Multiple Stabilization Effects of Benzylhydrazine on Scalable Perovskite Precursor Inks for Improved Perovskite Solar Cell Production.

Perovskite precursor inks suffer various forms of degradation, such as iodide anion oxidation and organic cation breakdown, hindering reliable perovskite solar cell manufacturing. Here we report that benzylhydrazine hydrochloride (BHC) not only retards the buildup of iodine as previously reported but also prevents the breakdown of organic cations. Through investigating BHC and iodine chemical reactions, we elucidate protonation and dehydration mechanisms, converting BHC to harmless volatile compounds, thus preserving perovskite film crystallization and solar cell performance. This inhibition effect lasts nearly a month with minimal BHC, contrasting control inks without BHC where organic cations fully react in less than a week. This enhanced understanding, from additive stabilization to end products, promises improved perovskite solar cell production reliability.

Crosslinking Vinyl-Addition Polynorbornenes via Difunctional Diazirines to Generate Low Dielectric-Constant and Low Dielectric-Loss Thermosets.

Thermosets having low dielectric constant (Dk < 3) and low dielectric dissipation factor (Df < 0.003), high glass transition temperature (Tg > 150 °C), and good adhesion to copper are desirable for the low loss layers of the copper clad laminates (CCL) in next generation printed circuit boards. Three different difunctional diazirines are evaluated for both thermal and photochemical crosslinking of a high Tg vinyl-addition polynorbornene resin: poly(5-hexyl-1-norbornene) (poly(HNB)). The substrate polymer, crosslinked by the carbenes generated from the activated diazirines, forms thermosets with Dk < 2.3 and Df < 0.001 at 10 GHz depending on the identity of the diazirine and the loading. The Dk and Df values for one composition are stable for 1600 h at 125 °C in air and for 1400 h at 85 °C and 85% relative humidity, suggesting good long-term reliability of this thermoset. Adhesion of poly(HNB) to copper can be enhanced by priming the copper surface with a diazirine prior to high temperature lamination; peel strength values of greater than 7.5 N cm-1 are achieved. Negative-tone photopatterning of poly(HNB) with diazirines upon exposure to 365 nm light is demonstrated.

Diazirine-Functionalized Polyurethane Crosslinkers for Isocyanate-Free Curing of Polyol-Based Coatings.

Polyurethane coatings have strong material properties due to the hydrogen bonding inherent to the urethane groups. However, installing this urethane moiety usually requires curing through difficult-to-handle isocyanates. In this work, we show the development of a polyurethane-based crosslinker that can be used to formulate a one-component polyurethane coating with material properties similar to those of isocyanate-based polyurethane coatings. To achieve this, we used diazirine functionalities that generate carbenes upon heating, which react with alcohol functionalities in a polyol to generate a crosslinked network with a high storage modulus.

Safety Evaluation of a Prototypical Diazirine-Based Covalent Crosslinker and Molecular Adhesive.

bis-Diazirine reagents are increasingly being used as polymer crosslinkers, adhesives, and photopatterning agents in the materials sciences literature, but little effort has been made thus far to document their chemical safety profile. Here, we describe the results of a detailed toxicity assessment of a representative bis-diazirine. Safety was evaluated by a series of in vitro assays, which found the product to be non-mutagenic in bacterial tester strains TA98 and TA100, non-corrosive and non-irritating to skin, and requiring no classification for eye irritation or serious damage. While in vitro tests do not capture the integrated whole animal system, and thus cannot completely rule out the possibility of adverse responses, the results of this study suggest a desirable safety profile for bis-diazirine reagents and provide a solid foundation upon which to add in vivo assessment of safety risk and dose-response studies.

Functionalization of Polydimethylsiloxane with Diazirine-Based Linkers for Covalent Protein Immobilization.

