Search

Pancreatic ductal adenocarcinoma (PDA) has been predicted to soon becomethe second leading cause of cancer related deaths. Although poly (ADPribose)polymerase (PARP) inhibitors (PARPi) have emerged as promising anticancerdrugs for BRCA mutated tumors, in pancreatic cancer, phase III clinical resultshave failed. Major limitations of PARPi are due to their lack of selectivity. In thisregard, we have recently demonstrated that Parp-2, but not Parp-1, plays aspecific role in replicative stress, which is a known PDA hallmark. Therefore, wehypothesized that Parp-2 may represent a new potential target to fight againstPDA. Importantly, Parp-2 genetic deletion in Ela-myc transgenic mice resulted in a43% increase in animal survival. Tumor histopathological characterizationshowed that Parp-2 inhibition hampers acinarto-ductal metaplasia, a key eventin pancreatic cancer initiation. Parp-2 loss in vitro significantly decreasedtumoroid generation capacity and induced DNA damage accumulation,increasing replicative stress and apoptosis. Molecular analysis of Ela-myc:Parp-2-/- vs Ela-myc Parp-2+/+ pancreatic tumors in preneoplastic lesions, indicatedthat Parp-2 inhibition resulted in a less immune-evasive environment. Moreover,GSEA at late tumor stages shows p53 pathway enrichment in Ela-myc Parp-2+/+vs Ela-myc:Parp-2-/-. Indeed, p53 inactivation was found in primary cell linesestablished from late stage Ela-myc:Parp-2-/- tumors, suggesting that loss offunction of this pathway is required for tumor progression in the absence ofParp-2. Altogether our data highlight that Parp-2 is a novel target in pancreaticcancer and open new avenues for therapeutic intervention against thisaggressive tumor by using specific Parp-2 pharmacological inhibitors.

Body thermoregulation during exercise induces sweating with the consequent loss of water, electrolytes and other compounds. The use of sweat for bioanalytical purposes has recently widespread because it is an easily accessible biofluid that can be noninvasively collected and/or directly monitored using wearable devices. Different sweat biomarkers can be monitored as indicators of the physiological status of an individual. The concentration of sweat Na+ electrolyte provides information about dehydration or hyponatremia events. Among the different sensor technologies applied for Na+ ion detection in sweat, ion-sensitive field-effect transistors (ISFETs) show superior features in terms miniaturization, key for working with the low volumes of sweat available, robustness, scalability and reproducibility. They also offer fast response and low impedance output signal. This work reports an in-depth study that thoroughly assesses the potential of ISFET sensors for sweat Na+ analysis. Results show a reproducible sensitivity of 60.7 ± 0.5 mV (-Log aNa)− 1, high repeatability, and lifetime up to one month. Sensor reliability is demonstrated by analysing 20 sweat samples and results are compared with the ones provided with the standard ion chromatography technique (IC). The statistical analysis demonstrates that Na+ concentrations estimated with the ISFET sensor and IC are in good agreement showing a relative error of up to 20%. Results demonstrate that the sensor presented in this work can potentially be used for the continuous monitoring of Na+ changes in sweat during exercise.

Sweat secreted by the human eccrine sweat glands can provide valuable biomarker information during exercise. Real-time non-invasive biomarker recordings are therefore useful for evaluating the physiological conditions of an athlete such as their hydration status during endurance exercise. This work describes a wearable sweat biomonitoring patch incorporating printed electrochemical sensors into a plastic microfluidic sweat collector and data analysis that shows the real-time recorded sweat biomarkers can be used to predict a physiological biomarker. The system was placed on subjects carrying out an hour-long exercise session and results were compared to a wearable system using potentiometric robust silicon-based sensors and to commercially available HORIBA-LAQUAtwin devices. Both prototypes were applied to the real-time monitoring of sweat during cycling sessions and showed stable readings for around an hour. Analysis of the sweat biomarkers collected from the printed patch prototype shows that their real-time measurements correlate well (correlation coefficient ≥ 0.65) with other physiological biomarkers such as heart rate and regional sweat rate collected in the same session. We show for the first time, that the real-time sweat sodium and potassium concentration biomarker measurements from the printed sensors can be used to predict the core body temperature with root mean square error (RMSE) of 0.02 °C which is 71% lower compared to the use of only the physiological biomarkers. These results show that these wearable patch technologies are promising for real-time portable sweat monitoring analytical platforms, especially for athletes performing endurance exercise.

