Physiological role of Prion Protein in Copper homeostasis and angiogenic mechanisms of endothelial cells

Abstract The Prion Protein (PrP) is mostly known for its role in prion diseases, where its misfolding and aggregation can cause fatal neurodegenerative conditions such as the bovine spongiform encephalopathy and human Creutzfeldt–Jakob disease. Physiologically, PrP is involved in several processes including adhesion, proliferation, differentiation and angiogenesis, but the molecular mechanisms behind its role remain unclear. PrP, due to its well-described structure, is known to be able to regulate copper homeostasis; however, copper dyshomeostasis can lead to developmental defects. We investigated PrP-dependent regulation of copper homeostasis in human endothelial cells (HUVEC) using an RNA-interference protocol. PrP knockdown did not influence cell viability in silenced HUVEC (PrPKD) compared to control cells, but significantly increased PrPKD HUVEC cells sensitivity to cytotoxic copper concentrations. A reduction of PrPKD cells reductase activity and copper ions transport capacity was observed. Furthermore, PrPKD-derived spheroids exhibited altered morphogenesis and their derived cells showed a decreased vitality 24 and 48 hours after seeding. PrPKD spheroid-derived cells also showed disrupted tubulogenesis in terms of decreased coverage area, tubule length and total nodes number on matrigel, preserving unaltered VEGF receptors expression levels. Our results highlight PrP physiological role in cellular copper homeostasis and in the angiogenesis of endothelial cells

Mediterranean Cropping Systems: The Importance of Their Economic and Environmental Sustainability

The COVID-19 pandemic has drastically changed the lives of people, as well as the production and economic systems throughout the world. The flow of raw materials and products, the supply of labor and manpower, and the purchasing power have all been changed to the detriment of individual health and well-being. Such a situation requires placing even more emphasis on the search for virtuous agricultural systems compatible with the goals of economic and environmental development so clearly defined at the world level in the last decades. The present study aimed to assess the environmental and economic performance of some typical Mediterranean crops grown under different agronomical management regimes, such as strawberry, hazelnut, apricot tree, kiwifruit, peach, olive tree, and grapevine, to emphasize the importance of the mentioned issues even in the current pandemic situation. Life cycle assessment (LCA) was used to investigate the environmental profile of the studied crops, while lifecycle costing (LCC) was performed to assess and compare the economic aspects. From the environmental perspective, the hobby-organic olive systems were the most eco-friendly cropping systems, emitting 0.031 to 0.105 kg CO2eq per kg olives, while the organic hazelnut system had the greatest impact (1.001 kg of CO2eq per kg). Apricot, kiwifruit, and peach systems used N and P inputs most effectively, while strawberry systems efficiently used fossil fuels. Olive HO-2, kiwifruit, and peach cropping systems had the lowest budgets, with the costs amounted to 0.12 € kg-1 per fruit for Olive HO-2 and 0.28 € kg-1 per fruit for both kiwifruit and peach. On the contrary, organic strawberry cultivation was the most expensive (4.77 € kg-1). The variability in results due to the large differences between contexts, such as landscape, technical knowledge, and crop management, characterized the studied agricultural systems. To easily identify sustainability classes and to diminish the impact of farming practices, a considerable effort should be expended to combine LCA with LCC, C sequestration estimates, and some other useful indicators for the environmental quality evaluation

Novel bicistronic lentiviral vectors correct β-Hexosaminidase deficiency in neural and hematopoietic stem cells and progeny: implications for in vivo and ex vivo gene therapy of GM2 gangliosidosis.

