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Colorectal cancer (CRC) is the third most common cancer and the second most frequent cause of cancer-related death worldwide. The detection in plasma samples of autoantibodies against specific tumor-associated antigens has been demonstrated to be useful for the early diagnosis of CRC by liquid biopsy. However, new studies related to the humoral immune response in cancer are needed to enable blood-based diagnosis of the disease. Here, our aim was to characterize the humoral immune response associated with the different p53 and p63 proteoforms derived from alternative splicing and previously described as aberrantly expressed in CRC. Thus, here we investigated the diagnostic ability of the twelve p53 proteoforms and the eight p63 proteoforms described to date, and their specific N-terminal and C-terminal end peptides, by means of luminescence HaloTag beads immunoassays. Full-length proteoforms or specific peptides were cloned as HaloTag fusion proteins and their seroreactivity analyzed using plasma from CRC patients at stages I-IV (n = 31), individuals with premalignant lesions (n = 31), and healthy individuals (n = 48). p53γ, Δ40p53β, Δ40p53γ, Δ133p53γ, Δ160p53γ, TAp63α, TAp63δ, ΔNp63α, and ΔNp63δ, together with the specific C-terminal end α and δ p63 peptides, were found to be more seroreactive against plasma from CRC patients and/or individuals with premalignant lesions than from healthy individuals. In addition, ROC (receiver operating characteristic) curves revealed a high diagnostic ability of those p53 and p63 proteoforms to detect CRC and premalignant individuals (AUC higher than 85%). Finally, electrochemical biosensing platforms were employed in POC-like devices to investigate their usefulness for CRC detection using selected p53 and p63 proteoforms. Our results demonstrate not only the potential of these biosensors for the simultaneous analysis of proteoforms’ seroreactivity, but also their convenience and versatility for the clinical detection of CRC by liquid biopsy. In conclusion, we here show that p53 and p63 proteoforms possess differential seroreactivity in CRC patients in comparison to controls, distinctive from canonical proteins, which should improve the diagnostic panels for obtaining a blood-based biomarker signature for CRC detection.

20 p.-9 fig.-2 tab., Colorectal cancer (CRC) is the second leading cause of cancer-related death worldwide. The 5-year survival rate of CRC patients depends on the stage at diagnosis, being higher than 80% when CRC is diagnosed in the early stages but lower than 10% when CRC is diagnosed in advanced stages. Autoantibodies against specific CRC autoantigens (tumor-associated antigens (TAAs)) in the sera of patients have been widely demonstrated to aid in early diagnosis. Thus, we herein aim to identify autoantigens target of autoantibodies specific to CRC that possess a significant ability to discriminate between CRC patients and healthy individuals by means of liquid biopsy. To that end, we examined the protein content of the exosomes released by five CRC cell lines and tissue samples from CRC patients by means of immunoprecipitation coupled with mass spectrometry analysis. A total of 103 proteins were identified as potential autoantigens specific to CRC. After bioinformatics and meta-analysis, we selected 15 proteins that are more likely to be actual CRC autoantigens in order to evaluate their role in CRC prognosis by Western blot (WB) and immunohistochemistry (IHC). We found dysregulation at the protein level for 11 of these proteins in both tissue and plasma exosome samples from patients, along with an association of nine of these proteins with CRC prognosis. After validation, all but one showed a statistically significant high diagnostic ability to distinguish CRC patients and individuals with premalignant lesions from healthy individuals, either by luminescence Halotag-based beads, or by a multiplexed biosensing platform involving the use of magnetic microcarriers as solid support modified with covalently immobilized Halotag fusion proteins constructed for CRC detection. Taken together, our results highlight the usefulness of the approach defined here to identify the TAAs specific to chronic diseases; they also demonstrate that the measurement of autoantibody levels in plasma against the TAAs identified here could be integrated into a point-of-care (POC) device for CRC detection with high diagnostic ability., This work was supported by the financial support of the PI17CIII/00045 and PI20CIII/00019 grants from the AES-ISCIII program to R.B. The financial support of the PID2019-103899RB-I00 (Ministerio de Ciencia e Innovación) Research Project and the TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (S2018/NMT-4349) to S.C. are gratefully acknowledged. G.D. acknowledges the financial support of PI15/00246 grant of the FIS and Cátedra UAM-Roche en Medicina de Innovación. The FPU predoctoral contract to A.M-C. is supported by the Spanish Ministerio de Educación, Cultura y Deporte. G.S-F. is recipient of a predoctoral contract (1193818N) supported by The Flanders Research Foundation (FWO). M.A-N. was supported by a contract of the Programa Operativo de Empleo Juvenily la Iniciativa de Empleo Juvenil (YEI) with the participation of the Consejería de Educación, Juventud y Deporte de la Comunidad de Madrid y del Fondo Social Europeo. The predoctoral contract from the Spanish Ministerio de Economía y Competitividad (BES-2016-076606, E.P.) and Talento-Contract from Comunidad de Madrid (2019-T2/IND-15965, R.M.T-R.) are also gratefully acknowledged.

Pancreatic ductal adenocarcinoma (PDAC) is the solid tumor with the worst prognosis, representing today the third cause of cancer-related deaths in developed countries and expected to be the second in 2030. Today, CA19-9 remains the only clinically used marker for management of PDAC (FDA-approved as a disease moni- toring marker). This work reports a disposable amperometric immunoplatform for the determination of CA19-9. The immunoplatform skilfully combines the advantages of magnetic microsupports (MBs) for implementation of the immunoassay and amperometric transduction on screen-printed carbon electrodes (SPCEs). The method involves the preparation of sándwich immunocomplexes enzymatically labeled with the enzyme horseradish peroxidase (HRP) on the MBs and uses a detection antibody conjugated to HRP. Once the HRP- labeled sandwich immunocomplexes-bearing MBs were trapped on the SPCE surface, the variation of the catho- dic current was measured in the presence of H2O2 and hydroquinone (HQ), which was directly proportional to the concentration of CA19-9. Under the optimized experimental conditions, the immunoplatform allowed the amperometric determination of CA19-9 standards over the 5.0 to 500 U mL−1 concentration range, with a limit of detection (LOD) value of 1.5 U mL−1 in 1 h. The method exhibits good reproducibility and selectivity and the magnetic immunoconjugates shows a good storage stability. The immunoplatform was applied to the deter- mination of CA19-9 in serum samples of a medium-sized cohort (22 individuals) of healthy subjects and patients diagnosed with PDAC. The obtained results demonstrated the immunoplatform ability to discriminateboth types of individuals within 1 h after sample dilution. The developed immunoplatform represents an improvement in terms of cost, applicability and accessibility compared to the ELISA-based techniques currently used in the clinic.

