Search

Abstract Background The physiological relevance of cell-to-cell communication mediated by small extracellular vesicle-encapsulated microRNAs (sEV-miRNAs) remains debated because of the limiting representativity of specific miRNAs within the extracellular pool. We hypothesize that sEV-miRNA non-canonical function consisting of the stimulation of Toll-like receptor 7 (TLR7) may rely on a global shift of the sEV cargo rather than on the induction of one or few specific miRNAs. Psoriasis represents an ideal model to test such hypothesis as it is driven by overt activation of TLR7-expressing plasmacytoid dendritic cells (pDCs) following keratinocyte damage. Methods To mimic the onset of psoriasis, keratinocytes were treated with a cocktail of psoriatic cytokines or UV-irradiated. SmallRNA sequencing was performed on sEVs released by healthy and UV-treated keratinocytes. sEV-miRNAs were analyzed for nucleotide composition as well as for the presence of putative TLR7-binding triplets. Primary human pDCs where stimulated with sEVs +/- inhibitors of TLR7 (Enpatoran), of sEV release (GW4869 + manumycin) and of TLR7-mediated pDC activation (anti-BDCA-2 antibody). Secretion of type I IFNs and activation of CD8+T cells were used as readouts. qPCR on psoriatic and healthy skin biopsies was conducted to identify induced miRNAs. Results sEV-miRNAs released by damaged keratinocytes revealed a significantly higher content of TLR7-activating sequences than healthy cells. As expected, differential expression analysis confirmed the presence of miRNAs upregulated in psoriatic skin, including miR203a. More importantly, 76.5% of induced miRNAs possessed TLR7-binding features and among these we could detect several previously demonstrated TLR7 ligands. In accordance with this in silico analysis, sEVs from damaged keratinocytes recapitulated key events of psoriatic pathogenesis by triggering pDCs to release type I interferon and activate cytotoxic CD8+T cells in a TLR7- and sEV-dependent manner. Discussion Our results demonstrate that miR203a is just one paradigmatic TLR7-activating miRNA among the hundreds released by UV-irradiated keratinocytes, which altogether trigger pDC activation in psoriatic conditions. This represents the first evidence that cell damage shifts the miRNA content of sEVs towards a TLR7-activating cargo capable to propagate inflammation and immunity, offering strong support to the physiological role of systemic miRNA-based cell-to-cell communication.

Abstract Extracellular vesicles (EVs) are nanosized heat-stable vesicles released by virtually all cells in the body, including tumor cells and tumor-infiltrating dendritic cells (DCs). By carrying molecules from originating cells, EVs work as cell-to-cell communicators in both homeostasis and cancer but may also represent valuable therapeutic and diagnostic tools. This review focuses on the role of tumor-derived EVs (TEVs) in the modulation of DC functions and on the therapeutic potential of both tumor- and DC-derived EVs in the context of immunotherapy and DC-based vaccine design. TEVs were originally characterized for their capability to transfer tumor antigens to DCs but are currently regarded as mainly immunosuppressive because of the expression of DC-inhibiting molecules such as PD-L1, HLA-G, PGE2 and others. However, TEVs may still represent a privileged system to deliver antigenic material to DCs upon appropriate engineering to reduce their immunosuppressive cargo or increase immunogenicity. DC-derived EVs are more promising than tumor-derived EVs since they expose antigen-loaded MHC, costimulatory molecules and NK cell-activating ligands in the absence of an immunosuppressive cargo. Moreover, DC-derived EVs possess several advantages as compared to cell-based drugs such as a higher antigen/MHC concentration and ease of manipulation and a lower sensitivity to immunosuppressive microenvironments. Preclinical models showed that DC-derived EVs efficiently activate tumor-specific NK and T cell responses either directly or indirectly by transferring antigens to tumor-infiltrating DCs. By contrast, however, phase I and II trials showed a limited clinical efficacy of EV-based anticancer vaccines. We discuss that the future of EV-based therapy depends on our capability to overcome major challenges such as a still incomplete understanding of their biology and pharmacokinetic and the lack of standardized methods for high-throughput isolation and purification. Despite this, EVs remain in the limelight as candidates for cancer immunotherapy which may outmatch cell-based strategies in the fullness of their time.

