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The southern branch of the Elizabeth River near Portsmouth, Virginia, USA, is one of the most creosote-polluted subestuaries in North America and the former location of the Atlantic Wood US Environmental Protection Agency Superfund site. We previously demonstrated that adult Atlantic Wood killifish collected in situ had severe hepatic lesions, including hepatoblastoma and hepatocellular carcinoma, as well as suppressed circulating antibody responses compared to a historical reference site. Moreover, several innate immune functions were higher in Atlantic Wood fish, including elevated expression of hepatic cyclooxygenase-2 (COX-2), suggesting a proinflammatory environment. To further examine the potential of Atlantic Wood contaminants to modulate innate immune function(s), the present study used RAW264.7 mouse macrophages as an in vitro model to develop new approach methodologies for rapid screening. Lipopolysaccharide (LPS)-stimulated nitric oxide secretion by macrophages is a rapid, sensitive, and predictive in vitro system for screening potentially immunotoxic contaminants as single compounds or as complex mixtures. Compared to the reference site, filter-sterilized Atlantic Wood sediment extracts (water accommodated fractions) induced nitric oxide and IL-6 secretion as well as inducible nitric oxide synthase and COX-2 proteins at levels comparable to or higher than those induced by LPS treatments alone. Extracts also increased phagocytic activity by macrophages. Using a limulus lysate assay, we show that bacterial endotoxin levels in Atlantic Wood extracts are higher than in reference extracts and that polymyxin-B chelation ameliorates proinflammatory effects. These findings illuminate the reality of sediment constituents other than toxic compounds previously associated with developmental abnormalities and carcinogenesis in killifish from the Atlantic Wood site. Perhaps these data also suggest the presence of contaminant-adapted consortia of sediment microbes at many heavily polluted sites worldwide compared to less contaminated sites. Environ Toxicol Chem 2021;40:1576-1585. © 2021 SETAC.

Background Meloidae (blister beetles) are known to synthetize cantharidin (CA), a toxic and defensive terpene mainly stored in male accessory glands (MAG) and emitted outward through reflex-bleeding. Recent progresses in understanding CA biosynthesis and production organ(s) in Meloidae have been made, but the way in which self-protection is achieved from the hazardous accumulation and release of CA in blister beetles has been experimentally neglected. To provide hints on this pending question, a comparative de novo assembly transcriptomic approach was performed by targeting two tissues where CA is largely accumulated and regularly circulates in Meloidae: the male reproductive tract (MRT) and the haemolymph. Differential gene expression profiles in these tissues were examined in two blister beetle species, Lydus trimaculatus (Fabricius, 1775) (tribe Lyttini) and Mylabris variabilis (Pallas, 1781) (tribe Mylabrini). Upregulated transcripts were compared between the two species to identify conserved genes possibly involved in CA detoxification and transport. Results Based on our results, we hypothesize that, to avoid auto-intoxication, ABC, MFS or other solute transporters might sequester purported glycosylated CA precursors into MAG, and lipocalins could bind CA and mitigate its reactivity when released into the haemolymph during the autohaemorrhaging response. We also found an over-representation in haemolymph of protein-domains related to coagulation and integument repairing mechanisms that likely reflects the need to limit fluid loss during reflex-bleeding. Conclusions The de novo assembled transcriptomes of L. trimaculatus and M. variabilis here provided represent valuable genetic resources to further explore the mechanisms employed to cope with toxicity of CA in blister beetle tissues. These, if revealed, might help conceiving safe and effective drug-delivery approaches to enhance the use of CA in medicine.

Chronic wound management becomes a complex procedure because of the persistence of forming biofilm pathogens that do not respond to antimicrobial treatment. The aim of this paper is to detect the Graphene Oxide-GO effect on Staphylococcus aureus and Pseudomonas aeruginosa dual species wound biofilm in Lubbock Chronic Wound Biofilm-LCWB model. LCWB is a recognized model that mimics the spatial microbial colonization into chronic wounds and reproduces the wound and its clot. Staphylococcus aureus PECHA 10 and P. aeruginosa PECHA 4, are the pathogens used in the study. The GO effect on both in forming and mature biofilms, is detected by the evaluation of the CFU/mg reduction, the cell viability and ultrastructural analysis of the treated LCWBs. Graphene Oxide, at 50 mg/l, shows a significant antibiofilm effect in forming and mature LCWBs. In particular, during the biofilm formation, GO reduces the S. aureus and P. aeruginosa growth of 55.05% ± 4.73 and 44.18% ± 3.91 compared to the control. In mature biofilm, GO affects S. aureus and P. aeruginosa by reducing their growth of 70.24% ± 4.47 and 63.68% ± 17.56, respectively. Images taken by SEM show that GO display a disaggregated microbial effect also disrupting the fibrin network of the wound-like biofilm framework. In conclusion, GO used against microorganisms grown in LCWB, displays a significant inhibitory action resulting in a promising tool for potential application in wound management.

Emerging evidence suggests the role of environmental chemicals, in particular endocrine-disrupting chemicals (EDCs), in progression of breast cancer and treatment resistance, which can impact survival outcomes. However, most research tends to focus on tumor etiology and the effect of single chemicals, offering little insight into the effects of realistic complex mixture exposures on tumor progression. Herein, we investigated the effect of a polycyclic aromatic hydrocarbon (PAH)-enriched EDC mixture in a panel of normal and breast cancer cells and in a tumor organoid model. Cells or organoids in culture were treated with EDC mixture at doses estimated from US adult intake of the top four PAH compounds within the mixture from the National Health and Nutrition Examination Survey database. We demonstrate that low-dose PAH mixture (6, 30 and 300 nM) increased aryl hydrocarbon receptor (AhR) expression and CYP activity in estrogen receptor (ER) positive but not normal mammary or ER-negative breast cancer cells, and that upregulated AhR signaling corresponded with increased cell proliferation and expression of antiapoptotic and antioxidant proteins XIAP and SOD1. We employed a mathematical model to validate PAH-mediated increases in AhR and XIAP expression in the MCF-7 ER-positive cell line. Furthermore, the PAH mixture caused significant growth increases in ER-negative breast cancer cell derived 3D tumor organoids, providing further evidence for the role of a natural-derived PAH mixture in enhancing a tumor proliferative phenotype. Together, our integrated cell signaling, computational and phenotype analysis reveals the underlying mechanisms of EDC mixtures in breast cancer progression and survival.

Objectives The present study aims to verify the relationship between glucose consumption and uptake of 18F-2-deoxy-glucose (FDG) in the skeletal muscle (SM) of experimental models of streptozotocin-induced diabetes mellitus (STZ-DM). Methods The study included 36 Balb/c mice. Two weeks after intraperitoneal administration of saline (control group, n = 18) or 150 mg streptozotocin (STZ-DM group, n = 18), the two cohorts were submitted to an oral glucose tolerance test and were further subdivided into three groups (n = 6 each): untreated and treated with metformin (MTF) at low or high doses (10 or 750 mg/kg daily, respectively). Two weeks thereafter, all mice were submitted to dynamic micro–positron emission tomography (PET) imaging after prolonged fasting. After sacrifice, enzymatic pathways and response to oxidative stress were evaluated in harvested SM. Results On PET imaging, the FDG uptake rate in hindlimb SM was significantly lower in nondiabetic mice as compared with STZ-DM–untreated mice. MTF had no significant effect on SM FDG uptake in untreated mice; however, its high dose induced a significant decrease in STZ-DM animals. Upon conventional analysis, the SM standard uptake value was higher in STZ-DM mice, while MTF was virtually ineffective in either control or STZ-DM models. This metabolic reprogramming was not explained by any change in cytosolic glucose metabolism. By contrast, it closely agreed with the catalytic function of hexose-6P-dehydrogenase (H6PD; i.e., the trigger of a specific pentose phosphate pathway selectively located within the endoplasmic reticulum). In agreement with this role, the H6PD enzymatic response to both STZ-DM and MTF matched the activation of the NADPH-dependent antioxidant responses to the increased generation of reactive oxygen species caused by chronic hyperglycemia. Ex vivo analysis of tracer kinetics confirmed that the enhanced SM avidity for FDG occurred despite a significant reduction in glucose consumption, while it was associated with increased radioactivity transfer to the endoplasmic reticulum. Conclusions These data challenge the current dogma linking FDG uptake to the glycolytic rate. They instead introduce a new model considering a strict link between the uptake of this glucose analog, H6PD reticular activity, and oxidative damage in diabetes, at least under fasting condition., Highlights • The accepted link between glucose consumption and uptake of hexose analogs such as FDG and 2DG is looser than expected. • FDG selectively tracks the activity of a pentose-phosphate shunt triggered by H6PD. • FDG uptake in the skeletal muscles tracks the hyperglycemia-related oxidative damage. • FDG uptake might represent an index of NADPH renewal indicating the ER-PPP response to oxidative stress.

