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Graphene-based functional coatings for cotton textiles were realized through an easy dip-coating procedure. Cotton fabrics were coated with a reduced graphene oxide (rGO) layer and then protected with a very thin polyurethane (PU) layer that does not affect the flexibility and the hand of the pristine cotton. The application of the rGO coating induces electrical conductivity to the fabric and the application of the PU phase increases the durability of the coatings, that show very stable surface resistivity after 10 washing cycles performed at temperatures up to 40 °C. Furthermore, the rGO and rGO/PU coated fabrics show good comfort properties, increased thermal conductivity and breathability with respect to cotton. In particular, the realized coatings allow to confine the heat transfer in correspondence of a localized heating source, which is very interesting for thermal therapy applications. Finally, the rGO/PU coated fabrics present a piezoresistive behaviour characterized by very stable electrical response to applied stretching up to 50% deformation, high sensitivity especially at low deformations with gauge factor values up to 11.7 and fast response time down to 500 ms when stretched at 100 mm/min rate at 2.5% strain. Overall, the results demonstrate that rGO/PU coated fabrics are very promising wash-durable electrically conductive e-textiles with improved comfort, enhanced thermal conductivity for possible thermal therapy applications, and piezoresistive properties for sensing applications as human motion monitoring.

Microplastics have become a topic of considerable concern and intensive study over the past decade. They have been found everywhere in the oceans, including the deepest trenches and remotest parts of the Arctic. They are ingested by many animals and some are incorporated into tissues. There is considerable effort in studying what effects they have on marine life. It has become clear that when water samples are collected in ways that prevent most long thin particles from escaping through pores of a net, the most abundant type of microplastics found in water and sediments are microfibers (fibers with dimensions less than 5 mm). The major source of these pollutants is synthetic textiles, such as polyester or polyamides, which shed microfibers during their entire life cycle. Microfibers are released during textile manufacturing, everyday activities (e.g., washing, drying, wearing) and final disposal. The complexity of microfiber release mechanisms and of the factors involved make the identification and application of ways to reduce the inputs of microfibers very challenging. A comprehensive approach is strongly needed, taking into account solutions at a number of levels, such as re-engineering textiles to minimize shedding, applying washing machine filters, developing advanced wastewater treatment plants and improving the management of textile wastes. To harmonize and make mandatory the solutions identified, a variety of potential government policies and regulations is also needed.

The increasing plastic pollution of the world’s oceans represents a serious threat to marine ecosystems and has become a well-known topic garnering growing public attention. The global input of plastic waste into the oceans is estimated to be approximately 10 million tons per year and predicted to rise by one order of magnitude by 2025. More than 90% of the plastic that enters the oceans is thought to end up on the seafloor, and seafloor sediment samples show that plastics are concentrated in confined morphologies and sedimentary environments such as submarine canyons. These canyons are occasionally flushed by powerful gravity-driven sediment flows called turbidity currents, which transport vast volumes of sediment to the deep sea and deposit sediment in deep-sea fans. As such, turbidity currents may also transport plastics present in the canyon and bury plastics in deep-sea fans. These fans may therefore act as sinks for seafloor plastics. Here we present a comprehensive dataset showing the spatial distribution of microplastics in seafloor sediments from the Congo Canyon, offshore West Africa. Multicores taken from 16 locations along the canyon, sampled different sedimentary sub-environments including the canyon thalweg, canyon terraces, and distal lobe. Microplastics were extracted from the sediments by density separation and the polymer type, size, and shape of all individual microplastic particles were analysed using laser-direct infrared-spectroscopy (LDIR). Microplastic number concentrations in the sediments of the distal lobe are significantly higher than in the canyon, indicating that the Congo Canyon system is a highly efficient conduit for microplastic transport to the deep sea. Moreover, microplastic concentrations of >20,000 particles per kg of dry sediment were recorded in the lobe, which represent some of the highest ever recorded microplastic number concentrations in seafloor sediments. This shows that deep-sea fans can serve as hotspots and potential terminal sinks for seafloor microplastics.

