Chemical signature of colorectal cancer: case-control study for profiling the breath print.

BACKGROUND: Effective screening for colorectal cancer can reduce mortality by early detection of tumours and colonic polyps. An altered pattern of volatile organic compounds (VOCs) in exhaled breath has been proposed as a potential non-invasive diagnostic tool for detection of cancer. The aim of this study was to evaluate the reliability of breath-testing for colorectal cancer screening and early diagnosis using an advanced breath sampler. METHODS: The exhaled breath of patients with colorectal cancer and non-cancer controls with negative findings on colonoscopy was collected using the ReCIVA® Breath Sampler. This portable device is able to capture the alveolar breath fraction without environmental contamination. VOCs were desorbed thermally and analysed by gas chromatography-mass spectrometry. The discriminatory ability of VOCs in detecting colorectal cancer was evaluated by receiver operating characteristic (ROC) curve analysis for each VOC, followed by cross-validation by the leave-one-out method, and by applying stepwise logistic regression analysis. RESULTS: The study included 83 patients with colorectal cancer and 90 non-cancer controls. Fourteen VOCs were found to have significant discriminatory ability in detecting patients with colorectal cancer. The model with the diagnosis of cancer versus no cancer resulted in a statistically significant likelihood of discrimination of 173·45 (P < 0·001), with an area under the ROC curve of 0·979. Cross-validation of the model resulted in a true predictive value for colorectal cancer of 93 per cent overall. Reliability of the breath analysis was maintained irrespective of cancer stage. CONCLUSION: This study demonstrated that analysis of exhaled VOCs can discriminate patients with colorectal cancer from those without. This finding may eventually lead to the creation of a smart online sensory device, capable of providing a binary answer (cancer/no cancer) and directing to further screening.

ANTECEDENTES: Un cribaje efectivo del cáncer colorrectal (colorectal cáncer, CRC) puede reducir la mortalidad mediante la detección precoz de cáncer/pólipos del colon. La identificación de un patrón de compuestos volátiles orgánicos (volatile organic compounds, VOCs) en el aire espirado se ha propuesto como un procedimiento potencial de diagnóstico no invasivo para la detección del cáncer. El objetivo de este estudio fue evaluar la factibilidad del test de la respiración para el cribaje del CRC y diagnóstico precoz empleando un equipo avanzado de muestreo del aliento. MÉTODOS: Se recogieron muestras de aire espirado de 83 pacientes con CRC y de 90 controles sin cáncer con colonoscopia negativa empleando el ReCIVA Breath Sampler©. Este equipo portátil es capaz de capturar la fracción de aire alveolar espirada ausente de contaminación ambiental. Los VOCs fueron aislados térmicamente y analizados mediante cromatografía de gases acoplada a espectrometría de masas. La capacidad discriminatoria de los VOCs para detectar pacientes con CCR se evaluó mediante un análisis de la curva ROC para cada VOC seguida de validación cruzada mediante el método ir eliminando paso a paso cada uno de los VOCs en un modelo de regresión logística. RESULTADOS: Se observó que 14 VOCs tenían habilidad discriminatoria significativa para la detección de pacientes con CRC. El modelo con el diagnóstico de cáncer versus no cáncer mostró una probabilidad estadísticamente significativa de 151,03 (P < 0,0001) con un área bajo la curva (area under the curve, AUC) de 0,963. En la validación cruzada del modelo se obtuvo un valor global predictivo verdadero para el CRC del 92,5%. La fiabilidad del análisis del aire espirado se mantuvo con independencia del estadio del cáncer. CONCLUSIÓN: Este estudio ha demostrado que el análisis de los VOCs en el aire espirado puede discriminar pacientes con CRC de pacientes sin cáncer. Este hallazgo podría ser de ayuda para diceñar un dispositivo sensorial inteligente en línea, capaz de proporcionar una respuesta binaria (cáncer/NO cáncer) y asimismo contribuir a la indicación de una futura colonoscopia.