Magnetic nanoparticles (MNPs) can be used as antibody carriers in a wide range of immunosensing applications. The conjugation chemistry for preparing antibody-MNP bionanohybrids should assure the nanoparticle’s colloidal dispersity, directional conformation and high biofunctionality retention of attached antibodies. In this work, peroxidase (HRP) was selected as model target analyte, and stable antibody-MNP conjugates were prepared using polyaldehyde-dextrans as multivalent linkers, also to prevent nanoparticles agglomeration and steric shielding of non-specific proteins. Under the manipulation of the oxidation variables, MNP-conjugated antibody showed the highest Fab accessibility, of 1.32 μmol analyte per μmol antibody, corresponding to 139 μmol aldehyde per gram of nanocarrier (5 mM NaIO4, 4 h). Demonstrating anti-interference advantage up to 10% serum, colorimetric immunoassay gave a detection limit (LOD) of 300 ng mL− 1 , while electrochemical transduction led to a considerable (680 times) improvement, with a LOD of 0.44 ng mL− 1 . In addition, polyaldehydedextran showed priority over polycarboxylated-dextran as the multivalent antibody crosslinker for MNPs in terms of sensitivity and LOD value, while immunosensors constructed with carboxylated magnetic microbeads (HOOC-MBs) outperformed MNPs-based immunoplatforms. This work sheds light on the importance of surface chemistry (type and density of functional groups) and the dimension (nanosize vs micrometer) of magnetic carriers to conjugate antibodies with better directional orientation and improve the analytical performance of the resulting immunosensors.

Understanding and assessing endocrine response to stress is crucial to human performance analysis, stress-related disorder diagnosis, and mental health monitoring. Current approaches for stress monitoring are largely based on questionnaires, which could be very subjective. To avoid stress-inducing blood sampling and to realize continuous, non-invasive, and real-time stress analysis at the molecular levels, we investigate the dynamics of a stress hormone, cortisol, in human sweat using an integrated wireless sensing device. Highly sensitive, selective, and efficient cortisol sensing is enabled by a flexible sensor array that exploits the exceptional performance of laser-induced graphene for electrochemical sensing. Herein, we report the first cortisol diurnal cycle and the dynamic stress response profile constructed from human sweat. Our pilot study demonstrates a strong empirical correlation between serum and sweat cortisol, revealing exciting opportunities offered by sweat analysis toward non-invasive dynamic stress monitoring via wearable and portable sensing platforms.

The development of versatile and sensitive biotools to quantify specific SARS-CoV-2 immunoglobulins in SARS-CoV-2 infected and non-infected individuals, built on the surface of magnetic microbeads functionalized with nucleocapsid (N) and in-house expressed recombinant spike (S) proteins is reported. Amperometric interrogation of captured N- and S-specific circulating total or individual immunoglobulin (Ig) isotypes (IgG, IgM, and IgA), subsequently labelled with HRP-conjugated secondary antibodies, was performed at disposable single or multiplexed (8×) screen-printed electrodes using the HQ/HRP/H2 O2 system. The obtained results using N and in-house expressed S ectodomains of five SARS-CoV-2 variants of concern (including the latest Delta and Omicron) allow identification of vulnerable populations from those with natural or acquired immunity, monitoring of infection, evaluation of vaccine efficiency, and even identification of the variant responsible for the infection. The financial support of PID2019-103899RB-I00 (Spanish Ministerio de Ciencia e Innovación) Research Project and the TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (Grant S2018/NMT-4349) is grate fully acknowledged. R.B. acknowledges the financial support of the PI20CIII/00019 grant from the AES-ISCIII program. R.M.T.-R. acknowledges Talento-Contract from Comunidad de Madrid (2019-T2/IND-15965). A.M.-C. ac knowledges a FPU predoctoral contract supported by the Spanish Ministerio de Educación, Cultura y Deporte. V.M. acknowledges the financial support of the COV20_00679 grant of the COVID-19 Research Call (ISCIII-Spanish Ministry of Science and Innovation). Sí

Growth arrest-specific 6 (GAS6) protein plays a key role in processes related to proliferation, inflammation, angiogenesis, and atherosclerotic plaque formation. In addition, it has been reported that plasma levels of GAS6 are related to cancer prognosis and other relevant pathologies, such as heart failure or sepsis. We report here the first electrochemical immunoplatform for the determination of GAS6, which has demonstrated to be competitive with other available methodologies in terms of cost, simplicity, and decentralized application. The developed immunoplatform involves a sandwich immunoassay using magnetic microparticles (MBs) and uses amperometric detection at disposable screen-printed carbon electrodes (SPCEs). The MBs were modified with an antibody specific to GAS6 for its selective capture, which is further recognized by a biotinylated secondary antibody subsequently labeled with a streptavidin-horseradish peroxidase (Strep-HRP) conjugate. The electrochemical detection was carried out using the hydroquinone (HQ)/H2O2 system. The developed bioplatform exhibits a great selectivity and low limit of detection (27 pg/mL) that allowed the determination of the GAS6 circulating level in plasma samples from patients suffering heart failure (HF) and diagnosed with pancreatic ductal adenocarcinoma (PDAC), as well as the determination of the target protein in raw secretomes of human colorectal cancer cell lines., This work is part of the POSITION-II project fundedby the ECSEL Joint Undertaking under grant numberEcsel-783132-Position-II-2017-IA;www.position-2.eu,and PCI2018-093067 (Spanish Ministerio de Ciencia eInnovación) to M.P. The financial support of PID2019-103899RB-I00 (Spanish Ministerio de Ciencia e Inno-vación) Research Project to S.C., PI17CIII/00045 andPI20CIII/00019 grants from the AES-ISCIII program toR.B. and the TRANSNANOAVANSENS-CM Program fromthe Comunidad de Madrid (Grant S2018/NMT-4349) toS.C., RTI2018-095672-B-I00 (Spanish Ministerio de Cienciae Innovación) to P.G.F.; Fundació la Marató de TV3 project081010toM.B.; researchprojectPI20/00625,fromtheAES-ISCIII/FEDER program, to P.N, are gratefully acknowl-edged. A. Montero-Calle acknowledges the support of theFPU predoctoral contracts by the Spanish Ministerio deEducación, Cultura y Deporte. G.S-F. is recipient of a pre-doctoral contract (grant number 1193818N) supported byThe Flanders Research Foundation (FWO). C. Muñoz-SanMartín acknowledges a predoctoral contract from Complutense University of Madrid. R.M. Torrente-Rodríguezacknowledges a Talento-Contract from Comunidad deMadrid (2019-T2/IND-15965).

