IntroductionHeart failure with preserved ejection fraction (HFpEF) accounts for nearly half of all heart failure cases and lacks effective therapies. Key features of HFpEF include endothelial dysfunction, fibrosis, and oxidative stress. Adenosine signaling, regulated by enzymes and receptors, is critical for vascular homeostasis and inflammation, but its role in HFpEF remains poorly understood. Adenosine receptors are abundantly expressed in the heart and kidney, modulating vascular, fibrotic, and tubular processes. Dysregulation of adenosine pathways in either organ may drive hypertension, microvascular dysfunction, and maladaptive cardio-renal crosstalk, highlighting the need to investigate adenosine signaling as a combined multi-organ target.MethodsHFpEF was induced in Dahl salt-sensitive rats by high-salt diet. Cardiac structure, function, fibrosis, oxidative stress, cytokines, renal adenosine receptors and cardiac adenosine pathway components were assessed using echocardiography, histology, proteome profiling and Western blotting. Human cardiac microvascular endothelial cells were treated with endothelin-1 in the presence of selective A2A or A2B agonists, or adenosine deaminase (ADA) inhibition, and profibrotic/oxidative stress genes were analyzed by qPCR.ResultsHypertensive rats exhibited diastolic dysfunction with preserved systolic function, cardiac and renal fibrosis, oxidative/nitrative stress, and elevated pro-inflammatory cytokines. Cardiac expression of CD39, CD73, and ADA enzymes was significantly reduced, indicating impaired adenosine metabolism, while transporters ENT2 and CNT2 were also downregulated, reflecting impairment of both equilibrative and concentrative adenosine transport. Adenosine receptor profiles were altered: A1 expression increased, A2A decreased, and A2B and A3 selectively upregulated in hypertensive, but not HFpEF, animals. In the kidney, A1 and A2A receptor expression showed region-specific, time-dependent changes. In human endothelial cells, A2A activation or ADA inhibition suppressed endothelin-1-induced COL1, COL3, and TGF-β1 expression, whereas A2B had no effect. Both A2A and A2B restored MnSOD expression, while NOX4 was selectively increased by A2B. Only A2A activation induced CAT expression, highlighting its stronger antioxidant role.ConclusionImpaired adenosine metabolism and transport, along with altered receptor signaling contribute to HFpEF progression. Selective A2A activation attenuates endothelial pro-fibrotic profiles and restores antioxidant defenses, supporting its therapeutic potential. Renal receptor changes reinforce maladaptive cardio-renal crosstalk, emphasizing the importance of multi-organ adenosine modulation in HFpEF and encouraging further translational studies.

The aryl hydrocarbon receptor (AHR) is a ligand-activated anti-inflammatory transcription factor and the first-in-class AHR-modulating topical drug, tapinarof, is approved for mild-to-moderate psoriasis. The AHR pathway is tightly regulated, including through the cytochrome P450 1 (CYP1) enzymes, which degrade AHR ligands providing negative feedback. Here we investigate the AHR pathway in depth to uncover the molecular mechanisms preventing its anti-inflammatory effect in psoriasis. Three independent cohorts of patients (n = 47) with moderate-to-severe psoriasis not receiving systemic therapy were recruited, alongside four cohorts of age-, sex- and ethnicity-matched healthy volunteers (n = 60). We measured: (i) AHR and CYP1A1 bulk and spatial mRNA and protein in skin, (ii) in vitro modulation of AHR and CYP1A1 expression in normal human epidermal keratinocytes (NHEKs) and skin epidermal sheet by psoriasis-relevant cytokines, and (iii) CYP1 enzymatic activity in psoriasis and healthy epidermal sheets. AHR expression was decreased (n = 21, P < 0.01; n = 18, P < 0.01; n = 8, P < 0.05) in all three cohorts of psoriasis lesional skin (PsL) vs. healthy skin (n = 38, n = 9, n = 8), while CYP1A1 expression was marginally reduced (P = 0.0546; n = 16). AHR mRNA predominantly localized in the basal layer in PsL (n = 5), with reduced frequency of AHR + CYP1A1 + PsL basal keratinocytes (n = 5) vs. healthy controls (n = 9, P < 0.01). In vitro culture of epidermal sheets with an AHR ligand-induced CYP1 activity in healthy skin and psoriasis nonlesional skin but not in PsL, while upregulating CYP1A1 expression in all tissue types (n = 7, Q < 0.05). Culturing normal epidermal sheets with a proinflammatory cocktail containing tumour necrosis factor reduced CYP1 activity (n = 3, P < 0.05). Finally, AHR ligation downregulated its own mRNA expression in the presence of proinflammatory cytokines (n = 3, P < 0.001), mirroring what is observed in PsL. We have identified a multilevel dysregulation of the AHR pathway in psoriasis, abrogating its anti-inflammatory effect. These findings have important implications for novel therapeutic interventions aimed at restoring the beneficial effect of AHR in skin.

