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Cáncer de mama HER2 positivo; MicroARN circulante; Tratamiento neoadyuvante Càncer de mama HER2-positiu; MicroARN circulant; Tractament neoadjuvant HER2-positive breast cancer; Circulating microRNA; Neoadjuvant treatment Background: The absence of breast cancer cells in surgical specimens, i.e., pathological complete response (pCR), is widely recognized as a favorable prognostic factor after neoadjuvant therapy. In contrast, the presence of disease at surgery characterizes a prognostically heterogeneous group of patients. Here, we challenged circulating microRNAs (miRNAs) at the end of neoadjuvant therapy as potential prognostic biomarkers in the NeoALTTO study. Methods: Patients treated within the trastuzumab arm (i.e., pre-operative weekly trastuzumab for 6 weeks followed by the addition of weekly paclitaxel for 12 weeks; post-operative FEC for 3 cycles followed by trastuzumab up to complete 1 year of treatment) were randomized into a training (n= 54) and testing (n= 72) set. RT-PCR-based high-throughput miRNA profile was performed on plasma samples collected at the end of neoadjuvant treatment of both sets. After normalization, circulating miRNAs associated with event free survival (EFS) were identified by univariate and multivariate Cox regression model. Results: Starting from 23 circulating miRNAs associated with EFS in the training set, we generated a 3-circulating miRNA prognostic signature consisting of miR-185-5p, miR-146a-5p, miR-22-3p, which was confirmed in the testing set. The 3-circulating miRNA signature showed a C-statistic of 0.62 (95% confidence interval [95%CI] 0.53-0.71) in the entire study cohort. By resorting to a multivariate Cox regression model we found a statistical significant interaction between the expression values of miR-194-5p and pCR status (p.interaction =0.005) with an estimate Hazard Ratio (HR) of 1.83 (95%CI 1.14- 2.95) in patients with pCR, and 0.87 (95%CI 0.69-1.10) in those without pCR. Notably, the model including this interaction along with the abovementioned 3-circulating miRNA signature provided the highest discriminatory capability with a C-statistic of 0.67 (95%CI 0.58-0.76). Conclusions: Circulating miRNAs are informative to identify patients with different prognosis among those with heterogeneous response after trastuzumab-based neoadjuvant treatment, and may be an exploitable tool to select candidates for salvage adjuvant therapy. The NeoALTTO study was sponsored by GlaxoSmithKline; Lapatinib is an asset of Novartis AG as of March 2, 2015. This sub-study was supported by the Italian Ministry of Health to SC. No grant number is applicable, funds were obtained through a law that allows tax-payers to allocate the 5 × 1000 share of their payments to research.

Online process control is a crucial task in modern production systems that use digital twin technology. The data acquisition from machines must provide reliable and on-the-fly data, reflecting the ...

Triple negative breast cancer (TNBC) is characterized by clinical aggressiveness, lack of recognized target therapy, and a dismal patient prognosis. Several studies addressed genomic changes occurring during neoadjuvant chemotherapy (NAC) focusing on somatic variants, but without including copy number alterations (CNAs). We analyzed CNA profiles of 31 TNBC primary tumor samples before and after NAC and of 35 single circulating tumor cells (CTCs) collected prior, during and after treatment by using next-generation sequencing targeted profile and low-pass whole genome sequencing, respectively. In pre-treatment tissue samples, the most common gains occurred on chromosomes 1, 2 and 8, and SOX11 and MYC resulted the most altered genes. Notably, amplification of MSH2 (4/4 versus 0/12, p PRDM1 and deletion of PAX3 (4/4 versus 1/12, p MYC, BCL6, SOX2, FGFR4. The phylogenetic analysis of CTCs within a single patient revealed NAC impact on tumor evolution, suggesting a selection of driver events under treatment pressure. In conclusion, our data showed how chemoresistance might arise early from treatment-induced selection of clones already present in the primary tumor, and that the characterization of CNAs on single CTCs informs on cancer evolution and potential druggable targets.

Preemptive targeted pharmacogenetic testing of candidate variations in DPYD is currently being used to limit toxicity associated with fluoropyrimidines. The use of innovative next generation sequencing (NGS) approaches could unveil additional rare (minor allele frequency 1%) genetic risk variants. However, their predictive value and management in clinical practice are still controversial, at least partly due to the challenges associated with functional analyses of rare variants. The aim of this study was to define the predictive power of rare DPYD variants burden on the risk of severe fluoropyrimidine-related toxicity. The DPYD coding sequence and untranslated regions were analyzed by NGS in 120 patients developing grade 3-5 (NCI-CTC vs3.0) fluoropyrimidine-related toxicity and 104 matched controls (no-toxicity). The functional impact of rare variants was assessed using two different in silico predictive tools (i.e., Predict2SNP and ADME Prediction Framework) and structural modeling. Plasma concentrations of uracil (U) and dihydrouracil (UH2) were quantified in carriers of the novel variants. Here, we demonstrate that the burden of rare variants was significantly higher in patients with toxicity compared to controls (p = 0.007, Mann-Whitney test). Carriers of at least one rare missense DPYD variant had a 16-fold increased risk in the first cycle and an 11-fold increased risk during the entire course of chemotherapy of developing a severe adverse event compared to controls (p = 0.013 and p = 0.0250, respectively by multinomial regression model). Quantification of plasmatic U/UH2 metabolites and in silico visualization of the encoded protein were consistent with the predicted functional effect for the novel variations. Analysis and consideration of rare variants by DPYD-sequencing could improve prevention of severe toxicity of fluoropyrimidines and improve patients' quality of life.