IntroductionTissue engineering has advanced significantly in recent years, owing primarily to additive manufacturing technology and the combination of biomaterials and cells known as 3D cell printing or Bioprinting. Nonetheless, various obstacles remain developing adequate 3D printed structures for biomedical applications, including bioinks optimization to meet biocompatibility and printability standards. Hydrogels are among the most intriguing bioinks because they mimic the natural extracellular matrix found in connective tissues and can create a highly hydrated environment that promotes cell attachment and proliferation; however, their mechanical properties are weak and difficult to control, making it difficult to print a proper 3D structure.MethodsIn this research, hydrogels based on Alginate and Gelatin are tested to evaluate the metabolic activity, going beyond the qualitative evaluation of cell viability. The easy-to-make hydrogel has been chosen due to the osmotic requirements of the cells for their metabolism, and the possibility to combine temperature and chemical crosslinking. Different compositions (%w/v) are tested (8% gel-7% alg, 4% gel-4% alg, 4% gel-2% alg), in order to obtain a 3D structure up to 10.3 ± 1.4 mm.ResultsThe goal of this paper is to validate the obtained cell-laden 3D structures in terms of cell metabolic activity up to 7 days, further highlighting the difference between printed and not printed cell-laden hydrogels. To this end, MS5 cells viability is determined by implementing the live/dead staining with the analysis of the cellular metabolic activity through ATP assay, enhancing the evaluation of the actual cells activity over cells number.DiscussionThe results of the two tests are not always comparable, indicating that they are not interchangeable but provide complementary pieces of information.

This article aims to reflect on the images of childhood in videos featuring child YouTubers playing, analyzing the nature of play portrayed in them and its relationship with the child’s right to play and be protected against any form of exploitation. Method: A documentary study of 100 videos shared on YouTube was conducted, subjected to categorical content analysis with an emphasis on the modes of participation of adults and children in dialogues with the platform’s languages present in the videos. Results and Discussion: The boundaries between children’s artistic expression and child labor exploitation are becoming increasingly challenging, with legal discussions presenting difficulties in regulation due to the home environment and predominant parental control. The analysis reveals how the videos are perceived as standardized commodities, hiding the concrete work behind an image of apparent spontaneity. Conclusion: The research highlights contractual digital risks for children, focusing on those related to commercialization. The analyzed videos reflect an instrumentalization of the child’s basic right to play, associated with market interests, an aspect that takes on the contours of child labor exploitation, compromising the freedom to play spontaneously.

Objectives: Meningitis is a medical emergency, and it is crucial to diagnose it accurately and promptly in order to manage patients effectively. It would, therefore, be essential to introduce and have fast, accurate, and user-friendly methods to determine the cause of these infections. This study aimed to demonstrate a potentially cost-effective new approach for detecting meningitis using a paper-based vertical flow microarray, which could be useful in settings with limited resources. Methods: We describe a multiplex paper microarray for detecting Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, and Salmonella spp. by the passive vertical flow of PCR-amplified clinical samples. A multibiotinylated amplicon was obtained as a product of PCR in the presence of both a biotinylated primer and biotin-11-dUTP. An enhancement step based on an enzyme-free gold enhancement protocol was also used to facilitate visual detection. Results: This study showed that the vertical flow microarray (previously evaluated for one pathogen) can discriminately detect the amplification results down to the 102 copies of DNA limit for four meningitis pathogens in a multiplexed set-up. The study further demonstrated the ability of this device and setup to detect three of the four pathogens from clinical biosamples. Discussion: This study demonstrated the capacity of a vertical flow microarray device to detect amplification products for four prevalent meningitis pathogens in a multiplex format. The vertical flow microarray demonstrated consistent visualization of the expected gene amplification results; however, indicating limitations in the pre- and amplification steps. This study highlights the potential of this multiplexing method for diagnosing meningitis and other syndromic diseases caused by various pathogens, especially in resource-limited areas.

IntroductionLung carcinoids (LCs) are a type of neuroendocrine tumor (NET) that originate in the bronchopulmonary tract. LCs account for 20–25% of all NETs and approximately 1–2% of lung cancers. Given the highly vascularized nature of NETs and their tendency to overexpress vascular growth factor receptors (VEGFR), inhibiting angiogenesis appears as a potential therapeutic target in slowing down tumor growth and spread. This study evaluated the long-term antitumor activity and related mechanisms of axitinib (AXI), a VEGFR-targeting drug, in LC cell lines.MethodsThree LC cell lines (NCI-H727, UMC-11 and NCI-H835) were incubated with their respective EC50 AXI concentrations for 6 days. At the end of the incubation, FACS experiments and Western blot analyses were performed to examine changes in the cell cycle and the activation of apoptosis. Microscopy analyses were added to describe the mechanisms of senescence and mitotic catastrophe when present.ResultsThe primary effect of AXI on LC cell lines is to arrest tumor growth through an indirect DNA damage. Notably, AXI triggers this response in diverse manners among the cell lines, such as inducing senescence or mitotic catastrophe. The drug seems to lose its efficacy when the DNA damage is mitigated, as observed in NCI-H835 cells.ConclusionThe ability of AXI to affect cell viability and proliferation in LC tumor cells highlights its potential as a therapeutic agent. The role of DNA damage and the consequent activation of senescence seem to be a prerequisite for AXI to exert its function.