Early life stages of marine fish populations may be strongly affected by environmental factors. Changes in the physical environment or the availability of food resources could lead to stress-related physiological responses affecting larval fitness, growth and survival. In the present study, we determined, for the first time, amino acid composition (AAC), lipid, and carbohydrate content, as well as alkaline phosphatase and peroxidase activities in larvae from the European anchovy Engraulis encrasicolus. Fishes were caught in two different spawning areas of the Strait of Sicily, characterized by different environmental conditions, including a coastal upwelling with a lower temperature (Adventure Bank; 20.22±0.38°C) and a thermohaline front with a higher temperature (Maltese Bank 23.10±0.25°C). The results showed that the two groups of larvae, in their early life, had similar nutritional status. However, compared with the samples from the Maltese Bank, the specimens collected in the Adventure Bank area exhibited higher alkaline phosphatase activity, lower concentrations of aspartate plus asparagine, threonine, and arginine but a higher concentration of leucine, highlighting different patterns of amino acid metabolism. Collectively, these results indicated that AAC analysis could represent an additional valid tool to evaluate the link between physiological responses and environmental conditions at early life stages.

Ovine papillomavirus (OaPV) comprises four genotypes; OaPV1, OaPV2, and OaPV4 are fibropapillomaviruses within the genus Delta-papillomavirus ( Delta-PV ), whereas OaPV3 is an epitheliotropic virus that belongs to the genus Dyokappa-papillomavirus ( Dyokappa-PV ). To date, all of them have been known to infect sheep only. OaPV1, OaPV2, and OaPV4 have been associated with ovine cutaneous and mucosal fibropapillomas, while OaPV3 is a key factor in the squamous cell carcinoma (SCC) pathway of the sheep skin. Peripheral blood mononuclear cell (PBMC) samples obtained from 128 cattle at public slaughterhouses were investigated using droplet digital polymerase chain reaction (ddPCR). ddPCR is a new-generation PCR technique that enables accurate and absolute quantification of target molecules with high sensitivity and specificity. All OaPVs were detected by identification and quantification of nucleic acids using specific fluorescent probes. Of 128 PBMC samples, 100 (~78%) showed OaPV infections. Further, 42, 35, and 23 PBMC samples showed single, double, and triple OaPV infections, respectively. OaPV1 was responsible for 22 single infections, OaPV2 caused 16 single infections, and OaPV3 and OaPV4 caused two single infections each. OaPV1 and OaPV2 were the most frequent ovine viruses in dual and triple infections. In many PBMC samples, both ovine Delta-PV and Dyokappa-PV were found to be transcriptionally active, as shown by the detection and quantification of E5 oncogene transcripts for OaPV1, L1 transcripts for OaPV2, E6 and E7 transcripts for OaPV3, and E6 for OaPV4. OaPVs were found in the blood samples from cattle that shared grasslands rich in bracken ferns known to contain immunosuppressant substances. Furthermore, OaPVs were also found in cattle from intensive livestock farming without any contact with sheep. Because OaPV DNA was detected in both grass hay and corn silage, it is conceivable that these feed may be the viral sources.

Microplastics released from textiles during the washing process represent the most prevalent type of microparticles found in different environmental compartments and ecosystems around the world. Release of microfibres during the washing process of synthetic textiles is due to the mechanical and chemical stresses that clothes undergo in washing machines. Several washing process parameters, conditions, formulations of laundering additives have been correlated to microfibre release and some of them have been identified to affect microfibre release during washing process, while no correlation has been evaluated between microfibre release and washing load. In the present study, microfibre release was evaluated as function of the washing load in a real washing process, indicating a progressive decrease of microfibre release with increasing washing load. The quantity of released microfibres increased by around 5 times by decreasing the washing load due to a synergistic effect between water-volume to fabric ratio and mechanical stress during washing. Moreover, the higher mechanical stress to which the fabric is subjected in the case of a low washing load, hinders the discrimination of the effect on the release of other washing parameters like the type of detergent and laundry additives used.