Background The isolation of circulating tumor cells (CTCs) requires rapid processing of the collected blood due to their inherent fragility. The ability to recover CTCs from peripheral blood mononuclear cells (PBMCs) preserved from cancer patients could allow for retrospective analyses or multicenter CTC studies. Methods We compared the efficacy of CTC recovery and characterization using cryopreserved PMBCs vs fresh whole blood from patients with non-small cell lung cancer (NSCLC; n = 8) and sarcoma (n = 6). Two epithelial cellular adhesion molecule (EpCAM)-independent strategies for CTC enrichment, based on Parsortix® technology or immunomagnetic depletion of blood cells (AutoMACS®) were tested, followed by DEPArray™ single-cell isolation. Phenotype and genotype, assessed by copy number alterations analysis, were evaluated at a single-cell level. Detection of target mutations in CTC-enriched samples from frozen NSCLC PBMCs was also evaluated by digital PCR (dPCR). Results The use of cryopreserved PBMCs from cancer patients allowed for the retrospective enumeration of CTCs and their molecular characterization, using both EpCAM-independent strategies that performed equally in capturing CTC. Cells isolated from frozen PBMCs were representative of whole blood-derived CTCs in terms of number, phenotype, and copy number aberration profile/target mutations. Long-term storage (≥3 years) did not affect the efficacy of CTC recovery. Detection of target mutations was also feasible by dPCR in CTC-enriched samples derived from stored PBMCs. Conclusions Isolating CTCs from longitudinally collected PBMCs using an unbiased selection strategy can offer a wider range of retrospective genomic/phenotypic analyses to guide patients’ personalized therapy, paving the way for sample sharing in multicenter studies.

This paper presents the experimental results obtained by applying the results of some recently concluded European projects, whose objective was the development and validation of hardware and control systems for production, based on understanding, evaluating and controlling the performance of a machine tool. It is based on a self-learning controller capable of managing a large quantity of data acquired by sensor systems, as well as on-board artifacts and finished work piece measurements that, associated to operating conditions, permit the accumulation of knowledge regarding the behaviour of machines. Relying on this experience-based approach, the controller can predict the errors that a machining process will present under different conditions and can thus adapt compensation tables. The approach set out has been implemented in a demonstrator consisting of a 5-axis high-precision boring machine, fully functional in an industrial shop floor but used under controlled environmental conditions (thermostatic chamber and special machine foundations). Its software system supports measurement procedures and is able to integrate data acquisition from different sensor systems, to calculate the volumetric error with 3D representations, to provide models for calculation of error functions and to integrate communication processes with the CNC. It can therefore operate in actual production sites, introducing relevant improvements in the machine tools manufacturing field. This paper presents the experimental results obtained during the project validation, including a comparison between on field measurements and compensation tables calculated on the basis of the predictions of the self-learning system. The analysis of the data gathered highlights the system's capability to deal with both simple linear dependencies (e.g. between error and mass variation) and complex, non-linear but repeatable trends. Results discussion, relating to two different and independent axis, demonstrates the applicability of the system under real operating conditions.

This paper is about a mechanical solution to vibration problem of a wrapper flow pack mechanism, which is used in packaging industry. The target mechanism is the welding head of the machine where the vibration is generated, due to eccentric masses rotating with a cut-on-the-fly motion law. Consequently, production rate is limited by the vibration at 200 pieces per minute. To reduce the vibration so that the production rate can exceed over 200 ppm, two ways of improvements are considered. The first one is optimizing the motion law. The other one consists in mechanical modifications which eliminates or balance out the inertia forces derived from the eccentric rotating mass. This research implements the second way of adding balance weights and discusses the properties of it. During analysis, the software ADAMS which provides kinematic simulation is used as the analytical tool to test and examine the solutions proposed.

The vibrational effects created by machines moving parts is strictly related with the characteristics of the law of motion they implement, both in case of cam-follower devices and in case of servo system electronic cam controls. One of the most effective way to prevent these harmful phenomena is to limit the harmonic content of the law of motion by designing their curve as a combination of few, low frequencies, sinusoidal curves, but this often makes difficult to satisfy other functional requirements like precision points or constant speed intervals. In this work an original method to solve this problem is illustrated. It adds to the programming language CamOMiLe new functions to assist the functional design of mechanisms. The combination of the classical theory results with the flexibility of CamOMiLe building block approach makes possible to address a large number of practical applications and two examples of them are illustrated.

Adenosine-to-Inosine (A-to-I) RNA editing is the most prevalent post-transcriptional modification of RNA molecules. Researchers have attempted to find reliable RNA editing using next generation sequencing (NGS) data. However, most of these attempts suffered from a high rate of false positives, and they did not consider the clinical relevance of the identified RNA editing, for example, in disease progression. We devised an effective RNA-editing discovery pipeline called CREDO, which includes novel statistical filtering modules based on integration of DNA- and RNA-seq data from matched tumor-normal tissues. CREDO was compared with three other RNA-editing discovery pipelines and found to give significantly fewer false positives. Application of CREDO to breast cancer data from the Cancer Genome Atlas (TCGA) project discovered highly confident RNA editing with clinical relevance to cancer progression in terms of patient survival. RNA-editing detection using DNA- and RNA-seq data from matched tumor-normal tissues should be more routinely performed as multiple omics data are becoming commonly available from each patient sample. We believe CREDO is an effective and reliable tool for this problem.