Biomolecule attachment to solid supports is critical for biomedical devices, such as biosensors and implants. Polydimethylsiloxane (PDMS) is commonly used for these applications due to its advantageous properties. To enhance the biomolecule immobilization on PDMS, a novel technique is demonstrated using newly synthesized diazirine molecules for the surface modification of PDMS. This nondestructive process involves a reaction between diazirine molecules and PDMS through C-H insertion with thermal or ultraviolet activation. The success of the PDMS modification is confirmed by various surface characterization techniques. Bovine serum albumin (BSA) and immunoglobulin G (IgG) are strongly attached to the modified PDMS surfaces, and the amount of protein is quantified using iodine-125 radiolabeling. The results demonstrate that PDMS is rapidly functionalized, and the stability of the immobilized proteins is significantly improved with multiple types of diazirine molecules and activation methods. Confocal microscopy provides three-dimensional images of the distribution of immobilized IgG on the surfaces and the penetration of diazirine-based linkers through the PDMS substrate during the coating process. Overall, this study presents a promising new approach for functionalizing PDMS surfaces to enhance biomolecule immobilization, and its potential applications can extend to multimaterial modifications for various diagnostic and medical applications such as microfluidic devices and immunoassays with relevant bioactive proteins.

Synthesis of Graphene and Graphene Films with Minimal Structural Defects.

Graphene is a carbon material with extraordinary properties that has been drawing a significant amount of attention in the recent decade. High-quality graphene can be produced by different methods, such as epitaxial growth, chemical vapor deposition, and micromechanical exfoliation. The reduced graphene oxide route is, however, the only current approach that leads to the large-scale production of graphene materials at a reasonable cost. Unfortunately, graphene oxide reduction normally yields graphene materials with a high defect density. Here, we introduce a new route for the large-scale synthesis of graphene that minimizes the creation of structural defects. The method involves high-quality hydrogen functionalization of graphite followed by thermal dehydrogenation. We also demonstrated that the hydrogenated graphene synthesis route can be used for the preparation of high-quality graphene films on glass substrates. A reliable method for the preparation of these types of films is essential for the widespread implementation of graphene devices. The structural evolution from the hydrogenated form to graphene, as well as the quality of the materials and films, was carefully evaluated by Raman spectroscopy.

A Cleavable Crosslinking Strategy for Commodity Polymer Functionalization and Generation of Reprocessable Thermosets.

Covalently crosslinked polymeric materials, known as thermosets, possess enhanced mechanical strength and thermal stability relative to the corresponding uncrosslinked thermoplastics. However, the presence of covalent inter-chain crosslinks that makes thermosets so attractive is precisely what makes them so difficult to reprocess and recycle. Here, we demonstrate the introduction of chemically cleavable groups into a bis-diazirine crosslinker. Application of this cleavable crosslinker reagent to commercial low-functionality polyolefins (or to a small-molecule model) results in the rapid, efficient introduction of molecular crosslinks that can be uncoupled by specific chemical inputs. These proof-of-concept findings provide one potential strategy for circularization of the thermoplastic/thermoset plastics economy, and may allow crosslinked polyolefins to be manufactured, used, reprocessed, and re-used without losing value. As an added benefit, the method allows the ready introduction of functionality into non-functionalized commodity polymers.

An Economical and Scalable Method to Synthesize Graphitic-Like Films.

An economical and facile method to synthesize a precursor for carbon films and materials has been developed. This precursor can be easily coated onto substrates without binder reagents and then converted into a graphitic-like structure after mild thermal treatment. This approach potentially allows the coating of glass surfaces of different shapes and forms, such as the inside of a glass tube, for instance. The precursor consists of tetrahedral halocarbyne units which randomly combine through single electron transfer with organometallic compounds to create a poly(carbyne)-like polymeric material. Advanced characterization tools reveal that the synthesized product (poly(halocarbyne) or PXC, where X indicate the presence of halogens, is composed mostly of carbon, hydrogen, and a variable percentage of residual halocarbon groups. Therefore, it possesses good solubility in organic solvents and can be coated on any complex substrate. The coated PXC material produced here was annealed under mild conditions, leading to the production of a graphitic-like film on a glass substrate. The chemical homogeneity of the carbon material of the film was confirmed by Raman spectroscopy.