The washing process of synthetic fabrics represents the main source of primary microplastics in sea and oceans as large amounts of microfibers are released during washing. Recent researches have been focused on the development of effective approaches to mitigate the effects of microfiber release. Relevant approaches are aimed to design capturing devices and filters. In this work, the development, testing, and demonstration of the effectiveness of a new filtration system for washing machines are reported. This system is designed to retain microfibers avoiding their entrance into the wastewater system. Washing tests were performed using two different prototypes of the proposed filtration system, in order to optimize its design and efficiency. The obtained results demonstrate that, after its optimized, the proposed filtration system shows a capture efficiency of 64% toward microfibers released from commercial polyester t-shirts, comparable or even better than the efficiency shown by other commercial or prototype microfiber capturing systems.

Choroideremia (CHM) is a X-linked recessive chorioretinal dystrophy due to deficiency of the CHM gene product, i.e., Rab escort protein isoform 1 (REP1). To date, gene therapy for CHM has shown variable effectiveness, likely because the underlying pathogenic mechanisms as well as genotype-phenotype correlation are not yet fully known. Small nucleotide variants leading to premature termination codons (PTCs) are a major cause of CHM, but about 20% of patients has CHM gene deletions. To improve understanding of the disease mechanisms, we analyzed molecular features of seven deletions involving the CHM gene sequence. We mapped the deletion breakpoints by using polymerase chain reaction, sequencing and array comparative genomic hybridization, to identify rearrangement-promoting DNA sequences, we analyzed genomic architecture surrounding the breakpoint regions. Moreover, in some CHM patients with different mutation types, we measured transcript level of CHM and of CHML, encoding the REP2 isoform. Scattered along the whole CHM gene and in close proximity to the deletion breakpoints we found numerous repeat elements that generate a locus-specific rearrangement hot spot. Unexpectedly, patients with non-PTC variants had increased expression of the aberrant CHM mRNA, CHML expression was higher than normal in a patient lacking CHM and its putative regulatory sequences. This latest evidence suggests that mechanisms regulating CHM and CHML gene expression are worthy of further study, because their full knowledge could be also useful for developing effective therapies for this hitherto untreatable inherited retinal degeneration.

Persistent infection and tumourigenesis by papillomaviruses (PVs) require viral manipulation of various of cellular processes, including those involved in innate immune responses. Herein, we showed that bovine PV (BPV) E5 oncoprotein interacts with a tripartite motif-containing 25 (TRIM25) but not with Riplet in spontaneous BPV infection of urothelial cells of cattle. Statistically significant reduced protein levels of TRIM25, retinoic acid-inducible gene I (RIG-I), and melanoma differentiation-associated gene 5 (MDA5) were detected by Western blot analysis. Real-time quantitative PCR revealed marked transcriptional downregulation of RIG-I and MDA5 in E5-expressing cells compared with healthy urothelial cells. Mitochondrial antiviral signalling (MAVS) protein expression did not vary significantly between diseased and healthy cells. Co-immunoprecipitation studies showed that MAVS interacted with a protein network composed of Sec13, which is a positive regulator of MAVS-mediated RLR antiviral signalling, phosphorylated TANK binding kinase 1 (TBK1), and phosphorylated interferon regulatory factor 3 (IRF3). Immunoblotting revealed significantly low expression levels of Sec13 in BPV-infected cells. Low levels of Sec13 resulted in a weaker host antiviral immune response, as it attenuates MAVS-mediated IRF3 activation. Furthermore, western blot analysis revealed significantly reduced expression levels of pTBK1, which plays an essential role in the activation and phosphorylation of IRF3, a prerequisite for the latter to enter the nucleus to activate type 1 IFN genes. Our results suggested that the innate immune signalling pathway mediated by RIG-I-like receptors (RLRs) was impaired in cells infected with BPVs. Therefore, an effective immune response is not elicited against these viruses, which facilitates persistent viral infection.

