Active afforestation of drained peatlands is not a viable option under the EU Nature Restoration Law.

The EU Nature Restoration Law (NRL) is critical for the restoration of degraded ecosystems and active afforestation of degraded peatlands has been suggested as a restoration measure under the NRL. Here, we discuss the current state of scientific evidence on the climate mitigation effects of peatlands under forestry. Afforestation of drained peatlands without restoring their hydrology does not fully restore ecosystem functions. Evidence on long-term climate benefits is lacking and it is unclear whether CO2 sequestration of forest on drained peatland can offset the carbon loss from the peat over the long-term. While afforestation may offer short-term gains in certain cases, it compromises the sustainability of peatland carbon storage. Thus, active afforestation of drained peatlands is not a viable option for climate mitigation under the EU Nature Restoration Law and might even impede future rewetting/restoration efforts. Instead, restoring hydrological conditions through rewetting is crucial for effective peatland restoration.

The Effect of pH on the Viscoelastic Response of Alginate-Montmorillonite Nanocomposite Hydrogels.

Ionically cross-linked alginate hydrogels are used in a wide range of applications, such as drug delivery, tissue engineering, and food packaging. A shortcoming of these gels is that they lose their strength and degrade at low pH values. To develop gels able to preserve their integrity in a wide range of pH values, Ca-alginate-montmorillonite nanocomposite gels are prepared, and their chemical structure, morphology, and mechanical response are analyzed. As the uniformity of nanocomposite gels is strongly affected by concentrations of MMT and CaCl2, it is revealed that homogeneous gels can be prepared with 4 wt.% MMT and 0.5 M CaCl2 at the highest. The viscoelastic behavior of nanocomposite gels in aqueous solutions with pH = 7 and pH = 2 is investigated by means of small-amplitude compressive oscillatory tests. It is shown that Ca-alginate-MMT nanocomposite gels preserve their integrity while being swollen at pH = 2. The experimental data are fitted by a model with only two material parameters, which shows that the elastic moduli increase linearly with a concentration of MMT at all pH values under investigation due to formation of physical bonds between alginate chains and MMT platelets. The presence of these bonds is confirmed by ATR-FTIR spectroscopy. The morphology of nanocomposite gels is studied by means of wide-angle X-ray diffraction, which reveals that intercalation of polymer chains between clay platelets increases the interlayer gallery spacing.

Engineering Photo-Cross-Linkable MXene-Based Hydrogels: Durable Conductive Biomaterials for Electroactive Tissues and Interfaces.

Light-cured conductive hydrogels have attracted immense interest in the regeneration of electroactive tissues and bioelectronic interfaces. Despite the unique properties of MXene (MX), its light-blocking effect in the range of 300-600 nm hinders the efficient cross-linking of photocurable hydrogels. In this study, we investigated the photo-cross-linking process of MX-gelatin methacrylate (GelMa) composites with different types of photoinitiators and MX concentrations to prepare biocompatible, injectable, conductive, and photocurable composite hydrogels. The examined photoinitiators were Eosin Y, Irgacure 2959 (Type I), and lithium phenyl-2,4,6-trimethylbenzoyl phosphinate (Type II). The light-blocking effect of MX strongly affected the thickness, pore structure, swelling ratio, degradation, and mechanical properties of the light-cured hydrogels. Uniform distribution of MX in the hydrogel matrix was achieved at concentrations up to 0.04 wt % but the film thickness and curing times varied depending on the type of photoinitiator. It was feasible to prepare thin films (0.5 mm) by employing Type I photoinitiators under a relatively long light irradiation (4-5 min) while thick films with centimeter sizes could be rapidly cured by using Type II photoinitiator (<60 s). The mechanical properties, including elastic modulus, toughness, and stress to break for the Type II hydrogels were significantly superior (up to 300%) to those of Type I hydrogels depending on the MX concentration. The swelling ratio was also remarkably higher (648-1274%). A conductivity of about 1 mS/cm was attained at 0.1 mg/mL MX for the composite hydrogel cured by the Type I photoinitiator. In vitro cytocompatibility assays determined that the hydrogels promoted cell viability, metabolic activity, and robust proliferation of C2C12 myoblasts, which indicated their potential to support muscle cell growth during myogenesis. The developed photocurable GelMa-MX hydrogels have the potential to serve as bioactive and conductive scaffolds to modulate cellular functions and for tissue-device interfacing.

