Amphibian peptides for skin protection and healing

BACKGROUND: Amphibians are currently suffering a dramatic decline worldwide, mainly due to chytridiomycosis, a skin infection caused by the pathogenic fungus Batrachochytrium dendrobatidis (Bd). An important natural defense of amphibian skin is the production of antimicrobial peptides (AMPs) by granular glands in the dermis. AMPs collected from several species of frogs successfully inhibit the growth of Bd in vitro. Besides their anti-microbial and anti-fungal activities, AMPs have been shown to exert other biological effects such as anti-viral, anti-tumor, anti-oxidant, immunomodulating and wound healing. AIM: We intended to test the efficacy of AMPs as cutaneous defenses in frog species either resistant or susceptible to Bd. METHODS: 3 frog species (Gastrotheca nebulanastes (GN), G. excubitor (GE) and Hypsiboas gladiator (HG), were collected in montane scrub, cloud forest and high elevation grassland habitats near Manu National Park in southeastern Peru. AMP secretion was stimulated by injection of norepinephrine into the dorsal lymph sacks. AMPs were then purified by chromatographic techniques. The human endothelial cell line HECV was treated with AMP concentrations ranging from 0.005 to 50 \ub5g/mL. Cell viability was verified by MTT test. Wound healing properties were analyzed by scratch wound assay. AMP inhibition strength against Bd growth was measured in vitro by incubating Bd zoospores with different concentrations of AMPs. RESULTS: Treatment with AMPs secreted from GN, GE and HG did not affect HECV cell viability at any concentration tested. No significant differences in cell migration rate were observed in HECV cells scratched and treated with GN and GE AMPs. Only HG peptides showed wound healing properties as well as strong Bd growth inhibiting ability. CONCLUSIONS: Stimulation of wound healing mechanisms and inhibition of Bd growth by skin AMPs might both contribute to HG resistance to chytridiomycosis. Understanding the role of skin defenses may lead to the development of novel Bd mitigation strategies. Possible applications of amphibian AMPs in skin medicine deserve attention and further studies. This work was funded by the European Commission (Tender ENV.B.3/SER/2016/0028, Mitigating a new infectious disease in salamanders to counteract the loss of European biodiversity) and by Parco Nazionale delle Cinque Terre

Immunomodulation by Different Types of N-Oxides in the Hemocytes of the Marine Bivalve Mytilus galloprovincialis

The potential toxicity of engineered nanoparticles (NPs) for humans and the environment represents an emerging issue. Since the aquatic environment represents the ultimate sink for NP deposition, the development of suitable assays is needed to evaluate the potential impact of NPs on aquatic biota. The immune system is a sensitive target for NPs, and conservation of innate immunity represents an useful basis for studying common biological responses to NPs. Suspension-feeding invertebrates, such as bivalves, are particularly at risk to NP exposure, since they have extremely developed systems for uptake of nano and microscale particles integral to intracellular digestion and cellular immunity. Evaluation of the effects of NPs on functional parameters of bivalve immunocytes, the hemocytes, may help understanding the major toxic mechanisms and modes of actions that could be relevant for different NP types in aquatic organisms.In this work, a battery of assays was applied to the hemocytes of the marine bivalve Mytilus galloprovincialis to compare the in vitro effects of different n-oxides (n-TiO2, n-SiO2, n-ZnO, n-CeO2) chosen on the basis of their commercial and environmental relevance. Physico-chemical characterization of both primary particles and NP suspensions in artificial sea water-ASW was performed. Hemocyte lysosomal and mitochondrial parameters, oxyradical and nitric oxide production, phagocytic activity, as well as NP uptake, were evaluated. The results show that different n-oxides rapidly elicited differential responses hemocytes in relation to their chemical properties, concentration, behavior in sea water, and interactions with subcellular compartments. These represent the most extensive data so far available on the effects of NPs in the cells of aquatic organisms. The results indicate that Mytilus hemocytes can be utilized as a suitable model for screening the potential effects of NPs in the cells of aquatic invertebrates, and may provide a basis for future experimental work for designing environmentally safer nanomaterials

3,5-Diiodo-L-thyronine modulates the expression of genes of lipid metabolism in a rat model of fatty liver.