New bio-based polymers used as consolidants for waterlogged archaeological wood were studied and their performance on real wooden samples was assessed. In particular, three oligoamides (oligo ethylene- l -tartaramide, oligo esamethylene- l -tartaramide and copolymer between ethylenediamine, adipic and tartaric acids) and allyl α,α′-trehalose/vinyl alcohol (ATR/VOH) copolymer were selected as consolidants. The affinity for wood and the reversibility of the treatments were evaluated together with certain physical properties of treated wood, i.e. maximum water content (MWC), basic density (ρb) and volumetric shrinkage (βv). A specific diagnostic protocol was optimized in order to obtain reproducible and reliable results. A good affinity for a degraded wood rich in lignin was demonstrated for all tested consolidants. For what concerns the reversibility of the treatments, the oligoamides showed higher reversibility than the ATR/VOH copolymer. The penetration ability was good for all selected products. Good results in terms of MWC reduction and ρb increase were obtained with the oligo esamethylene- L -tartaramide, the oligo ethylene- L -tartaramide and the ATR/VOH copolymer.

I respond to criticisms raised by Kim et al. (2018) to my model concerning the origin of the tRNA molecule. In particular, their model would hypothesize the tRNA originated due to the ligation of three hairpin structures followed by two deletions, while my model predicts that this molecule derived from the assembly of only two hairpin-like structures. Thus, using the Ockham razor, the latter model would be chosen because it required fewer hypotheses. Furthermore, the predictions on homology between the different regions of the tRNA molecule as predicted by my model would be statistically more significant than those predicted by their model. Moreover, it would be above all the existence of molecular fossils - i.e. the split tRNA genes - to corroborate the model of the assembly of only two hairpins. These fossils would be completely absent from the Kim et al. (2018) model.

BackgroundThe long noncoding RNA LIPCAR (Long Intergenic noncoding RNA Predicting CARdiac remodeling) has emerged as a promising biomarker in cardiac disease and cardiac remodeling. To determine whether LIPCAR levels help for a molecular phenotyping of chronic heart failure (HF) patients, this study assessed the association of LIPCAR with severity of the disease and its progression, and with risk of death or hospitalization in HF patients.MethodsLIPCAR was measured in plasma of 967 HF patients with symptomatic heart failure participating in the Gruppo Italiano per lo Studio della Sopravvivenza nell'Insufficienza Cardiaca - Heart Failure (GISSI-HF) biohumoral sub-study.ResultsPlasma levels of LIPCAR were significantly associated with functional impairment as assessed by the New York Heart Association (NYHA) class, kidney function as reflected by estimated glomerular filtration rate, and creatinine, hemoglobin and mitral insufficiency. In females, these associations were more marked as compared to males. LIPCAR plasma levels were significantly related to the two cardiac markers, N-terminal pro-B type natriuretic peptide and high-sensitivity cardiac troponin T, but not to inflammatory markers such as high sensitivity C-reactive protein and pentraxin-3, nor to patient reported outcomes such as depression and quality of life. HF patients with high LIPCAR levels univariately showed significantly higher incidence of cardiovascular hospitalizations but not of death; after adjusting for covariates, no significant effects of LIPCAR were found for cardiovascular hospitalizations.ConclusionThe circulating long noncoding RNA LIPCAR was increased in HF patients with higher NYHA class, impaired kidney function, and lower hemoglobin, which are indicators of patients’ overall state.

Copper (Cu) dyshomeostasis plays a pivotal role in several neuropathologies, such as Parkinson’s disease (PD). Metal accumulation in the central nervous system (CNS) could result in loss-of-function of proteins involved in Cu metabolism and redox cycling, generating reactive oxygen species (ROS). Moreover, neurodegenerative disorders imply the presence of an excess of misfolded proteins known to lead to neuronal damage. In PD, Cu accumulates in the brain, binds α-synuclein, and initiates its aggregation. We assessed the correlation between neuronal differentiation, Cu homeostasis regulation, and α-synuclein phosphorylation. At this purpose, we used differentiated SHSY5Y neuroblastoma cells to reproduce some of the characteristics of the dopaminergic neurons. Here, we reported that differentiated cells expressed a significantly higher amount of a copper transporter protein 1 (CTR1), increasing the copper uptake. Cells also showed a significantly more phosphorylated form of α-synuclein, further increased by copper treatment, without modifications in α-synuclein levels. This effect depended on the upregulation of the polo-like kinase 2 (PLK2), whereas the levels of the relative protein phosphatase 2A (PP2A) remained unvaried. No changes in the oxidative state of the cells were identified. The Cu dependent alteration of α-synuclein phosphorylation pattern might potentially offer new opportunities for clinical intervention.

PM10 samples were collected in a site of Apulia region (South-East Italy) near a steel plant when the winds were blowing from the NW and the SE to exclude and include, respectively, the contribution of the industrial source in order to study the aerosol particles in detail and attribute them to the most appropriate emission sources. Size, morphology and chemical composition of 6500 individual particles, with an equivalent spherical diameter (ESD) between 0.4 ˗ 1 μm, 1 ˗ 2.5 μm and 2.5 ˗ 10 μm, were analysed by scanning electron microscopy coupled with energy dispersive X-ray analysis (SEM - EDX). These particles were classified in the thirteen groups: aluminosilicates, silicates, iron oxides, metal oxides, iron mixtures, aluminosilicates with sulphur, calcium sulphates, silicate-sulphate mixed particles, carbonates, carbonates-silicates, soot, biological particles and carbon rich particles. Source apportionment analyses were carried out separately for the three dimensional ranges considered and for the two wind directions from North-West and South-East and nine main sources were distinguished. The soil and the soot components are higher when the wind blow from the NW than the SE, while the anthropogenic sources are evident with the wind from the SE due to the presence of iron and carbonaceous particles characterized by typical spherical shapes and chemical elements as Ca, Na and K which could to be indicative of industrial activity. The single particle analysis let better identify the emission sources that persist on the territory, attributing to them the particles characterized by peculiar morphology and chemical composition.

