Purification Of Complex Research Articles

Development of Komagataella phaffii as a cell factory for efficient de novo production of β-caryophyllene.

β-Caryophyllene is a natural bicyclic sesquiterpene found in a large number of plants around the world. It has anti-inflammatory, anticancer and analgesic biological activities associated with its important medicinal value, and has also attracted attention in the field of bioenergy with high energy density. Due to the low amount of β-caryophyllene in plants and complex purification process, microbial biosynthesis is considered as a promising alternative for the industrial development of β-caryophyllene. Komagataella phaffii has a robust transcriptional regulatory system and has many advantages in protein expression, high-density culture, making it suitable for large-scale industrial production. However, there are no systematic studies on the efficient biosynthesis of β-caryophyllene in K. phaffii. In this study, firstly, farnesyl diphosphate synthase ERG20 and β-caryophyllene synthase AaCPS were fused and expressed with different linkers. Secondly, we enhanced the mevalonate pathway and inhibited the branch pathway. At last, the copy number of ERG20-(PA)5-AaCPS were adjusted for the biosynthesis of β-caryophyllene, a highly efficient β-caryophyllene production strain AaCPS16 was constructed. AaCPS16 could produce 136.4mg/L β-caryophyllene in shake flask level, which was 37 times higher than the initial strain AaCPS1. To the best of our knowledge, this is the first report of caryophyllene biosynthesis in Komagataella phaffii. This established a good foundation for the synthesis of sesquiterpenes in K. phaffii.

Production of a new variant of soluble trimer Env of HIV-1 CRF63_02A6 SOSIP.664

Introduction. Obtaining stabilized recombinant HIV-1 Env trimers that have a close to the native conformation is one of the directions in the field of development of vaccines against HIV-1. The aim of the study was to obtain and characterize the stabilized trimer Env of HIV-1 SOSIP.664 based on the circulating genetic variant of the recombinant form CRF63_02A6. Materials and methods. Bioinformatics resources were used to design the trimer Env gene based on the HIV-1 recombinant genetic variant CRF63_02A6. The designed gene was synthesized and cloned as part of an integration plasmid vector. A stable producer of trimer Env was obtained by transfection of the CHO-K1 cell line using the developed plasmid vector. Purification of the protein complex was performed using affinity chromatography and gel filtration. Antigenic properties of trimer Env were studied using immunochemical analysis using broadly neutralizing HIV-1 monoclonal antibodies (bnAbs). Results. A new variant of the stabilized trimer Env of the surface glycoprotein of the recombinant form CRF63_02A6 HIV-1 SOSIP.664 was designed, including additional stabilizing modifications. Based on the CHO-K1 cell line, a stable producer was obtained and a purification protocol for the designed trimer Env was developed. It was found that the trimer Env CRF63_02A6 SOSIP.664 is effectively recognized by bnAbs 2G12, VRC01 and PGT126. Conclusion. The obtained results indicate the prospects for further study of the structural features of the trimer Env CRF63_02A6 SOSIP.664, as well as its immunogenicity and the possibility of using it as a vaccine antigen.

Facile Synthesis of Fluorescent Carbon Quantum Dots with High Product Yield Using a Solid-Phase Strategy.

The liquid-phase method is the most commonly utilized strategy for synthesizing fluorescent carbon quantum dots (CQDs). However, the liquid-phase synthesis of CQDs faces challenges such as low yield, complex purification, and the use of toxic solvents, which limit large-scale production and practical applications. In this study, fluorescent CQDs with a high product yield of 78% were synthesized using glucose as a carbon source through a green and facile one-step solid-phase approach, without solvents or post-treatment. A systematic study of the structure and fluorescence properties of the synthesized CQDs was conducted using various characterization techniques. The results indicated that the mean size of obtained CQDs was 4.1 nm, and that their surface had abundant oxygen-containing functional groups, resulting in favorable water solubility. The synthesized CQDs exhibited excitation-dependent fluorescence, with optimal excitation and emission wavelengths at 358 and 455 nm, respectively. Additionally, the CQDs solution showed bright blue fluorescence under 365 nm UV light, with a quantum yield of 6.21% and a fluorescence lifetime of 3.02 ns. This study offers valuable insights into the green and efficient synthesis of fluorescent CQDs powder.

