The Impact of Activating Agents on Non-Enzymatic Nucleic Acid Extension Reactions.

Non-enzymatic template-directed primer extension is increasingly being studied for the production of RNA and DNA. These reactions benefit from producing RNA or DNA in an aqueous, protecting group free system, without the need for expensive enzymes. However, these primer extension reactions suffer from a lack of fidelity, low reaction rates, low overall yields, and short primer extension lengths. This review outlines a detailed mechanistic pathway for non-enzymatic template-directed primer extension and presents a review of the thermodynamic driving forces involved in entropic templating. Through the lens of entropic templating, the rate and fidelity of a reaction are shown to be intrinsically linked to the reactivity of the activating agent used. Thus, a strategy is discussed for the optimization of non-enzymatic template-directed primer extension, providing a path towards cost-effective in vitro synthesis of RNA and DNA.

Electrospinning Triboelectric Laminates: A Pathway for Scaling Energy Harvesters.

Herein, a new paradigm of triboelectric polymers-the triboelectric laminate-a volumetric material with electromechanical response comparable to the benchmark soft piezoelectric material polyvinylidene difluoride is reported. The electromechanical response in the triboelectric laminate arises from aligned dipoles, generated from the orientation of contact electrification in the laminates bulk volume. The dipoles form between sequential bilayers consisting of two different electrospun polymer fibers of different diameter. The loose interface between the fiber bilayers ensures friction and triboelectric charging between two polymers. The electric output from the electrospun triboelectric laminate increases with increasing density of the bilayers. This system design has clear benefits over other flexible devices for mechanical energy harvesting as it does not require any poling procedures, and the electromechanical response is stable over 24 h of continuous operation. Moreover, the electromechanically responsive electrospun laminate can be made from all types of polymers, thus providing ample room for further improvements or functionalities such as stretchability, biodegradability, or biocompatibility. The concept of a triboelectric laminate can be introduced into existing triboelectric nanogenerator form factors, to dramatically increase charge harvesting of a variety of devices.

Biofunctionality with a twist: the importance of molecular organisation, handedness and configuration in synthetic biomaterial design.

The building blocks of life - nucleotides, amino acids and saccharides - give rise to a large variety of components and make up the hierarchical structures found in Nature. Driven by chirality and non-covalent interactions, helical and highly organised structures are formed and the way in which they fold correlates with specific recognition and hence function. A great amount of effort is being put into mimicking these highly specialised biosystems as biomaterials for biomedical applications, ranging from drug discovery to regenerative medicine. However, as well as lacking the complexity found in Nature, their bio-activity is sometimes low and hierarchical ordering is missing or underdeveloped. Moreover, small differences in folding in natural biomolecules (e.g., caused by mutations) can have a catastrophic effect on the function they perform. In order to develop biomaterials that are more efficient in interacting with biomolecules, such as proteins, DNA and cells, we speculate that incorporating order and handedness into biomaterial design is necessary. In this review, we first focus on order and handedness found in Nature in peptides, nucleotides and saccharides, followed by selected examples of synthetic biomimetic systems based on these components that aim to capture some aspects of these ordered features. Computational simulations are very helpful in predicting atomic orientation and molecular organisation, and can provide invaluable information on how to further improve on biomaterial designs. In the last part of the review, a critical perspective is provided along with considerations that can be implemented in next-generation biomaterial designs.

A bright future for engineering piezoelectric 2D crystals.

The piezoelectric effect, mechanical-to-electrical and electrical-to-mechanical energy conversion, is highly beneficial for functional and responsive electronic devices. To fully exploit this property, miniaturization of piezoelectric materials is the subject of intense research. Indeed, select atomically thin 2D materials strongly exhibit the piezoelectric effect. The family of 2D crystals consists of over 7000 chemically distinct members that can be further manipulated in terms of strain, functionalization, elemental substitution (i.e. Janus 2D crystals), and defect engineering to induce a piezoelectric response. Additionally, most 2D crystals can stack with other similar or dissimilar 2D crystals to form a much greater number of complex 2D heterostructures whose properties are quite different to those of the individual constituents. The unprecedented flexibility in tailoring 2D crystal properties, coupled with their minimal thickness, make these emerging highly attractive for advanced piezoelectric applications that include pressure sensing, piezocatalysis, piezotronics, and energy harvesting. This review summarizes literature on piezoelectricity, particularly out-of-plane piezoelectricity, in the vast family of 2D materials as well as their heterostructures. It also describes methods to induce, enhance, and control the piezoelectric properties. The volume of data and role of machine learning in predicting piezoelectricity is discussed in detail, and a prospective outlook on the 2D piezoelectric field is provided.

