On-site Synthesis Research Articles

Tumor specific in situ synthesis of therapeutic agent for precision cancer therapy

BackgroundTraditional chemotherapeutic agents suffer from a lack of selectivity, poor targeting ability, and drug resistance. Developing tumor-specific therapies is crucial for precisely eliminating tumors while circumventing toxicity to normal tissues. Disulfiram (DSF), an FDA-approved drug for treating alcohol dependence, exhibits antitumor effect by forming complexes with copper ions (Cu(DDC)2). Here, we developed a Cu-doped polydopamine-based nanosystem (DSF@CuPDA-PEGM) to achieve in situ generation of toxic Cu(DDC)2.ResultsIn cancer cells with elevated H2O2 contents, CuPDA responsively degrades to release Cu ions and DSF, allowing on-site synthesis of Cu(DDC)2 with potent antitumor activity. DSF@CuPDA-PEGM exhibits excellent therapeutic efficacy against both drug-sensitive and drug-resistant cancer cells while minimizing toxicity to noncancerous cells. Moreover, DSF@CuPDA-PEGM promotes the immune response by inducing cancer cell immunogenic death, thereby augmenting anti-PD-1-based immune checkpoint blockade therapy.ConclusionA tumor-specifically degradable Cu-doped polydopamine-based nanosystem is developed to achieve in situ synthesis of antitumor compounds, providing a promising approach to precisely eliminate tumors and heighten chemo-immunotherapy.Graphical

Enhanced antibacterial activity of multi-walled carbon nanotubes loaded with hot-melt extruded Mulberry leaf silver nanoparticles

We suggest an antibacterial nanoplatform composed of multi-walled carbon nanotubes (MWCNTs) coated with mesoporous silica decorated with silver nanoparticles (AgNPs) via an N-[3-(trimethoxysilyl)propyl]ethylenediamine (TSD)-mediated method. In this system, the external mesoporous silica shell enhances the dispersion of MWCNTs to increase their contact area with bacterial cell walls. Meanwhile, the multiple mesoporous voids in the silica layer serve as a microreactor for on-site synthesis of small-sized and uniformly distributed AgNPs, thereby enhancing the antibacterial activity. Mulberry leaf (ML) extract was used to reduce Ag ions. In this process, hot melt extrusion (HME) was applied to increase the active ingredients of the ML extract. To prevent loss of ML components due to the heat and pressure applied during HME treatment, several excipients, including whey protein isolate, were mixed, and added. Compared with ML AgNPs and mesoporous silica-coated MWCNTs decorated with AgNPs (MWCNT-Ag), the MWCNT-Ag@ML (MWCNT-ML) exhibited stronger antibacterial against Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and methicillin-resistant S. aureus (MRSA). Therefore, this nano platform is a potential antibacterial agent and therapeutic agent for drug-resistant infections.

Next generation dry strength additives: Leveraging on-site synthesis to develop high performance glyoxalated polyacrylamides

Although glyoxalated polyacrylamides (gPAMs) have been described since the 1950s, the freedom to design new materials based on this chemistry has been limited by practical concerns; namely, a balance between solution concentration and material characteristics must be met to make the economics of gPAM strength additives work for the paper industry. For traditional “delivered” gPAMs, only a very narrow range of polyacrylamide molecular weights and compositions could be considered for glyoxalation. However, the development and successful implementation of automated reactor equipment that allows for the synthesis of gPAMs from glyoxal and polyacrylamide copolymers at the mill, known as “on-site” glyoxalation, obviates the shipping and stability concerns that have traditionally held back gPAM development. As such, on-site generators represent a platform that enables the glyoxalation of materials that would otherwise not have been suitable for use in a traditionally delivered gPAM product. These on-site generators therefore open new avenues for polymer design to allow for the creation of the next generation of strength additives. By leveraging the synthetic freedom of the on-site generators, a suite of high performance gPAMs has been designed, yielding materials that provide both exceptional strength and drainage performance in poor quality furnishes.

