Feline herpesvirus 1 (FHV-1) enters the cell by receptor-mediated endocytosis.

Feline herpesvirus type 1 (FHV-1) is an enveloped dsDNA virus belonging to the Herpesviridae family and is considered one of the two primary viral etiological factors of feline upper respiratory tract disease. In this study, we investigated the entry of FHV-1 into host cells using two models: the AK-D cell line and primary feline skin fibroblasts (FSFs). We employed confocal microscopy, siRNA silencing, and selective inhibitors of various entry pathways. Our observations revealed that the virus enters cells via pH and dynamin-dependent endocytosis, as the infection was significantly inhibited by NH4Cl, bafilomycin A1, dynasore, and mitmab. Additionally, genistein, nystatin, and filipin treatments, siRNA knock-down of caveolin-1, as well as FHV-1 and caveolin-1 colocalization suggest the involvement of caveolin-mediated endocytosis during the entry process. siRNA knock-down of clathrin heavy chain and analysis of virus particle colocalization with clathrin indicated that clathrin-mediated endocytosis also takes part in the primary cells. This is the first study to systematically examine FHV-1 entry into host cells, and for the first time, we describe FHV-1 replication in AK-D and FSFs. IMPORTANCE Feline herpesvirus 1 (FHV-1) is one of the most prevalent viruses in cats, causing feline viral rhinotracheitis, which is responsible for over half of viral upper respiratory diseases in cats and can lead to ocular lesions resulting in loss of sight. Although the available vaccine reduces the severity of the disease, it does not prevent infection or limit virus shedding. Despite the clinical relevance, the entry mechanisms of FHV-1 have not been thoroughly studied. Considering the limitations of commonly used models based on immortalized cells, we sought to verify our findings using primary feline skin fibroblasts, the natural target for infection in cats.

The elastography of distal ulnar nerve branches in cyclists.

INTRODUCTION: Forced elbow flexion and pressure during bicycling result in ulnar nerve traction and pressure exerted in Guyon's canal or the nerve's distal branches. The compression of the nerves causes a change in their stiffness related to edema and eventually gradual fibrosis. PURPOSE: This study aimed to evaluate the elastography of terminal branches of the ulnar nerve in cyclists. STUDY DESIGN: Cross-sectional study. METHODS: Thirty cyclists, 32 healthy individuals, and 32 volunteers with ulnar nerve entrapment neuropathies participated in the study. Each participant underwent a nerve examination of the cubital tunnel, Guyon's canal and the deep and superficial branches of the ulnar nerve using shear wave elastography. The cyclist group was tested before and after a 2-hour-long workout. RESULTS: Before cycling workouts, the ulnar nerve stiffness in the cubital tunnel and Guyon's canal remained below pathological estimates. Cycling workouts altered nerve stiffness in the cubital tunnel only. Notably, the stiffness of the ulnar terminal branches in cyclists was increased even before training. The mean deep branch stiffness was 50.85 ± 7.60 kPa versus 20.43 ± 5.95 kPa (p < 0.001) in the cyclist and healthy groups, respectively, and the mean superficial branch stiffness was 44 ± 12.45 kPa versus 24.55 ± 8.05 kPa (p < 0.001), respectively. Cycling contributed to a further shift in all observed values. DISCUSSION: These observations indicate the existence of persistent anatomical changes in the distal ulnar branches in resting cyclists that result in increased stiffness of these nerves. The severity of these changes remains, however, to be determined. CONCLUSIONS: These data show elastography values of the ulnar terminal branches in healthy individuals and cyclists where despite lack of clinical symptoms that they seem to be elevated twice above the healthy range.

Elastography of nerves in the wrists of cyclists.

