Tissue levels of oxidative stress markers and antioxidants in colorectal cancer patients

The role of reactive oxygen species in the development of cancer has become well recognized in recent years; however, evidence for a link between oxidative stress and cancer risk has not been fully explored. One of the major cancers whose number of cases has increased significantly in recent years is colon and rectal cancer, which has the second highest mortality rate in Libya. Forty subjects were divided into three groups (20 tumors from colorectal cancer patients, adjacent surrounding tumor tissues, and 20 adjacent normal tissues). Evaluation of oxidative stress indices in the samples was performed by analyzing enzymatic and non-enzymatic parameters including the activity of glutathione peroxidase and catalase as antioxidant enzymes, reduced glutathione as an antioxidant, malondialdehyde MDA levels as an oxidative damage product, nitritc oxide content NO as an inflammatory marker, and total thiols as a measure of redox status. MDA and NO levels were significantly higher in tumor tissues than in adjacent healthy tissue. Also, the surrounding tumor tissue exhibited higher MDA and NO levels compared with control tissues. The oxidant and antioxidant levels in the tumor was significantly lower than those in the surrounding tumor tissue and control healthy tissue. The results suggest that oxidant and antioxidant parameters can be used as indicators of an imbalance in humans, and as this imbalance increases, the human body may be vulnerable to developing cancer.

Overcoming Immune Depletion is the Main Goal of Developing New Therapies

In recent years, immunotherapy has revolutionized the treatment of cancer; However, inflammatory reactions in healthy tissues often have side effects that can be serious and lead to permanent discontinuation of treatment. This toxicity is not yet well understood and is a major obstacle to the use of immunotherapy. When the immune system is so severely activated, the resulting inflammatory reaction can have detrimental effects and sometimes serious damage to healthy tissue. We wanted to know if there was a difference between an optimal immune response that aims to kill cancer and an unwanted response that could affect healthy tissue. Identifying the distinctive elements between these two immune responses allows the development of new, more effective and less toxic therapeutic approaches. Keywords: Cancer; Cells; Tissues; Tumors; Prevention; Prognosis; Diagnosis; Imaging; Screening, Treatment; Management

Alpha Smooth Muscle Actin (αSMA) Immunohistochemistry Use in the Differentiation of Pancreatic Cancer from Chronic Pancreatitis

Aim: Fibrosis is observed both in pancreatic cancer (PDAC) and chronic pancreatitis (CP). The main cells involved in fibrosis are pancreatic stellate cells (PSCs), which activate alpha smooth muscle actin (αSMA), which is considered to be the best-known fibrosis marker. The aim of the study was to evaluate the expression of the αSMA in patients with PDAC and CP as the possible differentiation marker. Methods: We enrolled 114 patients undergoing pancreatic resection: 83 with PDAC and 31 with CP. Normal fragments of resected specimen from 21 patients represented the control tissue. The immunoexpressions of αSMA were detected in tissue specimens with immunohistochemistry (Abcam antibodies, GB). Results: Mean cytoplasmatic expression of αSMA protein in PDAC stromal cells was significantly higher compared to CP: 2.42 ± 0.37 vs 1.95 ± 0.45 (p < 0.01) and control group 0.61 ± 0.45 (p < 0.01). Strong immunoexpression of the αSMA protein was found in the vast majority (80.7%) of patients with PDAC, in about half (58%) of patients with CP, and not at all in healthy tissue. The expression of αSMA of different intensity was found in all patients with PDAC and CP, while in healthy tissue was minimal or absent. In PDAC patients, αSMA expression was significantly higher in tumors of diameter higher than 3 cm compared to smaller ones (p = 0.017). Conclusions: Presented findings confirm the significant role of fibrosis in both PDAC and CP; however, they do not confirm the role of αSMA as a marker of differentiation.

Investigation of Suitable Detection Angles for Carbon-Ion Radiotherapy Monitoring in Depth by Means of Secondary-Ion Tracking

