Multiphysics Finite\u2013Element Modelling of an All\u2013Vanadium Redox Flow Battery for Stationary Energy Storage

All-Vanadium Redox Flow Batteries (VRFBs) are emerging as a novel technology for stationary energy storage. Numerical models are useful for exploring the potential performance of such devices, optimizing the structure and operating condition of cell stacks, and studying its interfacing to the electrical grid. A one-dimensional steady-state multiphysics model of a single VRFB, including mass, charge and momentum transport and conservation, and coupled to a kinetic model for electrochemical reactions, is first presented. This model is then extended, including reservoir equations, in order to simulate the VRFB charge and discharge dynamics. These multiphysics models are discretized by the finite element method in a commercial software package (COMSOL). Numerical results of both static and dynamic 1D models are compared to those from 2D models, with the same parameters, showing good agreement. This motivates the use of reduced models for a more efficient system simulation

Patient-Derived Xenografts of Non Small Cell Lung Cancer: Resurgence of an Old Model for Investigation of Modern Concepts of Tailored Therapy and Cancer Stem Cells

Current chemotherapy regimens have unsatisfactory results in most advanced solid tumors. It is therefore imperative to devise novel therapeutic strategies and to optimize selection of patients, identifying early those who could benefit from available treatments. Mouse models are the most valuable tool for preclinical evaluation of novel therapeutic strategies in cancer and, among them, patient-derived xenografts models (PDX) have made a recent comeback in popularity. These models, obtained by direct implants of tissue fragments in immunocompromised mice, have great potential in drug development studies because they faithfully reproduce the patient's original tumor for both immunohistochemical markers and genetic alterations as well as in terms of response to common therapeutics They also maintain the original tumor heterogeneity, allowing studies of specific cellular subpopulations, including their modulation after drug treatment. Moreover PDXs maintain at least some aspects of the human microenvironment for weeks with the complete substitution with murine stroma occurring only after 2-3 passages in mouse and represent therefore a promising model for studies of tumor-microenvironment interaction. This review summarizes our present knowledge on mouse preclinical cancer models, with a particular attention on patient-derived xenografts of non small cell lung cancer and their relevance for preclinical and biological studies

Generation of functional HLA-DR*1101 tetramers receptive for loading with pathogen or tumour derived synthetic peptides

BACKGROUND: MHC class I-peptide tetramers are currently utilised to characterize CD8(+ )T cell responses at single cell level. The generation and use of MHC class II tetramers to study antigen-specific CD4(+ )T cells appears less straightforward. Most MHC class II tetramers are produced with a homogeneously built-in peptide, reducing greatly their flexibility of use. We attempted the generation of "empty" functional HLA-DR*1101 tetramers, receptive for loading with synthetic peptides by incubation. No such reagent is in fact available for this HLA-DR allele, one of the most frequent in the Caucasian population. RESULTS: We compared soluble MHC class II-immunoglobulin fusion proteins (HLA-DR*1101-Ig) with soluble MHC class II protein fused with an optimised Bir site for enzymatic biotynilation (HLA-DR*1101-Bir), both produced in insect cells. The molecules were multimerised by binding fluorochrome-protein A or fluorochrome-streptavidin, respectively. We find that HLA-DR*1101-Bir molecules are superior to the HLA-DR*1101-Ig ones both in biochemical and functional terms. HLA-DR*1101-Bir molecules can be pulsed with at least three different promiscuous peptide epitopes, derived from Tetanus Toxoid, influenza HA and the tumour associated antigen MAGE-3 respectively, to stain specific CD4(+ )T cells. Both staining temperature and activation state of CD4(+ )T cells are critical for the binding of peptide-pulsed HLA-DR*1101-Bir to the cognate TCR. CONCLUSION: It is therefore possible to generate a soluble recombinant HLA-DR*1101 backbone that is receptive for loading with different peptides to stain specific CD4(+ )T cells. As shown for other HLA-DR alleles, we confirm that not all the strategies to produce soluble HLA-DR*1101 multimers are equivalent

Identification and Characterization of a Novel Family of Cysteine-Rich Peptides (MgCRP-I) from Mytilus galloprovincialis