Herein we report the electropolymerization of a scopoletin based molecularly imprinted polymer (MIP) for the detection of lysozyme (Lyz), an enzymatic marker of several diseases in mammalian species. Two different approaches have been used for the imprinting of lysozyme based, respectively, on the use of a monomer-template mixture and on the covalent immobilization of the enzyme prior to polymer synthesis. In the latter case, a multi-step protocol has been exploited with preliminary functionalization of gold electrode with amino groups, via 4-aminothiophenol, followed by reaction with glutaraldehyde, to provide a suitable linker for lysozyme. Each step of surface electrode modification has been followed by cyclic voltammetry and electrochemical impedance spectroscopy, which has been also employed to test the electrochemical responses of the developed MIP. The sensors show good selectivity to Lyz and detect the enzyme at concentrations up to 292 mg/L (20 &mu, M), but with different performances, depending on the used imprinting approach. An imprinting factor equal to 7.1 and 2.5 and a limit of detection of 0.9 mg/L (62 nM) and 2.1 mg/L (141 nM) have been estimated for MIPs prepared with and without enzyme immobilization, respectively. Competitive rebinding experiment results show that this sensing material is selective for Lyz determination. Tests were performed using synthetic saliva to evaluate the potential application of the sensors in real matrices for clinical purposes.

In the history of diagnostics, breath analysis was one of the first method used until the breakthrough of biochemical testing technology. Today, breath analysis has made a comeback with the development of gas analyzers and e‐noses, demonstrating its power in its applicability for diagnosing a wide range of diseases. The physical basis of multiple chemical sensitivity (MCS), an emerging environmental disease, is difficult to understand because it is based on the scenario of chronic hypoxia, with a complex of chemical compounds that trigger the syndrome and result in multiple symptoms. The aim of this study was to investigate MCS by analyzing exhaled volatile organic compounds (VOCs). The volatile, metabolic picture could be a putative gold standard for understanding and diagnosing the disease. The study was based on recording in resting condition using the noninvasive passive e‐nose contactless breath test, the Olfactory Real‐Time Volatile Organic Compounds (ORT‐VOC) test in MCS, and control samples. The VOCs profile distinguished between disease and health. It also distinguished the gender‐related volatile profile with significant robustness. The results trace a putative compensatory physiological pathway elicited by increased lactate, leading to acidosis, and hyperventilation, resulting in the production of specific VOCs. We conclude that breath testing is a valuable tool to investigate the hypoxia‐related VOC profile, facilitating MCS diagnosis.

Chemically skinned fibres allow the study of human muscle contractile function in vitro. A particularly important parameter is specific force (SF), that is, maximal isometric force divided by cross-sectional area, representing contractile quality. Although SF varies substantially between studies, the magnitude and cause of this variability remains puzzling. Here, we aimed to summarize and explore the cause of variability in SF between studies. A systematic search was conducted in Medline, Embase and Web of Science databases in June 2020, yielding 137 data sets from 61 publications which studied healthy, young adults. Five-fold differences in mean SF data were observed. Adjustments to the reported data for key methodological differences allowed between-study comparisons to be made. However, adjustment for fibre shape, swelling and sarcomere length failed to significantly reduce SF variance (I2 = 96%). Interestingly, grouping papers based on shared authorship did reveal consistency within research groups. In addition, lower SF was found to be associated with higher phosphocreatine concentrations in the fibre activating solution and with Triton X-100 being used as a skinning agent. Although the analysis showed variance across the literature, the ratio of SF in single fibres containing myosin heavy chain isoforms IIA or I was found to be consistent across research groups. In conclusion, whilst the skinned fibre technique is reliable for studying in vitro force generation of single fibres, the composition of the solution used to activate fibres, which differs between research groups, is likely to heavily influence SF values.

We used the Moran’s I index of global spatial autocorrelation with the aim of studying the distribution of the physicochemical or biological properties of amino acids within the genetic code table. First, using this index we are able to identify the amino acid property - among the 530 analyzed - that best correlates with the organization of the genetic code in the set of amino acid permutation codes. Considering, then, a model suggested by the coevolution theory of the genetic code origin - which in addition to the biosynthetic relationships between amino acids took into account also their physicochemical properties - we investigated the level of optimization achieved by these properties either on the entire genetic code table, or only on its columns or only on its rows. Specifically, we estimated the optimization achieved in the restricted set of amino acid permutation codes subject to the constraints derived from the biosynthetic classes of amino acids, in which we identify the most optimized amino acid property among all those present in the database. Unlike what has been claimed in the literature, it would appear that it was not the polarity of amino acids that structured the genetic code, but that it could have been their partition energy instead. In actual fact, it would seem to reach an optimization level of about 96% on the whole table of the genetic code and 98% on its columns. Given that this result has been obtained for amino acid permutation codes subject to biosynthetic constraints, that is to say, for a model of the genetic code consistent with the coevolution theory, we should consider the following conclusions reasonable. (i) The coevolution theory might be corroborated by these observations because the model used referred to the biosynthetic relationships between amino acids, which are suggested by this theory as having been fundamental in structuring the genetic code. (ii) The very high optimization on the columns of the genetic code would not only be compatible but would further corroborate the coevolution theory because this suggests that, as the genetic code was structured along its rows by the biosynthetic relationships of amino acids, on its columns strong selective pressure might have been put in place to minimize, for example, the deleterious effects of translation errors. (iii) The finding that partition energy could be the most optimized property of amino acids in the genetic code would in turn be consistent with one of the main predictions of the coevolution theory. In other words, since the partition energy is reflective of the protein structure and therefore of the enzymatic catalysis, the latter might really have been the main selective pressure that would have promoted the origin of the genetic code. Indeed, we observe that the β-strands show an optimization percentage of 94.45%, so it is possible to hypothesize that they might have become the object of selection during the origin of the genetic code, conditioning the choice of biosynthetic relationships between amino acids. (iv) The finding that the polarity of amino acids is less optimized than their partition energy in the genetic code table might be interpreted against the physicochemical theories of the origin of the genetic code because these would suggest, for example, that a very high optimization of the polarity of amino acids in the code could be an expression of interactions between amino acids and codons or anticodons, which would have promoted their origin. This might now become less sustainable, given the very high optimization that is instead observed in favor of partition energy but not polarity. Finally, (v) the very high optimization of the partition energy of amino acids would seem to make a neutral origin of the ability of the genetic code to buffer, for example, the deleterious effects of translation errors very unlikely. Indeed, an optimization of about 100% would seem that it might not have been achieved by a simple neutral process, but this ability should probably have been generated instead by the intervention of natural selection. In actual fact, we show that the neutral hypothesis of the origin of error minimization has been falsified for the model analyzed here. Therefore, we will discuss our observations within the theories proposed to explain the origin of the organization of the genetic code, reaching the conclusion that the coevolution theory is the most strongly corroborated theory.

Blister beetles owe their name to their ability to release cantharidin, a blistering terpene, the highest concentration of which is retained in male accessory glands. The anatomy and ultrastructure of the three pairs of male reproductive accessory glands and the glandular region of the two vasa deferentia of Meloe proscarabaeus were investigated using light, electron and ion beam microscopy. All of the mesodermal glands here analysed share a common structural organization with an outer muscular layer and an inner glandular epithelium facing a broad lumen in which the secretory products are released. Developed rough endoplasmic reticulum, Golgi systems, abundant mitochondria, numerous secretory vesicles and a microvillated apical membrane are commonly found in the cells of different glandular epithelia, suggesting that all accessory gland pairs as well as the vasa deferentia are involved in an active synthesis. Nevertheless, each pair of glands appears specialized in the production of a specific set of substances, as suggested by the peculiarities in cellular ultrastructure and by the different aspect of the secretions stored in their glandular lumen. The above cited features of male accessory glands of M. proscarabaeus are compared with those of other beetles and some hints on their potential role in producing and/or concentrating cantharidin are provided.

