Analytical Cellular Pathology - Volume 34, issue 1-2

Authors: Cohen, Stanley

Article Type: Research Article

DOI: 10.3233/ACP-2011-0007

Citation: Analytical Cellular Pathology, vol. 34, no. 1-2, pp. 1-2, 2011

Authors: Chen, Xiaodong | Zheng, Bin | Liu, Hong

Article Type: Research Article

Abstract: The conventional optical microscope has been the primary tool in assisting pathological examinations. The modern digital pathology combines the power of microscopy, electronic detection, and computerized analysis. It enables cellular-, molecular-, and genetic-imaging at high efficiency and accuracy to facilitate clinical screening and diagnosis. This paper first reviews the fundamental concepts of microscopic imaging and introduces the technical features and associated clinical applications of optical microscopes, electron microscopes, scanning tunnel microscopes, and fluorescence microscopes. The interface of microscopy with digital image acquisition methods is discussed. The recent developments and future perspectives of contemporary microscopic imaging techniques such as three-dimensional and …in vivo imaging are analyzed for their clinical potentials. Show more

DOI: 10.3233/ACP-2011-0006

Citation: Analytical Cellular Pathology, vol. 34, no. 1-2, pp. 5-18, 2011

Authors: Timme, Sylvia | Schmitt, Eberhard | Stein, Stefan | Schwarz-Finsterle, Jutta | Wagner, Jenny | Walch, Axel | Werner, Martin | Hausmann, Michael | Wiech, Thorsten

Article Type: Research Article

Abstract: Cell type specific radial positioning of chromosome territories (CTs) and their sub-domains in the interphase seem to have functional relevance in non-neoplastic human nuclei, while much less is known about nuclear architecture in carcinoma cells and its development during tumor progression. We analyzed the 3D-architecture of the chromosome 8 territory (CT8) in carcinoma and corresponding non-neoplastic ductal pancreatic epithelium. Fluorescence-in-situ-hybridization (FISH) with whole chromosome painting (WCP) probes on sections from formalin-fixed, paraffin wax-embedded tissues from six patients with ductal adenocarcinoma of the pancreas was used. Radial positions and shape parameters of CT8 were analyzed by 3D-microscopy. None of the parameters …showed significant inter-individual changes. CT8 was localized in the nuclear periphery in carcinoma cells and normal ductal epithelial cells. Normalized volume and surface of CT8 did not differ significantly. In contrast, the normalized roundness was significantly lower in carcinoma cells, implying an elongation of neoplastic cell nuclei. Unexpectedly, radial positioning of CT8, a dominant parameter of nuclear architecture, did not change significantly when comparing neoplastic with non-neoplastic cells. A significant deformation of CT8, however, accompanies nuclear atypia of carcinoma cells. This decreased roundness of CTs may reflect the genomic and transcriptional alterations in carcinoma. Show more

Keywords: Pancreatic adenocarcinoma, nuclear architecture, chromosome 8 territory, radial positioning, shape of chromosome 8 territory

DOI: 10.3233/ACP-2011-0004

Citation: Analytical Cellular Pathology, vol. 34, no. 1-2, pp. 21-33, 2011

Authors: Katz, Elad | Verleyen, Wim | Blackmore, Colin G. | Edward, Michael | Smith, V. Anne | Harrison, David J.

Article Type: Research Article

Abstract: Tumour cells employ a variety of mechanisms to invade their environment and to form metastases. An important property is the ability of tumour cells to transition between individual cell invasive mode and collective mode. The switch from collective to individual cell invasion in the breast was shown recently to determine site of subsequent metastasis. Previous studies have suggested a range of invasion modes from single cells to large clusters. Here, we use a novel image analysis method to quantify and categorise invasion. We have developed a process using automated imaging for data collection, unsupervised morphological examination of breast cancer invasion …using cognition network technology (CNT) to determine how many patterns of invasion can be reliably discriminated. We used Bayesian network analysis to probabilistically connect morphological variables and therefore determine that two categories of invasion are clearly distinct from one another. The Bayesian network separated individual and collective invading cell groups based on the morphological measurements, with the level of cell-cell contact the most discriminating morphological feature. Smaller invading groups were typified by smoother cellular surfaces than those invading collectively in larger groups. Interestingly, elongation was evident in all invading cell groups and was not a specific feature of single cell invasion as a surrogate of epithelial-mesenchymal transition. In conclusion, the combination of cognition network technology and Bayesian network analysis provides an insight into morphological variables associated with transition of cancer cells between invasion modes. We show that only two morphologically distinct modes of invasion exist. Show more

Keywords: Invasion, imaging, breast cancer, Bayesian networks, EMT

DOI: 10.3233/ACP-2011-0003

Citation: Analytical Cellular Pathology, vol. 34, no. 1-2, pp. 35-48, 2011

Authors: Pelekanou, Vassiliki | Notas, George | Theodoropoulou, Katerina | Kampa, Marilena | Takos, Dimitrios | Alexaki, Vassilia-Ismini | Radojicic, Jelena | Sofras, Frank | Tsapis, Andreas | Stathopoulos, Efstathios N. | Castanas, Elias

