Interaction between NSCLC Cells, CD8+ T-Cells and Immune Checkpoint Inhibitors Potentiates Coagulation and Promotes Metabolic Remodeling-New Cues on CAT-VTE.

BACKGROUND: Cancer-associated thrombosis (CAT) and venous thromboembolism (VTE) are frequent cancer-related complications associated with high mortality; thus, this urges the identification of predictive markers. Immune checkpoint inhibitors (ICIs) used in cancer immunotherapy allow T-cell activation against cancer cells. Retrospective studies showed increased VTE following ICI administration in some patients. Non-small cell lung cancer (NSCLC) patients are at high risk of thrombosis and thus, the adoption of immunotherapy, as a first-line treatment, seems to be associated with coagulation-fibrinolysis derangement. METHODS: We pharmacologically modulated NSCLC cell lines in co-culture with CD8+ T-cells (TCD8+) and myeloid-derived suppressor cells (MDSCs), isolated from healthy blood donors. The effects of ICIs Nivolumab and Ipilimumab on NSCLC cell death were assessed by annexin V and propidium iodide (PI) flow cytometry analysis. The potential procoagulant properties were analyzed by in vitro clotting assays and enzyme-linked immunosorbent assays (ELISAs). The metabolic remodeling induced by the ICIs was explored by 1H nuclear magnetic resonance (NMR) spectroscopy. RESULTS: Flow cytometry analysis showed that TCD8+ and ICIs increase cell death in H292 and PC-9 cells but not in A549 cells. Conditioned media from NSCLC cells exposed to TCD8+ and ICI induced in vitro platelet aggregation. In A549, Podoplanin (PDPN) levels increased with Nivolumab. In H292, ICIs increased PDPN levels in the absence of TCD8+. In PC-9, Ipilimumab decreased PDPN levels, this effect being rescued by TCD8+. MDSCs did not interfere with the effect of TCD8+ in the production of TF or PDPN in any NSCLC cell lines. The exometabolome showed a metabolic remodeling in NSCLC cells upon exposure to TCD8+ and ICIs. CONCLUSIONS: This study provides some insights into the interplay of immune cells, ICIs and cancer cells influencing the coagulation status. ICIs are important promoters of coagulation, benefiting from TCD8+ mediation. The exometabolome analysis highlighted the relevance of acetate, pyruvate, glycine, glutamine, valine, leucine and isoleucine as biomarkers. Further investigation is needed to validate this finding in a cohort of NSCLC patients.

Electrochemical immunosensor for point-of-care detection of soybean Gly m TI allergen in foods.

Soybean is a legume with high technological functionality, commonly used by the food industry as an ingredient in different products. However, soybean is an allergenic food whose undeclared presence in processed foods may represent a public health risk. In this work, it was developed an efficient electrochemical immunosensor, targeting the soybean trypsin inhibitor (Gly m TI) allergen using commercial anti-Gly m TI IgG, aiming at detecting/quantifying minute amounts of soybean in different food formulations. For this purpose, model mixtures of different foods (sausages, cooked-hams, biscuits) were prepared to contain known amounts of soybean protein isolate (100,000-0.1 mg kg-1) and submitted to specific thermal treatments (autoclaving, oven-cooking, baking). The electrochemical immunosensor allowed quantifying down to 0.1 mg kg-1 of soybean in the three food matrices, raw and processed (0.0012 mg of Gly m TI/kg of matrix). Accordingly, the immunosensor is suitable for detecting traces of soybean in raw, processed, and complex foods, thus protecting 99 % of soybean-allergic patients.

MOF-808 as an Efficient Catalyst for Valorization of Biodiesel Waste Production: Glycerol Acetalization.