Methylotrophic Communities Associated with a Greenland Ice Sheet Methane Release Hotspot.

Subglacial environments provide conditions suitable for the microbial production of methane, an important greenhouse gas, which can be released from beneath the ice as a result of glacial melting. High gaseous methane emissions have recently been discovered at Russell Glacier, an outlet of the southwestern margin of the Greenland Ice Sheet, acting not only as a potential climate amplifier but also as a substrate for methane consuming microorganisms. Here, we describe the composition of the microbial assemblage exported in meltwater from the methane release hotspot at Russell Glacier and its changes over the melt season and as it travels downstream. We found that a substantial part (relative abundance 27.2% across the whole dataset) of the exported assemblage was made up of methylotrophs and that the relative abundance of methylotrophs increased as the melt season progressed, likely due to the seasonal development of the glacial drainage system. The methylotrophs were dominated by representatives of type I methanotrophs from the Gammaproteobacteria; however, their relative abundance decreased with increasing distance from the ice margin at the expense of type II methanotrophs and/or methylotrophs from the Alphaproteobacteria and Betaproteobacteria. Our results show that subglacial methane release hotspot sites can be colonized by microorganisms that can potentially reduce methane emissions.

Stable hydrogel adhesion to polydimethylsiloxane enables cyclic mechanical stimulation of 3D-bioprinted smooth muscle constructs.

During normal urination, smooth muscle cells (SMCs) in the lower urinary tract (LUT) are exposed to mechanical signals that have a critical impact on tissue structure and function. Nevertheless, the mechanisms underlying the maintenance of the contractile phenotype of SMCs remain poorly understood. This is due, in part, to a lack of studies that have examined the effects of mechanical loading using three-dimensional (3D) models. In this study, surface modifications of polydimethylsiloxane (PDMS) membrane were evaluated to investigate the effects of cyclic mechanical stimulation on SMC maturation in 3D constructs. Commercially available cell stretching plates were modified with amino or methacrylate groups to promote adhesion of 3D constructs fabricated by bioprinting. After 6 days of stimulation, the effects of mechanical stimulation on the expression of contractile markers at the mRNA and protein levels were analyzed. Methacrylate-modified surfaces supported stable adhesion of the 3D constructs to the membrane and facilitated cyclic mechanical stimulation, which significantly increased the expression of contractile markers at the mRNA and protein levels. These effects were found to be mediated by activation of the p38 MAPK pathway, as inhibition of this pathway abolished the effects of stimulation in a dose-dependent manner. These results provide valuable insights into the role of mechanical signaling in maintaining the contractile phenotype of bladder SMCs, which has important implications for the development of future treatments for LUT diseases.

Mechanical Properties of Alginate Hydrogels Cross-Linked with Multivalent Cations.

Ionically, cross-linked alginate gels have a potential to be used in a wide range of biomedical, environmental and catalytic applications. The study deals with preparation of alginate hydrogels cross-linked with various cations and the analysis of their equilibrium swelling and mechanical properties. It is shown that the type of cations used in the cross-linking process affects the elastic moduli and the equilibrium degree of swelling of the gels. The experimental data in small-amplitude oscillatory tests are fitted with a model that involves two material parameters: the elastic modulus of a polymer network and a measure of its inhomogeneity. The influence of cations on these quantities is studied numerically. It is revealed that the dependence of the elastic modulus of ionically cross-linked alginate gels on their equilibrium degree of swelling differs from that predicted by the conventional theory for covalently cross-linked gels.

The Effect of Temperature on the Mechanical Properties of Alginate Gels in Water/Alcohol Solutions.