Recent reports demonstrated that 3,5-diiodo-l-thyronine (T(2)) was able to prevent lipid accumulation in the liver of rats fed a high-fat diet (HFD). In this study, we investigated how the rat liver responds to HFD and T(2) treatment by assessing the transcription profiles of some genes involved in the pathways of lipid metabolism: oxidation, storage and secretion. The mRNA levels of the peroxisome proliferator-activated receptors (PPARα, PPARγ and PPARδ), and of their target enzymes acyl-CoA oxidase and stearoyl-CoA desaturase were evaluated by real-time RT-PCR. Moreover, the expression of the adipose triglyceride lipase involved in lipid mobilisation, of the main PAT proteins acting in lipid droplet (LD) turnover, and of apoprotein B (apo B), the major protein component of very low-density lipoproteins (VLDLs) were analysed. Overall, our data demonstrated that T(2) administration to HFD rats counteracts most of the hepatic transcriptional changes that occurred in response to the excess exogenous fat. In particular, our results suggest that T(2) may prevent the pathways leading to lipid storage in LDs, promote the processes of lipid mobilisation from LDs and secretion as VLDL, in addition to the stimulation of pathways of lipid oxidation. In conclusion, our findings might give an insight into the mechanisms underlying the anti-steatotic ability of T(2) and help to define the potential therapeutic role of T(2) for preventing or treating liver steatosis

Liquid and solid functional bio-based coatings

The development of new bio-based coating materials to be applied on cellulosic and plastic based substrates, with improved performances compared to currently available products and at the same time with improved sustainable end of life options, is a challenge of our times. Enabling cellulose or bioplastics with proper functional coatings, based on biopolymer and functional materials deriving from agro-food waste streams, will improve their performance, allowing them to effectively replace fossil products in the personal care, tableware and food packaging sectors. To achieve these challenging objectives some molecules can be used in wet or solid coating formulations, e.g., cutin as a hydrophobic water-and grease-repellent coating, polysaccharides such as chitosan-chitin as an antimicrobial coating, and proteins as a gas barrier. This review collects the available knowledge on functional coatings with a focus on the raw materials used and methods of dispersion/application. It considers, in addition, the correlation with the desired final properties of the applied coatings, thus discussing their potential

Immunotoxicity of polystyrene nanoplastics in different hemocyte subpopulations of Mytilus galloprovincialis

Plastic represents 60-80% of litter in the ocean. Degradation of plastic to small fragments leads to the formation of microplastics (MPs <5mm) and nanoplastics (NPs <1 mu m). One of the most widely used and representative plastics found in the ocean is polystyrene (PS). Among marine organisms, the immune system of bivalves is recognized as suitable to assess nanomaterial toxicity. Hemocyte subpopulations [R1 (large granular cells), R2 (small semi-granular cells) and R3 (small agranular or hyaline cells)] of Mytilus galloprovincialis are specialized in particular tasks and functions. The authors propose to examine the effects of different sizes (50 nm, 100 nm and 1 mu m) PS NPs on the different immune cells of mussels when they were exposed to (1 and 10mg.L-1) of PS NPs. The most noteworthy results found in this work are: (i) 1 mu m PS NPs provoked higher immunological responses with respect to 50 and 100nm PS NPs, possibly related to the higher stability in size and shape in hemolymph serum, (ii) the R1 subpopulation was the most affected with respect to R2 and R3 concerning immunological responses and (iii) an increase in the release of toxic radicals, apoptotic signals, tracking of lysosomes and a decrease in phagocytic activity was found in R1

Addressing nanomaterial immunosafety by evaluating innate immunity across living species

The interaction of a living organism with external foreign agents is a central issue for its survival and adaptation to the environment. Nanosafety should be considered within this perspective, and it should be examined that how different organisms interact with engineered nanomaterials (NM) by either mounting a defensive response or by physiologically adapting to them. Herein, the interaction of NM with one of the major biological systems deputed to recognition of and response to foreign challenges, i.e., the immune system, is specifically addressed. The main focus is innate immunity, the only type of immunity in plants, invertebrates, and lower vertebrates, and that coexists with adaptive immunity in higher vertebrates. Because of their presence in the majority of eukaryotic living organisms, innate immune responses can be viewed in a comparative context. In the majority of cases, the interaction of NM with living organisms results in innate immune reactions that eliminate the possible danger with mechanisms that do not lead to damage. While in some cases such interaction may lead to pathological consequences, in some other cases beneficial effects can be identified

Optimization of cognitive assessment in Parkinsonisms by applying artificial intelligence to a comprehensive screening test.

