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Purpose: The epithelial–mesenchymal transition (EMT) is emerging as a critical factor for the progression and metastasis of carcinomas, as well as drug resistance. The T-box transcription factor Brachyury has been recently characterized as a driver of EMT in human carcinoma cells. The purpose of this study was to characterize Brachyury as a potential target for lung cancer therapy.Experimental Design: The expression of Brachyury was evaluated by PCR and by immunohistochemistry in human lung tumors and adult normal tissues. Brachyury gene copy number and promoter methylation status were analyzed in tumor tissues with various levels of Brachyury expression. Lung carcinoma cells' susceptibility to T-cell lysis and EGF receptor (EGFR) kinase inhibition were also evaluated relative to the levels of Brachyury.Results: Our results showed Brachyury protein expression in 41% of primary lung carcinomas, including 48% of adenocarcinomas and 25% of squamous cell carcinomas. With the exception of normal testis and some thyroid tissues, the majority of normal tissues evaluated in this study were negative for the expression of Brachyury protein. Brachyury-specific T cells could lyse Brachyury-positive tumors and the level of Brachyury corresponded to resistance of tumor cells to EGFR kinase inhibition.Conclusion: We hypothesize that the elimination of Brachyury-positive tumor cells may be able to prevent and/or diminish tumor dissemination and the establishment of metastases. The ability of Brachyury-specific T-cell lines to lyse Brachyury-positive tumor cells, in vitro, supports the development of Brachyury-based immunotherapeutic approaches for the treatment of lung cancer. Clin Cancer Res; 18(14); 3868–79. ©2012 AACR.

PDF file, 62KB, Analysis of Brachyury gene copy number and promoter methylation status.

PDF file, 55KB, Number and percentage of tissues from adult non-cancer subjects that were positive for Brachyury expression by immunohistochemistry.

PDF file, 113KB, Patient characteristics for samples in human lung cancer cDNA arrays.

Graphical abstract, Highlights • Medication overuse is related to chronicization and medication-overuse headache. • Prediction of medication overuse (MO) is a challenge in the management of migraine. • Machine learning and random optimization could help to estimate MO risk in migraine. • A customized decision support system was devised for migraine clinical management. • This approach may exploit significant patterns in data connoting causality., Machine learning (ML) is largely used to develop automatic predictors in migraine classification but automatic predictors for medication overuse (MO) in migraine are still in their infancy. Thus, to understand the benefits of ML in MO prediction, we explored an automated predictor to estimate MO risk in migraine. To achieve this objective, a study was designed to analyze the performance of a customized ML-based decision support system that combines support vector machines and Random Optimization (RO-MO). We used RO-MO to extract prognostic information from demographic, clinical and biochemical data. Using a dataset of 777 consecutive migraine patients we derived a set of predictors with discriminatory power for MO higher than that observed for baseline SVM. The best four were incorporated into the final RO-MO decision support system and risk evaluation on a five-level stratification was performed. ROC analysis resulted in a c-statistic of 0.83 with a sensitivity and specificity of 0.69 and 0.87, respectively, and an accuracy of 0.87 when MO was predicted by at least three RO-MO models. Logistic regression analysis confirmed that the derived RO-MO system could effectively predict MO with ORs of 5.7 and 21.0 for patients classified as probably (3 predictors positive), or definitely at risk of MO (4 predictors positive), respectively. In conclusion, a combination of ML and RO – taking into consideration clinical/biochemical features, drug exposure and lifestyle – might represent a valuable approach to MO prediction in migraine and holds the potential for improving model precision through weighting the relative importance of attributes.

In clinical practice, inflammatory pain is an important, unresolved health problem, despite the utilization of non-steroidal anti-inflammatory drugs (NSAIDs). In the last decade, different studies have proven that reactive oxygen species (ROS) and reactive nitrogen species (RNS) are involved in the development and maintenance of inflammatory pain and hyperalgesia via the post-translation modification of key proteins, such as manganese superoxide dismutase (MnSOD). It is well-known that inducible cyclooxygenase 2 (COX-2) plays a crucial role at the beginning of the inflammatory response by converting arachidonic acid into proinflammatory prostaglandin PGE2 and then producing other proinflammatory chemokines and cytokines. Here, we investigated the impact of oxidative stress on COX-2 and prostaglandin (PG) pathways in paw exudates, and we studied how this mechanism can be reversed by using antioxidants during hyperalgesia in a well-characterized model of inflammatory pain in rats. Our results reveal that during the inflammatory state, induced by intraplantar administration of carrageenan, the increase of PGE2 levels released in the paw exudates were associated with COX-2 nitration. Moreover, we showed that the inhibition of ROS with Mn (III) tetrakis (4-benzoic acid) porphyrin(MnTBAP) antioxidant prevented COX-2 nitration, restored the PGE2 levels, and blocked the development of thermal hyperalgesia.

Background: Migraine is the most common neurological disorder and the second most disabling human condition, whose pathogenesis is favored by a combination of genetic, epigenetic, and environmental factors. In recent years, several efforts have been made to identify reliable biomarker(s) useful to monitor disease activity and/or ascertain the response to a specific treatment. Objective: To review the current evidence on the potential biological markers associated with migraine. Methods: A structured search of peer-reviewed research literature was performed by searching major publications databases up to December 2017. Results: Several circulating biomarkers have been proposed as diagnostic or therapeutic tools in migraine, mostly related to migraine’s inflammatory pathophysiological aspects. Nonetheless, their detection is still a challenge for the scientific community, reflecting, at least in part, disease complexity and clinical diagnostic limitations. At the present time, calcitonin generelated peptide (CGRP) represents probably the most promising candidate as a diagnostic and/or therapeutic biomarker, as its plasma levels are elevated during migraine attack and decrease during successful treatment. Other molecules (including some neuropeptides, cytokines, adipokines, or vascular activation markers) despite promising, do not possess the sufficient prerequisites to be considered as migraine biomarkers. Conclusion: The characterization of migraine-specific biomarkers would be fundamental in a perspective of precision medicine, enabling risk assessment and tailored treatments. However, speculating on the clinical validity of migraine biomarkers may be premature and controlled clinical trials are presently needed to investigate both the diagnostic and therapeutic value of these biomarkers in migraine.

