Polysaccharides from Wastes of Vegetable Industrial Processing: New Opportunities for Their Eco-Friendly Re-Use

Natural polysaccharides constitute a major group of biopolymers widespread in the whole vegetable kingdom. Plant polysaccharides comprise highly heterogeneous biopolymers that play diverse biological roles as structural elements, energy reserve and biological signalling. Polysaccharides from natural sources have attracted significant interest as biotechnological products, due to their commercial uses in a wide range of industrial applications. Some of them, for example, possess strong antigenic and antipathogenic activities and are successfully employed by the pharmaceutical industry for the formulation of vaccines or as a matrix for drug-delivery applications; others are utilised as food additives taking advantage of their physical-chemical properties (emulsifying power, viscoelasticity, polyelectrolyte, adherence, bio-compatibility, stabilizer, etc). Production of these biopolymers for industrial applications has both economic and environmental costs, depending on the starting materials used as feedstocks and on the chemical/thermal treatments required for their extraction. Recently, wastes from industrial processing of vegetables for food production have been proposed as potential sources of useful polysaccharides. Indeed, transformation and packaging of fruits and vegetables generates huge amounts of wastes, since only a fraction of the incoming biomass is effectively used. The management of such waste biomasses is a problem worldwide, from both economic and environmental standpoints, and therefore research is encouraged to implement new strategies for their re-use. An advantageous option is the extraction of value added chemicals, including different kinds of polysaccharides, besides chemically different species such as polyunsaturated fatty acids, natural pigments, tannins, carotenoids, antioxidants etc. Treatment of vegetable residues by means of newly and environmentally sustainable extraction techniques represents at the moment a fascinating challenge for the valorisation of agro-industrial wastes. Remarkable examples in this framework are represented by polysaccharides isolated from wastes of tomato industrial processing and of tropical fruit juices production based on granadilla, a fruit species of Passiflora, distributed mainly in the warm temperate and tropical regions of America and Africa. These polysaccharides resulted to possess useful biotechnological properties and interesting biological activities. An accurate chemical characterization demonstrated that the main biopolymer obtained from tomato wastes was an heteropolymer with a molecular weight higher than 1 ×106 Da constituted by glucose/ xylose/ galactose/ galactosamine/ glucosamine/ fucose in a relative molar ratio of 1: 0.9: 0.5: 0.4: 0.2: trace, whereas the polysaccharide isolated from waste peels of granadilla (Passiflora liguralis) fruits (with a molecular weight higher than 1 ×106 Da as well) was characterised by the presence of six different sugar residues: xylose/ glucose/ galactose/ galactosamine/ unknown component/ fucose in the relative ratios of 1:0.5:0.2:0.06:0.05:trace. The study of rheological properties showed for both biopolymers an high thermal resistance and an high viscosity, depending, in particular for granadilla waste polysaccharide, on concentration and pH, with a maximum value of 1.4  at a concentration of 3% in distilled water and a maximum value of 7.0  in citrate buffer solution. The main point of interest was represented by their suitability to produce elastic and biodegradable films potentially useful in agriculture for mulching applications in fields protection. Further studies were performed to evaluate the biological activity of these biopolymers. Tomato waste polysaccharides resulted to inhibit NF-κB activation and iNOS gene expression in J774 macrophages by preventing the reactive species production, thus suggesting a key role of these compounds in controlling oxidative stress and/or inflammation. Biological activity of polysaccharides from granadilla peels was assessed by means of brine shrimp bioassay: the isolated compound was able to strongly inhibit the cytotoxic effects produced by avarol, whose LD50 was increased of about 10-fold with respect to the control in the absence of polysaccharide. More recent studies have been focused on lemon wastes. Citrus processing produces a considerable amount of waste biomass called “lemon pomace” consisting of peels (flavedo and albedo), pulp and seeds, mainly composed by water, soluble sugars and fibres, together with other compounds such as oils, flavonoids and vitamins. Lemon wastes, generated in huge amounts by Italian industries for “Limoncello” liquor production, have been treated in order to separate useful polysaccharides. A major polysaccharide fraction was isolated and purified: analysis by means of gel filtration allowed to determine a molecular weight higher than 1 ×106 Da. The sugar components were identified after acid hydrolysis, by means of TLC and HPAE-PAD chromatography: the main components resulted to be galactose, galacturonic acid and arabinose. Further studies are now being implemented to study both rheological properties and biological activity of lemon polysaccharides. In conclusion, polysaccharides are among the most interesting biopolymers that can be recovered from vegetables: the use of wastes from industrial processing of different vegetables together with eco-friendly extraction techniques allowed the isolation of remarkable polysaccharides. Some of them showed to possess interesting rheological properties, potential biotechnological applications and finally promising bioactivities as potential anti-inflammatory agents, as tested in different biological model systems