Abstract The favorable outcome of in vivo and ex vivo gene therapy approaches in several Lysosomal Storage Diseases suggests that these treatment strategies might equally benefit GM2 gangliosidosis. Tay-Sachs and Sandhoff disease (the main forms of GM2 gangliosidosis) result from mutations in either the HEXA or HEXB genes encoding, respectively, the α- or β-subunits of the lysosomal β-Hexosaminidase enzyme. In physiological conditions, α- and β-subunits combine to generate β-Hexosaminidase A (HexA, αβ) and β-Hexosaminidase B (HexB, ββ). A major impairment to establishing in vivo or ex vivo gene therapy for GM2 gangliosidosis is the need to synthesize the α- and β-subunits at high levels and with the correct stoichiometric ratio, and to safely deliver the therapeutic products to all affected tissues/organs. Here, we report the generation and in vitro validation of novel bicistronic lentiviral vectors (LVs) encoding for both the murine and human codon optimized Hexa and Hexb genes. We show that these LVs drive the safe and coordinate expression of the α- and β-subunits, leading to supranormal levels of β-Hexosaminidase activity with prevalent formation of a functional HexA in SD murine neurons and glia, murine bone marrow-derived hematopoietic stem/progenitor cells (HSPCs), and human SD fibroblasts. The restoration/overexpression of β-Hexosaminidase leads to the reduction of intracellular GM2 ganglioside storage in transduced and in cross-corrected SD murine neural progeny, indicating that the transgenic enzyme is secreted and functional. Importantly, bicistronic LVs safely and efficiently transduce human neurons/glia and CD34+ HSPCs, which are target and effector cells, respectively, in prospective in vivo and ex vivo GT approaches. We anticipate that these bicistronic LVs may overcome the current requirement of two vectors co-delivering the α- or β-subunits genes. Careful assessment of the safety and therapeutic potential of these bicistronic LVs in the SD murine model will pave the way to the clinical development of LV-based gene therapy for GM2 gangliosidosis

Epithelial ovarian cancer is infiltrated by activated effector T cells co-expressing CD39, PD-1, TIM-3, CD137 and interacting with cancer cells and myeloid cells

IntroductionDespite predicted efficacy, immunotherapy in epithelial ovarian cancer (EOC) has limited clinical benefit and the prognosis of patients remains poor. There is thus a strong need for better identifying local immune dynamics and immune-suppressive pathways limiting T-cell mediated anti-tumor immunity.MethodsIn this observational study we analyzed by immunohistochemistry, gene expression profiling and flow cytometry the antigenic landscape and immune composition of 48 EOC specimens, with a focus on tumor-infiltrating lymphocytes (TILs).ResultsActivated T cells showing features of partial exhaustion with a CD137+CD39+PD-1+TIM-3+CD45RA-CD62L-CD95+ surface profile were exclusively present in EOC specimens but not in corresponding peripheral blood or ascitic fluid, indicating that the tumor microenvironment might sustain this peculiar phenotype. Interestingly, while neoplastic cells expressed several tumor-associated antigens possibly able to stimulate tumor-specific TILs, macrophages provided both co-stimulatory and inhibitory signals and were more abundant in TILs-enriched specimens harboring the CD137+CD39+PD-1+TIM-3+CD45RA-CD62L-CD95+ signature.ConclusionThese data demonstrate that EOC is enriched in CD137+CD39+PD-1+TIM-3+CD45RA-CD62L-CD95+ T lymphocytes, a phenotype possibly modulated by antigen recognition on neoplastic cells and by a combination of inhibitory and co-stimulatory signals largely provided by infiltrating myeloid cells. Furthermore, we have identified immunosuppressive pathways potentially hampering local immunity which might be targeted by immunotherapeutic approaches

Cellular metabolism and diseases

No abstract available

OMICS Sciences: toward omics personalized medicine

The omics sciences of systems biology including genomics, transcriptomics, lipidomics, metabolomics, and pro-teomics, aim at understanding the biological mechanisms that give rise to the phenotype of an organism by using high-throughput technologies with the promise of great medical advances. The importance of all these sciences is that all, with the exception of genomics are context dependent. Genome is constant in time and place in each cell of an organism, but the entire complement of messenger RNA molecules, proteins and metabolites in a cell, tissue, organ or organism varies with physiological, pathological or developmental conditions (Keusch 2006). The term "omics" represents the study of biological processes as systems. It deciphers the dynamic interactions between the numerous components of a biological system to analyse networks, pathways, and interactive rela-tions that exist among them, such as genes, transcripts, proteins, metabolites, and cells. This new scientific vision has opened the way to new research strategies and experimental technologies that have transformed the study of virtually all life processes. Expansion of the "–ome" concept was incessant and has created a host of new terms, including bacteriome, cardiome, epigenome, erythrome, immunome, microbiome, neurome, connectome, osteome, physiome, proteinome, transportome, degradome, psychome, transcriptome, and many others. In the present review, these concepts are briefly introduced with a major focus towards proteomics