Alginate organohydrogels prepared in water/alcohol mixtures play an important role in electronic and superconductor applications in low-temperature environments. The study deals with the preparation of Ca-alginate organohydrogels and the analysis of their equilibrium swelling and mechanical properties at sub-zero temperatures. It is shown that the equilibrium degree of swelling at room temperature is noticeably affected by the concentration of co-solvents (methanol, ethanol, and 2-propanol) in the mixtures and the number of carbon atoms in the co-solvent molecules. Mechanical properties are studied in small-amplitude oscillatory tests. The data are fitted with a model that involves three material parameters. The influence of temperature is investigated in temperature-sweep oscillatory tests under a cooling-heating program, where a noticeable difference is observed between the storage and loss moduli under cooling and heating (the hysteresis curves). The hysteresis areas are affected by the cooling/heating rate and the number of carbon atoms in the co-solvents.

Swelling of Homogeneous Alginate Gels with Multi-Stimuli Sensitivity.

A new two-step method is suggested for the preparation of homogeneous alginate gels. In the first step, alginate chains are weakly bonded by Ca2+ ions in an aqueous solution with a low pH. In the next step, the gel is immersed into a strong solution of CaCl2 to finalize the cross-linking process. Homogeneous alginate gels preserve their integrity in aqueous solutions with a pH ranging from 2 to 7 and ionic strength in the interval from 0 to 0.2 M, at temperatures ranging from room temperature up to 50 °C, and can be used in biomedical applications. The immersion of these gels into aqueous solutions with low pH induces the partial breakage of ionic bonds between chains (treated as gel degradation). This degradation affects the equilibrium and transient swelling of homogeneous alginate gels and makes them sensitive to the history of loading and environmental conditions (pH, ionic strength and temperature of aqueous solutions). As sensitivity to the environmental stimuli is a characteristic feature of polymer networks connected by catch bonds, homogeneous alginate gels may serve as a simple model, mimicking the behavior of more sophisticated structures in living matter.

Tailoring Hydrogel Composition and Stiffness to Control Smooth Muscle Cell Differentiation in Bioprinted Constructs.

BACKGROUND: Reliable in vitro cellular models are needed to study the phenotypic modulation of smooth muscle cells (SMCs) in health and disease. The aim of this study was to optimize gelatin methacrylate (GelMA)/alginate hydrogels for bioprinting three-dimensional (3D) SMC constructs. METHODS: Four different hydrogel groups were prepared by mixing different concentrations (% w/v) of GelMA and alginate: G1 (5/1.5), G2 (5/3), G3 (7.5/1.5), and G4 (7.5/3). GelMA 10% was used as control (G5). A circular structure containing human bladder SMCs was fabricated by using an extrusion-based bioprinter. The effects of the mixing ratios on printability, viability, proliferation, and differentiation of the cells were investigated. RESULTS: Rheological analysis showed that the addition of alginate significantly stabilized the change in mechanical properties with temperature variations. The group with the highest GelMA and alginate concentrations (G4) exhibited the highest viscosity, resulting in better stability of the 3D construct after crosslinking. Compared to other hydrogel compositions, cells in G4 maintained high viability (> 80%), exhibited spindle-shaped morphology, and showed a significantly higher proliferation rate within an 8-day period. More importantly, G4 provided an optimal environment for the induction of a SMC contractile phenotype, as evidenced by significant changes in the expression of marker proteins and morphological parameters. CONCLUSION: Adjusting the composition of GelMA/alginate hydrogels is an effective means of controlling the SMC phenotype. These hydrogels support bioprinting of 3D models to study phenotypic smooth muscle adaptation, with the prospect of using the constructs in the study of therapies for the treatment of urethral strictures.

Tuning the viscoelastic response of hydrogel scaffolds with covalent and dynamic bonds.

The processes of growth, proliferation and differentiation of stem cells encapsulated in 3D hydrogel microenvironments are strongly affected by the viscoelastic properties of the platforms. As the viscoelastic response of a hydrogel is determined by the rates of thermally induced dissociation of reversible cross-links, its modulation by introduction of several types of supramolecular and/or dynamic covalent bonds with different characteristic lifetimes has recently become a hot topic. To reduce the number of experiments needed for design of hydrogel microenvironments with required mechanical properties, a model is developed for the viscoelastic and viscoplastic responses of hydrogels with multiple networks bridged by covalent and physical bonds. An advantage of the model is that it (i) involves a small number of material parameters, (ii) describes observations in rheological and mechanical tests in a unified manner, and (iii) predicts conventional measures of viscoelasticity used in the analysis of viability of cells.