Simple Summary Distant metastases derive from the shedding and dissemination of single cancer cells (CTCs) or circulating tumor emboli (CTMs) into circulation. Previous studies on CTMs were mainly run in patients with metastatic disease; however, we observed that CTMs are more frequently detected in patients with early-stage breast cancer. Here, we collected single CTMs and their relative primary tumor tissue samples in early-stage patients. By studying genomic aberrations, present in tumors cells and absent in normal cells, we predicted the tumor fraction thanks to a statistical model developed from a calibration curve with breast cancer cell lines. The tumor fraction ranged from 8% to 48% and CTMs contained specific and shared alterations with respect to tissue. Thus, CTMs may derive from different regions of the primary tumor or from occult micrometastases. Moreover, CTM-private mutations may inform us about specific metastasis-associated functions of involved genes that should be further explored in follow-up and mechanistic studies. Abstract Circulating tumor microemboli (CTMs) are clusters of cancer cells detached from solid tumors, whose study can reveal mechanisms underlying metastatization. As they frequently comprise unknown fractions of leukocytes, the analysis of copy number alterations (CNAs) is challenging. To address this, we titrated known numbers of leukocytes into cancer cells (MDA-MB-453 and MDA-MB-36, displaying high and low DNA content, respectively) generating tumor fractions from 0–100%. After low-pass sequencing, ichorCNA was identified as the best algorithm to build a linear mixed regression model for tumor fraction (TF) prediction. We then isolated 53 CTMs from blood samples of six early-stage breast cancer patients and predicted the TF of all clusters. We found that all clusters harbor cancer cells between 8 and 48%. Furthermore, by comparing the identified CNAs of CTMs with their matched primary tumors, we noted that only 31–71% of aberrations were shared. Surprisingly, CTM-private alterations were abundant (30–63%), whereas primary tumor-private alterations were rare (4–12%). This either indicates that CTMs are disseminated from further progressed regions of the primary tumor or stem from cancer cells already colonizing distant sites. In both cases, CTM-private mutations may inform us about specific metastasis-associated functions of involved genes that should be explored in follow-up and mechanistic studies.

Simple Summary Here, we identified and characterized a claudin-low subtype of high-grade serous ovarian cancer. This rare variant of undifferentiated neoplasm shares transcriptional features with the homonym subtype of breast cancer, including low epithelial differentiation, high mesenchymal signature, and enrichment for stem cell features. Since the claudin-low transcriptional signature is associated with poor prognosis, we believe that the identification of the claudin-low molecular profile may have important clinical implications, paving the way for personalized medicine in ovarian cancer patients. Abstract Claudin-low cancer (CL) represents a rare and biologically aggressive variant of epithelial tumor. Here, we identified a claudin-low molecular profile of ovarian high-grade serous carcinoma (HGSOC), which exhibits the main characteristics of the homonym breast cancer subtype, including low epithelial differentiation and high mesenchymal signature. Hierarchical clustering and a centroid based algorithm applied to cell line collection expression dataset labeled 6 HGSOC cell lines as CL. These have a high energy metabolism and are enriched in CD44+/CD24− mesenchymal stem-like cells expressing low levels of cell-cell adhesion molecules (claudins and E-Cadherin) and high levels of epithelial-to-mesenchymal transition (EMT) induction transcription factors (Zeb1, Snai2, Twist1 and Twist2). Accordingly, the centroid base algorithm applied to large retrospective collections of primary HGSOC samples reveals a tumor subgroup with transcriptional features consistent with the CL profile, and reaffirms EMT as the dominant biological pathway functioning in CL-HGSOC. HGSOC patients carrying CL profiles have a worse overall survival when compared to others, likely to be attributed to its undifferentiated/stem component. These observations highlight the lack of a molecular diagnostic in the management of HGSOC and suggest a potential prognostic utility of this molecular subtyping.

Background As neoadjuvant chemotherapy (NAC) is increasingly used in triple-negative breast cancer (TNBC), we investigated the value of circulating tumor DNA (ctDNA) for patient monitoring prior, during, and after NAC, and circulating tumor cells (CTCs) for disease characterization at clinical progression. Materials and methods Forty-two TNBC patients undergoing NAC were prospectively enrolled. Primary tumor mutations identified by targeted-gene sequencing were validated and tracked in 168 plasma samples longitudinally collected at multiple time-points by droplet digital polymerase chain reaction. At progression, plasma DNA underwent direct targeted-gene assay, and CTCs were collected and analyzed for copy number alterations (CNAs) by low-pass whole genome sequencing. Results ctDNA detection after NAC was associated with increased risk of relapse, with 2-year event-free survival estimates being 44.4% [95% confidence interval (CI) 21.4%-92.3%] versus 77.4% (95% CI 57.8%-100%). ctDNA prognostic value remained worthy even after adjusting for age, residual disease, systemic inflammatory indices, and Ki-67 [hazard ratio (HR) 1.91; 95% CI 0.51-7.08]. During follow-up, ctDNA was undetectable in non-recurrent cases with the unique exception of one showing a temporary peak over eight samples. Conversely, ctDNA was detected in 8/11 recurrent cases, and predated the clinical diagnosis up to 13 months. Notably, recurrent cases without ctDNA developed locoregional, contralateral, and bone-only disease. At clinical progression, CTCs presented chromosome 10 and 21q CNAs whose network analysis showed connected modules including HER/PI3K/Ras/JAK signaling and immune response. Conclusion ctDNA is not only associated with but is also predictive of prognosis in TNBC patients receiving NAC, and represents an exploitable tool, either alone or with CTCs, for personalized TNBC management., Highlights • ctDNA was detected in 77% of early-stage TNBC patients undergoing neoadjuvant chemotherapy. • Patients with still detectable ctDNA after NAC were more than twice as likely to relapse as those with undetectable levels. • Detection of ctDNA during follow-up antedated clinical overt metastases up to 13 months. • ctDNA was undetectable in all but one non-recurrent patient with a temporary peak in only 1 of 8 samples tested. • CTCs of progressing cases lacked epithelial surface markers and showed therapeutically exploitable molecular features.