The main purpose of this study was to explain the internal fine structure of potential antennal thermo- and hygroreceptive sensilla, their innervation specifics, and responses of the sensory neurons to thermal and humidity stimuli in an elaterid beetle using focused ion beam scanning electron microscopy and electrophysiology, respectively. Several essential, high temperature induced turning points in the locomotion were determined using automated video tracking. Our results showed that the sensilla under study, morphologically, are identical to the dome-shaped sensilla (DSS) of carabids. A cold-hot neuron and two bimodal hygro-thermoreceptor neurons, the moist-hot and dry-hot neuron, innervate them. Above 25–30 °C, all the three neurons, at different threshold temperatures, switch from regular spiking to temperature dependent spike bursting. The percentage of bursty DSS neurons on the antenna increases with temperature increase suggesting that this parameter of the neurons may encode noxious heat in a graded manner. Thus, we show that besides carabid beetles, elaterids are another large group of insects with this ability. The threshold temperature of the beetles for onset of elevated locomotor activity (OELA) was lower by 11.9 °C compared to that of critical thermal maximum (39.4 °C). Total paralysis occurred at 41.8 °C. The threshold temperatures for spike bursting of the sensory neurons in DSS and OELA of the beetles coincide suggesting that probably the spike bursts are responsible for encoding noxious heat when confronted. In behavioural thermoregulation, spike bursting DSS neurons serve as a fast and firm three-fold early warning system for the beetles to avoid overheating and death.

Background. The so-called “vaccine hesitancy” still represents a common phenomenon that undermines the effectiveness of vaccination campaigns. In 2020, the Italian Medicines Agency recommended to bring forward the flu vaccination campaign, whose importance was also emphasized for patients with Multiple Sclerosis (MS). We aimed to assess vaccination behavior in patients with MS to prepare for the upcoming SARS-CoV-2 vaccination challenge. Methods. This is an observational study carried out in one MS clinical Centre that enrolled all MS patients who were eligible for any of the flu vaccines recommended by the Italian medicines Agency. Results. 194 patients were enrolled. Patients’ mean age was 43.9 years and 66% were female. Comorbidities, mainly represented by non-autoimmune diseases, were identified in 52% of patients. Almost all patients were receiving a DMT during the study period, mainly dimethyl fumarate, natalizumab, teriflunomide, and interferon. Out of 194 patients, 58.2% accepted to be vaccinated. No statistically significant differences were found, except for the use of natalizumab, which was higher among vaccinated patients. Conclusion. The results of our study emphasize the importance of education and communication campaigns addressed both to healthcare providers and patients with MS, especially considering that MS patients are currently receiving COVID-19 vaccinations.

Sites along the Elizabeth River are contaminated with polycyclic aromatic hydrocarbons (PAHs) from historical creosote production and other industrial processes. Previous studies have demonstrated that Atlantic killifish collected from sites throughout the Elizabeth River display resistance to the teratogenic effects of PAH-exposure in a manner commensurate with sediment PAH concentrations. The current study characterized various chemical pollutants in sediment and investigated the effects of aqueous sediment extracts from sites along the Elizabeth River to the cardiac development of Atlantic killifish embryos from fish collected from an uncontaminated reference site. Embryonic cardiac deformities were more prevalent after exposure to extracts from sites with high PAH loads. However, activation of cytochrome P4501A, a gene up-regulated by PAH-induction of the aryl hydrocarbon receptor and measured using an in ovo EROD assay, did not consistently increase with PAH concentrations. This work further characterizes sediments in the Elizabeth River, as well as provides insight into the evolutionary pressures at each ER site.

Aims. The study investigated the effect of high-intensity interval training in hypoxia and normoxia on serum concentrations of proangiogenic factors, nitric oxide, and inflammatory responses in healthy male volunteers. Methods. Twelve physically active male subjects completed a high-intensity interval training (HIIT) in normoxia (NorTr) and in normobaric hypoxia (HypTr) (FiO2 = 15.2%). The effects of HIIT in hypoxia and normoxia on maximal oxygen uptake, hypoxia-inducible factor-1-alpha, vascular endothelial growth factor, nitric oxide, and cytokines were analyzed. Results. HIIT in hypoxia significantly increases maximal oxygen uptake (p=0.01) levels compared to pretraining levels. Serum hypoxia-inducible factor-1 (p=0.01) and nitric oxide levels (p=0.05), vascular endothelial growth factor (p=0.04), and transforming growth factor-β (p=0.01) levels were increased in response to exercise test after hypoxic training. There was no effect of training conditions for serum baseline angiogenic factors and cytokines (p>0.05) with higher HIF-1α and NO levels after hypoxic training compared to normoxic training (F = 9.1; p<0.01 and F = 5.7; p<0.05, respectively). Conclusions. High-intensity interval training in hypoxia seems to induce beneficial adaptations to exercise mediated via a significant increase in the serum concentrations of proangiogenic factors and serum nitric oxide levels compared to the same training regimen in normoxia.

The anionic surfactant sodium lauryl ether sulphate (SLES) is the main component in most foaming agents used for mechanized tunneling excavation. The process produces huge amounts of soil debris that can have a potential impact on ecosystems. The lack of accurate information about SLES persistence in excavated soil has aroused increasing concern about how it is recycled. The objective of this study was to assess SLES biodegradability in two commercial foaming agents (P1 and P2). Microcosm experiments were performed with two different soils collected from a tunnel construction site and conditioned with P1 or P2 (85.0 or 83.0 mg kg −1 of SLES, respectively). At selected times soil samples were collected for assessing the SLES residual concentration using Pressured Liquid Extraction followed by methylene blue active substance analysis (MBAS). Simultaneously, soil microbial abundance (DAPI counts), viability (Live/Dead method), activity (dehydrogenase analysis) and phylogenetic structure (Fluorescent In Situ Hybridization) were evaluated. SLES halved faster in the silty-clay soil (6 d) than in the gravel in a clay-silty-sand matrix (8–9 days). At day 28 it was degraded in both soils. Its biodegradation was ascribed to the significant increase in Gamma-Proteobacteria. At this time, the spoil material can be considered as a by-product.

Plastics are ubiquitous anthropogenic contaminants that are a growing concern in aquatic environments. The ecological implications of macroplastics pollution are well documented, but less is known about nanoplastics. The current study investigates the potential adverse effects of nanoplastics, which likely contribute to the ecological burden of plastic pollution. To this end, we examined whether a dietary exposure of adult zebrafish (Danio rerio) to polystyrene nanoparticles (PS NPs) could lead to the transfer of nanoplastics to the offspring, and whether nanoplastics exposure affects zebrafish physiology. Specifically, adult female and male zebrafish (F0 generation) were exposed to PS NPs via diet for one week and bred to produce the F1 generation. Four F1 groups were generated: control (unexposed females and males), maternal (exposed females), paternal (exposed males), and co-parental (exposed males and females). Co-parental PS NP exposure did not significantly affect reproductive success. Assessment of tissues from F0 fish revealed that exposure to PS NPs significantly reduced glutathione reductase activity in brain, muscle, and testes, but did not affect mitochondrial function parameters in heart or gonads. Assessment of F1 embryos and larvae revealed that PS NPs were present in the yolk sac, gastrointestinal tract, liver, and pancreas of the maternally and co-parentally exposed F1 embryos/larvae. Bradycardia was also observed in embryos from maternal and co-parental exposure groups. In addition, the activity of glutathione reductase and the levels of thiols were reduced in F1 embryos/larvae from maternal and/or co-parental exposure groups. Mitochondrial function and locomotor activity were not affected in F1 larvae. This study demonstrates that (i) PS NPs are transferred from mothers to offspring, and (ii) exposure to PS NPs modifies the antioxidant system in adult tissues and F1 larvae. We conclude that PS NPs could bioaccumulate and be passed on to the offspring, but this does not lead to major physiological disturbances.