Washing processes of synthetic clothes are considered the main source of microplastic pollution in the oceans, as during washing, microfibres are released and cannot be completely blocked by wastewater treatments plants, thus reaching the marine ecosystems. The development of mitigation approaches is strongly needed to prevent the impact of microplastics on marine environments. With the aim to protect the fabrics during the washing processes and then reduce the release of microfibres, in this work, new finishing treatments of polyamide fabrics were performed by an ElectroFluidoDynamic (EFD) method. This approach was used to obtain homogeneous coatings of biodegradable polymers, namely poly (lactic acid) and poly (butylene succinate-co-butylene adipate), on polyamide surface, without affecting the fabric hand and wettability. The treatments were very effective, able to significantly reduce the amount of microfibres released during washing tests, proving to be a promising application to mitigate micro plastic pollution. (C) 2019 Elsevier Ltd. All rights reserved.

E5 protein, the major oncoprotein of the bovine Deltapapillomavirus genus, has been detected in 17 of the 19 urothelial cancers by molecular and morphological procedures. In 10 urothelial cancers, the oxygen sensitive subunit HIF-1α, which is upregulated by hypoxia, was overexpressed. Mitophagy, the selective autophagic removal of dysfunctional mitochondria, was upregulated in hypoxic neoplastic cells infected by BPVs which was mediated by FUNDC1, a mitochondrial outer-membrane protein. The FUNDC1 receptor was amplified by PCR, and amplicon sequencing showed a 100% homology with bovine FUNDC1 sequences deposited in GenBank (accession number: NM_001104982). Both transcripts and protein levels of FUNDC1 were significantly decreased in hypoxic neoplastic cells relative to healthy, non-neoplastic cells. FUNDC1 interacted with the LC3 protein, a marker of autophagosome (mitophagosome) membrane, the Hsc70/Hsp70 chaperone, and Bag3 co-chaperone. Bag3 may play a role in mitophagosome formation together with the Synpo2 protein, and may be involved in the degradation of Hsc70/Hsp70-bound CHIP-ubiquitinated cargoes, in association with its chaperone. Ultrastructural findings revealed the presence of mitochondria exhibiting severe fragmentation and loss of cristae, as well as numerous mitochondria-containing autophagosomes. Total and phosphorylated GTPase dynamin-related protein 1 (DRP1), which plays a crucial role in mitochondrial fission, a pre-requisite for mitophagy, was overexpressed at the mitochondrial level. Total and phosphorylated mitochondrial fission factor (Mff), mitochondrial fission protein 1 (Fis1), mitochondrial dynamics 51 (MiD51), and MiD49, which are DRP1 receptors responsible and/or co-responsible for its mitochondrial recruitment were overexpressed.

E5 protein, the major oncoprotein of bovine Deltapapillomavirus (BPV), was found to be expressed in 18 of 21 examined urothelial cancers of cattle. E5 oncoprotein was found to interact with p62 which was degraded through the autophagosome-lysosome pathway as well as LC3-II and appeared to be involved in the phosphorylation of the α-subunit of eukaryotic initiation factor 2 (eIF2α). Autophagy was morphologically documented by transmission electron microscope (TEM) through the detection of double-membrane autophagosomes and autolysosomes. Overexpression of Bag3 known to mediate selective autophagy was also demonstrated. Furthermore, Bag3 and BPV E5 oncoprotein were seen to co-localize with dynein and 14-3-3γ, which suggested that Bag3 could be involved in inducing the retrograde transport of BPV E5 along microtubules to aggresomes, perinuclear sites with high autophagic flux. Electron dense perinuclear structures consistent with aggresomes were also documented by TEM in urothelial cancer cells. Finally, Bag3 was found to also interact with synaptopodin 2 (Synpo2), which would seem to contribute to cargo degradation as it has been shown to facilitate autophagosome formation. This study provides mechanistic insights into the potential role(s) of autophagy in BPV disease, which can help to develop future treatment and control measures for BPV infection. Activation of autophagy correlates positively with BPV infection and may play a role in biological behavior of bladder cancer as urothelial carcinomas of cattle are known to be characterized by a relatively low rate of metastasis.