1093 Background: Circulating tumor cells (CTCs) expressing epithelial markers (EPCAM, cytokeratin (CK)) and lacking CD45 (a leukocyte marker) have been associated with poor outcome in many cancer types. Nonetheless, the presence of cells expressing both CK and CD45 (CK+/CD45+), circulating in the blood of cancer patients (pts) have also been reported, but not widely investigated. Early evidence indicates that circulating dual-positive cells (DPcells) are hybrids deriving from the fusion of tumor cells and macrophages. We previously reported that it is possible to detect DPcells in the blood of pts with metastatic breast cancer (BC) and that they are associated with shorter progression-free survival (PFS), in pts with pos), whereas DPcell were detected in 152 samples (44.6%, range 0-53), of which 66 (43.4%) were CTCpos and 86 (56.6%) CTCneg. Overall, DPcells were associated with a shorter OS: median OS 24.5 vs 35.0 months, p=0.046. However, when analyzing CTCpos and CTCneg separately, only the latter group showed a difference in OS according to DPcells presence. In particular, among CTCneg pts, those with ≥4 DPcells showed a 2.3-fold shorter OS (26.7 vs 60.6 months, p=0.025). Moreover, pts with ≥4 DPcells were less likely to experience a 6-months PFS clinical benefit (p=0.015). Interestingly, in the analysis by BC subtype, DPcells were confirmed to be associated with worse OS only in pts with triple negative BC (median OS 11.5 vs 16.9, p=0.048). To explore the exiology of DPcells, 2 out of 3 cells analyzed after single-cell isolation from 1 patient were confirmed to have copy number alterations (CNA) consistent with malignant cells. CNA and mutational profiling of additional single DPcells and CTCs are ongoing. Conclusions: DPcells are associated with worse OS in aBC pts, with the prognostic impact primarily in pts with

The term ‘flexural tensegrity’ applies to beam-like structures composed of segments in unilateral contact, whose integrity under flexion is provided by tendons (cables), tensioned and later anchored at the end segments. In addition to the cable tension, the constitutive response depends upon the shape of the contact surfaces between consecutive segments, identified by the corresponding pitch lines and constructed with a double couple of conjugate profiles, in order to achieve an internal constraint equivalent to a spring hinge. The response is non-local in type, because the cable elongation, and consequently the stiffness of the spring hinges, depends upon the rotations of all the segments, but this effect becomes negligible under moderate deflections. In this case, the structure can be approximated with an elastica in the continuum limit. Testing of prototypes, manufactured with a 3D printer, shows a very good agreement with the theoretical predictions for different designs of the spring hinges. The system, whose stiffness can be functionally graded and actively controlled, can be packaged when the cable is slack and deployed by pulling the cable at one extremity. It appears particularly suitable to build soft arms for robotics or deployable compliant booms for aerospace applications.

Early within-season weather conditions forecast and yield prediction can provide useful information to improve farmers' management decisions and to create a unique opportunity for implementing new solutions to specifically address key aspects of agricultural systems.Within the aims of the EU funded Horizon 2020 MED-GOLD project (https://www.med-gold.eu/), a durum wheat case study has been established to assess an innovative climate service tools for the management of climate risks and to increase yield and reduce potential risk.In this study, the added value of seasonal forecast was assessed by looking at the historical yield data and by comparing the data provided by climate service tool with traditional crop forecasting systems.For three hot spot areas (Ravenna, Ancona, and Foggia), the skills of the ECMWF-System5 seasonal time-scale forecasting provided through the Copernicus Data Store (CDS) were evaluated as a driver to the crop modeling system DELPHI, to test their added value to durum wheat yield prediction.Initially, the DELPHI model was run with observed daily weather data from sowing to harvest to obtain the reference yield. Then, yield predictions were calculated at a monthly time step, starting from February 1st and April 1st, by feeding the model with synthetic weather scenarios based on historical observations (dry, average, wet scenario - current mode) and with weather seasonal forecast (new tool) until the end of the growing season. Results for yield prediction on the basis of the current DELPHI System (historical scenarios) and on the basis of seasonal forecast (25 ensembles) were compared against reference yield.For Foggia and Ancona, in low yielding crop years and 4 months before harvest, the mean yield prediction based on the new DELPHI System tool show lower normalized root mean square error values (nRMSE) than yield predictions based on the current DELPHI system, while the latter performs better 2 months before harvest. The opposite conditions arise for the Ravenna area: lower nRMSE for the current DELPHI system 4 months before harvest and lower nRMSE for the new DELPHI system 2 months before harvest. In high yielding crop years, the new DELPHI system performs better than the current one in all the study areas both 4 and 2 months before harvest, except in Foggia where the current DELPHI system shows lower nRMSE 2 months before harvest. In general, the availability of unbiased data slightly improved the yield forecast, with the best result achieved for the high yielding crop year in Ancona, where 2 months before harvest the nRMSE dropped from 20.3% (biased) to 9.3% (unbiased). Based on these first promising results this benchmarking framework will be extended over a wider study area and for the full reanalysis temporal coverage.