Background/aim To investigate whether neutrophil-to-lymphocyte ratio (NLR) might represent an additional biological criterion able to identify patients with worse prognosis within the 8th edition TNM prognostic staging system for breast cancer (BC). Patients and methods Pre-treatment NLR was retrospectively analyzed in 475 BC women prospectively followed for a mean time of 3.8 years. The optimal NLR cutoff, identified by ROC analysis, was set at 2. Results Elevated pre-treatment NLR was associated with worse disease-free survival (DFS) (HR=2.28) and overall survival (OS) (HR=3.39). The prognostic value of NLR was mostly evident in stage I BC (HR for DFS=2.89; HR for OS=1.30), in whom NLR significantly stratified patients who developed distant metastasis (HR= 4.62), but not local recurrence. Conclusion NLR might provide important information in risk stratification, especially in stage I BC patients in whom the presence of a high NLR might raise the question as to whether they should be more aggressively managed.

Migraine has been associated with an increased risk for stroke, especially in young women.1 Among the pathophysiologic mechanisms linking migraine and stroke, coagulation abnormalities have been regarded as a logical link.2

Aim: In the present case–control study, we investigated the correlation between the common ACE insertion/deletion (I/D) polymorphism and migraine. Materials & methods: Genotyping of the ACE I/D variant was performed in 502 Caucasian patients with migraine and 323 age-, sex- and race/ethnicity-matched healthy controls. We investigated associations between ACE genetic variants and sociodemographic and/or clinical features of migraineurs. Results: We found a significant association between ACE insertion/insertion (I/I) polymorphism and lower use of pharmacological prophylaxis in migraine patients with aura and in those with chronic migraine. Moreover, ACE I/I polymorphism was significantly more common in migraine patients with aura who had a negative family history of migraine. Conclusion: Our data suggest that although the ACE I/D polymorphism is not a direct risk factor for migraine, the ACE I/I genotype may influence the clinical feature of this disease being associated with reduced use of prophylactic agents in patients with migraine with aura and in those with chronic migraine. Original submitted 5 June 2013; Revision submitted 16 September 2013

The development of Biobanks and recent advances in molecular biology have enhanced the possibility to accelerate translational research studies. The Interinstitutional Multidisciplinary BioBank (BioBIM) is organized in a large healthy donors collection and pathology-based biobanks with the aim to provide a service for development of interdisciplinary studies. A new pathology-based biobank has been organized to specifically collect biospecimen from patients affected by migraine, with the final goal to centralize data, collect blood, plasma, serum, DNA and RNA of patients with this disease. The BioBIM is fully equipped for the automation of sampling/processing, storage and tracking of biospecimens. Standard Operating Procedures have been developed for processing and storage phases as well as archive of clinical data. The availability of biospecimens and clinical data will constitute a resource for various research projects.

Multicenter studies and biobanking projects require blood transportation from the participating center to a central collection or diagnostic laboratory. The impact of time delays between venous blood collection and peripheral blood mononuclear cells (PBMC) isolation prior to RNA extraction may affect the quality and quantity of isolated nucleic acids for genomic applications. Thus, standard operating procedure (SOP) optimization for the treatment of biological samples before RNA extraction is crucial in a biological repository. In order to define SOPs for whole blood preservation prior to RNA extraction, we sought to determine whether different blood storage times (0, 3, 6, 10, 24, and 30 hours) prior to PBMCs isolation and storage at –80°C, could affect the quality and quantity of extracted RNA. After spectrophotometric quantification, the quality and integrity of RNA were assessed by agarose gel electrophoresis, RNA integrity number and real time-PCR (RT-PCR). Across the different time points we did not observe significant differences within the first 24 hours of blood storage at room temperature, while a significant loss in RNA yield and integrity was detected between 24 and 30 hours. We conclude that time delays before PBMCs isolation prior to RNA extraction may have a significant impact on downstream molecular biological applications.

Familial adenomatous polyposis (FAP) is an autosomal-dominant condition mainly due to a mutation of the adenomatous polyposis coli (APC) gene. The present study reports evidence of a technical issue occurring during the mutational analysis of APC exon 4. Genetic conventional direct sequence analysis of a repetitive AT-rich region in the splice acceptor site of APC intron 3 could be misinterpreted as a pathogenetic frameshift result. However, this potential bias may be bypassed adopting a method for random mutagenesis of DNA based on the use of a triphosphate nucleoside analogues mixture. Using this method as a second-level analysis, we also demonstrated the nonpathogenic nature of the variant in the poly A trait in APC exon 4 region (c.423-4delA) that do not result in aberrant splicing of APC exons 3-4; conversely, we did not find a previously reported T deletion/insertion polymorphism.

Vascular endothelial growth factor (VEGF₁₆₅) is stored, transported and released by platelets. Platelet functional abnormalities have been described in patients with hepatocellular carcinoma (HCC). Thus, this study was designed to investigate the behavior of VEGF₁₆₅ with respect to platelet activation in HCC.Plasma and serum VEGF₁₆₅ and plasma sP-selectin levels were analyzed in patients with HCC (n=70) or cirrhosis (n=45) and control subjects (n=70). Given the thrombocytopenia that characterizes both HCC and cirrhotic patients, plasma VEGF₁₆₅ and sP-selectin as well as serum VEGF (plt-VEGF₁₆₅-load) levels were normalized by platelet counts.Median concentrations of plasma VEGF₁₆₅/platelet (p=0.002) and sP-selectin/platelet (p0.0001) were higher in HCC or cirrhotic patients compared to controls. Moreover, sP-selectin/platelet was the only independent variable predictive of plasma VEGF₁₆₅/platelet at multivariate analysis (p0.0001). Conversely, plt-VEGF₁₆₅-load correlated with tumor diameter (p0.05) but not with sP-selectin/platelet and was an independent predictor for 5year overall survival (p=0.012).The results obtained are suggestive for VEGF₁₆₅ release by tumor in HCC. It is plt-VEGF₁₆₅-load, but not plasma VEGF₁₆₅ or serum VEGF₁₆₅ that is an independent predictor for overall survival of HCC patients.

To investigate whether sCD40L dosage might represent a useful tool to explore in vivo platelet function.sCD40L and sP-selectin levels and light transmission aggregometry (LTA) were analyzed in 69 healthy donors. Immunoassays were performed on platelet-depleted citrate plasma samples. The effects of in vitro aspirin treatment on the release of sCD40L were investigated in 15 subjects following platelet stimulation. The effects of a 1-month therapeutic course of low-dose aspirin on sP-selectin and sCD40L levels were also investigated.A significant correlation was observed between sCD40L and sP-selectin (p0.01). In vitro aspirin treatment remarkably decreased sCD40L levels following platelet activation by exogenous agonists. sCD40L directly correlated with LTA (Rho=0.62, p0.0001). In vivo aspirin treatment significantly reduced both sP-selectin and sCD40L levels (both p0.01) in a direct correlation (Rho=0.66, p0.05).Citrated plasma samples reflect sCD40L released from platelets, thus yielding the most valid estimates of in vivo circulating levels of this platelet activation markers.