A magnetic beads (MB)-involved amperometric immunosensor for the determination of ST2, a member of the IL1 receptor family, is reported in this work. The method utilizes a sandwich immunoassay and disposable screen-printed carbon electrodes (SPCEs). Magnetic immunoconjugates built on the surface of carboxylic acid-microsized magnetic particles (HOOC-MBs) were used to selectively capture ST2. A biotinylated secondary antibody further conjugated with a streptavidin peroxidase conjugate (Strep-HRP) was used to accomplish the sandwiching of the target protein. The immune platform exhibits great selectivity and a low limit of detection (39.6 pg mL ) for ST2, allowing the determination of soluble ST2 (sST2) in plasma samples from healthy individuals and patients diagnosed with pancreatic ductal adenocarcinoma (PDAC) in only 45 min once the immunoconjugates have been prepared. The good correlation of the obtained results with those provided by an ELISA kit performed using the same immunoreagents demonstrates the potential of the developed strategy for early diagnosis and/or prognosis of the fatal PDAC disease., This research was funded by Spanish Ministerio de Ciencia e Innovación. PID2019- 103899RB-I00, Spanish Ministerio de Ciencia, Innovación y Universidades, RTI2018-095672-B-I00, Comunidad de Madrid, S2018/NMT-4349, and AES-ISCIII/FEDER, PI20/00625

The evolution of human diseases involves a highly dynamic and interactive system of multiple layers of molecular markers. Therefore, it is now fully accepted that simultaneous analysis of molecular markers from the same or different omics layers leads to novel strategies for their realistic early detection and monitoring, thus improving disease prevention, treatment, and outcomes. In this regard, features such as versatility for profiling multiple and/or multi-omics biomarkers, simplicity, affordable cost, markedly shorter analysis time and smaller sample amount required for analyses compared to conventional or state-of-the-art methodologies, make electrochemical bioplatforms suitable alternatives for routine determinations in clinical and basic research settings. The great advances demonstrated by electrochemical biosensors in recent years have gone hand in hand with the development of new electrochemical substrates, attractive surface chemistries, bioassay formats and amplification strategies, but also with the production and application of new bioreceptors, such as those provided by HaloTag and Phage Display technologies, which has allowed these devices to incur and demonstrate pioneering applications. With all this in mind, this communication will discuss bioelectroanalytical tools recently developed by us and our partners potentially transferable to the clinic due to their simplicity, cost, testing time and decentralized character, which have shown pioneering applications to decisively aid in personalized diagnostics by targeting methylation events in nucleic acids both globally and regionally and deregulated autoantibody expression. These biotools have shown to be able to determine simultaneously and with monobasic sensitivity the most common methylated bases in nucleic acids: 5-methylcytosine 5-mC), 5-hydroxymethylcytosine (5-hmC) and N6-methyladenine (6mA) in DNA and N6-methyladenosine (m6A) in RNA, both globally and regionally and without adopting other amplification strategies than conventional enzymatic labeling [1,2]. A multiplexed platform was proposed with application to the global determination of m6A in total RNA extracted from colorectal cancer cells with different metastatic potential, and to the determination of global methylations of 5-mC, 5-hmC and 6mA in genomic DNA extracted from matched tumor and healthy tissues of CRC patients. The obtained results demonstrated the clinical potential of the developed multiplexed bioplatform to discriminate the aggressiveness of tumor cells as well as tumor versus healthy tissues (accuracy, sensitivity, and specificity of 87.5, 100 and 75 %, respectively, using Ward's clustering method) in only 45 min from small amounts (no more than 100 ng) of denatured genomic DNA and crude total RNA without fragmentation and amplification. As for the electroanalytical bioplatforms developed for autoantibodies determination, they have shown ability to identify and confirm the clinical potential of previously undescribed specific autoantibody signatures against i) tumor-associated antigens, identified in the circulation or in tumor exosomes, for early and differential diagnosis of a given neoplasm [3,4]; ii) phage-derived and aberrant HaloTagged peptides to diagnose Alzheimer's disease (AD) [5]; and iii) SARS-CoV-2 serum-specific immunoglobulins to identify vulnerable population or with natural or acquired immunizations, allowing early detection of exaggerated immune system responses or epidemiological studies on the strain responsible for the infection and the efficacy of available vaccines against the different variants of the virus. It is exciting and rewarding to think that we are tackling and developing highly versatile technologies with the potential to efficiently provide relevant information on key aspects of known and unexpected diseases, such as their onset, severity, and immune response, enabling their management in a fast, simple, affordable, and personalized way in diverse settings. [1] Povedano et al. Anal. Chem. 92 (2020) 5604−5612. [2] Povedano et al. Anal. Chim. Acta 1182 (2021) 338946. https://doi.org/10.1016/j.aca.2021.338946. [3] Garranzo-Asensio et al. Theranostics 10(7) (2020) 3022–3034. [4] Montero-Calle et al. Engineering (2021) in press. https://doi.org/10.1016/j.eng.2021.04.026. [5] Valverde et al. Analysis & Sensing 1 (2021) 161-165. The financial support of PID2019-103899RB-I00 (Spanish Ministerio de Ciencia e Innovación), Research Project, PI17CIII/00045 and PI20CIII/00019 grants from the AES-ISCIII program grants from the AES-ISCIII program and the TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (Grant S2018/NMT-4349) are gratefully acknowledged.

