-53. · 2.43 Impact Factor
Article: Addition of cetuximab to oxaliplatin-based first-line combination chemotherapy for treatment of advanced colorectal cancer: results of the randomised phase 3 MRC COIN trial.Timothy S Maughan, Richard A Adams, Christopher G Smith, Angela M Meade, Matthew T Seymour, Richard H Wilson, Shelley Idziaszczyk, Rebecca Harris, David Fisher, Sarah L Kenny, [......], Jenna K Mitchell, Ayman Madi, Bharat Jasani, Michelle D James, John Bridgewater, M John Kennedy, Bart Claes, Diether Lambrechts, Richard Kaplan, Jeremy P Cheadle[show abstract] [hide abstract]
ABSTRACT: In the Medical Research Council (MRC) COIN trial, the epidermal growth factor receptor (EGFR)-targeted antibody cetuximab was added to standard chemotherapy in first-line treatment of advanced colorectal cancer with the aim of assessing effect on overall survival. In this randomised controlled trial, patients who were fit for but had not received previous chemotherapy for advanced colorectal cancer were randomly assigned to oxaliplatin and fluoropyrimidine chemotherapy (arm A), the same combination plus cetuximab (arm B), or intermittent chemotherapy (arm C). The choice of fluoropyrimidine therapy (capecitabine or infused fluouroracil plus leucovorin) was decided before randomisation. Randomisation was done centrally (via telephone) by the MRC Clinical Trials Unit using minimisation. Treatment allocation was not masked. The comparison of arms A and C is described in a companion paper. Here, we present the comparison of arm A and B, for which the primary outcome was overall survival in patients with KRAS wild-type tumours. Analysis was by intention to treat. Further analyses with respect to NRAS, BRAF, and EGFR status were done. The trial is registered, ISRCTN27286448. 1630 patients were randomly assigned to treatment groups (815 to standard therapy and 815 to addition of cetuximab). Tumour samples from 1316 (81%) patients were used for somatic molecular analyses; 565 (43%) had KRAS mutations. In patients with KRAS wild-type tumours (arm A, n=367; arm B, n=362), overall survival did not differ between treatment groups (median survival 17·9 months [IQR 10·3-29·2] in the control group vs 17·0 months [9·4-30·1] in the cetuximab group; HR 1·04, 95% CI 0·87-1·23, p=0·67). Similarly, there was no effect on progression-free survival (8·6 months [IQR 5·0-12·5] in the control group vs 8·6 months [5·1-13·8] in the cetuximab group; HR 0·96, 0·82-1·12, p=0·60). Overall response rate increased from 57% (n=209) with chemotherapy alone to 64% (n=232) with addition of cetuximab (p=0·049). Grade 3 and higher skin and gastrointestinal toxic effects were increased with cetuximab (14 vs 114 and 67 vs 97 patients in the control group vs the cetuximab group with KRAS wild-type tumours, respectively). Overall survival differs by somatic mutation status irrespective of treatment received: BRAF mutant, 8·8 months (IQR 4·5-27·4); KRAS mutant, 14·4 months (8·5-24·0); all wild-type, 20·1 months (11·5-31·7). This trial has not confirmed a benefit of addition of cetuximab to oxaliplatin-based chemotherapy in first-line treatment of patients with advanced colorectal cancer. Cetuximab increases response rate, with no evidence of benefit in progression-free or overall survival in KRAS wild-type patients or even in patients selected by additional mutational analysis of their tumours. The use of cetuximab in combination with oxaliplatin and capecitabine in first-line chemotherapy in patients with widespread metastases cannot be recommended. Cancer Research UK, Cancer Research Wales, UK Medical Research Council, Merck KGgA.The Lancet 06/2011; 377(9783):2103-14. · 38.28 Impact Factor