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 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.

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.

Reversible rearrangement of the cellular cytoskeleton: A key to the broad-spectrum antiviral activity of novel amphiphilic polymers.

The battle against emerging viral infections has been uneven, as there is currently no broad-spectrum drug available to contain the spread of novel pathogens throughout the population. Consequently, the pandemic outbreak that occurred in early 2020 laid bare the almost empty state of the pandemic box. Therefore, the development of novel treatments with broad specificity has become a paramount concern in this post-pandemic era. Here, we propose copolymers of poly (sodium 2-(acrylamido)-2-methyl-1-propanesulfonate) (PAMPS) and poly (sodium 11-(acrylamido)undecanoate (AaU), both block (PAMPS75-b-PAaUn) and random (P(AMPSm-co-AaUn)) that show efficacy against a broad range of alpha and betacoronaviruses. Owing to their intricate architecture, these polymers exhibit a highly distinctive mode of action, modulating nano-mechanical properties of cells and thereby influencing viral replication. Through the employment of confocal and atomic force microscopy techniques, we discerned perturbations in actin and vimentin filaments, which correlated with modification of cellular elasticity and reduction of glycocalyx layer. Intriguingly, this process was reversible upon polymer removal from the cells. To ascertain the applicability of our findings, we assessed the efficacy and underlying mechanism of the inhibitors using fully differentiated human airway epithelial cultures, wherein near-complete abrogation of viral replication was documented. Given their mode of action, these polymers can be classified as biologically active nanomaterials that exploit a highly conserved molecular target-cellular plasticity-proffering the potential for truly broad-spectrum activity while concurrently for drug resistance development is minimal.

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 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.

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.

Low-volume label-free SARS-CoV-2 detection with the microcavity-based optical fiber sensor.

Accurate and fast detection of viruses is crucial for controlling outbreaks of many diseases; therefore, to date, numerous sensing systems for their detection have been studied. On top of the performance of these sensing systems, the availability of biorecognition elements specific to especially the new etiological agents is an additional fundamental challenge. Therefore, besides high sensitivity and selectivity, such advantages as the size of the sensor and possibly low volume of analyzed samples are also important, especially at the stage of evaluating the receptor-target interactions in the case of new etiological agents when typically, only tiny amounts of the receptor are available for testing. This work introduces a real-time, highly miniaturized sensing solution based on microcavity in-line Mach-Zehnder interferometer (µIMZI) induced in optical fiber for SARS-CoV-2 virus-like particles detection. The assay is designed to detect conserved regions of the SARS-CoV-2 viral particles in a sample with a volume as small as hundreds of picoliters, reaching the detection limit at the single ng per mL level.

The Raman spectroscopy and SEM/EDS investigation of the primary sources of microplastics from cosmetics available in Poland.

The increasing amount of marine microplastics and nanoplastics is due to primary (P-MPs) and secondary sources (S-MPs). Among small P-MPs and fibres from textiles, nurdles or industrial dust, the microbeads from cosmetics (peelings, scrubs) are dominant. The polyethylene and polypropylene debris, already <5 mm, enlarge Plastisphere and have a complex influence on environmental processes. Fortunately, the situation has changed with the limitation of synthetic materials in scrubs in recent years. This paper summarizes the six years of studies (2015-2021) on microplastics in cosmetics available on the Polish market. It focuses on those still available and presents the potential of Raman spectroscopy for the efficient qualitative and quantitative characterization of those materials. Finally, surface morphology's vital role in microplastics' general behaviour is underlined. The SEM/EDX and numerical modelling enables a more detailed description of particles.

SARS-CoV-2 infects an in vitro model of the human developing pancreas through endocytosis.

Recent studies showed that SARS-CoV-2 can infect adult human pancreas and trigger pancreatic damage. Here, using human fetal pancreas samples and 3D differentiation of human pluripotent cells into pancreatic endocrine cells, we determined that SARS-CoV-2 receptors ACE2, TMPRSS2, and NRP1 are expressed in precursors of insulin-producing pancreatic ß-cells, rendering them permissive to SARS-CoV-2 infection. We also show that SARS-CoV-2 enters and undergoes efficient replication in human multipotent pancreatic and endocrine progenitors in vitro. Moreover, we investigated mechanisms by which SARS-CoV-2 enters pancreatic cells, and found that ACE2 mediates the entry, while NRP1 and TMPRSS2 do not. Surprisingly, we found that in pancreatic progenitors, SARS-CoV-2 enters cells via cathepsin-dependent endocytosis, which is a different route than in respiratory tract. Therefore, pancreatic spheroids might serve as a model to study candidate drugs for endocytosis-mediated viral entry inhibition and to investigate whether SARS-CoV-2 infection may affect pancreas development, possibly causing lifelong health consequences.