Commingled municipal solid waste (MSW) was autoclaved in the presence of saturated steam for 30 min at 145 °C and 600 kPa. The organic fibre fraction from the autoclaved resulting material was examined for biodegradability. Aerobic and anaerobic tests were carried out to characterise the fibre in terms of biodegradation potential, which was moderate (biogas production potential of 251 ± 22 l [biogas] kg⁻¹ [total solids (TS)] and dynamic respiration index (DRI) of 1575 ± 116 mg [O2] kg⁻¹ [TS] h⁻¹). Manual and chemical characterisations were also performed to organic fibre. Following this characterisation, a laboratory-scale thermophilic anaerobic digestion process and a pilot-scale composting process were carried out to determine the possibilities of these biological treatments. In the anaerobic digestion process the biogas yield values obtained were within 0.15-0.21 m³ [biogas] kg⁻¹ [volatile solids (VS)] with an organic loading rate (OLR) of 3 kg [VS] m⁻³ d⁻¹. However, it was difficult to reach the steady state in the anaerobic thermophilic process for the different organic loads tested. Further experiments are necessary to determine the optimal biogas production and performance under these conditions. The composting process performed correctly and the final material was stable (DRI of 504 ± 74 mg [O2] kg⁻¹ [TS] h⁻¹) and with good properties for its application to soil regarding heavy metal contents that corresponding to class B compost, with the exception of some metals that corresponded to class A.

The correlation between onshore and offshore of the volcanic features in a complex volcanic field area is a difficult task, however, it is a fundamental step in order to better understand the geological evolution of such a complex area and for an assessment of geologic hazards. Ischia is a well exposed and densely populated volcanic field located in the Campania volcanic province of Italy. In order to improve our understanding of the recent volcanic history of Ischia Island, high-resolution seismic reflection profiles were used to identify volcanic and sedimentary features in the northern offshore. The volcano stratigraphy interpretation permitted us to recognize seismic units with a reflection-free/chaotic facies. These latter units have been associated with volcanic deposits and correlated to the main volcanic units outcropping on the northern coast of Ischia Island. They are limited in extent and interlayered with eight seismic units with continuous reflectors corresponding to clastic sedimentary units that were deposited during inter-eruptive phases. The main result of this work is the documentation of volcanic activity during the Holocene in the area offshore between Castello d'Ischia, Ischia Porto (mainly effusive products) and Punta della Scrofa (mainly shallow lava domes and dykes). The key volcanic units were mapped and 3D geological models were reconstructed. The reconstruction of the stratigraphic framework offshore a volcanic coast provides a pathway to the investigation of the stratigraphic relationships between inter-eruptive sedimentary deposits and volcanic units, and permits the assessment of a wide and continuous chronostratigraphic framework in a complex area. Furthermore, the onshore-offshore correlation of the main Holocene volcanic units allows us to better estimate their areal distribution, a critical factor in the hazard evaluation of a coastal volcanic area. The application of seismic volcano stratigraphy illustrates the remarkable possibilities that the study of submarine volcanic fields offers.

Altres ajuts: acords transformatius de la UAB Unidad de excelencia María de Maeztu CEX2019-000940-M The absence of sound sampling procedures and statistical analyses to estimate solid waste generation in many developing countries has resulted in incomplete historical records of waste quantity and composition. Data is often arbitrarily aggregated or disaggregated as a function of waste generators to obtain results at the desired spatial level of analysis. Inference fallacies arising from the generalization or individualization of results are almost never considered. In this paper, Panama, one of the fastest-growing developing countries, was used as a case-study to review the main methodological approaches to estimate solid waste generation per capita per day, and at different hierarchical levels (from households to the country). The solid waste generation intensity indicator is used by the Panamanian waste management authority to run the waste management system. It was also the main parameter employed by local and foreign companies to estimate solid waste generation in Panama between 2001 and 2008. The methodological approaches used by these companies were mathematically formalized and classified as per the expressions suggested by Subramanian et al. (2009). Seven inference fallacies (ecological, individualistic, stage, floating population, linear forecasting, average population and mixed spatial levels) were identified and allocated to the studies. Foreign companies committed three of the seven inference fallacies, while one was committed by the local entity. Endogenous knowledge played an important role in these studies to avoid spatial levels mismatch and multilevel measurements appear to produce more reliable information than studies obtained via other means.

The COVID-19 pandemic is an ongoing global challenge for public health systems. Ultrasensitive and early identification of infection is critical in preventing widespread COVID-19 infection by presymptomatic and asymptomatic individuals, especially in the community and in-home settings. We demonstrate a multiplexed, portable, wireless electrochemical platform for ultra-rapid detection of COVID-19: the SARS-CoV-2 RapidPlex. It detects viral antigen nucleocapsid protein, IgM and IgG antibodies, as well as the inflammatory biomarker C-reactive protein, based on our mass-producible laser-engraved graphene electrodes. We demonstrate ultrasensitive, highly selective, and rapid electrochemical detection in the physiologically relevant ranges. We successfully evaluated the applicability of our SARS-CoV-2 RapidPlex platform with COVID-19-positive and COVID-19-negative blood and saliva samples. Based on this pilot study, our multiplexed immunosensor platform may allow for high-frequency at-home testing for COVID-19 telemedicine diagnosis and monitoring., Graphical Abstract, Highlights • Multiplexed detection of SARS-CoV-2 antigen, antibodies, and C-reactive protein • Low-cost and mass manufacturable platform using laser-engraved graphene • Wireless telemedicine sensing platform with rapid sample-to-answer turnaround • Successful device evaluation in both COVID-19 patient blood and saliva samples, Progress and Potential The need for widespread testing to control the spread of COVID-19 has faced challenges due to testing backlogs, limited access to required equipment, and inaccurate assay results. To address this, we propose an ultrasensitive and low-cost telemedicine platform, the SARS-CoV-2 RapidPlex, based on target-specific immunoassays built off laser-engraved graphene for rapid and remote assessment of COVID-19 biomarkers (i.e., nucleocapsid protein, anti-spike protein IgG and IgM, and C-reactive protein). Multiplex sensing of these targets provides information on three key aspects of COVID-19 disease: viral infection, immune response, and disease severity. We successfully demonstrated the platform's applicability using COVID-19-positive and COVID-19-negative serum and saliva samples. The SARS-CoV-2 RapidPlex has the potential to quickly and effectively triage patients and track infection progression, allowing for the clear identification of individuals who are infectious, vulnerable, and/or immune., The SARS-CoV-2 RapidPlex is developed on the basis of laser-engraved graphene immunosensors to electrochemically quantify SARS-CoV-2 nucleocapsid protein, IgG and IgM, and C-reactive protein (CRP). The platform is characterized by rapid detection and high molecular sensitivity and selectivity. Pilot study results demonstrate that it can successfully detect these biomarkers in COVID-19-positive patient serum and saliva samples. CRP results are well correlated with symptom severity, indicating the potential for this integrated system to be used as a diagnostic tool for telemedicine COVID-19 patient care.