Laying Waste to Mercury: Inexpensive Sorbents Made from Sulfur and Recycled Cooking Oils.

Mercury pollution threatens the environment and human health across the globe. This neurotoxic substance is encountered in artisanal gold mining, coal combustion, oil and gas refining, waste incineration, chloralkali plant operation, metallurgy, and areas of agriculture in which mercury-rich fungicides are used. Thousands of tonnes of mercury are emitted annually through these activities. With the Minamata Convention on Mercury entering force this year, increasing regulation of mercury pollution is imminent. It is therefore critical to provide inexpensive and scalable mercury sorbents. The research herein addresses this need by introducing low-cost mercury sorbents made solely from sulfur and unsaturated cooking oils. A porous version of the polymer was prepared by simply synthesising the polymer in the presence of a sodium chloride porogen. The resulting material is a rubber that captures liquid mercury metal, mercury vapour, inorganic mercury bound to organic matter, and highly toxic alkylmercury compounds. Mercury removal from air, water and soil was demonstrated. Because sulfur is a by-product of petroleum refining and spent cooking oils from the food industry are suitable starting materials, these mercury-capturing polymers can be synthesised entirely from waste and supplied on multi-kilogram scales. This study is therefore an advance in waste valorisation and environmental chemistry.

A DNA Circuit for IsomiR Detection.

A synthetic DNA oligonucleotide has been programmed to function as a biological circuit to detect 5'-IsomiRs. The circuit consists of two integrated DNA switches. The first is "activated" when a DNA probe is enzymatically modified by a reverse transcriptase that incorporates nucleotides complementary to the 5'-region of a microRNA (miRNA). Addition of the correct number and sequence of nucleotides enables the probe to assemble into an asymmetric DNA hairpin. The reconfigured hairpin probe then primes an internal polymerisation reaction, resulting in the synthesis of a symmetrical DNA hairpin. This activates the second switch, which then initiates the amplification of reverse-transcribed miRNA through a continuous cycle of DNA nicking and polymerisation. The DNA circuit enables sensitive and rapid detection of femtomoles of a miRNA transcript under isothermal conditions.

Approaches for the detection of harmful algal blooms using oligonucleotide interactions.

Blooms of microscopic algae in our waterways are becoming an increasingly important environmental concern. Many are sources of harmful biotoxins that can lead to death in humans, marine life and birds. Additionally, their biomass can cause damage to ecosystems such as oxygen depletion, displacement of species and habitat alteration. Globally, the number and frequency of harmful algal blooms has increased over the last few decades, and monitoring and detection strategies have become essential for managing these events. This review discusses developments in the use of oligonucleotide-based 'molecular probes' for the selective monitoring of algal cell numbers. Specifically, hybridisation techniques will be a focus.

Planar silver nanowire, carbon nanotube and PEDOT:PSS nanocomposite transparent electrodes.

Highly conductive, transparent and flexible planar electrodes were fabricated using interwoven silver nanowires and single-walled carbon nanotubes (AgNW:SWCNT) in a PEDOT:PSS matrix via an epoxy transfer method from a silicon template. The planar electrodes achieved a sheet resistance of 6.6 ± 0.0 Ω/□ and an average transmission of 86% between 400 and 800 nm. A high figure of merit of 367 Ω-1 is reported for the electrodes, which is much higher than that measured for indium tin oxide and reported for other AgNW composites. The AgNW:SWCNT:PEDOT:PSS electrode was used to fabricate low temperature (annealing free) devices demonstrating their potential to function with a range of organic semiconducting polymer:fullerene bulk heterojunction blend systems.

Sequence selective capture, release and analysis of DNA using a magnetic microbead-assisted toehold-mediated DNA strand displacement reaction.