Article Type: Research Article

Abstract: In advanced renal cell carcinoma (RCC), surgery combined with systemic chemotherapy and immunotherapy have had limited effectiveness. Therapeutic modalities targeting VEGF, PDGF, and c-kit using tyrosine kinase inhibitors and m-TOR using specific biologic factors are in development. Therapeutic approaches targeting TNF-alpha have shown limited efficacy, while anti-TRAIL (TNFSF10) antibodies have shown enhanced activity. The presence and potential significance of other members of the TNFSF has not been investigated. Here, we assayed the TNFSF members APRIL, BAFF, TWEAK and their receptors (BCMA, TACI, BAFFR, Fn14) in 86 conventional type clear cell RCC, using immunohistochemistry and correlated our findings with histological data …and, in a limited series, follow-up of patients. We observed a differential expression of these TNFSF ligands and receptors in cancerous and non-cancerous structures. BAFF was found in all RCC; APRIL expression is associated with an aggressive phenotype, correlating negatively with patients' disease-free survival, while TWEAK and its receptor Fn14 are heterogeneously expressed, correlating negatively with the grade and survival of RCC patients. This is the first study, presenting together the TNFSF members APRIL, BAFF, TWEAK and their receptors in different areas of normal renal tissue and RCC, suggesting a potential role of these TNFSF members in renal tumor biology. Show more

Keywords: Renal cell carcinoma conventional type, tumor necrosis factor superfamily members (BAFF, APRIL, TWEAK), tumor necrosis factor receptor superfamily members (BCMA, TACI, TWEAK, Fn14)

DOI: 10.3233/ACP-2011-0001

Citation: Analytical Cellular Pathology, vol. 34, no. 1-2, pp. 49-60, 2011

Authors: Richman, Susan D. | Chambers, Philip | Seymour, Matthew T. | Daly, Catherine | Grant, Sophie | Hemmings, Gemma | Quirke, Philip

Article Type: Research Article

Abstract: KRAS mutation status is established as a predictive biomarker of benefit from anti-EGFr therapies. Mutations are normally assessed using DNA extracted from one formalin-fixed, paraffin-embedded (FFPE) tumor block. We assessed heterogeneity of KRAS and BRAF mutation status intra-tumorally (multiple blocks from the same primary tumor). We also investigated the utility and efficiency of genotyping a ‘DNA cocktail’ prepared from multiple blocks. We studied 68 consenting patients in two randomized clinical trials. DNA was extracted, from ≥2 primary tumor FFPE blocks per patient. DNA was genotyped by pyrosequencing for KRAS codons 12, 13 and 61 and BRAF codon 600. In patients …with heterogeneous mutation status, DNA cocktails were prepared and genotyped. Among 69 primary tumors in 68 patients, 7 (10.1%) showed intratumoral heterogeneity; 5 (7.2%) at KRAS codons 12, 13 and 2 (2.9%) at BRAF codon 600. In patients displaying heterogeneity, the relevant KRAS or BRAF mutation was also identified in ‘DNA cocktail’ samples when including DNA from mutant and wild-type blocks. Heterogeneity is uncommon but not insignificant. Testing DNA from a single block will wrongly assign wild-type status to 10% patients. Testing more than one block, or preferably preparation of a ‘DNA cocktail’ from two or more tumor blocks, improves mutation detection at minimal extra cost. Show more

Keywords: KRAS, BRAF, heterogeneity, colorectal cancer

DOI: 10.3233/ACP-2011-0005

Citation: Analytical Cellular Pathology, vol. 34, no. 1-2, pp. 61-66, 2011

Authors: Ammar, Aula | Mohammed, Rabab A.A. | Salmi, Marko | Pepper, Michael | Paish, Emma C. | Ellis, Ian O. | Martin, Stewart G.

Article Type: Research Article

Abstract: Background: Mechanisms regulating breast cancer lymph node metastasis are unclear. Staining of CLEVER-1 (common lymphatic endothelial and vascular endothelial receptor-1) in human breast tumors was used, along with in vitro techniques, to assess involvement in the metastatic process. Methods: 148 sections of primary invasive breast cancers, with 10 yr follow-up, were stained with anti-CLEVER-1. Leukocyte infiltration was assessed, along with involvement of specific subpopulations by staining with CD83 (mature dendritic cells, mDC), CD209 (immature DC, iDC) and CD68 (macrophage, M�). In vitro expression of CLEVER-1 on lymphatic (LEC) and blood endothelial cells (BEC) was examined by flow cytometry. Results: In …vitro results showed that although both endothelial cell types express CLEVER-1, surface expression was only evident on LEC. In tumour sections CLEVER-1 was expressed in blood vessels (BV, 61.4% of samples), lymphatic vessels (LV, 18.2% of samples) and in M�/DCs (82.4% of samples). However, only CLEVER-1 expression in LV was associated with LN metastasis (p = 0.027) and with M� indices (p = 0.021). Although LV CLEVER-1 was associated with LN positivity there was no significant correlation with recurrence or overall survival, BV CLEVER-1 expression was, however, associated with increased risk of recurrence (p = 0.049). The density of inflammatory infiltrate correlated with CLEVER-1 expression in BV (p < 0.001) and LV (p = 0.004). Conclusions: The associations between CLEVER-1 expression on endothelial vessels and macrophage/leukocyte infiltration is suggestive of its regulation by inflammatory conditions in breast cancer, most likely by macrophage-associated cytokines. Its upregulation on LV, related surface expression, and association with LN metastasis suggest that it may be an important mediator of tumor cell metastasis to LN. Show more

Keywords: Breast cancer, lymph node metastasis, CLEVER-1, lymphatic, tumor cell adhesion

DOI: 10.3233/ACP-2011-0002

Citation: Analytical Cellular Pathology, vol. 34, no. 1-2, pp. 67-78, 2011