Unidad de excelencia María de Maeztu CEX2019-000940-M Developing economies largely rely on imported consumer goods from the manufacturing industries of industrialized economies through free-trade agreements. After consumption, goods end-up in local waste streams and landfilled because of poorly developed waste management systems. This paper proposes a methodology to extend responsibility to exporting country manufacturers for indirect waste disposal in developing countries through imported goods. It establishes a functional relationship between the weight and volume of the imported goods and the local municipal solid waste stream derived from their consumption, by adapting the recycling concepts of by-product and co-product to the municipal solid waste stream derived from the household sector. A lexicon is formalized to conceptualize an extended-producer-responsibility information system operating at the global level between exporting and importing countries. This EPR system i) determines the recyclability, reusability and treatability attributes of imported goods based on their constitutive parts (primary package or product), as well as the material value as per the net value in the global waste market and final destination once consumed, ii) defines specific conditions regarding the goods' materials value and structural configuration of their constitutive parts for inclusion in Free-Trade Agreement clauses, and iii) checks for the fulfilment of these proposed conditions. The proposed methodology was validated with a case study on Panama. It was found that 24%(w/w)-34.5%(v/v) of valued materials derived from goods imported in Panama through FTAs could be exported back to the country of origin, 18%(w/w)-2.8%(v/v) could be locally reused, and 58%(w/w)-62.5%(v/v) locally valorized. Only 16% (w/w)-16%(v/v) would have to be landfilled.

Most of the research works that aim at explaining the relationships between tectonic and volcanism focused a single volcano or eruption. Conversely, this study investigated offshore and onshore areas of the Campania margin in terms of stratigraphy, tectonics, and volcanism at a regional scale. We documented and reconstructed the three-dimensional geometry of several buried volcanoes and volcaniclastic deposits and recognized a complex Late Quaternary tectonic evolution of the region. These results suggest a strict genetic link between rifting and volcanic activity in terms of space-time evolution and because high volumes of magma rose to the surface through regional faults.

Most of the research works that aim at explaining the relationships between tectonic and volcanism focused a single volcano or eruption. Conversely, this study investigated offshore and onshore areas of the Campania margin in terms of stratigraphy, tectonics, and volcanism at a regional scale. We documented and reconstructed the three-dimensional geometry of several buried volcanoes and volcaniclastic deposits and recognized a complex Late Quaternary tectonic evolution of the region. These results suggest a strict genetic link between rifting and volcanic activity in terms of space-time evolution and because high volumes of magma rose to the surface through regional faults.

A number of important drugs used to treat cancer—many of which serve as the backbone of modern chemotherapy regimens—have outdated prescribing information in their drug labeling. The Food and Drug Administration is undertaking a pilot project to develop a process and criteria for updating prescribing information for longstanding oncology drugs, based on the breadth of knowledge the cancer community has accumulated with the use of these drugs over time. This article highlights a number of considerations for labeling updates, including selecting priorities for updating; data sources and evidentiary criteria; as well as the risks, challenges, and opportunities for iterative review to ensure prescribing information for oncology drugs remains relevant to current clinical practice., The U.S. Food and Drug Administration is undertaking a pilot project to develop a process and criteria for updating prescribing information for long‐standing oncology drugs. This article highlights a number of considerations for labeling updates.

This paper describes the first immunosensor reported so far for the highly sensitive determination of the egg white allergen, ovomucoid (OM). The approach involves the selective capture of sandwich immunocomplexes formed by capture antibody-target protein allergen-HRP-labeled detector antibody onto carboxylic acid-functionalized magnetic beads (HOOC-MBs). The resulting magnetic bioconjugates were captured on the surface of disposable screen-printed carbon electrodes (SPCE) to perform amperometric detection at −0.20 V vs. the Ag pseudo-reference electrode in the presence of hydroquinone (HQ) and H2O2. The as prepared electrochemical immunoplatform showed a wide range of linearity (0.3–25 ng mL−1), a LOD of 0.1 ng mL−1, which is much lower than those claimed for commercial ELISA kits, and a great selectivity against other white egg allergenic proteins. The developed immunosensor was successfully employed for the simple and accurate determination of trace amounts of OM in unprocessed (eggs and wheat flour) and baked (bread) food samples.

The paper by De Natale et al. (2016, https://doi.org/10.1002/2015GC006183) reported new stratigraphic and geochronological data for a borehole located at Campi Flegrei volcanic area. The authors failed to mention several significant articles on the geological evolution of the region and proposed an interpretation of their data according to the caldera model. Herein, we provide tectonic and stratigraphic data to support a fault model of ignimbrite emission. This comment will also show the inconsistency of the proposed Campi Flegrei caldera.

