A Classification Model to Identify Direct-Acting Mutagenic Polycyclic Aromatic Hydrocarbon Transformation Products.

Polycyclic aromatic hydrocarbons (PAHs) are a complex group of environmental contaminants, many having long environmental half-lives. As these compounds degrade, the changes in their structure can result in a substantial increase in mutagenicity compared to the parent compound. Over time, each individual PAH can potentially degrade into several thousand unique transformation products, creating a complex, constantly evolving set of intermediates. Microbial degradation is the primary mechanism of their transformation and ultimate removal from the environment, and this process can result in mutagenic activation similar to the metabolic activation that can occur in multicellular organisms. The diversity of the potential intermediate structures in PAH-contaminated environments renders hazard assessment difficult for both remediation professionals and regulators. A mixture of structural and energetic descriptors has proven effective in existing studies for classifying which PAH transformation products will be mutagenic. However, most existing studies of environmental PAH mutagens primarily focus on nitrogenated derivatives, which are prevalent in the atmosphere and not as relevant in soil. Additionally, PAH products commonly found in the environment can range from as large as five rings to as small as a single ring, requiring a broadly inclusive methodology to comprehensively evaluate mutagenic potential. We developed a combination of supervised and unsupervised machine learning methods to predict environmentally induced PAH mutagenicity with improved performance over currently available tools. K-means clustering with principal component analysis allows us to identify molecular clusters that we hypothesize to have similar mechanisms of action. Recursive feature elimination identifies the most influential descriptors. The cluster-specific regression outperforms available classifiers in predicting direct-acting mutagens resulting from the microbial biodegradation of PAHs and provides direction for future studies evaluating the environmental hazards resulting from PAH biodegradation.

Network Analysis for Prioritizing Biodegradation Metabolites of Polycyclic Aromatic Hydrocarbons.

Polycyclic aromatic hydrocarbons (PAHs) are a diverse group of environmental contaminants released during the combustion of organic materials and the production and utilization of fossil fuels. Once released, PAHs deposit in soil and water bodies where they are subjected to environmental transport and transformations. As they degrade, intermediate transformation products may play an important role in their environmental impact. However, studying the effects of these degradation products has proven challenging because of the complexity, transience, and low concentration of many intermediates. Herein, a novel integration of a pathway prediction system and network theory was developed and applied to a set of four PAHs to demonstrate a possible solution to this challenge. Network analysis techniques were employed to refine the thousands of potential outputs and elucidate compounds of interest. Using these tools, we determined correlations between PAH degradation network data and intermediate metabolite structures, gaining information about the chemical characteristics of compounds based on their placement within the degradation network. Upon applying our developed filtering algorithm, we are able to predict up to 48% of the most common transformation products identified in a comprehensive empirical literature review. Additionally, our integrated approach uncovers potential metabolites which connect those found by past empirical studies but are currently undetected, thereby filling in the gaps of information in PAH degradation pathways.

Structure-Property-Toxicity Relationships of Graphene Oxide: Role of Surface Chemistry on the Mechanisms of Interaction with Bacteria.

Graphene oxide (GO) is an antimicrobial agent with tunable surface chemistry. To identify the physicochemical determinants of GO's antimicrobial activity, we generated different modified Hummer's GO materials thermally annealed at 200, 500, or 800 °C (TGO200, TGO500, and TGO800, respectively) to modify the surface oxygen groups on the material. Plating assays show that as-received GO (ARGO) and TGO200, TGO500, and TGO800 reduce Escherichia coli viability by 50% (EC50) at 183, 143, 127, and 86 µg/mL, respectively, indicating higher bacterial toxicity as ARGO is reduced. To uncover the toxicity mechanism of GO, fluorescent dye-based assays were used to measure oxidative stress at the EC50. ARGO showed an increase in intracellular reactive oxygen species, measured as an increase in 2',7'-dichlorodihydrofluorescein diacetate fluorescence, whereas TGO500 and TGO800 induced an increase in the fluorescence of fluorescein diacetate (FDA) by 30 and 42%, suggesting a decrease in cell permeability. Because of a possible wrapping mechanism, plating assays after post-exposure sonication were performed to explain TGO's low oxidative response and high FDA levels. Results show no difference in colony-forming units, indicating that inhibition of cell growth is a result of the adsorption of bacterial cells on the GO material. By comparing different GO samples at their EC50, this study reveals that reduction of GO alters both the mechanisms of cellular interaction and the degree of toxicity to bacteria.