Glycerol is the main residue in the biodiesel production industry; therefore, their valorization is crucial. The acetalization of glycerol toward fuel additives such as solketal (2,2-dimethyl-1,3-dioxolan-4-methanol) is of high interest, promoting circular economy since it can be added to biodiesel or even fossil diesel to improve their quality and efficiency. Straightforward-prepared metal-organic framework (MOF) materials of the MOF-808 family were applied to the valorization of glycerol for the first time. In particular, MOF-808(Hf) was revealed to be an effective heterogeneous catalyst to produce solketal under moderate conditions: a small amount of the MOF material (only 4 wt% of glycerol), a 1:6 ratio of glycerol/acetone, and a temperature of 333 K. The high efficiency of MOF-808(Hf) was associated with the high amount of acid centers present in its structure. Furthermore, its structural characteristics, such as window opening cavity size and pore diameters, were shown to be ideal for reusing this material for at least ten consecutive reaction cycles without losing activity (conversion > 90% and selectivity > 98%). Remarkably, it was not necessary to wash or activate the MOF-808(Hf) catalyst between cycles (no pore blockage occurred), and it maintained structural integrity after ten cycles, confirming its ability to be a sustainable heterogeneous catalyst for glycerol valorization.

BIBBOX, a FAIR toolbox and App Store for life science research.

Data quality has recently become a critical topic for the research community. European guidelines recommend that scientific data should be made FAIR: findable, accessible, interoperable and reusable. However, as FAIR guidelines do not specify how the stated principles should be implemented, it might not be straightforward for researchers to know how actually to make their data FAIR. This can prevent life-science researchers from sharing their datasets and pipelines, ultimately hindering the progress of research. To address this difficulty, we developed the BIBBOX, which is a platform that supports researchers publishing their datasets and the associated software in a FAIR manner.

Engineering a membrane-binding protein to trimerize and induce high membrane curvature.

The annexins are a family of Ca2+-dependent peripheral membrane proteins. Several annexins are implicated in plasma membrane repair and are overexpressed in cancer cells. Annexin A4 (ANXA4) and annexin A5 (ANXA5) form trimers that induce high curvature on a membrane surface, a phenomenon deemed to accelerate membrane repair. Despite being highly homologous to ANXA4, annexin A3 (ANXA3) does not form trimers on the membrane surface. Using molecular dynamics simulations, we have reverse engineered an ANXA3-mutant to trimerize on the surface of the membrane and induce high curvature reminiscent of ANXA4. In addition, atomic force microscopy images show that, like ANXA4, the engineered protein forms crystalline arrays on a supported lipid membrane. Despite the trimer-forming and curvature-inducing properties of the engineered ANXA3, it does not accumulate near a membrane lesion in laser-punctured cells and is unable to repair the lesion. Our investigation provides insights into the factors that drive annexin-mediated membrane repair and shows that the membrane-repairing property of trimer-forming annexins also necessitates high membrane binding affinity, other than trimer formation and induction of negative membrane curvature.

Plasmonic genosensor for detecting hazelnut Cor a 14-encoding gene for food allergen monitoring.

A plasmonic nanostructure was constructed as a biorecognition element coupled to an optical sensing platform in sandwich format, targeting the hazelnut Cor a 14 allergen-encoding gene. The analytical performance of the genosensor presented a linear dynamic range between 100 amol L-1 and 1 nmol L-1, a limit of detection (LOD) < 19.9 amol L-1, and a sensitivity of 13.4 ± 0.6 m°. The genosensor was successfully hybridized with hazelnut PCR products, tested with model foods, and further validated by real-time PCR. It reached a LOD <0.001% (10 mg kg-1) of hazelnut in wheat material (corresponding to 1.6 mg kg-1 of protein) and a sensitivity of -17.2 ± 0.5 m° for a linear range of 0.001%-1%. Herein, a new genosensing approach is proposed as a highly sensitive and specific alternative tool with potential application in monitoring hazelnut as an allergenic food, protecting the health of sensitized/allergic individuals.

Chitins from Seafood Waste as Sustainable Porous Carbon Precursors for the Development of Eco-Friendly Supercapacitors.