This paper describes two different electrochemical affinity biosensing approaches for the simple, fast and bisulfite and PCR-free quantification of 5-methylated cytosines (5-mC) in DNA using the anti-5-mC antibody as biorecognition element. One of the biosensing approaches used the anti-5-mC as capture bioreceptor and a sandwich type immunoassay, while the other one involved the use of a specific DNA probe and the anti-5-mC as a detector bioreceptor of the captured methylated DNA. Both strategies, named for simplicity in the text as immunosensor and DNA sensor, respectively, were implemented on the surface of magnetic microparticles and the transduction was accomplished by amperometry at screen-printed carbon electrodes by means of the hydrogen peroxide/hydroquinone system. The resulting amperometric biosensors demonstrated reproducibility throughout the entire protocol, sensitive determination with no need for using amplification strategies, and competitiveness with the conventional enzyme-linked immunosorbent assay methodology and the few electrochemical biosensors reported so far in terms of simplicity, sensitivity and assay time. The DNA sensor exhibited higher sensitivity and allowed the detection of the gene-specific methylations conversely to the immunosensor, which detected global DNA methylation. In addition, the DNA sensor demonstrated successful applicability for 1 h-analysis of specific methylation in two relevant tumor suppressor genes in spiked biological fluids and in genomic DNA extracted from human glioblastoma cells. The financial support of the Spanish Ministerio de Economía y Competitividad CTQ2015-64402-C2-1-R and SAF2014-53209-R Research Projects, the PI17CIII/00045 research project from AESI and the NANOAVANSENS Program from the Comunidad de Madrid (S2013/MT-3029) and predoctoral contracts from the Spanish Ministerio de Economía y Competitividad (R.M. Torrente-Rodríguez and E. Povedano) and Universidad Complutense de Madrid (V. Ruiz-Valdepeñas Montiel) are also gratefully acknowledged. Sí

A sensitive amperometric immunosensor has been prepared by immobilization of capture antibodies onto gold nanoparticles (AuNPs) grafted on a screen-printed carbon electrode (SPCE) through aryl diazonium salt chemistry using 4-aminothiophenol (AuNPs-S-Phe-SPCE). The immunosensor was designed for the accurate determination of clinically relevant levels of B-type natriuretic peptide (BNP) in human serum samples. The nanostructured electrochemical platform resulted in an ordered layer of AuNPs onto SPCEs which combined the advantages of high conductivity and improved stability of immobilized biomolecules. The resulting disposable immunosensor used a sandwich type immunoassay involving a peroxidase-labeled detector antibody. The amperometric transduction was carried out at −0.20 V (vs the Ag pseudo-reference electrode) upon the addition of hydroquinone (HQ) as electron transfer mediator and H2O2 as the enzyme substrate. The nanostructured immunosensors show a storage stability of at least 25 days, a linear range between 0.014 and 15 ng mL−1, and a LOD of 4 pg mL−1, which is 100 times lower than the established cut-off value for heart failure (HF) diagnosis. The performance of the immunosensor is advantageously compared with that provided with immunosensors prepared by grafting SPCE with p-phenylendiamine (H2N-Phe-SPCE) and attaching AuNPs by immersion into an AuNPs suspension or by electrochemical deposition, as well as with immunosensors constructed using commercial AuNPs-modified SPCEs. The developed immunosensor was applied to the successful analysis of human serum from heart failure (HF) patients upon just a 10-times dilution as sample treatment., The financial support of projects: RetosColaboración RTC-2015- 4184-1 (cofinanced by the Ministry of Economy and Competitivity and FEDER “una manera de hacer Europa”), CTQ2015-70023-R and CTQ2015-64402-C2-1-R (Spanish Ministry of Economy and Competitivity Research Projects) and S2013/MT-3029 (NANOAVANSENS Program from the Comunidad de Madrid) are gratefully acknowledged. R.M. Torrente-Rodríguez acknowledges a predoctoral contract from the Spanish Ministerio de Economía y Competitividad.

Currently, electrochemical nucleic acid-based biosensing methodologies involving hybridization assays, specific recognition of RNA/DNA and RNA/RNA duplexes, and amplification systems provide an attractive alternative to conventional quantification strategies for the routine determination of relevant nucleic acids at different settings. A particularly relevant objective in the development of such nucleic acid biosensors is the design of as many as possible affordable, quick, and simple methods while keeping the required sensitivity. With this aim in mind, this work reports, for the first time, a thorough comparison between 11 methodologies that involve different assay formats and labeling strategies for targeting the same DNA. The assayed approaches use conventional sandwich and competitive hybridization assays, direct hybridization coupled to bioreceptors with affinity for RNA/DNA duplexes, multienzyme labeling bioreagents, and DNA concatamers. All of them have been implemented on the surface of magnetic beads (MBs) and involve amperometric transduction at screen-printed carbon electrodes (SPCEs). The influence of the formed duplex length and of the labeling strategy have also been evaluated. Results demonstrate that these strategies can provide very sensitive methods without the need for using nanomaterials or polymerase chain reaction (PCR). In addition, the sensitivity can be tailored within several orders of magnitude simply by varying the bioassay format, hybrid length or labeling strategy. This comparative study allowed us to conclude that the use of strategies involving longer hybrids, the use of antibodies with specificity for RNA/DNA heteroduplexes and labeling with bacterial antibody binding proteins conjugated with multiple enzyme molecules, provides the best sensitivity.

We report a new concept of a chemiluminescence imaging nanozyme immunoassay (CINIA), in which nanozymes are exploited as catalytic tags for simultaneous multiplex detection of cytokines. The CINIA provides a novel and universal nanozyme-labeled multiplex immunoassay strategy for high-throughput detection of relevant biomarkers and further disease diagnosis.

