Microplastic accumulation and ecological impacts on benthic invertebrates: Insights from a microcosm experiment.

Microplastic (MP) pollution poses a global concern, especially for benthic invertebrates. This one-month study investigated the accumulation of small MP polymers (polypropylene and polyester resin, 3-500 µm, 250 µg L-1) in benthic invertebrates and on one alga species. Results revealed species-specific preferences for MP size and type, driven by ingestion, adhesion, or avoidance behaviours. Polyester resin accumulated in Mytilus galloprovincialis, Chamelea gallina, Hexaplex trunculus, and Paranemonia cinerea, while polypropylene accumulated on Ulva rigida. Over time, MP accumulation decreased in count but not size, averaging 6.2 ± 5.0 particles per individual after a month. MP were mainly found inside of the organisms, especially in the gut, gills, and gonads and externally adherent MP ranged from 11 to 35 % of the total. Biochemical energy assessments after two weeks of MP exposure indicated energy gains for water column species but energy loss for sediment-associated species, highlighting the susceptibility of infaunal benthic communities to MP contamination.

Silver nitroprusside as an efficient chemodynamic therapeutic agent and a peroxynitrite nanogenerator for targeted cancer therapies.

INTRODUCTION: Chemodynamic therapy (CDT) holds great promise in achieving cancer therapy through Fenton and Fenton-like reactions, which generate highly toxic reactive species. However, CDT is limited by the lower amount of catalyst ions that can decompose already existing intracellular H2O2 and produce reactive oxygen species (ROS) to attain a therapeutic outcome. OBJECTIVES: To overcome these limitations, a tailored approach, which utilizes dual metals cations (Ag+, Fe2+) based silver pentacyanonitrosylferrate or silver nitroprusside (AgNP) were developed for Fenton like reactions that can specifically kill cancer cells by taking advantage of tumor acidic environment without used of any external stimuli. METHODS: A simple solution mixing procedure was used to synthesize AgNP as CDT agent. AgNP were structurally and morphologically characterized, and it was observed that a minimal dose of AgNP is required to destroy cancer cells with limited effects on normal cells. Moreover, comprehensive in vitro studies were conducted to evaluate antitumoral mechanism. RESULTS: AgNP have an effective ability to decompose endogenous H2O2 in cells. The decomposed endogenous H2O2 generates several different types of reactive species (•OH, O2•-) including peroxynitrite (ONOO-) species as apoptotic inducers that kill cancer cells, specifically. Cellular internalization data demonstrated that in short time, AgNP enters in lysosomes, avoid degradation and due to the acidic pH of lysosomes significantly generate high ROS levels. These data are further confirmed by the activation of different oxidative genes. Additionally, we demonstrated the biocompatibility of AgNP on mouse liver and ovarian organoids as an ex vivo model while AgNP showed the therapeutic efficacy on patient derived tumor organoids (PDTO). CONCLUSION: This work demonstrates the therapeutic application of silver nitroprusside as a multiple ROS generator utilizing Fenton like reaction. Thereby, our study exhibits a potential application of CDT against HGSOC (High Grade Serous Ovarian Cancer), a deadly cancer through altering the redox homeostasis.

Additives, plasticizers, small microplastics (<100 µm), and other microlitter components in the gastrointestinal tract of commercial teleost fish: Method of extraction, purification, quantification, and characterization using Micro-FTIR.

One of the aims of this study is the development of a pretreatment method for additives, plasticizers and other components of micro-litter (APFs), and small microplastics (SMPs <100 µm) in the gastrointestinal tract (GIT) of five of the most widely distributed and consumed commercial fish species, Engraulis encrasiculos, Sardina pilchardus, Mullus surmuletus, Solea solea, and Sparus aurata. The second aim was to develop a simultaneous quantification and identification method via Micro-FTIR of APFs and SMPs ingested by these commercial fish species. The distribution of SMPs and APFs is characteristically different for each species investigated. E. encrasiculos and S. pilchardus had a higher weight of SMPs than the other species investigated. Regarding APFs, the highest abundance was observed in E. encrasiculos. This study highlights the importance of studying additives and plasticizers that can be used as efficient proxies of microplastics, as shown by the presence of vulcanizing agents such as Vanax®.

Early Warnings by Liver Organoids on Short- and Long-Chain PFAS Toxicity.