Life Cycle Impact and Benefit Trade-Offs of a Produced Water and Abandoned Mine Drainage Cotreatment Process.

A cotreatment process for produced water and abandoned mine drainage (AMD) has been established and demonstrated at the pilot-scale. The present study evaluates the potential of the proposed process to aid in management of two high volume wastewater resources in Pennsylvania. A systems-level approach is established to evaluate the primary trade-offs, including cotreatment process environmental impacts, transportation impacts, and environmental benefits realized from precluding direct AMD release to the environment. Life cycle impact assessment was used to quantify the environmental and human health impacts as well as to identify "hot spots" of the cotreatment process. Electricity use was found to be the dominant contributor to all impact categories. Extending the system boundary to include transportation of the two wastewaters to a to-be-determined cotreatment site revealed the important impact of transportation. An optimization approach was employed (using the region of Southwest Pennsylvania) to evaluate minimization of transportation distance considering the location and number of treatment sites. Finally, a quantitative analysis of environmental benefits realized by precluding direct AMD release to the environment was performed. The results suggest that the magnitude of benefit realized in treating a highly polluted AMD is greater than the magnitude of impacts from the cotreatment process.

Shape-Dependent Surface Reactivity and Antimicrobial Activity of Nano-Cupric Oxide.

Shape of engineered nanomaterials (ENMs) can be used as a design handle to achieve controlled manipulation of physicochemical properties. This tailored material property approach necessitates the establishment of relationships between specific ENM properties that result from such manipulations (e.g., surface area, reactivity, or charge) and the observed trend in behavior, from both a functional performance and hazard perspective. In this study, these structure-property-function (SPF) and structure-property-hazard (SPH) relationships are established for nano-cupric oxide (n-CuO) as a function of shape, including nanospheres and nanosheets. In addition to comparing these shapes at the nanoscale, bulk CuO is studied to compare across length scales. The results from comprehensive material characterization revealed correlations between CuO surface reactivity and bacterial toxicity with CuO nanosheets having the highest surface reactivity, electrochemical activity, and antimicrobial activity. While less active than the nanosheets, CuO nanoparticles (sphere-like shape) demonstrated enhanced reactivity compared to the bulk CuO. This is in agreement with previous studies investigating differences across length-scales. To elucidate the underlying mechanisms of action to further explain the shape-dependent behavior, kinetic models applied to the toxicity data. In addition to revealing different CuO material kinetics, trends in observed response cannot be explained by surface area alone. The compiled results contribute to further elucidate pathways toward controlled design of ENMs.

Designing nanomaterials to maximize performance and minimize undesirable implications guided by the Principles of Green Chemistry.

The Twelve Principles of Green Chemistry were first published in 1998 and provide a framework that has been adopted not only by chemists, but also by design practitioners and decision-makers (e.g., materials scientists and regulators). The development of the Principles was initially motivated by the need to address decades of unintended environmental pollution and human health impacts from the production and use of hazardous chemicals. Yet, for over a decade now, the Principles have been applied to the synthesis and production of engineered nanomaterials (ENMs) and the products they enable. While the combined efforts of the global scientific community have led to promising advances in the field of nanotechnology, there remain significant research gaps and the opportunity to leverage the potential global economic, societal and environmental benefits of ENMs safely and sustainably. As such, this tutorial review benchmarks the successes to date and identifies critical research gaps to be considered as future opportunities for the community to address. A sustainable material design framework is proposed that emphasizes the importance of establishing structure-property-function (SPF) and structure-property-hazard (SPH) relationships to guide the rational design of ENMs. The goal is to achieve or exceed the functional performance of current materials and the technologies they enable, while minimizing inherent hazard to avoid risk to human health and the environment at all stages of the life cycle.

Highly conductive single-walled carbon nanotube thin film preparation by direct alignment on substrates from water dispersions.