A Project to Context Decarbonization Analysis (PCDA) to Select the Best Energy Transition Solution Fit for Funding

An Energy Transition (ET) solutions screening has been performed in a near-Europe (EU) geography (Central Asia) to assess the sustainability of investment opportunities on green technologies in line with Environmental, Social and Governance (ESG) criteria, EU climate ambition, market trends and Technology Readiness Level (TRL). Several configurations, namely a technological solution contextualized in its Life Cycle Assessment (LCA) perspective by the overall value chain, have been studied, analysed and compared through a Carbon Energy Environmental and Cost Model (CEE&C) in a Project to Context Decarbonization Analysis (PCDA). The outcome of this study allowed to select the best set of ET Solution fit for the Central Asia context and attractive for climate funding framework. 7 typical ET technologies with 3 to 6 different decarbonization scenarios for a total of 40+ configurations have been considered, ranked and tested against strategic stakeholders’ engagement criteria (industrial partner, state investments, feedstock provider, off-taker, financing institutions) and economic and financial criteria (project authorizations, feedstock guarantee, fiscal incentives, etc.). Key Performance Indicators (KPIs) considered for the ranking of configuration are Product Carbon Footprint (CTG), Greenhouse Gas (GHG) avoidance, Levelized Cost of Carbon (LCOC) by project Capital Expenditures (CAPEX) and LCOC by cost efficiency. Our proposed paper intends to show how the results of the CEE&C model with the LCOC in a PCDA study can support stakeholders and investors to implement their geographical decarbonization ambition and strategy

Melanocyte Colonization and Pigmentation of Breast Carcinoma: Description of Two Pathological Cases and Review of Literature

Colonization of breast carcinoma by non-neoplastic melanocytes of epidermal origin was first described by Azzopardi and Eusebi in 1977. We herein report two cases on the exceptional clinical and pathological features of this phenomenon in a 66-year-old and a 51-year-old patients. The pathogenesis is not fully understood, but a disrupted basement membrane and the role of tumoral growth factors are considered essential

Moving Toward a Strategy for Addressing Climate Displacement of Marine Resources: A Proof-of-Concept

Realistic predictions of climate change effects on natural resources are central to adaptation policies that try to reduce these impacts. However, most current forecasting approaches do not incorporate species-specific, process-based biological information, which limits their ability to inform actionable strategies. Mechanistic approaches, incorporating quantitative information on functional traits, can potentially predict species- and population-specific responses that result from the cumulative impacts of small-scale processes acting at the organismal level, and can be used to infer population-level dynamics and inform natural resources management. Here we present a proof-of-concept study using the European anchovy as a model species that shows how a trait-based, mechanistic species distribution model can be used to explore the vulnerability of marine species to environmental changes, producing quantitative outputs useful for informing fisheries management. We crossed scenarios of temperature and food to generate quantitative maps of selected mechanistic model outcomes (e.g., Maximum Length and Total Reproductive Output). These results highlight changing patterns of source and sink spawning areas as well as the incidence of reproductive failure. This study demonstrates that model predictions based on functional traits can reduce the degree of uncertainty when forecasting future trends of fish stocks. However, to be effective they must be based on high spatial- and temporal resolution environmental data. Such a sensitive and spatially explicit predictive approach may be used to inform more effective adaptive management strategies of resources in novel climatic conditions

The Seascape of Demersal Fish Nursery Areas in the North Mediterranean Sea, a First Step Towards the Implementation of Spatial Planning for Trawl Fisheries