We report the identification of a novel gene family (named MgCRP-I) encoding short secreted cysteine-rich peptides in the Mediterranean mussel Mytilus galloprovincialis. These peptides display a highly conserved pre-pro region and a hypervariable mature peptide comprising six invariant cysteine residues arranged in three intramolecular disulfide bridges. Although their cysteine pattern is similar to cysteines-rich neurotoxic peptides of distantly related protostomes such as cone snails and arachnids, the different organization of the disulfide bridges observed in synthetic peptides and phylogenetic analyses revealed MgCRP-I as a novel protein family. Genome- and transcriptome-wide searches for orthologous sequences in other bivalve species indicated the unique presence of this gene family in Mytilus spp. Like many antimicrobial peptides and neurotoxins, MgCRP-I peptides are produced as pre-propeptides, usually have a net positive charge and likely derive from similar evolutionary mechanisms, that is, gene duplication and positive selection within the mature peptide region; however, synthetic MgCRP-I peptides did not display significant toxicity in cultured mammalian cells, insecticidal, antimicrobial, or antifungal activities. The functional role of MgCRP-I peptides in mussel physiology still remains puzzling

PROPOSTA METODOLÓGICA PARA DEFINIÇÃO DE ÁREAS PRIORITÁRIAS PARA RECUPERAÇÃO VEGETAL DE ÁREAS DE PRESERVAÇÃO PERMANENTE

O crescimento populacional e a expansão urbana ocasionam fortes pressões nas áreas naturais, resultando na substituição das florestas e degradação ambiental. Assim, o estudo objetivou a proposição de uma metodologia para identificação e definição de áreas prioritárias para recuperação vegetal de Áreas de Preservação Permanente (APP) da Área de Proteção Ambiental (APA) do Ribeirão Engenho d’Água, localizada no município de Porto Feliz, SP. A metodologia consistiu na elaboração de diferentes planos de informação e análise multicritério apoiadas em ambiente de Sistema de Informação Geográfica e Sensoriamento Remoto. Os resultados revelaram que 74,52% das áreas de APP necessitam de recuperação ambiental. Quanto à declividade, 15,32% foi classificada como forte ou média nos extremos norte e sul, e na região central 84,68% variaram de fracas a muito fracas. Os solos apresentaram muito fraca erodibilidade em 72,9% da APA. O uso do solo revelou que 84,53% das áreas que necessitam de recuperação são ocupadas por culturas agrícolas; 7,11% por vegetação arbórea-arbustiva; 3,25% por áreas de várzea e pastagem; e pasto sujo totalizaram 3,94%. Em síntese, as áreas que apresentaram média e alta prioridade de recomposição das APPs compõem 9,34% da área de estudo, possuindo, em grande parte, solo do tipo argissolo vermelho e amarelo classificado com potencial de erosão médio a alto, e valores de declividade médios com elevação entre 12 a 20%. A proposta metodológica, auxiliada por geotecnologia, apresentou resultados bastante satisfatórios, podendo ser aplicada no diagnóstico ambiental de bacias hidrográficas com vistas à proteção dos recursos hídricos.

Assessment of spatio-temporal variability of faecal pollution along coastal waters during and after rainfall events

© The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Manini, E., Baldrighi, E., Ricci, F., Grilli, F., Giovannelli, D., Intoccia, M., Casabianca, S., Capellacci, S., Marinchel, N., Penna, P., Moro, F., Campanelli, A., Cordone, A., Correggia, M., Bastoni, D., Bolognini, L., Marini, M., & Penna, A. Assessment of spatio-temporal variability of faecal pollution along coastal waters during and after rainfall events. Water, 14(3), (2022): 502, https://doi.org/10.3390/w14030502.More than 80% of wastewaters are discharged into rivers or seas, with a negative impact on water quality along the coast due to the presence of potential pathogens of faecal origin. Escherichia coli and enterococci are important indicators to assess, monitor, and predict microbial water quality in natural ecosystems. During rainfall events, the amount of wastewater delivered to rivers and coastal systems is increased dramatically. This study implements measures capable of monitoring the pathways of wastewater discharge to rivers and the transport of faecal bacteria to the coastal area during and following extreme rainfall events. Spatio-temporal variability of faecal microorganisms and their relationship with environmental variables and sewage outflow in an area located in the western Adriatic coast (Fano, Italy) was monitored. The daily monitoring during the rainy events was carried out for two summer seasons, for a total of five sampling periods. These results highlight that faecal microbial contaminations were related to rainy events with a high flow of wastewater, with recovery times for the microbiological indicators varying between 24 and 72 h and influenced by a dynamic dispersion. The positive correlation between ammonium and faecal bacteria at the Arzilla River and the consequences in seawater can provide a theoretical basis for controlling ammonium levels in rivers as a proxy to monitor the potential risk of bathing waters pathogen pollution.This research was funded by WATERCARE project (Water management solutions for reducing microbial environment impact in coastal areas, project ID 10044130, https://www.italy-croatia.eu/web/watercare, accessed on 17 October 2021) funded by the European Union under the Interreg Italy–Croatia CBC Programme