In the Central Mediterranean the Africa-Eurasia convergence led to the development of complex orogenic systems and backarc basins. Throughout Pliocene-Quaternary times the Southern Apennines accretionary prism migrated towards the Apulia foreland and, contemporaneously, the Tyrrhenian Sea backarc basin opened. In this study, we investigated the offshore of the southern Apulia foreland and the Southern Apennines. Using seismic reflection profiles and well data in a dedicated GIS software, we made a detailed stratigraphic analysis, reconstructed the structural pattern and built 2D geological models of the main geological surfaces. The structure of the Apulia region formed during two main tectonic stages: lower Pleistocene complex extensional faulting developed in a transtensional regime and middle-upper Pleistocene transpression/shortening. The Southern Apennines accretionary prism developed through the Pliocene-early lower Pleistocene and was overprinted by late lower Pleistocene-middle Pleistocene NW-SE sinistral faults. Through linking new data from the accretionary prism and the foreland with previous information from the Tyrrhenian Sea back-arc, we provide an original interpretation for the relationships between the various provinces that make the Central Mediterranean crustal puzzle and reconstruct the main phases of the evolution of its Pliocene-Quaternary orogenic cycle. We recognized collisional stages of uncoupled plates, followed by late lower-middle Pleistocene post-collisional stages of coupled continental plates. This Pleistocene plates re-organization of Central Mediterranean was probably due to the rupture of the Apulia/Ionian slab or to NNW intraplate shortening transmitted from Africa.

Miocene extensional sedimentary basins are being increasingly recognized in Central Mediterranean. However the paleogeographic reconstruction of the Paleo-Tyrrhenian during this time span results a difficult task because the subsequent Pliocene-Quaternary backarc opening moved away these older sedimentary basins. Our study outlines principal deformation events recorded by sediments that were deposited within minor basins of the Tyrrhenian Sea region and provides constraints on the duration of these events. The interpretation of seismic profiles, wells, and outcrops data together with a review of the middle-upper Miocene successions permitted us to recognize Miocene third order depositional sequences, reconstruct the fault pattern, and furnish a detailed evolution of the sedimentary basins that preceded the Pliocene-Quaternary Tyrrhenian high stretching. Based on the stratigraphic and tectonic constraints together with the age migration of the depocenters, we reconstructed a detailed paleogeographic evolution of central Mediterranean. Sedimentary basins architecture and fault pattern indicate a system of approximately N-S oriented normal faults and approximately E-W transform faults that were active since the Late Oligocene. Two period of extensional/transtensional tectonics (late Oligocene-Lower Burdigalian and Upper Langhian-Tortonian) were interrupted by a compressional event (late Burdigalian-Lower Langhian). Tacking in account the position of the Miocene volcanic arc (Sardinia), we propose that the coeval sedimentary basins developed in the central Mediterranean as forearc extensional/transtensional basins during the progressive collision with the African plate and before the Tyrrhenian backarc opening. Unlike published late Oligocene-Tortonian reconstructions of the western-central Mediterranean realm that report a linear migration of backarc extension, our results support a geodynamic model characterized by a progressive deformation of the forearc extension in the overriding plate.

Early and reliable diagnostic of cancer is mandatory to increase patient survival, thus requiring efficient and reliable analytical methods for such a purpose. Within this context, different strategies implying the development of electrochemical biosensors for the sensitive, selective and rapid multiplexed biosensing of genetic or protein cancer-related biomarkers are addressed in this presentation. In particular, novel sensing platforms have been developed for the determination of miRs, interleukin (IL)-8 mRNA, IL-8 protein, and cancer specific receptors. The developed methodologies allow for the determination of the target analytes at clinically relevant levels in complex samples: cancer cells, human tissues cell lysates, serum and raw saliva and can be easily extended to the determination of other relevant biomarkers.

Alpha-lactalbumin (α-LA) is one of the whey proteins in cows' milk that has been identified as allergenic. In this work, we present, for the first time, a very sensitive magnetic beads (MBs)-based immunosensor for the determination of α-LA. A sandwich configuration involving selective capture and horseradish peroxidase-labeled detector antibodies was implemented on carboxylic acid-modified magnetic beads, captured magnetically under the surface of a disposable screen-printed carbon electrode for amperometric detection using the hydroquinone (HQ)/H2O2 system. The α-LA immunosensor exhibited a wide linear range (37.0-5000pg/ml), a low limit of detection (LOD, 11.0pg/ml) and noteworthy selectivity against other non-target proteins. The MBs-based immunosensing platform was applied successfully for the determination of α-LA in several varieties of milk (raw and UHT cows' milk as well as human milk) and infant formulations. The results were corroborated with those obtained using a commercial ELISA method, thereby substantiating the analytical merits of this unique method.

Unidad de excelencia María de Maeztu CEX2019-000940-M Existing studies have studied influencing factors of MSW generation behaviour at different spatial levels of organization, but always one at a time and not simultaneously. Income is a strong influencing factor, affecting MSW generation from the individual to the country level, capable of hiding the effects of the others. This study shows that when MSW generation behaviour is holistically analysed across multiple levels of organization (individuals, households, and communities) hierarchically organized as functional units of MSW generation within a specific study area, it is possible to identify influencing factors in addition to income (education, demographic, health, ethnic, economic activity and financial types) as explanatory variables. Increasing the number of influencing factors of MSW generation makes it possible to create a robust knowledge base for MSW management policies in fast-growing urban areas of developing countries, improving the information used to select proper policies and plans within their MSW management systems and avoiding overlapping policies causing legal gaps. Betania, an urban area of the Panama City district, has been chosen as a case study area. The results show that the household income explains 86% of its memberś MSW generation and the community indigenous population explains 21% of householdś MSW generation. It is concluded that MSW generation is not linear across levels, it has as many degrees of freedom as influencing factors shaping the levels of organization where functional units generating waste exist. Influencing factors appearing at each spatial level affects MSW generation in an interdependent manner in variable degrees of magnitude.