Short-chain per-fluoroalkyl substances (PFAS) have replaced long-chains in many applications, however the toxicity and its mode of action and interactions due to the large number of these compounds and their mixtures is still poorly understood. The paper aims to compare the effects on mouse liver organoids (target organ for bioaccumulation) of two long-chain PFAS (perfluorooctane sulfonate -PFOS-, perfluorooctanoic acid -PFOA) and two short-chain PFAS commonly utilized in the industry (heptafluorobutyric acid -HFBA-, Pentafluoropropionic anhydride-PFPA) to identify the mode of action of these classes of contaminants. Cytomorphological aberrations and ALT/GDH enzyme disruption were identified but no acute toxicity endpoint neither apoptosis was detected by the two tested short-chain PFAS. After cytomorphological analysis, it is evident that short-chain PFAS affected organoid morphology inducing a reduction of cytostructural complexity and aberrant cytological features. Conversely, EC50 values of 670 ± 30 µM and 895 ± 7 µM were measured for PFOS and PFOA, respectively, together with strong ALT/GDH enzyme disruption, caspase 3 and 7 apoptosis activation and deep loss of architectural complexity of organoids in the range of 500-1000 µM. Eventually, biochemical markers and histology analysis confirmed the sensitivity of organoid tests that could be used as a fast and reproducible platform to test many PFAS and mixtures saving time and at low cost in comparison with in vivo tests. Organoids testing could be introduced as an innovative platform to assess the toxicity to fast recognize potentially dangerous pollutants.

A carrier free delivery system of a monoacylglycerol lipase hydrophobic inhibitor.

Monoacylglycerol lipase (MAGL) is an emerging therapeutic target for cancer. It is involved in lipid metabolism and its inhibition impairs many hallmarks of cancer including cell proliferation, migration/invasion and tumor growth. For these reasons, our group has recently developed a potent reversible MAGL inhibitor (MAGL23), which showed promising anticancer activities. Here in, to improve its pharmacological properties, a nanoformulation based on nanocrystals coated with albumin was prepared for therapeutic applications. MAGL23 was solubilized by a nanocrystallization method with Pluronic F-127 as surfactant into an organic solvent and was recovered as nanocrystals in water after solvent evaporation. Finally, the solubilized nanocrystals were stabilized by human serum albumin to create a smart delivery carrier. An in-silico prediction (lipophilicity, structure at different pH and solubility in water), as well as experimental studies (solubility), have been performed to check the chemical properties of the inhibitor and nanocrystals. The solubility in water increases from less than 0.01 mg/mL (0.0008 mg/mL, predicted) up to 0.82 mg/mL in water. The formulated inhibitor maintained its potency in ovarian and colon cancer cell lines as the free drug. Furthermore, the system was thoroughly observed at each step of the solubilization process till the final formulation stage by different spectroscopic techniques and a comparative study was performed to check the effects of Pluronic F-127 and CTAB as surfactants. The formulated system is favorable to release the drug at physiological pH conditions (at pH 7.4, after 24 h, less than 20% of compound is released). In vivo studies have shown that albumin-complexed nanocrystals increase the therapeutic window of MAGL23 along with a favorable biodistribution. As per our knowledge, we are reporting the first ever nanoformulation of a MAGL inhibitor, which is promising as a therapeutic system where the MAGL enzyme is involved, especially for cancer therapeutic applications.

Microplastic accumulation in commercial fish from the Adriatic Sea.

We assessed the presence and concentration of microplastics in fishes from the Northern Adriatic Sea on sale for human consumption. Microplastics were extracted from the digestive tract of 180 specimens, belonging to 6 different species (2 pelagic, and 4 demersal). Microplastics were identified through Nile red staining, and selected particles were analyzed by µ-Raman spectroscopy. The 47.8% of examined fish were found to contain microplastics (233 fragments). The mean concentration of microplastics in the 6 species ranged between 4.11 ± 2.85 items/individual in Adriatic soles, and 1.75 ± 0.71 items/individual in pilchards. Microplastics ranged in size between 0.054 mm and 0.765 mm. Polyethylene and polypropylene were the polymer types identified through µ-Raman spectroscopy. Despite our estimates are underestimations, these results are of particular concern, as the fish analyzed are consumed without being eviscerated, and the microplastics they contain are transferred directly to humans.

C-phycoerythrin from Phormidium persicinum Prevents Acute Kidney Injury by Attenuating Oxidative and Endoplasmic Reticulum Stress.