Carbon materials derived from marine waste have been drawing attention for supercapacitor applications. In this work, chitins from squid and prawn marine wastes were used as carbon precursors for further application as electrodes for energy storage devices. Chitins were obtained through a deproteinization method based on enzymatic hydrolysis as an alternative to chemical hydrolysis as commonly presented in the literature. The obtained porous carbons were characterized using a BET surface area analyzer to determine the specific surface area and pore size, as well as scanning electron microscopy (SEM) with energy dispersive X-ray analysis (EDX), transmission electron microscopy (TEM), Raman spectroscopy, attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS), to characterize their morphology, composition, and structure. The electrochemical characterization was performed using a glassy carbon (GC) electrode modified with marine waste-based porous carbons as the working electrode through cyclic voltammetry and galvanostatic charge/discharge using ethaline, a choline chloride-based deep eutectic solvent (DES), as an eco-friendly and sustainable electrolyte. Squid and prawn chitin-based carbons presented a surface area of 149.3 m2 g-1 and 85.0 m2 g-1, pore volume of 0.053 cm3 g-1 and 0.029 cm3 g-1, and an associated specific capacitance of 20 and 15 F g-1 at 1 A g-1, respectively. Preliminary studies were performed to understand the effect of -OH groups on the chitin-based carbon surface with DES as an electrolyte, as well as the effect of aqueous electrolytes (1 mol L-1 sulphuric acid (H2SO4) and 1 mol L-1 potassium hydroxide (KOH)) on the capacitance and retention of the half-cell set up. It is provided, for the first time, the use of chitin-based carbon materials obtained through a one-step carbonization process combined with an eco-friendly DES electrolyte for potential application in energy storage devices.

Promoção da literacia e saúde mental positiva: Estudo quase-experimental com adolescentes em contexto esco / Promotion of literacy and positive mental health: A quasi-experimental study with adolescents in a school context

Introdução: As intervenções de enfermagem de promoção da saúde mental positiva (SMP)em adolescentes aumentam a autoestima, as habilidades socio emocionais e a capacidade de resolução de problemas dos jovens, diminuem o estigma, e permitem incrementar a esperança. Objetivos: Analisar uma intervenção de promoção de saúde mental positiva em adolescentes em contexto escolar. Metodologia: Estudo quase-experimental com pré e pós-teste, numa amostra não probabilística de 72 adolescentes. Foi aplicado um programa de SMP por enfermeiros especialistas de saúde mental no grupo experimental. Para a recolha de dados foi usado questionário de autopreenchimento online que continha todos os instrumentos de medida, aplicados ao grupo experimental antes e depois da intervenção de promoção de saúde mental positiva. Resultados: Verificou-se melhoria da SMP, diminuição do estigma, diminuição da vulnerabilidade psicológica e aumento dos níveis de esperança. Conclusão: A intervenção de enfermagem de promoção da saúde mental positiva teve uma forte adesão dos adolescentes e uma melhoria nos indicadores de saúde mental estudados (AU)

Introduction: Nursing interventions to promote positive mental health (PMH) in adolescents increase self-esteem socio-emotional and problem-solving skills of young people, reduce stigma and increase hope. Objective: To analyse the programme’s effectiveness in promoting adolescents’ positive mental health in a school context. Methodology: A Quasi-experimental study using pre and post-test in a non-probabilistic sample of 72 adolescents. A PMH programme was applied by specialist mental health nurses in the experimental group. An online self-completion questionnaire was used for data collection, which contained the measurement instruments applied to the experimental group before and after the PMH intervention. Results: There was an improvement in PMH, a decrease in stigma, a decrease in psychological vulnerability and an increase in levels of hope. Conclusion: The positive mental health promotion nursing intervention had a strong adherence by adolescents and improved the mental health indicators under study. (AU)

Annexin A7 mediates lysosome repair independently of ESCRT-III.

Lysosomes are crucial organelles essential for various cellular processes, and any damage to them can severely compromise cell viability. This study uncovers a previously unrecognized function of the calcium- and phospholipid-binding protein Annexin A7 in lysosome repair, which operates independently of the Endosomal Sorting Complex Required for Transport (ESCRT) machinery. Our research reveals that Annexin A7 plays a role in repairing damaged lysosomes, different from its role in repairing the plasma membrane, where it facilitates repair through the recruitment of ESCRT-III components. Notably, our findings strongly suggest that Annexin A7, like the ESCRT machinery, is dispensable for membrane contact site formation within the newly discovered phosphoinositide-initiated membrane tethering and lipid transport (PITT) pathway. Instead, we speculate that Annexin A7 is recruited to damaged lysosomes and promotes repair through its membrane curvature and cross-linking capabilities. Our findings provide new insights into the diverse mechanisms underlying lysosomal membrane repair and highlight the multifunctional role of Annexin A7 in membrane repair.

Memristor-Based Neuromodulation Device for Real-Time Monitoring and Adaptive Control of Neuronal Populations.