Human Intramuscular Hyperimmune Gamma Globulin (hIHGG) Anti-SARS-CoV-2-Characteristics of Intermediates and Final Product.

This study aims to characterize the intermediates, and the final product (FP) obtained during the production of human intramuscular hyperimmune gamma globulin anti-SARS-CoV-2 (hIHGG anti-SARS-CoV-2) and to determine its stability. Material and methods: hIHGG anti-SARS-CoV-2 was fractionated from 270 convalescent plasma donations with the Cohn method. Prior to fractionation, the plasma was inactivated (Theraflex MB Plasma). Samples were defined using enzyme immunoassays (EIA) for anti-S1, anti-RBD S1, and anti-N antibodies, and neutralization assays with SARS-CoV-2 (VN) and pseudoviruses (PVN, decorated with SARS-CoV-2 S protein). Results were expressed as a titer (EIA) or 50% of the neutralization titer (IC50) estimated in a four-parameter nonlinear regression model. Results: Concentration of anti-S1 antibodies in plasma was similar before and after inactivation. Following fractionation, the anti-S1, anti-RBD, and anti-N (total tests) titers in FP were concentrated approximately 15-fold from 1:4 to 1:63 (1800 BAU/mL), 7-fold from 1:111 to 1:802 and from 1:13 to 1:88, respectively. During production, the IgA (anti-S1) antibody titer was reduced to an undetectable level and the IgM (anti-RBD) titer from 1:115 to 1:24. The neutralizing antibodies (nAb) titer increased in both VN (from 1:40 to 1:160) and PVN (IC50 from 63 to 313). The concentration of specific IgG in the FP did not change significantly for 14 months. Conclusions: The hIHGG anti-SARS-CoV-2 was stable, with concentration up to approximately 15-fold nAb compared to the source plasma pool.

The first evidence of the Indian meal moth (Plodia interpunctella) interaction with the silicone moulds.

The Indian meal moth (Plodia interpunctella, Lepidoptera) is a common insect species and well-known, widespread kitchen pest. This paper reports the results of their interaction with silicone baking moulds and the evidence for synthetic material decomposition. The obtained fibres exhibited a high level of purification and were characterized by Raman spectroscopy. The reported interaction should be further studied from the biological perspective, as it can be crucial for the remediation of silicones or other synthetic polymers in the environment.

Plant-Oil-Based Fibre Composites for Boat Hulls.

Plant-oil-based fibre composites for boat hulls are an interesting and growing group of materials. Although many problems are to be tackled at different stages of their preparation, the green composites are already successfully commercialised. Within this paper, the most important chemical and physical characteristics of both natural fibres and sustainable resins are provided in the form of a comprehensive review. Finally, the complex issue of interactions filler-matrix is considered.

Plastic in digestive tracts and gills of cod and herring from the Baltic Sea.

Plastic litter is widespread on our planet and is recognized as a contaminant of high concern. Plastic ingestion and retention in gills were studied in two key Baltic fish species: herring (Clupea harengus L.) and cod (Gadus morhua L.). In total, 183 fish from the southern Baltic Sea were analysed. Plastic litter was found in digestive tracts of 12.7 and 14.8% of herrings and cods, respectively. In addition, gills were shown to constitute an important transfer route of plastic to cod (9.9% of cods) but not to herring thus likely reflecting species-specific differences in fish lifestyles and/or water filtering capacity. No more than one plastic item per individual was found except for three fish with two items in their stomachs. Dominant microplastics (<5 mm) (MPs) (88.6% of all items) composed of irregular fragments, fibres and foils (61.3%, 25.8% and 12.9% of all MPs, respectively) were followed by meso- (5-25 mm) and macro-plastics (>25 mm) (5.7% each of all items). Plastic ingestion and retention in gills did not affect fish body condition assessed by Fulton's K index, although herrings that ingested plastic particles beyond the microplastic size range (i.e. >5 mm) were characterized by the lowest condition indices. All plastic items recovered from fish exhibited clear signs of weathering and surface extended due to their roughness and fragmentation which may enhance both sorption properties for chemical compounds and/or their leaching. Nevertheless, chemical contaminant levels in herring muscles were not different than in a plastic-free group. The vast majority of fibres found in fish was identified as air-borne or procedural contamination and were excluded from further analyses. In addition, the vessel's paint dust found in fish stressed the need for strict quality assurance also during fish sampling.