IN-VITRO evidence for the protective properties of the main components of the Mediterranean diet against colorectal cancer: A systematic review.

AIM: Epidemiological studies have shown that the incidence and mortality rates of colorectal cancer (CRC) vary over 10-fold worldwide where within Westernized societies lower rates are observed amongst populations living within the Mediterranean basin, suggesting a significant influence of environment and dietary style in CRC carcinogenesis. Interpretation of the data concerning the benefits of mediterranean (MD) diet is difficult in vivo because of the variability of alimentary regimens used, the differing compliance with dietary supplementation and because of the non-uniform duration of patient cohort observation. Therefore, the aim of this review is to evaluate the in-vitro effects on colorectal cancer cell lines. METHODS: the literature concerning the in-vitro effects of 4 of the principal components symbolizing the MD such as olive oil (polyphenol), red chili (capsaicin), tomato (lycopene) and red grapes (resveratrol) have been systematically reviewed. RESULTS: Several studies have demonstrated that polyphenols form olive oil, lycopene, resveratrol and capsaicin have multiple anticancer properties affecting several metabolic pathways involved in cancerogenesis, apoptosis, and metastasis in CRC cell lines. CONCLUSION: This review summarizes some of the most recent data potentially supportive of the use of MD in CRC chemoprevention, analyzing the in vitro effects of individual components of the MD on CRC cell development, progression, metastasis and apoptosis.

Fecal microRNA profile in patients with colorectal carcinoma before and after curative surgery.

PURPOSE: The purpose of this study was to explore the potential role of deranged fecal microRNA (miRNA) pattern as a reliable warning signal of colorectal cancer (CRC), a subset of fecal CRC-related miRNAs was evaluated in CRC patients, before and after surgery, and in healthy controls. METHODS: Twenty CRC patients and 20 age/sex-matched healthy volunteers with negative colonoscopy entered the study. Cancer biopsy, colonic mucosa from the resected specimens, and fecal samples from patients and controls were screened for 13 miRNAs involved in CRC onset and progressions by reverse transcription quantitative PCR (RT-qPCR). Postoperative evaluation of fecal miRNAs was carried out after a median follow-up of 18 months (range 12-30). RESULTS: Two out 13 miRNAs (RNU6B, miR-16-3p) were used as internal controls leaving 11 available for analysis. Cancer tissue contained significantly higher expression of all miRNAs, compared to normal mucosa (p < 0.05). Expression of preoperative levels of five fecal miRNAs, (miR-19-b-3p, miR-20a-5p, miR-21-3p, miR92a-3p, miR141) was significantly higher in CRC patients compared to controls and significantly decreased after curative surgery. Three out of these five miRNAs (miR20a-5p, miR21-3p, and miR141) returned to values comparable to normal controls. CONCLUSIONS: A set of three specific fecal miRNAs is overexpressed before surgery, and return within the normal range after cancer removal could be considered as an appealing opportunity for a new reliable tool for CRC secondary prevention. However, their role needs to be explored in large prospective trials and compared with the existing screening tools.

Association between follicular fluid leptin and serum insulin levels in nonoverweight women with polycystic ovary syndrome.

AIMS: We evaluated the links between leptin and visfatin levels and fertilization rates in nonoverweight (NOW) women with PCOS (NOW-PCOS) from Apulia undergoing in vitro fertilization/embryo transfer (IVF). MATERIALS AND METHODOLOGY: We recruited 16 NOW women with PCOS (NOW-PCOS) and 10 normally ovulating NOW women (control-NOW). All women underwent IVF. Androgens, 17- ß -estradiol (17 ß -E2), and insulin levels were measured in plasma and/or serum and leptin and visfatin levels were assayed in both serum and follicular fluid (FF-leptin, FF-visfatin). RESULTS: In NOW-PCOS, both serum and FF-leptin were significantly lower than in control-NOW. In NOW-PCOS, significant correlations were found between BMI and serum leptin and insulinemia and FF-leptin. By contrast, in control-NOW, FF-leptin levels were not correlated with insulinemia. Serum visfatin levels were not significantly different in NOW-PCOS and control-NOW, but FF-visfatin levels were 1.6-fold higher, although not significantly, in NOW-PCOS than in control-NOW. CONCLUSIONS: Both serum leptin levels and FF-leptin are BMI- and insulin-related in Southern Italian NOW-PCOS from Apulia. In line with other reports showing that FF-leptin levels are predictive of fertilization rates, lower than normal FF-leptin levels in NOW-PCOS may explain their lower fertilization rate and this may be related to the level of insulin and/or insulin resistance.