Urease-powered nanomotor probe for PCA3 ultrasensitive detection in human urine and controllable imaging in live cells

Prostate cancer (PCa) is the second-most common cancer in men. Recent researches indicate that prostate cancer antigen 3 (PCA3) is highly specific for the early screening of PCa. However, conventional techniques for PCA3 detection still remain significant challenges, including complex purification process, long detection time and poor sensitivity. Herein, we present urease-powered Janus nanomotors (JUNMs) to assist the nucleic-acid probe for sensitive urine analysis of prostate cancer (PCa)-related PCA3 and precise in situ imaging of PCa cells. The nanoprobes were synthesized by immobilizing urease and hairpin DNA asymmetrically on the Janus Fe3O4/Au nanoparticles (JUNMs-hDNA). JUNMs-hDNA nanomotors exhibited active motion in the presence of urea to efficiently recognize and capture the PCA3. As a result, the nanoprobes revealed a good linear relationship to PCA3 in the range of 1-100 pM. The total detection time was within 11minutes with a limit of detection (LOD) of 0.6 pM. Subsequently, JUNMs-hDNA nanoprobes have also been successfully applied in the detection of PCA3 in human urine samples. In addition, the JUNMs-hDNA nanoprobes also effectively captured PCa cells and enabled fluorescence imaging, which provided a highly sensitive sensing and imaging platform for PCA3 in body fluids and cellular level. Thus, this strategy offers a new perspective and approach for early, non-invasive and precise clinical diagnosis of PCa.

Preparation of magnesium potassium phosphate cement from magnesite tailings under lower calcination temperature

The identification of a more economical source of magnesium, coupled with reduced preparation costs, is essential for enhancing the competitiveness of magnesium potassium phosphate cement (MKPC) production. This study explores the utilization of magnesite tailings, waste materials produced during the mining and ore dressing process of magnesite, as raw materials for the preparation of low-cost dead-burnt magnesium oxide (MgO-T) at comparatively low calcination temperatures. The research demonstrated that impurities, specifically silicon and calcium, formed compounds with MgO during calcination, leading to the encapsulation of certain MgO particles. This encapsulation reduced both the specific surface area and hydration activity of the MgO. Despite the calcination temperature of 1100 °C being significantly lower than that of commercial dead-burnt MgO (MgO-C) at 1700 °C, MgO-T exhibited lower activity compared to MgO-C, which rendered it more suitable for the production of MKPC. Under optimal conditions, characterized by a molar ratio of magnesium to phosphorus (M/P) of 3, a borax retarder content of 15 %, and a water-cement ratio of 0.15, the MKPC prepared using MgO-T demonstrated an initial setting time exceeding 60 min and achieved a compressive strength of up to 79.6 MPa. XRD and microstructural analyses revealed that struvite-K was the primary strength phase; however, the porosity of the specimens had a more pronounced effect on strength. By producing lower activity MgO at reduced calcination temperatures, this method obviates the need for complex purification processes when utilizing magnesite tailings, thereby decreasing the costs associated with MKPC preparation and presenting promising applications for the future.

Label-Free Detection of Programmed Death-Ligand 1 for Prognosis Using a Truncated Aptamer and SYBR Green I.

The rapid and accurate detection of programmed death-ligand 1 (PD-L1) expression is of great value in the diagnosis and treatment of tumors. ELISA-based traditional method is the gold standard for protein detection, but there are still some shortcomings, especially the antigen-antibody dependence, greatly increased the detection time and cost. This work constructed a label-free fluorescent probe for rapid and sensitive detection of PD-L1 using a truncated aptamer as recognition molecules and double-stranded DNA specific dyes (SYBR Green I) as signal units. After a series of optimization conditions, this probe has good detection capability for PD-L1 in buffer solution with the detection limit as low as 0.68 ng/mL. Due to the specific recognition ability of aptamer and target, this method also has good selectivity for PD-L1 detection. The recovery of PD-L1 in human serum samples ranges from 86.20 to 96.36%. Compared with other methods, this strategy does not need to be marked, and does not need other complex design and purification process, but simple operation process and strong anti-interference ability. The whole detection process can be completed within 20min and has good application prospect. This work will provide reference for drug dosage and prognosis evaluation of specific tumor therapy.