C-phycoerythrin (C-PE) is a phycobiliprotein that prevents oxidative stress and cell damage. The aim of this study was to evaluate whether C-PE also counteracts endoplasmic reticulum (ER) stress as a mechanism contributing to its nephroprotective activity. After C-PE was purified from Phormidium persicinum by using size exclusion chromatography, it was characterized by spectrometry and fluorometry. A mouse model of HgCl2-induced acute kidney injury (AKI) was used to assess the effect of C-PE treatment (at 25, 50, or 100 mg/kg of body weight) on oxidative stress, the redox environment, and renal damage. ER stress was examined with the same model and C-PE treatment at 100 mg/kg. C-PE diminished oxidative stress and cell damage in a dose-dependent manner by impeding the decrease in expression of nephrin and podocin normally caused by mercury intoxication. It reduced ER stress by preventing the activation of the inositol-requiring enzyme-1α (IRE1α) pathway and avoiding caspase-mediated cell death, while leaving the expression of protein kinase RNA-like ER kinase (PERK) and activating transcription factor 6α (ATF6α) pathways unmodified. Hence, C-PE exhibited a nephroprotective effect on HgCl2-induced AKI by reducing oxidative stress and ER stress.

Habitat heterogeneity: A confounding factor for the effect of pollutants on macrobenthic community in coastal waters.

Discrepancies were found between Ecological Status of sediments evaluated though the analysis of macrobenthic community and chemical analyses along a heavily trafficked and contaminated maritime seaway in Tyrrhenian Sea. Chemical analyses showed some sites highly contaminated by PAHs and metals with values exceeding thresholds for Good Chemical status and representing potential toxicological risk for benthic animals. Conversely, macrobenthic communities were highly rich and diverse, with Ecological Status (through M-AMBI) Good and High at each site. Sampling depth, total organic carbon, and total nitrogen were the major factors influencing biotic indices (AMBI, M-AMBI, S, H), even if concentrations of PAHs and metals (V, As, and Fe) contributed in explaining a part of indices variability. Habitat heterogeneity of sampled sites is likely acting as a confounding factor for two reasons: (1) high variability of environmental parameters leads to high richness and diversity, to which M-AMBI is sensitive, and (2) environmental parameters explained part of the variability of indices, together with contaminants. Our results suggested the importance of considering natural variability as a fundamental step of environmental impact assessment, for the correct interpretation of biotic indices.

Temporal variation of floatable plastic particles in the largest Italian river, the Po.

We investigated the temporal concentration of floatable plastic particles in the Po river, the largest contributor of freshwater to the Adriatic Sea. Surface waters were sampled in 2019 with a Manta trawl, with almost bimonthly frequency. In total, 5063 plastic particles were collected, 80.6% of which were microplastics (<5 mm). Characterization through FT-IR spectroscopy evidenced 7 polymers, of which polyethylene, polypropylene and polystyrene accounted for 40.5%, 25.7%, and 14.9%, respectively. The highest plastic concentration was recorded in December (3.47 particles m-3; 5.89 mg m-3), while the lowest in August (0.29 ± 0.01 particles m-3; 0.22 ± 0.08 mg m-3). We estimated the annual load of floatable plastic particles carried by the Po river to be 145 tons. Plastic particle load was correlated with the hydraulic regime, and was higher in autumn and spring months.

Environmental restoration by aquatic angiosperm transplants in transitional water systems: The Venice Lagoon as a case study.

The paper reports the results obtained after 4 years of aquatic angiosperm transplants in areas of the Venice Lagoon (North Adriatic Sea, Mediterranean) where meadows almost disappeared due to eutrophication, pollution and overexploitation of clam resources. The project LIFE12 NAT/IT/000331-SeResto, funded by the European Union, allowed to recolonize the Habitat 1150* (coastal lagoons) in the northernmost part of the lagoon, by extensive manual transplants of small sods or single rhizomes of Zostera marina, Zostera noltei, Ruppia cirrhosa and, in some stations also of Cymodocea nodosa. Over the 4 years of the project more than 75,000 rhizomes were transplanted in 35 stations with the support of local stakeholders (fishermen, hunters and sport clubs). Plants took root in 32 stations forming extensive meadows on a surface of approx. 10 km2 even if some failures were recorded in areas affected by outflows of freshwater rich in nutrients and suspended particulate matter. The rapid recovery of the ecological status of the involved areas was the result of this meadow restoration, which was in compliance with Water Framework Directive (WFD 2000/60/EC) objectives. Moreover, the monitoring of environmental parameters in the water column and in surface sediments allowed to identify the best conditions for successful transplants. Small, widespread interventions and the participation of local stakeholders in the environmental recovery, make this action economically cheap and easily transposable in other similar environments.