Purpose: This study aimed to investigate changes in the elasticity of the median and ulnar nerves in cyclists. Material and methods: The study included 30 cyclists and 2 non-biking reference groups that included 32 healthy volunteers and 32 individuals with ulnar nerve entrapment neuropathies. All participants underwent physical, ultrasonographic, and elastographic examinations including assessment of nerve cross-sectional area (CSA) and stiffness (SWE). The cyclists' group was tested before and after a 2-hour workout. Results: The values of ulnar nerve CSA and stiffness in Guyon's canal in resting cyclists were 5.30 ± 1.51 mm2 and 49.05 ± 11.18 kPa, respectively. These values were significantly higher than in the healthy volunteers, but not higher than in the nerve entrapment group. Median nerve CSA and stiffness at rest were 9.10 ± 2.61 mm2 and 38.54 ± 14.87 kPa, respectively. Both values were higher than respective values in the healthy group. Cycling induced an increase in all these parameters, although the increase in nerve stiffness was more noticeable than in CSA. Conclusions: The elasticity of the median and ulnar nerve in cyclists remains within normal limits, questioning the belief that cyclists are at risk of nerve palsy in Guyon's canal. However, cycling workout does exert compression, resulting in transient oedema of both nerves. The dynamics of changes was more noticeable in SWE examination than in conventional ultrasound, which may depend on SWE sensitivity.

Graphene Modification by Curcuminoids as an Effective Method to Improve the Dispersion and Stability of PVC/Graphene Nanocomposites.

A large amount of graphene-related research is its use as a filler for polymer composites, including thin nanocomposite films. However, its use is limited by the need for large-scale methods to obtain high-quality filler, as well as its poor dispersion in the polymer matrix. This work presents polymer thin-film composites based on poly(vinyl chloride) (PVC) and graphene, whose surfaces were modified by curcuminoids. TGA, UV-vis, Raman spectroscopy, XPS, TEM, and SEM methods have confirmed the effectiveness of the graphene modification due to π-π interactions. The dispersion of graphene in the PVC solution was investigated by the turbidimetric method. SEM, AFM, and Raman spectroscopy methods evaluated the thin-film composite's structure. The research showed significant improvements in terms of graphene's dispersion (in solutions and PVC composites) following the application of curcuminoids. The best results were obtained for materials modified with compounds obtained from the extraction of the rhizome of Curcuma longa L. Modification of the graphene's surface with these compounds also increased the thermal and chemical stability of PVC/graphene nanocomposites.

HTCC as a Polymeric Inhibitor of SARS-CoV-2 and MERS-CoV.

Among seven coronaviruses that infect humans, three (severe acute respiratory syndrome coronavirus [SARS-CoV], Middle East respiratory syndrome coronavirus [MERS-CoV], and the newly identified severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2]) are associated with a severe, life-threatening respiratory infection and multiorgan failure. We previously proposed that the cationically modified chitosan N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride (HTCC) is a potent inhibitor of human coronavirus NL63 (HCoV-NL63). Next, we demonstrated the broad-spectrum antiviral activity of the compound, as it inhibited all low-pathogenicity human coronaviruses (HCoV-NL63, HCoV-229E, HCoV-OC43, and HCoV-HKU1). Here, using in vitro and ex vivo models of human airway epithelia, we show that HTCC effectively blocks MERS-CoV and SARS-CoV-2 infection. We also confirmed the mechanism of action for these two viruses, showing that the polymer blocks the virus entry into the host cell by interaction with the S protein.IMPORTANCE The beginning of 2020 brought us information about the novel coronavirus emerging in China. Rapid research resulted in the characterization of the pathogen, which appeared to be a member of the SARS-like cluster, commonly seen in bats. Despite the global and local efforts, the virus escaped the health care measures and rapidly spread in China and later globally, officially causing a pandemic and global crisis in March 2020. At present, different scenarios are being written to contain the virus, but the development of novel anticoronavirals for all highly pathogenic coronaviruses remains the major challenge. Here, we describe the antiviral activity of an HTCC compound, previously developed by us, which may be used as a potential inhibitor of currently circulating highly pathogenic coronaviruses-SARS-CoV-2 and MERS-CoV.

The SARS-CoV-2 ORF10 is not essential in vitro or in vivo in humans.