A (poly)phenol enriched fraction, named Aliophen-XP, was obtained from a patented malt and hop derived formulation (Aliophen®). Here, we combined chemical profiling with in vitro and in vivo assays to evaluate its antioxidant and anti-inflammatory activity in models relevant to psoriasis. Aliophen-XP was obtained by protein/carbohydrate depletion and C18 solid phase extraction (35% acetonitrile) and characterized by HPLC UV FLD ESI Q TOF MS/MS and nano LC Q-Orbitrap MS/MS. The fraction displayed a high abundance of barley phenolamides, predominantly hordatines and their glycosylated derivatives, together with minor flavonoid C glycosides and malt derived peptides. Total (poly)phenols and antioxidant capacity were 36.30 ± 2.15 mg quercetin equivalents/g (dw) and 81.92 ± 9.06 mg Fe2+/g (dw), respectively. In HaCaT keratinocytes, Aliophen-XP was non cytotoxic across tested exposures, markedly reduced tert-butyl hydroperoxide–induced ROS, and suppressed TNFα stimulated expression of IL 1β, IL 6, and IL 8, with concordant reduction of IL 6 protein. Aliophen-XP also increased NQO1 protein levels. In C57BL/6 J mice, preventive topical application (0.8–1.6 mg mL-1, 30 min before imiquimod) attenuated psoriasiform disease, reducing ear thickening, erythema and scaling (PASI components) and lowering tissue IL 17 A levels.The findings support further development of Aliophen-XP as a topical adjunct for psoriatic skin, including therapeutic onset regimens, formulation optimization, and fractionation guided mechanistic studies to define the active principles and pathways.

Abstract The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor with anti-inflammatory and barrier functions in skin. Activated AHR induces expression of CYP1A1, which metabolizes AHR ligands providing a negative feedback mechanism controlling AHR activation. We and others have shown a dysregulation of the AHR pathway in psoriasis and atopic dermatitis (AD), but the underlying causes are ill understood. With AHR now a therapeutic target for both conditions, we sought to investigate the AHR pathway to elucidate the mechanisms preventing its anti-inflammatory function in psoriasis and AD. Skin biopsies were taken from patients with AD (n = 17) and psoriasis (n = 22) not receiving systemic treatment and healthy volunteers matched for age, sex and site (n = 31). The biopsies were used to assess (i) AHR and CYP1A1 expression in skin by quantitative reverse-­transcriptase polymerase chain reaction and (ii) CYP1 enzymatic activity in epidermal sheets by EROD (7-ethoxyresorufin O-deethylase) assay. AHR is significantly increased in lesional AD (n = 7, P < 0.05) vs. healthy skin (n = 9), while CYP1A1 shows a downward trend in lesional AD vs. healthy controls. CYP1 enzymatic activity shows no difference (P > 0.05) in basal or inducible activity in lesional AD (n = 10) vs. healthy epidermis (n = 10). In psoriasis, AHR is significantly decreased in lesional skin (n = 18, P < 0.01) vs. healthy skin (n = 9), while CYP1A1 is marginally reduced (P = 0.055). Healthy and psoriasis epidermis cultured with an AHR ligand upregulates CYP1A1 expression in all tissues (n = 7, q < 0.05). However, the induced CYP1 enzymatic activity is inhibited in lesional psoriasis skin (n = 8, q > 0.05) vs. healthy and psoriasis nonlesional epidermis (n = 8, q < 0.05). This inhibition is driven by tumour necrosis factor (TNF), shown by a reduction of CYP1 activity in healthy epidermis cultured with TNF (10 ng mL−1; n = 4, P < 0.01), and an upward trend in CYP1 activity in lesional psoriasis epidermis treated with adalimumab (10 μg mL−1, n = 4). Taking the results together, the AHR pathway is differentially dysregulated in AD and psoriasis, with implications for therapeutic targeting of the AHR pathway, as AHR-targeting treatments may result in different levels of response in these conditions.