Hypothesis In the last years, several cost-effective technologies have been investigated to functionalize textile substrates for large scale applications and industrial production. However, several limitations of currently used technologies still restrict the capability to form functional coatings finely controlling the textile surface properties and topographic structure of the coatings at sub-micrometric scale. Experiments Herein, we introduced a new non-conventional electrofluidodynamic technology – based on the use of electrostatic forces to polymer/composite solutions – for the application onto textile fabrics of functional coatings. With respect to particle/fibrous coatings usually applied through conventional electrospraying/electrospinning processes, the proposed approach is able to realize homogeneous and continuous coatings by a one-step process, imparting tailored functionalities to the textiles surfaces through the use of customized experimental setups. Findings We proved that this process can be successfully used to realize functional coatings based on a bioderived polymer, namely polylactic acid (PLA), on commercial woven polyamide (PA) fabrics. In addition, due to the usage of graphene derivatives or photochromic dyes in combination with PLA, the applied coatings are able to confer peculiar functionalities (i.e., electrical conductivity, photochromic properties, etc.) to polyamide fabrics, as proved by SEM, conductivity and UV irradiation measurements, for innovative applications in smart textiles, e-health and wearable electronics.

Vertical transmission of bovine papillomavirus (BPV) infection was investigated on livers and kidneys of four foetuses from cows suffering from BPV-2-associated urothelial cancers of the urinary bladder. PCR analysis revealed the presence of BPV-2 E5 DNA in the livers and kidneys of two foetuses. Amplified DNA fragments, composed of 502 bp, showed a 100% homology with BPV-2 sequences (GenBank accession number: M20219.1). BPV-2 was found to be transcriptionally active. Indeed, reverse transcriptase (RT)-PCR showed BPV-2 E5 transcripts. Sequencing of amplified cDNA, composed of 154 bp, showed a 100% identity with BPV-2 E5 sequences (GenBank accession number: M20219.1). Western blot analysis revealed the presence of dimers of E5 oncoprotein. Furthermore, a statistically significant increase of the phosphorylated (activated) form of the platelet-derived growth factor ß receptor (PDGFßR) was also detected in the fetal tissues. PDGFßR is believed to form the most important interaction with the E5 oncoprotein, thus regulating biological activity of virus protein. The strong concordance between virus found in fetal organs with virus detected in infected mothers provides evidence that BPV-2 can spread through blood and vertical infection occurs via transplacental transmission. Finally, molecular findings of this study raise unsolved questions about the potential role of BPVs in reproductive disorders. The presence of E5 oncoprotein, as in adult organs, may also activate the constitutive receptor PDGFßR in foetal organs, which plays a pivotal role in angiogenesis and embryonic development. Therefore, abnormal phosphorylation of PDGFßR may be involved in vascular and organogenesis abnormalities other than cancer.

Highly pathogenic bovine Delta papillomaviruses (δPVs) were detected and quantified for the first time using digital droplet polymerase chain reaction (ddPCR) by liquid biopsy in 103 clinically healthy sheep. Overall, ddPCR detected bovine δPVs in 68 blood samples (66%). Bovine papillomavirus (BPV) infection by a single genotype was revealed in 59% of the blood samples, and BPV coinfection by double, triple or quadruple genotypes was observed in 41% of liquid biopsies. The BPV-2 genotype was most frequently seen in sheep, whereas BPV-1 was the least common. Furthermore, ddPCR was very useful for detection and quantification; the BPV-14 genotype was observed for the first time in ovine species, displaying the highest prevalence in some geographical areas (Apulia). In 42 of the positive samples (61.8%), a single BPV infection was observed, 26 of which were caused by BPV-2 (61.9%) and 7 by BPV-13 (16.7%). BPV-14 was responsible for 7 single infections (16.7%) and BPV-1 for 2 single infections (4.7%). Multiple BPV coinfections were observed in the remaining 26 positive samples (38.2%), with dual BPV-2/BPV-13 infection being the most prevalent (84.6%). BPV infection by triple and quadruple genotypes was also observed in 11.5% and 3.8% of cases, respectively. The present study showed that ddPCR, a biotechnological refinement of conventional PCR, is by far the most sensitive and accurate assay for BPV detection. Therefore, ddPCR displayed diagnostic and epidemiological value resulting in the identification of otherwise undetectable BPV genotypes as well as their geographical distributions and suggesting that animal husbandry practices contribute to cross-species transmission of BPVs..