A Novel Process for the On-Site Preparation and Application of Polyferric Chloride (PFC) for Surface Water Treatment

This is the first study to describe a novel, patented process for the on-site synthesis and subsequent direct utilisation of Polyferric Chloride (PFC) at low Fe concentration dosing, which aims to facilitate the potential replacement of Polyaluminium Chloride (PAC) during surface water treatment (e.g., from reservoirs) for drinking water production. For this purpose, the PFC was synthesised and subsequently used as a coagulant in simulated surface water samples under different synthesis and coagulation/flocculation conditions, namely for different pre-hydrolysed Fe concentrations, pre-hydrolysis pH, coagulation pH, and flocculation times. The effectiveness of PFC was examined mainly in terms of total organic carbon (TOC) removal and the residual Fe concentration. The obtained results showed that the pre-hydrolysed Fe concentration at 0.5 ± 0.25%, pre-hydrolysis at pH 2.5 ± 0.25, coagulation at pH 5.5–7.0 and a flocculation time of 5 min could result in the highest TOC removal (i.e., residual values < 0.60 mg/L) and the lowest residual Fe concentration (<5 μg Fe/L), which is acceptable for a water quality assessment. These values are also substantially lower when compared to the respective TOC and residual metal concentrations using PAC (usually, the relevant obtained values are around TOC > 1 mg/L and Al > 50 μg/L).

Abstract 6202: 6-Phosphofructo-2-kinase in regulating DNA damage and repair in EGFR-driven lung cancer

Abstract Activating mutations of epidermal growth factor receptor (mutEGFR) are established drivers of lung tumor development, aggressiveness, and resistance to targeted therapies. While targeting mutEGFR with small-molecule tyrosine kinase inhibitors (EGFR-TKIs) dramatically improved progression-free survival in patients with non-small cell lung cancers (NSCLCs), intrinsic or acquired resistance to TKI therapy limits the duration of an effective response to TKIs. Alteration in DNA damage response (DDR) plays a vital role in genomic stability and cancer progression and helps cells to escape apoptosis, limiting the efficacy of targeted therapies. Interestingly, lung cancer cells exposed to EGFR-TKIs demonstrate attenuated glycolytic flux - one of the major providers of precursors for de novo nucleotide synthesis. A key regulator of glycolysis is the enzyme 6-phosphofructo-2-kinase (PFKFB3). PFKFB3 synthesizes fructose 2,6-bisphosphate (F2,6BP), which is a potent allosteric activator of the glycolytic rate-limiting enzyme, 6-phosphofructo-1-kinase (PFK1). Accordingly, F2,6BP controls flux throughout the entire glycolytic pathway. In preliminary studies, we provide evidence that PFKFB3 plays multiple roles in regulating the efficacy of DDR in lung cancer cells exposed to EGFR-TKIs. We show that PFKFB3 is required for EGFR-driven glucose metabolism, which provides a list of intermediates utilized in nucleotide synthesis. PFKFB3 regulates the expression of ribonucleotide reductase small subunit M2 (RRM2) of the ribonucleotide reductase (RNR), which is important for de novo onsite synthesis of deoxynucleotide triphosphate (dNTP) during DNA replication and repair. Moreover, a PFKFB3 inhibitor, PFK-158, increases oxidative stress in mutEGFR lung cancer cell lines and, as a result, promotes DNA damage. Importantly, we found that PFKFB3 plays an important role in the chromatin binding of scaffold proteins regulating the assembly of DNA repair complex. Our studies suggest that PFKFB3 plays an important role in DDR and provide a clear rationale to propose the use of PFKFB3 inhibitors in combination with EGFR inhibitors to increase the efficiency and/or overcome resistance to EGFR-TKIs. Citation Format: Nadiia Lypova, Lilibeth Lanceta, Susan Dougherty, Jason Chesney, Yoannis Imbert-Fernandez. 6-Phosphofructo-2-kinase in regulating DNA damage and repair in EGFR-driven lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6202.

SERS determination of sodium saccharin content on the tipping paper of cigarettes using AgNP substrates prepared with a USB-power supply device.

