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Background: Breast cancer is the most frequently diagnosed cancer in women and about 10% of breast cancer cases are hereditary. BRCA1 and BRCA2 are the genes most frequently associated with Hereditary Breast Cancer, although there are numerous other genes, such as PALB2, CHEK2 and ATM, that require to be considered as well. Germline Copy Number Variation (CNV) is one mutation type that is an important contributor to hereditary breast cancer. Nowadays, next-generation sequencing (NGS) technologies has contributed to multi-gene panel analysis used in clinical practice. Methods: In total, 1418 individuals were tested for breast cancer predisposition, using a solution-based capture approach. Targeted NGS was performed with a panel of 36 genes. The capture-based approach allowed for computational analysis of CNVs from NGS data. Results: We investigate the performance of the CNV module of the commercial software suite SeqPilot (JSI Medical Systems) and the non-commercial tool panelcn.MOPS. Both algorithms are specifically developed for CNV analysis of sequencing data reporting 99-100% sensitivity and up to 100% specificity for the prediction of CNVs up to the level of a single gene exon. All CNVs detected with these algorithms were then verified experimentally using the MLPA technique as an orthogonal assay. At least one pathogenic/likely pathogenic variant was identified in 289 samples (20.4%). CNVs accounted for 10.4% (30/289), referring to the deletion of one or more exons of a gene. Interestingly, 50% of deletions were single exon and approximately 36% of CNVs were detected in genes other than BRCA1/2. In specific, of the 30 CNVs detected, 60% occurred in BRCA1, 3.3% in BRCA2, 20% in CHEK2, 6.7% in FANCA, 6.7% in PMS2, and 3.3% in ATM. The majority of CNVs in BRCA1 were deletions of exons 19, 22, and 22-23 whereas deletions of exons 9-10 were the most common deletions in CHEK2. Detailed information of all CNVs detected is provided in Table 1. Conclusions: Our results suggest that CNV analysis should not be restricted to BRCA1/2 due to the significant proportion of CNVs (36%) in additional breast cancer predisposition genes. Furthermore, in silico CNV detection tools provide a cost-effective and feasible methodology for the identification of CNVs from NGS experiments. This outlines the clinical utility of comprehensive genetic testing that includes full sequencing and CNV analysis in hereditary breast cancer facilitating personalized management decisions for patients. Table 1.Pathogenic Copy Number Variations (CNVs) identified in this studyGeneHGVS nomenclatureOther nomenclature# detectedATMNM_000051:c.(-30+1_-29-1)_(331+1_332-1)deldeletion of exons 2-41BRCA1NM_007294:c.(5467+1_5468-1)-(*1_?)deldeletion of exon 237BRCA1NM_007294:c.(5406+1_5407-1)_(*1_?)deldeletion of exons 23-245BRCA1NM_007294:c.(5193+1_5194-1)-(5277+1_5278-1)deldeletion of exon 196BRCA2NM_000059:c.(6841+1_6842-1)_(7007+1_7008-1)deldeletion of exons 12-131CHEK2NM_007194:c.(908+1_909-1)_(1095+1_1096-1)deldeletion of exons 9-104CHEK2NM_007194:c.(792+1_793-1)_(846+1_847-1)deletion of exon 72FANCANM_000135:c.(1626+1_1627-1)_ (2852+1_2853-1)deldeletion of exons 18-291FANCANM_000135:c.(893+1_894-1)_(1359+1_1360-1)deldeletion of exons 11-141PMS2NM_000535: c.(903+1_904-1)_(988+1_989-1)deldeletion of exon 91PMS2NM_000535:c.(705+1_706-1)_(2006+1_2007-1)deldeletion of exons 7-111 Citation Format: Nikolaos Tsoulos, Konstantinos Agiannitopoulos, Georgia Pepe, Eirini Papadopoulou, Georgios N Tsaousis, Despina Apostolopoulou, Angeliki Meintani, Vassileios Venizelos, Christos Markopoulos, Rodoniki Iosifidou, Sofia Karageorgopoulou, Christos Christodoulou, Ioannis Natsiopoulos, Konstantinos Papazisis, Maria Vasilaki-Antonatou, Eleftherios Kabletsas, Amanta Psyrri, Stylianos Giassas, Dimitrios Ziogas, Efthalia Lalla, Anna Koumarianou, Christos Papadimitriou, Vahit Ozmen, Sualp Tansan, Kerim Kaban, Tahsin Ozatlı, Dan Tudor Eniu, Angelica Chiorean, Alexandru Blidaru, George Nasioulas. Different CNVs account for 10.4% of pathogenic variants in 1418 patients referred for hereditary breast cancer testing [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P2-09-10.