Ovine papillomaviruses (OaPVs) were detected and quantified, for the first time, using droplet digital polymerase chain reaction (ddPCR) and real-time quantitative PCR (qPCR) via blood samples of 165 clinically healthy sheep. OaPV DNA was detected in 126 blood samples (~76.4%). DdPCR detected OaPV DNA in 124 samples; in only two additional samples positive for real-time qPCR, ddPCR failed to detect the presence of any OaPVs. In 70 of the positive samples (~55.6%), a single OaPV infection was observed, 12 of which were caused by OaPV1 (~17.1%) and 14 by OaPV2 (20%). OaPV3 was responsible for 19 single infections (~27.1%), and OaPV4 for 25 single infections (~35.7%). Multiple OaPV coinfections were observed in 56 (~44.4%) positive samples. OaPV coinfections caused by two genotypes were observed in 31 positive samples (~55.4%), with dual OaPV3/OaPV4 infection being the most prevalent as seen in 11 blood samples. In addition, five OaPV1/OaPV4, four OaPV1/OaPV2, four OaPV2/OaPV3, four OaPV1/OaPV3, and three OaPV2/OaPV4 dual coinfections were also detected. OaPV coinfections by triple and quadruple genotypes were detected in 24 (~42.8%) and only one (~1.8%) of coinfected blood samples, respectively. Multiple infections caused by OaPV1/OaPV3/OaPV4 genotypes were the most prevalent, as observed in 12 (50%) blood samples harboring triple OaPV infections. This study showed that ddPCR is the most sensitive and accurate assay for OaPV detection and quantification thus outperforming real-time qPCR in terms of sensitivity and specificity. Therefore, ddPCR may represent the molecular diagnostic tool of choice, ultimately providing useful insights into OaPV molecular epidemiology and field surveillance.

Sandy beaches represent environmental compartments particularly vulnerable to litter pollution, and they reflect the magnitude of pollution of adjacent compartments: water and coastal areas. The substitution of conventional polymers by biodegradable materials is generally considered as an alternative for reducing environmental accumulation of plastic debris. The present study is aimed to investigate the degradation of poly(lactic acid), poly (?-caprolactone), poly(butylenesuccinate adipate) and poly(3-hydroxybutyrate) buried in sand for 267 days, simulating them as beach litter. The analysed polyesters showed different degradation mechanisms and kinetics. PLA is mainly subjected to weathering by physical aging; after an initial faster degradation of the amorphous phase, PCL showed a decrease of its degradation rate; similarly to PCL, the degradation of PBSA started from the amorphous phase; PHB is clearly subjected to biological degradation. The degradation trend of the investigated materials in sand decreased in the order PHB > PBSA > PCL > PLA. PLA, PCL and PBSA did not undergo complete degradation in sand during the testing time.

Over the last few decades, a large number of pH sensitive materials with new compositions and structures have been proposed. Solid state sensors based on organic, inorganic and composite materials are actively investigated, with an increasing interest in the performance offered by nano-scale materials. Our review provides a thorough, up-to-date knowledge of a wide range of pH measurement methods and related sensing materials, firstly by introducing well established materials and methods for pH sensing and then, by covering recent developments in inorganic, organic and nano-engineered devices. The main sensor parameters, including sensitivity, stability, response time and testing conditions are reported. Given the importance of pH sensing in environmental applications, in particular seawater monitoring, sensors tested in seawater are highlighted and discussed.