Light-Induced Anti-Bacterial Effect Against Staphylococcus aureus of Porphyrin Covalently Bonded to a Polyethylene Terephthalate Surface.

Antimicrobial photodynamic inactivation represents a promising and potentially greener alternative to conventional antimicrobials, and a solution for multidrug-resistant strains. The current study reports the development and characterization of tetra-substituted diazirine porphyrin covalently bonded to polyethylene terephthalate (PET) and its use as an antimicrobial surface. The diazirine moiety on the porphyrin was activated using a temperature of 120 °C, which initiated a C-H insertion mechanism that irreversibly functionalized the PET surface. Activation of the surface with white LED light in phosphate-buffered saline (PBS) led to singlet oxygen generation, which was detected via the degradation of 9,10-anthracenediylbis(methylene)dimalonic acid (ADMA) over time. The bactericidal effect of the 1O2-producing surface against Staphylococcus aureus was determined qualitatively and quantitatively. The growth of the pathogen beneath porphyrin-functionalized PET coupons was reduced; moreover, the PET coupons resulted in a 1.76-log reduction in cell counts after exposure to white LED light for 6 h. This is a promising material and platform for the development of safer antimicrobial surfaces, with applications in healthcare, food packaging, marine surfaces, and other surfaces in the environment.

Improvements in Drug-Delivery Properties by Co-Encapsulating Curcumin in SN-38-Loaded Anticancer Polymeric Nanoparticles.

SN-38 is an immensely potent anticancer agent although its use necessitates encapsulation to overcome issues of poor solubility and stability. Since SN-38 is a notoriously challenging drug to encapsulate, new avenues to increase encapsulation efficiency in polymer nanoparticles (PNPs) are needed. In this paper, we show that nanoprecipitation with curcumin (CUR) increases SN-38 encapsulation efficiencies in coloaded SN-38/CUR-PNPs based on poly(ε-caprolactone)-block-poly(ethylene glycol) (PCL-b-PEG) by up to a factor of 10. In addition, we find a dramatic decrease in PNP polydispersities, from 0.34 to 0.07, as the initial CUR-to-polymer ratio increases from 0 to 10, with only a modest increase in PNP size (from 40 to 55 nm). Compared to coloaded PNP formation using nanoprecipitation in the bulk or in a gas-liquid, a two-phase microfluidic reactor shows similar trends with respect to CUR content, although improvements in SN-38 encapsulation efficiencies both with and without CUR are found using the microfluidic method. Additional precipitation studies without copolymer suggest that CUR increases the dispersion of SN-38 in the solvent medium of micelle formation, which may contribute to the observed encapsulation enhancement. Cytotoxicity studies of unencapsulated SN-38/CUR mixtures show that addition of CUR does not significantly affect SN-38 potency against either U87 (glioblastoma) or A204 (rhabdomyosarcoma) cell lines. However, we find significant differences in the potencies of SN-38/CUR-PNP formulations depending on initial CUR amounts, with an optimized formulation showing subnanomolar cytotoxicity against A204 cells, significantly more potent than either free SN-38 or PNPs containing only SN-38.

A Chan-Evans-Lam approach to trisubstituted vinyl ethers.

Trisubstituted vinyl ethers were accessed via Chan-Evans-Lam coupling of vinyl trifluoroborates and primary aliphatic alcohols. This approach complements prior methods that required the use of neat liquid alcohol coupling partners. A palladium-catalyzed redox-relay Heck reaction was used to convert several vinyl ethers into aldehyde-functionalized 1,3-dihydroisobenzofurans.

Structure-function relationships in aryl diazirines reveal optimal design features to maximize C-H insertion.