A novel magnetic agitating heater powered by a USB port has been developed and applied to synthesize silver colloid substrate for surface-enhanced Raman scattering (SERS) detection of sodium saccharin content on the tipping paper of cigarettes. The heater device allows the convenient synthesis of the Ag colloid, and the reaction can be completed on-site in 15 min under mild conditions. The on-site synthesis of SERS substrate effectively avoided the need for storage and concerns regarding the poor stability and short lifespan of colloid substrates. The results demonstrated that the substrate obtained with the device could achieve SERS detection of Rhodamine 6G (R6G) at as low as 10-9 mol L-1 while maintaining a stable intensity with a relative standard deviation (RSD) of 5.52% (n = 5). Using the prepared substrate at the optimal conditions, the limit of detection of sodium saccharin (SS) was 1 mg L-1. By introducing an internal standard KSCN, a linear relationship was observed between the relative intensity at 708 cm-1 and the concentration of the SS in a range of 20-100 mg L-1 (R2 = 0.98). With the developed method, SS was directly extracted from the cigarette paper by immersing it in water, and the extracted solution was subsequently detected. The quantitative spike-recoveries were in the range of 95.5-116.7%, with RSD between 2.3-12.6%. The whole detection procedure including the on-site substrate preparation, took only about 30 min. This work opens new avenues for colloidal synthesis, and the detection method of SS on the cigarette paper also holds great promise in food safety applications.

On-Site Synthesis and Characterizations of Atomically-Thin Nickel Tellurides with Versatile Stoichiometric Phases through Self-Intercalation.

Self-intercalation of native metal atoms in two-dimensional (2D) transition metal dichalcogenides has received rapidly increasing interest, due to the generation of intriguing structures and exotic physical properties, however, only reported in limited materials systems. An emerging type-II Dirac semimetal, NiTe2, has inspired great attention at the 2D thickness region, but has been rarely achieved so far. Herein, we report the direct synthesis of mono- to few-layer Ni-tellurides including 1T-NiTe2 and Ni-rich stoichiometric phases on graphene/SiC(0001) substrates under ultra-high-vacuum conditions. Differing from previous chemical vapor deposition growth accompanied with transmission electron microscopy imaging, this work combines precisely tailored synthesis with on-site atomic-scale scanning tunneling microscopy observations, offering us visual information about the phase modulations of Ni-tellurides from 1T-phase NiTe2 to self-intercalated Ni3Te4 and Ni5Te6. The synthesis of Ni self-intercalated NixTey compounds is explained to be mediated by the high metal chemical potential under Ni-rich conditions, according to density functional theory calculations. More intriguingly, the emergence of superconductivity in bilayer NiTe2 intercalated with 50% Ni is also predicted, arising from the enhanced electron-phonon coupling strength after the self-intercalation. This work provides insight into the direct synthesis and stoichiometric phase modulation of 2D layered materials, enriching the family of self-intercalated materials and propelling their property explorations.

Understanding the selectivity trend of water and sulfate (SO42−) oxidation on metal oxides: On-site synthesis of persulfate, H2O2 for wastewater treatment

Herein, H2O2 production activity via two-electron water oxidation reaction is identified on metal oxides (WO3, TiO2, and BiVO4) in acidic SO42− involving electrolyte, in which persulfate (PS) acts as an intermediate. A deep understanding of mechanism of PS production reaction on metal oxides has been realized based on rigorous crystal plane analysis of WO3 and TiO2 by combining density functional theory calculations and zeta potential measurements. It is found that the binding-energy of adsorbed hydroxyl groups on dominant exposed crystal plane of WO3 is lower than that of TiO2, leading to more SO42− adsorption sites at the surface of WO3. Consequently, the formation of PS on WO3 surface is more favorable. Meanwhile, both SO4−· and OH· were detected in the electrolyte by electron paramagnetic resonance measurement, which play a crucial role in organic pollutants wastewater treatment, with a degradation efficiency of 96% in 90 min for a typical pollutant norfloxacin. This work can provide a novel insight for understanding the selectivity of water and SO42− species oxidation on metal oxides, and open a new way for both on-site H2O2 production and advanced oxidation processes application.

Complex supramolecular tessellations with on-surface self-synthesized C60 tiles through van der Waals interaction.