SARS-CoV-2 genome annotation revealed the presence of 10 open reading frames (ORFs), of which the last one (ORF10) is positioned downstream of the N gene. It is a hypothetical gene, which was speculated to encode a 38 aa protein. This hypothetical protein does not share sequence similarity with any other known protein and cannot be associated with a function. While the role of this ORF10 was proposed, there is growing evidence showing that the ORF10 is not a coding region. Here, we identified SARS-CoV-2 variants in which the ORF10 gene was prematurely terminated. The disease was not attenuated, and the transmissibility between humans was maintained. Also, in vitro, the strains replicated similarly to the related viruses with the intact ORF10. Altogether, based on clinical observation and laboratory analyses, it appears that the ORF10 protein is not essential in humans. This observation further proves that the ORF10 should not be treated as the protein-coding gene, and the genome annotations should be amended.

Replication of Severe Acute Respiratory Syndrome Coronavirus 2 in Human Respiratory Epithelium.

Currently, there are four seasonal coronaviruses associated with relatively mild respiratory tract disease in humans. However, there is also a plethora of animal coronaviruses which have the potential to cross the species border. This regularly results in the emergence of new viruses in humans. In 2002, severe acute respiratory syndrome coronavirus (SARS-CoV) emerged and rapidly disappeared in May 2003. In 2012, Middle East respiratory syndrome coronavirus (MERS-CoV) was identified as a possible threat to humans, but its pandemic potential so far is minimal, as human-to-human transmission is ineffective. The end of 2019 brought us information about severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emergence, and the virus rapidly spread in 2020, causing an unprecedented pandemic. At present, studies on the virus are carried out using a surrogate system based on the immortalized simian Vero E6 cell line. This model is convenient for diagnostics, but it has serious limitations and does not allow for understanding of the biology and evolution of the virus. Here, we show that fully differentiated human airway epithelium cultures constitute an excellent model to study infection with the novel human coronavirus SARS-CoV-2. We observed efficient replication of the virus in the tissue, with maximal replication at 2 days postinfection. The virus replicated in ciliated cells and was released apically.IMPORTANCE Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged by the end of 2019 and rapidly spread in 2020. At present, it is of utmost importance to understand the biology of the virus, rapidly assess the treatment potential of existing drugs, and develop new active compounds. While some animal models for such studies are under development, most of the research is carried out in Vero E6 cells. Here, we propose fully differentiated human airway epithelium cultures as a model for studies on SARS-CoV-2.

MASS SPECTROMETRY IN VIROLOGICAL SCIENCES.

Virology, as a branch of the life sciences, discovered mass spectrometry (MS) to be the pivotal tool around two decades ago. The technique unveiled the complex network of interactions between the living world of pro- and eukaryotes and viruses, which delivered "a piece of bad news wrapped in protein" as defined by Peter Medawar, Nobel Prize Laureate, in 1960. However, MS is constantly evolving, and novel approaches allow for a better understanding of interactions in this micro- and nanoworld. Currently, we can investigate the interplay between the virus and the cell by analyzing proteomes, interactomes, virus-cell interactions, and search for the compounds that build viral structures. In addition, by using MS, it is possible to look at the cell from the broader perspective and determine the role of viral infection on the scale of the organism, for example, monitoring the crosstalk between infected tissues and the immune system. In such a way, MS became one of the major tools for the modern virology, allowing us to see the infection in the context of the whole cell or the organism. © 2019 John Wiley & Sons Ltd. Mass Spec Rev.

Determination of the Influence of Mechanical Properties of Capsules and Seeds on the Susceptibility to Feeding of Mononychus pubctumalbum in Endangered Plant Species Iris aphylla L. and Iris sibirica L.