I analyse the origin of the genetic code in the light of the evolution of biological catalysts. I discuss the rudimentary forms that the genetic code assumed in the presence of a catalysis performed by ions or by low molecular weight molecules, such as nucleotide coenzymes. However, it is only with the advent of a mixed polymer made of RNA and peptides - covalently linked - that the genetic code took on a clearer form. Indeed, the first true form of coding appeared. Furthermore, interacting peptidated RNAs promoted an extremely rudimentary form of protein synthesis. This stage evolved into a stage in which proto-mRNAs guided interactions among peptidated RNAs aimed at the synthesis of peptidated RNAs having an active catalytic function. Finally, the invasion of aminoacylated proto-tRNAs with specific amino acids, coming from amino acid metabolism, and recognising only three bases on these proto-mRNAs with reading frames larger than three bases, would have triggered the birth of actual mRNAs, i.e. the origin of codons. All this would have linked the metabolism of amino acids to the origin of mRNAs and therefore to the origin of the organization of the genetic code, as maintained by the coevolution theory of the genetic code.

Organophosphate (OP) compounds comprise one of the most widely used classes of insecticides worldwide. OPs have been shown to have negative human health impacts, particularly developmental neurotoxicity. However, neurotoxic impacts in later adulthood and during the aging process are relatively uncharacterized. The present study examined diazinon (DZN), an OP, to determine the neurobehavioral consequences, in addition to mitochondrial dysfunction on a macroscale (whole organism basal respiration) and on a microscale (whole organ mitochondrial respiration), using zebrafish (ZF) as a model. One group of 14-month-old adult ZF were exposed acutely as adults (0.4, 1.25, and 4.0 μM) for five days and tested as adults, and another group was exposed developmentally 5–120 h post-fertilization (70, 210, and 700 nM) and tested at larval, adolescent, adult, and aging life stages. ZF exposed acutely as adults did not display many significant neurobehavioral impacts or mitochondrial dysfunction. Conversely, the embryonically exposed ZF showed altered behavioral functions at each stage of life which emerged and attenuated as fish transitioned from each developmental stage to the next. Mitochondrial oxygen consumptions measurement results for developmentally DZN exposed ZF showed significant increases in the low and middle dose groups in organs such as the brain and testes. Overall, there is an indication that early developmental exposure to DZN had continuing adverse neurobehavioral and cellular consequences throughout their lives well into adulthood and aging periods.

MYCN drives aggressive behavior and refractoriness to chemotherapy, in several tumors. Since MYCN inactivation in clinical settings is not achievable, alternative vulnerabilities of MYCN-driven tumors need to be explored to identify more effective and less toxic therapies. We previously demonstrated that PARP inhibitors enhance MYCN-induced replication stress and promote mitotic catastrophe, counteracted by CHK1. Here, we showed that PARP and CHK1 inhibitors synergized to induce death in neuroblastoma cells and in primary cultures of SHH-dependent medulloblastoma, their combination being more effective in MYCN amplified and MYCN overexpressing cells compared to MYCN non-amplified cells. Although the MYCN amplified IMR-32 cell line carrying the p.Val2716Ala ATM mutation showed the highest sensitivity to the drug combination, this was not related to ATM status, as indicated by CRISPR/Cas9-based correction of the mutation. Suboptimal doses of the CHK1 inhibitor MK-8776 plus the PARP inhibitor olaparib led to a MYCN-dependent accumulation of DNA damage and cell death in vitro and significantly reduced the growth of four in vivo models of MYCN-driven tumors, without major toxicities. Our data highlight the combination of PARP and CHK1 inhibitors as a new potential chemo-free strategy to treat MYCN-driven tumors, which might be promptly translated into clinical trials.

Background: An unbalanced skin microbiota due to an increase in pathogenic vs. commensal bacteria can be efficiently tackled by using prebiotics. The aim of this work was to identify novel prebiotic combinations by exerting species-specific action between S. aureus and S. epidermidis strains. Methods: First, the antimicrobial/antibiofilm effect of Xylitol-XYL and Galacto-OligoSaccharides&ndash, GOS combined with each other at different concentrations (1, 2.5, 5%) against S. aureus and S. epidermidis clinical strains was evaluated in time. Second, the most species-specific concentration was used to combine XYL with Fructo-OligoSaccharides&ndash, FOS, IsoMalto-Oligosaccharides&ndash, IMO, ArabinoGaLactan&ndash, LAG, inulin, dextran. Experiments were performed by OD600 detection, biomass quantification and LIVE/DEAD staining. Results: 1% XYL + 1% GOS showed the best species-specific action with an immediate antibacterial/antibiofilm action against S. aureus strains (up to 34.54% &plusmn, 5.35/64.68% &plusmn, 4.77) without a relevant effect on S. epidermidis. Among the other prebiotic formulations, 1% XYL plus 1% FOS (up to 49.17% &plusmn, 21.46/37.59% &plusmn, 6.34) or 1% IMO (up to 41.28% &plusmn, 4.88/36.70% &plusmn, 10.03) or 1% LAG (up to 38.21% &plusmn, 5.31/83.06% &plusmn, 5.11) showed antimicrobial/antibiofilm effects similar to 1% XYL+1% GOS. For all tested formulations, a prevalent bacteriostatic effect in the planktonic phase and a general reduction of S. aureus biofilm formation without loss of viability were recorded. Conclusion: The combinations of 1% XYL with 1% GOS or 1% FOS or 1% IMO or 1% LAG may help to control the balance of skin microbiota, representing good candidates for topic formulations.

It has been firmly observed that replicative DNA polymerases of bacteria, archaea and eukaryotes are not homologous proteins. This lack of homology in the replication apparatus among the domains of life is not only compatible with but would seem to imply the view that the emergence of DNA occurred in the fundamental cellular lineages. In consequence, this diversity of DNA polymerase would go back to the level of ancestors of the domains of life and to the evolutionary time in which the DNA emerged. Therefore, the presumed evolutionary stage linked to the RNA- > DNA transition would have occurred only at the level of ancestors of the main lineages of the tree of life. Thus, the high noise associated with this major evolutionary transition and the impossibility for a cellular stage to generate different fundamental genetically profound traits - such as the different replication apparatuses of bacteria, archaea and eukaryotes - would imply not only that the last universal common ancestor (LUCA) was a progenote but that the ancestors of the domains of life were also at this evolutionary stage. So, I criticize the hypotheses which want, instead, that completely different cells - such as, bacteria and archaea - could have originated from a cellular LUCA.

Plastics are world-wide pollutants that pose a potential threat to wildlife and human health. Small plastic particles, such as microplastics and nanoplastics, are easily ingested, and can act as a Trojan Horse by carrying microorganisms and pollutants. This study investigated the potential role of the Trojan Horse effect in the toxicity of nanoplastics to the vertebrate model organism, zebrafish (Danio rerio). First, we investigated if this effect could affect the toxicity of nanoplastics. Second, we analyzed if it could contribute to the biodistribution of the associated contaminants. And third, we focused on its effect on the mitochondrial toxicity of nanoplastics. We incubated 44 nm polystyrene nanoparticles with a real-world mixture of polycyclic aromatic hydrocarbons (PAHs) for 7 days and removed the free PAHs by ultrafiltration. We dosed embryos with 1 ppm of nanoplastics (NanoPS) or PAH-sorbed nanoplastics (PAH-NanoPS). Neither type of plastic particle caused changes in embryonic and larval development. Fluorescence microscopy and increased EROD activity suggested the uptake of PAHs in larvae exposed to PAH-NanoPS. This coincided with higher concentrations in the yolk sac and the brain. However, PAH-only exposure leads to their accumulation in the yolk sac but not in the brain, suggesting that that the spatial distribution of bioaccumulated PAHs can differ depending on their source of exposure. Both nanoplastic particles affected mitochondrial energy metabolism but caused different adverse effects. While NanoPS decreased NADH production, PAH-NanoPS decreased mitochondrial coupling efficiency and spare respiratory capacity. In summary, the addition of PAHs to the surface of nanoplastics did not translate into increased developmental toxicity. Low levels of PAHs were accumulated in the organisms, and the transfer of PAHs seems to happen in tissues and possibly organelles where nanoplastics accumulate. Disruption of the energy metabolism in the mitochondria may be a key factor in the toxicity of nanoplastics, and the Trojan Horse effect may amplify this effect.