Supramolecular tessellation with self-synthesized (C60)7 tiles is achieved based on a cooperative interaction between co-adsorbed C60 and octanethiol (OT) molecules. Tile synthesis and tiling take place simultaneously on a gold substrate leading to a two-dimensional lattice of (C60)7 tiles with OT as the binder molecule filling the gaps between the tiles. This supramolecular tessellation is featured with simultaneous on-site synthesis of tiles and self-organized tiling. In the absence of specific functional groups, the key to ordered tiling for the C60/OT system is the collective van der Waals (vdW) interaction among a large number of molecules. This bicomponent system herein offers a way for the artificial synthesis of 2D complex vdW supramolecular tessellations.

A modern baseline for the paired isotopic analysis of skin and bone in terrestrial mammals.

We present the isotopic discrimination between paired skin and bone collagen from animals of known life history, providing a modern baseline for the interpretation of archaeological isotopic data. At present, the interpretation of inter-tissue variation (Δ(skin–bone)) in mummified remains is based on comparisons with other archaeological material, which have attributed divergence to their contrasting turnover rates, with rapidly remodelling skin collagen incorporating alterations in environmental, cultural and physiological conditions in the months prior to death. While plausible, the lack of baseline data from individuals with known life histories has hindered evaluation of the explanations presented. Our analysis of a range of animals raised under a variety of management practices showed a population-wide trend for skin collagen to be depleted in 13C by –0.7‰ and enriched in 15N by +1.0‰ relative to bone collagen, even in stillborn animals. These results are intriguing and difficult to explain using current knowledge; however, on the basis of the findings reported here, we caution any results which interpret simply on differing turnover rates. We hypothesize that there may be a consistent difference in the routing of dietary protein and lipids between skin and bone, with potentially on-site synthesis of non-essential amino acids using carbon and nitrogen that have been sourced via different biochemical pathways.

Transforming C60 Molecules into Polyhedral Carbon Micro-Nano Shells for Electrochemically Producing H2O2 in Neutral Electrolytes.

The electrochemical production of hydrogen peroxide (H2O2) via the two-electron oxygen reduction reaction (ORR) can realize the customer-oriented onsite synthesis of H2O2 in a green and sustainable method. The ongoing challenge that needs to be solved is the fabrication of robust electrocatalysts of excellent performance. In this work, C60 was selected as a precursor due to its uniform structure and abundant pentagon rings. Thanks to the strong interaction between C60 and thiophene, after heteromolecule assembly in the liquid reaction and subsequent reconstruction of the carbon topological structure in solid calcination, C60 was successfully transformed into polyhedral carbon micro-nano shells (PCMNS) with an effective pore structure for the first time, which exhibited excellent capacity for production of H2O2 via two-electron ORR, especially in neutral media. In addition to the high onset potential (0.49 V vs reversible hydrogen electrode (RHE)) and low Tafel slope (72 mV dec-1), its selectivity reached >90% within the potential range of 0.30-0.45 V and maintained >80% after constant potential electrolysis for 10 h. The yield rate of H2O2 was 1102.5 mmol gcat-1 h-1, determined by an H-type electrolytic cell, which was one of the highest values of metal-free carbon-based ORR electrocatalysts ever reported. Such excellent two-electron ORR performance of PCMNS was attributed to its abundant accessible active sites and hierarchical pore structures.

Decoding of Oxygen Network Distortion in a Layered High-Rate Anode by In Situ Investigation of a Single Microelectrode.

Sluggish conversion reactions severely impair the rate capability for lithium storage, which is the main disadvantage of the conversion-type anode materials. Here, the microplatform based on a single microelectrode is designed and utilized for the fundamental understanding of the conversion reaction. The kinetic-favorable layered structure of the anode material is on-site synthesized in the microplatform. The in situ characterization reveals that introducing an oxygen network distortion in the layered oxide anode effectively circumvents the severe passivation of the electrode material by lithium oxide, thus leading to highly reversible conversion reactions. As a result, the high-rate capability of the conversion-type anode materials is realized. The on-site synthesis strategy is further applied in the large-scale synthesis of nanomaterials for lithium-ion batteries. As such, oxide nanorods with the layered structure are synthesized by a facile chemical strategy, showing high rate performance (574 mAh g-1 at 10 A g-1). This work unveils the beneficial effect of oxygen network distortion in the layered anode for conversion reactions over cycling, thus providing an alternative strategy to enhance the rate capability of conversion-type anodes for lithium storage.

