Defects in MMR Genes as a Seminal Example of Personalized Medicine: From Diagnosis to Therapy

Microsatellite instability (MSI) is the landmark feature of DNA mismatch repair deficiency, which can be found in 15–20% of all colorectal cancers (CRC). This specific set of tumors has been initially perceived as a niche for geneticists or gastroenterologists focused on inherited predispositions. However, over the years, MSI has established itself as a key biomarker for the diagnosis, then extending to forecasting the disease behavior and prognostication, including the prediction of responsiveness to immunotherapy and eventually to kinase inhibitors, and possibly even to specific biological drugs. Thanks to the contribution of the characterization of MSI tumors, researchers have first acknowledged that a strong lymphocytic reaction is associated with a good prognosis. This understanding supported the prognostic implications in terms of the low metastatic potential of MSI-CRC and has led to modifications in the indications for adjuvant treatment. Furthermore, with the emergence of immunotherapy, this strong biomarker of responsiveness has exemplified the capability of re-activating an effective immune control by removing the brakes of immune evasion. Lately, a subset of MSI-CRC emerged as the ideal target for kinase inhibitors. This therapeutic scenario implies a paradox in which appropriate treatments for advanced disease are effective in a set of tumors that seldom evolve towards metastases

Prognostic and Predictive Cross-Roads of Microsatellite Instability and Immune Response to Colon Cancer

Understanding molecular features of colon cancer has shed light on its pathogenesis and progression. Over time, some of these features acquired clinical dignity and were incorporated in decision making. Namely, microsatellite instability (MSI) due to mismatch repair of defects, which primarily was adopted for the diagnosis of Lynch syndrome, became recognized as the biomarker of a different disease type, showing a less aggressive behavior. MSI tumors harbor high amounts of tumor infiltrating lymphocytes (TILs) due to their peculiar load in neoantigens. However, microsatellite stable colon cancer may also show high amounts of TILs, and this feature is as well associated with better outcomes. High TIL loads are in general associated with a favorable prognosis, especially in stage II colon cancer, and therein identifies a patient subset with the lowest probability of relapse. With respect to post-surgical adjuvant treatment, particularly in stage III, TILs predictive ability seems to weaken along with the progression of the disease, being less evident in high risk patients. Moving from cohort studies to the analysis of a series from clinical trials contributed to increase the robustness of TILs as a biomarker. The employment of high TIL densities as an indicator of good prognosis in early-stage colon cancers is strongly advisable, while in late-stage colon cancers the employment as an indicator of good responsiveness to post-surgical therapy requires refinement. It remains to be clarified whether TILs could help in identifying those patients with node-positive cancers to whom adjuvant treatment could be spared, at least in low-risk groups as defined by the TNM staging system

The impact of specialty settings on the perceived quality of medical ultrasound video

Health care professionals are increasingly viewing medical images and videos in a variety of environments. The perception of medical visual information across all specialties, career stages, and practice settings are critical to patient care and patient safety. Visual signal distortions, such as various types of noise and artifacts arising in medical imaging, affect the perceptual quality of visual content and potentially impact diagnoses. To optimize clinical practice, it is of fundamental importance to understand the way medical experts perceive visual quality. Psychophysical studies have been undertaken to evaluate the impact of visual distortions on the perceived quality of medical images and videos. However, very little research has been conducted on how speciality settings affect the perception of visual quality. In this paper, we investigate whether and how radiologists and sonographers differently perceive the quality of compressed ultrasound videos, via a dedicated subjective experiment. The findings can be used to develop useful solutions for improved visual experience and better image-based diagnoses

Quantitative evaluation of RASSF1A methylation in the non-lesional, regenerative and neoplastic liver

BACKGROUND: Epigenetic changes during ageing and their relationship with cancer are under the focus of intense research. RASSF1A and NORE1A are novel genes acting in concert in the proapoptotic pathway of the RAS signalling. While NORE1A has not been previously investigated in the human liver, recent reports have suggested that RASSF1A is frequently epigenetically methylated not only in HCC but also in the cirrhotic liver. METHODS: To address whether epigenetic changes take place in connection to age and/or to the underlying disease, we investigated RASSF1A and NORE1A gene promoter methylation by conventional methylation specific PCR and Real-Time MSP in a series of hepatitic and non-hepatitic livers harboring regenerative/hyperplastic (cirrhosis/focal nodular hyperplasia), dysplastic (large regenerative, low and high grade dysplastic nodules) and neoplastic (hepatocellular adenoma and carcinoma) growths. RESULTS: In the hepatitic liver (chronic hepatitic/cirrhosis, hepatocellular nodules and HCC) we found widespread RASSF1A gene promoter methylation with a methylation index that increased from regenerative conditions (cirrhosis) to hepatocellular nodules (p < 0.01) to HCC (p < 0.001). In the non-hepatitic liver a consistent pattern of gene methylation was also found in both lesional (focal nodular hyperplasia and hepatocellular adenoma) and non-lesional tissue. Specifically, hepatocellular adenomas (HA) showed a methylation index significantly higher than that detected in focal nodular hyperplasia (FNH) (p < 0.01) and in non-lesional tissue (p < 0.001). In non-lesional liver also the methylation index gradually increased by ageing (p = 0.002), suggesting a progressive spreading of methylated cells over time. As opposed to RASSF1A gene promoter methylation, NORE1A gene was never found epigenetically alterated in both hepatitic and non-hepatitic liver. CONCLUSION: We have shown that in non-lesional, regenerative and neoplastic liver the RASSF1A gene is increasingly methylated, that this condition takes place as an age-related phenomenon and that the early setting and spreading over time of an epigenetically methylated hepatocyte subpopulation, might be related to liver tumorigenesis