Mediterranean Cropping Systems: The Importance of Their Economic and Environmental Sustainability

The COVID-19 pandemic has drastically changed the lives of people, as well as the production and economic systems throughout the world. The flow of raw materials and products, the supply of labor and manpower, and the purchasing power have all been changed to the detriment of individual health and well-being. Such a situation requires placing even more emphasis on the search for virtuous agricultural systems compatible with the goals of economic and environmental development so clearly defined at the world level in the last decades. The present study aimed to assess the environmental and economic performance of some typical Mediterranean crops grown under different agronomical management regimes, such as strawberry, hazelnut, apricot tree, kiwifruit, peach, olive tree, and grapevine, to emphasize the importance of the mentioned issues even in the current pandemic situation. Life cycle assessment (LCA) was used to investigate the environmental profile of the studied crops, while lifecycle costing (LCC) was performed to assess and compare the economic aspects. From the environmental perspective, the hobby-organic olive systems were the most eco-friendly cropping systems, emitting 0.031 to 0.105 kg CO2eq per kg olives, while the organic hazelnut system had the greatest impact (1.001 kg of CO2eq per kg). Apricot, kiwifruit, and peach systems used N and P inputs most effectively, while strawberry systems efficiently used fossil fuels. Olive HO-2, kiwifruit, and peach cropping systems had the lowest budgets, with the costs amounted to 0.12 € kg-1 per fruit for Olive HO-2 and 0.28 € kg-1 per fruit for both kiwifruit and peach. On the contrary, organic strawberry cultivation was the most expensive (4.77 € kg-1). The variability in results due to the large differences between contexts, such as landscape, technical knowledge, and crop management, characterized the studied agricultural systems. To easily identify sustainability classes and to diminish the impact of farming practices, a considerable effort should be expended to combine LCA with LCC, C sequestration estimates, and some other useful indicators for the environmental quality evaluation

Mediterranean Cropping Systems: The Importance of Their Economic and Environmental Sustainability

The COVID-19 pandemic has drastically changed the lives of people, as well as the production and economic systems throughout the world. The flow of raw materials and products, the supply of labor and manpower, and the purchasing power have all been changed to the detriment of individual health and well-being. Such a situation requires placing even more emphasis on the search for virtuous agricultural systems compatible with the goals of economic and environmental development so clearly defined at the world level in the last decades. The present study aimed to assess the environmental and economic performance of some typical Mediterranean crops grown under different agronomical management regimes, such as strawberry, hazelnut, apricot tree, kiwifruit, peach, olive tree, and grapevine, to emphasize the importance of the mentioned issues even in the current pandemic situation. Life cycle assessment (LCA) was used to investigate the environmental profile of the studied crops, while lifecycle costing (LCC) was performed to assess and compare the economic aspects. From the environmental perspective, the hobby-organic olive systems were the most eco-friendly cropping systems, emitting 0.031 to 0.105 kg CO2eq per kg olives, while the organic hazelnut system had the greatest impact (1.001 kg of CO2eq per kg). Apricot, kiwifruit, and peach systems used N and P inputs most effectively, while strawberry systems efficiently used fossil fuels. Olive HO-2, kiwifruit, and peach cropping systems had the lowest budgets, with the costs amounted to 0.12 € kg-1 per fruit for Olive HO-2 and 0.28 € kg-1 per fruit for both kiwifruit and peach. On the contrary, organic strawberry cultivation was the most expensive (4.77 € kg-1). The variability in results due to the large differences between contexts, such as landscape, technical knowledge, and crop management, characterized the studied agricultural systems. To easily identify sustainability classes and to diminish the impact of farming practices, a considerable effort should be expended to combine LCA with LCC, C sequestration estimates, and some other useful indicators for the environmental quality evaluation