Acriflavine, a clinically approved drug, inhibits SARS-CoV-2 and other betacoronaviruses.

The COVID-19 pandemic caused by SARS-CoV-2 has been socially and economically devastating. Despite an unprecedented research effort and available vaccines, effective therapeutics are still missing to limit severe disease and mortality. Using high-throughput screening, we identify acriflavine (ACF) as a potent papain-like protease (PLpro) inhibitor. NMR titrations and a co-crystal structure confirm that acriflavine blocks the PLpro catalytic pocket in an unexpected binding mode. We show that the drug inhibits viral replication at nanomolar concentration in cellular models, in vivo in mice and ex vivo in human airway epithelia, with broad range activity against SARS-CoV-2 and other betacoronaviruses. Considering that acriflavine is an inexpensive drug approved in some countries, it may be immediately tested in clinical trials and play an important role during the current pandemic and future outbreaks.

Functional Severe Acute Respiratory Syndrome Coronavirus 2 Virus-Like Particles From Insect Cells.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains a major epidemic threat since the beginning of 2020. Efforts to combat the virus and the associated coronavirus disease 2019 (COVID-19) disease are being undertaken worldwide. To facilitate the research on the virus itself, a number of surrogate systems have been developed. Here, we report the efficient production of SARS-CoV-2 virus-like particles (VLPs) in insect cells. Contrary to widely used pseudovirus particles, where only one coronaviral protein is displayed within a heterologous scaffold, developed VLPs are structurally similar to the native virus and allow for more throughput studies on the biology of the infection. On the other hand, being devoid of the viral genome, VLPs are unable to replicate and thus safe to work with. Importantly, this is the first report showing that SARS-CoV-2 VLPs can be efficiently produced in insect cells and purified using scalable affinity chromatography.

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.

SARS-CoV-2 inhibition using a mucoadhesive, amphiphilic chitosan that may serve as an anti-viral nasal spray.

There are currently no cures for coronavirus infections, making the prevention of infections the only course open at the present time. The COVID-19 pandemic has been difficult to prevent, as the infection is spread by respiratory droplets and thus effective, scalable and safe preventive interventions are urgently needed. We hypothesise that preventing viral entry into mammalian nasal epithelial cells may be one way to limit the spread of COVID-19. Here we show that N-palmitoyl-N-monomethyl-N,N-dimethyl-N,N,N-trimethyl-6-O-glycolchitosan (GCPQ), a positively charged polymer that has been through an extensive Good Laboratory Practice toxicology screen, is able to reduce the infectivity of SARS-COV-2 in A549ACE2+ and Vero E6 cells with a log removal value of - 3 to - 4 at a concentration of 10-100 µg/ mL (p < 0.05 compared to untreated controls) and to limit infectivity in human airway epithelial cells at a concentration of 500 µg/ mL (p < 0.05 compared to untreated controls). In vivo studies using transgenic mice expressing the ACE-2 receptor, dosed nasally with SARS-COV-2 (426,000 TCID50/mL) showed a trend for nasal GCPQ (20 mg/kg) to inhibit viral load in the respiratory tract and brain, although the study was not powered to detect statistical significance. GCPQ's electrostatic binding to the virus, preventing viral entry into the host cells, is the most likely mechanism of viral inhibition. Radiolabelled GCPQ studies in mice show that at a dose of 10 mg/kg, GCPQ has a long residence time in mouse nares, with 13.1% of the injected dose identified from SPECT/CT in the nares, 24 h after nasal dosing. With a no observed adverse effect level of 18 mg/kg in rats, following a 28-day repeat dose study, clinical testing of this polymer, as a COVID-19 prophylactic is warranted.

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.