The SHiP experiment is proposed to search for very weakly interacting particles beyond the Standard Model which are produced in a 400 GeV/$c$ proton beam dump at the CERN SPS. About $10^{11}$ muons per spill will be produced in the dump. To design the experiment such that the muon-induced background is minimized, a precise knowledge of the muon spectrum is required. To validate the muon flux generated by our Pythia and GEANT4 based Monte Carlo simulation (FairShip), we have measured the muon flux emanating from a SHiP-like target at the SPS. This target, consisting of 13 interaction lengths of slabs of molybdenum and tungsten, followed by a 2.4 m iron hadron absorber was placed in the H4 400 GeV/$c$ proton beam line. To identify muons and to measure the momentum spectrum, a spectrometer instrumented with drift tubes and a muon tagger were used. During a three-week period a dataset for analysis corresponding to $(3.27\pm0.07)~\times~10^{11}$ protons on target was recorded. This amounts to approximatively 1% of a SHiP spill. The SHiP experiment will search for very weakly interacting particles beyond the Standard Model which are produced in a 400 \GeV/$c$ proton beam dump at the CERN SPS. About $10^{11}$ muons per spill will be produced in the dump. To design the experiment such that the muon-induced background is minimized, a precise knowledge of the muon spectrum is required. To validate the muon flux generated by our Pythia and GEANT4 based Monte Carlo simulation (FairShip), we have measured the muon flux emanating from a SHiP-like target at the SPS. This target, consisting of 13 interaction lengths of slabs of molybdenum and tungsten, followed by a 2.4 m iron hadron absorber was placed in the H4 400~\GeV/$c$ proton beam line. To identify muons and to measure the momentum spectrum, a spectrometer instrumented with drift tubes and a muon tagger were used. During a three-week period a dataset for analysis corresponding to $(3.27\pm0.07)~\times~10^{11}$ protons on target was recorded. This amounts to approximatively 1\% of a SHiP spill. The SHiP experiment is proposed to search for very weakly interacting particles beyond the Standard Model which are produced in a 400 GeV/c proton beam dump at the CERN SPS. About $10^{11}$ muons per spill will be produced in the dump. To design the experiment such that the muon-induced background is minimized, a precise knowledge of the muon spectrum is required. To validate the muon flux generated by our Pythia and GEANT4 based Monte Carlo simulation (FairShip), we have measured the muon flux emanating from a SHiP-like target at the SPS. This target, consisting of 13 interaction lengths of slabs of molybdenum and tungsten, followed by a 2.4 m iron hadron absorber was placed in the H4 400 GeV/c proton beam line. To identify muons and to measure the momentum spectrum, a spectrometer instrumented with drift tubes and a muon tagger were used. During a 3-week period a dataset for analysis corresponding to $(3.27\pm 0.07)~\times ~10^{11}$ protons on target was recorded. This amounts to approximatively 1% of a SHiP spill.

The peptide SET-M33 is a molecule synthesized in tetra-branched form which is being developed as a new antibiotic against Gram-negative bacteria. Its isomeric form with D amino acids instead of the L version (SET-M33D) is also able to kill Gram-positive bacteria because of its higher resistance to bacterial proteases (Falciani et al., PLoS ONE, 2012, 7, e46259). Here we report the strong in vitro activity of SET-M33D (MIC range 0.7&ndash, 6.0 &micro, M) against multiresistant pathogens of clinical interest, including Gram-positives Staphylococcus aureus, Staphylococcus saprophyticus, and Enterococcus faecalis, and various Gram-negative enterobacteriaceae. SET-M33D antibacterial activity is also confirmed in vivo against a MRSA strain of S. aureus with doses perfectly compatible with clinical use (5 and 2.5 mg/Kg). Moreover, SET-M33D strongly neutralized lipopolysaccharide (LPS) and lipoteichoic acid (LTA), thus exerting a strong anti-inflammatory effect, reducing expression of cytokines, enzymes, and transcription factors (TNF-&alpha, IL6, COX-2, KC, MIP-1, IP10, iNOS, NF-&kappa, B) involved in the onset and evolution of the inflammatory process. These results, along with in vitro and in vivo toxicity data and the low frequency of resistance selection reported here, make SET-M33D a strong candidate for the development of a new broad spectrum antibiotic.

This article addresses the disparity in the transduction pathways for hypoxic and hypercapnic stimuli in carotid body glomus cells. We investigated and reviewed the experimental evidence showing that the response to hypoxia, but not to hypercapnia, is mediated by 1,4,5-inositol triphosphate receptors (IP3R/s) regulating the intracellular calcium content [Ca2+]c in glomus cells. The rationale was based on the past observations that inhibition of oxidative phosphorylation leads to the explicit inhibition of the hypoxic chemoreflex. [Ca2+]c changes were measured using cellular Ca2+-sensitive fluorescent probes, and carotid sinus nerve (CSN) sensory discharge was recorded with bipolar electrodes in in vitro perfused-superfused rat carotid body preparations. The cell-permeant, 2-amino-ethoxy-diphenyl-borate (2-APB; 100 μM) and curcumin (50 μM) were used as the inhibitors of IP3R/s. These agents suppressed the [Ca2+]c, and CSN discharge increases in hypoxia but not in hypercapnia, leading to the conclusion that only the hypoxic effects were mediated via modulation of IP3R/s. The ATP-induced Ca2+ release from intracellular stores in a Ca2+-free medium was blocked with 2-APB, supporting this conclusion.

Polycyclic aromatic hydrocarbon (PAH) exposure is widespread, and many PAHs are considered carcinogenic. The PAH-contaminated AWI Superfund site in Virginia provides a model for studying a complex PAH mixture and its extrapolation to cancer risk and PAH exposure in the general population. We examined cancer risk at the Superfund site due to sediment-derived PAHs and then evaluated PAH sources in the general population and potentially vulnerable subpopulations upon PAH mixture exposure. The PAH mixture was assessed for potential carcinogenicity using the US EPA’s OncoLogic™ ranking tool and the US EPA list of priority PAHs. Cancer risk due to PAH exposure was calculated for Superfund site users and compared to the US EPA assessment. Human intake and health endpoints of PAHs within the mixture were extracted from USEtox® chemical fate database, while mean intake exposure was calculated for U.S. adults for select PAHs using NHANES database urinary biomarkers. Eleven PAH compounds within the mixture were of carcinogenic concern, and seven PAHs conveyed significant excess cancer risk at the Superfund site and in the general population, wherein PAH-contaminated seafood ingestion was a main contributor. Other dietary sources of PAHs derived from PAH-contaminated soil or water could also play a role in total exposure. Vulnerable populations to PAH exposure and coinciding increased cancer risk may include, in addition to smokers, children and non-Hispanic blacks, which is a public health concern.