Neurons are specialized cells for information transmission and information processing. In fact, many neurologic disorders are directly linked not to cellular viability/homeostasis issues but rather to specific anomalies in electrical activity dynamics. Consequently, therapeutic strategies based on the direct modulation of neuronal electrical activity have been producing remarkable results, with successful examples ranging from cochlear implants to deep brain stimulation. Developments in these implantable devices are hindered, however, by important challenges such as power requirements, size factor, signal transduction, and adaptability/computational capabilities. Memristors, neuromorphic nanoscale electronic components able to emulate natural synapses, provide unique properties to address these constraints, and their use in neuroprosthetic devices is being actively explored. Here, we demonstrate, for the first time, the use of memristive devices in a clinically relevant setting where communication between two neuronal populations is conditioned to specific activity patterns in the source population. In our approach, the memristor device performs a pattern detection computation and acts as an artificial synapse capable of reversible short-term plasticity. Using in vitro hippocampal neuronal cultures, we show real-time adaptive control with a high degree of reproducibility using our monitor-compute-actuate paradigm. We envision very similar systems being used for the automatic detection and suppression of seizures in epileptic patients.

Clinical questions in primary care: Where to find the answers - a cross-sectional study.

INTRODUCTION: Clinicians raise at least one question for every two patients they see, but search for an answer to less than half of these questions, and rarely use evidence-based resources. One barrier to evidence-based practice is doubt that the search would yield an answer, and we found insufficient evidence to refute this concern. This study aims to identify what proportion of clinical questions in primary care can be answered with online evidence-based practice resources, and what proportion of these can be answered with pre-appraised evidence. MATERIALS AND METHODS: Cross-sectional study in two primary care practices. The inclusion criteria were family doctors, generalists and residents working in 2 selected practices. We collected a total of 238 questions from 19 family medicine specialists, 9 family medicine residents and 3 generalist doctors. Doctors were asked to record any clinical question that arose during 4 days of appointments. The primary outcome was the proportion of clinical questions answered with online evidence-based practice resources. The secondary outcome was the level of evidence needed to reach to find the answers (clinical summaries, systematic guidelines, systematic reviews or primary studies), according to Haynes' pre-appraised evidence pyramid model. RESULTS: 191 of the 206 valid clinical questions could be answered with online evidence-based practice resources (92.7% [95% CI 88.3%-95.9%]). Most of these questions (90.8% CI 95% 85.9%-94.4%) were successfully answered using clinical summaries (BMJ Best Practice, DynaMed or UpToDate), with a median search time of 4 minutes (range 1-16.5). CONCLUSIONS: Contrary to clinician's beliefs, the majority of clinical questions can be answered with online evidence-based practice resources, and most of them with pre-appraised evidence. This study could encourage family doctors to increase the use of clinical summaries. Furthermore, these results highlight the importance of teaching how to search for and apply pre-appraised evidence.

Plasma Membrane Wounding and Repair Assays for Eukaryotic Cells.

Damage to the plasma membrane and loss of membrane integrity are detrimental to eukaryotic cells. It is, therefore, essential that cells possess an efficient membrane repair system to survive. However, the different cellular and molecular mechanisms behind plasma membrane repair have not been fully elucidated. Here, we present three complementary methods for plasma membrane wounding, and measurement of membrane repair and integrity. The first protocol is based on real time imaging of cell membrane repair kinetics in response to laser-induced injury. The second and third protocols are end point assays that provide a population-based measure of membrane integrity, after either mechanical injury by vortex mixing with glass beads, or by detergent-induced injury by digitonin in sublytic concentrations. The protocols can be applied to most adherent eukaryotic cells in culture, as well as cells in suspension.

Feel Your Reach: An EEG-Based Framework to Continuously Detect Goal-Directed Movements and Error Processing to Gate Kinesthetic Feedback Informed Artificial Arm Control.

Establishing the basic knowledge, methodology, and technology for a framework for the continuous decoding of hand/arm movement intention was the aim of the ERC-funded project "Feel Your Reach". In this work, we review the studies and methods we performed and implemented in the last 6 years, which build the basis for enabling severely paralyzed people to non-invasively control a robotic arm in real-time from electroencephalogram (EEG). In detail, we investigated goal-directed movement detection, decoding of executed and attempted movement trajectories, grasping correlates, error processing, and kinesthetic feedback. Although we have tested some of our approaches already with the target populations, we still need to transfer the "Feel Your Reach" framework to people with cervical spinal cord injury and evaluate the decoders' performance while participants attempt to perform upper-limb movements. While on the one hand, we made major progress towards this ambitious goal, we also critically discuss current limitations.