A safe, scalable method for producing highly conductive aligned films of single-walled carbon nanotubes (SWNTs) from water suspensions is presented. While microfluidic assembly of SWNTs has received significant attention, achieving desirable SWNT dispersion and morphology in fluids without an insulating surfactant or toxic superacid is challenging. We present a method that uniquely produces a noncorrosive ink that can be directly applied to a device in situ, which is different from previous fabrication techniques. Functionalized SWNTs (f-SWNTs) are dispersed in an aqueous urea solution to leverage binding between the amine group of urea and the carboxylic acid group of f-SWNTs and obtain urea-SWNT. Compared with SWNTs dispersed using conventional methods (e.g., superacid and surfactants), the dispersed urea-SWNT aggregates have a higher aspect ratio with a rodlike morphology as measured by light scattering. The Mayer rod technique is used to prepare urea-SWNT, highly aligned films (two-dimensional nematic order parameter of 0.6, 5 µm spot size, via polarized Raman) with resistance values as low as 15-1700 Ω/sq in a transmittance range of 2-80% at 550 nm. These values compete with the best literature values for conductivity of SWNT-enabled thin films. The findings offer promising opportunities for industrial applications relying on highly conductive thin SWNT films.

Life Cycle Payback Estimates of Nanosilver Enabled Textiles under Different Silver Loading, Release, And Laundering Scenarios Informed by Literature Review.

Silver was utilized throughout history to prevent the growth of bacteria in food and wounds. Recently, nanoscale silver has been applied to consumer textiles (nAg-textiles) to eliminate the prevalence of odor-causing bacteria. In turn, it is proposed that consumers will launder these items less frequently thus, reducing the life cycle impacts. While previous studies report that laundering processes are associated with the greatest environmental impacts of these textiles, there is no data available to support the proposed shift in consumer laundering behavior. Here, the results from a comprehensive literature review of nAg-textile life cycle studies are used to inform a cradle-to-grave life cycle impact assessment. Rather than assuming shifts in consumer behavior, the impact assessment is conducted in such a way that considers all laundering scenarios to elucidate the potential for reduced laundering to enable realization of a net life cycle benefit. In addition to identifying the most impactful stages of the life cycle across nine-midpoint categories, a payback period and uncertainty analysis quantifies the reduction in lifetime launderings required to recover the impacts associated with nanoenabling the textile. Reduction of nAg-textile life cycle impacts is not straightforward and depends on the impact category considered.

Life cycle impacts and benefits of a carbon nanotube-enabled chemical gas sensor.

As for any emerging technology, it is critical to assess potential life cycle impacts prior to widespread adoption to prevent future unintended consequences. The subject of this life cycle study is a carbon nanotube-enabled chemical gas sensor, which is a highly complex, low nanomaterial-concentration application with the potential to impart significant human health benefits upon implementation. Thus, the net lifecycle trade-offs are quantified using an impact-benefit ratio (IBR) approach proposed herein, where an IBR < 1 indicates that the downstream benefits outweigh the upstream impacts. The cradle-to-gate assessment results indicate that the midpoint impacts associated with producing CNTs are marginal compared with those associated with the other manufacturing stages. The cumulative upstream impacts are further aggregated to units of disability-adjusted life years (DALYs) using ReCiPe end point analysis method and quantitatively compared with the potential downstream DALY benefits, as lives saved, during the use phase. The approach presented in this study provides a guiding framework and quantitative method intended to encourage the development of nanoenabled products that have the potential to realize a net environmental, health, or societal benefit.

Toward tailored functional design of multi-walled carbon nanotubes (MWNTs): electrochemical and antimicrobial activity enhancement via oxidation and selective reduction.

Multiwalled carbon nanotubes (MWNTs) are utilized in a number of sectors as a result of their favorable electronic properties. In addition, MWNT antimicrobial properties can be exploited or considered a potential liability depending on their intended application and handling. The ability to tailor electrochemical and antimicrobial properties using economical and conventional treatment processes introduces the potential to significantly enhance product performance. Oxygen functional groups are known to influence several MWNT properties, including redox activity. Here, MWNTs were functionalized with oxygen groups using standard acid treatments followed by selective reduction via annealing. Chemical derivatization coupled to X-ray photoelectron spectroscopy was utilized to quantify specific surface oxygen group concentration after variable treatment conditions, which were then correlated to observed trends in electrochemical and antimicrobial activities. These activities were evaluated as the potential for MWNTs to participate in the oxygen reduction reaction and to have the ability to promote the oxidation of glutathione. The compiled results strongly suggest that the reduction of surface carboxyl groups and the redox activity of carbonyl groups promote enhanced MWNT reactivity and elucidate the opportunity to design functional MWNTs for enhanced performance in their intended electrochemical or antimicrobial application.