The ageing process is characterized by a decline in several physiological functions resulting in a reduced capability to maintain homeostasis. The lowered homeostatic capacity seems to involve the carotid body (CB), whose role is to modulate ventilation and tissue oxygen supply. It thus plays a prime role in all ageing processes. Ageing causes marked changes in CB morphology. In older animals, it is enlarged and shows a concomitant decrease in the percentage of chemoreceptor tissue, as well as a proliferation of type II cells. The carotid glomitis is present with aggregates of lymphocytes and fibrosis of the lobules. Type I cells are dehydrated, with a profound vacuolization, a shrinking nucleus, and lipofuscin accumulation. With increased age, human CB shows a reduction in the number and volume of mitochondria, fewer synaptic junctions between glomi, along with a reduction in CB content of neurotransmitters, leading to a sort of 'physiological denervation'. Ageing could be interpreted as a cumulative result of oxidative damage to cells, which derives from aerobic metabolism. Moreover, metabolic rate is tightly correlated with life duration; thus a loss in mitochondrial function is one of the prime factors affecting CB ageing processes. The age-related reduction in synaptic junctions might be a self-protective mechanism through which cells buffer themselves against an accumulation of reactive oxygen species. The correlation between hypoxia and the life duration of CB cells remains an open question until how and why cells sense oxygen is understood.

International audience; The paper aims to provide a new field-scale depositional model for the Upper Jurassic Arab Formation, in order to enlarge the stratigraphic and facies architecture knowledge in the onshore Abu Dhabi (UAE). The goal is reached following a threefold workflow structured by: (i) subsurface high-resolution facies analysis; (ii) well correlations; and (iii) paleofacies maps.The facies analysis was based on 540 m of cores and 277 thin sections. Eighteen facies were defined, and were grouped into five facies associations, representative of a shallow marine carbonate ramp succession. The ramp ranges from outer ramp to supratidal and intertidal environments, with a shoal complex protecting a lagoon. A micropaleontological approach helped at defining the field-scale evolution of the different sedimentary environments through time and space. The results were integrated with well log data (Gamma Ray and Density-Neutron Log), and were used to establish the stacking pattern of the Arab Formation. The vertical and lateral distribution of the facies associations in the area of study were highlighted by the well correlations. These latter were useful to generate several new paleofacies maps corresponding to key stratigraphic surfaces identified within the succession. The model reveals different directions of progradation, suggesting a local topographic control on the deposition of the Arab Formation. The cyclical arrangement of the facies associations and the peculiarities of this new model have been discussed at regional scale, suggesting a new paleogeographic scenario.

Curcumin, a phenolic compound extracted from Curcuma longa, exerts multiple pharmacological effects, including an antimicrobial action. Mycobacterium abscessus, an environmental, nontuberculous, rapidly growing mycobacterium, is an emerging human pathogen causing serious lung infections and one of the most difficult to treat, due to its multidrug resistance and biofilm-forming ability. We wanted to evaluate the antimicrobial and antivirulence activity of curcumin and its ability to synergize with antibiotics against a clinical M. abscessus strain (29904), isolated from the bronchoaspirate of a 66-year-old woman admitted to hospital for suspected tuberculosis. Curcumin [minimum inhibitory concentrations (MIC) = 128 mg/L] was synergic (fractional inhibitory concentration index ≤0.5) with amikacin, clarithromycin, ciprofloxacin, and linezolid, to which strain 29904 showed resistance/intermediate susceptibility. Curcumin at 1/8 × MIC significantly reduced motility, whereas at 4 × MIC, it completely inhibited 4- and 8-day mature biofilms. Synergistic combinations of curcumin and amikacin induced a general reduction in microbial aggregates and substantial loss in cell viability. Disruption of 4- and 8-day biofilms was the main effect detected when curcumin was the predominant compound. The present findings support previous evidence that curcumin is a potential antibiotic resistance breaker. Curcumin, either alone or combined with antibiotics, could provide a novel strategy to combat antibiotic resistance and virulence of M. abscessus.

Inouye et al. (2020) use the observation that Ser is coded in the genetic code by two blocks of codons that differ on more than one base to understand some aspects of the origin of the genetic code organization. I argue instead that this observation per se cannot be used to understand any aspect of the origin of the genetic code, unless it is accompanied by other assumptions concerning in the specific case: (i) the ancestrality of some amino acids, (ii) the hypothesis that the first mRNA to be translated was poly-G, which can be translated into poly-Gly, and (iii) an evolutionary mechanism for the genetic code origin based on the duplication of tRNAs. However, both the tRNA duplication mechanism and the existence of poly-G as the first mRNA to be translated are not corroborated as mechanisms through which the genetic code would have been structured. For example, the origin of the actual mRNA should have been preceded by the evolution of a proto-mRNA which evidently already coded for more than one amino acid. Therefore, when it evolved from proto-mRNA, the mRNA should already have coded for more than one amino acid. In other words, poly-G as mRNA would most likely never have existed because the first mRNAs already had to code for more than one amino acid. On the contrary, all these assumptions would have been operational if the observations of Inouye et al. (2020) had been discussed within the coevolution theory of the origin of the genetic code, which they do not.

Selenium (Se) may cause reproductive toxicity, yet the characteristics of Se bioaccumulation in aquatic food webs are understudied. Stream biofilms were grown in two reaches of Mud River, West Virginia (WV), including one downstream of a coal mine complex and an adjacent, unmined watershed. Mined stream biofilms contained significantly higher Se concentrations compared to unmined biofilms. An inverse relationship between water Se concentrations and biofilm accumulation factors was observed; mined-stream biofilms had an average bioconcentration factor (BCF) of 688 ± 350 fold while unmined-stream biofilms had an average BCF of 14505 ± 2700 fold.

The village of San Gregorio (SG), eight kilometres away from L'Aquila (central Italy), was severely damaged by the April 6, 2009 L'Aquila earthquake (M W 6.1). A coseismic fracture zone was mapped along SW-dipping fault segments crossing SG, which is situated at the base of a carbonate relief bounded by the Aterno river alluvial plain. An interdisciplinary approach was used to investigate the seismic response of the area based on geological-structural, geophysical and seismic analyses. We integrated our data with available information from the recent microzonation studies. SG is partly built on alluvial fan deposits constituted by cemented gravel, and partly on jointed carbonate bedrock. An extensive survey of noise measurements showed strong and polarized peaks in the horizontal-to-vertical spectral ratios (H/V), both on soft and rock sites in the 2–10 Hz frequency band. Further, we checked the stability with time of H/V ratios at three sites of SG. An analysis on local earthquakes confirmed the results of noise measurements. To understand the influence of rock mass jointing condition on site effects, we performed structural surveys on carbonate bedrock. We also evaluated the propagation velocities at rock sites using seismic active and seismic dilatometer test (SDMT) surveys. Our analysis showed low values of compressional (V P ) and shear wave (V S ) velocities of the outcropping rock, where we also observed strong H/V spectral peak and high-density rock fracturing.

Anthropogenic activities resulting in releases of selenium-laden waste streams threaten freshwater ecosystems. Lake ecosystems demand special consideration because they are characterized by prolonged retention of selenium and continuous cycling of the element through the food chain, through which it becomes available to toxicologically susceptible egg-laying vertebrates. This study documents the current selenium burden of lakes in North Carolina (NC) with historic selenium inputs from nearby coal-fired power plants. We measured selenium concentrations in surface waters, sediment pore waters, and resident fish species from coal combustion residual (CCR)-impacted lakes and paired reference lakes. The data are related to levels of recent selenium inputs and analyzed in the context of recently updated federal criteria for the protection of aquatic life. We show that the Se content of fish from lakes with the highest selenium inputs regularly exceed these criteria and are comparable to those measured during historic fish extirpation events in the United States. Large legacy depositions of CCRs within reservoir sediments are likely to sustain Se toxicity for many years despite recent laws to limit CCR discharge into surface waters in NC. Importantly, the widespread use of high-selenium coals for electricity generation extends the potential risk for aquatic ecosystem impacts beyond U.S. borders.