Mutations in the TP53 gene are the most common genetic alterations in cancer. Accumulation of mutated protein may induce circulating anti-p53 antibodies (anti-p53Ab) in sera of cancer patients. The aim of our work was to evaluate the presence and prognostic value of anti-p53Ab in gastric cancer patients and to investigate whether their presence is related to p53 overexpression in tumor tissue. Anti-p53Ab were analyzed in sera from 111 patients with gastric carcinoma and from 64 healthy donors by ELISA. p53 expression was also quantified by ELISA in biopsies of 54 gastric cancers and 22 healthy gastric mucosas. Significant anti-p53Ab levels were found in 15.3% of patients, whereas none of the 64 donor sera were positive. High levels of p53 expression were detected only in tumor tissue, in 72.2% of cases. A significant correlation was observed between anti-p53Ab and high levels of mutated p53 in tissue (p

The availability of biological samples has assumed a crucial role in the field of biomedical research in order to assess predisposition to complex diseases, identification and validation of new diagnostic biomarkers, drug targets, and improvement of monitoring strategies. To ensure correct collection, storage, and use of biological material, a biobank requires stan- dard operating procedures, not only to guarantee the integrity of the sample, but also for the traceability and uniqueness of the same sample and to avoid potential contamination. Over the past three decades, researchers have increasingly used DNA polymorphisms for biological identification purposes, radically changing forensic investigations. This revolution has led to the birth of new scientific terms, such as "DNA fingerprinting" or "genetic profiling", and a new field of legal medicine known as "forensic genetics". Given the significant need for a biobank to devise a system of identification, authentication, and traceability of biological samples, many researchers are applying forensic genetics-based methodologies to the science of biobanking. This review recapitulates the main DNA polymorphisms presently known and the techniques most widely used for biological identification purposes, from standard methods based on short tandem repeats, to use of single nucleotide polymorphisms, through to the more recent insertion/ deletion polymorphisms. The experience of several research groups who have applied classical or innovative methods for identification of biological samples in the context of a biorepository is also discussed. Finally, some technical issues are suggested that could facilitate decisions about the most appropriate markers and DNA fingerprint methodology for evaluation of identity and traceability in each disease-based biobank.

Aim: This study was designed to assess the prognostic significance of the combined measurement of vascular endothelial growth factor (VEGF) and carcinoembryonic antigen (CEA) tissue content with respect to relapse-free and overall survival of patients with colorectal cancer (CRC). Methods: Quantitative evaluation of VEGF and CEA content was performed on protein extracts obtained from tissue biopsies from 69 CRC patients and 15 healthy donors. Results: VEGF significantly correlated with CEA content of either tumor tissues (rho = 0.55, p < 0.0001) or corresponding normal mucosa (rho = 0.34, p < 0.005). General regression analyses demonstrated that CEA was an independent predictor of VEGF tissue content either in CRC biopsies (regression coefficient = 0.57, p < 0.0001) or normal mucosa (regression coefficient = 0.25, p < 0.05). Cox proportional hazards survival analysis showed that tumor tissue content of both VEGF and CEA had an independent prognostic value in predicting both relapse-free (hazards ratio = 5.98, p = 0.002) and overall (hazards ratio = 4.73, p = 0.007) survival, irrespective of Dukes’ stage. Kaplan-Meier analysis demonstrated that an elevated tumor content of both CEA and VEGF had a negative prognostic value in respect to either relapse-free (log-rank test: 10.4, p = 0.001) or overall survival (log-rank test: 7.33, p = 0.007). Conclusion: Tumor tissue VEGF and CEA content determination might add useful prognostic information in the management of patients with CRC.

Objectives: To analyse the behaviour of pre-surgical serum levels of soluble (s)E-selectin and vascular cell adhesion molecule (sVCAM) in patients with colorectal cancer, and to evaluate their possible correlation with carcinoembryonic antigen (CEA), pro-inflammatory cytokines and clinicopathological features with respect to their prognostic value in predicting metastatic disease. Methods: Pre-surgical serum levels of sE-selectin, sVCAM, interleukin-6 (IL-6), IL-1β, tumour necrosis factor-α (TNF-α) and CEA were measured in 194 patients with colorectal adenocarcinoma, 40 patients with benign colorectal diseases and 59 healthy subjects. Results: sE-selectin, sVCAM, TNF-α and IL-6 levels were significantly higher in patients with colorectal cancer compared to either healthy subjects or patients with benign disease. Positive rates of sE-selectin, sVCAM and TNF-α levels were significantly associated with Dukes’ stage D colorectal cancer, and all three variables were independently associated to the presence of distant metastases. Positive sE-selectin, sVCAM and TNF-α levels were significantly associated to CEA. TNF-α and CEA levels were independently related to the presence of positive levels of sE-selectin and/or sVCAM. Conclusions: Our findings suggest that the host inflammatory response to cancer cells, and/or their released products (i.e. CEA), might be responsible (via cytokine release) for the elevation in circulating adhesion molecules in patients with colorectal cancer.

Increased expression of selectins has been found on endothelial cells of venules and capillaries in the tumor stroma of non-small cell lung cancer, suggesting their functional role in the process of chemotaxis for tumor cells. The present study was aimed at analyzing the role of both soluble (s)P-selectin and sE-selectin levels in association with clinico-pathological variables in 116 patients with lung cancer, 38 patients with benign diseases and 59 healthy donors. The results obtained showed that sP-selectin and sE-selectin levels were higher in patients with lung cancer compared to normal donors (p

AIM To investigate the possible predictive role of routinely used glycemic parameters for a first venous thromboembolism (VTE) episode in gastrointestinal (GI) cancer ambulatory patients - with or without clinically diagnosed type 2 diabetes (T2D) or obesity - treated with chemotherapy. METHODS Pre-treatment fasting blood glucose, insulin, glycated hemoglobin (HbA1c) and homeostasis model of risk assessment (HOMA) were retrospectively evaluated in a cohort study of 342 GI cancer patients. Surgery was performed in 142 (42%) patients with primary cancer, 30 (21%) and 112 (79%) of whom received neoadjuvant and adjuvant therapies, respectively. First-line chemotherapy was administered in 200 (58%) patients with metastatic disease. The study outcome was defined as the occurrence of a first symptomatic or asymptomatic VTE episode during active treatment. RESULTS Impaired glucose tolerance (IGT) or T2D were diagnosed in 30% of GI cancer patients, while overweight/obesity had an incidence of 41%. VTE occurred in 9.4% of patients (7% of non-diabetic non-obese), especially in those with a high ECOG score (P = 0.025). No significant association was found between VTE incidence and T2D, obesity, different tumor types, metastatic disease, Khorana class of risk, or different anti-cancer drugs, although VTE rates were substantially higher in patients receiving bevacizumab (17% vs 8%, P = 0.044). Conversely, all glucose metabolic indexes were associated with increased VTE risk at ROC analysis. Multivariate Cox proportional analyses confirmed that HOMA index (HR = 4.13, 95%CI: 1.63-10.5) or fasting blood glucose (HR = 3.56, 95%CI: 1.51-8.39) were independent predictors of VTE occurrence during chemotherapy. CONCLUSION The results here reported demonstrate that evaluating glucose metabolic asset may allow for VTE risk stratification in GI cancer, helping to identify chemotherapy-treated patients who might benefit from thromboprophylaxis. Further multicenter prospective studies involving a larger number of patients are presently needed.