Generation of a CHIP isogenic human iPSC-derived cortical neuron model for functional proteomics.

The neuroprotective E3-ubiquitin ligase CHIP is linked to healthy aging. Here, we present a protocol using a patient-derived iPSC line with a triplication of the α-synuclein gene to produce gene-edited cells isogenic for CHIP. We describe iPSC differentiation into cortical neurons and their identity validation. We then detail mass spectrometry-based approaches (SWATH-MS) to identify dominant changes in the steady state proteome generated by loss of CHIP function. This protocol can be adapted to other proteins that impact proteostasis in neurons. For complete details on the use and execution of this protocol, please refer to Dias et al. (2021).

Erratum: CHIP-dependent regulation of the actin cytoskeleton is linked to neuronal cell membrane integrity.

[This corrects the article DOI: 10.1016/j.isci.2021.102878.].

A novel NINJ1-mediated regulatory step is essential for active membrane rupture and common to different cell death pathways.

Plasma membrane rupture (PMR), the final event in lytic cell death that is in part responsible for the release of pro-inflammatory signals, was believed to be a passive event that followed osmotic swelling. Kayagaki et al. 1 have discovered that PMR is, in fact, mediated by ninjurin-1 (NINJ1), adding a novel regulatory step that is conserved across different types of lytic cell death, such as pyroptosis, necroptosis, and apoptosis. PMR is dependent on NINJ1 oligomerization, which is mediated by its highly conserved putative N-terminal α-helix. In vivo data suggest that the NINJ1-dependent secretome that is released upon PMR is likely to modulate antimicrobial host defense, suggesting this additional regulatory step also has physiological relevance.

Prognostic Value of the Malnutrition-inflammation Score in Hospitalization and Mortality on Long-term Hemodialysis.

OBJECTIVE: Since its development, cumulative evidence has accumulated regarding the prognostic value of the Malnutrition-Inflammation Score (MIS/Kalantar score) prognostic value; however, there is a shortage of recent and large studies with comprehensive statistical methodologies that contribute to support a higher level of evidence and a consensual cutoff. The aim of this study was to assess the strength of MIS association with hospitalization and mortality in a nationwide cohort. METHODS: This was a historical cohort study of hemodialysis patients from 25 outpatient centers followed up for 48 months. Univariable and multivariable Cox additive regression models were used to analyze the data. The C-index was estimated to assess the performance of the final model. RESULTS: Two thousand four hundred forty-four patients were analyzed, 59.0% males, 32.0% diabetic, and median age of 71 years (P25 = 60, P75 = 79). During a median period of 45-month follow-up, with a maximum of 48 months (P25 = 31; P75 = 48), 875 patients presented an MIS <5 (35.8%) and 860 patients (35.2%) died. The proportion of deaths was 23.1% for patients with the MIS <5 and 41.9% if the MIS ≥5 (P < .001). A total of 1,528 patients (62.5%) were hospitalized with a median time to the first hospitalization of 26 months (P25 = 9; P75 = 45). A new cutoff point regarding the risk of death, MIS ≥6, was identified for this study data set. In multivariable analysis for hospitalization risk, a higher MIS, higher comorbidity index, and arteriovenous graft or catheter increased the risk, whereas higher Kt/V and higher albumin had a protective effect. In multivariable analysis for mortality risk, adjusting for age, albumin, normalized protein catabolic rate, Charlson comorbidity index, interdialytic weight gain, Kt/V, diabetes, hematocrit, and vascular access, patients with the MIS ≥6 showed a hazard ratio of 1.469 (95% confidence interval: 1.262-1.711; P < .001). Higher age, higher interdialytic weight gain, higher comorbidity index, and catheter increased significantly the risk, whereas higher Kt/V, higher albumin, and higher normalized protein catabolic rate (≥1.05 g/kg/d) reduced the risk. CONCLUSION: The MIS maintains its relevant and significant association with hospitalization and mortality.