Optimization of Laboratory Diagnostics of Primary Biliary Cholangitis: When Solid-Phase Assays and Immunofluorescence Combine

The laboratory diagnostics of primary biliary cholangitis (PBC) have substantially improved, thanks to innovative analytical opportunities, such as enzyme-linked immunosorbent assays (ELISA) and multiple immunodot liver profile tests, based on recombinant or purified antigens. This study aimed to identify the best diagnostic test combination to optimize PBC diagnosis. Between January 2014 and March 2017, 164 PBC patients were recruited at the hospitals of Parma, Modena, Reggio-Emilia, and Piacenza. Antinuclear antibodies (ANA) and anti-mitochondrial antibodies (AMA) were assayed by indirect immunofluorescence (IIF), ELISA, and immunodot assays (PBC Screen, MIT3, M2, gp210, and sp100). AMA-IIF resulted in 89.6% positive cases. Using multiple immunodot liver profiles, AMA-M2 sensitivity was 94.5%, while anti-gp210 and anti-sp100 antibodies were positive in 16.5% and 17.7% of patients, respectively. PBC screening yielded positive results in 94.5% of cases; MIT3, sp100, and gp210 were detected by individual ELISA test in 89.0%, 17.1%, and 18.9% of patients, respectively. The association of PBC screening with IIF-AMA improved the diagnostic sensitivity from 89.6% to 98.2% (p &lt; 0.01). When multiple immunodot liver profile testing was integrated with AMA-IIF, the diagnostic sensitivity increased from 89.1% to 98.8% (p &lt; 0.01). The combination of IIF with solid-phase methods significantly improved diagnostic efficacy in PBC patients

Effect of extracellular matrix components on the expression of epithelial-to-mesenchymal transition markers in cultured human pancreatic ductal adenocarcinoma cells

Epithelial-to-mesenchymal transition (EMT) is a step-wise process leading to the phenotypic switch of epithelial to mesenchymal cells, providing these cells with a metastatic phenotype. During EMT epithelial cells loose adhesion by down regulation of E-cadherin and express N-cadherin, display cytoskeleton reorganization by expressing vimentin and α-smooth muscle actin (αSMA), acquire motile properties and become invasive by secretion of matrix metalloproteinases (MMPs). Cancer cell phenotype is influenced by the tumor microenvironment in relation to tumor progression, as well as to cell proliferation and invasion. The role of the extracellular matrix (ECM) in the microenvironment is particularly relevant in pancreatic ductal adenocarcinoma (PDAC) since this carcinoma is characterized by an intense desmoplastic reaction, representing the environment where the complex interplay between tumor cells, stromal fibroblasts and ECM components occurs. We aimed at analyzing in vitro the effect of the crosstalk between PDAC cells and their microenvironment by characterizing PDAC cell phenotype in cells cultured on different ECM proteins used as a substrate, in order to better understand the relationship between cancer cell behaviour and the proteins occurring in the desmoplastic tissue. We analyzed by immunofluorescence the expression of the main EMT markers such as E-cadherin, N-cadherin, β-catenin, αSMA, vimentin and collagen type I (COL-I) in PDAC cells cultured on laminin, fibronectin, COL-I and without coating (NC). Moreover, we investigated cell proliferation and MMPs activity in cell culture supernatants by SDS-zymography. Cell morphology was similar in PDAC cells cultured on laminin, fibronectin, COL-I, and in NC, as well as the E-cadherin/β-catenin complex, αSMA and COLI expression; by contrast, vimentin was undetectable in all the experimental conditions. N-cadherin was slightly detectable in cells cultured on fibronectin, COL-I, and laminin, and at lower extent in NC cells. Cell proliferation resulted similar in NC and in cells cultured on fibronectin, decreased on laminin and increased on COL-I. MMP-9 activity exhibited a similar trend, resulting similar on fibronectin, decreased on laminin and stimulated on COL-I. These preliminary results provide new insights in the characterization of the mutual effects elicited by the tumor-stroma interplay on the cancer cell, and will contribute to better understand the influence of the stroma on PDAC cancer cell phenotype, in order to develop new therapeutic strategies

Osmotic dehydration of organic kiwifruit pre-treated by pulsed electric fields and monitored by NMR