The aim of the study was to determine the correlation between the mechanical resistance of iris seed capsules and seeds to Mononychus punctumalbum foraging. The principal component analysis (PCA) demonstrated that the first main component referred to the variety type in 68%, and the second main component described the stage of the ontogenetic development of the plant in 26%. As indicated by the values of each parameter measured, all the parameters were found to exert a strong impact on the variability of the analyzed system. The occurrence of weevil infestation was also strongly but negatively correlated with seed wall thickness and capsule wall thickness. There was a correlation of seed max load and seed mass with the occurrence of the weevil. The analysis of the mechanical resistance of iris seed capsules (in June 9.28 N and September 6.27 N for I. sibirica and in June 6.59 N and September 2.94 N for I. aphylla) and seeds (in June 15.97 N and September 344.90 N for I. sibirica and in June 16.60 N and September 174.46 N for I. aphylla) showed significant differences between the terms and species. The PCA analysis revealed that the first variable was correlated with the occurrence of weevil foraging.

Seleno-Functionalization of Quercetin Improves the Non-Covalent Inhibition of Mpro and Its Antiviral Activity in Cells against SARS-CoV-2.

The development of new antiviral drugs against SARS-CoV-2 is a valuable long-term strategy to protect the global population from the COVID-19 pandemic complementary to the vaccination. Considering this, the viral main protease (Mpro) is among the most promising molecular targets in light of its importance during the viral replication cycle. The natural flavonoid quercetin 1 has been recently reported to be a potent Mpro inhibitor in vitro, and we explored the effect produced by the introduction of organoselenium functionalities in this scaffold. In particular, we report here a new synthetic method to prepare previously inaccessible C-8 seleno-quercetin derivatives. By screening a small library of flavonols and flavone derivatives, we observed that some compounds inhibit the protease activity in vitro. For the first time, we demonstrate that quercetin (1) and 8-(p-tolylselenyl)quercetin (2d) block SARS-CoV-2 replication in infected cells at non-toxic concentrations, with an IC50 of 192 µM and 8 µM, respectively. Based on docking experiments driven by experimental evidence, we propose a non-covalent mechanism for Mpro inhibition in which a hydrogen bond between the selenium atom and Gln189 residue in the catalytic pocket could explain the higher Mpro activity of 2d and, as a result, its better antiviral profile.

Mass Spectrometry versus Conventional Techniques of Protein Detection: Zika Virus NS3 Protease Activity towards Cellular Proteins.

Mass spectrometry (MS) used in proteomic approaches is able to detect hundreds of proteins in a single assay. Although undeniable high analytical power of MS, data acquired sometimes lead to confusing results, especially during a search of very selective, unique interactions in complex biological matrices. Here, we would like to show an example of such confusing data, providing an extensive discussion on the observed phenomenon. Our investigations focus on the interaction between the Zika virus NS3 protease, which is essential for virus replication. This enzyme is known for helping to remodel the microenvironment of the infected cells. Several reports show that this protease can process cellular substrates and thereby modify cellular pathways that are important for the virus. Herein, we explored some of the targets of NS3, clearly shown by proteomic techniques, as processed during infection. Unfortunately, we could not confirm the biological relevance of protein targets for viral infections detected by MS. Thus, although mass spectrometry is highly sensitive and useful in many instances, also being able to show directions where cell/virus interaction occurs, we believe that deep recognition of their biological role is essential to receive complete insight into the investigated process.

l-Arginine Improves Solubility and ANTI SARS-CoV-2 Mpro Activity of Rutin but Not the Antiviral Activity in Cells.

The COVID-19 pandemic outbreak prompts an urgent need for efficient therapeutics, and repurposing of known drugs has been extensively used in an attempt to get to anti-SARS-CoV-2 agents in the shortest possible time. The glycoside rutin shows manifold pharmacological activities and, despite its use being limited by its poor solubility in water, it is the active principle of many pharmaceutical preparations. We herein report our in silico and experimental investigations of rutin as a SARS-CoV-2 Mpro inhibitor and of its water solubility improvement obtained by mixing it with l-arginine. Tests of the rutin/l-arginine mixture in a cellular model of SARS-CoV-2 infection highlighted that the mixture still suffers from unfavorable pharmacokinetic properties, but nonetheless, the results of this study suggest that rutin might be a good starting point for hit optimization.

Membrane Protein of Human Coronavirus NL63 Is Responsible for Interaction with the Adhesion Receptor.