The assessment of cognitive deficits is pivotal for diagnosis and management in patients with parkinsonisms. Low levels of correspondence are observed between evaluations assessed with screening cognitive tests in comparison with those assessed with in-depth neuropsychological batteries. A new tool, we named CoMDA (Cognition in Movement Disorders Assessment), was composed by merging Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), and Frontal Assessment Battery (FAB). In total, 500 patients (400 with Parkinson's disease, 41 with vascular parkinsonism, 31 with progressive supranuclear palsy, and 28 with multiple system atrophy) underwent CoMDA (level 1-L1) and in-depth neuropsychological battery (level 2-L2). Machine learning was developed to classify the CoMDA score and obtain an accurate prediction of the cognitive profile along three different classes: normal cognition (NC), mild cognitive impairment (MCI), and impaired cognition (IC). The classification accuracy of CoMDA, assessed by ROC analysis, was compared with MMSE, MoCA, and FAB. The area under the curve (AUC) of CoMDA was significantly higher than that of MMSE, MoCA and FAB (p < 0.0001, p = 0.028 and p = 0.0007, respectively). Among 15 different algorithmic methods, the Quadratic Discriminant Analysis algorithm (CoMDA-ML) showed higher overall-metrics performance levels in predictive performance. Considering L2 as a 3-level continuous feature, CoMDA-ML produces accurate and generalizable classifications: micro-average ROC curve, AUC = 0.81; and AUC = 0.85 for NC, 0.67 for MCI, and 0.83 for IC. CoMDA and COMDA-ML are reliable and time-sparing tools, accurate in classifying cognitive profile in parkinsonisms.This study has been registered on ClinicalTrials.gov (NCT04858893)

Hyperbranched Quasi-1D TiO2 Nanostructure for Hybrid Organic-Inorganic Solar Cells

The performance of hybrid solar cells is strongly affected by the device morphology. In this work we demonstrate a Poly(3-hexylthiophene-2,5-diyl)/TiO2 hybrid solar cell where the TiO2 photoanode comprises an array of tree-like hyperbranched quasi-1D nanostructures self-assembled from the gas phase. This advanced architecture enables us to increase the power conversion efficiency to over 1%, doubling the efficiency with respect to state of the art devices employing standard mesoporous titania photoanodes. This improvement is attributed to several peculiar features of this array of nanostructures: high interfacial area; increased optical density thanks to the enhanced light scattering; and enhanced crystallization of Poly(3-hexylthiophene-2,5-diyl) inside the quasi-1D nanostructure

Massively Parallel RNA Sequencing Identifies a Complex Immune Gene Repertoire in the lophotrochozoan Mytilus edulis

The marine mussel Mytilus edulis and its closely related sister species are distributed world-wide and play an important role in coastal ecology and economy. The diversification in different species and their hybrids, broad ecological distribution, as well as the filter feeding mode of life has made this genus an attractive model to investigate physiological and molecular adaptations and responses to various biotic and abiotic environmental factors. In the present study we investigated the immune system of Mytilus, which may contribute to the ecological plasticity of this species. We generated a large Mytilus transcriptome database from different tissues of immune challenged and stress treated individuals from the Baltic Sea using 454 pyrosequencing. Phylogenetic comparison of orthologous groups of 23 species demonstrated the basal position of lophotrochozoans within protostomes. The investigation of immune related transcripts revealed a complex repertoire of innate recognition receptors and downstream pathway members including transcripts for 27 toll-like receptors and 524 C1q domain containing transcripts. NOD-like receptors on the other hand were absent. We also found evidence for sophisticated TNF, autophagy and apoptosis systems as well as for cytokines. Gill tissue and hemocytes showed highest expression of putative immune related contigs and are promising tissues for further functional studies. Our results partly contrast with findings of a less complex immune repertoire in ecdysozoan and other lophotrochozoan protostomes. We show that bivalves are interesting candidates to investigate the evolution of the immune system from basal metazoans to deuterostomes and protostomes and provide a basis for future molecular work directed to immune system functioning in Mytilus