Disseminated tumour cells in bone marrow in experimental colon cancer: metastatic or resident?

AIM: There are conflicting data on the biological and prognostic significance of disseminated tumour cells (DTCs) in the bone marrow of colorectal cancer patients since bone metastasis is rare in this disease. The study aimed to determine the origin of bone marrow DTCs using human colorectal cancer cells in in vivo and in vitro experimental settings. METHOD: CD1 nude female mice were xenotransplanted with SW620 cells (a colorectal cancer cell line isolated from a male patient) injected in the colon wall. At autopsy, the presence of SW620 in the bone marrow (BM), colon and other organs/tissues was recognized by detection of the epithelial marker cytokeratin-19 (CK19) and Y chromosome. In addition SW620 cells or their conditioned medium were cultured with human BM cells. RESULTS: Macroscopically evident CK19+/Y-chromosome-positive tumours developed only in five mice receiving SW620 cells while putative DTCs (CK19+) were found in the bone marrow of all treated mice. Most of these CK19+ cells were Y chromosome negative, only few being Y chromosome positive. In vitro SW620 cells or their conditioned medium induced CK19 expression in cultured human bone marrow cells. CONCLUSION: Experimental colorectal cancer can induce the appearance of two distinct CK19+ cell populations in the bone marrow, one of metastatic origin and the other of murine origin. These findings suggest that bone marrow cells may undergo phenotypic modifications induced by cancer cells.

Plasma concentrations but not serum concentrations of brain-derived neurotrophic factor are related to pro-inflammatory cytokines in patients undergoing major abdominal surgery.

OBJECTIVES: To investigate peripheral brain-derived neurotrophic factor (BDNF) concentrations in the perioperative period, their relationship with transforming growth factor-ß1 (TGF-ß1 tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-6 genetics. DESIGN AND METHODS: Prospective, observational study. BDNF, TGF-ß1, IL-6 and TNF-α were analysed at baseline (T0), 5 h (T1), 24 h (T2) and 5 days (T3) after surgery, in 21 patients. The IL-6 -174 G/C polymorphism was genotyped. RESULTS: Serum BDNF concentrations decreased (P=0.048), correlated with TGF-ß1 (r=0.610 at T1, r=0.493 at T2, r=0.554 at T3). Plasma BDNF concentrations raised (P=0.049), correlated with IL-6 and TNF-α at T1 (r=0.495 and r=0.441, respectively). BDNF response was predictable from TNF-α and IL-6 concentrations and the IL-6 -174 G/C genotype. CONCLUSION: Serum and plasma BDNF concentrations could relate to platelet activation and inflammatory response, respectively. IL-6 genetics played a role in the BDNF acute response.

Anti-oestrogen therapy in the treatment of desmoid tumours: a systematic review.