The epithelial–mesenchymal transition (EMT) is a normal developmental process that allows the conversion of epithelial, polarized, and stationary cells into highly motile and invasive mesenchymal cells, a phenomenon required for the normal formation of the embryo (1,2). Recent investigations have demonstrated that a similar phenotypic switch can aberrantly take place during tumor progression, allowing epithelial cancer cells to lose cell polarity, epithelial markers, and cell-to-cell contacts, while simultaneously acquiring mesenchymal-associated markers, cell motility, and invasiveness (3,4). As a result, carcinoma cells undergoing EMT are enabled to initiate the cascade of events leading to the establishment of distant metastases (5). In breast cancer, the phenomenon of EMT has been shown to take place in highly aggressive tumors with a basal-like phenotype, which frequently show overexpression of mesenchymal proteins, including vimentin and smooth-muscle-actin, and cadherin switching (reduced E-cadherin and overexpression of N-cadherin) (6,7). In breast cancer patients undergoing neoadjuvant therapy, moreover, it has been shown that recurring tumors after treatment with standard anthracycline-taxane chemotherapy had an increased proportion of cells expressing EMT-associated genes, thus highlighting the importance of EMT in therapeutic resistance (8). We have identified brachyury, an embryonic transcription factor of the T-box family (9,10), as a driver of EMT in human carcinomas (11,12). High levels of brachyury expression in epithelial tumor cells have been shown to drive the acquisition of mesenchymal features in vitro, including motility and invasiveness, and to favor metastatic dissemination of tumor cells in vivo (11,12). In a recent study conducted with a monoclonal murine antibody, expression of brachyury protein was found in approximately 40% of non–small cell lung carcinomas and not in a range of human adult normal tissues, with the exception of low levels observed in testis and selected thyroid samples (13,14). We have been able to demonstrate the generation of brachyury-specific T cells capable of lysing human lung carcinoma cells (13). A phase I study is ongoing using a brachyury-based vaccine in patients with metastatic tumors (15). In this study we characterize the expression of brachyury in a range of malignant and benign breast tissues both at the mRNA and protein levels and present data on the potential role of brachyury in the biology of breast cancer. We also demonstrate the ability of brachyury-specific T cells to lyse human breast carcinoma cells, providing the rationale for the use of a brachyury-specific vaccine as a monotherapy or in combination therapy approaches for breast carcinoma.

The efficacy of CEA and CA15-3 tumor markers in monitoring breast cancer was evaluated in 1365 patients with either benign (n = 534) or malignant (n = 831) breast diseases. Thirty-nine breast cancer patients were monitored before and after neoadjuvant chemotherapy. Three hundred forty-nine patients were monitored during post-surgical follow-up for either a minimum of 5 years or until time of recurrence. Twenty-one patients with metastases were also monitored during chemotherapy. Elevated CA 15-3 and TPS levels were found in 28.6% and 30.0% of patients. CA 15-3 and TPS sensitivities rose to 71.9% and 66.3% in metastatic patients, respectively. The addition of TPS to CA 15-3 increased the sensitivity up to 44.4% in the overall population, and to 87.6% in patients with metastases. During post-surgical follow-up CA 15-3 was elevated in 65.7% and TPS in 61.3% of patients with recurrence. The combination of TPS and CA 15-3 increased the overall sensitivity by 12.7%. Longitudinal monitoring of metastatic patients undergoing chemotherapy demonstrated that, when positive, both CA 15-3 and TPS paralleled response to treatment. TPS monitoring may provide additional value when used in combination with CA15-3 during post-surgical follow-up of breast cancer patients.

Several lines of investigation have revealed the apparent interplay between the immune system of the host and many conventional, "standard-of-care" anticancer therapies, including chemotherapy and small molecule targeted therapeutics. In particular, preclinical and clinical studies have demonstrated the important role of regulatory T cells (Tregs) in inhibiting immune responses elicited by immunotherapeutic regimens such as those based on anticancer vaccines or checkpoint inhibitors. However, how the number and immunosuppressive function of Tregs change in cancer patients undergoing treatment with non-immune anticancer therapies remains to be precisely elucidated. To determine whether immunostimulatory therapies can be employed successfully in combination with conventional anticancer regimens, we have investigated both the number and function of Tregs obtained from the peripheral blood of carcinoma patients before the initiation and during the course of chemotherapeutic and targeted agent regimens. Our studies show that the treatment of breast cancer patients with tamoxifen plus leuprolide, a gonadotropin releasing hormone agonist, has minimal effects on Tregs, while sunitinib appears to exert differential effects on Tregs among patients with metastatic renal carcinoma. However, the administration of docetaxel to patients with metastatic prostate or breast cancer, as well as that of cisplatin plus vinorelbine to non-small cell lung cancer patients, appears to significantly increase the ratio between effector T cells and Tregs and to reduce the immunosuppressive activity of the latter in the majority of patients. These studies provide the rationale for the selective use of active immunotherapy regimens in combination with specific standard-of-care therapies to achieve the most beneficial clinical outcome among carcinoma patients.

Standard operating procedures (SOPs) optimization for nucleic acid extraction from stored samples is of crucial importance in a biological repository, considering the large number of collected samples and their future downstream molecular and biological applications. However, the validity of molecular studies using stored specimens depends not only on the integrity of the biological samples, but also on the procedures that ensure the traceability of the same sample, certifying its uniqueness, and ensuring the identification of potential sample contaminations. With this aim, we have developed a rapid, reliable, low-cost, and simple DNA fingerprinting tool for a routine use in quality control of biorepositories samples. The method consists of a double ALU insertion/deletion genotyping panel suitable for uniqueness, identification of sample contaminations, and gender validation. Preliminary data suggest that this easy-to-use DNA fingerprinting protocol could routinely provide assurances of DNA identity and quality in a biorepository setting.