Introduction:Psoriasis is a chronic inflammatory skin condition driven by activated epidermal keratinocytes, dermal fibroblasts, and immune cell infiltrates, which causes tissue injury. The ecto-5'-nucleotidase CD73/adenosine pathway plays a critical role in controlling inflammatory/immune responses, yet it is dysregulated in psoriasis patients. However, the expression and function of this pathway in psoriasis remain poorly explored. Methods:In this study, we investigated the regulation of CD73 in keratinocytes and examined the anti-inflammatory effects of adenosine in keratinocytes and dermal fibroblasts under psoriatic-like conditions. HaCaT cells and primary normal human epidermal keratinocytes (NHEK) were stimulated with the M5 cytokine cocktail, comprising interleukin (IL)-1α, IL-17A, IL-22, oncostatin M (OSM), and tumor necrosis factor (TNF)-α, to induce a proinflammatory phenotype. Results:M5-treated keratinocytes release IL-1β, IL-6, and IL-8, as well as the antimicrobial peptide S100A9, and exhibit activation of the signal transducer and activator of transcription (STAT), nuclear factor-kappa B (NF-κB), mitogen-activated protein kinase (MAPK), and phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathways. We demonstrated that CD73 is upregulated in inflamed keratinocytes, with OSM identified as a regulator of CD73 expression in a Janus kinase/MAPK-dependent manner. High CD73 expression in inflamed keratinocytes is associated with increased adenosine production. In M5-stimulated keratinocytes, adenosine A2A receptors (A2AR) expression is increased, whereas A2BR expression is decreased. Functional analyses revealed that an A2AR agonist, and to a lesser extent an A2BR agonist, reduced IL-8 levels in inflamed keratinocytes. Similarly, M5-treated dermal fibroblasts released IL-1β, IL-6, and IL-8, and exhibited activation of inflammatory signaling pathways. In inflamed dermal fibroblasts, both A2AR and A2BR were upregulated, and IL-8 release was mitigated by an A2AR agonist. Discussion:In conclusion, these results provide new insights into the mechanisms by which the CD73/adenosine axis can be modulated in psoriatic conditions and may guide the development of effective strategies to mitigate inflammation.

Melanoma cells express high levels of CD73 that produce extracellular immunosuppressive adenosine. Changes in the CD73 expression occur in response to tumor environmental factors, contributing to tumor phenotype plasticity and therapeutic resistance. Previously, we have observed that CD73 expression can be up-regulated on the surface of melanoma cells in response to nutritional stress. Here, we explore the mechanism by which melanoma cells release soluble CD73 under low nutrient availability and whether this might be affected by agents targeting the proto-oncogene B-Raf (BRAF). We found that starved melanoma cells can release high levels of CD73, able to convert AMP into adenosine, and this activity is abrogated by selective CD73 inhibitors, APCP or PSB-12489. The release of CD73 from melanoma cells is mediated by the matrix metalloproteinase MMP-9. Indeed, MMP-9 inhibitors significantly reduce the levels of CD73 released from the cells, while its surface levels increase. Of relevance, melanoma cells, harboring an activating BRAF mutation, upon treatment with dabrafenib or vemurafenib, show a strong reduction of CD73 cell expression and reduced levels of CD73 released into the extracellular space. Conversely, melanoma cells resistant to dabrafenib show high expression of membrane-bound CD73 and soluble CD73 released into the culture medium. In summary, our data indicate that CD73 is released from melanoma cells. The expression of CD73 is associated with response to BRAF inhibitors. Melanoma cells developing resistance to dabrafenib show increased expression of CD73, including soluble CD73 released from cells, suggesting that CD73 is involved in acquiring resistance to treatment.