The dose conformity of carbon-ion beam radiotherapy, which allows the reduction of the dose deposition in healthy tissue and the escalation of the dose to the tumor, is associated with a high sensitivity to anatomical changes during and between treatment irradiations. Thus, the monitoring of inter-fractional anatomical changes is crucial to ensure the dose conformity, to potentially reduce the size of the safety margins around the tumor and ultimately to reduce the irradiation of healthy tissue. To do so, monitoring methods of carbon-ion radiotherapy in depth using secondary-ion tracking are being investigated. In this work, the detection and localization of a small air cavity of 2 mm thickness were investigated at different detection angles of the mini-tracker relative to the beam axis. The experiments were conducted with a PMMA head phantom at the Heidelberg Ion-Beam Therapy Center (HIT) in Germany. In a clinic-like irradiation of a single field of 3 Gy (RBE), secondary-ion emission profiles were measured by a 2 cm2 mini-tracker composed of two silicon pixel detectors. Two positions of the cavity in the head phantom were studied: in front and in the middle of the tumor volume. The significance of the cavity detection was found to be increased at smaller detection angles, while the accuracy of the cavity localization was improved at larger detection angles. Detection angles of 20° – 30° were found to be a good compromise for accessing both, the detectability and the position of the air cavity along the depth in the head of a patient.

Genetic factors influence the survival rate of parotid salivary gland cancer patients

Introduction. The complex of transcriptional proteins of NF-kB (Nuclear Factor kappa-light-chain-enhancer of activated B cells) family deservedly attracts attention as a factor capable of determining the course of malignant disease. Its promising study in combination with the expression of proinflammatory gene IL6 in patients with parotid cancer (PSG) is associated with the development of modulation of malignant disease treatment and risk assessment of the disease course. Aims — to determine the effect of the expression activity of the proinflammatory interleukin-6 gene and the NFKB1 transcriptional gene on the survival rate of patients with parotid cancer. Materials and methods. A cohort retrospective study was conducted in two groups. The epidemiological group of patients included 140 people from the cancer registry of Rostov region. The clinical part of the work was carried out on 25 patients with PSG cancer of both sexes aged 50 to 80 years. Followup period of the patients after radical surgery was 18 years. Expression activity of NFKB1 and IL6 genes was estimated by real-time PCR in tumor and conditionally healthy tissue. Patient survival rate was analyzed using Kaplan-Meier method. Results. According to the results of the survival analysis in the epidemiological group, the probability that an PSG cancer patient would survive the first year after diagnosis was 95.7%, three years — 82.4%, five years — 70.9% and 10 years — 31.2%. A comparative study of gene expression levels in tumor tissue samples compared to conditionally healthy tissue revealed an increase (p<0.001) in the relative index for both the IL6 gene (5.7 times) and the NFKB1 gene (7.9 times).><0.001) in the relative index for both the IL6 gene (5.7 times) and the NFKB1 gene (7.9 times). Discussion. Analysis of our data showed the possibility of using the complex evaluation of NFKB1 and IL6 gene expression in the cells of tumor samples of PSG cancer tissue obtained during surgery to predict the long-term survival of patients after surgical treatment. Conclusions. The expression profile of NFKB1 gene in tumor tissue was a proven prognostic factor determining the course of the disease in patients with PSG cancer, which should be taken into account when forming the prognosis of the disease. The expression of IL6 gene expression in tumor cells had no independent effect on the survival rate of PSG cancer patients, but contributed to the functional activation of NFKB1 transcription gene.

Unlocking the Potential of Vaccines Built on Messenger RNA

In recent years, immunotherapy has revolutionized the treatment of cancer; however, inflammatory reactions in healthy tissues often have side effects that can be serious and lead to permanent discontinuation of treatment. This toxicity is not yet well understood and is a major obstacle to the use of immunotherapy. When the immune system is so severely activated, the resulting inflammatory reaction can have detrimental effects and sometimes serious damage to healthy tissue. We wanted to know if there was a difference between an optimal immune response that aims to kill cancer and an unwanted response that could affect healthy tissue. Identifying the distinctive elements between these two immune responses allows the development of new, more effective and less toxic therapeutic approaches. Keywords: Cancer; Cells; Tissues, Tumors; Prevention, Prognosis; Diagnosis; Imaging; Screening; Treatment; Management

95 MAGE-A1 protein expression pattern in > 5,000 tumor and healthy tissue samples: Validation of MAGE-A1 as an ideal target for TCR-based cell therapy

