Jednodnevna dijagnostika

At the University Hospital for Tumors, University Hospital Center Sestre milosrdnice about 700 breast cancers and about 150 benign changes of the breast are operated annually. After mammography and ultrasound, basic distinction between benign and malignant is achieved by fine needle (FNAC aspiration) under visual control of various radiological methods, mainly ultrasound (US). Importance of early cancer diagnosis and planned treatment action achieve the best outcomes. In order to bridge the time gap between different diagnostics and reduce psychological stress, we introduced the concept of one-day clinic into our outpatient practice. Service comprises triple diagnostic test when a lesion is suspected along with treatment plan according to the diagnosis in 8 hour time frame. The service has been operating for the past two years and has had positive response from both staff and patients.U Klinici za tumore Kliničkog bolničkog centra Sestre milosrdnice, godišnje se operira oko 700 karcinoma dojke i oko 150 benignih lezija dojke. Nakon mamografije i ultrazvuka, razlikovanje malignih od benignih tumora se postiže punkcijom pod kontrolom različitih radioloških metoda, prvenstveno ultrazvuka. Važnost ranog otkrivanja raka i planiranje liječenja daju najbolje rezultate. Kako bi se premostili vremenski periodi između različitih dijagnostičkih postupaka i reducirao stres koji nastaje iščekivanjem rezultata, uveli smo koncept jednodnevne dijagnostike. Usluga se sastoji od trojnog dijagnostičkog testa u slučaju sumnje na tumor i plana liječenja u vremenskom okviru od 8 sati. Jednodnevna dijagnostika djeluje zadnje dvije godine i izazvala je pozitivne reakcije kod pacijenata i kod osoblja

Jednodnevna dijagnostika

At the University Hospital for Tumors, University Hospital Center Sestre milosrdnice about 700 breast cancers and about 150 benign changes of the breast are operated annually. After mammography and ultrasound, basic distinction between benign and malignant is achieved by fine needle (FNAC aspiration) under visual control of various radiological methods, mainly ultrasound (US). Importance of early cancer diagnosis and planned treatment action achieve the best outcomes. In order to bridge the time gap between different diagnostics and reduce psychological stress, we introduced the concept of one-day clinic into our outpatient practice. Service comprises triple diagnostic test when a lesion is suspected along with treatment plan according to the diagnosis in 8 hour time frame. The service has been operating for the past two years and has had positive response from both staff and patients.U Klinici za tumore Kliničkog bolničkog centra Sestre milosrdnice, godišnje se operira oko 700 karcinoma dojke i oko 150 benignih lezija dojke. Nakon mamografije i ultrazvuka, razlikovanje malignih od benignih tumora se postiže punkcijom pod kontrolom različitih radioloških metoda, prvenstveno ultrazvuka. Važnost ranog otkrivanja raka i planiranje liječenja daju najbolje rezultate. Kako bi se premostili vremenski periodi između različitih dijagnostičkih postupaka i reducirao stres koji nastaje iščekivanjem rezultata, uveli smo koncept jednodnevne dijagnostike. Usluga se sastoji od trojnog dijagnostičkog testa u slučaju sumnje na tumor i plana liječenja u vremenskom okviru od 8 sati. Jednodnevna dijagnostika djeluje zadnje dvije godine i izazvala je pozitivne reakcije kod pacijenata i kod osoblja

Demystifying molecular techniques in cytopathology practice

The last decade was stimulating with the introduction of new molecular techniques to be applied in pathology laboratories. Accordingly, cytology was also benefited with the innovations emerged from this new era. Molecular cytopathology (MCP) can be defined as molecular studies applied on all types of cytological specimens, namely gynaecology cytology, exfoliative non-gyn cytology and fine needle aspirates. The development of a huge amount of new ancillary techniques has paralleled the emergence of clinical cytology as a major diagnostic speciality. Clinical applications of these techniques have been growing in the last decade. The widespread acceptance of liquid-based systems in gynaecological cytology is a paramount episode which re-draws the relation between cells and molecules. The stretched use of approaches, morphology and molecular biology, in HPV-induced lesions settings, e.g., revealed a potential to optimize, in one single brushed sample, diagnosis and research. Cytology samples from serous effusions, pulmonary tree, bladder urine, and aspirations, among others, are now likely to be studied by different molecular techniques for helping in diagnosis, prognosis, or even to assess therapeutic targets. In this review, we highlight the main results already published concerning the application of molecular techniques in different fields of cytopathology and discuss their application

Regulation of CD4+NKG2D+ Th1 cells in patients with metastatic melanoma treated with sorafenib : role of IL-15Rα and NKG2D triggering