Background: Comprehensive tumor analysis by Next Generation Sequencing (NGS) is imperative for targeted and immunotherapy related biomarkers’ analysis in metastatic breast cancer enabling personalization of cancer treatment in these patients. In parallel with targeted therapies, immunotherapies are also evolving, revolutionizing cancer therapy. Thus, biomarkers such as. Microsatellite Instability (MSI), and Tumor Mutational Burden (TMB) are commonly employed in NGS analysis due to their correlation to Immune Checkpoint inhibitors treatment.Methods: In the present study, tumor molecular profile analysis was performed in 73 metastatic breast cancer patients using a broad NGS assay, which analyzes more than 500 unique cancer related genes and is able to detect relevant single nucleotide variations (SNVs), indels, copy number variations (CNVs) and gene fusions, in addition to TMB and MS) simultaneously. Results: An actionable alteration was detected in 79.45% of the patients. 35.62% of them had at least one alteration with an on label treatment associated and 10.96% received information related to an off-label treatment (Tiers 1A.1 and 2C.1 respectively). Additionally, 31.51% of the tumors harbored an alteration that could be used for clinical trial inclusion (Table 1). The most prevalent altered gene in these patients was the PIK3CA gene, with 27.40% mutation rate. Moreover, in 11 cases (15.07%) the mutation detected was in a gene involved in the Homologous recombination (HR) pathway, with evidence of response to PARP inhibitors. BRCA1/2 genes were as expected the most prominent HR mutated genes (5 out of 11 HR mutations), while the remaining HR mutations involved other HR genes (ATM, BLM, CHEK2, FANCM, RAD50). 24.66% of the tumors had a TMB value >10muts/MB and thus were eligible for ICI treatment. The addition of TMB to targeted biomarkers’ analysis increased the number of patients with an on-label treatment recommendation by 8.22%. Conclusions: Tumor molecular profile analysis using NGS, in real world samples, is a first-tier method for metastatic breast cancer and provides important information for decision making in the treatment of such patients. Importantly, simultaneous analysis for targeted therapy and immunotherapy biomarkers could lead to better tumor characterization and offered actionable information in 9 out of 10 patients tested. However, appropriate interpretation of results from such analysis is essential for implementation in clinical practice and accurate refinement of the treatment strategy.. Table 1.Biomarker's summary in the 73 patients included in the studyPercentage of patients with Tier1 variantsPercentage of patients with 2C.1 variantsPercentage of patients with 2C.2/2D variantsPercentage of patients with Tier 3 variantsPercentage of patients with TMB>10muts/MBMEDIAN TMBPercentage of MSI-High patients35.62% (26/73)10.96% (8/73)12.33% (9/73)0% (0/73)24.66% (18/73)5.050% (0/73) Citation Format: Eirini Papadopoulou, Nikolaos Tsoulos, Vasiliki Metaxa-Mariatou, Aikaterini Tsantikidi, Georgios Kapetsis, Chrysiida Florou-Chatzigiannidou, Sonia Maravelaki, Evgenia Bourkoula, Dimitrios Fotiou, Georgios Tsaousis, Nikolaos Touroutoglou, Dimitrios Trafalis, Ioannis Boukovinas, Ioannis Varthalitis, Zacharenia Saridaki, Charalampos Zoublios, Eleni Galani, George Papatsibas, Christos Papadimitriou, Tania Zlatintsi, Polixenia Iorga, Bülent Orhan, Sualp Tansan, Tahsin Özatlı, George Nasioulas. Comprehensive tumor analysis by NGS in metastatic breast cancer patients [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-13-01.

The recovery phase following an earthquake event is essential for urban areas with a significant number of damaged buildings. A lot of changes can take place in such a landscape within the buildings’ footprints, such as total or partial collapses, debris removal and reconstruction. Remote sensing data and methodologies can considerably contribute to site monitoring. The main objective of this paper is the change detection of the building stock in the settlement of Vrissa on Lesvos Island during the recovery phase after the catastrophic earthquake of 12 June 2017, through the analysis and processing of UAV (unmanned aerial vehicle) images and the application of Artificial Neural Networks (ANNs). More specifically, change detection of the settlement’s building stock by applying an ANN on Gray-Level Co-occurrence Matrix (GLCM) texture features of orthophotomaps acquired by UAVs was performed. For the training of the ANN, a number of GLCM texture features were defined as the independent variable, while the existence or not of structural changes in the buildings were defined as the dependent variable, assigning, respectively, the values 1 or 0 (binary classification). The ANN was trained based on the Levenberg–Marquardt algorithm, and its ability to detect changes was evaluated on the basis of the buildings’ condition, as derived from the binary classification. In conclusion, the GLCM texture feature changes in conjunction with the ANN can provide satisfactory results in predicting the structural changes of buildings with an accuracy of almost 92%.

Male breast cancer (MBC) is a very rare disorder affecting approximately 1 in 833 men. Genetic predisposition is one of the most important risk factors of MBC with BRCA2 being the most commonly mutated gene in males diagnosed with breast cancer. However, a large part of MBC heritability is still unexplained. This study sought to add to the data already available on the genetics of MBC.Our study initially involved comprehensive analysis of BRCA1 and BRCA2, followed by analysis of 43 genes implicated in cancer predisposition in a series of 100 Greek patients diagnosed with MBC between 1995-2015.Pathogenic variants were identified in 13 patients, with BRCA2 being the most commonly affected gene, followed by BRCA1, RAD50, RAD51B, and MSH3.In agreement with previous reports, BRCA2 is the most important genetic factor of MBC predisposition, while the remaining known cancer predisposition genes are each very rarely involved, rendering conclusions as to their cumulative effect difficult to draw.

A geopark can be composed of many individual geosites of various geographical scales, thus, categorization according to cartographic scale is crucial for their 3D mapping. The UNESCO Global Geopark of the island of Lesvos in the north-east Aegean, Greece, is a distinctive example of this type of geopark as it contains many unique geosites that vary in geographical scale. The geographical scale is interconnected with the cartographic scale in which the geosite is visualized. The desired cartographic scale is an essential user requirement within an unmanned aerial vehicle's (UAVs) 3D mapping project as the basis for the data acquisition strategy. This research investigates the scale issues in 3D mapping of geosites. Furthermore, it contributes to the incorporation of the geographic and cartographical scales in association with UAV flight parameters such as Ground Sample Distance (GSD), altitude, gimbal pitch, orientation, and front and side overlapping. A total of 132 geosites located in Lesvos Geopark are being studied to determine the flight parameters of three different UAVs and their camera characteristics. The methodology followed to collect very high-resolution images suitable for 3D mapping consists of five main stages: i) determining the geographical scale of each geosite, ii) defining the cartographic scale of all geosites, iii) calculation of the GSD based on cartographic scale, iv) calculation of UAV flight altitude and flight characteristics, and v) classification of geosites based on the flight characteristics for their 3D mapping. Five geographic (G1: < 0.1 ha, G2: 0.1–1 ha, G3: 1–10 ha, G4: 10–100 ha, G5: >100 ha) and five cartographic (C1: > 1:50, C2: 1:50–1:100, C3: 1:100–1:250, C4: 1:250–1:500, C5: < 1:500) categories were defined based on the geosites' size and extent. The combination of the two scales determines the most efficient flight characteristics and optimally acquires very high-resolution images required for the 3D mapping of the selected geosites. Finally, the categorization and characteristics of flights for data collection for high-resolution 3D mapping are collected and presented in a web application. The web application is addressed to the management board of Lesvos Geopark and supports the decision-making processes on mapping geosites using UAVs.