AIM: The treatment of desmoid tumours (DTs) is controversial. Anti-oestrogen therapy has frequently been used, but clear information of its efficacy is lacking. In this systematic review we have undertaken a comprehensive analysis to assess the effectiveness of anti-oestrogen therapy in terms of ability to induce partial or complete regression of DTs. METHOD: A systematic review of articles published in English between January 1983 and December 2009 was carried out according to the RECIST criteria. A literature search was performed on electronic databases including: United States National Library of Medicine (MEDLINE-PubMed), Excerpta Medica (EMBASE), Cochrane Library and Google search engine. Two-hundred articles dealing with DTs were identified but only fourty-one were were selected as appropriate for the study. The chi-square test was used for statistical analysis. RESULTS: Data on 168 DTs treated with anti-oestrogen agents, alone or in combination with nonsteroidal anti-inflammatory drugs, were identified with an overall response rate of 51%. There was no difference in response according to the type of DTs or between different anti-oestrogen therapies. Combination with anti-inflammatory drugs did not improve the outcome. Toremifene was sometimes effective in cases resistant to tamoxifen. Response did not seem to be related to oestrogen receptor status. CONCLUSIONS: Despite potential inaccuracies in the methodology, the results of the review indicate that anti-oestrogen therapy produces some effect in about one half of patients with DTs. Its indication compared with other treatments is discussed.

[Stem cells and colorectal cancerogenesis: new insight]. / Cellule staminali e carcinogenesi colorettale: recenti acquisizioni.

Intestinal stem cells are monoclonal, multipotent cells residing in the niches of intestinal cripts where they regulate colonic epithelial turnover. It has been recently demonstrated that alterations in signalling transduction of the intestinal stem cells is implicated in the onset of colorectal cancer. Chronic inflammation may be one of the mechanisms involved in cancerogenesis because failure of resident stem cells in repairing the epithelial damage for the chronic insult, recruits bone marrow stem cells, which can develop genetic mutations due to the inflamed environment, leading to cancer. The main mutations associated with colorectal cancer affect the most important cellular signalling pathway, the Wnt. Mutations of adenomatous polyposis coli (APC) tumor suppressor gene and b catenin oncogene are the most common and severe alterations of this pathway. Tissue invasion and metastatization require a two-side transition of cancer stem cells, from epithelial phenotypes to mesenchimal one (epithelial transition tumor, EMT) and from the mesenchimal phenotype to the epithelial one (mesenchymal transition tumor, MET) under the regulatory effects of the environment, the intracellular b catenin distribution and P16 cell cycle inhibitor. Stem cells provide normal intestinal epithelial turnover, but may also promote intestinal cancerogenesis, and, since the cancer stem cells during the mesenchimal status are resistant to the chemotherapy (which is active only on proliferating cells), they represent the true target of future therapeutic approaches in oncology.

Tissue factor and vascular endothelial growth factor expression in colorectal cancer: relation with cancer recurrence.

OBJECTIVE: This study was undertaken to quantify tissue factor (TF) and vascular endothelial growth factor (VEGF) in colorectal cancer and to evaluate their possible relationship with recurrence. METHOD: TF and VEGF were measured by enzyme-linked immunosorbent assay in surgical tumour specimens and normal mucosa from 50 consecutive patients with colorectal cancer who were followed up for 3 years for the assessment of disease recurrence. RESULTS: TF and VEGF antigens were detected in all tumour samples. VEGF, but not TF, was much higher in tumour than in normal mucosa (P < 0.0001), as also confirmed by measurement of specific mRNAs. There was a strong correlation between TF and VEGF antigens (P < 0.0005) in tumour tissue but not in normal mucosa. Neither protein was related to tumour stage, grade or size. Local or distant recurrence was statistically related to pTNM stage. High VEGF, but not TF, levels in tumour extracts were associated with an increased risk of recurrence both by univariate (RR, 4.00, 95% CI: 1.45-11.0) and multivariate analyses (RR, 3.65, 95% CI: 1.33-10.0). CONCLUSION: These findings suggest that VEGF content in colorectal cancer is an independent risk factor for tumour recurrence and might help select patients who might benefit from adjuvant therapy.

Lipoprotein(a), lipids and proinflammatory cytokines in patients undergoing major abdominal surgery.