Biogenic iron-silver nanoparticles inhibit bacterial biofilm formation due to Ag+ release as determined by a novel phycoerythrin-based assay.

Silver nanoparticles (Ag-NPs) can be considered as a cost-effective alternative to antibiotics. In the presence of Fe(III)-citrate and Ag+, Klebsiella oxytoca DSM 29614 produces biogenic Ag-NPs embedded in its peculiar exopolysaccharide (EPS). K. oxytoca DSM 29614 was cultivated in a defined growth medium-containing citrate (as sole carbon source) and supplemented with Ag+ and either low or high Fe(III) concentration. As inferred from elemental analysis, transmission and scanning electron microscopy, Fourier transform infrared spectrometry and dynamic light scattering, Ag-EPS NPs were produced in both conditions and contained also Fe. The production yield of high-Fe/Ag-EPS NPs was 12 times higher than the production yield of low-Fe/Ag-EPS NPs, confirming the stimulatory effect of iron. However, relative Ag content and Ag+ ion release were higher in low-Fe/Ag-EPS NPs than in high-Fe/Ag-EPS NPs, as revealed by emission-excitation spectra by luminescent spectrometry using a novel ad hoc established phycoerythrin fluorescence-based assay. Interestingly, high and low-Fe/Ag-EPS NPs showed different and growth medium-dependent minimal inhibitory concentrations against Staphylococcus aureus ATCC 29213 and Pseudomonas aeruginosa ATCC 15442. In addition, low-Fe/Ag-EPS NPs exert inhibition of staphylococcal and pseudomonal biofilm formation, while high-Fe/Ag-EPS NPs inhibits staphylococcal biofilm formation only. Altogether, these results, highlighting the different capability of Ag+ release, support the idea that Fe/Ag-EPS NPs produced by K. oxytoca DSM 29614 can be considered as promising candidates in the development of specific antibacterial and anti-biofilm agents.Key points • Klebsiella oxytoca DSM 29614 produces bimetal nanoparticles containing Fe and Ag.• Fe concentration in growth medium affects nanoparticle yield and composition.• Phycoerythrin fluorescence-based assay was developed to determine Ag+release.• Antimicrobial efficacy of bimetal nanoparticle parallels Ag+ions release.

Microplastic accumulation in benthic invertebrates in Terra Nova Bay (Ross Sea, Antarctica).

Microplastic contamination of the benthic invertebrate fauna in Terra Nova Bay (Ross Sea, Antarctica) was determined. Twelve macrobenthic species, characterized by different feeding strategies, were selected at 3 sampling sites at increasing distance from the Italian Scientific Base (Mario Zucchelli, Camp Icarus, Adelie Cove). The 83% of the analyzed macrobenthic species contained microplastics (0.01-3.29 items mg-1). The size of the particles, measured by Feret diameter, ranged from 33 to 1000 µm with the highest relative abundance between 50 and 100 µm. Filter-feeders and grazers displayed values of microplastic contamination from 3 to 5 times higher than omnivores and predators, leading to the hypothesis that there is no evident bioaccumulation through the food web. The prevalent polymers identified by micro-FTIR were nylon (86%) and polyethylene (5%); other polymers identified in Antarctic benthos were polytetrafluoroethylene, polyoxymethylene, phenolic resin, polypropylene, polystyrene resin and XT polymer.

Hazardous effects of silver nanoparticles for primary producers in transitional water systems: The case of the seaweed Ulva rigida C. Agardh.

The acute toxicity of citrate capped silver nanoparticles (AgNP) and silver nitrate was evaluated on the marine macroalga Ulva rigida C. Agardh (1823). Silver bioaccumulation, ultrastructural chloroplast damages verified by TEM microscopy, inhibition of primary production, neutral lipid production and oxidative stress were observed after 24 h of exposure to AgNP. The toxic effects of silver nitrate in artificial seawater started from a concentration of 0.05 ppm and was more toxic than AgNP that produced effects from a concentration of 0.1 ppm. However only AgNP induced lipid peroxidation in U. rigida. The addition of natural organic and inorganic ligands, represented by transparent exopolymer particles (TEP) and clay, drastically reduced AgNP acute toxicity in a ratio AgNP:ligand of 1:100 and 1:200, respectively. The findings suggest a marked toxicity of Ag on marine macroalgae which however should be mitigated by the high natural ligand concentrations of the transitional environments.

The difficulty of disentangling natural from anthropogenic forcing factors makes the evaluation of ecological quality problematic: A case study from Adriatic lagoons.