In the present study, the highly pathogenic bovine deltapapillomavirus (δPV) was investigated by liquid biopsy in blood samples of 168 clinically normal goats using both droplet digital PCR (ddPCR) and quantitative real-time PCR (qPCR). Overall, ddPCR discovered BPV E5 DNA in ~ 61.3% of the blood samples examined, while real-time qPCR revealed the virus in ~ 10.7% of the same samples. Moreover, ddPCR showed BPV E5 DNA in ~ 78.8% of blood samples from goats that were in close contact with cattle and in 20% of blood samples from goats living in closed pens without any contact with cattle. In addition, ddPCR unmasked a single BPV genotype in ~ 59.2% and multiple genotypes in ~ 40.8% of goats harbouring BPV DNA, while real-time qPCR detected single genotypes in ~ 17% and multiple genotypes in ~ 1%. Of the BPV co-infections detected by ddPCR, 28 (~67%) involved two genotypes, eight (~19%) three genotypes and six (~14%) four genotypes. In contrast, real-time qPCR revealed BPV co-infection by two genotypes in only one sample and failed to detect co-infection by three or four genotypes. BPV2 and BPV13 were the most prevalent viruses responsible for single and multiple co-infections, respectively. The present study showed that ddPCR is promising method for circulating bovine papillomavirus DNA detection and quantification and suggested that animal husbandry practices contribute to cross-species transmission of BPVs.

The assessment of the distribution and composition of microlitter in the sea is a great challenge. Biological indicators can be an irreplaceable tool since they measure microlitter levels in their environments in a way that is virtually impossible to replicate by direct physical measurements. Furthermore, trends can provide policymakers with statistically robust analysis. We looked into the capacity of multiple fish species to describe the distribution and composition of microlitter vertically across different compartments of the water column. A total of 502 individuals from six selected species (Scomber scombrus, Oblada melanura, Spicara smaris, Boops boops, Merluccius merluccius and Mullus barbatus) were collected on the western side of Sardinia island and allocated to three compartments: surface, mid-water and bottom. The species of the surface exhibited a higher frequency of occurrence (41.89%) of microlitter ingestion, compared to those of the mid-water and bottom (19.60%; 22.58%). A significant difference in the average number of ingested microlitter was found between the surface and the bottom compartment. All the microlitter fragments found were analysed through Fourier Transform Infrared Spectroscopy (FTIR). The comparison of the expected buoyancies of the polymers identified puth faith in the allocation of the species to the respective compartments. Therefore, considering the Marine Strategy Framework Directive objective, this approach could be useful in assessing microlitter distribution and composition vertically across the water column.

In this study, the digital droplet polymerase chain reaction (ddPCR) was used to quantify circulating bovine papillomavirus (BPV; genus: Deltapapillomavirus) DNA levels in blood samples from 25 clinically normal cows and 15 cows with chronic enzootic haematuria due to papillomavirus-associated bladder tumours. ddPCR detected BPV DNA in 95% of all the samples (i.e. in 24 of the clinically normal cows and 14 of the diseased animals), whereas quantitative real-time PCR (qPCR) detected it in only 57.5% of the same blood samples, with percentage differences between ddPCR and qPCR being statistically significant (p-value ≤ .05), according to chi-squared test. Furthermore, ddPCR detected BPV infections by a single genotype and by multiple genotypes in 37% and 63% of the cows, whereas qPCR detected these in 16% and 16%. Of the two assays, ddPCR was the more sensitive and accurate clinical diagnostic tool, allowing the detection of otherwise undetectable BPV genotypes, and consequently, a higher number of BPV co-infections. qPCR failed to detect many BPV co-infections by multiple genotypes. Therefore, ddPCR may be an essential tool for improving diagnostic procedures, allowing the identification of the genotypic distribution of BPV and a better understanding about the territorial divergence, if any, of the BPV prevalence in different areas. No significant differences in the blood viral load estimations were observed between the two animal groups, suggesting that the bloodstream could be a site of primary infection. Finally, as BPV DNA was detected in cows affected by non-invasive urothelial tumours, including papilloma and papillary urothelial neoplasms of low malignant potential, the circulating BPVs appeared to be independent of the status of urothelial neoplasms. Therefore, unlike in humans, circulating BPVs cannot be an actual prognostic marker of urothelial tumours in cows.