Beyond cancer-cell intrinsic factors, the immune status of the host has a prognostic impact on patients with cancer and influences the effects of conventional chemotherapies. Metastatic melanoma is intrinsically immunogenic, thereby facilitating the search for immune biomarkers of clinical responses to cytotoxic agents. Here, we show that a multi-tyrosine kinase inhibitor, sorafenib, upregulates interleukin (IL)-15Rα in vitro and in vivo in patients with melanoma, and in conjunction with natural killer (NK) group 2D (NKG2D) ligands, contributes to the Th1 polarization and accumulation of peripheral CD4+NKG2D+ T cells. Hence, the increase of blood CD4+NKG2D+ T cells after two cycles of sorafenib (combined with temozolomide) was associated with prolonged survival in a prospective phase I/II trial enrolling 63 patients with metastatic melanoma who did not receive vemurafenib nor immune checkpoint-blocking antibodies. In contrast, in metastatic melanoma patients treated with classical treatment modalities, this CD4+NKG2D+ subset failed to correlate with prognosis. These findings indicate that sorafenib may be used as an "adjuvant" molecule capable of inducing or restoring IL-15Rα/IL-15 in tumors expressing MHCclass I-related chain A/B (MICA/B) and on circulating monocytes of responding patients, hereby contributing to the bioactivity of NKG2D+ Th1 cells.peer-reviewe

Attenuation of Soft-Tissue Sarcomas Resistance to the Cytotoxic Action of TNF-α by Restoring p53 Function

BACKGROUND: Isolated limb perfusion with TNF-α and melphalan is used with remarkable efficiency to treat unresectable limb sarcomas. Here we tested the ability of TNF-α to directly induce apoptosis of sarcoma cells. In addition, we investigated the impact of p53 in the regulation of such effect. METHODOLOGY/PRINCIPAL FINDINGS: We first analysed the ability of TNF-α to induce apoptosis in freshly isolated tumour cells. For this purpose, sarcoma tumours (n = 8) treated ex vivo with TNF-α were processed for TUNEL staining. It revealed substantial endothelial cell apoptosis and levels of tumour cell apoptosis that varied from low to high. In order to investigate the role of p53 in TNF-α-induced cell death, human sarcoma cell lines (n = 9) with different TP53 and MDM2 status were studied for their sensitivity to TNF-α. TP53(Wt) cell lines were sensitive to TNF-α unless MDM2 was over-expressed. However, TP53(Mut) and TP53(Null) cell lines were resistant. TP53 suppression in TP53(Wt) cell lines abrogated TNF-α sensitivity and TP53 overexpression in TP53(Null) cell lines restored it. The use of small molecules that restore p53 activity, such as CP-31398 or Nutlin-3a, in association with TNF-α, potentiated the cell death of respectively TP53(Mut) and TP53(Wt)/MDM2(Ampl). In particular, CP-31398 was able to induce p53 as well as some of its apoptotic target genes in TP53(Mut) cells. In TP53(Wt)/MDM2(Ampl) cells, Nutlin-3a effects were associated with a decrease of TNF-α-induced NF-κB-DNA binding and correlated with a differential regulation of pro- and anti-apoptotic genes such as TP53BP2, GADD45, TGF-β1 and FAIM. CONCLUSION/SIGNIFICANCE: More effective therapeutic approaches are critically needed for the treatment of unresectable limb sarcomas. Our results show that restoring p53 activity in sarcoma cells correlated with increased sensitivity to TNF-α, suggesting that this strategy may be an important determinant of TNF-α-based sarcomas treatment

Genomic expression and single-nucleotide polymorphism profiling discriminates chromophobe renal cell carcinoma and oncocytoma