AIM To investigate the clinical significance of routinely used glycemic parameters in a cohort of colorectal cancer (CRC) patients. METHODS Pre-treatment fasting blood glucose, insulin, HbA1c and homeostasis model of risk assessment (HOMA-IR) were retrospectively evaluated in a case-control study of 224 CRC and 112 control subjects matched for sex, obesity and diabetes frequency and blood lipid profile. Furthermore, the prognostic value of routinely used glycemic parameters towards progression-free (PFS) and overall survival (OS) was prospectively evaluated. RESULTS Fasting blood glucose, insulin, HOMA-IR and HbA1c (all P < 0.0001) levels were higher in non-diabetic CRC patients compared with obesity-matched controls. All parameters were associated with increased CRC risk at ROC analysis, but no relationship with clinical-pathological variables or survival outcomes was observed for glycemia, insulinemia or HOMA-IR. Conversely, advanced CRC stage (P = 0.018) was an independent predictor of increased HbA1c levels, which were also higher in patients who had disease progression compared with those who did not (P = 0.05). Elevated HbA1c levels showed a negative prognostic value both in terms of PFS (HR = 1.24) and OS (HR = 1.36) after adjustment for major confounders, which was further confirmed in a subgroup analysis performed after exclusion of diabetic patients. CONCLUSION HbA1c might have a negative prognostic value in CRC, thus suggesting that glycemic metabolic markers should be carefully monitored in these patients, independently of overt diabetes.

Biobanks provide stored material to basic, translational, and epidemiological research and this material should be transferred without institute-dependent intrinsic bias. The ISBER Biospecimen Science Working Group has released a “Standard PREanalytical Code” (SPREC), which is a proposal for a standard coding of the preanalytical options that have been adopted in order to track and make explicit the preanalytical variations in the collection, preparation, and storage of specimens. In this paper we address 2 issues arising in any biobank or biolaboratory aiming at adopting SPREC: (i) reducing the burden required to adopt this standard coding, and (ii) maximize the immediate benefits of this adoption by providing a free, dedicated software tool. We propose SPRECware, a vision encompassing tools and solutions for the best exploitation of SPREC based on information technology ( www.sprecware.org ). As a first step, we make available SPRECbase, a software tool useful for generating, storing, managing, and exchanging SPREC-related information associated to specimens. Adopting SPREC is useful both for internal purposes (such as finding the samples having some given preanalytical features), and for exchanging the preanalytical information associated to biological samples between Laboratory Information Systems. In case of a common adoption of this coding, it would be easy to find out whether and where, among the participating Biological Resource Centers, the specimens for a given study are available in order to carry out a planned experiment.

Purpose: The epithelial–mesenchymal transition (EMT) is emerging as a critical factor for the progression and metastasis of carcinomas, as well as drug resistance. The T-box transcription factor Brachyury has been recently characterized as a driver of EMT in human carcinoma cells. The purpose of this study was to characterize Brachyury as a potential target for lung cancer therapy. Experimental Design: The expression of Brachyury was evaluated by PCR and by immunohistochemistry in human lung tumors and adult normal tissues. Brachyury gene copy number and promoter methylation status were analyzed in tumor tissues with various levels of Brachyury expression. Lung carcinoma cells' susceptibility to T-cell lysis and EGF receptor (EGFR) kinase inhibition were also evaluated relative to the levels of Brachyury. Results: Our results showed Brachyury protein expression in 41% of primary lung carcinomas, including 48% of adenocarcinomas and 25% of squamous cell carcinomas. With the exception of normal testis and some thyroid tissues, the majority of normal tissues evaluated in this study were negative for the expression of Brachyury protein. Brachyury-specific T cells could lyse Brachyury-positive tumors and the level of Brachyury corresponded to resistance of tumor cells to EGFR kinase inhibition. Conclusion: We hypothesize that the elimination of Brachyury-positive tumor cells may be able to prevent and/or diminish tumor dissemination and the establishment of metastases. The ability of Brachyury-specific T-cell lines to lyse Brachyury-positive tumor cells, in vitro, supports the development of Brachyury-based immunotherapeutic approaches for the treatment of lung cancer. Clin Cancer Res; 18(14); 3868–79. ©2012 AACR.

PURPOSE: The aim of this study was to evaluate the possibility of identifying biologically aggressive subgroups of patients, combining several biologic parameters such as the tumor and normal mucosa values from the ploidy, the S-phase cell percentage, and tumor-associated glycoprotein-72 (TAG-72) expression. METHODS: One hundred five colorectal cancer patients were studied to analyze the possible prognostic role of ploidy and cell kinetics in multiple fresh specimens from the tumor and normal mucosa. Since the presence of TAG-72 in the colonic mucosa has been correlated to neoplastic transformation, the correlations between these parameters and the quantitative tissue expression of the TAG-72 were analyzed in a subgroup of 53 cases. RESULTS: A significantly lower five-year disease-free survival rate (21.4 percent) was observed in patients with multiploid tumors, when compared with that observed in patients with diploid or single aneuploid tumors (67.5 percent) (P=0.03). The quantitative tissue TAG-72 expression contributed in identifying a particular patient subgroup (20 percent), characterized by S-phase percentage and TAG-72 values from the normal mucosa that were unexpectedly higher than 12.1 percent and 7.5 U/mg of proteins, respectively. In particular, when the 25 Dukes B patients were analyzed, similar results were observed. In fact, 14 (56 percent) cases showed high tumor cell proliferation and, surprisingly, a high tissue TAG-72 content in the normal mucosa was found in 4 (28.6 percent) of these patients. CONCLUSIONS: Other than multiploidy, the biologic aggressiveness of colorectal cancer might be successfully assessed introducing the evaluation of new biologic parameters, such as the TAG-72 content and S-phase percentage values of the normal mucosa, suggesting the possibility of further stratifying this patient population.