Our study was designed to develop a customisable, wearable, and comfortable medical device – the text so-called “MyPAD” that monitors the fullness of the bladder, triggering an alarm indicating the need to void, in order to prevent badwetting – i.e., treating Nocturnal Enuresis (NE) at the text pre-void stage using miniaturised mechatronics with Artificial Intelligence (AI). The developed features include: multiple bespoke ultrasound (US) probes for sensing, a bespoke electronic device housing custom US electronics for signal processing, a bedside alarm box for processing the echoed pulses and generating alarms, and a phantom to mimic the human body. The validation of the system is conducted on the text tissue-mimicking phantom and volunteers using Bidirectional Long Short-Term Memory Recurrent Neural Networks (Bi-LSTM-RNN) and Reinforcement Learning (RL). A Se value of 99% and a Sp value of 99.5% with an overall accuracy rate of 99.3% are observed. The obtained results demonstrate successful empirical evidence for the viability of the device, both in monitoring bladder expansion to determine voiding need and in reinforcing the continuous learning and customisation of the device for bladder control through consecutive uses. Clinical impact: MyPAD will treat the NE better and efficiently against other techniques currently used (e.g., post-void alarms) and will i) replace those techniques quickly considering sufferers’ condition while being treated by other approaches, and ii) enable children to gain control of incontinence over time and consistently have dry nights. Category: Early/Pre-Clinical Research

The use of alternative renewable sources to totally or partially replace bitumen is one of the most current challenges in the road pavement sector. The growing energy cost and environmental concerns lead to the necessity to find alternative solutions, by supporting at the same time sustainability and circular economy principles. Within this framework, this paper presents the application of a powder lignin, a natural bio-polymer deriving from by-products of wood pulp and paper industry, to replace part of bitumen. The bituminous blend consisting in 70% of bitumen and 30% of powder lignin (by weight) was made in laboratory through the use of a high shear stirring mixer, and a reference plain bitumen characterized by a similar consistency (i.e., same penetration grade) was used as comparison. Then, an extensive investigation on chemical and rheological properties of the bio-binder is presented. Fourier transform infrared spectroscopy (FTIR), saturates, aromatics, resins and asphaltenes (SARA), bending beam rheometer (BBR), frequency sweep tests and multiple stress creep recovery tests (MSCR) with a dynamic shear rheometer (DSR) were performed. Moreover, unaged, short- and long-term aging conditions were considered. Results indicated that powder lignin dominates the rheological behavior of the bio-binder and, from chemical analysis, it seems that it partially acts as a filler and partially as a binder. This would result in improved performances at both low and high temperatures, leading to a wider temperature range of performance grade (PG). Moreover, despite a stiffening effect is recognized, lignin also offers an antioxidant potentiality, reducing the aging susceptibility of the investigated bio-binder.

BackgroundDuring its life cycle, the human pathogen Trypanosoma cruzi must quickly adapt to different environments, in which the variation in the gene expression of the regulatory U-rich RNA-binding protein 1 (TcUBP1) plays a crucial role. We have previously demonstrated that the overexpression of TcUBP1 in insect-dwelling epimastigotes orchestrates an RNA regulon to promote differentiation to infective forms.MethodsIn an attempt to generate TcUBP1 knockout parasites by using CRISPR-Cas9 technology, in the present study, we obtained a variant transcript that encodes a protein with 95% overall identity and a modified N-terminal sequence. The expression of this mutant protein, named TcUBP1mut, was notably reduced compared to that of the endogenous form found in normal cells. TcUBP1mut-knockdown epimastigotes exhibited normal growth and differentiation into infective metacyclic trypomastigotes and were capable of infecting mammalian cells.ResultsWe analyzed the RNA-Seq expression profiles of these parasites and identified 276 up- and 426 downregulated genes with respect to the wildtype control sample. RNA-Seq comparison across distinct developmental stages revealed that the transcriptomic profile of these TcUBP1mut-knockdown epimastigotes significantly differs not only from that of epimastigotes in the stationary phase but also from the gene expression landscape characteristic of infective forms. This is both contrary to and consistent with the results of our recent study involving TcUBP1-overexpressing cells.ConclusionTogether, our findings demonstrate that the genes exhibiting opposite changes under overexpression and knockdown conditions unveil key mRNA targets regulated by TcUBP1. These mostly encompass transcripts that encode for trypomastigote-specific surface glycoproteins and ribosomal proteins, supporting a role for TcUBP1 in determining the molecular characteristics of the infective stage.

