Acute and Chronic Toxicity of Uncured Resin Feedstocks for Vat Photopolymerization 3D Printing to a Cladoceran (Ceriodaphnia Dubia).

The accessibility and popularity of additive manufacturing (AM) has increased over the past decade. Environmental hazard assessment and safety data sheets for 3D printer feedstocks has lagged technology development. Vat photopolymerization may have unique risks relative to other AM technologies due to mishandling of uncured monomers/oligomer feedstocks and its decreasing cost enabling uninformed residential use. The acute and chronic toxicity of six uncured resins to Ceriodaphnia dubia was explored. Two-day acute toxicity (LC50) ranged from 2.6 to 33 mg/L and inhibition concentrations (IC25) values for reproduction ranged from 0.33 to 16 mg/L. Cleaning and waste management procedures recommended in user guides could be the most hazardous handling scenario as use of isopropyl alcohol increases miscibility and thus the fate, transport and bioavailability of the uncured resins. Residential users may often be poorly informed about potential toxicity and the need for a plan for use, handling, and waste management of uncured resins.

Toward bioinspired polymer adhesives: activation assisted via HOBt for grafting of dopamine onto poly(acrylic acid).

The design of bioinspired polymers has long been an area of intense study, however, applications to the design of concrete admixtures for improved materials performance have been relatively unexplored. In this work, we functionalized poly(acrylic acid) (PAA), a simple analogue to polycarboxylate ether admixtures in concrete, with dopamine to form a catechol-bearing polymer (PAA-g-DA). Synthetic routes using hydroxybenzotriazole (HOBt) as an activating agent were examined for their ability in grafting dopamine to the PAA backbone. Previous literature using the traditional coupling reagent 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) to graft dopamine to PAA were found to be inconsistent and the sensitivity of EDC coupling reactions necessitated a search for an alternative. Additionally, HOBt allowed for greater control over per cent functionalization of the backbone, is a simple, robust reaction, and showed potential for scalability. This finding also represents a novel synthetic pathway for amide bond formation between dopamine and PAA. Finally, we performed preliminary adhesion studies of our polymer on rose granite specimens and demonstrated a 56% improvement in the mean adhesion strength over unfunctionalized PAA. These results demonstrate an early study on the potential of PAA-g-DA to be used for improving the bonds within concrete.

Green MIP-202(Zr) Catalyst: Degradation and Thermally Robust Biomimetic Sensing of Nerve Agents.

Rapid and robust sensing of nerve agent (NA) threats is necessary for real-time field detection to facilitate timely countermeasures. Unlike conventional phosphotriesterases employed for biocatalytic NA detection, this work describes the use of a new, green, thermally stable, and biocompatible zirconium metal-organic framework (Zr-MOF) catalyst, MIP-202(Zr). The biomimetic Zr-MOF-based catalytic NA recognition layer was coupled with a solid-contact fluoride ion-selective electrode (F-ISE) transducer, for potentiometric detection of diisopropylfluorophosphate (DFP), a F-containing G-type NA simulant. Catalytic DFP degradation by MIP-202(Zr) was evaluated and compared to the established UiO-66-NH2 catalyst. The efficient catalytic DFP degradation with MIP-202(Zr) at near-neutral pH was validated by 31P NMR and FT-IR spectroscopy and potentiometric F-ISE and pH-ISE measurements. Activation of MIP-202(Zr) using Soxhlet extraction improved the DFP conversion rate and afforded a 2.64-fold improvement in total percent conversion over UiO-66-NH2. The exceptional thermal and storage stability of the MIP-202/F-ISE sensor paves the way toward remote/wearable field detection of G-type NAs in real-world environments. Overall, the green, sustainable, highly scalable, and biocompatible nature of MIP-202(Zr) suggests the unexploited scope of such MOF catalysts for on-body sensing applications toward rapid on-site detection and detoxification of NA threats.

A Generalized Potentiostat Adaptor for Multiplexed Electroanalysis.

Electrochemical measurements over an array of electrodes may be accomplished with one of three potentiostat architectures: a single-channel device which averages the signal from a number of interconnected electrodes, a multichannel device with dedicated circuits for each electrode, or a single-channel device with a multiplexer interface to isolate the signal from specific electrodes. Of these three architectures, the use of a multiplexer interface is best suited to facilitate measurements over individual electrodes without the need for large numbers of dedicated potentiostat channels. We present a versatile strategy for the development of flexible printed circuit (FPC) electrode arrays with accompanying multiplexing hardware to interface with single-channel potentiostats. The FPC array was fabricated with 78 individually addressable 0.3 mm diameter gold working electrodes and characterized using optical and scanning electron microscopy, energy dispersive spectroscopy, profilometry, impedance spectroscopy, and cyclic voltammetry to investigate the morphology, elemental composition, height profile, impedance characteristics, and electrochemical response, respectively. Interfacing the FPC array via a simple connector with three 32-channel ADG731 multiplexers permitted electrochemical measurements using single-channel commercial potentiostats. Voltammetric experiments were conducted to demonstrate the reliability, stability, and reproducibility of the FPC array and interfacing hardware. The combination of these devices represents an accessible hardware platform with robust, functionalizable electrodes, a simple connection interface with commercial potentiostats, and a low cost through the use of off-the-shelf components. Our reported strategy holds great promise to facilitate multiplexed electroanalysis in next-generation sensors to increase statistical sample size and multianalyte detection capabilities.