From waste to an added-value commodity: challenges in valorization opportunity of tin-residual sand for silica-based industry

Abstract Extensive tin mining activities produce sand piles with high silica, leaving vast environmental and economic responsibilities. The circular economy implementation can address these problems by valorizing quartz sand as industrial sand. Therefore, the study assesses the quartz sand left over from tin mining and processing activities on Bangka Island to become a valuable product within the CPQvA Framework consisting of classification (C), potential utilization (P), quantity and viability (Qv), and application (A). The criteria comprise 12 weighted questions to define the criticality indices categorized as easy, moderate, and difficult. With complex purification technology, quartz sand can be used as silica sand for ceramics, foundry mould, refractory bricks, glass, and solar cells. The quantity is large and feasible for a factory scale with expensive investment and operational costs. The product performance shows good quality from a technical aspect. Due to impurities content and its complex purification technology, the valorization of quartz sand produces a difficult criticality index for the solar cells, a moderate criticality index for the glass and ceramics industry, and an easy criticality index for the glaze, refractory brick, and foundry mold industries. Therefore, proven and efficient purification technology is crucial for further research.

Advances in Inorganic Foam Materials Fabricated Via Blowing Strategy: A Comprehensive Review.

Two-dimensional (2D) materials with excellent properties and widespread applications have been explosively investigated. However, their conventional synthetic methods exhibit concerns of limited scalability, complex purification process, and incompetence of prohibiting their restacking. The blowing strategy, characterized by gas-template, low-cost, and high-efficiency, presents a valuable avenue for the synthesis of 2D-based foam materials and thereby addresses these constraints. Whereas, its comprehensive introduction has been rarely outlined so far. This review commences with a synopsis of the blowing strategy, elucidating its development history, the statics and kinetics of the blowing process, and the choice of precursor and foaming agents. Thereafter, we dwell at length on across-the-board foams enabled by the blowing route, like BxCyNz foams, carbon foams, and diverse composite foams consisting of carbon and metal compounds. Following that, a wide-ranging evaluation of the functionality of the foam products in fields such as energy storage, electrocatalysis, adsorption, etc. is discussed, revealing their distinctive strength originated from the foam structure. Finally, after concluding the current progress, we provide some personal discussions on the existing challenges and future research priorities in this rapidly developing method.

Advances in the synthesis of cytidine-5'-diphosphate choline

Cytidine-5'-diphosphate choline (CDP-choline) plays a crucial role in the formation of the phospholipid bilamolecular layer in cell membranes and the stabilization of the neurotransmitter system, acting as a precursor to phosphatidylcholine and acetylcholine. CDP-choline has been found effective in treating functional and consciousness disorders resulting from brain injury, Parkinson's disease, depression and glaucoma, and other conditions. As such, CDP-choline is widely utilized in clinical medicine and health care products. The conventional chemical synthesis process of CDP-choline is gradually being replaced by biosynthesis due to the expensive and toxic reagents involved, the production of various by-products, and the high cost of industrial production. Biosynthesis of CDP-choline offers two strategies: microbial fermentation and biocatalysis. Microbial fermentation utilizes inexpensive raw materials but results in a relatively low conversion rate and requires a complex separation and purification process. Biocatalysis, on the other hand, involves two stages: the growth of a living "catalyst" and the conversion of the substrate. Although the synthetic process in biocatalysis is more complex, it offers a higher conversion ratio, and the downstream processing technique for extraction is relatively less costly. Consequently, biocatalysis is currently the primary strategy for the industrial production of CDP-choline. This review aims to summarize the progress made in both chemical synthesis and biosynthesis of CDP-choline, with particular focus on the metabolic pathway and the synthetic processes involved in biocatalysis, in order to provide insights for the industrial production of CDP-choline.