This paper reports on the modification of magnetic beads with oligonucleotide capture probes with a specially designed pendant toehold (overhang) aimed specifically to capture double-stranded PCR products. After capture, the PCR products were selectively released from the magnetic beads by means of a toehold-mediated strand displacement reaction using short artificial oligonucleotide triggers and analysed using capillary electrophoresis. The approach was successfully shown on two genes widely used in human DNA genotyping, namely human c-fms (macrophage colony-stimulating factor) proto-oncogene for the CSF-1 receptor (CSF1PO) and amelogenin.

Understanding and Controlling Electrostatic Discharge in Triboelectric Nanogenerators.

Triboelectric nanogenerators (TENGs) have been widely used to harness various forms of mechanical energy for conversion to electrical energy. However, the contentious challenge in characterising TENGs is the lack of standard protocols for assessing mechanical-to-electrical energy conversion processes. Herein, macroscopic signal analysis is used to identify three key charging events within triboelectric signals: charge induction (CI), contact electrification (CE), and electrostatic discharge (ESD). By considering two phases of motion during contact-separation (approach and departure of the contact materials), CI arising from the motion of bound surface charge (varying electric field) between opposing contact materials is shown to dominate the measured displacement current signal, rather than the process of CE itself. Furthermore, the conventional signal (i. e., voltage, current, charge) interpretation of CE and CI during approach and departure phases is re-assessed, to indicate that the sudden spike of current often observed immediately prior to contact (or after separation) arises from polarity inverting electrostatic discharge (ESD). This aspect of the measured triboelectric effect, which is often ignored, is crucial for the design of TENGs and hence, techniques to enhance the understanding and control over the stochastic occurrence of ESDs is explored. The methods proposed for the deconvolution of the macroscopic signal components of TENGs, and mitigation of ESD occurrences, will allow for precise quantification of the associated charging events. The applications of this study will template the design and development of future super-TENGs with optimised energy conversion capabilities.

Toehold-mediated nonenzymatic DNA strand displacement as a platform for DNA genotyping.

Toehold-mediated DNA strand displacement provides unique advantages in the construction and manipulation of multidimensional DNA nanostructures as well as nucleic acid sequence analysis. We demonstrate a step change in the use of toehold-mediated DNA strand displacement reactions, where a double-stranded DNA duplex, containing a single-stranded toehold domain, enzymatically generated and then treated as a molecular target for analysis. The approach was successfully implemented for human DNA genotyping, such as gender identification where the amelogenin gene was used as a model target system, and detecting single nucleotide polymorphisms of human mitochondrial DNA. Kinetics of the strand displacement was monitored by the quenched Förster resonance energy transfer effect.

On-chip capacitively coupled contactless conductivity detection using "injected" metal electrodes.

We demonstrate the use of injected gallium electrodes for capacitively coupled contactless conductivity detection (C(4)D) within a microchip electrophoresis device. Evaluation of the electrodes for quantitative detection of electrophoretically separated lithium, sodium and potassium ions showed the system offers competitive detection limits of 6.1 × 10(-6) M, 6.7 × 10(-6) M and 8.5 × 10(-6) M, respectively. The fabrication process is fast, highly reproducible, and eliminates difficulties with electrode alignment. Using this approach C(4)D can be readily achieved in any microchip by simply adding extra 'electrode' channels to the microchip design.

Rapid separation of synthetic oligonucleotides on polymer modified capillary surfaces using short-end injection capillary electrophoresis in free solution.

Here we use short-end electrokinetic injection capillary electrophoresis (CE) to investigate the free solution mobility of short strands of double-stranded oligonucleotides (dsODNs) on polymer modified capillaries. Single base pair (bp) resolution (Rs) of dsODNs ranging from 16-20 bp was achieved in free solution on an 8 cm capillary dynamically coated with poly(ethylpyrrolidine methacrylate-co-methyl methacrylate) (PEPyM-co-PMMA) random copolymer. Interestingly, separation of a dsODN mixture containing two 16 bp strands of different sequences resulted in partial resolution (0.52) implying that the free solution mobility of dsODNs was sequence dependent. The single bp resolution achieved for the complementary sequence strands (the sequence of all strands in the mixtures contained the same 16 bp sequence) was improved by up to 37% for separation of dsODNs containing non-complementary sequences. The 16 bp peak was not additive within each mixture, indicating the presence of ODN-ODN interactions. Investigation of these interactions (and ODN-buffer interactions) showed that they can be influenced by the ionic strength and conductivity of the background electrolyte (BGE). Increasing the ionic strength reduced the ODN-ODN interactions and improved the resolution, whereas, increasing the conductivity reduced ODN-buffer interactions, increasing the mobility, at the consequence of promoting ODN-ODN interactions, and hence decreasing the resolution.