Human coronavirus NL63 (HCoV-NL63) is a common respiratory virus that causes moderately severe infections. We have previously shown that the virus uses heparan sulfate proteoglycans (HSPGs) as the initial attachment factors, facilitating viral entry into the cell. In the present study, we show that the membrane protein (M) of HCoV-NL63 mediates this attachment. Using viruslike particles lacking the spike (S) protein, we demonstrate that binding to the cell is not S protein dependent. Furthermore, we mapped the M protein site responsible for the interaction with HSPG and confirmed its relevance using a viable virus. Importantly, in silico analysis of the region responsible for HSPG binding in different clinical isolates and the Amsterdam I strain did not exhibit any signs of cell culture adaptation.IMPORTANCE It is generally accepted that the coronaviral S protein is responsible for viral interaction with a cellular receptor. Here we show that the M protein is also an important player during early stages of HCoV-NL63 infection and that the concerted action of the two proteins (M and S) is a prerequisite for effective infection. We believe that this study broadens the understanding of HCoV-NL63 biology and may also alter the way in which we perceive the first steps of cell infection with the virus. The data presented here may also be important for future research into vaccine or drug development.

Start-up of a full-scale partial nitritation-anammox MBBR without inoculum at Klagshamn WWTP.

Partial nitritation and anaerobic ammonium oxidation (PNA) is a useful process for the treatment of nitrogen-rich centrate from the dewatering of anaerobically digested sludge. A one-stage PNA moving bed biofilm reactor (MBBR) was started up without inoculum at Klagshamn wastewater treatment plant, southern Sweden. The reactor was designed to treat up to 200 kgN d-1, and heated dilution water was used during start-up. The nitrogen removal was >80% after 111 days of operation, and the nitrogen removal rate reached 1.8 gN m-2 d1 at 35 °C. The start-up period of the reactor was comparable to that of inoculated full-scale systems. The operating conditions of the system were found to be important, and online control of the free ammonia concentration played a crucial role. Ex situ batch activity tests were performed to evaluate process performance.

Ultrasound Elastography for Determination of the Age of Venous Thrombi. Evaluation of Thrombus Evolution in Patients After Sclerotherapy.

BACKGROUND: Venous thrombosis is a multicausal disease involving intravenous clot formation. It may occur spontaneously or after provoking events, such as traumatic injuries to the pelvis, upper and lower extermities, immobilization, intravascular procedures, including venous catheterization or injection. Color Doppler ultrasonography is a rapid and non-invasive technique for evaluation of venous disease. It is a very sensitive method for detection of thrombi, but has some limitations, e.g. inability to distinguish between an old and fresh thrombus. Elastography as a method for evaluation of tissue elasticity and allows more accurate assessment of venous thombosis. The thrombus solidifies significantly during the process of remodeling, thereby changing elasticity, while the tissues located around the vessel remain unchanged. Moreover, the homogeneity of the thrombus is also changed. These factors allow for exact determination of the age of the thrombus depending on changes of its elasticity. MATERIAL/METHODS: The object of this study was to assess thrombus age in patients with saphenous vein insufficiency treated with sclerotherapy. We examined 34 patients, 30 women and 4 men, aged 18-62 years. All short-listed patients with initial 7-8 mm vessel diameter underwent treatment with 3% Aetoxysklerol mixed with CO2 and other respiratory gases. Data acquisition was performed using an Esaote MyLab Twice device with LA523 linear probe. RESULTS: On the sixth day after the treatment 31 patients presented uniform, hypoechogenic thrombus in B-mode image. Ultrasound-based elasticity images (elastography) showed significant predominant red and green areas. Blue area was either invisible or appeared in insignificant grade on examined area. The mosaic image of colors appeared on 2 patients. On the fourteenth day of study 21 patients still presented hypoechogenic thrombus, whereas mixed echogenicity of thrombus appeared on 11 patients. On 28 examined patients mosaics of red, green and blue colors were observed, with blue color significantly predominating over red color. 3 patients presented mosaics of colors without a predominate color. CONCLUSIONS: Elastography as a method of thrombus evaluation, provides information about relative shrinkage of blood clot. It seems that elasatography can improve accurate assessment of the exact age of thrombus.