Purification and characterization of recombinant human superoxide dismutase integrated with resilin-like polypeptide

Human superoxide dismutase (hSOD1) plays an important role in the aerobic metabolism and free radical eliminating process in the body. However, the production of existing SOD faces problems such as complex purification methods, high costs, and poor product stability. This experiment achieved low-cost, rapid, and simple purification of hSOD1 through ammonium sulfate precipitation method and heat resistance of recombinant protein. We constructed a recombinant protein hSOD1-LR containing a resilin-like polypeptide tag and expressed it. The interest protein was purified by ammonium sulfate precipitation method, and the results showed that the purification effect of 1.5 M (NH4)2SO4 was the best, with an enzyme activity recovery rate of 80 % after purification. Then, based on its thermal stability, further purification of the interest protein at 60 °C revealed a purification fold of up to 24 folds, and the purification effect was similar to that of hSOD1-6xHis purified by nickel column affinity chromatography. The stability of hSOD1-LR showed that the recombinant protein hSOD1-LR has better stability than hSOD-6xHis. hSOD1-LR can maintain 76.57 % activity even after 150 min of reaction at 70 °C. At same time, hSOD1-LR had activity close to 80 % at pH < 5, indicating good acid resistance. In addition, after 28 days of storage at 4 °C and 40 °C, hSOD1-LR retained 92 % and 87 % activity, respectively. Therefore, the method of purifying hSOD1-LR through salt precipitation may have positive implications for the study of SOD purification.

An immobilized antibody-based affinity grid strategy for on-grid purification of target proteins enables high-resolution cryo-EM

In cryo-electron microscopy (cryo-EM), sample preparation poses a critical bottleneck, particularly for rare or fragile macromolecular assemblies and those suffering from denaturation and particle orientation distribution issues related to air-water interface. In this study, we develop and characterize an immobilized antibody-based affinity grid (IAAG) strategy based on the high-affinity PA tag/NZ-1 antibody epitope tag system. We employ Pyr-NHS as a linker to immobilize NZ-1 Fab on the graphene oxide or carbon-covered grid surface. Our results demonstrate that the IAAG grid effectively enriches PA-tagged target proteins and overcomes preferred orientation issues. Furthermore, we demonstrate the utility of our IAAG strategy for on-grid purification of low-abundance target complexes from cell lysates, enabling atomic resolution cryo-EM. This approach greatly streamlines the purification process, reduces the need for large quantities of biological samples, and addresses common challenges encountered in cryo-EM sample preparation. Collectively, our IAAG strategy provides an efficient and robust means for combined sample purification and vitrification, feasible for high-resolution cryo-EM. This approach holds potential for broader applicability in both cryo-EM and cryo-electron tomography (cryo-ET).

Exploring the Interactions at the Interface: Tailoring Carbazole‐Based Self‐Assembled Molecules with Varying Functional Groups for Enhancing the Performance of Inverted Perovskite Solar Cells

Four different carbazole‐based self‐assembled molecules (SAMs) with different terminal groups have been designed and synthesized as hole‐selective contacts for inverted perovskite solar cells to investigate their interfacial interactions and, consequently, the performance of the devices. Using the carbazole core as a reference, the effect of the thiophen‐2‐yl phenyl, or the hydroxymethyl phenyl attached to the core through a phenyl moiety, with that of the thiophene directly linked to the carbazole is compared. These new SAMs have been successfully synthesized using cost‐effective starting materials and a straightforward synthetic method, eliminating the need for expensive and complex purification processes. Subsequently, they have been applied as efficient hole‐selective contact in inverted perovskite solar cells, leading to an outstanding power conversion efficiency of 19.67% in the case of SAM5, containing a carbazole‐core substituted with double 2‐phenylthiophene side arms as functional group. The detailed characterization of the interface and the charge kinetics has allowed to determine the effect of each substituent.

Sensitive Detection and Differentiation of Biologically Active Ricin and Abrin in Complex Matrices via Specific Neutralizing Antibody-Based Cytotoxicity Assay.