Circulating tumor cells (CTCs) are promising biomarkers for prognosis, therapeutic response prediction, and treatment monitoring in cancer patients. Despite its epithelial origin, renal cell carcinoma (RCC) shows low expression of epithelial markers hindering CTC-enrichment approaches exploiting epithelial cell surface proteins. In 21 blood samples serially collected from 10 patients with metastatic RCC entering the TARIBO trial, we overcame this limitation using the marker-independent Parsortix&trade, approach for CTC-enrichment coupled with positive and negative selection with the DEPArray&trade, with single cell recovery and analysis for copy number alterations (CNA) by next generation sequencing NGS. Two CTC subpopulations were identified: epithelial CTC (eCTC) and non-conventional CTC (ncCTC) lacking epithelial and leukocyte markers. With a threshold &ge, 1CTC/10 mL of blood, the positivity rates were 28% for eCTC, 62% for ncCTCs, and 71% considering both CTC types. In two patients with detectable eCTCs at baseline, progression free survival was less than 5 months. In an index case, hierarchical structure by translational oncology (TRONCO) identified three clones among 14 CTCs collected at progression and at baseline, each containing cells with a 9p21.3loss, a well-known metastasis driving subclonal alteration. CTCs detection in RCC can be increased by marker-independent approaches, and CTC molecular characterization can allow detection of subclonal events possibly related to tumor progression.

Circulating microRNA (ct-miRNAs) are able to identify patients with differential response to HER2-targeted therapy. However, their dynamics are largely unknown. We assessed 752 miRNAs from 52 NeoALTTO patients with plasma pairs prior and two weeks after trastuzumab. Increased levels of ct-miR-148a-3p and ct-miR-374a-5p were significantly associated with pathological complete response (pCR) (p = 0.008 and 0.048, respectively). At a threshold &ge, the upper limit of the 95%CI of the mean difference, pCR resulted 45% (95%CI 24%&ndash, 68%), and 44% (95%CI 22%&ndash, 69%) for ct-miR-148a-3p and ct-miR-374a-5p, respectively. Notably, ct-miR-148a-3p retained its predictive value (OR 3.42, 95%CI 1.23&ndash, 9.46, p = 0.018) in bivariate analysis along with estrogen receptor status. Combined information from ct-miR-148a-3p and ct-miR140-5p, which we previously reported to identify trastuzumab-responsive patients, resulted in greater predictive capability over each other, with pCR of 54% (95%CI 25%&ndash, 81%) and 0% (95%CI 0%&ndash, 31%) in ct-miR-148a/ct-miR-140-5p high/present and low/absent, respectively. GO and KEGG analyses showed common enriched terms between the targets of these ct-miRNAs, including cell metabolism regulation, AMPK and MAPK signaling, and HCC progression. In conclusion, early modulated ct-miR-148-3p may inform on the functional processes underlying treatment response, integrate the information from already available predictive biomarkers, and identify patients likely to respond to single agent trastuzumab-based neoadjuvant therapy.

High-grade serous ovarian carcinoma (HGSOC) usually spreads directly into the peritoneal cavity following a transcoelomic dissemination route, although distant hematogenous metastasis exist and have been reported. However, no tumor markers can currently predict the risk of distant metastases in HGSOC. Claudins, belonging to tight-junction proteins, are dysregulated in HGSOC and functionally related to cancer progression. Here we analyzed claudin-3, -4, and -7 expression as potential markers of distant metastases. Using quantitative RT-PCR and immunohistochemistry we assessed the expression of claudins in primary HGSOC tissues, normal ovarian, and normal fallopian tube epithelia and correlated it with clinicopathological features, including the site of metastasis and the route of dissemination. Gene set enrichment analysis was performed on microarray-generated gene expression data to investigate key pathways in patients with distant metastases. We found the overall expression level of claudin-3, -4, and -7 mRNA decreased in HGSOC compared to normal tubal epithelium, currently considered the potential site of origin of many HGSOC. The reduced expression of claudin-7 is significantly associated with the development of distant metastases (p = 0.016), mainly by hematogenous route (p = 0.025). In patients with diminished expression of claudin-7, immunohistochemical staining revealed a heterogeneous pattern of membranous staining with discontinuous expression of claudin-7 along the cell border, indicative of a dischoesive architecture. The estimated reduction in the probability of distant disease is of 39% per unit increase in the level of claudin-7 (p = 0.03). Genes involved in epithelial to mesenchymal transition, hypoxia, and angiogenesis processes resulted strongly associated to hematogenous recurrence. Our data suggest a potential role of claudin-7 in discriminating distant metastatic events in HGSOC patients. The quantification of its expression levels could be a useful tool to identify patient deserving a personalized follow-up in terms of clinical and radiological assessment.