Particle-Driven Effects at the Bacteria Interface: A Nanosilver Investigation of Particle Shape and Dose Metric.

Design criteria for controlling engineered nanomaterial (ENM) antimicrobial performance will enable advances in medical, food production, processing and preservation, and water treatment applications. In pursuit of this goal, better resolution of how specific ENM properties, such as nanoparticle shape, influence antimicrobial activity is needed. This study probes the antimicrobial activity toward a model Gram-negative bacterium, Escherichia coli (E. coli), that results from interfacial interactions with differently shaped silver nanoparticles (AgNPs): cube-, disc-, and pseudospherical-AgNPs. The EC50 value (i.e., the concentration of AgNPs that inactivates 50% of the microbial population) for each shape is identified and presented as a function of mass, surface area, and particle number. Further, shifts in relative potency are identified from the associated dose-response curves (e.g., shifts left, to lower concentrations, indicate greater potency). When using a mass-based dose metric, the disc-AgNPs present the highest antimicrobial activity of the three shapes (EC50: 2.39 ± 0.26 µg/mL for discs, 2.99 ± 0.96 µg/mL for cubes, 116.33 ± 6.43 µg/mL for pseudospheres). When surface area and particle number are used as dose metrics, the cube-AgNPs possess the highest antimicrobial activity (EC50-surface area: 4.70 × 10-5 ± 1.51 × 10-5 m2/mL, EC50-particle: 5.97 × 109 ± 1.92 × 109 particles/mL), such that the relative trend in potency becomes cubes > discs > pseudospheres and cubes ≫ discs ⩾ pseudospheres, respectively. The results reveal that the antimicrobial potency of disc-AgNPs is sensitive to the dose metric, significantly decreasing in potency (∼5-30×) upon conversion from a mass-based concentration to surface area and particle number and influencing the conclusions drawn. The shift in relative particle potency highlights the importance of investigating various dose metrics within the experimental design and signals different particle parameters influencing shape-based antimicrobial activity. To probe shape-dependent behavior, we use a unique empirical approach where the physical and chemical properties (ligand chemistry, surface charge) of the AgNP shapes are carefully controlled, and total available surface area is equivalent across shapes as made through modifications to particle size and concentration. The results herein suggest that surface area alone does not drive antimicrobial activity as the different AgNP shapes at equivalent particle surface area yield significantly different magnitudes of antimicrobial activity (i.e., 100% inactivation for cube-AgNPs, <25% inactivation for disc- and pseudospherical-AgNPs). Further, the particle shapes studied possess different crystal facets, illuminating their potential influence on differentiating interactions between the particle surface and the microbe. Whereas surface area may partly contribute to antimicrobial activity in certain ENM shapes (i.e., disc-AgNPs in relation to the pseudospherical-AgNPs), the different magnitudes of antimicrobial activity across shape provide insight into the likely role of other particle-specific factors, such as crystal facets, driving the antimicrobial activity of other shapes (i.e., cube-AgNPs).

Similar toxicity mechanisms between graphene oxide and oxidized multi-walled carbon nanotubes in Microcystis aeruginosa.

In photosynthetic microorganisms, the toxicity of carbon nanomaterials (CNMs) is typically characterized by a decrease in growth, viability, photosynthesis, as well as the induction of oxidative stress. However, it is currently unclear how the shape of the carbon structure in CNMs, such as in the 1-dimensional carbon nanotubes (CNTs) compared to the two-dimensional graphene oxide (GO), affects the way they interact with cells. In this study, the effects of GO and oxidized multi-walled CNTs were compared in the cyanobacterium Microcystis aeruginosa to determine the similarities or differences in how the two CNMs interact with and induce toxicity to cyanobacteria. Using change in Chlorophyll a concentrations, the effective concentrations inducing 50% inhibition (EC50) at 96 h are found to be 11.1 µg/mL and 7.38 µg/mL for GO and CNTs, respectively. The EC50 of the two CNMs were not found to be statistically different. Changes in fluorescein diacetate and 2',7'-dichlorodihydrofluorescein diacetate fluorescence, measured at the EC50 concentrations, suggest a decrease in esterase enzyme activity but no oxidative stress. Scanning and transmission electron microscopy imaging did not show extensive membrane damage in cells exposed to GO or CNTs. Altogether, the decrease in metabolic activity and photosynthetic activity without oxidative stress or membrane damage support the hypothesis that both GO and CNTs induced indirect toxicity through physical mechanisms associated with light shading and cell aggregation. This indirect toxicity explains why the intrinsic differences in shape, size, and surface properties between CNTs and GO did not result in differences in how they induce toxicity to cyanobacteria.