Aims and Background Sentinel lymph node dissection (SLND) has recently been evaluated as a new staging technique for early breast cancer. To minimize the extent of surgery, the feasibility of eradicating primary breast lesions and the relative sentinel lymph nodes (SLN) under regional anesthesia was evaluated in this study. Methods and Study Design A selected population of 76 patients with suspected operable breast cancer and no clinically palpable lymph nodes was enrolled in the study. Intra- and perilesional administration of a radiotracer was performed. Lymphoscintigraphy was carried out to confirm the drainage pathway and locate the SLN. The following day, after inducing a nervous block induction of the ipsilateral intercostal nerves, we performed the surgical procedure with the help of a hand-held gamma-detecting probe. In case the primary lesion was diagnosed as invasive carcinoma by frozen section, the SLN and the remaining axillary lymph nodes (non-SLNs) were removed. The status of SLN and non-SLNs was compared. Results The primary breast lesion was located and excised in all cases (identification rate: 100%). Lymphoscintigraphy positively identified SLNs in 40/45 (89%) patients; in five patients no lymphatic drainage was detected. In 38 cases an average of 1.5 SLNs and 14 non-SLNs per patient were removed and pathologically analyzed; the remaining two patients showed SLNs in the internal mammary chain, which were not excised. Twenty-nine percent of the patients showed metastatic disease in the lymph nodes examined. Of all patients with affected nodes, 55% had cancer cells only in the SLN. No false negatives (skip metastases) were found. No immediate or long-term anesthesia-related complications (eg pleural lesions, intravascular injection) were observed. Conclusions Our data confirm the feasibility of single radiotracer administration for both occult lesion and SLN localization as well as the usefulness of SLND in staging early breast cancer. Regional anesthesia resulted in easy management and good patient compliance. This time-saving procedure allowed the completion of the whole surgical plan, reducing the recovery time without modifying the effectiveness of surgery.

Preanalytical variables, including the anticoagulants and stabilizing agents, time, storage temperature, and methods of DNA extraction applied to blood samples, may affect quality and quantity of isolated nucleic acids for future genomic applications. Considering the large number of collected samples, standard operating procedure optimization for whole blood preservation before DNA extraction is a crucial step in a biological repository. Moreover, the future application of the biological material may not be known subsequent to its extraction. To define standard operating procedures for whole blood preservation before DNA extraction, we aimed to determine whether different combinations of anticoagulants, blood storage temperatures, and time intervals before storage at -80°C might have an impact on quality and quantity of subsequent extracted DNA. After spectrophotometer quantification, the quality and integrity of DNA were assessed by agarose gel electrophoresis, polymerase chain reaction, and real-time polymerase chain reaction methods. We observed that decrease in DNA recovery during blood storage time was more pronounced at room temperature than at 4°C. Based on our experience, we recommend as anticoagulants of choice sodium citrate and ethylenediaminetetraacetate, whereas sodium citrate theophylline adenosine dipyridamole could represent an alternative choice, excluding a priori lithium heparin and Fluoride-Oxalate. Based on the overall evaluation criteria, we conclude that the procedures necessary to preserve the whole blood before the DNA extraction may have a significant impact on downstream molecular biological applications.

Background Biospecimen quality is crucial for clinical and translational research and its loss is one of the main obstacles to experimental activities. Beside the quality of samples, preanalytical variations render the results derived from specimens of different biobanks or even within the same biobank incomparable. Specimens collected along the years should be managed with a heterogeneous life cycle. Hence, we propose to collect detailed data concerning the whole life cycle of stored samples employing radio-frequency identification (RFID) technology. Methods We describe the processing chain of blood biosamples that is operative at the biobank of IRCSS San Raffaele, Rome, Italy (BioBIM). We focus on the problem of tracing the stages following automated preanalytical processing: we collected the time stamps of all events that could affect the biological quality of the specimens by means of RFID tags and readers. Results We developed a pilot study on a fragment of the life cycle, namely the storage between the end of the preanalytics and the beginning of the analytics, which is usually not traced by automated tools because it typically includes manual handling. By adopting RFID devices we identified the possible critical time delays. At 1, 3 and 6 months RFID-tagged specimens cryopreserved at -80°C were successfully read. Conclusions We were able to record detailed information about the storage phases and a fully documented specimen life cycle. This will allow us to promote and tune up the best practices in biobanking because i) it will be possible to classify sample features with a sharper resolution, which allows future utilization of stored material; ii) cost-effective policies can be adopted in processing, storing and selecting specimens; iii) after using each aliquot, we can study the life cycle of the specimen with a possible feedback on the procedures.

BACKGROUND: This study analyzed the possible prognostic value of presurgical serum soluble (s)E-selectin levels and/or carcinoembryonic antigen (CEA) mRNA positivity in predicting the disease-free survival of colorectal cancer (CRC) patients. METHODS: CEA mRNA (obtained from blood-borne cells by reverse transcriptase-polymerase chain reaction [RT-PCR]), tumor necrosis factor-α (TNF-α), and sE-selectin levels were analyzed in blood samples obtained from 78 patients with primary (n = 62) or recurrent (n = 16) CRC, 40 patients with benign colorectal (CR) diseases, and 78 controls. RESULTS: CEA mRNA positivity by RT-PCR was significantly associated with advanced stage (P < .05). Median baseline sE-selectin levels were higher in patients with CRC (43 ng/mL) compared with controls (36 ng/mL) or patients with benign CR diseases (31 ng/mL, P < .001). These were significantly associated with CEA mRNA positivity by RT-PCR (P < .05). Multivariate analysis by forward stepping showed that elevated TNF-α (P = .001) and CEA mRNA positivity by RT-PCR (P = .0001) were independent predictors of elevated baseline sE-selectin levels. Positive presurgical sE-selectin levels were associated with an increased recurrence rate compared with patients with low levels of this molecule (P < .001). Positivity for both CEA mRNA and sE-selectin had a negative prognostic impact, with a 5-year recurrence-free survival rate of 51% compared with 95% of patients with negative parameters (P < .05). CONCLUSIONS: Detection of presurgical serum sE-selectin levels and CEA mRNA-positive blood-borne cells in CRC patients might provide useful prognostic information in terms of recurrence-free survival, either alone or in combination, and may help in the choice of more aggressive treatment and/or more strict follow-up procedures in high-risk patients. Cancer 2010. © 2010 American Cancer Society.

Adipokines may significantly influence the growth and proliferation of tumor stroma and malignant cells within. Reduced adiponectin and increased leptin serum levels were found in colorectal cancer (CRC) patients. Recently, it has been demonstrated that tumor necrosis factor-alpha (TNF-alpha) is able to induce dose-dependent changes in serum adipokine levels. Thus, aims of this study were to evaluate the possible associations between adipokines, TNF-alpha and clinicopathological variables of CRC patients and to analyze their possible prognostic value in predicting relapse-free and overall survival.Baseline leptin, adiponectin and TNF-alpha levels were analyzed in 90 patients with histologically diagnosed primary or newly diagnosed metastatic CRC treated at 'Tor Vergata' Clinical Center and followed up for a median period of 3 years.Serum leptin levels were higher in CRC patients than in controls (p0.0001). Conversely, serum adiponectin levels were lower in CRC patients than in controls (p0.0001). Leptin inversely correlated with adiponectin (p0.005). The leptin/adiponectin (L/A) ratio was eight-fold greater in CRC compared to controls (p0.0001). Kaplan-Meier analysis of relapse-free and overall survival time showed that the L/A ratio was an independent predictor for adverse outcome in CRC.Serum adipokine levels might have a role in the biology of CRC and the combined measurement of leptin and adiponectin levels might provide useful prognostic information in the management of patients with CRC.