Previous studies have shown that overexpression of the Trypanosoma cruzi U-rich RNA-binding protein 1 (TcUBP1) in insect-dwelling epimastigotes results in a gene expression pattern resembling that of the infective form of the pathogen. Here, we used CRISPR-Cas9-induced edition of TcUBP1 and full-length protein overexpression in epimastigote cells to monitor transcriptomic changes during the epimastigote-to-metacyclic trypomastigote stage transition of T. cruzi. This dataset includes the bioinformatics analysis of three different RNA-seq samples, each with three biological replicates, showing differential mRNA abundances. The current transcriptome report has the potential to shed light on the quantitative variances in the expression of significantup- or down-regulated mRNAs as a consequence of the levels of the UBP1 protein. Raw data files were deposited at the NCBI Sequence Read Archive - SRA at http://ncbi.nlm.nih.gov/Traces/sra/sra.cgi with accession numbers PRJNA907231 and PRJNA949967.

We present data on the proteome of the Trypanosoma cruzi epimastigote cells overexpressing the U-rich RNA-binding protein 1 (UBP1). The role of this regulatory protein during the epimastigote-to-metacyclic trypomastigote stage transition was clearly established by our group at the transcriptome level; nevertheless, the impact of UBP1 overexpression on protein synthesis is not known. To address this question, we performed shotgun label-free quantification proteomics using an in vitro system based on the tetracycline-inducible expression of TcUBP1 and epimastigote wildtype cells. Using tryptic peptide digestion and LC-MS/MS analysis with Orbitrap technology, this data file describes the proteome of three biological samples per condition and yields 1637 correctly quantified proteins. The statistical comparisons of the two analyzed groups within the Proteome Discoverer platform identified 379 differentially expressed proteins, with 207 being up-regulated and 172 being down-regulated. In addition, profile plots and heatmap analysis to visualize the distribution of protein abundances within replicates are also presented. Data are available via ProteomeXchange with identifier PXD047761.

This study investigated the impact of early prosthetic intervention on children with upper limb agenesis or amputation. The aim was to assess both how early prosthetics improve a child’s movement and coordination skills, as well as how satisfied both the child and their parents are with the prosthesis. Twenty-nine children were evaluated using the Unilateral Below Elbow Test (UBET) and Child Amputee Prosthetics Project—Prothesis Satisfaction Inventory (CAPP-PSI). Results suggest that while children performed tasks faster and more fluidly without a prosthesis, they were satisfied with the prostheses and the service provided. This may be due to ongoing adaptation to the new device. The single child re-evaluated at 12 months showed improvement in prosthesis use, highlighting the potential benefits of early intervention coupled with motor re-education. Further research is needed to optimize prosthetic features and address initial challenges associated with prosthesis use.

Congenital and chronic liver diseases have a substantial health burden worldwide. The most effective treatment available for these patients is whole organ transplantation; however, due to the severely limited supply of donor livers and the side effects associated with the immunosuppressive regimen required to accept allograft, the mortality rate in patients with end-stage liver disease is annually rising. Stem cell-based therapy aims to provide alternative treatments by either cell transplantation or bioengineered construct transplantation. Human amnion epithelial cells (AEC) are a widely available, ethically neutral source of cells with the plasticity and potential of multipotent stem cells and immunomodulatory properties of perinatal cells. AEC have been proven to be able to achieve functional improvement towards hepatocyte-like cells, capable of rescuing animals with metabolic disorders; however, they showed limited metabolic activities in vitro. Decellularised extracellular matrix (ECM) scaffolds have gained recognition as adjunct biological support. Decellularised scaffolds maintain native ECM components and the 3D architecture instrumental of the organ, necessary to support cells’ maturation and function. We combined ECM-scaffold technology with primary human AEC, which we demonstrated being equipped with essential ECM-adhesion proteins, and evaluated the effects on AEC differentiation into functional hepatocyte-like cells (HLC). This novel approach included the use of a custom 4D bioreactor to provide constant oxygenation and media perfusion to cells in 3D cultures over time. We successfully generated HLC positive for hepatic markers such as ALB, CYP3A4 and CK18. AEC-derived HLC displayed early signs of hepatocyte phenotype, secreted albumin and urea, and expressed Phase-1 and -2 enzymes. The combination of liver-specific ECM and bioreactor provides a system able to aid differentiation into HLC, indicating that the innovative perfusion ECM-scaffold technology may support the functional improvement of multipotent and pluripotent stem cells, with important repercussions in the bioengineering of constructs for transplantation.