Cine dyscontractility index: A novel marker of mechanical dyssynchrony that predicts response to cardiac resynchronization therapy.

PURPOSE: To investigate whether magnetic resonance imaging (MRI) cine-derived dyssynchrony indices provide additional information compared to conventional tagged MRI (tMRI) acquisitions in heart failure patients undergoing cardiac resynchronization therapy (CRT). MATERIALS AND METHODS: Patients scheduled for CRT (n = 52) underwent preprocedure MRI including cine and tMRI acquisitions. Segmental strain curves were calculated for both cine and tMRI to produce a range of standard indices for direct comparison between modalities. We also proposed and evaluated a novel index of "dyscontractility," which detects the presence of focal areas with paradoxically positive circumferential strain. RESULTS: Across conventional strain indices, there was only moderate-to-poor (R = 0.3-0.6) correlation between modalities; eight cine-derived indices showed statistically significant (P < 0.05) relations to CRT outcome compared to just two tMRI-based counterparts. The novel dyscontractility index calculated on basal slice cine images (cine dyscontractility index, "CDI") was the single best predictor of clinical response to CRT (area under the curve AUC = 0.81, P < 0.001). While poorly correlated to its tMRI counterpart (R = 0.33), CDI performed significantly better in predicting response to CRT (P < 0.005), and was also numerically better than all other tMRI indices (AUC 0.53-0.76, all P for AUC comparisons <0.17). CONCLUSION: Cine-derived strain indices offer potentially new information compared to tMRI. Specifically, the novel CDI is most strongly linked to response to cardiac resynchronization therapy in a contemporary patient cohort. It utilizes readily available MRI data, is relatively straightforward to process, and compares favorably with any conventional tagging index. J. Magn. Reson. Imaging 2016;44:1483-1492.

Trodusquemine (MSI-1436) Restores Metabolic Flexibility and Mitochondrial Dynamics in Insulin-Resistant Equine Hepatic Progenitor Cells (HPCs).

Equine metabolic syndrome (EMS) is a significant global health concern in veterinary medicine. There is increasing interest in utilizing molecular agents to modulate hepatocyte function for potential clinical applications. Recent studies have shown promising results in inhibiting protein tyrosine phosphatase (PTP1B) to maintain cell function in various models. In this study, we investigated the effects of the inhibitor Trodusquemine (MSI-1436) on equine hepatic progenitor cells (HPCs) under lipotoxic conditions. We examined proliferative activity, glucose uptake, and mitochondrial morphogenesis. Our study found that MSI-1436 promotes HPC entry into the cell cycle and protects them from palmitate-induced apoptosis by regulating mitochondrial dynamics and biogenesis. MSI-1436 also increases glucose uptake and protects HPCs from palmitate-induced stress by reorganizing the cells' morphological architecture. Furthermore, our findings suggest that MSI-1436 enhances 2-NBDG uptake by increasing the expression of SIRT1, which is associated with liver insulin sensitivity. It also promotes mitochondrial dynamics by modulating mitochondria quantity and morphotype as well as increasing the expression of PINK1, MFN1, and MFN2. Our study provides evidence that MSI-1436 has a positive impact on equine hepatic progenitor cells, indicating its potential therapeutic value in treating EMS and insulin dysregulation.

Tracking the micro- and nanoplastics in the terrestrial-freshwater food webs. Bivalves as sentinel species.

