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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the RNA virus responsible for the coronavirus disease 2019 (COVID-19) pandemic. Although SARS-CoV-2 was reported to alter several cellular pathways, its impact on DNA integrity and the mechanisms involved remain unknown. Here we show that SARS-CoV-2 causes DNA damage and elicits an altered DNA damage response. Mechanistically, SARS-CoV-2 proteins ORF6 and NSP13 cause degradation of the DNA damage response kinase CHK1 through proteasome and autophagy, respectively. CHK1 loss leads to deoxynucleoside triphosphate (dNTP) shortage, causing impaired S-phase progression, DNA damage, pro-inflammatory pathways activation and cellular senescence. Supplementation of deoxynucleosides reduces that. Furthermore, SARS-CoV-2 N-protein impairs 53BP1 focal recruitment by interfering with damage-induced long non-coding RNAs, thus reducing DNA repair. Key observations are recapitulated in SARS-CoV-2-infected mice and patients with COVID-19. We propose that SARS-CoV-2, by boosting ribonucleoside triphosphate levels to promote its replication at the expense of dNTPs and by hijacking damage-induced long non-coding RNAs’ biology, threatens genome integrity and causes altered DNA damage response activation, induction of inflammation and cellular senescence.

Nonhealing wounds place a significant burden on both quality of life of affected patients and health systems. Skin substitutes are applied to promote the closure of nonhealing wounds, although their efficacy is limited by inadequate vascularization. The stromal vascular fraction (SVF) from the adipose tissue is a promising therapy to overcome this limitation. Despite a few successful clinical trials, its incorporation in the clinical routine has been hampered by their inconsistent results. All these studies concluded by warranting pre-clinical work aimed at both characterizing the cell types composing the SVF and shedding light on their mechanism of action. Here, we established a model of nonhealing wound, in which we applied the SVF in combination with a clinical-grade skin substitute. We purified the SVF cells from transgenic animals to trace their fate after transplantation and observed that it gave rise to a mature vascular network composed of arteries, capillaries, veins, as well as lymphatics, structurally and functionally connected with the host circulation. Then we moved to a human-in-mouse model and confirmed that SVF-derived endothelial cells formed hybrid human-mouse vessels, that were stabilized by perivascular cells. Mechanistically, SVF-derived endothelial cells engrafted and expanded, directly contributing to the formation of new vessels, while a population of fibro-adipogenic progenitors stimulated the expansion of the host vasculature in a paracrine manner. These data have important clinical implications, as they provide a steppingstone toward the reproducible and effective adoption of the SVF as a standard care for nonhealing wounds.

Bioisosteric H/F or CH

The paper presents the preliminary results of the joint HCECR-MAIKI study of the ceramic corpus found during the HCECR excavations at the Erbil Citadel and explores its relationships with the archaeological stratigraphy.

Despite the high prevalence of ischemic heart diseases worldwide, no antibody-based treatment currently exists. Starting from the evidence that a specific isoform of the Bone Morphogenetic Protein 1 (BMP1.3) is particularly elevated in both patients and animal models of myocardial infarction, here we assess whether its inhibition by a specific monoclonal antibody reduces cardiac fibrosis. We find that this treatment reduces collagen deposition and cross-linking, paralleled by enhanced cardiomyocyte survival, both in vivo and in primary cultures of cardiac cells. Mechanistically, we show that the anti-BMP1.3 monoclonal antibody inhibits Transforming Growth Factor β pathway, thus reducing myofibroblast activation and inducing cardioprotection through BMP5. Collectively, these data support the therapeutic use of anti-BMP1.3 antibodies to prevent cardiomyocyte apoptosis, reduce collagen deposition and preserve cardiac function after ischemia., Here the authors show that a monoclonal antibody against a soluble isoform of Bone Morphogenetic Protein 1 prevents cardiac cell death, reducing fibrosis and preserving cardiac function after myocardial ischemia.

Biologics are revolutionizing the treatment of chronic diseases, such as cancer and monogenic disorders, by overcoming the limits of classic therapeutic approaches using small molecules. However, the clinical use of biologics is limited for cardiovascular diseases (CVDs) , which are the primary cause of morbidity and mortality worldwide. Here, we review the state-of-the-art use of biologics for cardiac disorders and provide a framework for understanding why they still struggle to enter the field. Some limitations are common and intrinsic to all biological drugs, whereas others depend on the complexity of cardiac disease. In our opinion, delineating these struggles will be valuable in developing and accelerating the approval of a new generation of biologics for CVDs.