Abstract Introduction and aims Adenosine is an endogenous purine nucleoside that, by binding to adenosine receptors, modulates inflammation and immune responses. Psoriasis is a chronic immune-mediated inflammatory skin disease characterized by keratinocyte hyperproliferation and abnormal differentiation. A2A and A2B adenosine receptors modulate keratinocyte proliferation and their expression was found to be altered in psoriatic epidermis. Ecto-5′-nucleotidase (CD73) is an enzyme that hydrolyses extracellular adenosine monophosphate and represents a key control point for extracellular adenosine accumulation. CD73 is involved in the regulation of local immune response and tissue barrier function. As changes in CD73 expression and activity have been associated with other autoimmune diseases, the aim of this study was to investigate CD73 involvement in skin inflammation using in vitro and in vivo approaches. Methods In vitro experiments were performed on human keratinocyte (HaCaT) cell lines stimulated with tumour necrosis factor-α, interleukin (IL)-17A, IL-6 and IL-1α (10 ng mL−1 each), and in vivo, on the mouse model of imiquimod-induced psoriasis-like inflammation on the right ear, which was also performed on CD73−/− mice and their wildtype counterpart. CD73 expression in cell lysates and ear homogenates were evaluated using Western blot analysis. Results We found that CD73 expression was significantly increased (N = 3 wells, P < 0.05) in cytokine-treated HaCaTs compared with unstimulated cells. In vivo experiments showed increased CD73 expression on psoriatic lesions (P < 0.001, N = 3). In CD73−/− mice, imiquimod-induced lesions were exacerbated compared with the development of lesions in wildtype mice as shown by ear thickness (0.190 ± 0.02 mm, knockout mice vs. 0.138 ± 0.02 mm, wildtype mice; P < 0.05, N = 5), erythema and scaling. Conclusions Our preliminary Results show involvement of CD73 in the development of psoriasis-like inflammation, suggesting that CD73 may represent a new therapeutic target in the treatment of skin diseases. Further investigations are needed to provide an in-depth study of CD73 localization in inflamed skin.

Psoriasis is an inflammatory skin disease, that can manifest as different phenotypes, however its most common form is psoriasis vulgaris (plaque psoriasis), characterized by abnormal keratinocyte proliferation, leading to characteristic histopathological signs of acanthosis, hyperkeratosis and parakeratosis.For many years, there has been a debate regarding whether keratinocyte dysfunction leads to immune system dysregulation in psoriasis or vice versa. It is now understood that epidermal hyperplasia results from immune system activation. Besides epidermal hyperplasia, psoriatic skin shows leukocyte infiltration, evident angiogenesis in the papillary dermis, characterized by tortuous, dilated capillaries, as well as oedema.There is substantial early evidence that adenosine is a key mediator of the immune response; it derives from ATP hydrolysis and accumulates into tissue in response to systemic and local stress conditions, hypoxia, metabolic stress, inflammation. Adenosine controls several cell functions by signalling through its 4 receptor subtypes, A1, A2A, A2B and A3. Evidence suggests that adenosine may play a role in psoriasis pathogenesis by controlling several immune cell functions, keratinocyte proliferation, neo-angiogenesis. Expression of adenosine receptor varies in psoriatic skin, and this can significantly impact on tissue homeostasis. Indeed, an altered adenosine receptor profile may contribute to the dysregulation observed in psoriasis, affecting immune responses and inflammatory pathways.Here, we discuss the role of adenosine in regulating the functions of the main cell populations implied in the pathogenesis of psoriasis. Furthermore, we give evidence for adenosine signalling pathway as target for therapeutic intervention in psoriasis.