BackgroundCancer testis antigens (CTAs) are considered attractive targets for T cell receptor (TCR)-based cellular therapies as their expression in healthy adults is considered restricted to the immune-privileged testis. However, low-level expression of some CTAs in healthy tissue has been observed, resulting in significant on-target/off-cancer toxicity. Melanoma associated antigen 1 (MAGE-A1) is a member of the MAGE-A CTA family, whose members are known to influence cellular signaling pathways through their E3 ubiquitin ligase-binding MAGE homology domain. MAGE-A proteins are frequently expressed in different cancer types, have been linked to oncogenic activity and their expression has been associated with poor prognosis.1 Literature data suggest that in healthy tissues MAGE-A1 is detected in testis, only, with one exception suggesting MAGE-A1 RNA expression in cerebellum and cerebrum.2 Therefore, to evaluate MAGE-A1 as a potential target for cellular immunotherapies, an in-depth analysis of MAGE-A1 expression in > 70 different healthy tissue types and > 5,000 cancer biopsies was conducted, aiming to assess if MAGE-A1 represents a valid and safe target.MethodsA MAGE-A1 antibody with high specificity (TK-AbMA1P) was identified and characterized for immunohistochemistry. A large panel of > 70 different healthy tissue types and > 5,000 tumor biopsies was explored and scored for MAGE-A1 expression by tissue microarray. Identified cancer entities with relevant MAGE-A1 expression were further investigated to assess spatial intratumoral MAGE-A1 expression distribution and expression consistency between primary tumor and lymph node/distant metastases.ResultsCharacterization of TK-AbMA1P demonstrated fully paralog-selective staining for MAGE-A1. Analysis of MAGE-A1 expression in over 70 different healthy tissues confirmed strictly selective expression of MAGE-A1 in testis. An extended analysis of various CNS tissues including cerebellum and cerebrum did not reveal any expression in CNS. The analysis of > 5,000 tumor biopsies showed significant MAGE-A1 expression in distinct subgroups of multiple major tumor types with high unmet medical need. Substantial expression was detected for example in non-small-cell lung cancer, various breast cancer subtypes, gastrointestinal and urogenital cancers, among others. Extended analysis of the MAGE-A1 positive tumors demonstrated highly homogenous and consistent spatial intratumoral distribution of MAGE-A1 expression as well as between primary tumor and metastases.ConclusionsThis analysis confirms that MAGE-A1 is a highly selectively expressed CTA and demonstrates relevant expression in various indications with high unmet medical need, suggesting that MAGE-A1 is an ideal target for highly potent TCR-based adoptive cell therapy.ReferencesWeon JL, Potts PR. The MAGE protein family and cancer. Curr Opin Cell Biol 2015;37:1–8.Morgan RA, Chinnasamy N, Abate-Daga D, Gros A, Robbins PF, Zheng Z, Dudley ME, Feldman SA, Yang JC, Sherry RM, Phan GQ, Hughes MS, Kammula US, Miller AD, Hessman CJ, Stewart AA, Restifo NP, Quezado MM, Alimchandani M, Rosenberg AZ, Nath A, Wang T, Bielekova B, Wuest SC, Akula N, McMahon FJ, Wilde S, Mosetter B, Schendel DJ, Laurencot CM, Rosenberg SA. Cancer regression and neurological toxicity following anti-MAGE-A3 TCR gene therapy. J Immunother 2013;36(2):133–51.Ethics ApprovalThis study was approved by the Ethics Commission of the Ärztekammer Hamburg; approval number WF-049/09. Participants gave informed consent before taking part.

Late Gadolinium Enhancement Cardiovascular Magnetic Resonance Assessment of Substrate for Ventricular Tachycardia With Hemodynamic Compromise

Background: The majority of data regarding tissue substrate for post myocardial infarction (MI) VT has been collected during hemodynamically tolerated VT, which may be distinct from the substrate responsible for VT with hemodynamic compromise (VT-HC). This study aimed to characterize tissue at diastolic locations of VT-HC in a porcine model.Methods: Late Gadolinium Enhancement (LGE) cardiovascular magnetic resonance (CMR) imaging was performed in eight pigs with healed antero-septal infarcts. Seven pigs underwent electrophysiology study with venous arterial-extra corporeal membrane oxygenation (VA-ECMO) support. Tissue thickness, scar and heterogeneous tissue (HT) transmurality were calculated at the location of the diastolic electrograms of mapped VT-HC.Results: Diastolic locations had median scar transmurality of 33.1% and a median HT transmurality 7.6%. Diastolic activation was found within areas of non-transmural scar in 80.1% of cases. Tissue activated during the diastolic component of VT circuits was thinner than healthy tissue (median thickness: 5.5 mm vs. 8.2 mm healthy tissue, p &lt; 0.0001) and closer to HT (median distance diastolic tissue: 2.8 mm vs. 11.4 mm healthy tissue, p &lt; 0.0001). Non-scarred regions with diastolic activation were closer to steep gradients in thickness than non-scarred locations with normal EGMs (diastolic locations distance = 1.19 mm vs. 9.67 mm for non-diastolic locations, p &lt; 0.0001). Sites activated late in diastole were closest to steep gradients in tissue thickness.Conclusions: Non-transmural scar, mildly decreased tissue thickness, and steep gradients in tissue thickness represent the structural characteristics of the diastolic component of reentrant circuits in VT-HC in this porcine model and could form the basis for imaging criteria to define ablation targets in future trials.