The 2030 Global Task Force on Cholera Control Roadmap hinges on strengthening the implementation of multistranded cholera interventions, including community engagement and health system strengthening. However, a composite picture of specific facilitators and barriers for these interventions and any overlapping factors existing between the two, is lacking. Therefore, this study aims to address this shortcoming, focusing on cholera-reporting countries, which are disproportionately affected by cholera and may be cholera endemic. A scoping methodology was chosen to allow for iterative mapping, synthesis of the available research and to pinpoint research activity for global and local cholera policy-makers and shareholders. Using the Arksey and O’Malley framework for scoping reviews, we searched PubMed, Web of Science and CINAHL. Inclusion criteria included publication in English between 1990 and 2021 and cholera as the primary document focus in an epidemic or endemic setting. Data charting was completed through narrative descriptive and thematic analysis. Forty-four documents were included, with half relating to sub-Saharan African countries, 68% (30/44) to cholera endemic settings and 21% (9/44) to insecure settings. We identified four themes of facilitators and barriers to health systems strengthening: health system cooperation and agreement with external actors; maintaining functional capacity in the face of change; good governance, focused political will and sociopolitical influences on the cholera response and insecurity and targeted destruction. Community engagement had two themes: trust building in the health system and growing social cohesion. Insecurity and the community; cooperation and agreement; and sociopolitical influences on trust building were themes of factors acting at the interface between community engagement and health system. Given the decisive role of the community–health system interface for both sustained health system strengthening and community engagement, there is a need to advocate for conflict resolution, trust building and good governance for long-term cholera prevention and control in cholera reporting countries.

Paolo Vincenzi,1 Massimo Stronati,2 Paolo Garelli,3 Diletta Gaudenzi,4 Gianfranco Boccoli,3 Roberto Starnari2 1Department of General Surgery, Ospedali Riuniti Marche Nord, Pesaro, Italy; 2Department of Anesthesiology, IRCSS-INRCA, Ancona, Italy; 3Department of General Surgery, IRCSS-INRCA, Ancona, Italy; 4Department of Perioperative Services, AOU “Ospedali Riuniti di Ancona”, Ancona, ItalyCorrespondence: Paolo Vincenzi, Department of General Surgery, Ospedali Riuniti Marche Nord, Piazzale Cinelli n 1, Pesaro, 61121, Italy, Tel +39 3394578495, Email paolo.vincenzi1981@gmail.comPurpose: Several studies have applied lumbar spinal anesthesia (SA) with isobaric/hyperbaric bupivacaine and opioids in elective laparoscopic cholecystectomy (LC), documenting a superiority of the methodic over general anesthesia (GA) in terms of perioperative pain, nausea, and vomiting, though with a notable incidence of intraoperative right shoulder pain, potentially responsible for conversion to GA. This case series presents an opioid-free scheme of segmental thoracic spinal anesthesia (STSA) with hypobaric ropivacaine, reporting its benefits mainly in terms of shoulder pain occurrence.Patients and Methods: Hypobaric STSA was performed in nine patients undergoing elective LC between May 1 and September 1, 2022. The level of the needle insertion was included between T8 and T9, via a median or a paramedian approach. Midazolam (0.03 mg/kg) and Ketamine (0.3 mg/kg) were used as adjuvants for intrathecal sedation, followed by the administration of hypobaric ropivacaine 0.25% at a dose of 5 mg and then isobaric ropivacaine at a dose of 10 mg. Patients were placed in anti-Trendelenburg position for the entire duration of surgery. LC was conducted through the standard 3 or 4 ports technique with pneumoperitoneum maintained at a pressure of 8– 10 mmHg.Results: Mean patient age was 75.7 (± 17.5) years, with a mean ASA score and Charlson comorbidity index (CCI) of 2.7 (± 0.7) and 4.9 (± 2.7), respectively. STSA was completed without complications in all patients, with no need for conversion to GA. Mean operative time and SA duration were 37.5 (± 8.7) and 145.2 (± 21.8) min, respectively. Intraoperatively, no shoulder or abdominal pain and nausea were reported, with only four and two patients requiring vasopressor and sedative intravenous drugs, respectively. Postoperatively, overall mean VAS pain score and within the first 12 hafter surgery were 3 (± 2) and 4 (± 2), respectively. Median length of stay was 2 (range = 1– 3) days.Conclusion: Hypobaric opioid-free STSA appears to be a promising approach for laparoscopic surgeries, with minimal to null occurrence of shoulder pain. Larger prospective studies are required to validate these findings.Keywords: thoracic spinal anesthesia, laparoscopic cholecystectomy, hypobaric ropivacaine, intrathecal midazolam, intrathecal ketamine