Micro- (MPs) and nanoplastics (NPs) are currently ubiquitous in the ecosystems, and freshwater biota is still insufficiently studied to understand the global fate, transport paths, and consequences of their presence. Thus, in this study, we investigated the role of bivalves and a trophic transfer of MPs and NPs in an experimental food chain. The food chain consisted of terrestrial non-selective detritivore Dendrobaena (Eisenia) sp., freshwater benthic filter feeder Unio tumidus, and freshwater benthic detritivore-collectors Asellus aquaticus or Gammarus sp. Animals were exposed to different fluorescently labeled micro- and nanoplastics (PMMA 20 µm, nanoPS 15-18 nm, and 100 nm, PS 1 µm and 20 µm, PE from cosmetics) as well as to the faeces of animals exposed to plastics to assess their influence on the environmental transportation, availability to biota, and bioaccumulation of supplied particles. Damaged and intact fluorescent particles were observed in the faeces of terrestrial detritivores and in the droppings of aquatic filter feeders, respectively. They were also present in the guts of bivalves and of crustaceans which were fed with bivalve droppings. Bivalves (Unio tumidus, and additionally Unio pictorum, and Sphaerium corneum) produced droppings containing micro- and nanoparticles filtered from suspension and deposited them onto the tank bottom, making them available for broader feeding guilds of animals (e.g. collectors, like crustaceans). Finally, the natural ageing of PS and its morphological changes, leakage of the fluorescent labelling, and agglomeration of particles were demonstrated. That supports our hypothesis of the crucial role of the characterization of physical and chemical materials in adequately understanding the mechanisms of their interaction with biota. Microscopical methods (confocal, fluorescent, scanning electron) and Raman and FT-IR spectroscopy were used to track the particles' passage in a food web and monitor structural changes of the MPs' and NPs' surface.

The comparative study by Raman spectroscopy of the plastic tide in the three ports of the Mediterranean Sea.

This paper summarizes the field studies on marine microplastics (MPs) carried out in the autumn season in four various localisations within three ports chosen at the Mediterranean Sea near the French Riviera and the West Coast of Italy (within the Ligurian Sea). It considers the transport problem and the fate of the MPs introduced to the sea by analysing beach debris found on the shore after the stormy weather. Monitored ports included Saint-Tropez, Portoferraio and Porto Ercole, in which two different places were monitored. The aim is to approach the plastic tide phenomena by concentrating on a selected fraction of all MPs presented on the seashore. The final identification of debris was performed using Raman spectroscopy, providing a high-resolution signal. The PE, PP and PS contents were compared as the most frequent and representative polymers. Finally, we tackle the pending issue of the compound leakage from the MPs taking the environmentally aged particles from Portoferraio for further laboratory experiments and discuss an innovative approach with a low detection limit based on the electrochemical methods.

The impact of various carbon nanomaterials on the morphological, behavioural, and biochemical parameters of rainbow trout in the early life stages.

With the increasing production and the number of potential applications of carbon nanomaterials, mainly from the graphene family, their release into the natural environment, especially to aquatic ecosystems, is inevitable. The aim of the study was to determine the effects of various carbon nanomaterials (graphene nanoflakes (GNF), graphene oxide (GO), reduced graphene oxide (RGO) and silicon carbide nanofibers (NFSiC) in the concentration of 4 mg L-1 on the early life stages of the rainbow trout Oncorhynchus mykiss. The survival rates of O. mykiss were not affected after 36 days of exposure to studied materials, except for RGO, which caused significant mortality of both embryos and larvae compared to the control conditions. Larvae exposed to GO and NFSiC were characterized by a smaller standard body length at hatch, whereas at the end of the experiment, the growth of fish exposed to all materials was accelerated, especially in GO and RGO treatment, in which higher body weight and length were accompanied by lower volume of the yolk sac. Neither the markers of the oxidative damage nor the antioxidant enzymes activities were significantly affected in embryos, newly hatched larvae and larvae after 26-day exposure to studied carbon nanomaterials. Also, no neurotoxic effect expressed by the activity of the whole-body acetylcholinesterase was observed. Nevertheless, the significant increase in the velocity and the overall activity of larvae exposed to GNF (not investigated after exposure to other materials) must be highlighted. The most pronounced effect of RGO might be connected with its large particle size, sharp edges, and the presence of TiO2 nanoparticles. The results indicate for the first time that various carbon nanomaterials potentially released into aquatic ecosystems may have serious developmental implications for the early life stages of salmonid fish.