Over the years, the importance of the epithelium in the assessment of allergic sensitization and development of allergic diseases has increased. Sensitization to allergens appears to be influenced by genetic and external environmental factors. However, not all subjects exposed to environmental factors that damage epithelial cells suffer from allergic diseases. On this basis, identifying the signaling pathways that characterize the different phenotypes and endotypes of allergy is of high priority for a successful personalized therapy. Ecto-5ʹ-nucleotidase/CD73 is a membrane-bound enzyme responsible for extracellular adenosine accumulation from AMP derived, in turn, from the hydrolysis of extracellular ATP. Current knowledge suggests that CD73 expression and enzymatic activity at epithelial barriers would be of fundamental importance to control the first defense against allergens, by preserving both physical and immunological epithelial barrier functions. Here, we highlight evidence for a crucial role of CD73 in features of allergic sensitization and the potential of this enzyme as prognostic marker and target of therapeutic intervention.

Tissue inflammation is a dynamic process that develops step by step, in response to an injury, to preserve tissue integrity [...]

Adenosine is an endogenous nucleoside that regulates many physiological and pathological processes. It is derived from either the intracellular or extracellular dephosphorylation of adenosine triphosphate and interacts with cell-surface G-protein-coupled receptors. Adenosine plays a substantial role in protecting against cell damage in areas of increased tissue metabolism and preventing organ dysfunction in pathological states. Targeting adenosine metabolism and receptor signaling may be an effective therapeutic approach for human diseases, including cardiovascular and central nervous system disorders, rheumatoid arthritis, asthma, renal diseases, and cancer. Several lines of evidence have shown that many drugs exert their beneficial effects by modulating adenosine signaling pathways but this knowledge urgently needs to be summarized, and most importantly, actualized. The present review collects pharmaceuticals and pharmacological or diagnostic tools that target adenosine signaling in their primary or secondary mode of action. We overviewed FDA-approved drugs as well as those currently being studied in clinical trials. Among them are already used in clinic A2A adenosine receptor modulators like istradefylline or regadenoson, but also plenty of anti-platelet, anti-inflammatory, or immunosuppressive, and anti-cancer drugs. On the other hand, we investigated dozens of specific adenosine pathway regulators that are tested in clinical trials to treat human infectious and noninfectious diseases. In conclusion, targeting purinergic signaling represents a great therapeutic challenge. The actual knowledge of the involvement of adenosinergic signaling as part of the mechanism of action of old drugs has open a path not only for drug-repurposing but also for new therapeutic strategies.

Benefits for vitamin E intake in diseases with inflammatory components have been described and related in part, to endogenously formed metabolites (long-chain metabolites, LCM). Here, we have evaluated the role of LCM in relieving asthma features. To this aim, the endogenous vitamin E metabolite α-13′-carboxychromanol (α-T-13′-COOH) that acts as potent 5-lipoxygenase inhibitor has been administered either intraperitoneally or by oral gavage to BALB/c mice sensitized by subcutaneous injection of ovalbumin (OVA). We also have taken advantage of the metabolically stable α-T-13′-COOH derivative α-amplexichromanol (α-AC). Intraperitoneal treatment with α-T-13'-COOH reduced OVA-induced airway hyperreactivity (AHR) as well as peri-bronchial inflammatory cell infiltration. α-AC was more efficacious than α-T-13'-COOH, as demonstrated by better control of AHR and in reducing subepithelial. Both compounds exerted their protective function by reducing pulmonary leukotriene C4 levels. Beneficial effects of α-AC were coupled to inhibition of the sensitization process, as indicated by a reduction of IgE plasma levels, lung mast cell infiltration and Th2 immune response. Metabololipidomics analysis revealed that α-AC raises the pulmonary levels of prostanoids, their degradation products, and 12/15-lipoxygenase metabolites. Following oral administration, the pharmacodynamically different profile in α-T-13′-COOH and α-AC was abrogated as demonstrated by a similar and improved efficacy in controlling asthma features as well as by metabololipidomics analysis. In conclusion, this study highlights a role for LCM and of vitamin E derivatives as pharmacologically active compounds that ameliorate asthmatic features and defines an important role for endogenous vitamin E metabolites in regulating immune response underlying the sensitization process.