Laboratory automation effectively increases the throughput in sample analysis, reduces human errors in sample processing, as well as simplifies and accelerates the overall logistics. Automating diagnostic testing workflows in peripheral laboratories and also in near-patient settings -like hospitals, clinics and epidemic control checkpoints- is advantageous for the simultaneous processing of multiple samples to provide rapid results to patients, minimize the possibility of contamination or error during sample handling or transport, and increase efficiency. However, most automation platforms are expensive and are not easily adaptable to new protocols. Here, we address the need for a versatile, easy-to-use, rapid and reliable diagnostic testing workflow by combining open-source modular automation (Opentrons) and automation-compatible molecular biology protocols, easily adaptable to a workflow for infectious diseases diagnosis by detection on paper-based diagnostics. We demonstrated the feasibility of automation of the method with a low-cost Neisseria meningitidis diagnostic test that utilizes magnetic beads for pathogen DNA isolation, isothermal amplification, and detection on a paper-based microarray. In summary, we integrated open-source modular automation with adaptable molecular biology protocols, which was also faster and cheaper to perform in an automated than in a manual way. This enables a versatile diagnostic workflow for infectious diseases and we demonstrated this through a low-cost N. meningitidis test on paper-based microarrays.

There is a growing interest for satellites complementing the terrestrial networks for broadband access service provision. One of the key challenges is represented by the need to match the non uniform on-ground traffic distribution. This calls for exploiting large frequency reuse among the satellite beams and the adoption of interference mitigation through the use of precoding and/or efficient radio resource management techniques. In recent years, space technological developments have been making possible the implementation of active antennas operating at 20–30 GHz with a good level of efficiency thus opening up new perspectives in the flexible system and payload design. The presence of a large number of antenna radiating elements on-board the satellite stimulates the application of Massive Multiple Input Multiple Output (M-MIMO) techniques to satellite networks. In this paper, we extend previous work related to (pragmatic) M-MIMO adaptations for satellite broadband access networks and associated affordable complexity Radio Resource Management (RRM) algorithms. In particular, we cover the case of frequency colouring scheme and the optimization of the precoding in presence of non uniform traffic. We also enhance the system model to achieve more realistic and comprehensive analysis of the performance results by means of the generation of a generic spatial traffic distribution, a more accurate satellite antenna modelling for the RRM algorithm and emulating the channel estimation errors. A spatial derivation of the traffic request system satisfaction factor has been developed to enhance the analysis of the simulation results. All these enhancements are applied to the previously published non uniform synthetically generated traffic cases and to a more realistic study case corresponding to a Geostationary Equatorial Orbit (GEO) satellite covering Americas for which a novel system optimization methodology has been developed.

Hardmetals are materials employed to produce cutting and forming tools as well as wear resistant parts. Standard powder metallurgy suffers limitations in the manufacturing of shapes required by application-oriented design. Therefore, there is interest for the implementation of additive manufacturing, in particular low temperature techniques as binder jetting because they allow to preserve microstructures and peculiar properties. In our work, the powder was tungsten carbide with 12 wt.% cobalt (WC-Co). Shaping by binder jetting was followed by curing treatment to promote the binder polymerization, vacuum sintering and sinter-HIP to achieve near-full density. The powder was analysed in terms of size and shape, to determine its suitability for the procedure. Then, the effects of different combinations of printing parameters, layer thickness (50–100 μm) and binder saturation (60/75/90%), on the density of the green bodies were analysed. Finally, the relative density of the sintered components was measured and the pores shape and size were studied by SEM imaging, to assess possible consequences of the deposition procedure. Vickers hardness along the samples cross-section was measured and correlated to the printing conditions employed.

Objective Coronavirus disease 2019 demonstrated the inconsistencies in adequately responding to biological threats on a global scale due to a lack of powerful tools for assessing various factors in the formation of the epidemic situation and its forecasting. Decision support systems have a role in overcoming the challenges in health monitoring systems in light of current or future epidemic outbreaks. This paper focuses on some applied examples of logistic planning, a key service of the Earth Cognitive System for Coronavirus Disease 2019 project, here presented, evidencing the added value of artificial intelligence algorithms towards predictive hypotheses in tackling health emergencies. Methods Earth Cognitive System for Coronavirus Disease 2019 is a decision support system designed to support healthcare institutions in monitoring, management and forecasting activities through artificial intelligence, social media analytics, geospatial analysis and satellite imaging. The monitoring, management and prediction of medical equipment logistic needs rely on machine learning to predict the regional risk classification colour codes, the emergency rooms attendances, and the forecast of regional medical supplies, synergically enhancing geospatial and temporal dimensions. Results The overall performance of the regional risk colour code classifier yielded a high value of the macro-average F1-score (0.82) and an accuracy of 85%. The prediction of the emergency rooms attendances for the Lazio region yielded a very low root mean square error (