Objective Several microRNA (miRNA) polymorphisms have been associated with susceptibility to specific health disorders, including cardiovascular diseases. The aim of the present study was to investigate whether four well-studied miRNA polymorphisms in non-Caucasian populations, namely miR146a G>C (rs2910164), miR149 C>T (rs2292832), miR196a2 C>T (rs11614913) and miR499 A>G (rs3746444), contribute to the risk for the development of premature Coronary Artery Disease (CAD) in the Greek population. Methods We used a case-control study to examine these associations in 400 individuals: 200 CAD patients [including a subgroup of myocardial infraction (MI) patients] and 200 healthy controls, all of Greek origin. MiRNA polymorphisms were genotyped using three different assays: Polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP), High resolution Melting (HRM) and Sanger sequencing. Results Two of these polymorphisms, miR196a2 C>T (rs11614913) and miR499 A>G (rs3746444) were found to be strongly associated with increased risk for CAD (p=0.0388 and p=0.0013, respectively) and for MI (p=0.0281 and p=0.0273, respectively). Furthermore, miR146C-miR149C-miR196T-miR499G allele combination appeared to be significantly related to CAD (p=0.0185) and MI (p=0.0337) prevalence. Conclusions Our results suggest that at least two of the studied polymorphisms, miR196a2 C>T (rs11614913) and miR499 A>G (rs3746444), as well as the miR146C-miR149C-miR196T-miR499G allele combination could represent useful biomarkers of CAD and/or MI susceptibility in the Greek population. These special genetic characteristics, in combination with environmental factors and personal habits, might contribute to CAD and/or MI prevalence.

Background Tumor molecular profile analysis by Next Generation Sequencing technology is currently widely applied in clinical practice and has enabled the detection of predictive biomarkers of response to targeted treatment. In parallel with targeted therapies, immunotherapies are also evolving, revolutionizing cancer therapy, with Programmed Death-ligand 1 (PD-L1), Microsatellite instability (MSI), and Tumor Mutational Burden (TMB) analysis being the biomarkers employed most commonly. Methods In the present study, tumor molecular profile analysis was performed using a 161 gene NGS panel, containing the majority of clinically significant genes for cancer treatment selection. A variety of tumor types have been analyzed, including aggressive and hard to treat cancers such as pancreatic cancer. Besides, the clinical utility of immunotherapy biomarkers (TMB, MSI, PD-L1), was also studied. Results Molecular profile analysis was conducted in 610 cancer patients, while in 393 of them a at least one biomarker for immunotherapy response was requested. An actionable alteration was detected in 77.87% of the patients. 54.75% of them received information related to on-label or off-label treatment (Tiers 1A.1, 1A.2, 2B, and 2C.1) and 21.31% received a variant that could be used for clinical trial inclusion. The addition to immunotherapy biomarker to targeted biomarkers’ analysis in 191 cases increased the number of patients with an on-label treatment recommendation by 22.92%, while an option for on-label or off-label treatment was provided in 71.35% of the cases. Conclusions Tumor molecular profile analysis using NGS is a first-tier method for a variety of tumor types and provides important information for decision making in the treatment of cancer patients. Importantly, simultaneous analysis for targeted therapy and immunotherapy biomarkers could lead to better tumor characterization and offer actionable information in the majority of patients. Furthermore, our data suggest that one in two patients may be eligible for on-label ICI treatment based on biomarker analysis. However, appropriate interpretation of results from such analysis is essential for implementation in clinical practice and accurate refinement of treatment strategy.

Background Classification of splicing variants (SVs) in genes associated with hereditary cancer is often challenging. The aim of this study was to investigate the occurrence of SVs in hereditary cancer genes and the clinical utility of RNA analysis. Material and methods 1518 individuals were tested for cancer predisposition, using a Next Generation Sequencing (NGS) panel of 36 genes. Splicing variant analysis was performed using RT-PCR and Sanger Sequencing. Results In total, 34 different SVs were identified, 53% of which were classified as pathogenic or likely pathogenic. The remaining 16 variants were initially classified as Variant of Uncertain Significance (VUS). RNA analysis was performed for 3 novel variants. Conclusion The RNA analysis assisted in the reclassification of 20% of splicing variants from VUS to pathogenic. RNA analysis is essential in the case of uncharacterized splicing variants, for proper classification and personalized management of these patients.

Type 2 diabetes mellitus (DM) is a public health burden and its co-existence with hypertension is long established in the context of the metabolic syndrome. Both DM and hypertension are major risk factors, for end-stage renal disease, cardiovascular events and mortality. Strict blood pressure (BP) control in diabetics has been associated with a cardiovascular and renal risk decrease. Inhibitors of the sodium-glucose co-transporter 2 (SGLT-2) in the proximal tubule is a relatively novel class of agents for the treatment of type 2 DM. Inhibition of SGLT-2 co-transporter combines proximal tubule diuretic and osmotic diuretic action leading to glucose reabsorption reduction and mild natriuretic and diuretic effects. On this basis, several studies showed that treatment with SGLT-2 inhibitors can effectively decrease hyperglycemia but also increase BP control and reduce renal outcomes and cardiovascular mortality. Based on such evidence, the recent guidelines for the management of type 2 DM now suggest that SGLT-2 inhibitors should be preferred among oral agents in combination with metformin, in patients at increased cardiovascular risk, chronic kidney disease or heart failure. This review summarizes the existing data from studies evaluating the effect of SGLT-2 inhibitors on BP, and its potential value for cardio- and nephroprotection.