Ricin and abrin are highly potent plant-derived toxins, categorized as type II ribosome-inactivating proteins. High toxicity, accessibility, and the lack of effective countermeasures make them potential agents in bioterrorism and biowarfare, posing significant threats to public safety. Despite the existence of many effective analytical strategies for detecting these two lethal toxins, current methods are often hindered by limitations such as insufficient sensitivity, complex sample preparation, and most importantly, the inability to distinguish between biologically active and inactive toxin. In this study, a cytotoxicity assay was developed to detect active ricin and abrin based on their potent cell-killing capability. Among nine human cell lines derived from various organs, HeLa cells exhibited exceptional sensitivity, with limits of detection reaching 0.3 ng/mL and 0.03 ng/mL for ricin and abrin, respectively. Subsequently, toxin-specific neutralizing monoclonal antibodies MIL50 and 10D8 were used to facilitate the precise identification and differentiation of ricin and abrin. The method provides straightforward and sensitive detection in complex matrices including milk, plasma, coffee, orange juice, and tea via a simple serial-dilution procedure without any complex purification and enrichment steps. Furthermore, this assay was successfully applied in the unambiguous identification of active ricin and abrin in samples from OPCW biotoxin exercises.

Optimization of ultrasonic pretreatment and analysis of chlorogenic acid in potato leaves

Chlorogenic acid (CA) is an effective ingredient that can strengthen immunity during following the COVID-19 era. The current cost of CA is high owing to its complex purification process and low yield (approximately 2%). In this study, a one-step path orthogonal experiment was designed based on the results from Gauss calculation, which consisted of acidity, coordination, and hydrolysis in molecules. The optimized extraction conditions were 60 ℃, 60 min, 1:20 liquid ratio, and 40% ethanol in a nitrogen atmosphere controlled using a device of our own design, which led to CA yields of up to 6.35% from potato leaves. The purified CA was analyzed using high-performance liquid chromatography, thin-layer chromatography, ultraviolet–visible spectroscopy, and molecular fluorescence. This accurate and reproducible method can not only be used to obtain high yields of CA but can also be used for the quality control of active plant products and their isomers.

Multiple strategies to improve extracellular secretion and activity of feruloyl esterase

Feruloyl esterase has a wide range of applications, but there are still problems with low enzyme yield and activity, and complex purification steps. Our previous research found Lactobacillus amylovorus feruloyl esterase could be secreted extracellular in Escherichia coli. In this study, multiple strategies were implemented to maximize the extracellular production of feruloyl esterase with improved activity in E. coli. Firstly, codon-optimized feruloyl esterase was obtained based on the preference of E. coli, resulting in 41.97 % increase in extracellular secretion. Furthermore, by cascading T7 promoters, replacing the 5′ UTR, randomly mutating the N-terminal sequence, and co-expressing secretory cofactors, the extracellular secretion was increased by 36.46 %, 31.25 %, 20.66 % and 25.75 %, respectively. Moreover, the feruloyl esterase were mutated to improve the substrate affinity and activity. The catalytic efficiency of Fae-Q134T and Fae-Q198A increased by 4.62-fold and 5.42-fold. Combining above strategies, extracellular feruloyl esterase activity was increased from 2013.70 U/L to 10,349.04 U/L. These results indicated that the activity and yield of feruloyl esterase secreted by E. coli were significantly increased, which laid a foundation for its industrial application.