Pancreatic ductal adenocarcinoma (PDA) has been predicted to soon become the second leading cause of cancer related deaths. Although poly (ADP-ribose) polymerase (PARP) inhibitors (PARPi) have emerged as promising anti-cancer drugs for BRCA mutated tumors, in pancreatic cancer, phase III clinical results have failed. Major limitations of PARPi are due to their lack of selectivity. In this regard, we have recently demonstrated that Parp-2, but not Parp-1, plays a specific role in replicative stress, which is a known PDA hallmark. Therefore, we hypothesized that Parp-2 may represent a new potential target to fight against PDA. Importantly, Parp-2 genetic deletion in Ela-myc transgenic mice resulted in a 43% increase in animal survival. Tumor histopathological characterization showed that Parp-2 inhibition hampers acinar-to- ductal metaplasia, a key event in pancreatic cancer initiation. Parp-2 loss in vitro significantly decreased tumoroid generation capacity and induced DNA damage accumulation, increasing replicative stress and apoptosis. Molecular analysis of Ela-myc:Parp-2-/- vs Ela-myc Parp- 2+/+ pancreatic tumors in preneoplastic lesions, indicated that Parp-2 inhibition resulted in a less immune-evasive environment. Moreover, GSEA at late tumor stages shows p53 pathway enrichment in Ela-myc Parp-2+/+ vs Ela-myc:Parp-2-/-. Indeed, p53 inactivation was found in primary cell lines established from late stage Ela-myc:Parp-2-/- tumors, suggesting that loss of function of this pathway is required for tumor progression in the absence of Parp-2. Altogether our data highlight that Parp-2 is a novel target in pancreatic cancer and open new avenues for therapeutic intervention against this aggressive tumor by using specific Parp-2 pharmacological inhibitors.

Body thermoregulation during exercise induces sweating with the consequent loss of water, electrolytes and other compounds. The use of sweat for bioanalytical purposes has recently widespread because it is an easily accessible biofluid that can be noninvasively collected and/or directly monitored using wearable devices. Different sweat biomarkers can be monitored as indicators of the physiological status of an individual. The concentration of sweat Na+ electrolyte provides information about dehydration or hyponatremia events. Among the different sensor technologies applied for Na+ ion detection in sweat, ion-sensitive field-effect transistors (ISFETs) show superior features in terms miniaturization, key for working with the low volumes of sweat available, robustness, scalability and reproducibility. They also offer fast response and low impedance output signal. This work reports an in-depth study that thoroughly assesses the potential of ISFET sensors for sweat Na+ analysis. Results show a reproducible sensitivity of 60.7 ± 0.5 mV (-Log aNa)− 1, high repeatability, and lifetime up to one month. Sensor reliability is demonstrated by analysing 20 sweat samples and results are compared with the ones provided with the standard ion chromatography technique (IC). The statistical analysis demonstrates that Na+ concentrations estimated with the ISFET sensor and IC are in good agreement showing a relative error of up to 20%. Results demonstrate that the sensor presented in this work can potentially be used for the continuous monitoring of Na+ changes in sweat during exercise.

Sweat secreted by the human eccrine sweat glands can provide valuable biomarker information during exercise. Real-time non-invasive biomarker recordings are therefore useful for evaluating the physiological conditions of an athlete such as their hydration status during endurance exercise. This work describes a wearable sweat biomonitoring patch incorporating printed electrochemical sensors into a plastic microfluidic sweat collector and data analysis that shows the real-time recorded sweat biomarkers can be used to predict a physiological biomarker. The system was placed on subjects carrying out an hour-long exercise session and results were compared to a wearable system using potentiometric robust silicon-based sensors and to commercially available HORIBA-LAQUAtwin devices. Both prototypes were applied to the real-time monitoring of sweat during cycling sessions and showed stable readings for around an hour. Analysis of the sweat biomarkers collected from the printed patch prototype shows that their real-time measurements correlate well (correlation coefficient ≥0.65 ) with other physiological biomarkers such as heart rate and regional sweat rate collected in the same session. We show for the first time, that the real-time sweat sodium and potassium concentration biomarker measurements from the printed sensors can be used to predict the core body temperature with root mean square error (RMSE) of 0.02 °C which is 71% lower compared to the use of only the physiological biomarkers. These results show that these wearable patch technologies are promising for real-time portable sweat monitoring analytical platforms, especially for athletes performing endurance exercise.