Objectives: A longitudinal study was designed to quantify tumor tissue content of vascular endothelial growth factor (VEGF) in patients with colorectal cancer (CRC) and to evaluate its prognostic value in respect to the relapse-free and overall survivals. Methods: Sixty-nine patients with CRC were followed from the time of diagnosis of primary tumor for at least 3 years after surgery. Quantitative evaluation of VEGF content in tissue was performed on whole protein extracts obtained from biopsies of histologically confirmed neoplastic tissues and corresponding mucosa, histologically confirmed as ‘normal’. Results: VEGF levels were higher in CRC tissues, median 141 pg/mg of protein (interquartile range 70–375), compared with corresponding normal mucosa, median 45 pg/mg of protein (interquartile range 22–78; p < 0.0001), and were associated with the stage of disease (p = 0.035) by multivariate analysis. Tumor VEGF content was higher in relapsing patients compared with those who remained disease free (p < 0.001) and was independently associated with relapse-free survival (p = 0.044). Cox’s proportional hazard survival analysis demonstrated that VEGF (p = 0.035) had an independent prognostic value in respect to overall survival. Conclusions: Elevated tumor VEGF content may discriminate between early and late stages of CRC and may be used as an independent prognostic parameter in the management of these patients.

Purpose: Tumor-induced platelet activation may cause the release of various cytokines, including CD40 ligand (CD40L). Activation of the CD40/CD40L pathway in human tumors may result in thrombin generation, which is known to be involved in angiogenesis. Thus, we investigated whether soluble (s)CD40L levels are increased in patients with lung cancer as a result of platelet and/or coagulation activation. Experimental Design: Citrated plasma samples were obtained from 120 patients with different stages and histotypes of lung cancer and 60 age- and sex-matched control subjects. sCD40L, sP-selectin (marker of platelet activation), prothrombin fragment 1 + 2, and thrombin-antithrombin III complex levels (both markers of coagulative activation) were measured in all samples. Results: Patients with lung cancer had median sCD40L levels higher than in control subjects (0.46 versus 0.13 ng/ml; P < 0.0001), although correlation with the stage of disease was not evident. Nonetheless, sCD40L levels were significantly higher in squamous cancer compared with adenocarcinoma (0.75 versus 0.27 ng/ml; P < 0.05). Moreover, median sCD40L levels were higher in stage IV compared with nonmetastatic squamous lung cancer (1.02 versus 0.61 ng/ml; P < 0.05). sCD40L levels significantly correlated with sP-selectin (P < 0.001), prothrombin fragment 1 + 2 (P < 0.001), or thrombin-antithrombin III complex (P < 0.05) in squamous lung cancer, but only sP-selectin (P = 0.011) was independently related to sCD40L. Conclusions: These findings indicate that elevated sCD40L levels can be preferentially found in patients with advanced squamous cancer and provide evidence that increased levels of this cytokine are associated to the occurrence of in vivo platelet activation.

Measurement of soluble (s) P-selectin levels has been proposed as a diagnostic tool for monitoring the clinical course of human neoplasms. Thus, our study was aimed at analyzing the role of sP-selectin in association with clinicopathological variables in 181 patients with primary (n =149) or metastatic (n = 32) colorectal cancer (CRC), 34 patients with benign diseases and 181 control subjects. The results obtained showed that sP-selectin levels were higher in patients with CRC compared either to patients with benign disease (p = 0.006) or controls (p = 0.003). No differences were observed between the latter and patients with benign diseases. Increased median sP-selectin levels were significantly associated with the presence of distant metastasis (68.2 ng/ml vs. 48.6 ng/ml, p = 0.002). Of interest, carcinoembryonic antigen (CEA) levels were independently associated to sP-selectin (regression coefficient = 0.28, p < 0.002). Cox's proportional hazards survival analysis of primary CRC patients demonstrated that beside the stage of disease sP-selectin levels had an independent prognostic role in predicting recurrent disease (HR = 2.22, p = 0.019) and mortality from CRC (HR = 3.44, p= 0.017). These results suggest that measurement of plasma sP-selectin might represent a prognostic indicator in the management of patients with CRC.

The present study was designed to investigate whether a correlation exists between IL-6, TNF-alpha and coagulation (Thrombin-antithrombin, TATc) or fibrinolysis (D-dimer) activation in non-small cell lung cancer (NSCLC) patients. One hundred thirty patients with NSCLC (n=65, 53 males, mean age 65 +/- 8, adenocarcinoma n=32, squamous cancer n=33) or chronic obstructive pulmonary disease (COPD) (n=65, 51 males, mean age 67 +/- 9) were studied. As control group 65 healthy donors (51 males, mean age 61 +/- 14) were also evaluated. The results obtained showed that median D-dimer levels were higher in NSCLC patients (3.0 microg/ml) compared either to COPD patients (1.1 microg/ml, P0.05) or controls (0.3 microg/ml, P0.0001). Positive TNF-alpha levels (10 pg/ml) were found in 26% of NSCLC compared to 3% of COPD (P0.002) and 5% of controls (P0.0005). On the other hand, positive (8.5 pg/ml) IL-6 levels were found in 53% of NSCLC and 21% of COPD patients, compared to 5% of control subjects (P0.001). Median TATc levels were elevated in either NSCLC (6.9 microg/l) or COPD (5.7 microg/l) patients compared to controls (1.8 microg/l, P0.0001). Elevated D-dimer levels were significantly associated to positive TNF-alpha levels in patients without distant metastasis (F=4.3, P0.05). Moreover, TNF-alpha levels (P0.01) were independently related to the presence of positive D-dimer levels in patients with non-metastatic NSCLC. These results suggest that increased levels of TNF-alpha might be responsible for an activation of fibrinolysis in patients with NSCLC.