WASTEWATER TREATMENT OF COAL TAR PROCESSING PROCESS FROM PHENOL

The coal tar processing process provides the chemical industry with valuable products: naphthalene, cresol, xylenols, phenol, technical oils and lubricants. The most significant product of coal tar processing is anthracene, which is a raw material for the production of carbon black. The prospects for the implementation of the coal tar processing process entail a danger to the environment and public health.Wastewater formed during the processing process has a complex chemical composition, mainly represented by organic aromatic compounds. Wastewater pollutants significantly exceed discharge standards permissible for discharge into the Centralized Wastewater Disposal System of Settlements and Urban Districts. Phenol, which has the third hazard class according to the standards of maximum permissible concentrations of harmful substances in the waters of water bodies of fishery importance, is particularly dangerous. The toxic effect of phenol leads to the suppression of biocenoses, a decrease in the number of living organisms, negatively affects human health, and can lead to death. Based on the above, in order to reduce the anthropogenic load on the environment and maintain the quality of water resources, it is necessary to carry out a set of environmental measures aimed at wastewater treatment.The article presents methods of wastewater purification from phenol, describes complex purification methods and formulates proposals for the technology of wastewater treatment of the enterprise. Spectral and quantitative chemical analysis were performed to determine the composition of wastewater. Regenerative cleaning methods are proposed. Methods for wastewater treatment have been formulated and implemented. Methods for wastewater treatment of the coal tar processing process are proposed.The aims of research are to reduce the concentration of phenol in wastewater to the standard values permissible for discharge into the Centralized Wastewater Disposal System of Settlements and Urban Districts, based on experimental data; formulate a proposal for the wastewater treatment of the coal tar processing process from phenol.Objectives of the work are to study the composition of the source wastewater; to consider methods of wastewater treatment from phenols; to carry out wastewater treatment, based on the results of the study, to propose a wastewater treatment scheme.The object of the study is the wastewater of the enterprise after the coal tar processing process.

FAST, a method based on split-GFP for the detection in solution of proteins synthesized in cell-free expression systems

Cell-free protein synthesis (CFPS) systems offer a versatile platform for a wide range of applications. However, the traditional methods for detecting proteins synthesized in CFPS, such as radioactive labeling, fluorescent tagging, or electrophoretic separation, may be impractical, due to environmental hazards, high costs, technical complexity, and time consuming procedures. These limitations underscore the need for new approaches that streamline the detection process, facilitating broader application of CFPS. By harnessing the reassembly capabilities of two GFP fragments—specifically, the GFP1-10 and GFP11 fragments—we have crafted a method that simplifies the detection of in vitro synthesized proteins called FAST (Fluorescent Assembly of Split-GFP for Translation Tests). FAST relies on the fusion of the small tag GFP11 to virtually any gene to be expressed in CFPS. The in vitro synthesized protein:GFP11 can be rapidly detected in solution upon interaction with an enhanced GFP1-10 fused to the Maltose Binding Protein (MBP:GFP1-10). This interaction produces a fluorescent signal detectable with standard fluorescence readers, thereby indicating successful protein synthesis. Furthermore, if required, detection can be coupled with the purification of the fluorescent complex using standardized MBP affinity chromatography. The method's versatility was demonstrated by fusing GFP11 to four distinct E. coli genes and analyzing the resulting protein synthesis in both a homemade and a commercial E. coli CFPS system. Our experiments confirmed that the FAST method offers a direct correlation between the fluorescent signal and the amount of synthesized protein:GFP11 fusion, achieving a sensitivity threshold of 8 ± 2 pmol of polypeptide, with fluorescence plateauing after 4 h. Additionally, FAST enables the investigation of translation inhibition by antibiotics in a dose-dependent manner. In conclusion, FAST is a new method that permits the rapid, efficient, and non-hazardous detection of protein synthesized within CFPS systems and, at the same time, the purification of the target protein.

Design of cross-linkable polypept(o)ide-based AB3 miktoarm star polymers through advances in PeptoMiktoStar synthesis

Advances in polymer chemistry can provide convenient access to more complex polymeric architectures, such as miktoarm star polymers. In comparison to their linear counterparts, the physical properties of miktoarm star polymers can substantially differ. Their complex synthesis, characterization, and purification, however, makes it a challenging task to prepare well-defined structures with multiple arms of adjustable chemical nature. In this work, well-defined cross-linkable polypept(o)ide-based AB3 miktoarm star polymers were achieved, taking advantage of a novel asymmetric tetrafunctional initiator system with different orthogonal protecting groups. The first controlled living ring-opening polymerization (ROP) of sarcosine N-carboxyanhydride yields a three-armed polysarcosine (pSar) macroinitiator with low molecular weight dispersity (Đ ∼ 1.1), while a reactive polypeptide arm can be synthesized upon cleavage of the remaining protective group. The established reactive S-alkylsulfonyl protecting group bearing polypeptides poly(S-ethylsulfonyl-l-cysteine) (pCys(SO2Et)) and poly(S-ethylsulfonyl-l-homocysteine) (pHcy(SO2Et) were implemented into the asymmetric star architecture. The presence of the well-defined cross-linkable AB3 PeptoMiktoStars was verified through the characterization via 1H NMR, 1H DOSY, and SEC, revealing structures with dispersity indices of 1.1–1.2 and precise control over the degree of polymerization of each individual polymer chain. In aqueous solution, both types of PeptoMiktoStars form polymeric micelles. The size and morphology can be tuned by secondary structure directed self-assembly: the pHcy(SO2Et)20(pSar100)3 polymers form spherical micelles whilst the pCys(SO2Et)20(pSar100)3 assembled into worm-like micelles. Therefore, this synthetic methodology provides not only access to well-defined reactive miktoarm star polymers, but also allows for the engineering of polymeric micelles based on PeptoMiktoStars.