In Situ Preconcentration and Quantification of Cu2+ via Chelating Polymer-Wrapped Multiwalled Carbon Nanotubes.

Trace analysis of heavy metals in complex, environmentally relevant matrices remains a significant challenge for electrochemical sensors employing stripping voltammetry-based detection schemes. We present an alternative method capable of selectively preconcentrating Cu2+ ions at the electrode surface using chelating polymer-wrapped multiwalled carbon nanotubes (MWCNTs). An electrochemical sensor consisting of poly-4-vinyl pyridine (P4VP)-wrapped MWCNTs anchored to a poly(ethylene terephthalate) (PET)-modified gold electrode (r = 1.5 mm) was designed, produced, and evaluated. The P4VP is shown to form a strong association with Cu2+ ions, permitting preconcentration adjacent to the electrode surface for interrogation via cyclic voltammetry. The sensor exhibited a detection limit of 0.5 ppm with a linear range of 1.1-13.8 ppm (16.6-216 µM) and a relative standard deviation (RSD) of 4.9% at the Environmental Protection Agency (EPA) limit of 1.3 ppm. Evaluation in tap water, lake water, ocean water, and deionized water rendered similar results, highlighting the generalizability of the presented preconcentration strategy. The advantages of electrochemical analysis paired with polymeric chelation represent an effective platform for the design and deployment of heavy metal sensors for continuous monitoring of natural waters.

Effect of Cellulose Nanofibrils and TEMPO-mediated Oxidized Cellulose Nanofibrils on the Physical and Mechanical Properties of Poly(vinylidene fluoride)/Cellulose Nanofibril Composites.

Cellulose nanofibrils (CNFs) are high aspect ratio, natural nanomaterials with high mechanical strength-to-weight ratio and promising reinforcing dopants in polymer nanocomposites. In this study, we used CNFs and oxidized CNFs (TOCNFs), prepared by a 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-mediated oxidation process, as reinforcing agents in poly(vinylidene fluoride) (PVDF). Using high-shear mixing and doctor blade casting, we prepared free-standing composite films loaded with up to 5 wt % cellulose nanofibrils. For our processing conditions, all CNF/PVDF and TOCNF/PVDF films remain in the same crystalline phase as neat PVDF. In the as-prepared composites, the addition of CNFs on average increases crystallinity, whereas TOCNFs reduces it. Further, addition of CNFs and TOCNFs influences properties such as surface wettability, as well as thermal and mechanical behaviors of the composites. When compared to neat PVDF, the thermal stability of the composites is reduced. With regards to bulk mechanical properties, addition of CNFs or TOCNFs, generally reduces the tensile properties of the composites. However, a small increase (~18%) in the tensile modulus was observed for the 1 wt % TOCNF/PVDF composite. Surface mechanical properties, obtained from nanoindentation, show that the composites have enhanced performance. For the 5 wt % CNF/PVDF composite, the reduced modulus and hardness increased by ~52% and ~22%, whereas for the 3 wt % TOCNF/PVDF sample, the increase was ~23% and ~25% respectively.

Silk-Like Protein with Persistent Radicals Identified in Oyster Adhesive by Solid-State NMR.

The cement produced by the Eastern oyster, Crassostrea virginica, may provide blueprints for waterproof biocompatible adhesives synthesized under benign conditions. The composition of this organic-inorganic composite, and of an organic extract, was characterized by 13C and 1H solid-state NMR and also compared with C. virginica shell and its organic extract. Quantification of the organic fraction by 13C and 1H NMR spectroscopy consistently showed 3 wt % organics in cement, which was higher than the 1.2 wt % in the shell. According to 13C NMR with spectral editing, the organic fraction of cement consisted of 73% protein, 25% polysaccharide, and 2% lipid. The organic acid-insoluble extract from the cement was mostly made up of protein remarkably rich in alanine and glycine. The unusual amino acid content matched the composition of silk-like proteins in the C. virginica or C. gigas genomes, including spidron-1-like and shelk2 previously found to be upregulated at the mantle edge. The corresponding extract from the shell contained 32% glycine and was also enriched in serine but not alanine, which was consistent with a previous wet-chemistry study. The 13C and 1H NMR spin-lattice relaxation in the organic component of cement and the acid-insoluble extract was 4-40 times faster than in the shell and showed pronounced nonexponentiality, indicating a high concentration of persistent radicals in the organic components of cement, in agreement with a prior EPR study. The presence of radicals in the acid-insoluble cement fraction was confirmed by observation of a paramagnetic shift anisotropy. 13C NMR corroborated prior observations that the calcium carbonate in the shell and pseudonacre was mostly calcite, whereas cement had an enhanced aragonite fraction. Surprisingly, 1H-13C NMR indicated that aragonite in cement was more distant from the organic fraction than was calcite. These results help advance our understanding of how oysters achieve adhesion within their wet environment.