Peculiar chemical, mechanical, and magnetic properties make cobalt a key metal for a variety of “hot” applications like the cathode production of Li-ion batteries. Cobalt is also the preferred metallic binder for tungsten carbide tool manufacturing. The recent increasing criticality of cobalt and tungsten is driving the interest of manufacturers and researchers toward high-rate recycling of hard-metal (HM) waste for limiting the demand for raw materials. A simple and environmentally friendly hydrometallurgical route for Co-selective dissolution from HM wastes was developed by using weak, bio-derived, and biodegradable organic acids (OAs). In this study, OAs, namely, acetic (HAc), citric (H3Cit), maleic (H2Mal), lactic (HLac), succinic (H2Suc), lactobionic (HLB), and itaconic (H2It) acids, were selected for their pKa1 values spanning from 1.8 to 4.7 and systematically tested as selective cobalt leaching agents from WC-Co-based wastes in water, isolating the formed complexes in the solid state. Thereby, all of them seemed to be efficient in selective Co leaching, achieving almost quantitative Co dissolution from HM by-products still at low concentration levels and room conditions in a short time, leaving the residual WC unreacted and ready to be re-employed for industrial purposes. Nevertheless, two main categories of organic acids were distinguished depending on their oxidizing/complexing behavior: class 1 OAs, where the metal oxidation is carried out by H+, and class 2 OAs, where oxidation is carried out by an external oxidant like O2. A combined experimental/theoretical investigation is described here to show the reasons behind this peculiar behavior and lay the foundation for a wider discussion on the leaching capabilities of OAs toward elemental metals. Due to the demonstrated effectiveness, low cost, eco-friendliness, and large availability through biotechnological fermentative processes, particular attention is devoted here to the use of HLac in hydrometallurgy as an example of class 2 OA. WC-Co materials recovered by HLac mild hydrometallurgy demonstrated a metallurgical quality suitable for re-employment in the HM manufacturing process.

This study investigates the influence of different sinter-HIP temperatures and binder saturation levels on the microstructure and properties of WC–12Co cemented carbide, produced using binder jetting. The sinter-HIP process was performed at 1400 °C, 1460 °C, and 1500 °C and binder saturation levels of 60% and 75% were selected during printing. The binder saturation proved to affect the repeatability of the manufacturing process and the sturdiness of the green models. The increase of the sintering temperature from 1400 °C to 1460 °C is correlated with an increase in the density. Nonetheless, a further raise in temperature to 1500 °C leads to significant grain coarsening without clear advantages in terms of porosity reduction. Both the transverse rupture strength and Vickers hardness increase when the sinter-HIP temperature rises from 1400 °C to 1460 °C, where the typical results for traditionally manufactured WC–12Co are met, with a comparable grain size. The transverse rupture strength and Vickers hardness then decrease for samples treated at 1500 °C. Finally, potential issues in the manufacturing process are identified and correlated with the defects in the final components.

BackgroundImpaired iron transport (IIT) is a form of iron deficiency (ID) defined as transferrin saturation (TSAT)

Paolo Vincenzi,1 Massimo Stronati,2 Paolo Isidori,1 Salvatore Iuorio,2 Diletta Gaudenzi,3 Gianfranco Boccoli,1 Roberto Starnari2 1Department of General Surgery, IRCSS-INRCA, Ancona, Italy; 2Department of Anaesthesiology, IRCSS-INRCA, Ancona, Italy; 3Department of Perioperative Services, AOU “Ospedali Riuniti di Ancona”, Ancona, ItalyCorrespondence: Paolo Vincenzi, Department of General Surgery, IRCSS-INRCA, Via della Montagnola n 81, Ancona, 60127, Italy, Tel + 39 3394578495, Email paolo.vincenzi1981@gmail.comPurpose: Few studies have described segmental thoracic spinal anesthesia (STSA) as primary anesthesiologic method in breast and axillary surgery, documenting the association of intrathecal local anesthetics and opioids. This case series reports an opioid-free scheme of STSA in four elderly patients undergoing major breast and axillary oncological surgery.Patients and Methods: STSA was performed in three female patients undergoing unilateral mastectomy ± axillary lymph node dissection (ALND) or sentinel lymph node biopsy for invasive ductal carcinoma and in one male patient undergoing ALND for melanoma metastases. The level of needle insertion was included between T6-8, via a median or paramedian approach. Midazolam (2 mg) and ketamine (20 mg) were used as adjuvants for intrathecal sedation, followed by the administration of hypobaric ropivacaine 0.25% at a dose of 8 mg. The level of sensory blockade achieved was comprised between C2-3 and T11-12. Postoperative analgesia was maintained through continuous intravenous administration of Ketorolac by an elastomeric pump (90 mg over 24 hrs.).Results: Spinal anesthesia was completed without complications in all patients. Conversion to general anesthesia (GA) and perioperative intravenous sedation were not required. No major postoperative complications and no episodes of postoperative nausea and vomiting (PONV) were reported. No rescue analgesic was administered. All patients were discharged in postoperative day 2 and are alive at 30, 29, 27 and 13 months after surgery, respectively. High grade of satisfaction on the anesthesiologic method was expressed by all cases.Conclusion: STSA with local anesthetic plus midazolam and ketamine might be considered a safe and effective alternative to GA, even in surgeries involving the breast and axillary region, particularly in elderly and frail patients. Larger prospective studies are required to validate these findings.Keywords: spinal anesthesia, breast cancer surgery, axillary surgery, intrathecal midazolam, intrathecal ketamine