<p>Abstract</p> <p>Background</p> <p>Chromophobe renal cell carcinoma (chRCC) and renal oncocytoma are two distinct but closely related entities with strong morphologic and genetic similarities. While chRCC is a malignant tumor, oncocytoma is usually regarded as a benign entity. The overlapping characteristics are best explained by a common cellular origin, and the biologic differences between chRCC and oncocytoma are therefore of considerable interest in terms of carcinogenesis, diagnosis and clinical management. Previous studies have been relatively limited in terms of examining the differences between oncocytoma and chromophobe RCC.</p> <p>Methods</p> <p>Gene expression profiling using the Affymetrix HGU133Plus2 platform was applied on chRCC (n = 15) and oncocytoma specimens (n = 15). Supervised analysis was applied to identify a discriminatory gene signature, as well as differentially expressed genes. High throughput single-nucleotide polymorphism (SNP) genotyping was performed on independent samples (n = 14) using Affymetrix GeneChip Mapping 100 K arrays to assess correlation between expression and gene copy number. Immunohistochemical validation was performed in an independent set of tumors.</p> <p>Results</p> <p>A novel 14 probe-set signature was developed to classify the tumors internally with 93% accuracy, and this was successfully validated on an external data-set with 94% accuracy. Pathway analysis highlighted clinically relevant dysregulated pathways of c-erbB2 and mammalian target of rapamycin (mTOR) signaling in chRCC, but no significant differences in p-AKT or extracellular HER2 expression was identified on immunohistochemistry. Loss of chromosome 1p, reflected in both cytogenetic and expression analysis, is common to both entities, implying this may be an early event in histogenesis. Multiple regional areas of cytogenetic alterations and corresponding expression biases differentiating the two entities were identified. Parafibromin, aquaporin 6, and synaptogyrin 3 were novel immunohistochemical markers effectively discriminating the two pathologic entities.</p> <p>Conclusions</p> <p>Gene expression profiles, high-throughput SNP genotyping, and pathway analysis effectively distinguish chRCC from oncocytoma. We have generated a novel transcript predictor that is able to discriminate between the two entities accurately, and which has been validated both in an internal and an independent data-set, implying generalizability. A cytogenetic alteration, loss of chromosome 1p, common to renal oncocytoma and chRCC has been identified, providing the opportunities for identifying novel tumor suppressor genes and we have identified a series of immunohistochemical markers that are clinically useful in discriminating chRCC and oncocytoma.</p

A simplified interventional mapping system (SIMS) for the selection of combinations of targeted treatments in non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is a leading cause of death worldwide. Targeted monotherapies produce high regression rates, albeit for limited patient subgroups, who inevitably succumb. We present a novel strategy for identifying customized combinations of triplets of targeted agents, utilizing a simplified interventional mapping system (SIMS) that merges knowledge about existent drugs and their impact on the hallmarks of cancer. Based on interrogation of matched lung tumor and normal tissue using targeted genomic sequencing, copy number variation, transcriptomics, and miRNA expression, the activation status of 24 interventional nodes was elucidated. An algorithm was developed to create a scoring system that enables ranking of the activated interventional nodes for each patient. Based on the trends of co-activation at interventional points, combinations of drug triplets were defined in order to overcome resistance. This methodology will inform a prospective trial to be conducted by the WIN consortium, aiming to significantly impact survival in metastatic NSCLC and other malignancies

Developpement d'un microdosage de l'adenosine desaminase : application a la mise au point d'un test colorimetrique de cytotoxicite cellulaire et a l'etude de la regulation de l'expression de l'enzyme dans les lymphocytes T cytotoxiques murins ..

SIGLECNRS T 59372 / INIST-CNRS - Institut de l'Information Scientifique et TechniqueFRFranc

Thyroid and Molecular Testing. Advances in Thyroid Molecular Cytopathology

Thyroid nodules are a common finding in the adult population including the fact that more than 50% of individuals, over the age of 60, have thyroid nodules. The majority have been mostly detected with ultrasonography and 10% by palpation. The majority of these nodules are benign, whereas 5–15% of them are malignant. The pre-operative diagnosis of cancer is a critical challenge in order to ensure that each patient can be treated with the best tailored management with a reduction of unnecessary surgery for benign lesions. Fine needle aspiration cytology (FNAC) represents the first and most important diagnostic tool for the evaluation of thyroid lesions. According to the literature, FNAC is able to render a conclusive diagnosis in up to 70–80% of all cases. For the remaining 20–30% of nodules, cytological diagnoses fall into the category of indeterminate lesions mostly due to the lack of specific morphological features. According to the Bethesda system for reporting thyroid cytopathology (TBSRTC), indeterminate lesions can be sub-stratified into three different subcategories including “atypia of undetermined significance/follicular lesion of undetermined significance-AUS/FLUS”; “follicular or Hürthle cell neoplasm/suspicious for follicular or Hürthle cell neoplasm-FN/SFN”; and “suspicious for malignancy-SFM”. Many of these indeterminate lesions undergo repetition or diagnostic lobectomy. Nonetheless, the majority of these cases will have a benign diagnosis due to the fact that the rate of cancer ranges between 6 and 30%. It stands to reason that the application of ancillary technique, mostly molecular testing, emerged as a critical additional tool for those thyroid indeterminate lesions. Since the early 1990s, material collected from cytological samples yields sufficient and adequate cells for the detection of point mutation or gene fusions. Nonetheless, the further availability of new sequencing technologies such as next-generation sequencing (NGS) has led to more comprehensive molecular applications adopted now in clinical use. The current review investigates the multiple advances in the field of molecular testing applied in thyroid cytology