Healthy Tissue Damage Following Cancer Ion Therapy: A Radiobiological Database Predicting Lymphocyte Chromosome Aberrations Based on the BIANCA Biophysical Model

Chromosome aberrations are widely considered among the best biomarkers of radiation health risk due to their relationship with late cancer incidence. In particular, aberrations in peripheral blood lymphocytes (PBL) can be regarded as indicators of hematologic toxicity, which is a major limiting factor of radiotherapy total dose. In this framework, a radiobiological database describing the induction of PBL dicentrics as a function of ion type and energy was developed by means of the BIANCA (BIophysical ANalysis of Cell death and chromosome Aberrations) biophysical model, which has been previously applied to predict the effectiveness of therapeutic-like ion beams at killing tumour cells. This database was then read by the FLUKA Monte Carlo transport code, thus allowing us to calculate the Relative Biological Effectiveness (RBE) for dicentric induction along therapeutic C-ion beams. A comparison with previous results showed that, while in the higher-dose regions (e.g., the Spread-Out Bragg Peak, SOBP), the RBE for dicentrics was lower than that for cell survival. In the lower-dose regions (e.g., the fragmentation tail), the opposite trend was observed. This work suggests that, at least for some irradiation scenarios, calculating the biological effectiveness of a hadrontherapy beam solely based on the RBE for cell survival may lead to an underestimation of the risk of (late) damage to healthy tissues. More generally, following this work, BIANCA has gained the capability of providing RBE predictions not only for cell killing, but also for healthy tissue damage.

Local impedance characteristics and advanced mapping capabilities to better understand pulmonary veins reconnections during repeat AF ablation procedures: insight from the CHARISMA registry

Background Detailed characterization of pulmonary veins (PV) reconnection during repeat AF ablation through high-density mapping (HDM) and local impedance (LI) algorithm is still lacking. Purpose We aimed to characterize PV gaps and underlying electrical activity during and after ablation of PVs in AF patients (pts). Methods Consecutive patients (pts) undergoing redo AF ablation from the CHARISMA registry with complete characterization of PV gaps (PVG) at 8 Italian centers were included. Rhythmia mapping system was used to map the left atrium and PVs before and after ablation. LI characteristics were collected through a RF ablation catheter equipped with a dedicated LI algorithm (DirectSense). A novel map analysis tool (Lumipoint) that automatically identifies split potentials and continuous activation was used sequentially on each PV component, in order to better assess PVG. Each PVG was characterized in terms of LI and its variations during the procedure. Ablation endpoint was PVI as assessed by entrance and exit block. Results Fifty PVGs were automatically identified through the Lumipoint tool in 23 cases, mostly at anterior sites (21, 42%), followed by posterior (15, 30%) and carina (10, 20%) sites. One PVG was identified in 7 (28%) pts, 2 gaps in 10 (43.5%) pts and &gt;2 gaps in 6 (26.1%) pts. The mean LI at PVG sites was 111.3±12Ω prior to ablation: it was significantly higher than LI at scar tissue closer to PVG (99.3±8Ω, p&lt;0.0001) but was significantly lower than LI at healthy tissue (120.8±11Ω, p=0.0015). The mean linear extension of PVGs detected through Lumipoint was significantly lower than the one recognized through voltage map (11.5±8 mm vs 13.3±9 mm, p=0.01) whereas was comparable to the one identified through conventional activation map (11.8±7 mm, p=0.1161 vs Lumipoint). Complete identification of the whole area of PVG was achieved in 31 (62%) and 42 (84%) cases through voltage and activation map, respectively whereas the identification was only partial in 18 (36%) and 7 (14%) cases, respectively. In 1 case both voltage and activation map failed to identify a PVG. No complications during the procedures were reported. All PVs were successfully isolated in all study pts. Conclusion Advanced mapping capabilities were useful to pinpoint the search for PVGs, enabling a more targeted ablation approach vs relying on voltage mapping. LI values correlated well with PVGs characteristics and they significantly differ from both scar and healthy tissue. FUNDunding Acknowledgement Type of funding sources: None.