Background/Aim: The use of multi-gene panels for germline testing in breast cancer enables the estimation of cancer risk and guides risk-reducing management options. The aim of this study was to present data that demonstrate the different levels of actionability for multi-gene panels used in genetic testing of breast cancer patients and their family members. Materials and Methods: We performed an analysis in our clinical database to identify breast cancer patients undergoing genetic testing. We reviewed positive results in respect of risk estimation and management, cascade family testing, secondary findings and information for treatment decision-making. Results: A total of 415 positive test reports were identified with 57.1%, 18.1%, 10.8% and 13.5% of individuals having pathogenic/likely pathogenic variants in high, moderate, low and with insufficient evidence for breast cancer risk genes, respectively. Six point seven percent of individuals were double heterozygotes. Conclusion: Germline findings in 92% of individuals are linked to evidence-based treatment information and risk estimates for predisposition to breast and/or other cancer types. The use of germline findings for treatment decision making expands the indication of genetic testing to include individuals that could benefit from targeted treatments.

Online teaching and learning became everyday practice during the Covid-19 pandemic in education worldwide. There is a strong belief that it will remain part of common practice even in post-pandemic times. Therefore, it is important to systematically measure the user experience from the perspective of key stakeholders. In this paper, we report findings of a large scale multi-faceted usability evaluation study of tools widely used for synchronous online teaching and learning (Zoom vs competitors). The study was conducted by three independent groups of experts, who are presenting the results of the study and an analysis of the findings in this paper. The experts performed a comparative evaluation of Zoom alongside other tools used for online synchronous teaching and triangulated the findings of the study using alternative evaluation methods (personas, heuristics, interviews, etc.). The study highlights the findings of the three evaluation studies and produces a list of suggestions for improvements of the Zoom online teaching/learning platform. We also discuss the contribution of different evaluation methods in the final recommendations.

BACKGROUND NTRK gene fusions are rare in gliomas (less than 2%). The use of TRK inhibitors in NTRK-fused solid tumors has generated significant clinical interest. It is likely that the unmet need for effective therapies in glioma will lead to routine testing for NTRK fusions. METHODS We performed a retrospective review of 22 patients with malignant gliomas accrued between 2007 and 2021. We collected tumor samples from those patients and NTRK fusions were tested using either immunohistochemistry or FISH or NGS. We then evaluated their association with clinical characteristics, histology and other markers (IDH1/2 mutation, MGMT methylation, BRAF mutation, EGFR expression, ATRX expression, TERT mutation). RESULTS Median age at diagnosis was 50 years. NTRK1 translocation was detected in 3 out of 22 patient tumors, while NTRK1 duplication was present in 1 patient (probably related to a translocation, but not proven). All NTRK1 translocations were not found in glioblastomas and their median survival was 15.8 months. On the other hand, median survival of gliomas without NTRK translocations was 11 months. In one NTRK1-translocated patient a TRK inhibitor (entrectinib) was used (for a few days), but we cannot evaluate its efficacy as the patient deteriorated and died. As far as the other biomarkers are concerned, 2 out of 3 NTRK1-translocated gliomas were MGMT methylated. CONCLUSION NTRK fusions are rare in gliomas, as confirmed in our small sample analysis. Although adult NTRK-fused gliomas are considered to predominantly involve high-grade histology, in our study all patients had initially lower grade gliomas. NTRK alterations can be detected by different laboratory assays, but each of these approaches has specific advantages and limitations and more data are needed in order to identify the best method. For glioblastomas, NTRK fusions and TRK inhibitors are potentially a new targeted therapeutic strategy but more data are needed.

Ο στόχος της διατριβής είναι να μελετήσει τη σιωπή των υποκριτών σε όλες τις σωζόμενες τραγωδίες του Ευριπίδη και να καταλήξει σε συμπεράσματα σχετικά με το εύρος χρήσης της συγκεκριμένης τεχνικής και με τις διάφορες λειτουργίες που αυτή επιτελεί. Η σιωπή μελετάται από δύο σκοπιές, σκηνικά και εξωσκηνικά. Μελετώνται δηλαδή οι πραγματικές στιγμές σιωπής των υποκριτών επί σκηνής αλλά και οι αναφορές στη σιωπή. Προτείνεται μία κατηγοριοποίηση των σιωπών ανάλογα με τη λειτουργία τους και παρατίθενται όλα τα παράλληλα, τα οποία θα μπορούσαν να υποστηρίξουν τη λειτουργία αυτή. Στην κατηγορία των σκηνικών σιωπών εντάσσονται: α) η σιωπή που καθιστά τον ήρωα μη ορατό β) η σιωπή εξαιτίας ενός ισχυρού συναισθήματος γ) τα εντελώς βωβά επώνυμα και ανώνυμα πρόσωπα του δράματος του Ευριπίδη, καθώς και η ειδική κατηγορία των παιδιών δ) η κάλυψη με πέπλα. Στις εξωσκηνικές σιωπές εντάσσονται: α) η ιερή σιωπή β) το μοτίβο της παράλειψης γ) η αποσιώπηση δ) η σιωπή ως πρόληψη ατιμίας ε) η σιωπή ως ησυχία στ) ο όρκος σιωπής, είτε αυτός απευθύνεται στον χορό είτε σε επώνυμα πρόσωπα του δράματος η) η σιωπή που υποδηλώνει θάνατο και θ) η σιωπή, η οποία ανακοινώνει την είσοδο ενός ήρωα. Οι λειτουργίες της σιωπής σε όλες τις περιπτώσεις περιστρέφονται κυρίως γύρω από την επιδίωξη του τραγικού να εντείνει το δραματικό αποτέλεσμα μιας συγκεκριμένης σκηνής του δράματος ή να οργανώσει την πλοκή με έναν πιο περίτεχνο τρόπο. Παρότι πραγματική στιγμή σιωπής δεν υπάρχει, εφόσον πάντα στη σκηνή του αρχαίου δράματος κάποιος ήρωας μιλά ενώ κάποιος άλλος σιωπά, η χρήση της σιωπής δοκιμάζει τις προσδοκίες του κοινού και προκαλεί το ενδιαφέρον του. Παρατίθενται επίσης πιθανές άλλες κατηγοριοποιήσεις της σιωπής, οι οποίες πραγματοποιήθηκαν διαχρονικά από μελετητές και οι οποίες γίνονται δεκτές ή απορρίπτονται. Η έρευνα καταλήγει στο συμπέρασμα ότι ο Ευριπίδης ενέτασσε στις τραγωδίες του τη σιωπή συνειδητά και σε μεγάλη έκταση. Τελειοποίησε το δραματουργικό αυτό εργαλείο και το χρησιμοποίησε εύστοχα με στόχο να διευκολύνει την πλοκή ή να εντείνει το δραματικό αποτέλεσμα.