Several xenobiotic agents (e.g. metals, polycyclic aromatic hydrocarbons, nanoparticles, etc.) commonly involve the generation of reactive oxygen species (ROS) and oxidative stress as part of their toxic mode of action. Among piscine models, the zebrafish is a popular vertebrate model to study toxicity of various xenobiotic agents. Similarly to other vertebrates, zebrafish possess an extensive antioxidant system, including the reduced form of glutathione (GSH), which is an important antioxidant that acts alone or in conjunction with enzymes, such as glutathione peroxidase (GPx). Upon interaction with ROS, GSH is oxidized, resulting in the formation of glutathione disulfide (GSSG). GSSG is recycled by an auxiliary antioxidant enzyme glutathione reductase (GR). This article outlines detailed methods to measure the concentrations of GSH and GSSG, as well as the activities of GPx and GR in zebrafish larvae as robust and economical means to assess oxidative stress. The studies that have assessed these endpoints in zebrafish and alternative methods are also discussed. We conclude that the availability of these robust and economical methods support the use of zebrafish as a model organism in studies evaluating redox biology, as well as the induction of oxidative stress following exposure to toxic agents.

Myroides odoratimimus is an aerobic, non-fermenting Gram-negative multidrug-resistant bacterium widely distributed in nature that rarely causes infections in immunocompromised patients. We recently described in a diabetic patient a case of recurrent calcaneal ulcer infection caused by a M. odoratimimus strain showing potential for biofilm formation. For the first time, we therefore evaluated the ability of M. odoratimimus to form biofilm under different pH values and glucose concentrations using an in vitro “skin-like” model, and its susceptibility to levofloxacin, meropenem, and tigecycline. The expression of some antibiotic-resistance related genes was also monitored by RT-PCR during planktonic-to-biofilm transition. Our results indicated that M. odoratimimus can produce relevant amounts of biofilm biomass, in a time-dependent manner, especially at acidic pH and regardless of glucose concentration tested. The comparative analysis of MIC and MBC values between planktonic and sessile cells showed that resistance to antibiotics increased during the planktonic-to-biofilm transition. Viable cell count indicated that none of the tested antibiotics were able to completely eradicate preformed biofilms, although meropenem and levofloxacin were the most active causing a significant, dose-independent, reduction of biofilm's viability, as also confirmed by microscopic analysis. RT-PCR showed that antibiotic-resistance related gyrA and acrB genes are over-expressed during the transition from planktonic to sessile (biofilm) lifestyle. Overall, our findings showed that M. odoratimimus can form relevant amounts of inherently antibiotic-resistant biofilm under conditions relevant to wound site, therefore suggesting a role in the pathogenesis of chronic ulcer infections.

Common chronic conditions such as metabolic syndrome and diabetes are increasingly associated to metabolic and cardiovascular complications. Although Phyllanthus tenellus leaves have been used in decoctions as a popular remedy to control blood glucose levels and hypertension, its use needs a scientific basis. This study was therefore undertaken to report a phytochemical analysis of P. tenellus leaves and to test if the main active compound has potential to simultaneously tackle several pathophysiological features of metabolic syndrome and diabetes-related metabolic and vascular disorders such as hyperglycaemia, increased platelet activation, and endothelial dysfunction. We performed a partition of the methanolic extract of P. tenellus leaves among different organic solvents followed by chromatographic separation guided by the rat liver microsomal glucose-6-phosphatase assay. Two known tannins were identified by spectroscopic methods as pinocembrin-7-O-[3″-O-galloyl-4″,6″-(S)-hexahydroxydiphenoyl]-α-D-glucose, named P7OG by us, and gemin D. The structural determination of the isolated compounds was based on spectral data. The ability of the main active component, P7OG, to inhibit human platelet aggregation and to modify vascular reactivity of rat aortic rings incubated with high glucose (D-glucose 55 mM) was then evaluated. P7OG was further able to inhibit platelet aggregation induced by adenosine 5′-diphosphate and collagen, showed vasorelaxant effects in arteries precontracted with phenylephrine, and reverted the endothelium-dependent impairment effect of high glucose in rat aortic rings. In conclusion, one tannin isolated from P. tenellus showed promising metabolic, antiaggregant, and vascular effects, which suggests the potential beneficial use of P. tenellus to tackle complex cardiometabolic diseases.

Plastics are recognized as a worldwide threat to the environment, possibly affecting human health and wildlife. Small forms of plastics such as micro and nanoplastics can interact with other organic contaminants, potentially acting as chemical carriers and modulating their toxicity. In this study, we investigated the toxicity of polystyrene nanoparticles (Nano-PS) and a real-world environmental PAH mixture (Elizabeth River Sediment Extract – ERSE, comprised of 36 detected PAHs) to zebrafish embryos and larvae. Embryos were exposed to Nano-PS (0.1 – 10 ppm) or ERSE (0.1 – 5% v/v, equivalent to ΣPAH 5.07 – 25.36 ppb), or co-exposed to a combination of both. Larvae exposed to Nano-PS did not exhibit developmental defects, while larvae exposed to ERSE (2 – 5%) showed classic signs of PAH toxicity such as heart malformation and deformities in the jaw, fin, and tail. ERSE (5%) also impaired vascular development in the brain. When co-exposed, Nano-PS decreased the developmental deformities and impaired vascular development caused by ERSE. This was strongly correlated to the lower PAH bioaccumulation detected in the co-exposed animals (whole larvae, as well as the yolk sac, brain, and heart). Our data suggest that PAHs are sorbing to the surface of the Nano-PS, decreasing the concentration, uptake, and toxicity of free PAHs during the exposure. Such sorption of PAHs increases the agglomeration rate of Nano-PS during the exposure time, potentially decreasing the uptake of Nano-PS and associated PAHs. Despite that, similar induction of EROD activity was detected in animals exposed to ERSE in the presence or not of Nano-PS, suggesting that enough PAHs were accumulated in the organisms to induce cellular defense mechanisms. Nano-PS exposure (single or combined with ERSE) decreased the mitochondrial coupling efficiency and increased NADH production, suggesting and impairment on ATP production accompanied by a compensatory mechanism. Our data suggest that nanoplastics can sorb contaminants and potentially decrease their uptake due to particle agglomeration. Nanoplastics also target and disrupt mitochondrial energy production and act as vectors for the mitochondrial uptake of sorbed contaminants during embryonic and larval stages. Such negative effects of nanoplastics on energy metabolism and efficiency could be detrimental under multiple-stressors exposures and energy-demanding scenarios, what remains to be validated.

The most effective, safe and controlled way to perform tunnel excavations is through the use of Tunnel Boring Machines. This technology often requires the injection of chemicals to modify the soil properties, enhancing the control on the pressure applied at the front-face and consequently reducing the risks related to unexpected surface settlements, to excessive abrasion of the excavation tools and to clogging phenomena inside the working chamber. Since the reuse of the excavated soil is rapidly gaining importance, studies focused on the effect of these chemicals on soil properties, from both the geotechnical and environmental points of view, begin to be carried out more frequently in support of the design. This study addresses the evaluation of the effects of chemicals on the physical and mechanical properties of fine-grained soils: laboratory measurements of the effects on soil plasticity and on the associated variations in compressibility and undrained and residual strength of several samples of fine-grained soils from real tunnelling projects are presented. The results provide interesting insights on the interaction between the soil and the injected chemical, useful to better understand the most suitable use and dosage of the chemical and to consider its effects when reusing the conditioned soil.