This review describes the advantages of adopting a molluscan complementary model, the freshwater snail Lymnaea stagnalis, to study the neural basis of learning and memory in appetitive and avoidance classical conditioning; as well as operant conditioning of its aerial respiratory and escape behaviour. We firstly explored 'what we can teach Lymnaea' by discussing a variety of sensitive, solid, easily reproducible and simple behavioural tests that have been used to uncover the memory abilities of this model system. Answering this question will allow us to open new frontiers in neuroscience and behavioural research to enhance our understanding of how the nervous system mediates learning and memory. In fact, from a translational perspective, Lymnaea and its nervous system can help to understand the neural transformation pathways from behavioural output to sensory coding in more complex systems like the mammalian brain. Moving on to the second question: 'what can Lymnaea teach us?', it is now known that Lymnaea shares important associative learning characteristics with vertebrates, including stimulus generalization, generalization of extinction and discriminative learning, opening the possibility to use snails as animal models for neuroscience translational research.

Introduction: Mammal cardiomyocytes (CMs) lose their proliferative capacity early after birth and switch to an adult phenotype, characterized by organized sarcomeres. The mechanisms that control this switch remain largely elusive. Among the changes experienced by the heart at birth is a sudden increase in mechanical load. Hypothesis: This study tests the hypothesis that variations in mechanical loading may regulate both the proliferation and the maturation of cardiomyocytes. Methods: Engineered heart tissues (EHTs) were generated using neonatal rat heart cells, with a modified protocol allowing to alter mechanical load. Unloading is obtained by reducing the distance between the silicon posts that anchor the extremities of the EHT, while overloading is induced by changing Young’s module of posts using metal braces. Adult heart unloading in vivo was achieved by heterotopically transplanting a mouse heart into the neck of syngeneic recipient mice, which were injected with EdU twice per week. Both hearts, native and transplanted, were harvested at day 30. Results: In line with our hypothesis, mechanical unloading of developed EHTs significantly increased the percentage of EdU-, Ki67- and pH3-positive CMs at 72 hours. In contrast, increasing afterload induced CM maturation, characterized by increased cross-sectional area and force of contraction, associated with reduced proliferation. Consistently, heterotopically transplanted hearts showed a significant increase in the the number of proliferating CMs in both ventricles, which were almost absent in native hearts. Conclusions: Our data indicate that mechanical unloading induce CM proliferation in both rat EHTs and an in vivo in a mouse model of heterotopic heart transplantation. These results support our initial hypothesis and indicate that mechanical loading is a master regulator of cardiomyocytes proliferation and maturation. Ongoing experiments aim at identifying the molecular mechanisms that regulate this process.

With the increased prevalence of chronic diseases, non-healing wounds place a significant burden on both the quality of life of affected patients and health systems. Skin substitutes are applied to promote the closure of non-healing wounds, although their efficacy is limited by inadequate vascularization. The stromal vascular fraction (SVF) from the adipose tissue has emerged as a promising therapy to overcome this limitation. Despite a few successful clinical trials, its incorporation in the clinical routine has been hampered by their inconsistent results. All these studies concluded by warranting pre-clinical work aimed at both characterizing the cell types composing the SVF and shedding light on their mechanism of action. Here, we established a new model of non-healing wound, in which we applied the SVF in combination with a clinical grade skin substitute. First, we purified the SVF cells from transgenic animals to trace their fate after transplantation and observed that it gave rise to a mature vascular network composed of arteries, capillaries, veins, as well as lymphatics, structurally and functionally connected with the host circulation. Then we moved to a human-in-mouse model and confirmed that SVF-derived endothelial cells formed hybrid human-mouse vessels, that were stabilized by perivascular cells of both species. Also in this case, these vessels were connected with the host circulatory system and determined a 2-fold increase in tissue perfusion. Mechanistically, SVF-derived endothelial cells engrafted and expanded, directly contributing to the formation of new vessels, while a population of fibro-adipogenic progenitors stimulated the expansion of the host vasculature in a paracrine manner. These data have important clinical implications, as they provide a steppingstone toward the reproducible and effective adoption of the SVF as a standard care for non-healing wounds.