Thermo-Viscoelastic Response of Protein-Based Hydrogels.

Because of the bioactivity and biocompatibility of protein-based gels and the reversible nature of bonds between associating coiled coils, these materials demonstrate a wide spectrum of potential applications in targeted drug delivery, tissue engineering, and regenerative medicine. The kinetics of rearrangement (association and dissociation) of the physical bonds between chains has been traditionally studied in shear relaxation tests and small-amplitude oscillatory tests. A characteristic feature of recombinant protein gels is that chains in the polymer network are connected by temporary bonds between the coiled coil complexes and permanent cross-links between functional groups of amino acids. A simple model is developed for the linear viscoelastic behavior of protein-based gels. Its advantage is that, on the one hand, the model only involves five material parameters with transparent physical meaning and, on the other, it correctly reproduces experimental data in shear relaxation and oscillatory tests. The model is applied to study the effects of temperature, the concentration of proteins, and their structure on the viscoelastic response of hydrogels.

Equilibrium swelling of multi-stimuli-responsive copolymer gels.

Copolymer gels prepared by polymerization of thermo-responsive and anionic monomers demonstrate strong sensitivity to several triggers such as temperature, pH and ionic strength of aqueous solutions. For biomedical applications of these materials (as on-off switches in controlled drug delivery and release), fine tuning of their volume phase transition temperature (VPTT) and a sharp decay in degree of swelling upon transition from the swollen to the collapsed state are needed. These requirements are fulfilled under swelling of copolymer gels and microgels in water under acidic conditions, but are violated when tests are conducted under alkaline conditions or in aqueous solutions of salts with physiological salinity. A model is developed for equilibrium swelling of multi-stimuli-responsive copolymer gels in aqueous solutions with arbitrary pH and molar fractions of a monovalent salt. Unlike conventional approaches, the model accounts for secondary interactions between chains (hydrogen bonding) to describe the kinetics of aggregation of hydrophobic segments above VPTT. Material constants are determined by fitting experimental swelling diagrams on poly(N-isopropylacrylamide-co-sodium acrylate) gels with various molar fractions of ionic monomers. The effects of temperature, pH and molar fraction of salt on the equilibrium degree of swelling below and above VPTT are studied numerically.

Structure-property relations in linear viscoelasticity of supramolecular hydrogels.

Extraordinary mechanical properties of supramolecular gels (fracture toughness, fatigue resistance, injectability and self-healing ability) are strongly affected by their viscoelastic response driven by rearrangement (association and dissociation) of physical bonds. The kinetics of rearrangement is traditionally studied in small-amplitude shear oscillatory tests by analyzing the effect of the frequency of oscillations ω on the storage G' and loss G'' moduli. Conventional Maxwell-type models describe observations rather poorly when the gels reveal a pronounced flattening of the graphs G''(ω) at high frequencies. A simple model is derived in linear viscoelasticity of supramolecular gels. Its advantage is that the model reproduces experimental data correctly, on the one hand, and involves only four material constants, on the other. Based on the analysis of experimental data on gels cross-linked by coiled-coil complexes, covalent and ionic bonds, phenylboronic acid-diol complexes and metal-ligand coordination bonds, the model is applied to develop structure-property relations that describe the influence of chemical structure of supramolecular gels (concentration of polymer chains and type and molar fraction of temporary bonds) and environmental conditions (temperature, pH and ionic strength of buffer solutions) on their viscoelastic response.

Global Research Alliance N2 O chamber methodology guidelines: Design considerations.