Σκοπός: Τα εξωκυτταρικά νουκλεϊνικά οξέα θα μπορούσαν να χρησιμοποιηθούν ως μοριακοί δείκτες στην έγκαιρη ανίχνευση του καρκίνου και στην πρόβλεψη της έκβασης της νόσου. Η παρούσα διδακτορική διατριβή επικεντρώθηκε στη μελέτη της χρησιμότητας των μοριακών δεικτών στην διάγνωση διαφόρων νεοπλασιών όπως οι καρκίνοι του προστάτη, του μαστού και του γαστρεντερικού συστήματος. Μελετήθηκαν οι εξής καρκινικοί δείκτες: 1. RNA δείκτες: a. PSMA mRNA στο αίμα ασθενών με καρκίνο του προστάτη. b. CEA mRNA στο αίμα ασθενών με νεοπλασίες του γαστρεντερικού συστήματος και του προστάτη. c. Ανίχνευση mRNA του 14-3-3σ σε ασθενείς με καρκίνους του πνεύμονα, μαστού, προστάτη και σε λευχαιμίες. 2. DNA δείκτες: a. Ποσοτικοποίηση του φορτίου DNA στο πλάσμα ασθενών με καρκίνο του προστάτη, του μαστού και του γαστρεντερικού συστήματος. b. Ανίχνευση της μεθυλίωσης στους υποκινητές των γονιδίων: I. MGMT II. 14-3-3σ III. GSTP1 IV. RASSFla V. ATM. Η μεθυλίωση στα παραπάνω γονίδια μελετήθηκε σε περιφερικό αίμα ή/και ιστούς ασθενών με καρκίνο. Μέθοδοι: To DNA πλάσμιατος ποσοτικοποιήθηκε χρησιμοποιώντας τη μέθοδο PCR πραγματικού χρόνου (Real Time PCR) καθώς και τη τυποποιημένη συσκευασία δοκιμασίας DNA DipStick™. Ελέγχθηκε επίσης η μεθυλίωση των γονιδίων MGMT, 14-3-3σ και GSTP1 χρησιμοποιώντας την ειδική για τη μεθυλίωση PCR (MéthylationSpecific PCR ή MSP), ενώ η μεθυλίωση στα γονίδια RASSF1A και του ATM στο πλάσμα ελέγχθηκε χρησιμοποιώντας τη τεχνολογία MethyLight. Η μέθοδος RT-PCR χρησιμοποιήθηκε για την ανίχνευση των PSMA και CEA mRNA. Αποτελέσματα: Οι DNA μοριακοί δείκτες ανιχνεύθηκαν σε αυξημένα ποσοστά ασθενών με καρκίνο. Σημαντικά ποσοστά ανίχνευσης παρατηρήθηκαν για τη ποσοτικοποίηση του DNA στο πλάσμα και για την μεθυλίωση στα γονίδια GSTP1, RASSF1A και ATM. Πιο συγκεκριμένα, στο 58.3% των ασθενών με πρόσφατα διαγνωσθέντα καρκίνο του προστάτη και στο 21% των ασθενών υπό θεραπεία, παρατηρήθηκε αύξηση της συγκέντρωσης του DNA στο πλάσμα. Επιπλέον, το 48.5% των ασθενών με καρκίνο του μαστού και το 62.5% των ασθενών με καρκίνο γαστρεντερικού συστήματος παρουσίασαν αυξημένες τιμές στο εξωκυτταρικό DNA.Υπερμεθυλίωση στον υποκινητή του γονιδίου GSTP1 ανιχνεύθηκε στο 75% των δειγμάτων πλάσματος που λήφθηκαν από ασθενείς με πρόσφατα διαγνωσθέντα καρκίνο του προστάτη και στο 37% των ασθενών υπό θεραπεία. Υπερμεθυλίωση στους υποκινητές των γονιδίων RASSF1A και ATM παρατηρήθηκε στο 26% και 14% αντίστοιχα των ασθενών με καρκίνο μαστού. Ανίχνευση RNA μοριακών δεικτών βρέθηκε σε μικρότερα ποσοστά στο αίμα ασθενών με καρκίνο. Συμπέρασμα: Ο συνδυασμός της μέτρησης του φορτίου DNA και της ανάλυσης της μεθυλίωσης στους υποκινητές γονιδίων θα μπορούσε να είναι μία νέα μη επεμβατική προσέγγιση για την πρώιμη ανίχνευση του καρκίνου του προστάτη και του μαστού.