Acknowledgments We thank the Microscopy, Cytometry, Comparative Pathology, Mouse Facility, and Mouse Genome editing core services of the CNIO for their support. We also thank Eva Porlan (UAM, Madrid) for help with neural cultures, and Dr. Mario García‐Domínguez (CABIMER, Seville) for the UTF1 3KR and 5KR mutants. CV received support from the Juan de la Cierva programme, Ministry of Science and Innovation‐Agencia Estatal de Investigación (MCI‐AEI). DG and MSR were supported by the Fundación Científica de la Asociación Española contra el Cáncer (AECC). AFBM and JGM received predoctoral contracts from Foundation La Caixa and the Ministry of Education of Spain (FPI grant BES‐2016‐077901). This work was supported by grants from the European Commission Seventh Framework Programme (ERA‐NET NEURON8‐Full‐815‐094), AEI‐MICIU/FEDER (RTI2018‐095582‐B‐I00 and RED2018‐102723‐T), and the Personalized Medicine and Nanotechnologies in Lung Cancer (iLUNG) and scCANCER programmes from the Comunidad de Madrid (B2017/BMD‐3884 and Y2020/BIO‐6519) to M.M. CNIO is a Severo Ochoa Center of Excellence (AEI‐MICIU CEX2019‐000891‐S)., Maintenance of stemness is tightly linked to cell cycle regulation through protein phosphorylation by cyclin‐dependent kinases (CDKs). However, how this process is reversed during differentiation is unknown. We report here that exit from stemness and differentiation of pluripotent cells along the neural lineage are controlled by CDC14, a CDK‐counteracting phosphatase whose function in mammals remains obscure. Lack of the two CDC14 family members, CDC14A and CDC14B, results in deficient development of the neural system in the mouse and impairs neural differentiation from embryonic stem cells (ESCs). Mechanistically, CDC14 directly dephosphorylates specific proline‐directed Ser/Thr residues of undifferentiated embryonic transcription Factor 1 (UTF1) during the exit from stemness, triggering its proteasome‐dependent degradation. Multiomic single‐cell analysis of transcription and chromatin accessibility in differentiating ESCs suggests that increased UTF1 levels in the absence of CDC14 prevent the proper firing of bivalent promoters required for differentiation. CDC14 phosphatases are dispensable for mitotic exit, suggesting that CDC14 phosphatases have evolved to control stemness rather than cell cycle exit and establish the CDK‐CDC14 axis as a critical molecular switch for linking cell cycle regulation and self‐renewal., Depto. de Bioquímica y Biología Molecular, Fac. de Ciencias Biológicas, TRUE, pub

Objective: The aggressive basal-like molecular subtype of pancreatic ductal adenocarcinoma (PDAC) harbours a ΔNp63 (p40) gene expression signature reminiscent of a basal cell type. Distinct from other epithelia with basal tumours, ΔNp63+ basal cells reportedly do not exist in the normal pancreas. Design: We evaluated ΔNp63 expression in human pancreas, chronic pancreatitis (CP) and PDAC. We further studied in depth the non-cancerous tissue and developed a three-dimensional (3D) imaging protocol (FLIP-IT, Fluorescence Light sheet microscopic Imaging of Paraffin-embedded or Intact Tissue) to study formalin-fixed paraffin-embedded samples at single cell resolution. Pertinent mouse models and HPDE cells were analysed. Results: In normal human pancreas, rare ΔNp63+ cells exist in ducts while their prevalence increases in CP and in a subset of PDAC. In non-cancer tissue, ΔNp63+ cells are atypical KRT19+ duct cells that overall lack SOX9 expression while they do express canonical basal markers and pertain to a niche of cells expressing gastrointestinal stem cell markers. 3D views show that the basal cells anchor on the basal membrane of normal medium to large ducts while in CP they exist in multilayer dome-like structures. In mice, ΔNp63 is not found in adult pancreas nor in selected models of CP or PDAC, but it is induced in organoids from larger Sox9low ducts. In HPDE, ΔNp63 supports a basal cell phenotype at the expense of a classical duct cell differentiation programme. Conclusion: In larger human pancreatic ducts, basal cells exist. ΔNp63 suppresses duct cell identity. These cells may play an important role in pancreatic disease, including PDAC ontogeny, but are not present in mouse models., SCOPUS: ar.j, info:eu-repo/semantics/published