The complex and dynamic nature of transitional ecosystems pose problems for the assessment of the Ecological Quality Status required by the European Water Framework Directive (WFD; 2000/60/EC). In six Adriatic lagoons, Ecological Quality Status was studied by comparing a biotic index based on macrophytes (MaQI), and three indices based on invertebrates (M-AMBI, M-bAMBI, and ISD). Ecological Status evaluated though MaQI and ISD resulted in quite degraded ecosystems (moderate/poor/bad), with only opportunistic algae and macrobenthic communities dominated by small size classes. Those results were supported by physico-chemical parameters, indicating high nutrients inputs, and anthropogenic pressures related with agriculture and fishery activities. Ecological Status obtained with M-AMBI and M-bAMBI was higher, with some sites reaching even the "good" status. The best response to anthropogenic pressures, in terms of a pressure index, was obtained by M-AMBI and M-bAMBI. Nevertheless, the response of used metrics (such as AMBI and bAMBI) to environmental variables not related to anthropogenic impact, and the high heterogeneity of physical-chemical conditions within lagoons, represent potential problems for the correct evaluation of Ecological Status of transitional waters. When different metrics give different responses it becomes a problem for managers who cannot easily make a decision on the remedial measures. The disagreement among indices arose because of the different response of biological elements to different stressors, and because the different indices based on macroinvertebrates focused on different aspects of the community, providing complementary information. So urge the need to find alternative approaches for a correct assessment of Ecological Status, with the combination of different biological elements, and considering the development of new indices (e.g. M-bAMBI) or refinement of the existing ones.

Spatial distribution, bioaccumulation profiles and risk for consumption of edible bivalves: a comparison among razor clam, Manila clam and cockles in the Venice Lagoon.

Despite their ecological and economical relevance, a lack of data is still occurring about the distribution, abundance, bioaccumulation and risks for consumption of some edible bivalves. The present study has been carried out in the Venice Lagoon taking into account three bivalve species to investigate i) the possible relationship among the biological features-distribution, abundance and bioaccumulation patterns- of razor clams (Solen marginatus Pulteney, 1799), Manila clams (Ruditapes philippinarum Adams and Reeve, 1850) and lagoon cockles (Cerastoderma glaucum Poiret, 1789) and the hydrological features, namely sediment physico-chemical characteristics and contamination; ii) their specific role as bioindicators of inorganic contamination; iii) the possible risks for human health associated with the consumption of these edible bivalves. Results showed that species distribution and abundance, especially for razor clams, was mainly influenced by environmental conditions and sediment granulometric composition, above metal(loid) contamination. The contamination patterns were different among species, as Manila clam generally showed higher bioaccumulation values for most of the metal(loid)s, whilst lagoon cockles preferentially accumulated Ni. Eventually, a serious concern exists for the human consumption, for all species and investigated sites regarding As. This study will raise attention on the effects of bioaccumulation of inorganic pollutants by edible bivalves and risks for consumers' safety, especially concerning razor clams and cockles, for which a critical lack of data on metal(loid) bioaccumulation occurs from the Northern Adriatic Sea.

Nitric oxide modulation of renal and cardiac hemodynamics in type 2 diabetes.

OBJECTIVE: To evaluate endothelial function in type 2 diabetic patients with and without diabetic nephropathy. METHODS: We studied the effects of systemic infusion of the nitric oxide (NO) synthase inhibitor NG-monomethyl-l-arginine (L-NMMA) on cardiovascular and renal hemodynamics in six type 2 diabetic patients with microalbuminuria (D2-MA), six type 2 diabetic patients with normoalbuminuria (D2-NA) and five control subjects. Both type 2 diabetic patients and control subjects had mild arterial hypertension. RESULTS: L-NMMA infusion decreased the cardiac index in all groups. A reduction in glomerular filtration rate (GFR) and an increase in filtration fraction were observed only in controls. Renal plasma flow decreased in controls and D2-NA patients and renal vascular resistance increased in all groups. CONCLUSIONS: The effect of L-NMMA on cardiac output was similar in controls and type 2 diabetic patients with and without diabetic nephropathy. In contrast, the effect on GFR was impaired in both diabetic groups, suggesting that glomerular NO homeostasis is altered in type 2 diabetes. Moreover the discrepancy, in diabetic patients, between cardiac and renal effects during L-NMMA infusion suggests that the modulation of glomerular hemodynamics is independent from NO-regulated cardiac output.