LKB1 Down-Modulation by miR-17 Identifies Patients With NSCLC Having Worse Prognosis Eligible for Energy-Stress–Based Treatments

Abstract Introduction Preclinical models recently unveiled the vulnerability of LKB1/KRAS comutated NSCLC to metabolic stress-based treatments. Because miR-17 is a potential epigenetic regulator of LKB1, we hypothesized that wild-type LKB1 (LKB1WT) NSCLC with high miR-17 expression may be sensitive to an energetic stress condition, and eligible for metabolic frailties-based therapeutic intervention. Methods We took advantage of NSCLC cell lines with different combinations of KRAS mutation and LKB1 deletion and of patient-derived xenografts (PDXs) with high (LKB1WT/miR-17 high) or low (LKB1WT/miR-17 low) miR-17 expression. We evaluated LKB1 pathway impairment and apoptotic response to metformin. We retrospectively evaluated LKB1 and miR-17 expression levels in tissue specimens of patients with NSCLC and PDXs. In addition, a lung cancer series from The Cancer Genome Atlas data set was analyzed for miR-17 expression and potential correlation with clinical features. Results We identified miR-17 as an epigenetic regulator of LKB1 in NSCLC and confirmed targeting of miR-17 to LKB1 3′ untranslated region by luciferase reporter assay. We found that miR-17 overexpression functionally impairs the LKB1/AMPK pathway. Metformin treatment prompted apoptosis on miR-17 overexpression only in LKB1WT cell lines, and in LKB1WT/miR-17 high PDXs. A retrospective analysis in patients with NSCLC revealed an inverse correlation between miR-17 and LKB1 expression and highlighted a prognostic role of miR-17 expression in LKB1WT patients, which was further confirmed by The Cancer Genome Atlas data analysis. Conclusions We identified miR-17 as a mediator of LKB1 expression in NSCLC tumors. This study proposes a miR-17 expression score potentially exploitable to discriminate LKB1WT patients with NSCLC with impaired LKB1 expression and poor outcome, eligible for energy-stress-based treatments

Metformin Enhances Cisplatin-Induced Apoptosis and Prevents Resistance to Cisplatin in Co-mutated KRAS/LKB1 NSCLC

Abstract Introduction We hypothesized that activating KRAS mutations and inactivation of the liver kinase B1 (LKB1) oncosuppressor can cooperate to sustain NSCLC aggressiveness. We also hypothesized that the growth advantage of KRAS/LKB1 co-mutated tumors could be balanced by higher sensitivity to metabolic stress conditions, such as metformin treatment, thus revealing new strategies to target this aggressive NSCLC subtype. Methods We retrospectively determined the frequency and prognostic value of KRAS/LKB1 co-mutations in tissue specimens from NSCLC patients enrolled in the TAILOR trial. We generated stable LKB1 knockdown and LKB1-overexpressing isogenic H1299 and A549 cell variants, respectively, to test the in vitro efficacy of metformin. We also investigated the effect of metformin on cisplatin-resistant CD133+ cells in NSCLC patient-derived xenografts. Results We found a trend towards worse overall survival in patients with KRAS/LKB1 co-mutated tumors as compared to KRAS-mutated ones (hazard ratio: 2.02, 95% confidence interval: 0.94–4.35, p = 0.072). In preclinical experiments, metformin produced pro-apoptotic effects and enhanced cisplatin anticancer activity specifically in KRAS/LKB1 co-mutated patient-derived xenografts. Moreover, metformin prevented the development of acquired tumor resistance to 5 consecutive cycles of cisplatin treatment (75% response rate with metformin-cisplatin as compared to 0% response rate with cisplatin), while reducing CD133+ cells. Conclusions LKB1 mutations, especially when combined with KRAS mutations, may define a specific and more aggressive NSCLC subtype. Metformin synergizes with cisplatin against KRAS/LKB1 co-mutated tumors, and may prevent or delay the onset of resistance to cisplatin by targeting CD133+ cancer stem cells. This study lays the foundations for combining metformin with standard platinum-based chemotherapy in the treatment of KRAS/LKB1 co-mutated NSCLC

Metformin +/- cyclic fasting mimicking diet in combination with platinum-pemetrexed chemotherapy for advanced LKB1 inactive lung adenocarcinoma: The FAME trial

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