Background: Administration of glucocorticoids might reduce mortality in patients with severe COVID-19 but have adverse cardiometabolic effects.Objectives: to investigate the effect of systemic administration of glucocorticoids on cardiovascular complications and all-cause mortality in patients hospitalised with respiratory viral infections, including COVID-19, SARS, MERS and influenza.& nbsp;Methods: We identified randomised trials published prior to July 28th, 2021. The Mantel-Haenszel random effects method and the Hartung and Knapp adjustment were used to obtain pooled estimates of treatment effect with 95% confidence intervals.& nbsp;Results: No randomised trials of glucocorticoids for SARS, MERS or influenza reported relevant outcomes. We included eleven COVID-19 randomised trials (8109 patients). Overall, compared to placebo or standard care, glucocorticoids were not associated with a reduction of in-hospital mortality (p = 0.09). In a pre-specified sub analysis, in-hospital mortality was reduced by 19% when follow-up was restricted to 14 days from randomisation (5/11 trials, 1329 patients, p = 0.02). With longer follow-up (9/11 trials, 7874 patients), administration of glucocorticoids was associated with a trend to benefit for those requiring mechanical ventilation (RR 0.86; 95% CI 0.57-1.27) but possible harm for those not receiving oxygen at randomisation (RR 1.27; 95% CI 1.00 - 1.61), an effect that was significantly different amongst subgroups (p = 0.0359). Glucocorticoids reduced the risk of worsening renal function by 37% (4/11 trials); reported rate of other cardiovascular complications was low.& nbsp;Conclusions: Administration of systemic glucocorticoids to patients hospitalised with COVID-19 does not lower mortality overall but may reduce it in those requiring respiratory support and increase it in those who do not.

Ecto-5′-nucleotidase (CD73), the ectoenzyme that together with CD39 is responsible for extracellular ATP hydrolysis and adenosine accumulation, regulates immune/inflammatory processes by controlling innate and acquired immunity cell functions. We previously demonstrated that CD73 is required for the assessment of a controlled allergic sensitization, in mice. Here, we evaluated the response to aerosolized allergen of female-sensitized mice lacking CD73 in comparison with their wild type counterpart. Results obtained show, in mice lacking CD73, the absence of airway hyperreactivity in response to an allergen challenge, paralleled by reduced airway CD23+B cells and IL4+T cells pulmonary accumulation together with reduced mast cells accumulation and degranulation. Our findings indicate CD73 as a potential therapeutic target for allergic asthma.

The paper shows a preliminary feasibility analysis of a space project both economic and structural. The project consists of launching a new business for suppling near real time connection to EO LEO satellite through a bent pipe satellite constellation. The result of business analysis highlights a good return in the next decade which is acceptable for an innovative project in space market. Nowadays the target customer groups 146 satellites which could choose among three different kinds of subscription and pay between 10 and 20 euros per minute of downlink. The design constellation is created with experts' opinion: they have been interviewed with survey and analysed with AHP, which gives high relevance to listening window width. As concerns satellite platform, quite all of satellite subsystems have a TRL 9, for the remaining it has been destinated a large budget of research and development. Due to an estimation method of power and mass, it has been possible to evaluate the size of each satellite and the required power: satellite will have a dry mass of 25 kg approximatively and require 260 Watt of power. Due to details of each subsystem, it has been realised a 3D model with Catia v5 to visualize the satellite.