Lipopolysaccharide Regulates the Macrophage RNA-Binding Proteome.

RNA-protein interactions within cellular signaling pathways have significant modulatory effects on RNA binding proteins' (RBPs') effector functions. During the innate immune response, specific RNA-protein interactions have been reported as a regulatory layer of post-transcriptional control. We investigated changes in the RNA-bound proteome of immortalized mouse macrophages (IMM) following treatment with lipopolysaccharide (LPS). Stable isotope labeling by amino acids in cell culture (SILAC) of cells followed by unbiased purification of RNP complexes at two time points after LPS stimulation and bottom-up proteomic analysis by LC-MS/MS resulted in a set of significantly affected RBPs. Global RNA sequencing and LFQ proteomics were used to characterize the correlation of transcript and protein abundance changes in response to LPS at different time points with changes in protein-RNA binding. Il1α, MARCKS, and ACOD1 were noted as RBP candidates involved in innate immune signaling. The binding sites of the RBP and RNA conjugates at amino acid resolution were investigated by digesting the cross-linked oligonucleotide from peptides remaining after elution using Nuclease P1. The combined data sets provide directions for further studies of innate immune signaling regulation by RBP interactions with different classes of RNA.

Synergistic control potential of flue gas pollutants under Ultra-Low emission standards in waste incineration plants

As the dominant waste disposal process, incineration is regarded as the main incentive for the “not-in-my-backyard” syndrome, and faces an inescapable pressures of ultra-low emissions (ULE). Establishing precise response relationships between emission factors (EFs) and full-process influencing factors can provide guidance for the synergistic mitigation of flue gas pollutants (FGPs). In this work, the multi-dimensional EFs of FGPs were identified by initially integrating FGPs concentration monitoring data of existing 1,226 processing lines in China, technologies applied and operational experience (OE), local economic and political characteristics. Significant regional imbalance performance was observed, which EFs in the coastal regions were 3.55–92.39 % lower than those of the inland areas. NOx, SO2, HCl were identified as critical components requiring further reduction under the ULE standards, with exceedance rates recorded at 73.07 %, 38.90 %, and 56.69 %, respectively. An indicative value of 20 mg/m3 for PM is recommended for the control of heavy metals of Cd + Tl and Sb + As + Pb + Cr + Co + Cu + Mn + Ni based on the correlation coefficients of r = 0.28 (p < 0.001) and r = 0.20 (p = 0.002), respectively. Waste composition and OE were quantified as the main contributors of EFs’ disparities by the tree-branching controlled variable approach established in this study. Predictive models for FGPs control process and corresponding EFs were constructed. EFs of nine FGPs in 2030 would decrease by 0.97–65.42 %, due to more complex purification processes employed to meet ULE’s limitations, such as the application of five-stage processes growing from 45.60 % to 58.28 %. While regional imbalance in EFs-SO2 and EFs-HCl were extended with increases from 25.83 % to 33.07 % and 9.91 % to 32.32 %, respectively, due to the consistent disparities of OE and growing heterogeneity of control policies. Enhancing interregional empirical exchanges, reducing the regional market monopolies, and formulating technical guidelines would be beneficial to synergize the reduction of FGPs emissions and alleviate regional imbalance.