In biliary tract cancer (BTC), tissue biopsies to guide treatment are rarely feasible, thus implementing liquid biopsy approaches to improve patient management represents a priority. So far, studies on circulating tumor cells (CTCs) in BTC are insufficient to promote their use in patient clinical management and are limited to EpCAM‐enriched CTCs evaluated with the CellSearch. We applied a single‐cell protocol allowing identification not only of epithelial CTCs (eCTCs), but also of nonconventional CTCs (ncCTCs) lacking epithelial and leukocyte markers, but presenting aberrant genomes as confirmed by copy number alterations and therefore representing a distinct subpopulation of bona fide CTCs. In 41 blood samples longitudinally collected from 21 patients with advanced‐stage BTC, addition of ncCTC to classic eCTC led to a CTC‐positivity increase from 19% to 83%. Patients presenting with at least 1 eCTC/10 ml of blood at baseline prior to treatment start had a significantly shorter median disease‐specific survival (DSS) compared to those lacking eCTCs (9 months vs. 19 months, p = 0.03 by log‐rank test). No differences in DSS were observed according to ncCTC‐positivity, conversely, variations in ncCTC counts during, and at the end of treatment, were associated with the RECIST response supporting their role in treatment monitoring. Moreover, in 88 ncCTCs collected at different times during treatment, unsupervised clustering evidenced segregation of cells by patient's best response, allowing identification of genomic regions possibly involved in resistance mechanisms. The presence of ncCTCs beside eCTCs opens the way to exploiting liquid biopsy for optimizing clinical management in BTC., What's new? Late diagnosis of advanced biliary tract cancer (BTC) limits tissue biopsy for molecular analyses, resulting in missed opportunities for personalized therapy. Meanwhile, circulating tumor cells (CTCs) are promising tissue surrogates, but current CTC‐based methods detect only a fraction of BTC patients. Here, using unbiased CTC‐enrichment, coupled with identification and recovery of single cells, the authors identify a novel CTC subpopulation detectable in all BTC patient samples prior to treatment. The presence of even a single epithelial CTC was associated with reduced disease‐specific survival. This novel approach to CTC detection could be useful for treatment‐response monitoring and molecular characterization in BTC.

IoT (Internet-of-things) platforms can connect sensors and devices along supply chains of production and logistics systems, as well as end users of products, allowing for co-designed and customized solutions. This paper aims to present perspectives for the IoT-based integration of manufacturing lines. More specifically, it will address the implementation of these platforms for the cyber-physical integration of distributed and highly automated manufacturing lines of customized items. A Systematic Literature Review was conducted in order to identify the main characteristics of the research area and to cluster research and practical perspectives. As results, bibliometric analysis has evidenced the continuous growth of the research area and the most important scientific journals publishing content related to IoT-based platforms for distributed manufacturing lines. In the content analysis, the perspectives are clustered in: (i) conceptual propositions and requirements, (ii) new methods and models for supporting decision-making, (iii) development of technology-based approaches, and (iv) empirical studies. As conclusion, the paper wraps up with the description of interesting avenues from both a scientific and a praxis-oriented point-of-view.

This study presents a novel system to investigate the on-wafer level dynamic properties of GaN-based power transistors in hard-switching application conditions. The system is able to analyse devices with an on-resistance (R DSON) in the range from few ohms to hundreds of ohms, and can be effectively used to improve the development process of GaN high electron mobility transistors (HEMTs) power devices at the wafer level. Contrary to the conventional double-pulse setup, where a resistive load is usually used in combination with a very low duty cycle, the dynamic R DSON is acquired during realistic operating conditions, in a boost converter circuit. Consequently, the authors’ system is able to study not only the field-activated trapping processes, but also those induced by hard-switching conditions, i.e. promoted by hot electrons and self-heating. The maximum working voltage (600 V) and the minimum R DSON measurement time after turn-on (2 µs) allow evaluating the operation limit of the devices in a voltage/frequency range close to real switching conditions. Working on the wafer level allows a more realistic assessment of the dynamic R DSON behaviour before the packaging phase, which is very important to improve the production and development process of GaN-HEMT devices.

The capability of additive manufacturing (AM) to produce one-of-a-kind products makes it suitable to medical and pharmaceutical devices production, enabling the large-scale implementation of the personalized medicine paradigm. In this paper, to support planning, designing, and performance evaluation of an AM plant for large scale production of medical devices for healing chronic wounds, a simulation-based analysis was performed. Obtained results support planning and design decision-making, allowing for a better choice among alternative set-ups, and contribute to the evaluation of potential benefits and economic feasibility of new AM technologies.

This paper is aimed at considering supply chain and related data management within an integrated vision of the product lifecycle management (PLM) implemented through the unified approach which is proper to the Industry 4.0 initiative. In particular, with the proposed manufacturing system architecture, decision support tools can use a unified repository fed by a factory replication application, powered by data from the field, even from remote production units. Such data allow to monitor the behaviour of the digital twin of the real machine and produces a digital twin of the real product, incorporating its actual characteristics measured by means of suitable acquiring systems (in the treated example: a 3D laser scanner). Moreover, it is provided a description of the plant technological subsystems that allow to share designing and manufacturing activities across multiple similar units located in remote areas. In this context of virtual enterprise, the supply chain management results as a key factor in enabling a cooperative approach.

This paper demonstrates that—for high-electric fields and drain current levels—the electroluminescence (EL) versus VGS curves of GaN-on-Si radio frequency HEMTs significantly deviate from the well-known bell-shape, due to the turn-on of a secondary EL process that has not been described so far in the literature. Based on the combined EL measurements, electrical characterization, and thermal analysis, we demonstrate that: 1) for moderate gate and drain voltages, the EL versus VGS curve has the characteristic bell shape, and light emission originates from hot electrons injected from the source; EL signal is stronger at the edges of the gate, due to the higher electric field; 2) at high drain/gate bias and temperature, a second process induces a monotonic increase in the EL versus VGS curve; in this case, the EL signal is stronger at the center of the gate, and the EL intensity is directly correlated with the gate leakage current. Based on the experimental evidence collected within this paper, the secondary luminescence process is ascribed to: 1) the injection of carriers from the gate metal to the channel; 2) the acceleration of these electrons by the high gate–drain field; and c) the subsequent radiative emission.