Dynamic Genome Editing Using In Vivo Synthesized Donor ssDNA in Escherichia coli.

As a key element of genome editing, donor DNA introduces the desired exogenous sequence while working with other crucial machinery such as CRISPR-Cas or recombinases. However, current methods for the delivery of donor DNA into cells are both inefficient and complicated. Here, we developed a new methodology that utilizes rolling circle replication and Cas9 mediated (RC-Cas-mediated) in vivo single strand DNA (ssDNA) synthesis. A single-gene rolling circle DNA replication system from Gram-negative bacteria was engineered to produce circular ssDNA from a Gram-positive parent plasmid at a designed sequence in Escherichia coli. Furthermore, it was demonstrated that the desired linear ssDNA fragment could be cut out using CRISPR-associated protein 9 (CRISPR-Cas9) nuclease and combined with lambda Red recombinase as donor for precise genome engineering. Various donor ssDNA fragments from hundreds to thousands of nucleotides in length were synthesized in E. coli cells, allowing successive genome editing in growing cells. We hope that this RC-Cas-mediated in vivo ssDNA on-site synthesis system will be widely adopted as a useful new tool for dynamic genome editing.

Self-organized Ruthenium-Barium Core-Shell Nanoparticles on a Mesoporous Calcium Amide Matrix for Efficient Low-Temperature Ammonia Synthesis.

A low-temperature ammonia synthesis process is required for on-site synthesis. Barium-doped calcium amide (Ba-Ca(NH2 )2 ) enhances the efficacy of ammonia synthesis mediated by Ru and Co by 2 orders of magnitude more than that of a conventional Ru catalyst at temperatures below 300 °C. Furthermore, the presented catalysts are superior to the wüstite-based Fe catalyst, which is known as a highly active industrial catalyst at low temperatures and pressures. Nanosized Ru-Ba core-shell structures are self-organized on the Ba-Ca(NH2 )2 support during H2 pretreatment, and the support material is simultaneously converted into a mesoporous structure with a high surface area (>100 m2 g-1 ). These self-organized nanostructures account for the high catalytic performance in low-temperature ammonia synthesis.

Synthesis-free PET imaging of brown adipose tissue and TSPO via combination of disulfiram and 64CuCl2

PET imaging is a widely applicable but a very expensive technology. On-site synthesis is one important contributor to the high cost. In this report, we demonstrated the feasibility of a synthesis-free method for PET imaging of brown adipose tissue (BAT) and translocator protein 18 kDa (TSPO) via a combination of disulfiram, an FDA approved drug for alcoholism, and 64CuCl2 (termed 64Cu-Dis). In this method, a step-wise injection protocol of 64CuCl2 and disulfiram was used to accomplish the purpose of synthesis-free. Specifically, disulfiram, an inactive 64Cu ligand, was first injected to allow it to metabolize into diethyldithiocarbamate (DDC), a strong 64Cu ligand, which can chelate 64CuCl2 from the following injection to form the actual PET tracer in situ. Our blocking studies, western blot, and tissue histological imaging suggested that the observed BAT contrast was due to 64Cu-Dis binding to TSPO, which was further confirmed as a specific biomarker for BAT imaging using [18F]-F-DPA, a TSPO-specific PET tracer. Our studies, for the first time, demonstrated that TSPO could serve as a potential imaging biomarker for BAT. We believe that our strategy could be extended to other targets while significantly reducing the cost of PET imaging.