Terrestrial ecosystems, both natural ecosystems and agroecosystems, generate greenhouse gases (GHGs). The chamber method is the most common method to quantify GHG fluxes from soil-plant systems and to better understand factors affecting their generation and mitigation. The objective of this study was to review and synthesize literature on chamber designs (non-flow-through, non-steady-state chamber) and associated factors that affect GHG nitrous oxide (N2 O) flux measurement when using chamber methods. Chamber design requires consideration of many facets that include materials, insulation, sealing, venting, depth of placement, and the need to maintain plant growth and activity. Final designs should be tailored, and bench tested, in order to meet the nuances of the experimental objectives and the ecosystem under study while reducing potential artifacts. Good insulation, to prevent temperature fluctuations and pressure changes, and a high-quality seal between base and chamber are essential. Elimination of pressure differentials between headspace and atmosphere through venting should be performed, and designs now exist to eliminate Venturi effects of earlier tube-type vent designs. The use of fans within the chamber headspace increases measurement precision but may alter the flux. To establish best practice recommendations when using fans, further data are required, particularly in systems containing tall plants, to systematically evaluate the effects that fan speed, position, and mixing rate have on soil gas flux.

Multiscale Characterization of a Wood-Based Biocrude as a Green Compatibilizing Agent for High-Impact Polystyrene/Halloysite Nanotube Nanocomposites.

This paper investigates merits of using a wood-based biocrude (WB) from aspen wood to improve the compatibility of halloysite nanotubes (HNTs) with high-impact polystyrene to develop nanocomposites with desirable thermomechanical properties. Morphological, thermal, and rheological properties of the resulting nanocomposite are used as indicators of the compatibility and dispersion of the modified HNT within the polymer matrix. Computational modeling using density functional theory is used along with laboratory experiments to provide a multiscale characterization of the above biocrude and nanocomposites. Studies performed through dispersion-corrected density functional theory calculations show that the active functional groups of WB molecules including carbonyl, hydroxyl, and carboxylic interact with the HNT surface, while their aromatic tails interact with the phenyl groups of the polystyrene. Furthermore, the studies reveal how WB molecules act as bridges between the hydrophobic polymer and the hydrophilic clay improving the compatibility. The latter was confirmed by Hansen solubility parameters and was evidenced in improved dispersion of clay within the polystyrene matrix observed by microscopy. Rheological and thermal analyses of the modified HNT and nanocomposites showed physical interactions of WB with HNT surface as well as interactions between the WB-modified HNT and the high-impact polystyrene. The WB was found to be a strong candidate as a green compatibilizing agent for HNT in high-impact polystyrene. The study results can provide insights for formulators and manufacturers looking for green compatibilizing agents in conventional nanocomposites for construction and manufacturing applications.

The potential role of miR-126, miR-21 and miR-10b as prognostic biomarkers in renal cell carcinoma.

Renal cell carcinoma (RCC) is the most commonly diagnosed renal tumor, consisting of ~3% of all malignancies worldwide. The prognosis of RCC can vary widely, and detecting patients at risk of recurrence at an early stage of disease may improve patient outcome. The factors presently used in a clinical setting cannot reliably predict the natural history of the disease. Therefore, there is a requirement to identify novel biomarkers that can aid in predicting patient outcome. Previous studies have indicated that microRNAs (miRNAs/miRs) are potential candidates as prognostic biomarkers for patients suffering from RCC. Consequently, the aims of the present study were to validate the potential of 3 of these miRNAs to predict the prognosis of patients with RCC, and to investigate the stability of endogenous control genes for miRNA studies in RCC tissues. The expression of 7 endogenous controls was measured using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in formalin-fixed paraffin-embedded tumor and benign tissues from patients suffering from clear cell RCC (ccRCC). The analyses identified RNU48 and U47 as the most stable endogenous controls. The expression of miR-126, miR-21 and miR-10b was analyzed using RT-qPCR in renal tissues from 116 patients diagnosed with ccRCC. All three investigated miRNAs were differentially expressed between malignant and benign tissues. miR-126 and miR-10b were also differentially expressed between grades and stages of ccRCC. In a univariate, but not in a multivariate model, low expression of miR-126 was associated with shorter time to recurrence of the disease. The results of the present study indicate that of the 3 miRNAs investigated, the expression of miR-126 has the strongest potential as a prognostic biomarker for patients suffering from ccRCC.

First observation of direct methane emission to the atmosphere from the subglacial domain of the Greenland Ice Sheet.