Funding Acknowledgements Type of funding sources: None. Introduction Pathological cardiac hypertrophy is a maladaptive remodeling response, induced by multiple pathogenetic mechanisms, including genetic mutations and hypertension. Despite the high prevalence of this condition, the precise mechanisms driving its onset and progression remain elusive. Recent evidence indicates that homotypic cell fusion between cardiomyocytes occurs in the heart of lower vertebrates in response to injury, as well as in the rodent heart during the first week after birth, when cardiomyocyte size and ploidy increase. We hypothesized that homotypic fusion of cardiomyocytes could be triggered by pressure overload and contribute to cardiac hypertrophy. Methods and Results We exploited a genetic mouse model (MHC-CreER/mTmG mice) to genetically label individual cardiomyocytes by either red or green fluorescence. These mice express a tamoxifen inducible Cre recombinase (CreER) under the control of the myosin heavy chain 6 promoter (MHC). Controlled injection of tamoxifen resulted in a mosaic heart, composed by either red or green cardiomyocytes. In this genetic background, we performed transverse aortic constriction (TAC) to induce cardiac hypertrophy and analyzed cardiomyocyte size and fluorescence at one month. The systematic analysis of more than 5000 cardiomyocytes from 6 independent experiments showed a significant increase in the number of yellow cardiomyocytes after TAC compared to sham hearts, consistent with cell-cell fusion events occurred between green and red cardiomyocytes. Moreover, these yellow hypertrophic cardiomyocytes displayed a higher rate of polynucleation, with many cells bearing three or more nuclei. Conclusions and future perspective Collectively, these data point to cardiomyocyte fusion as a new mechanism responsible for to the onset of cardiac hypertrophy upon pressure overload. We cannot exclude that yellow, polynucleated cardiomyocytes derived from the heterotypic fusion between a green cardiomyocyte and other red cell types. To rule out this possibility we are currently performing a new a set of experiments in which 100% of cardiomyocytes are labelled in green. In the case of homotypic fusion, we should not observe any yellow cells upon TAC. Proving that cardiomyocyte fusion contributes to cardiac hypertrophy in response to increased pressure load and dissecting the underlying molecular mechanisms will pave the way to novel strategies to both prevent and revert this highly prevalent pathological condition.

Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): AIRC Introduction Adult mammals fail to regenerate the myocardium after ischemic injury (1). Different strategies to achieve myocardial regeneration have been considered including reactivation of progenitor cell populations, cell replacement therapies, cell reprograming and molecules able to stimulate cardiomyocyte proliferation (2). None of these approaches has been so far successful. The supply of nutrients and oxygen to the myocardium after the occlusion of a coronary artery depends on pre-existing and newly formed collateral vessels (3). The relevance of revascularization in the context of cardiac regeneration is suggested by the evidence that angiogenesis precedes cardiomyocyte proliferation in a model of neonatal heart injury (4). Our laboratory has recently demonstrated that the heart has a low angiogenetic potential (5). To what extent this is responsible for the poor regenerative capacity of the heart remains to be determined. Thus, understanding the mechanisms blocking angiogenesis in the adult heart could lead to the development of efficient strategies to promote cardiac revascularization and regeneration. Purpose Our purpose is to understand the mechanisms responsible for the low angiogenic potential of the adult mammalian heart. In particular, we designed a proteomic approach to detect differences in the composition of the perivascular extracellular matrix between the neonatal (angiogenic) with and the adult (not angiogenic) heart. Methods and Results We adopted an in vivo biotinylation strategy to label vascular extracellular proteins in vivo, followed by protein identification by mass spectrometry. A reactive derivative of biotin was systemically injected in both neonatal and adult mice (n=4) to label vascular and peri-vascular extracellular proteins. Biotinylated proteins were purified from total hearts by streptavidin-conjugated beads, eluted, and digested with trypsin. The resulting peptides were analyzed by LC-MS/MS. Among the differentially expressed proteins, we identified lumican and its receptor, integrin beta-1. This ligand-receptor pair is known to impair tube formation by endothelial cells and to interfere with both expression and activity of matrix metalloproteinases (MMPs) in vitro (6). Western blot and gene expression analysis confirmed up-regulation of lumican in the adult compared to neonatal heart, as well as a different glycosylation pattern. Lumican knock-out both ex vivo and in vivo resulted in increased vessel density. On the other hand, overexpression of lumican impaired the proliferative capacity of cardiac endothelial cells. Finally, MMP-14 activity was inhibited by adult, but not neonatal cardiac extract. Conclusions In vivo proteomic analysis identified lumican as a major contributor of the low angiogenic potential of the adult mammalian heart. These results point to lumican silencing as a promising strategy to achieve cardiac revascularization.