29Si{1H} CP-MAS NMR comparison and ATR-FTIR spectroscopic analysis of the diatoms Chaetoceros muelleri and Thalassiosira pseudonana grown at different salinities.

Diatoms are key indicators of marine environmental health. To further understand how diatoms respond to varying degrees of salinity, either due to climate change or brine waste discharge into marine environments, two different diatom species were studied. Thalassiosira pseudonana and Chaetoceros muelleri were cultured at three different salinities namely, 26 practical salinity units (PSU or parts per thousand), 36 PSU (standard salinity for culturing of seawater species) and 46 PSU. Changes in silica and organic content within the cultured diatoms were analysed using solid-state (29)Si{(1)H} cross-polarization-magic angle spinning (CP-MAS) nuclear magnetic resonance (NMR) and attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopies coupled with analysis of variance. (29)Si CP-MAS NMR showed that qualitatively the Q4:Q3 area ratios of C. muelleri, grown away from standard salinities, increased in response to the formation of more condensed (2 ≡SiOH → ≡Si-O-Si≡ + H2O) and/or an increase in closely associated organic matter to the Q4 component of the diatoms. This was not observed for T. pseudonana. However, both species showed the appearance of a new peak centered at 1575-1580 cm(-1) in the ATR-FTIR spectra, designated as the C═N band of nitrogenous purine-type compounds. Further, the C. muelleri species was shown to produce more extracellular polymeric substances at non-standard salinities. On this basis, results suggest that there is a strong relationship between diatom composition and salinity and that C. muelleri is more sensitive to its environment than T. pseudonana.

Highly conductive interwoven carbon nanotube and silver nanowire transparent electrodes.

Electrodes fabricated using commercially available silver nanowires (AgNWs) and single walled carbon nanotubes (SWCNTs) produced sheet resistances in the range 4-24 Ω â–¡-1 with specular transparencies up to 82 %. Increasing the aqueous dispersibility of SWCNTs decreased the bundle size present in the film resulting in improved SWCNT surface dispersion in the films without compromising transparency or sheet resistance. In addition to providing conduction pathways between the AgNW network, the SWCNTs also provide structural support, creating stable self-supporting films. Entanglement of the AgNWs and SWCNTs was demonstrated to occur in solution prior to deposition by monitoring the transverse plasmon resonance mode of the AgNWs during processing. The interwoven AgNW/SWCNT structures show potential for use in optoelectronic applications as transparent electrodes and as an ITO replacement.

Evaluation of a lanthanide nanoparticle-based contrast agent for microcomputed tomography of porous channels in subchondral bone.

Osteoarthritis (OA) is a chronic joint disease that causes disability and pain. The osteochondral interface is a gradient tissue region that plays a significant role in maintaining joint health. It has been shown that during OA, increased neoangiogenesis creates porous channels at the osteochondral interface allowing the transport of molecules related to OA. Importantly, the connection between these porous channels and the early stages of OA development is still not fully understood. Microcomputed tomography (microCT) offers the ability to image the porous channels at the osteochondral interface, however, a contrast agent is necessary to delineate the different X-ray attenuations of the tissues. In this study BaYbF5 -SiO2 nanoparticles are synthesized and optimized as a microCT contrast agent to obtain an appropriate contrast attenuation for subsequent segmentation of structures of interest, that is, porous channels, and mouse subchondral bone. For this purpose, BaYbF5 nanoparticles were synthesized and coated with a biocompatible silica shell (SiO2 ). The optimized BaYbF5 -SiO2 27 nm nanoparticles exhibited the highest average microCT attenuation among the biocompatible nanoparticles tested. The BaYbF5 -SiO2 27 nm nanoparticles increased the mean X-ray attenuation of structures of interest, for example, porous channel models and mouse subchondral bone. The BaYbF5 -SiO2 contrast attenuation was steady after diffusion into mouse subchondral bone. In this study, we obtained for the first time, the average microCT attenuation of the BaYbF5 -SiO2 nanoparticles into porous channel models and mouse subchondral bone. In conclusion, BaYbF5 -SiO2 nanoparticles are a potential contrast agent for imaging porous channels at the osteochondral interface using microCT.