Diazirine reagents allow for the ready generation of carbenes upon photochemical, thermal, or electrical stimulation. Because carbenes formed in this way can undergo rapid insertion into any nearby C-H, O-H or N-H bond, molecules that encode diazirine functions have emerged as privileged tools in applications ranging from biological target identification and proteomics through to polymer crosslinking and adhesion. Here we use a combination of experimental and computational methods to complete the first comprehensive survey of diazirine structure-function relationships, with a particular focus on thermal activation methods. We reveal a striking ability to vary the activation energy and activation temperature of aryl diazirines through the rational manipulation of electronic properties. Significantly, we show that electron-rich diazirines have greatly enhanced efficacy toward C-H insertion, under both thermal and photochemical activation conditions. We expect these results to lead to significant improvements in diazirine-based chemical probes and polymer crosslinkers.

Flexible polyfluorinated bis-diazirines as molecular adhesives.

Motivated by a desire to develop flexible covalent adhesives that afford some of the same malleability in the adhesive layer as traditional polymer-based adhesives, we designed and synthesized two flexible, highly fluorinated bis-diazirines. Both molecules are shown to function as effective crosslinkers for polymer materials, and to act as strong adhesives when painted between two polymer objects of low surface energy, prior to thermal activation. Data obtained from lap-shear experiments suggests that greater molecular flexibility is correlated with improved mechanical compliance in the adhesive layer.

Efficient purification of the diarylheptanoid oregonin from red alder (Alnus rubra) leaves and bark combining aqueous extraction, spray drying and flash-chromatography.

INTRODUCTION: The diarylheptanoid xyloside oregonin ((5S)-1,7-bis(3,4-dihydroxyphenyl)-5-(ß-d-xylopyranosyloxy)-heptan-3-one) has significant medicinal potential and is found at high concentration in leaves and bark of red alder (Alnus rubra). OBJECTIVES: To establish inexpensive and easily scaled methods for the extraction and purification of oregonin from timber by-products. METHODS: We developed a method combining aqueous extraction with spray drying of red alder extract into a powder, thus reducing the need for organic solvents used in traditional Soxhlet extraction or in solvent partitioning. Flash chromatography was utilised to purify oregonin from crude spray-dried alder extract. RESULTS: Crude spray-dried alder extract was comprised of an average of 9% of the diarylheptanoid compound oregonin. Less than 10% thermal degradation of oregonin was observed using extraction temperatures between 25°C and 50°C, followed by spray drying. The structure of purified oregonin was validated using high-performance liquid chromatography (HPLC), mass spectrometry (MS), ultraviolet spectroscopy (UV), and nuclear magnetic resonance (NMR). CONCLUSION: The developed method was robust, repeatable, and yielded purified oregonin of greater than > 95% purity (average of 95.8%). Our analysis represents the most complete NMR characterisation of oregonin reported to date.

Development and Field Validation of Lidocaine-Loaded Castration Bands for Bovine Pain Mitigation.

Castration is among the most common management procedures performed in the dairy and beef cattle industries and is mainly performed by surgery or elastic banding. Despite the various benefits of castration, all methods produce pain and distress. Castration by banding is simple, inexpensive, produces fewer complications, and can be performed in a high-throughput manner. Because lidocaine, a local anesthetic, can be delivered to trauma sites topically, we have formulated lidocaine-loaded castration bands (LLBs) to deliver local pain relief to calves during banded castration. The initial lidocaine content of three band types developed was between 80 and 200 mg per band. The transfer kinetics of lidocaine into tissue was determined in vitro, indicating a rapid release for the first 30 min, followed by a slow release lasting at least 48 h. Furthermore, the lidocaine delivery and pain mitigation effects of these LLBs were compared to standard lidocaine injections in vivo. Field studies indicated that LLBs performed at least as well as lidocaine injections for short-term lidocaine delivery into tissues and pain mitigation. Moreover, LLBs significantly outperformed lidocaine injections for long-term delivery and pain mitigation. The concentrations of lidocaine in the LLB-treated tissue samples were generally in the range of 0.5-3.5 mg of lidocaine per gram of tissue and were overall highest after 6 h. Lidocaine-loaded elastration bands deliver therapeutic quantities of lidocaine into scrotal tissues over a period of at least seven days in cattle. This approach would provide long-term pain mitigation to the animals and, by avoiding surgery or the administration of injections, would also decrease the time and handling costs for the producer.