BACKGROUND: The aims were to investigate whether surgical stress can induce a positive or negative lipoprotein(a) acute response, to determine any association with apolipoprotein(a) phenotypes, and to establish whether any such response is dependent on changes in lipids and proinflammatory cytokines. In addition, the impact of interleukin (IL) 6 genetic variability on the cytokine response to surgery was examined. METHODS: This prospective, observational study included 41 patients with cancer referred for abdominal surgery. Preoperative (T0) plasma concentrations of lipoprotein(a), IL-6, tumour necrosis factor alpha, and serum concentrations of transforming growth factor beta1 and lipids, were compared with values obtained 5 h (T1), 24 h (T2) and 5 days (T3) after surgery. Apolipoprotein(a) Kringle IV (KIV)-VNTR (variable-number tandem repeat) and IL-6 - 174 G/C polymorphisms were analysed. RESULTS: Lipoprotein(a) was found to act as a negative acute-phase reactant (30.0 per cent reduction at T2) (P = 0.009). Surgery had a more profound impact on subjects with low KIV-VNTR. After surgery, lipoprotein(a) correlated significantly with corrected low-density lipoprotein (LDL)-cholesterol (r = 0.408 at T2). IL-6 inversely correlated with lipoprotein(a) (r = -0.321 at T1) and LDL-cholesterol (r = -0.418 at T1). The IL-6 response could be predicted from a combination of the surgical severity and -174 G/C genotype. CONCLUSION: Although temporal associations did not indicate causality, these data provide a hypothesis to explain the inverse relationship between lipoprotein(a) and IL-6.

Solutions for organ perfusion and storage: haemorheologic aspects.

BACKGROUND: The hydroxyethyl starch (HES) contained in University of Wisconsin (UW) solution causes erythrocyte aggregation. The effect of UW on red blood cell (RBC) deformability is still unclear. HES-free preservation solutions, Celsior (CS) and Custodiol (CU) are available. In this study we evaluated whether they really showed a reduced aggregating and stiffening effect on RBCs when compared with UW. We was also evaluated the effect of these solutions on cellular membranes by measuring acetylcholinesterase (AChE), which is a marker of RBC membrane integrity. METHODS: The determination of RBC aggregation and deformability was performed by a laser-assisted optical rotation cell analyzer (LORCA). AChE measurement was performed with a spectrophotometric technique. RESULTS: The mean RBC aggregation index (AI) measured in pure blood control samples was 28.00 +/- 0.73%. The AI measured samples containing UW was 38.82 +/- 1.58%. In samples with CS, it was 13.307 +/- 0.64% and in samples with CU the mean AI was 12.47 +/- 0.42%. Also the RBC aggregating time was quicker in presence of UW compared with controls. AChE concentration in blood was 3.043 +/- 0.4 nmol. CS and UW did not produce any significant change; a significant reduction was found when CU was added to blood, namely 1.975 +/- 0.1 nmol (P < .05). The use of UW or CS or CU did not result in any significant change in RBC deformability. DISCUSSION: CS and CU solutions do not aggregate erythrocytes, whereas Wisconsin does massively. CU causes an alteration of RBC cellular membrane as demonstrated by depletion of AChE.

[Expression of tissue factor and vascular endothelial growth factor in colorectal carcinoma]. / Espressione del tissue factor e del vascular endothelial growth factor nel carcinoma del colon retto.

Tissue factor (TF) and vascular endothelial growth factor (VEGF) play an important role in tumor progression and metastasis. We analyzed their expression in the carcinoma and normal mucosa of 53 colorectal cancer patients. VEGF levels were significantly higher in the tumor and correlated with TF expression. No correlation was found with tumor stage. TF may influence tumor growth and metastasis by modulating VEGF expression and neoangiogenesis.

[Gene expression of the MAGE-1, -3, and -6 antigens in hepatic carcinoma]. / Espressione genica degli antigeni MAGE-1, -3, -6 nell'epatocarcinoma.

MAGE is a family of genes specifically associated to human melanoma, but also found in hepatocellular carcinoma (HCC). In this study we evaluated the expression of such genes in 41 HCC patients, their correlation with pathological and clinical aspects of cancer, and the impact on prognosis. MAGE are expressed in most of HCC samples (28/41), no correlation was found with type and stage but they may be used as potential target for IT.