Cancer treatment options are increasing. However, even among the same tumor histotype, interpatient tumor heterogeneity should be considered for best therapeutic result. Metabolomics represents the last addition to promising “omic” sciences such as genomics, transcriptomics, and proteomics. Biochemical transformation processes underlying energy production and biosynthetic processes have been recognized as a hallmark of the cancer cell and hold a promise to build a bridge between genotype and phenotype. Since breast tumors represent a collection of different diseases, understanding metabolic differences between molecular subtypes offers a way to identify new subtype-specific treatment strategies, especially if metabolite changes are evaluated in the broader context of the network of enzymatic reactions and pathways. Here, after a brief overview of the literature, original metabolomics data in a series of 92 primary breast cancer patients undergoing surgery at the Istituto Nazionale dei Tumori of Milano are reported highlighting a series of metabolic differences across various molecular subtypes. In particular, the difficult-to-treat luminal B subgroup represents a tumor type which preferentially relies on fatty acids for energy, whereas HER2 and basal-like ones show prevalently alterations in glucose/glutamine metabolism.

This paper proposes a reconfigurability index. Its development is based on five core characteristics, namely modularity, integrability, diagnosability, adaptability and customization. The index takes into consideration the interdependencies that may exist among them. The analytic network process (ANP) method is used to attribute importance weights to each core characteristic. This index can be very useful in practice since it can guide manufacturing companies to a better understanding of the various enablers of reconfigurability, as well as in the decision-making process, to decide which core characteristic requires more attention, in order to further improve the reconfigurability in existing manufacturing systems.

Triple negative breast cancer (TNBC) patients not attaining pathological Complete Response (pCR) after neo-adjuvant chemotherapy (NAC) have poor prognosis. We characterized 19 patients for somatic mutations in primary tumor biopsy and residual disease (RD) at surgery by 409 cancer-related gene sequencing (IonAmpliSeqTM Comprehensive Cancer Panel). A median of four (range 1&ndash, 66) genes was mutated in each primary tumor biopsy, and the most common mutated gene was TP53 followed by a long tail of low frequency mutations. There were no recurrent mutations significantly associated with pCR. However, half of patients with RD had primary tumor biopsy with mutations in genes related to the immune system compared with none of those achieving pCR. Overall, the number of mutations showed a downward trend in post- as compared to pre-NAC samples. PIK3CA was the most common altered gene after NAC. The mutational profile of TNBC during treatment as inferred from patterns of mutant allele frequencies in matched pre-and post-NAC samples showed that RD harbored alterations of cell cycle progression, PI3K/Akt/mTOR, and EGFR tyrosine kinase inhibitor-resistance pathways. Our findings support the use of targeted-gene sequencing for TNBC therapeutic development, as patients without pCR may present mutations of immune-related pathways in their primary tumor biopsy, or actionable targets in the RD.

3D biomaterial manufacturing strategies show an extraordinary driving force for the development of innovative therapies in the tissue engineering field. Here, the behaviour of 3D printed chitosan (CH)-based scaffolds was explored as a function of the post-printing gelation process. To this purpose, gel forming properties of different media were tested on their capability to retain 3D structure, water content, mechanical resistance and surface/internal porosity. Three different gelation media (i.e. KOH 1.5 M, Na2CO3 1.5 M, ammonia vapours) were selected and the 3D CH scaffolds were tested in terms of biocompatibility toward fibroblast as skin associated human cell line.

The present work is related to the fabrication of a scaffold with a customized shape through a variant of an additive manufacturing process often called liquid frozen deposition manufacturing, which consists in depositing layer by layer a polymer solution in a low temperature platform or chamber with an x-y-z motion platform. In this specific application, the x-y stage is actuated by high precision linear motors, located into a thermal chamber and must work at the temperature of -15° C, facing control problems related to variations of the electromagnetic interactions. Hence a multi-physics simulation has been developed in Open Modelica environment to understand the motor behavior and to allow the development of an amended control system.

The decoupling of epitaxial factors influencing on the dynamic instabilities of AlGaN/GaN metal-insulator semiconductor high electron mobility transistors is investigated. Three different sets of samples have been analyzed by means of dynamic instabilities in the threshold voltage (V$_{\mathrm{th}}$ shift). Secondary ion mass spectroscopy, steady-state photoluminescence (PL) measurements have been performed in conjunction with electrical characterization. The device dynamic performance is found to be significantly dependent on both the C concentration close to the channel as well as on the distance between the channel and the higher doped C region. Additionally, we note that experiments studying trapping should avoid large variations in the sheet carrier density (N$_{\mathrm{s}}$). This change in the N$_{\mathrm{s}}$ itself has a significant impact on the V$_{\mathrm{th}}$ shift. This experimental trends are also supported by a basic model and device simulation. Finally, the relationship between the yellow luminescence (YL) and the band edge (BE) ratio and the V$_{\mathrm{th}}$ shift is investigated. As long as the basic layer structure is not changed, the YL/BE ratio obtained from steady-state PL is demonstrated to be a valid method in predicting trap concentrations in the GaN channel layer.