The development of diagnostic tools for measuring a wide spectrum of target analytes, from biomarkers to other biochemical parameters in biological fluids, has experienced a significant growth in the last decades, with a good number of such tools entering the market. Recently, a clear focus has been put on miniaturized wearable devices, which offer powerful capabilities for real-time and continuous analysis of biofluids, mainly sweat, and can be used in athletics, consumer wellness, military, and healthcare applications. Sweat is an attractive biofluid in which different biomarkers could be noninvasively measured to provide rapid information about the physical state of an individual. Wearable devices reported so far often provide discrete (single) measurements of the target analytes, most of them in the form of a yes/no qualitative response. However, quantitative biomarker analysis over certain periods of time is highly demanded for many applications such as the practice of sports or the precise control of the patient status in hospital settings. For this, a feasible combination of fluidic elements and sensor architectures has been sought. In this regard, this paper shows a concise overview of analytical tools based on the use of capillary-driven fluidics taking place on paper or fabric devices integrated with solid-state sensors fabricated by thick film technologies. The main advantages and limitations of the current technologies are pointed out together with the progress towards the development of functional devices. Those approaches reported in the last decade are examined in detail.

Aims/hypothesis: Pancreatic beta cells play a central role in the control of glucose homeostasis by secreting insulin to stimulate glucose uptake by peripheral tissues. Understanding the molecular mechanisms that control beta cell function and plasticity has critical implications for the pathophysiology and therapy of major forms of diabetes. Selective gene inactivation in pancreatic beta cells, using the Cre-lox system, is a powerful approach to assess the role of particular genes in beta cells and their impact on whole body glucose homeostasis. Several Cre recombinase (Cre) deleter mice have been established to allow inactivation of genes in beta cells, but many show non-specific recombination in other cell types, often in the brain. Methods: We describe the generation of Ins1 Cre and Ins1 CreERT2 mice in which the Cre or Cre-oestrogen receptor fusion protein (CreERT2) recombinases have been introduced at the initiation codon of the Ins1 gene. Results: We show that Ins1 Cre mice induce efficient and selective recombination of floxed genes in beta cells from the time of birth, with no recombination in the central nervous system. These mice have normal body weight and glucose homeostasis. Furthermore, we show that tamoxifen treatment of adult Ins1 CreERT2 mice crossed with Rosa26-tdTomato mice induces efficient recombination in beta cells. Conclusions/interpretation: These two strains of deleter mice are useful new resources to investigate the molecular physiology of pancreatic beta cells.

Liquid analysis is key to track conformity with the strict process quality standards of sectors like food, beverage, and chemical manufacturing. In order to analyse product qualities online and at the very point of interest, automated monitoring systems must satisfy strong requirements in terms of miniaturization, energy autonomy, and real time operation. Towards this goal, we present the first implementation of artificial taste running on neuromorphic hardware for continuous edge monitoring applications. We used a solid-state electrochemical microsensor array to acquire multivariate, time-varying chemical measurements, employed temporal filtering to enhance sensor readout dynamics, and deployed a rate-based, deep convolutional spiking neural network to efficiently fuse the electrochemical sensor data. To evaluate performance we created MicroBeTa (Microsensor Beverage Tasting), a new dataset for beverage classification incorporating seven hours of temporal recordings performed over three days, including sensor drifts and sensor replacements. Our implementation of artificial taste is 15× more energy efficient on inference tasks than similar convolutional architectures running on other commercial, low power edge-AI inference devices, achieving over 178× lower latencies than the sampling period of the sensor readout, and high accuracy (97%) on a single Intel Loihi neuromorphic research processor included in a USB stick form factor.

We demonstrate a portable multisensor fusion system for the automated analysis of multiple beverages. The system makes use of compact and low-power-consumption electronic equipment to simultaneously read out an array of microsensors formed by six ion-selective field-effect transistors (ISFETs), one conductivity sensor, one redox potential sensor, and two amperometric microelectrodes. A custom Python application running on a laptop computer receives real-time multivariate data via USB, and provides chemometric models to classify different varieties and to quantify relevant parameters of mineral water and wine. The software also includes a graphical user interface (GUI) to visualize readouts and analytical estimates.