Geosites are an important part of geoheritage, thus their detailed mapping is crucial for their management, protection and promotion processes. However, there is no specific approach to three-dimensional (3D) mapping of geosites and a full investigation is required, considering the current advances in the science of Geoinformatics and the need for setting up an integrated system that will suggest a suitable way of mapping areas of geological significance. The main purpose of this study is to explore new approaches to the 3D mapping of geosites, where the unmanned aerial vehicles’ (UAVs) flight planning is based on the digital elevation model (DEM). The case study that is being examined is the tectonic window of Mount Olympus, located in the southeast of Lesvos island, Greece. In this paper, a methodology has been developed to create flight plans for geosite 3D mapping. This methodology consists of three main stages: (a) flight planning based on SRTM-DEM, (b) data acquisition and image-based 3D modelling, and (c) comparison (flight plans and results). A semi-automated algorithm was developed for designing the flights, taking into account the topography of the mapped area (slope, aspect, elevation) and the final cartographic derivatives. The flight plans were compared with each other in levels of data collection, flight characteristics and their results. The results of this study are dense point clouds, DEMs and orthophotomaps. The algorithms that have been used for the comparison of point clouds were (I) surface density, (II) number of neighbours (NN), and (III) roughness and surface profile. The conclusion drawn from this study is that the DEM is a valuable source of information that can be used in designing flight plans specially shaped on the topography of each geosite.

Next-generation sequencing (NGS) technology is used to evaluate hereditary cancer risks of patients worldwide; however, information concerning the germline multigene mutational spectrum among patients with breast cancer (BC) with consanguineous marriage (CM) is limited. Therefore, this prospective study aimed to determine the molecular characteristics of patients with BC who were tested with multigene hereditary cancer predisposition NGS panel and to show the effect of CM on cancer-related genes. Patients with BC with or without CM and family history (FH) of BC treated in our breast center were selected according to The National Comprehensive Cancer Network (NCCN) criteria for hereditary BC. In these patients, the analysis of a panel of 33 genes involved in hereditary cancer predisposition was performed after genetic counseling by using NGS. The pathogenic variant (PV) and the variant of uncertain significance (VUS) were found to be 15.8 and 47.4%, respectively. PVs were identified in 10/33 genes in 34 patients; 38.2% in BRCA1/2 genes; 6, 24, and 14% in other high, moderate and low-risk genes, respectively. The CM rate was 17.7% among the 215 patients with BC. The PV rate was 13.2% in patients with CM and 16.4% in patients without CM (P=0.80). When PV and VUS were evaluated together, the PV+VUS ratio was significantly higher in patients with CM and FH of BC than patients without CM and FH of BC (88.2 vs. 63.3%, P=0.045). Analysis of multigene panel provided 9.76% additional PVs in moderate/low-risk genes. The PV rate was similar in patients with BC with or without CM. A high PV+VUS ratio in patients with CM and FH of BC suggests that genes whose importance are unknown are likely to be pathogenic genes later.

Microsatellite instability (MSI), tumor mutation burden (TMB), and programmed cell death ligand-1 (PD-L1) are particularly known as immunotherapy predictive biomarkers. MSI and TMB are closely related to DNA mismatch repair (MMR) pathway functionality, while the PD-L1 checkpoint mediates cancer cell evasion from immune surveillance via the PD-L1/PD-1 axis. Among all the novel triazolo[3,4-b]thiadiazole derivatives, the compound KA39 emerged as the most potent anticancer agent. In the present study, potential alterations in MSI, TMB, and/or PD-L1 expression upon cell treatment with KA39 are explored. We tested three MMR-deficient (DLD-1, LS174T, and DU-145) and two MMR-proficient (HT-29 and PC-3) human cancer cell lines. Our findings support KA39-induced PD-L1 overexpression in all cancer cell lines, although the most outstanding increase was observed in MMR-proficient HT-29 cells. MSI analysis showed that KA39 affects the MMR system, impairing its recognition or repair activity, particularly in MMR-deficient DLD-1 and DU-145 cells, enhancing oligonucleotide production. There were no remarkable alterations in the TMB between untreated and treated cells, indicating that KA39 does not belong to mutagenic agents. Taking together the significant in vitro anticancer activity with PD-L1 upregulation and MSI increase, KA39 should be investigated further for its implication in chemo-immunotherapy of cancer.

Background and Aims Increased blood-pressure-variability (BPV) is associated with increased cardiovascular and all-cause mortality in patients with type 2 diabetes mellitus (T2DM). Sodium-glucose co-transporter-2 (SGLT-2) inhibitors decrease the incidence of cardiovascular events, renal events, and death in this population. This study aimed to evaluate the effect of dapagliflozin on short-term BPV in patients with T2DM. Method This is a secondary analysis of a double-blind, randomized, placebo-controlled trial in 85 patients with T2DM (NCT02887677). Subjects were randomized to oral dapagliflozin 10mg once daily or placebo for 12 weeks. All participants underwent 24-h ambulatory blood pressure (BP) monitoring with the Mobil-O-Graph NG device at baseline and study-end. Standard deviation (SD), weighted SD (wSD), coefficient of variation (CV), average real variability (ARV) and variation independent of mean (VIM) were calculated with validated formulae for the 24-h and the daytime and nighttime periods. Results Dapagliflozin reduced 24-h brachial BP compared to placebo. From baseline to study-end 24-h brachial BPV indexes did not change with dapagliflozin (SBP-ARV: 11.51±3.45 vs 11.05±3.35; p=0.326, SBP-wSD: 13.59±3.60 vs 13.48±3.33; p=0.811) or placebo (SBP-ARV: 11.47±3.63 vs 11.05±3.00; p=0.388, SBP-wSD: 13.85±4.38 vs 13.97±3.87; p=0.308). Similarly, no significant changes in BPV indexes for daytime and nighttime were observed in any group. At study-end, no differences between the groups were observed for any BPV index. Deltas (Δ) of all indexes during follow-up were minimal and not different between-groups (SBP-wSD: dapagliflozin: -0.11±3.05 vs placebo: 0.12±4.20; p=0.227). Conclusion This study is the first to evaluate the effects of an SGLT-2 inhibitor on short-term BPV in patients with T2DM, showing no effect on dapagliflozin on all BPV indexes studied.