[EN] Osmotic dehydration (OD) is a widely used preservation technique that consists in the reduction in food water activity by the immersion of the biological tissue in hypertonic solutions. The aim of this work was to analyze the effect of pulsed electric fields (PEF) in mass transfer as a pre-treatment of the OD using NMR. In this sense, PEF pre-treatments were done using three different voltages (100, 250 and 400 V/cm) and 60 number of pulse. The OD of kiwifruit was carried out in 61.5% of sucrose solution at 25 °C, for a contact period from 0 to 120 min. The water distribution into the cellular tissue was studied by NMR relaxometry. In conclusion, NMR is an excellent technique for quantifying water molecules according to their interactions in the fruit tissue, obtaining the adsorbed water and opening the possibility to apply the BET model to fit the adsorbed isotherm over the whole range of water activity.The authors Urszula Tylewicz and Marco Dalla Rosa want to thank for the financial support provided by funding bodies within the FP7 ERA-Net CORE Organic Plus, and with cofounds from the European Commission. The author Maria Victoria Traffano Schiffo wants to thank the FPI Predoctoral Program of the Universidad Politecnica de Valencia for support her PhD studies, ERASMUS PRACTICAS program to finance her mobility to Italy. The authors Pedro J. Fito, Marta Castro-Giraldez and M. Victoria Traffano-Schiffo acknowledge the financial support from the Spanish Ministerio de Economia, Industria y Competitividad, Programa Estatal de I+D+i orientada a los Retos de la Sociedad AGL2016-80643-R.Traffano-Schiffo, MV.; Laghi, L.; Castro Giráldez, M.; Tylewicz, U.; Rocculi, P.; Ragni, L.; Dalla Rosa, M.... (2017). Osmotic dehydration of organic kiwifruit pre-treated by pulsed electric fields and monitored by NMR. Food Chemistry. 236:87-93. https://doi.org/10.1016/j.foodchem.2017.02.046S879323

The p50 NF-\u3baB subunit is a prognostic regulator of colorectal cancer-associated inflammation

In most tumors, tumor associated macrophages (TAMs) express an M2-skewed phenotype and are therefore associated with unfavorable prognosis. However, the impact of TAMs in colorectal cancer (CRC) development and outcome is still controversial. We first demonstrate, by parallel studies in colitis-associated cancer (CAC) and in genetically driven ApcMin mouse models, that p50 NF-\u3baB is essential for CRC development by restraining M1-dependent antitumor response. In absence of p50 mice developed fewer and smaller CRC lesions which express enhanced levels of M1/Th1 cytokines/chemokines including IL-12 and CXCL10, whose administration restrained CAC development in vivo. Moreover colons from p50-/- tumor bearers showed a reduced number of TAMs, as opposed to increased NK, NKT, CD8+ T cells and apoptotic cancer cells. Consistently, in CRC patients, high burden of p50+ TAMs was associated with decreased M1/Th1 inflammation and worse outcome indicating p50 as a new candidate for prognostic and target therapeutic intervention

Osmotic dehydration of organic kiwifruit pre-treated by pulsed electric fields: Internal transport and transformations analyzed by NMR

[EN] This work analyzes the effect of Pulsed Electric Fields (PEF) as a pre-treatment of the osmotic dehydration (OD) of kiwifruit (Actinidia deliciosa cv Hayward) in the internal structure and in the internal water transport. PEF pre-treatments were done using three PEF intensities (100, 250 and 400 V/cm) and analyzed by TD-NMR. The OD was carried out by immersing the samples in 61.5% sucrose solution at 25 °C. The application of a PEF pre-treatment before the OD produces a process of plasmolysis proportional to the electric field applied. It is because the PEF removes the mobile charges of the medium, such as electrolytes, organic acids, amino acids; Ca+2 is the major culprit of the plasmolysis because it fixes some of the junctions of the microtubules between the cell wall and the membrane. Therefore, a previous plasmolysis produces an increase in the apoplastic transport increasing the rate of dehydration.The authors Urszula Tylewicz and Marco Dalla Rosa want to thank for the financial support provided by funding bodies within the FP7 ERA-Net CORE Organic Plus, and with cofounds from the European Commission. The author Maria Victoria Traffano Schiffo wants to thank the FPI Predoctoral Program of the Universidad Politecnica de Valencia for support her PhD studies, ERASMUS PRACTICAS program to finance her mobility to Italy. The authors Pedro J. Fito, Marta Castro-Giraldez and M. Victoria Traffano-Schiffo acknowledge the financial support from the Spanish Ministerio de Economia, Industria y Competitividad, Programa Estatal de I + D + i orientada a los Retos de la Sociedad AGL2016-80643-R, Agencia Estatal de Investigacion (AEI) and Fondo Europeo de Desarrollo Regional (FEDER).Traffano-Schiffo, MV.; Laghi, L.; Castro Giráldez, M.; Tylewicz, U.; Romani, S.; Ragni, L.; Dalla Rosa, M.... (2017). Osmotic dehydration of organic kiwifruit pre-treated by pulsed electric fields: Internal transport and transformations analyzed by NMR. Innovative Food Science & Emerging Technologies. 41:259-266. https://doi.org/10.1016/j.ifset.2017.03.012S2592664