Role of bacterial motility in differential resistance mechanisms of silver nanoparticles and silver ions.

Unlike conventional antimicrobials, the study of bacterial resistance to silver nanoparticles (AgNPs) remains in its infancy and the mechanism(s) through which it evolves are limited and inconclusive. The central question remains whether bacterial resistance is driven by the AgNPs, released Ag(I) ions or a combination of these and other factors. Here, we show a specific resistance in an Escherichia coli K-12 MG1655 strain to subinhibitory concentrations of AgNPs, and not Ag(I) ions, as indicated by a statistically significant greater-than-twofold increase in the minimum inhibitory concentration occurring after eight repeated passages that was maintained after the AgNPs were removed and reintroduced. Whole-population genome sequencing identified a cusS mutation associated with the heritable resistance that possibly increased silver ion efflux. Finally, we rule out the effect of particle aggregation on resistance and suggest that the mechanism of resistance may be enhanced or mediated by flagellum-based motility.

Unveiling the Synergistic Role of Oxygen Functional Groups in the Graphene-Mediated Oxidation of Glutathione.

This is the first report of an atomic-scale direct oxidation mechanism of the thiol group in glutathione (GSH) by epoxides on graphene oxide (GO) at room temperature. The proposed reaction mechanism is determined using a coupled experimental and computational approach; active sites for the reaction are determined through examination of GO surface chemistry changes before and after exposure to GSH, and density functional theory (DFT) calculations determine the reaction barriers for the possible GO-GSH reaction schemes. The findings build on the previously established catalytic mechanism of GSH oxidation by graphenic nanocarbon surfaces and importantly identify the direct reaction mechanism which becomes important in low-oxygen environments. Experimental results suggest epoxides as the active sites for the reaction with GSH, which we confirm using DFT calculations of reaction barriers and further identify a synergism between the adjacent epoxide and hydroxyl groups on the GO surface. The direct oxidation mechanism at specific oxygen sites offers insight into controlling GO chemical reactivity through surface chemistry manipulations. This insight is critical for furthering our understanding of GO oxidative stress pathways in cytotoxicity as well as for providing rational material design for GO applications that can leverage this reaction.

Guiding the design space for nanotechnology to advance sustainable crop production.

The globally recognized need to advance more sustainable agriculture and food systems has motivated the emergence of transdisciplinary solutions, which include methodologies that utilize the properties of materials at the nanoscale to address extensive and inefficient resource use. Despite the promising prospects of these nanoscale materials, the potential for large-scale applications directly to the environment and to crops necessitates precautionary measures to avoid unintended consequences. Further, the effects of using engineered nanomaterials (ENMs) in agricultural practices cascade throughout their life cycle and include effects from upstream-embodied resources and emissions from ENM production as well as their potential downstream environmental implications. Building on decades-long research in ENM synthesis, biological and environmental interactions, fate, transport and transformation, there is the opportunity to inform the sustainable design of nano-enabled agrochemicals. Here we perform a screening-level analysis that considers the system-wide benefits and costs for opportunities in which ENMs can advance the sustainability of crop-based agriculture. These include their on-farm use as (1) soil amendments to offset nitrogen fertilizer inputs, (2) seed coatings to increase germination rates and (3) foliar sprays to enhance yields. In each analysis, the nano-enabled alternatives are compared against the current practice on the basis of performance and embodied energy. In addition to identifying the ENM compositions and application approaches with the greatest potential to sustainably advance crop production, we present a holistic, prospective, systems-based approach that promotes emerging alternatives that have net performance and environmental benefits.

Opportunities and challenges for nanotechnology in the agri-tech revolution.