The industrial world is undergoing many changes and often all these modifications are referred to as “industry 4.0”, a term introduced in Germany in 2011. There are many doubts about what is going to change within and without a company, in particular as regards the Cyber-Physical Systems (CPS) concept, and its use in this new industrial framework. Starting from the main concepts of industry 4.0 in this paper the meaning of CPS inside the framework of industry 4.0 is analyzed by defining its main features and comparing it to the business model of the fourth industrial revolution, which is based on a service oriented architecture, SOA, with the aim of a flexible, modular and customized production process.

This paper investigates the degradation of GaN-based HEMTs with p-type gate submitted to positive gate bias stress. Based on combined electrical and optical testing, we demonstrate the existence of different degradation processes, depending on the applied stress voltage ${V}_{\textsf {Gstress}}$ : 1) for ${V}_{\textsf {Gstress}} V, no significant degradation is observed, thus demonstrating a good stability of the analyzed technology; 2) for 7 V $ V, a negative shift in threshold voltage ( ${V}_{\textsf {th}}$ ) is observed, well correlated with a decrease in the gate leakage current and of the luminescence signal associated with hole injection. The negative ${V}_{\textsf {th}}$ shift is ascribed to the trapping of holes in the AlGaN and/or p-GaN/AlGaN interface; and 3) for ${V}_{\textsf {Gstress}} \ge \textsf {12}$ V, threshold voltage recovers its initial value. This is ascribed to a net-negative charge, generated either by the trapping of electrons injected from the 2-D electron gas to the AlGaN or to the de-trapping of the holes injected in 2). The results described within this paper provide relevant information for understanding the degradation dynamics of normally off GaN transistors submitted to extremely high gate voltage levels far beyond maximum use.

Through the use of experimental data and Monte Carlo simulations we investigate the shielding properties of spacecraft-shell compositions exposed to 1 GeV/nucleon 56Fe ions, representative of the worst part of the Galactic Cosmic Ray (GCR) spectrum. Through the use of the Geant4 Radiation Analysis for Space (GRAS) tool, the dose reduction and the 56Fe-fragmentation induced by those structures currently used to protect part of the International Space Station (ISS) or designed for future inflatable habitats, are analyzed. The possible effects on spacecraft electronics are discussed.

Probing electrical properties of state-of-the-art electronic devices is one of the key features of Scanning Microwave Microscopy. While providing valuable information on charge carrier properties, the combination of an atomic force microscope cantilever with a microwave signal raises the question on the actual spatial resolution of the system. On the example of the highly confined two-dimensional electron gas of an AlGaN/GaN structure, the effective tip radius is demonstrated to be in the range of the theoretical tip radius for sharp tips, while both values differ for unevenly shaped cantilever tips. The presented method demonstrates the role of the microwave excitation region for the spatial resolution of the system as well as the potential of this method to characterise the effective tip radius.

3D printers, especially in the implementation of innovative extrusion processes which do not have a long history of development, are often built by adapting mechanical designs, drives and controls previously developed for generic machine tools. This is done through a process of choice and integration which is based principally on empirical criteria and taking into account separately the different aspects and parameters. Hereafter, we present an integrated mechatronic approach which has been adopted to design from the scratch a machine to implement the innovative metal injection moulding (MIM) technology. Its extrusion rate involves the adaptation of the generated trajectories and consequently requires “ad hoc” designs, drives and numerical controls (NC) to enable non standard acceleration (and hence torque) setpoint curves. Overall, the project resulted in an acceptable workspace volume (depending on the number of degrees of freedom of the platform) and allows one to combine the extruder flow rate, the given accuracy and the required working speed (1 m/s). The system is currently used as a test bench for exploring and optimizing the parameter space of a new MIM process.

The augmented demand for medical devices devoted to tissue regeneration and possessing a controlled micro-architecture means there is a need for industrial scale-up in the production of hydrogels. A new 3D printing technique was applied to the automation of a freeze-gelation method for the preparation of chitosan scaffolds with controlled porosity. For this aim, a dedicated 3D printer was built in-house: a preliminary effort has been necessary to explore the printing parameter space to optimize the printing results in terms of geometry, tolerances and mechanical properties of the product. Analysed parameters included viscosity of the starting chitosan solution, which was measured with a Brookfield viscometer, and temperature of deposition, which was determined by filming the process with a cryocooled sensor thermal camera. Optimized parameters were applied to the production of scaffolds from solutions of chitosan alone or with the addition of raffinose as a viscosity modifier. Resulting hydrogels were characterized in terms of morphology and porosity. In vitro cell culture studies comparing 3D printed scaffolds with their homologous produced by solution casting evidenced an improvement in biocompatibility deriving from the production technique as well as from the solid state modification of chitosan stemming from the addition of the viscosity modifier.

There is nowadays a lack of knowledge and structured approaches concerning the transition of additive manufacturing systems from rapid prototyping to large scale production. The fundamental advantage of additive manufacturing to produce at virtually no additional cost complex, one-of-a-kind parts suggest addressing their integration in the direction of arranging them into the consolidated Flexible Manufacturing Systems (FMS) paradigm. In this regard, this paper presents a procedure for the simulation-based analysis of FMS hosting additive manufacturing stations, supporting their planning, design as well as performance evaluation. A discrete-event simulation model was developed and applied for the operational evaluation of an industrial case comprising 3D-printing, automated transport, and storage systems. It has been found that simulation experiments can support planning and design decision-making, allowing for a better choice among alternative set-ups. However, the present research explored an application carried out in an academic environment where the simulation model was employed for evaluating the behaviour of new additive manufacturing technologies.