Quantifying environmental emissions of microplastics from urban rivers in Melbourne, Australia.

This study aims to understand the amount and type of microplastics flowing into Port Phillip Bay from urban rivers around Melbourne. Water samples were collected from the Patterson, Werribee, Maribyrnong, and Yarra Rivers, which contribute 97 % to the total flow into Port Phillip Bay. On average, the rivers contained a mean of 9 ± 15 microplastics/L and ranged from 4 ± 3 microplastics/L (Patterson) to 22 ± 11 microplastics/L (Werribee). Of the eight polymers investigated, polyamide and polypropylene were the most frequently detected polymers. Using the mean concentration of each river, the flow of microplastics into Port Philip Bay was estimated to be 7.5 × 106 microplastics per day and 3.7 × 1010 microplastics per year. To fully understand the fate and transport of microplastics into Port Phillip Bay, this study would be the foundation for a more in-depth investigation. Here, further samples will be collected at more points along the river and at the midpoint of each season.

Macro-Size Regenerated Cellulose Fibre Embedded with Graphene Oxide with Antibacterial Properties.

Macro-size regenerated cellulose fibres (RCFs) with embedded graphene oxide (GO) were fabricated by dissolving cellulose in a pre-cooled sodium hydroxide (NaOH)/urea solution and regenerated in sulphuric acid (H2SO4) coagulant. Initially, GO was found to disperse well in the cellulose solution due to intercalation with the cellulose; however, this cellulose-GO intercalation was disturbed during the regeneration process, causing agglomeration of GO in the RCF mixture. Agglomerated GO was confirmed at a higher GO content under a Dino-Lite microscope. The crystallinity index (CrI) and thermal properties of the RCFs increased with increasing GO loadings, up to 2 wt.%, and reduced thereafter. Cellulose-GO intercalation was observed at lower GO concentrations, which enhanced the crystallinity and thermal properties of the RCF-GO composite. It was shown that the GO exhibited antibacterial properties in the RCF-GO composite, with the highest bacterial inhibition against E. coli and S. aureus.

Surface modification for PDMS-based microfluidic devices.

This review focuses on advances reported from April 2009 to May 2011 in PDMS surface modifications for the application in microfluidic devices. PDMS surface modification techniques presented here include improved plasma and graft polymer coating, dynamic surfactant treatment, hydrosilylation-based surface modification and surface modification with nanomaterials such as carbon nanotubes and metal nanoparticles. Recent efforts to generate topographical and chemical patterns on PDMS are also discussed. The described surface modifications not only increase PDMS wettability, inhibit or reduce non-specific adsorption of hydrophobic species onto the surfaces in the act, but also result in the display of desired functional groups useful for molecular separations, biomolecular detection via immunoassays, cell culture and emulsion formation.

High-performance capillary electrophoretic separation of double-stranded oligonucleotides using a poly-(ethylpyrrolidine methacrylate-co-methylmethacrylate)-coated capillary.

Here we describe a capillary electrophoretic method for the separation of double-stranded oligonucleotides (ds-ODNs) ranging from 16-20 bp with 2 bp resolution using a low concentration of poly(ethylpyrrolidine methacrylate-co-methyl methacrylate) (PEPyM-co-PMMA) copolymer physically adsorbed to a capillary surface. Contrary to traditional DNA separations, we show that the ds-ODN with the highest molecular size eluted first and propose that this phenomena is due to a screening effect by the PEPyM-co-PMMA coating on the smaller ds-ODNs negative charge during elution. Key to the performance of this separation was a sample preparation time of less than 1 h and analysis time of 40 min. Repeatability of intraday migration time for the mixtures was typically < 1% relative standard deviation (n = 3). In addition, we demonstrate that the coating has an acceptable capillary lifetime of over 70 injections.