Unidad de excelencia María de Maeztu MdM-2015-0552 This paper studies landfill emissions and the related environmental and health risks in Panama City, with the aim to sensitize the population about the harmful effects of irresponsible resource consumption and non-deliberate solid waste generation that it is disposed of in an uncontrolled manner in landfills. Empirical data on Cerro Patacón, Panama City's landfill was obtained to describe the status of municipal waste disposal. Ten known methane generation models were used to estimate the yearly emission rate of methane from the landfill for a 100-year period starting from its inception in 1986. From the models used, the GasSIM model was chosen to estimate emission rates of six long-term hazardous air pollutants. The AERMOD source dispersion model was used to simulate their atmospheric downwind dispersion by levels of concentration over nearby affected communities; results were mapped in Google Earth. The relative contributions by population of the 32 towns making up Panama City to the forecasted waste generation in 2022 and related hazardous air pollutants emission rates from the landfill were assessed. It was found that Cerro Patacón will generate 45% of the countrywide methane generation by 2022; an average of 47 Gg. The solid waste generated by the 1.5 million inhabitants of Panama City impacts the health of ~73,600 inhabitants in nearby communities through the dispersion of hazardous atmospheric pollutants derived from the landfill. The highest emission rates were from hydrogen sulfide and dichloromethane, which can be largely attributed to the waste generated by the communities of Juan Diaz and Tocúmen. The concentration of hydrogen sulfide and benzene was over the reference concentration (uncertainty factor spanning three orders of magnitude) for all communities and years simulated. The concentration of vinyl chloride was over the RfC for all communities and years simulated, except in 2018 for 12 communities.

13 p.-5 fig-1 tab., Phenotypic drug discovery must take advantage of the large amount of clinical data currently available. In this sense, the impact of microRNAs (miRs) on human disease and clinical therapeutic responses is becoming increasingly well documented. Accordingly, it might be possible to use miR-based signatures as phenotypic read-outs of pathological status, for example in cancer. Here, we propose to use the information accumulating regarding the biology of miRs from clinical research in the preclinical arena, adapting it to the use of miR biosensors in the earliest steps of drug screening. Thus, we have used an amperometric dual magnetosensor capable of monitoring a miR-21/miR-205 signature to screen for new drugs that restore these miRs to non-tumorigenic levels in cell models of breast cancer and glioblastoma. In this way we have been able to identify a new chemical entity, 11PS04 ((3aR,7aS)-2-(3-propoxyphenyl)-7,7a-dihydro-3aH-pyrano[3,4-d]oxazol-6(4H)-one), the therapeutic potential of which was suggested in mechanistic assays of disease models, including 3D cell culture (oncospheres) and xenografts. These assays highlighted the potential of this compound to attack cancer stem cells, reducing the growth of breast and glioblastoma tumors in vivo. These data demonstrate the enhanced chain of translatability of this strategy, opening up new perspectives for drug-discovery pipelines and highlighting the potential of miR-based electro-analytical sensors as efficient tools in modern drug discovery., This work was supported by grants CTQ2014-52213-R, CTQ2015-64402-C2-1-R and CTQ2015-64402-C2-2-R (Spanish Ministry of Economy and Competitiveness, MINECO), CM S2011/BMED-2353 (Comunidad de Madrid), and FEDER Funds.

The Tyrrhenian Sea (Central Mediterranean) is an extensional area that formed during the collision between Africa and Eurasia. Through the analysis of well logs and seismic profiles we reconstructed the stratigraphic successions and the structural style of the Tyrrhenian sedimentary basins. The sequence stratigraphy interpretation of well logs permitted us to recognize third and fourth order depositional sequences and the stratigraphic signatures of the rift stages. The study area features several sediment overfilled/balanced infill basins that resulted from the superposition of several rifting events characterized by high rates of basin subsidence. These outcomes led us to propose five rifting episodes occurred from Miocene to Quaternary affecting the upper plate. The suggested geodynamic scenario takes into account a progressive rupture of the slab and formation of sedimentary basins in the upper plate.

Coeval rifting processes affecting both backarc, thrust belt and foreland occurred in offshore western Sicily (Central Mediterranean) as indicated by the analysis of a large seismic data set calibrated with well data, onshore data and dredges in a dedicated GIS software environment. Structural pattern of the investigated area, 3-D geological models and a Pliocene-Lower Pleistocene evolution of extensional sedimentary basins have been reconstructed. In detail, two zones characterized by basins with different directions of extension have been recognized: one trending NE-SW, located south of the Egadi Islands; another trending E-W, positioned north of the Egadi Islands. The first group of basins represents the continuation of those of the Africa foreland basin (Pelagian Block), as the second one characterizes the opening of the backarc (southern Tyrrhenian margin). Both basins affect the collisional chain (Maghrebian thrust belt). An eastward progression of extension, from lower Pliocene to lower Pleistocene, based on different onset of the basin fill, has been documented in the Southern Tyrrhenian margin. The study area represents the meeting place of backarc and foreland rifting that affected the late Miocene thrust belt. The recognized pattern of extensional deformation of the upper plate adds some constraint on the geodynamic evolution of the African and Eurasia plates and supports a model of westward dipping slab in the Central Mediterranean. It is hypothesized that the along-trench variation of the nature of the plates, due to paleo-margin configuration, has a profound impact on the upper crustal deformation during the convergence and thus the meeting place of backarc and foreland rifting could represent the surficial signature of the slab detachment since Pliocene times.

This work describes the preparation of an immunoplatform for the sensitive and selective determination of N6-methyladenosine (m6A). The simple and fast protocol involves for the first time the use of micromagnetic immunoconjugates to establish a direct competitive assay between the m6A target and a biotinylated RNA oligomer bearing a single m6A enzymatically labelled with a commercial conjugate of streptavidin-peroxidase (Strep-HRP) as tracer. The cathodic current change measured in the presence of H2O2/hydroquinone (HQ) at screen-printed carbon electrodes (SPCEs) upon surface capturing the magnetic bioconjugates is inversely proportional to the m6A target concentration. After evaluating the effect of key variables, the analytical characteristics were established for the determination of three different targets: the N6-methyladenosine-5′-triphosphate (m6ATP) ribonucleotide, a short synthetic RNA oligomer bearing a single m6A and the positive control provided in a commercial colorimetric kit for m6A-RNA quantification. The obtained results show that this immunoplatform is competitive with other methods reported to date, achieving an improved sensitivity (limit of detection of 0.9 pM for the short synthetic oligomer) using a much simpler and faster protocol (~1 h) and disposable electrodes for the transduction. Furthermore, the applicability for discriminating the metastatic potential of cancer cells by directly analyzing a small amount of raw total RNA without enriching or fragmenting was also preliminary assessed.