Microfluidics based Handheld Nanoparticle Synthesizer

Current study relates to the development of an electrical power-free, handheld microfluidic nanoparticle synthesizer for synthesis of uniform sized silver nanoparticles at room temperature. The synthesizer module consists of a custom designed microreactor and employs negative pressure based pumping mechanism for the electrical power free synthesis of metal nanoparticles. In order to realize a microreactor capable of on-site synthesis of monodisperse nanoparticles, optimization studies by bulk biosynthesis at varying ratios of the precursor and the reducing agent followed by UV-VIS absorption studies were performed to determine the appropriate mixing ratio. Later, a custom designed microfluidic micromixer was used to perform volumetric flow rate optimizations at the desired ratio using syringe pumps. From the knowledge of the precursor and reducing agent ratio and the flow rates, we modified the hydraulic resistance of micro-mixer inlets by varying the channel geometry to meet the optimized specifications leading to effective synthesis. The synthesized nanoparticles were characterized by UV-VIS spectroscopy, XPS, FTIR, EDS, HRTEM and SAED. The crystal lattice planes of 111] and 220] from the SAED pattern confirms the presence of silver nanoparticles. HRTEM study elucidates that the size of the synthesized nanoparticles is between 2 and 10 nm.

Safe, Selective, and High-Yielding Synthesis of Acryloyl Chloride in a Continuous-Flow System.

Acid chlorides are an important class of compounds and their high reactivity and instability has prompted us to develop a straightforward procedure for their synthesis with on-demand and on-site synthesis possibilities. The focus of this report is acryloyl chloride, mainly important for the acrylate and polymer industry. A continuous-flow methodology was developed for the fast and selective synthesis of the otherwise highly unstable acryloyl chloride. Three routes were investigated in a microreactor setup and all three can potentially be used for its production. The methodology was further expanded to the synthesis of other unstable acid chlorides by both the thionyl chloride and the oxalyl chloride mediated processes. The most sustainable method was the oxalyl chloride mediated procedure under solvent-free conditions, in which near-equimolar amounts of carboxylic acid and oxalyl chloride were used in the presence of catalytic amounts of DMF at room temperature. Within 1 to 3 min, nearly full conversions into the acid chlorides were achieved.

Onsite synthesis of thermally percolated nanocomposite for thermal interface material

To solve the problem of lack of thermal percolation in thermal interface materials (TIM), we propose a two-step synthesis method to realize thermally percolated nanofiber network in polymer matrix. First, by packing vapor grown carbon fibers (VGCFs) on top of aluminum heat sink and integrally sintering the whole material, the aluminum partially melts and connects the VGCF network, forming a continuous thermal path, i.e., realizing thermal percolation. Second, the pores in the hybrid network are filled by Silicone oil to obtain a polymer nanocomposite. The direct synthesis of VGCF-aluminum network on the heat sink (onsite synthesis) omits pasting process of the TIM, and thus, removes the restriction on the network morphology. By this onsite synthesis method, we reinforce thermal contact not only between the nanofibers but also between nanofibers and the heat sink. By testing the developed TIM for thermal contact to silicon surface, we demonstrate the potential to significantly reduce thermal contact resistance from what can be achieved by a conventional TIM.

Gold-nanoparticle, functionalized-porous-polymer monolith enclosed in capillary for on-column SERS detection

Gold nanoparticles functionalized porous polymer monoliths were developed via on-site synthesis method and enclosed in silica capillary as sensitive, uniform, and stable surface-enhanced Raman scattering (SERS) substrates.

Biomass-directed synthesis of 20 g high-quality boron nitride nanosheets for thermoconductive polymeric composites.

Electrically insulating boron nitride (BN) nanosheets possess thermal conductivity similar to and thermal and chemical stabilities superior to those of electrically conductive graphenes. Currently the production and application of BN nanosheets are rather limited due to the complexity of the BN binary compound growth, as opposed to massive graphene production. Here we have developed the original strategy "biomass-directed on-site synthesis" toward mass production of high-crystal-quality BN nanosheets. The strikingly effective, reliable, and high-throughput (dozens of grams) synthesis is directed by diverse biomass sources through the carbothermal reduction of gaseous boron oxide species. The produced BN nanosheets are single crystalline, laterally large, and atomically thin. Additionally, they assemble themselves into the same macroscopic shapes peculiar to original biomasses. The nanosheets are further utilized for making thermoconductive and electrically insulating epoxy/BN composites with a 14-fold increase in thermal conductivity, which are envisaged to be particularly valuable for future high-performance electronic packaging materials.