Hyperthermic isolated limb perfusion (ILP) with tumor necrosis factor-a (TNFalpha) and cytotoxic drugs is currently used for treatment of melanoma and sarcoma of the limbs. Tumor necrosis factor-alpha is involved in the systemic inflammatory response syndrome as a result of activation of inflammatory cells and production of bioactive substances. The goal of this study was to determine the circulating levels of proinflammatory cytokines and soluble adhesion molecules in 19 patients with limb melanoma or sarcoma undergoing ILP with (n = 9) or without TNFalpha (n = 10). The results obtained demonstrated that ILP with TNFalpha was responsible for a leakage of TNFalpha in the systemic circulation, followed by a rise in interleukin (IL)-6 and IL-8 levels within I h. Elevated soluble (s)P-selectin levels were found 1-3 h after ILP. Plasma sE-selectin peaked 6-9 h after ILP, and soluble vascular cell adhesion molecule (sVCAM) levels reached a maximum after 24 h. Significant correlations were observed among these variables, confirming the interdependence of all changes observed. On the other hand, ILP with cytotoxic drugs alone induced only a modest release of TNFalpha, which was not followed by an immediate rise in IL-6 and IL-8. Four of the 9 patients undergoing ILP with TNF had severe systemic toxicity. No association was found between systemic TNF levels and the clinical outcome, whereas elevated TNF perfusion levels as well as systemic IL-6 and IL-8 levels were constantly elevated in patients with severe toxicity. These results are suggestive of an important role of TNFalpha levels in the perfusion system (more than leakage of perfusate) in causing postoperative toxicity, although other ILP-related factors should not be excluded.

Tissues and sera from 110 patients diagnosed with colorectal primary carcinoma, 20 patients with benign colorectal diseases and 31 healthy donors were subjected to quantitative CEA analysis. Multiple samples from tumor lesions and autologous histologically normal mucosa (10 cm from the tumor) were obtained at the time of surgery (cancer patients) or endoscopy (benign patients and healthy volunteers). CEA content was measured in protein extracts obtained from these tissues using a quantitative RIA method. A limit of normality for CEA content was established as 300 ng/mg of protein. When this was taken as cut-off, 104 of 110 (94.5%) tumor lesions and 51 of 110 (46.4%) autologous histologically normal colonic mucosa from cancer patients had elevated CEA levels. No correlation with stage of disease was found, while a correlation was observed with degree of tumor differentiation. A statistically significant difference between CEA content in tumor lesions and in histologically normal mucosa from cancer patients was observed (p = -0.001). Moreover, CEA content was statistically higher in the normal mucosa from cancer patients than in that from healthy donors (p = 0.005). CEA content in tissue specimens from benign lesions differed significantly from that in tissue from healthy donors (p = 0.005) and in carcinoma lesions (p < 0.001). The highest CEA content was observed in benign lesions with severe dysplasia. No statistical correlation between CEA content in carcinoma tissues and serum CEA levels (r = 0.195, p = .13) was found. Therefore, in considering diagnosis or therapy with anti-CEA MAbs for colorectal-carcinoma patients, or potential therapies with anti-CEA recombinant vaccines, serum CEA levels should not be taken as indicating CEA expression in tumor lesions.

Among the new tumor markers that have been recently proposed, CA 72-4 is of particular interest, not only for its capabilities in diagnosing and monitoring certain neoplastic diseases, but also for its excellent specificity. Several studies focused on the potential clinical usefulness of CA 72-4 in gastrointestinal (GI) and gynecological cancer, showing a sensitivity of approximately 40% in colorectal and gastric cancer and 50% in ovarian cancer, with an overall specificity of more than 95%. Longitudinal evaluations of patients with either GI or gynecological malignant diseases demonstrated that significant elevations of CA 72-4 serum levels may be predictive of recurrent disease. Moreover, the combination of CA 72-4 with other known serum markers, such as CEA and CA 19-9 for GI cancer or CA 125 for ovarian cancer, indicated that an increase in the sensitivity can be achieved without substantial changes in the overall specificity, improving the possibility of monitoring these patients. In conclusion, these results provide a strong argument for the use of CA 72-4 in the management of these neoplastic diseases.

The possible correlation(s) between platelet proaggregating activity, and sialic acid content and ganglioside expression of six human colorectal tumour cell lines (CBS, GEO, HT-29, WiDr, MIP and DLD-1) was evaluated. The three cell lines (HT-29, WiDr and DLD-1) capable of inducing remarkable in vitro platelet aggregation, had significantly higher amounts of lipid-bound sialic acid than those cell lines characterised by a lower platelet proaggregating activity (GEO, CBS and MIP). High performance thin-layer chromatography demonstrated the presence of one band comigrating with GM3 in all cell lines, while GD1a and GT1b comigrating gangliosides were present only in HT-29, WiDr and DLD-1 cells. Finally, an increased platelet pro-aggregating activity of GEO and CBS cell lines was observed after the incorporation of exogenous gangliosides. The present data support the hypothesis that lipid-bound sialic acid may be involved in platelet-tumour cell interactions.

Objective: This study was performed to evaluate the clinical values of tumor-associated glycoprotein-72 serum levels alone or in combination with CA 125 in the diagnosis and monitoring of patients with ovarian cancer. Study Design: Serum samples from 293 patients, 142 with primary carcinoma and 151 with benign diseases of the genital tract, were evaluated for the presence of CA 125, tumor-associated glycoprotein-72, and carcinoembryonic antigen. All patients underwent surgery for the primary tumor, and stage was defined according to the classification of International Federation of Gynecology and Obstetrics. Results: When the measurement of serum tumor-associated glycoprotein-72 is combined with that of CA 125, the sensitivity for the detection of primary ovarian cancer increased from 60% to 73%, with no significant change in specificity, and resulted in a more accurate clinical assessment for detection of residual disease before the second-look procedure. In fact, when both markers were positive, 100% specificity was achieved; conversely, when both markers were negative, no residual disease was found. Conclusion: These findings suggest that tumor-associated glycoprotein-72 may be considered as a supplementary serum marker for CA 125, providing further clinical information for the diagnosis of primary and recurrent ovarian cancer.

Tumor-associated glycoprotein-72 has been recently suggested as a new serum marker for colorectal cancer. In fact, approximately 40 percent of colorectal cancer patients have positive tumor-associated glycoprotein-72 serum levels at the time of diagnosis, while only 3 percent of patients with benign diseases are positive. A longitudinal evaluation of colorectal cancer patients suggested the utility of combining the measurement of tumor-associated glycoprotein-72 with that of carcinoembryonic antigen to monitor disease status not only at the time of diagnosis, but also at the time of recurrence. Several reports have indicated that the expression of some tumor antigens in colorectal adenomas may correlate with those parameters conventionally considered as indicative of malignant transformation. The presence of tumor-associated glycoprotein-72 in colorectal adenomas has been recently correlated with preneoplastic lesions, suggesting that tumor-associated glycoprotein-72 may be considered as an early marker of neoplastic transformation. The evaluation of tumor antigens can be considered a new tool in the management of colorectal cancer.

e21082 Background: The early diagnosis of colorectal cancers remains an area of high unmet medical need, as underscored by the U.S. estimated combined, annual death rate of >89,000. These cancers a...