Discovery of salicyl benzoate UDP-glycosyltransferase, a central enzyme in poplar salicinoid phenolic glycoside biosynthesis.

The salicinoids are anti-herbivore phenolic glycosides unique to the Salicaceae (Populus and Salix). They consist of a salicyl alcohol glucoside core, which is usually further acylated with benzoic, cinnamic or phenolic acids. While salicinoid structures are well known, their biosynthesis remains enigmatic. Recently, two enzymes from poplar, salicyl alcohol benzoyl transferase and benzyl alcohol benzoyl transferase, were shown to catalyze the production of salicyl benzoate, a predicted potential intermediate in salicinoid biosynthesis. Here, we used transcriptomics and co-expression analysis with these two genes to identify two UDP-glucose-dependent glycosyltransferases (UGT71L1 and UGT78M1) as candidate enzymes in this pathway. Both recombinant enzymes accepted only salicyl benzoate, salicylaldehyde and 2-hydroxycinnamic acid as glucose acceptors. Knocking out the UGT71L1 gene by CRISPR/Cas9 in poplar hairy root cultures led to the complete loss of salicortin, tremulacin and tremuloidin, and a partial reduction of salicin content. This demonstrated that UGT71L1 is required for synthesis of the major salicinoids, and suggested that an additional route can lead to salicin. CRISPR/Cas9 knockouts for UGT78M1 were not successful, and its in vivo role thus remains to be determined. Although it has a similar substrate preference and predicted structure as UGT71L1, it appears not to contribute to the synthesis of salicortin, tremulacin and tremuloidin, at least in roots. The demonstration of UGT71L1 as an enzyme of salicinoid biosynthesis will open up new avenues for the elucidation of this pathway.

A broadly applicable cross-linker for aliphatic polymers containing C-H bonds.

Addition of molecular cross-links to polymers increases mechanical strength and improves corrosion resistance. However, it remains challenging to install cross-links in low-functionality macromolecules in a well-controlled manner. Typically, high-energy processes are required to generate highly reactive radicals in situ, allowing only limited control over the degree and type of cross-link. We rationally designed a bis-diazirine molecule whose decomposition into carbenes under mild and controllable conditions enables the cross-linking of essentially any organic polymer through double C-H activation. The utility of this molecule as a cross-linker was demonstrated for several diverse polymer substrates (including polypropylene, a low-functionality polymer of long-standing challenge to the field) and in applications including adhesion of low-surface-energy materials and the strengthening of polyethylene fabric.

In Vitro Assessment of Putative PD-1/PD-L1 Inhibitors: Suggestions of an Alternative Mode of Action.

The programmed cell death protein 1 (PD-1) signaling axis is among the most important therapeutic targets in modern oncology. Aurigene Discovery Technologies Ltd. (Aurigene) has patented a series of peptidomimetic small molecules derived from the PD-1 protein sequence for use in targeting the interaction between PD-1 and its ligand, PD-L1. We evaluated three of Aurigene's most potent compounds in SPR binding assays. Our results showed that these compounds-each of which is known to be potently effective in a splenocyte recovery assay-do not directly inhibit the PD-1/PD-L1 interaction nor do they appear to bind to either of the constituent proteins, indicating that another mechanism is at play. As a result of these studies and upon consideration of structural features within the PD-1/PD-L1 complex, we hypothesize that the Aurigene molecules may interact with a currently unknown protein capable of regulating the PD-1 axis.