Current agricultural practices, developed during the green revolution, are becoming unsustainable, especially in the face of climate change and growing populations. Nanotechnology will be an important driver for the impending agri-tech revolution that promises a more sustainable, efficient and resilient agricultural system, while promoting food security. Here, we present the most promising new opportunities and approaches for the application of nanotechnology to improve the use efficiency of necessary inputs (light, water, soil) for crop agriculture, and for better managing biotic and abiotic stress. Potential development and implementation barriers are discussed, emphasizing the need for a systems approach to designing proposed nanotechnologies.

Copper release and transformation following natural weathering of nano-enabled pressure-treated lumber.

Commercially available lumber, pressure-treated with micronized copper azole (MCA), has largely replaced other inorganic biocides for residential wood treatment in the USA, yet little is known about how different outdoor environmental conditions impact the release of ionic, nano-scale, or larger (micron-scale) copper from this product. Therefore, we weathered pressure treated lumber for 18 months in five different climates across the continental United States. Copper release was quantified every month and local weather conditions were recorded continuously to determine the extent to which local climate regulated the release of copper from this nano-enabled product during its use phase. Two distinct release trends were observed: In cooler, wetter climates release occurred primarily during the first few months of weathering, as the result of copper leaching from surface/near-surface areas. In warmer, drier climates, less copper was initially released due to limited precipitation. However, as the wood dried and cracked, the exposed copper-bearing surface area increased, leading to increased copper release later in the product lifetime. Single-particle-ICP-MS results from laboratory prepared MCA-wood leachate solutions indicated that a) the predominant form of released copper passed through a filter smaller than 0.45 micrometers and b) released particles were largely resistant to dissolution over the course of 6 wks. Toxicity Characteristic Leaching Procedure (TCLP) testing was conducted on nonweathered and weathered MCA-wood samples to simulate landfill conditions during their end-of-life (EoL) phase and revealed that MCA wood released <10% of initially embedded copper. Findings from this study provide data necessary to complete a more comprehensive evaluation of the environmental and human health impacts introduced through release of copper from pressure treated lumber utilizing life cycle assessment (LCA).

A framework for sustainable nanomaterial selection and design based on performance, hazard, and economic considerations.

Engineered nanomaterials (ENMs) and ENM-enabled products have emerged as potentially high-performance replacements to conventional materials and chemicals. As such, there is an urgent need to incorporate environmental and human health objectives into ENM selection and design processes. Here, an adapted framework based on the Ashby material selection strategy is presented as an enhanced selection and design process, which includes functional performance as well as environmental and human health considerations. The utility of this framework is demonstrated through two case studies, the design and selection of antimicrobial substances and conductive polymers, including ENMs, ENM-enabled products and their alternatives. Further, these case studies consider both the comparative efficacy and impacts at two scales: (i) a broad scale, where chemical/material classes are readily compared for primary decision-making, and (ii) within a chemical/material class, where physicochemical properties are manipulated to tailor the desired performance and environmental impact profile. Development and implementation of this framework can inform decision-making for the implementation of ENMs to facilitate promising applications and prevent unintended consequences.

Toward safer multi-walled carbon nanotube design: Establishing a statistical model that relates surface charge and embryonic zebrafish mortality.

Given the increased utility and lack of consensus regarding carbon nanotube (CNT) environmental and human health hazards, there is a growing demand for guidelines that inform safer CNT design. In this study, the zebrafish (Danio rerio) model is utilized as a stable, sensitive biological system to evaluate the bioactivity of systematically modified and comprehensively characterized multi-walled carbon nanotubes (MWNTs). MWNTs were treated with strong acid to introduce oxygen functional groups, which were then systematically thermally reduced and removed using an inert temperature treatment. While 25 phenotypic endpoints were evaluated at 24 and 120 hours post-fertilization (hpf), high mortality at 24 hpf prevented further resolution of the mode of toxicity leading to mortality. Advanced multivariate statistical methods are employed to establish a model that identifies those MWNT physicochemical properties that best estimate the probability of observing an adverse outcome. The physicochemical properties considered in this study include surface charge, percent surface oxygen, dispersed aggregate size and morphology and electrochemical activity. Of the five physicochemical properties, surface charge, quantified as the point of zero charge (PZC), was determined as the best predictor of mortality at 24 hpf. From a design perspective, the identification of this property-hazard relationship establishes a foundation for the development of design guidelines for MWNTs with reduced hazard.