Funding Acknowledgements Type of funding sources: Public grant(s) – EU funding. Main funding source(s): PREFER Introduction With the increased prevalence of chronic diseases, non-healing wounds place a significant burden on the health system, with a prevalence of 2-5%, similar to the one of heart failure. They are persistent full-thickness skin lesions that affect patients suffering from vascular disorders, such as diabetes and peripheral artery disease. Skin implants and substitutes are currently applied to promote the closure of non-healing wounds. However, both approaches are poorly effective because of lack of appropriate vascularization. Purpose To promote the neo-vascularization of non-healing wounds, we use Stromal Vascular Fraction (SVF) as innovative therapeutic opportunity for wound treatment. Here, we aim to 1) characterize and demonstrate the pro-angiogenic role of SVF cells and 2) provide pre-clinical evidence of the therapeutic efficacy of the human SVF in promoting the neo-vascularization in a new mouse model of ischemic, non-healing wound. Methods To assess capacity of SVF-derived cells to improve wound revascularization, we created a new model of non-healing wound generated by wounding an ischemic limb in mice. Human and mouse SVF was purified from adipose tissue and seeded on a clinical-grade skin substitute prior to its implantation on the ischemic wound of a recipient animal. The transplantation of human SVF into NSG immunodeficient mice was verified using species-specific antibodies, while the use of genetically modified mice allowed us to trace the fate of both endothelial and non-endothelial cells upon their transplantation into syngeneic recipient animals. The function of SVF-induced vessels was assessed by systemic injection of biotinylated lectin and by Single Photon Emission Tomography (SPECT) of the treated limb. Results At day 7 the implanted mouse SVF gives rise to a widespread vascular network composed by arteries, capillaries, veins, as well as lymphatic vessels. Similarly, human SVF-derived endothelial cells formed hybrid human-mouse vessels that were stabilized by perivascular cells. At both histological and functional analysis, these vessels were connected with the host circulatory system and determined a 2-fold increase in tissue perfusion. The comparison of the activity of human SVF from different donors allowed us to disclose its dual mechanisms of action. Conclusions Here we demonstrated the efficacy of the SVF in promoting neo-vascularization of a skin substitute in a mouse model of ischemic, non-healing wounds. Its therapeutic efficacy relies on dual mechanisms of action. On the one hand, SVF-derived ECs engraft and expand, directly forming new vascular units that colonize the scaffold and extend into surrounding tissues. On the other hand, the mesenchymal progenitors stimulate the expansion of the host vasculature, which extends into the scaffold, with the eventual appearance of donor-host hybrid vessels. Collectively, these data support the use of human SVF as a powerful cell therapy to treat non-healing wounds.

Aims Cardiac ischaemia does not elicit an efficient angiogenic response. Indeed, lack of surgical revascularization upon myocardial infarction results in cardiomyocyte death, scarring, and loss of contractile function. Clinical trials aimed at inducing therapeutic revascularization through the delivery of pro-angiogenic molecules after cardiac ischaemia have invariably failed, suggesting that endothelial cells in the heart cannot mount an efficient angiogenic response. To understand why the heart is a poorly angiogenic environment, here we compare the angiogenic response of the cardiac and skeletal muscle using a lineage tracing approach to genetically label sprouting endothelial cells. Methods and results We observed that overexpression of the vascular endothelial growth factor in the skeletal muscle potently stimulated angiogenesis, resulting in the formation of a massive number of new capillaries and arterioles. In contrast, response to the same dose of the same factor in the heart was blunted and consisted in a modest increase in the number of new arterioles. By using Apelin-CreER mice to genetically label sprouting endothelial cells we observed that different pro-angiogenic stimuli activated Apelin expression in both muscle types to a similar extent, however, only in the skeletal muscle, these cells were able to sprout, form elongated vascular tubes activating Notch signalling, and became incorporated into arteries. In the heart, Apelin-positive cells transiently persisted and failed to give rise to new vessels. When we implanted cancer cells in different organs, the abortive angiogenic response in the heart resulted in a reduced expansion of the tumour mass. Conclusion Our genetic lineage tracing indicates that cardiac endothelial cells activate Apelin expression in response to pro-angiogenic stimuli but, different from those of the skeletal muscle, fail to proliferate and form mature and structured vessels. The poor angiogenic potential of the heart is associated with reduced tumour angiogenesis and growth of cancer cells.