Abstract Background Tanimilast is a novel and selective inhaled inhibitor of phosphodiesterase-4 in advanced clinical development for chronic obstructive pulmonary disease (COPD). Tanimilast is known to exert prominent anti-inflammatory activity when tested in preclinical experimental models as well as in human clinical studies. Recently, we have demonstrated that it also finely tunes, rather than suppressing, the cytokine network secreted by activated dendritic cells (DCs). This study was designed to characterize the effects of tanimilast on T-cell polarizing properties of DCs and to investigate additional functional and phenotypical features induced by tanimilast. Methods DCs at day 6 of culture were stimulated with LPS in the presence or absence of tanimilast or the control drug budesonide. After 24 h, DCs were analyzed for the expression of surface markers of maturation and activation by flow cytometry and cocultured with T cells to investigate cell proliferation and activation/polarization. The regulation of type 2-skewing mediators was investigated by real-time PCR in DCs and compared to results obtained in vivo in a randomized placebo-controlled trial on COPD patients treated with tanimilast. Results Our results show that both tanimilast and budesonide reduced the production of the immunostimulatory cytokine IFN-γ by CD4+ T cells. However, the two drugs acted at different levels since budesonide mainly blocked T cell proliferation, while tanimilast skewed T cells towards a Th2 phenotype without affecting cell proliferation. In addition, only DCs matured in the presence of tanimilast displayed increased CD86/CD80 ratio and CD141 expression, which correlated with Th2 T cell induction and dead cell uptake respectively. These cells also upregulated cAMP-dependent immunosuppressive molecules such as IDO1, TSP1, VEGF-A and Amphiregulin. Notably, the translational value of these data was confirmed by the finding that these same genes were upregulated also in sputum cells of COPD patients treated with tanimilast as add-on to inhaled glucocorticoids and bronchodilators. Conclusion Taken together, these findings demonstrate distinct immunomodulatory properties of tanimilast associated with a type 2 endotype and CD141 upregulation in DCs and provide a mechanistic rationale for the administration of tanimilast on top of inhaled corticosteroids.

Abstract Invasive nonfunctioning (NF) pituitary neuroendocrine tumors (PitNETs) are non‐resectable neoplasms associated with frequent relapses and significant comorbidities. As the current therapies of NF‐PitNETs often fail, new therapeutic targets are needed. The observation that circulating angiopoietin‐2 (ANGPT2) is elevated in patients with NF‐PitNET and correlates with tumor aggressiveness prompted us to investigate the ANGPT2/TIE2 axis in NF‐PitNETs in the GH3 PitNET cell line, primary human NF‐PitNET cells, xenografts in zebrafish and mice, and in MENX rats, the only autochthonous NF‐PitNET model. We show that PitNET cells express a functional TIE2 receptor and secrete bioactive ANGPT2, which promotes, besides angiogenesis, tumor cell growth in an autocrine and paracrine fashion. ANGPT2 stimulation of TIE2 in tumor cells activates downstream cell proliferation signals, as previously demonstrated in endothelial cells (ECs). Tie2 gene deletion blunts PitNETs growth in xenograft models, and pharmacological inhibition of Angpt2/Tie2 signaling antagonizes PitNETs in primary cell cultures, tumor xenografts in mice, and in MENX rats. Thus, the ANGPT2/TIE2 axis provides an exploitable therapeutic target in NF‐PitNETs and possibly in other tumors expressing ANGPT2/TIE2. The ability of tumor cells to coopt angiogenic signals classically viewed as EC‐specific expands our view on the microenvironmental cues that are essential for tumor progression.