During a 2016 field expedition to the West Greenland Ice Sheet, a striking observation of significantly elevated CH4 concentrations of up to 15 times the background atmospheric concentration were measured directly in the air expelled with meltwater at a subglacial discharge point from the Greenland Ice Sheet. The range of hourly subglacial CH4 flux rate through the discharge point was estimated to be 3.1 to 134 g CH4 hr-1. These measurements are the first observations of direct emissions of CH4 from the subglacial environment under the Greenlandic Ice Sheet to the atmosphere and indicate a novel emission pathway of CH4 that is currently a non-quantified component of the Arctic CH4 budget.

To replicate, or not to replicate - that is the question: how to tackle nonlinear responses in ecological experiments.

A fundamental challenge in experimental ecology is to capture nonlinearities of ecological responses to interacting environmental drivers. Here, we demonstrate that gradient designs outperform replicated designs for detecting and quantifying nonlinear responses. We report the results of (1) multiple computer simulations and (2) two purpose-designed empirical experiments. The findings consistently revealed that unreplicated sampling at a maximum number of sampling locations maximised prediction success (i.e. the R² to the known truth) irrespective of the amount of stochasticity and the underlying response surfaces, including combinations of two linear, unimodal or saturating drivers. For the two empirical experiments, the same pattern was found, with gradient designs outperforming replicated designs in revealing the response surfaces of underlying drivers. Our findings suggest that a move to gradient designs in ecological experiments could be a major step towards unravelling underlying response patterns to continuous and interacting environmental drivers in a feasible and statistically powerful way.

Development and implementation of a near-real-time web reporting system on ground-level ozone in Europe.

This article presents the development and results of Ozone Web--near-real-time approach to communicate environmental information to policy makers, researchers, and the general public. In Ozone Web, ground-level ozone information from 750 air quality measurement stations across Europe is collected on an hourly basis, filtered, interpolated, and presented on zoomable maps and graphs. Compared with other environmental information initiatives, the main aspects of this Website is the allowance for user interactivity, free access to data, and high timeliness. Data are published 2 to 3 h after actual monitoring. In a response to the acute characteristics of air pollution, the basic principle is that up-to-date and accurate information about air pollution levels will help (1) citizens to protect their health, (2) policy makers in assessing the state of the environment, and (3) researchers in exchanging data and knowledge. Near-real-time information systems on the Web seem to be a valuable complement to future environmental reporting, and the European Environment Agency is currently investigating the requirements needed to extend the use of near-real-time data, including reporting on air pollutants other than ozone.

The microbial ecology of processing equipment in different fish industries-analysis of the microflora during processing and following cleaning and disinfection.

The microflora adhering to the processing equipment during production and after cleaning and disinfecting procedures was identified in four different processing plants. A total of 1009 microorganisms was isolated from various-agar plates and identified. A stepwise procedure using simple phenotypic tests was used to identify the isolates and proved a fast way to group a large collection of microorganisms. Pseudomonas, Neisseriaceae, Enterobactericeae, Coryneform, Acinetobacter and lactic acid bacteria dominated the microflora of cold-smoked salmon plants, whereas the microflora in a plant processing semi-preserved herring consisted of Pseudomonas, Alcaligenes and Enterobactericeae. Psychrobacter, Staphylococcus and yeasts were found in a caviar processing plant. Overall, many microorganisms that are often isolated from fish were also isolated from the fish processing plants. However, some selection depending on processing parameters occurred, since halo- and osmo-tolerant organisms dominated in the caviar processing. After cleaning and disinfection, yeasts, Pseudomonas, Neisseriaceae and Alcaligenes remained in smokehouses, yeasts and Pseudomonas in the herring plant and Pseudomonas, Staphylococcus and yeasts in the caviar plant. The dominant adhering organisms after cleaning and disinfection were pseudomonads and yeasts independently of the microflora during processing. Knowledge of the adhering microflora is essential in the Good Hygienic Practises programme of food processing plants, as the development and design of improved cleaning and disinfecting procedures should target the microorganisms persisting and potentially contaminating the product.