Circulating cell-free DNA (cf-DNA) is present in human biological fluids, mainly in plasma and serum, originating from cell death, a process that massively takes place during acute myocardial infarction (AMI). In the present study, cf-DNA was assessed by different quantification techniques, in order to determine its levels in patients admitted with AMI. A total of 130 subjects were included in the study: 80 ST elevation myocardial infarction (STEMI) patients and 50 healthy controls. Cf-DNA extracted from plasma was analyzed by: a) Qubit 3.0 with single (ss) and double (ds) stranded DNA assay kits, b) NanoDrop and c) quantitative PCR (qPCR). Cf-DNA levels were recorded elevated in AMI patients compared to those of healthy individuals. Specifically, Qubit 3.0 ss-DNA kit provided the highest cf-DNA concentration values for all the samples analyzed in comparison with ds-DNA assay kit and NanoDrop, approaching the values obtained by qPCR. Cf-DNA augments in massive cell death settings, including AMI, proposing that the quantification of its levels by novel methodologies could contribute to patient diagnosis and clinical management.

Fluoropyrimidine-based regimens are among the most commonly used chemotherapy combinations for the treatment of solid tumors. Several genetic polymorphisms that are implicated with fluoropyrimidine anabolism and catabolism have been associated with the development of life-threatening toxicities. Uridine triacetate is an FDA-approved antidote for 5-fluorouracil or capecitabine overdose and early-onset, life-threatening toxicity within 96 h of last chemotherapy dose. To date, it is not accessible for Greek patients as per the current summary of product characteristic's time restrictions. We report and discuss the course and outcome of capecitabine toxicity in a 66-year-old female colorectal cancer patient with heterozygous dihydropyrimidine dehydrogenase deficiency. This paper highlights the difficulty in timely access of this lifesaving medication for Greek and possibly other European patients.

Background Hereditary cancer predisposition syndromes are responsible for approximately 5–10% of all diagnosed cancer cases. In the past, single-gene analysis of specific high risk genes was used for the determination of the genetic cause of cancer heritability in certain families. The application of Next Generation Sequencing (NGS) technology has facilitated multigene panel analysis and is widely used in clinical practice, for the identification of individuals with cancer predisposing gene variants. The purpose of this study was to investigate the extent and nature of variants in genes implicated in hereditary cancer predisposition in individuals referred for testing in our laboratory. Methods In total, 1197 individuals from Greece, Romania and Turkey were referred to our laboratory for genetic testing in the past 4 years. The majority of referrals included individuals with personal of family history of breast and/or ovarian cancer. The analysis of genes involved in hereditary cancer predisposition was performed using a NGS approach. Genomic DNA was enriched for targeted regions of 36 genes and sequencing was carried out using the Illumina NGS technology. The presence of large genomic rearrangements (LGRs) was investigated by computational analysis and Multiplex Ligation-dependent Probe Amplification (MLPA). Results A pathogenic variant was identified in 264 of 1197 individuals (22.1%) analyzed while a variant of uncertain significance (VUS) was identified in 34.8% of cases. Clinically significant variants were identified in 29 of the 36 genes analyzed. Concerning the mutation distribution among individuals with positive findings, 43.6% were located in the BRCA1/2 genes whereas 21.6, 19.9, and 15.0% in other high, moderate and low risk genes respectively. Notably, 25 of the 264 positive individuals (9.5%) carried clinically significant variants in two different genes and 6.1% had a LGR. Conclusions In our cohort, analysis of all the genes in the panel allowed the identification of 4.3 and 8.1% additional pathogenic variants in other high or moderate/low risk genes, respectively, enabling personalized management decisions for these individuals and supporting the clinical significance of multigene panel analysis in hereditary cancer predisposition. Electronic supplementary material The online version of this article (10.1186/s12885-019-5756-4) contains supplementary material, which is available to authorized users.

OBJECTIVES Sodium-glucose co-transporter 2 (SGLT-2) inhibitors reduce the incidence of heart failure and death in patients with type-2 diabetes mellitus. Arterial stiffness is a prominent risk factor for heart failure and overall mortality. The aim of this study was to evaluate the effects of dapagliflozin on ambulatory brachial and central blood pressure (BP) levels and arterial stiffness parameters in patients with type-2 diabetes mellitus. METHODS This is a double-blind, randomized, placebo-controlled clinical trial including 85 adult patients with type-2 diabetes mellitus on monotherapy or combination therapy with two of: metformin, sulphonylurea, DPP-4 inhibitor, or insulin. Patients were randomized in a 1 : 1 ratio to oral dapagliflozin 10 mg per day or placebo for 12 weeks. Study participants underwent 24-h ambulatory BP monitoring with the Mobil-O-Graph NG monitor at baseline and study-end. RESULTS Baseline demographic, clinical and laboratory parameters were similar in the two groups. During follow-up, 24-h brachial SBP/DBP (129.0 ± 12.6/77.3 ± 7.3 vs. 123.2 ± 12.4/75.1 ± 6.4 mmHg; P