The intestinal tract hosts the gut microbiota (GM), actively shaping health. Bile acids(BAs) are both digestive and signaling molecules acting as hormones via the activation of farnesoid X receptor (FXR). Obstruction of bile flow initiates a cascade of pathological events ultimately leading to intestinal mucosal injury. Administration of BAs in models of obstructed bile flow counteracts these detrimental effects. Objective of this study was to investigate the effects of the novel FXR agonist 3α, 7α, 11β-Trihydroxy-6α-ethyl-5β-cholan-24-oic Acid (TC-100) on intestinal mucosa integrity and cecal microbiome composition after surgical bile duct ligation (BDL), a rodent model causing bile flow obstruction. Pharmacological FXR activation was accomplished by daily oral gavage with TC-100 for 5 days. 2 days after treatment initiation, BDL was performed. BAs measurement was carried out and the 16S rDNA (V5-V6 hyper-variable regions) extracted from the cecal content was sequenced. TC-100 activates Fxr in the gut-liver axis and this translated into a significant reduction of serum and bile BA pool size with a shift to a more hydrophilic composition, while signs of intestinal mucosal damage were prevented. Firmicutes:Bacteroidota ratio progressively increased from Sham Operated (SO) mice to TC-100-treated mice. LEfSe analysis showed that Verrucomicrobia, and particularly Akkermansia muciniphila (Amuc) increasingly recognized for improving gut homeostasis and immune functions, were strongly associated to TC-100-treated mice. Intriguingly, Amuc abundance was also negatively associated to cholic acid levels. Collectively, these data indicate that intestinal FXR activation by TC-100 prevents early signs of intestinal mucosal damage by modulating BA homeostasis and GM composition.

Conventional dendritic cells (cDCs), cDC1 and cDC2, act both to initiate immunity and maintain self-tolerance. The tryptophan metabolic enzyme indoleamine 2,3-dioxygenase 1 (IDO1) is used by cDCs in maintaining tolerance, but its role in different subsets remains unclear. At homeostasis, only mature CCR7sup+/supcDC1 expressed IDO1 that was dependent on IRF8. Lipopolysaccharide treatment induced maturation and IDO1-dependent tolerogenic activity in isolated immature cDC1, but not isolated cDC2. However, both human and mouse cDC2 could induce IDO1 and acquire tolerogenic function when co-cultured with mature cDC1 through the action of cDC1-derived l-kynurenine. Accordingly, cDC1-specific inactivation of IDO1 in vivo exacerbated disease in experimental autoimmune encephalomyelitis. This study identifies a previously unrecognized metabolic communication in which IDO1-expressing cDC1 cells extend their immunoregulatory capacity to the cDC2 subset through their production of tryptophan metabolite l-kynurenine. This metabolic axis represents a potential therapeutic target in treating autoimmune demyelinating diseases.

This article presents an analysis of the new category of Africanfuturism coined by the Nigerian American writer (or Naijamerican, as she defines herself) Nnedi Okorafor in 2019, after years of questions about the limits that the category of Afrofuturism has put over the receptions of her works. Okorafor felt the urgency to open this new horizon to better insist on the importance of stories and narratives profoundly rooted in the African continent, thus abandoning the Western models and canons of science fiction and creating new ways of looking towards the far future. Through the analysis of Okorafor’s novels (Who Fears Death?, Lagoon and Binti), interviews and posts on her blog, the article explores the potentialities of Okorafor’s speculative fiction to deal with technologies, traditions, cultures, social transformations, and how these issues inform a future Africa that could possibly be an entirely new world, in which the concept of ‘West’ and ‘colonialism’ do not have any meaning.

The article, published in the first number of the new series of East & West journal edited by ISMEO, proposes an innovative methodology, based on pattern recognition techniques, for the study of the metal arrowheads, and a new typology mainly focused on materials dated between the 6th and 4th century BCE. The authors propose a first pilot study, part of a larger project still underway, in which 167 arrowheads from the sites of Persepolis, Tol-e Ajori and Susa are taken into consideration. Eac...