The identification of nursery grounds and other essential fish habitats of exploited stocks is a key requirement for the development of spatial conservation planning aimed at reducing the adverse impact of fishing on the exploited populations and ecosystems. The reduction in juvenile mortality is particularly relevant in the Mediterranean and is considered as one of the main prerequisites for the future sustainability of trawl fisheries. The distribution of nursery areas of 11 important commercial species of demersal fish and shellfish was analysed in the European Union Mediterranean waters using time series of bottom trawl survey data with the aim of identifying the most persistent recruitment areas. A high interspecific spatial overlap between nursery areas was mainly found along the shelf break of many different sectors of the Northern Mediterranean indicating a high potential for the implementation of conservation measures. Overlap of the nursery grounds with existing spatial fisheries management measures and trawl fisheries restricted areas was also investigated. Spatial analyses revealed considerable variation depending on species and associated habitat/depth preferences with increased protection seen in coastal nurseries and minimal protection seen for deeper nurseries (e.g. Parapenaeus longirostris 6%). This is partly attributed to existing environmental policy instruments (e.g. Habitats Directive and Mediterranean Regulation EC 1967/2006) aiming at minimising impacts on coastal priority habitats such as seagrass, coralligenous and maerl beds. The new knowledge on the distribution and persistence of demersal nurseries provided in this study can support the application of spatial conservation measures, such as the designation of no-take Marine Protected Areas in EU Mediterranean waters and their inclusion in a conservation network. The establishment of no-take zones will be consistent with the objectives of the Common Fisheries Policy applying the ecosystem approach to fisheries management and with the requirements of the Marine Strategy Framework Directive to maintain or achieve seafloor integrity and good environmental status.Versión del editor4,411

Genetic determinants in a critical domain of ns5a correlate with hepatocellular carcinoma in cirrhotic patients infected with hcv genotype 1b

HCV is an important cause of hepatocellular carcinoma (HCC). HCV NS5A domain‐1 interacts with cellular proteins inducing pro‐oncogenic pathways. Thus, we explore genetic variations in NS5A domain‐1 and their association with HCC, by analyzing 188 NS5A sequences from HCV genotype‐1b infected DAA‐naïve cirrhotic patients: 34 with HCC and 154 without HCC. Specific NS5A mutations significantly correlate with HCC: S3T (8.8% vs. 1.3%, p = 0.01), T122M (8.8% vs. 0.0%, p < 0.001), M133I (20.6% vs. 3.9%, p < 0.001), and Q181E (11.8% vs. 0.6%, p < 0.001). By multivariable analysis, the presence of >1 of them independently correlates with HCC (OR (95%CI): 21.8 (5.7–82.3); p < 0.001). Focusing on HCC‐group, the presence of these mutations correlates with higher viremia (median (IQR): 5.7 (5.4–6.2) log IU/mL vs. 5.3 (4.4–5.6) log IU/mL, p = 0.02) and lower ALT (35 (30–71) vs. 83 (48–108) U/L, p = 0.004), suggesting a role in enhancing viral fitness without affecting necroinflammation. Notably, these mutations reside in NS5A regions known to interact with cellular proteins crucial for cell‐cycle regulation (p53, p85‐PIK3, and β‐ catenin), and introduce additional phosphorylation sites, a phenomenon known to ameliorate NS5A interaction with cellular proteins. Overall, these results provide a focus for further investigations on molecular bases of HCV‐mediated oncogenesis. The role of these NS5A domain‐1 mutations in triggering pro‐oncogenic stimuli that can persist also despite achievement of sustained virological response deserves further investigation

An explainable model of host genetic interactions linked to COVID-19 severity

We employed a multifaceted computational strategy to identify the genetic factors contributing to increased risk of severe COVID-19 infection from a Whole Exome Sequencing (WES) dataset of a cohort of 2000 Italian patients. We coupled a stratified k-fold screening, to rank variants more associated with severity, with the training of multiple supervised classifiers, to predict severity based on screened features. Feature importance analysis from tree-based models allowed us to identify 16 variants with the highest support which, together with age and gender covariates, were found to be most predictive of COVID-19 severity. When tested on a follow-up cohort, our ensemble of models predicted severity with high accuracy (ACC = 81.88%; AUCROC = 96%; MCC = 61.55%). Our model recapitulated a vast literature of emerging molecular mechanisms and genetic factors linked to COVID-19 response and extends previous landmark Genome-Wide Association Studies (GWAS). It revealed a network of interplaying genetic signatures converging on established immune system and inflammatory processes linked to viral infection response. It also identified additional processes cross-talking with immune pathways, such as GPCR signaling, which might offer additional opportunities for therapeutic intervention and patient stratification. Publicly available PheWAS datasets revealed that several variants were significantly associated with phenotypic traits such as "Respiratory or thoracic disease", supporting their link with COVID-19 severity outcome.A multifaceted computational strategy identifies 16 genetic variants contributing to increased risk of severe COVID-19 infection from a Whole Exome Sequencing dataset of a cohort of Italian patients