Background: Tumor molecular profile is of great importance for the detection of biomarkers of response to targeted treatment due to the increased availability, with concomitant reduction of cost, of Next Generation Sequencing technology (NGS). In parallel to targeted therapies’, immunotherapies are also evolving, revolutionizing cancer therapy, with Programmed Death-ligand 1 (PD-L1), Microsatellite Instability (MSI), and Tumor Mutational Burden (TMB) analysis being the biomarkers employed most commonly.Methods: In the present study, a 161 gene NGS panel, containing the majority of clinically significant genes for cancer treatment selection, was used for tumor molecular profile analysis. A variety of tumor types have been analyzed, including aggressive and hard to treat cancers such as pancreatic cancer. Besides, the clinical utility of immunotherapy biomarkers (TMB, MSI, PD-L1), was also studied.Results: Molecular profile analysis was conducted in 610 cancer patients, while in 393 of them a at least one biomarker for immunotherapy response was requested. At least one actionable alteration was detected in 77.70% of the patients. 54.59% of them received information related to on-label or off-label treatment (Tiers 1A.1, 1A.2, 2B, and 2C.1) and 21.31% received a variant that could be used for clinical trial inclusion. The addition to immunotherapy biomarker to targeted biomarkers’ analysis in 191 cases increased the number of patients with an on-label treatment recommendation by 22.40%, while an option for on-label or off-label treatment was provided in 71.35% of the cases. Conclusions: Tumor molecular profile analysis by NGS is a first-tier methodology for a variety of tumor types, which provides critical information for treatment decision making in cancer patients. Importantly, simultaneous analysis for targeted therapy and immunotherapy biomarkers could lead to a better tumor characterization and provide actionable information in the majority of patients. Moreover, our results indicate that one in two patients is eligible for ICI treatment based on the biomarkers’ analysis. However, when these analyses are performed, the challenge is their implementation in clinical practice. Multidisciplinary patient management is critical to the refinement of the strategy incorporating such information.

Background Increased blood pressure variability (BPV) is associated with increased cardiovascular and all-cause mortality in patients with type-2 diabetes mellitus (T2DM). Sodium-glucose co-transporter 2 (SGLT-2) inhibitors decrease the incidence of cardiovascular events, renal events, and death in this population. This study aimed to evaluate the effect of dapagliflozin on short-term BPV in patients with T2DM. Methods This is a secondary analysis of a double-blind, randomized, placebo-controlled trial in 85 patients with T2DM. Subjects were randomized to dapagliflozin 10 mg/day or placebo for 12 weeks. All participants underwent 24-hour ambulatory blood pressure (BP) monitoring with Mobil-O-Graph-NG device at baseline and study-end. SD, weighted SD (wSD), coefficient of variation, average real variability (ARV), and variation independent of mean were calculated for the 24-hour, daytime and nighttime periods. Results Dapagliflozin reduced 24-hour brachial BP compared with placebo. From baseline to study-end 24-hour brachial BPV indexes did not change with dapagliflozin (SBP-ARV: 11.51 ± 3.45 vs. 11.05 ± 3.35; P = 0.326, SBP-wSD: 13.59 ± 3.60 vs. 13.48 ± 3.33; P = 0.811) or placebo (SBP-ARV: 11.47 ± 3.63 vs. 11.05 ± 3.00; P = 0.388, SBP-wSD: 13.85 ± 4.38 vs. 13.97 ± 3.87; P = 0.308). Similarly, no significant changes in BPV indexes for daytime and nighttime were observed in any group. At study-end, no between-group differences were observed for any BPV index. Deltas (Δ) of all indexes during follow-up were minimal and not different between groups (SBP-wSD: dapagliflozin: −0.11 ± 3.05 vs. placebo: 0.12 ± 4.20; P = 0.227). Conclusions This study is the first to evaluate the effects of an SGLT-2 inhibitor on short-term BPV in T2DM, showing no effect of dapagliflozin on all BPV indexes studied. Clinical trials registration Trial Number NCT02887677.

Background Carriers with pathogenic variants in MSH2 have increased risk to develop colorectal, endometrium, ovarian, and other types of cancer. The PALB2 is associated with breast, ovarian, pancreatic, and prostate cancer. We describe the case of a 42‐year‐old female diagnosed with endometrial cancer at the age of 42 years with a strong family history of colorectal cancer, which was referred to our private diagnostic laboratory for genetic testing. Methods In this study, we performed next‐generation sequencing (NGS) using an amplicon based 26 genes panel. The presence of multi‐exonic copy number variations (CNVs) was investigated by computational analysis and Multiplex Ligation‐dependent Probe Amplification (MLPA). Results A gross deletion of the genomic region encompassing exons 11–16 of the MSH2 and the loss‐of‐function variant c.757_758delCT, p.(Leu253Ilefs*3) in the PALB2 were identified in the proband. Conclusions Multigene analysis using NGS technology allows the identification of pathogenic variants in genes that would normally not be tested based on the patient diagnosis. In our case these results explained not only the personal and/or family history of cancer but also allowed the surveillance for prevention of other cancer types. Moreover, the detection of large genomic rearrangements should be routinely included in hereditary cancer testing., We describe the case of a 42‐year‐old female diagnosed with endometrial cancer with a strong family history of colorectal cancer, which was referred to our private diagnostic laboratory for genetic testing. By next‐generation sequencing (NGS) and Multiplex Ligation‐dependent Probe Amplification (MLPA) analysis we identified a gross deletion of the genomic region encompassing exons 11–16 of the MSH2 gene and the loss‐of‐function variant c.757_758delCT, p.(Leu253Ilefs*3) in the PALB2 gene in our proband.