The washing of synthetic fabrics has been identified recently as one of the major contributors to the global release of primary microplastics to the oceans. The mechanical and chemical stresses of a washing process can cause the detachment of “microfibers” from the yarns composing a fabric, which cannot be completely removed by wastewater treatment plants (WWTPs). Also, WWTPs should have an important role as barriers for the entrance of microplastics into aquatic environments, particularly for microfibers from the washing of synthetic clothes. The Canadian company Environmental Enhancements is selling the Lint LUV-R Washing Machine Discharge Filter as a device capable of screening out synthetic microplastic particulates, but no studies on its actual efficiencies are available. In conclusion, an effective prevention of microplastic release from the washing processes of synthetic clothes can be achieved only by applying mitigation actions at different stages including textile design, finishing treatments, washing method, and the treatment at WWTPs.

This study aimed to provide mechanistic insights into mitophagy pathway associated with papillomavirus infection in urothelial cells of cattle. The elimination of mitochondria via autophagy, termed mitophagy, is an evolutionarily conserved mechanism for mitochondrial quality control and homeostasis. PINK1/parkin-mediated mitophagy, a ubiquitin-dependent selective autophagy of dysfunctional mitochondria, has been described here, for the first time, in urothelial cells from 25 bladder cancers in cattle infected by bovine papillomavirus (BPV). The expression of BPV-2 and BPV-13 E5 oncoprotein was detected by RT-PCR. Abnormal mitochondria delimited by expanding phagophores, were peculiar ultrastructural features of neoplastic urothelial cells. High levels of mitochondrial phosphorylated PINK1/parkin were observed in neoplastic urothelial cells infected by BPVs. Phosphoparkin interacted with mitofusin 2 (Mfn2) and ubiquitin (Ub), which confirmed that Mfn2 is a parkin receptor at the mitochondrial level, where parkin interacted also with Ub. Furthermore, parkin established a complex that was comprised of optineurin, p62, LC3, laforin, and embryonic stem cell-expressed Ras (ERAS), that interacted with BPV E5 oncoprotein, and Bag3, which, in turn, regulated the formation of a complex composed of Hpc70/Hsp70, CHIP, an HSC70-interacting E3 ubiquitin ligase. It is conceivable that ERAS is involved in mitophagosome maturation via phosphatidylinositol 3-kinase (PI3K) pathway. Bag3, in association with Hsc70/Hsp70, may contribute to the transport and degradation of CHIP-ubiquitinated cargo as this complex recognises ubiquitinated cargos and transports them to aggresomes to be degraded. Furthermore, Bag3 may be involved in mitophagosome formation as it interacted with synaptopodin 2, which is known to play a role in mitophagosome biogenesis.

This is the first study showing an in vivo microautophagy upregulation by Leishmania infantum in dogs. Both Leishmania amastigotes and promastigotes were detected in the cytoplasm of many professional and nonprofessional phagocytic cells of popliteal lymph node of three dogs suffering from chronic cutaneous leishmaniasis. Ultrastructurally, parasites appeared to be wrapped by lysosomes and/or multivesicular bodies. Neither phagophores nor double-membraned vacuoles consistent with autophagosomes were observed. Transcription factor EB (TFEB), a key factor involved in lysosome biogenesis, showed a statistically significant increase in the total component when examined by western blot in samples from leishmaniotic dogs compared with samples from healthy dogs. Instead, phosphorylated TFEB showed unmodified expression levels both in leishmaniotic and healthy dogs. Furthermore, Hsc70 and endosomal sorting complex required for transport (ESCRT)-I, which are known to play a role in microautophagy, showed no variation in expression levels both in diseased and healthy animals. Vps4A/B, an evolutionary conserved ATPase responsible for ESCRT-I complex disassembly and MVB maturation, was statistically significantly overexpressed in lymph nodal samples from leishmaniotic dogs. Bag3 was downregulated in diseased dogs whereas CHIP, p62, and LC3-II did not show any variation in expression levels. The altered expression profile of Bag3 suggested an altered interaction of Bag3 with Hsc70 and CHIP, which usually form a molecular complex involved in autophagosome-lysosome pathways. Ultrastructural and molecular findings suggested that the microautophagy pathway is upregulated in lymph nodes of dogs suffering from a chronic natural infection by Leishmania infantum.