Changes in Cementation of Reef Building Oysters Transitioning from Larvae to Adults.

Oysters construct extensive reef communities, providing food, protection from storms, and healthy coastlines. We still do not have a clear picture of how these animals attach to surfaces. Efforts described herein provide the first examination of adhesion at the transition from free swimming larvae to initial substrate attachment, through metamorphosis, and on to adulthood. Two different bonding systems were found to coexist. Larvae use an organic, hydrated glue that persists while the animal progresses into the juvenile phase, at which point a very different adhesive emerges. Juveniles bond with an organic-inorganic composite system, positioning the organic component for maximum adhesion by residing between the animal and substrate. Beyond understanding our marine environment, these insights may aid efforts in aquaculture, reef restoration, and adhesive design.

Genetically tunable M13 phage films utilizing evaporating droplets.

This effort utilizes a genetically tunable system of bacteriophage to evaluate the effect of charge, temperature and particle concentration on biomaterial synthesis utilizing the coffee ring (CR) effect. There was a 1.6-3 fold suppression of the CR at higher temperatures while maintaining self-assembled structures of thin films. This suppression was observed in phage with charged and uncharged surface chemistry, which formed ordered and disordered assemblies respectively, indicating CR suppression is not dependent on short-range ordering or surface chemistry. Analysis of the drying process suggests weakened capillary flow at elevated temperatures caused CR suppression and could be further enhanced for controlled assembly for advanced biomaterials.

Binding of Yeast Cytochrome c to Forty-Four Charge-Reversal Mutants of Yeast Cytochrome c Peroxidase: Isothermal Titration Calorimetry.

Previously, we constructed, expressed, and purified 46 charge-reversal mutants of yeast cytochrome c peroxidase (CcP) and determined their electronic absorption spectra, their reaction with H2O2, and their steady-state catalytic properties [ Pearl , N. M. et al. (2008) Biochemistry 47 , 2766 - 2775 ]. Forty-four of the mutants involve the conversion of either an aspartate or glutamate residue to a lysine residue, while two are positive-to-negative mutations, R31E and K149D. In this paper, we report on a calorimetric study of the interaction of each charge-reversal mutant (excluding the internal mutants D76K and D235K) with recombinant yeast iso-1 ferricytochrome c(C102T) (yCc) under conditions where only one-to-one yCc/CcP complex formation is observed. Thirteen of the 44 surface-site charge-reversal mutants decrease the binding affinity for yCc by a factor of 2 or more. Eight of the 13 mutations (E32K, D33K, D34K, E35K, E118K, E201K, E290K, E291K) occur within, or on the immediate periphery, of the crystallographically defined yCc binding site [ Pelletier , H. and Kraut , J. (1992) Science 258 , 1748 - 1755 ], three of the mutations (D37K, E98K, E209K) are slightly removed from the crystallographic site, and two of the mutations (D165K, D241K) occur on the "back-side" of CcP. The current study is consistent with a model for yCc binding to CcP in which yCc binds predominantly near the region defined by crystallographic structure of the 1:1 yCc-CcP complex, whether as a stable electron-transfer active complex or as part of a dynamic encounter complex.

Structural and compositional characterization of the adhesive produced by reef building oysters.

Oysters have an impressive ability to overcome difficulties of life within the stressful intertidal zone. These shellfish produce an adhesive for attaching to each other and building protective reef communities. With their reefs often exceeding kilometers in length, oysters play a major role in balancing the health of coastal marine ecosystems. Few details are available to describe oyster adhesive composition or structure. Here several characterization methods were applied to describe the nature of this material. Microscopy studies indicated that the glue is comprised of organic fiber-like and sheet-like structures surrounded by an inorganic matrix. Phospholipids, cross-linking chemistry, and conjugated organics were found to differentiate this adhesive from the shell. Symbiosis in material synthesis could also be present, with oysters incorporating bacterial polysaccharides into their adhesive. Oyster glue shows that an organic-inorganic composite material can provide adhesion, a property especially important when constructing a marine ecosystem.