Carriers of ADAMTS13 Rare Variants Are at High Risk of Life-Threatening COVID-19

Thrombosis of small and large vessels is reported as a key player in COVID-19 severity. However, host genetic determinants of this susceptibility are still unclear. Congenital Thrombotic Thrombocytopenic Purpura is a severe autosomal recessive disorder characterized by uncleaved ultra-large vWF and thrombotic microangiopathy, frequently triggered by infections. Carriers are reported to be asymptomatic. Exome analysis of about 3000 SARS-CoV-2 infected subjects of different severities, belonging to the GEN-COVID cohort, revealed the specific role of vWF cleaving enzyme ADAMTS13 (A disintegrin-like and metalloprotease with thrombospondin type 1 motif, 13). We report here that ultra-rare variants in a heterozygous state lead to a rare form of COVID-19 characterized by hyper-inflammation signs, which segregates in families as an autosomal dominant disorder conditioned by SARS-CoV-2 infection, sex, and age. This has clinical relevance due to the availability of drugs such as Caplacizumab, which inhibits vWF-platelet interaction, and Crizanlizumab, which, by inhibiting P-selectin binding to its ligands, prevents leukocyte recruitment and platelet aggregation at the site of vascular damage

Gain- and Loss-of-Function CFTR Alleles Are Associated with COVID-19 Clinical Outcomes

Carriers of single pathogenic variants of the CFTR (cystic fibrosis transmembrane conductance regulator) gene have a higher risk of severe COVID-19 and 14-day death. The machine learning post-Mendelian model pinpointed CFTR as a bidirectional modulator of COVID-19 outcomes. Here, we demonstrate that the rare complex allele [G576V;R668C] is associated with a milder disease via a gain-of-function mechanism. Conversely, CFTR ultra-rare alleles with reduced function are associated with disease severity either alone (dominant disorder) or with another hypomorphic allele in the second chromosome (recessive disorder) with a global residual CFTR activity between 50 to 91%. Furthermore, we characterized novel CFTR complex alleles, including [A238V;F508del], [R74W;D1270N;V201M], [I1027T;F508del], [I506V;D1168G], and simple alleles, including R347C, F1052V, Y625N, I328V, K68E, A309D, A252T, G542*, V562I, R1066H, I506V, I807M, which lead to a reduced CFTR function and thus, to more severe COVID-19. In conclusion, CFTR genetic analysis is an important tool in identifying patients at risk of severe COVID-19

The polymorphism L412F in TLR3 inhibits autophagy and is a marker of severe COVID-19 in males

The polymorphism L412F in TLR3 has been associated with several infectious diseases. However, the mechanism underlying this association is still unexplored. Here, we show that the L412F polymorphism in TLR3 is a marker of severity in COVID-19. This association increases in the sub-cohort of males. Impaired macroautophagy/autophagy and reduced TNF/TNFα production was demonstrated in HEK293 cells transfected with TLR3L412F-encoding plasmid and stimulated with specific agonist poly(I:C). A statistically significant reduced survival at 28 days was shown in L412F COVID-19 patients treated with the autophagy-inhibitor hydroxychloroquine (p = 0.038). An increased frequency of autoimmune disorders such as co-morbidity was found in L412F COVID-19 males with specific class II HLA haplotypes prone to autoantigen presentation. Our analyses indicate that L412F polymorphism makes males at risk of severe COVID-19 and provides a rationale for reinterpreting clinical trials considering autophagy pathways. Abbreviations: AP: autophagosome; AUC: area under the curve; BafA1: bafilomycin A1; COVID-19: coronavirus disease-2019; HCQ: hydroxychloroquine; RAP: rapamycin; ROC: receiver operating characteristic; SARS-CoV-2: severe acute respiratory syndrome coronavirus 2; TLR: toll like receptor; TNF/TNF-α: tumor necrosis factor