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Tao Cheng,1,2,&ast; Tian-Yong Han,1,2,&ast; Bo-Fu Liu,1,2 Pan Pan,1,2 Qiang Lai,1,2 Haifang Yu,1,2 Yu Cao1,2 1Emergency Department, West China Hospital of Sichuan University, Chengdu, Sichuan, People’s Republic of China; 2Laboratory of Emergency Medicine, West China Hospital of Sichuan University, Chengdu, Sichuan, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yu CaoEmergency Department of West China Hospital and Laboratory of Emergency Medicine of West China Hospital, Sichuan University, 37 Guoxue Road, Chengdu, 610041, Sichuan, People’s Republic of China, Tel +86-28-85422288, Email caoyu@wchscu.cnObjective: To develop a new approach based on Balthazar grades of acute pancreatitis (AP) and to assess this modified method as a tool for the early prediction of AP severity in the emergency department (ED).Methods: Data pertaining to AP patients ≥ 18 years old that had undergone computed tomography (CT) scanning within 24 h following ED admission between January 1, 2017 and September 30, 2017 were retrospectively analyzed. Patients were separated into two groups based on the length of time between the onset of their AP symptoms and the completion of CT scanning (Group 1: < 72 h; Group 2: ≥ 72 h). Modified Balthazar grades for these patients were then assessed, with the concordance between these modified grades and the 2012 revised Atlanta classification being assessed using the Kappa (κ) statistic. The modified grade with the largest κ value was evaluated based on performance traits including Harrell’s concordance index (C-index), area under the receiver operating characteristic curve (AUC) analyses, calibration curves, and decision curve analyses (DCA) in comparison with bedside index for severity in AP (BISAP) scores.Results: In total, 372 patients were included in the present analysis. These patients were regraded according to six methods, with the method yielding the largest κ value consisting of regraded Balthazar grades A–C, D, and E, respectively, corresponding to mild, moderate, and severe AP. The κ values for this method were 0.786 (95% CI, 0.706– 0.853) in Group 1 and 0.907 (95% CI, 0.842– 0.955) in Group 2, exhibiting nearly complete agreement with the latest Atlanta classification of AP. AUROC values for these modified Balthazar grades when used to predict SAP were significantly higher than those for BISAP scores in Group 1, Group 2, and the overall cohort (P < 0.05). The DCA curves for Group 1, Group 2, and the overall patient cohort exhibited substantial net benefits when using these modified grades across a range of POFs relative to BISAP scores. The calibration curve for this modified approach to predicting POF in AP patients revealed good agreement in this cohort.Conclusion: Modified Balthazar grades exhibited substantial to near-total agreement with the 2012 revised Atlanta classification of AP patients, and this modified method can thus be used for the early prediction of AP severity in the ED.Keywords: acute pancreatitis, severity, Balthazar grade, revised Atlanta classification

Heparin, a widely studied glycosaminoglycan, plays crucial roles in the regulation of various physiological and pathological processes. Therefore, it's important to develop highly selective and sensitive methods for convenient monitoring of heparin levels in biological systems. We report the design and synthesis of Fe 3 O 4 @PDA@MnO 2 nanoparticles (FPM-NPs), which exhibit dual enzymatic activities, enabling quantitative detection of heparin. The FPM-NPs feature a unique tri-layer spherical shell structure, possessing both peroxidase-like and oxidase-like activities, and catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence or absence of H 2 O 2 . Remarkably, upon co-incubated with heparin, the oxidase activity of FPM-NPs decreases, while the peroxidase activity increases. By leveraging these dual enzymatic properties of FPM-NPs, a highly sensitive and specific colorimetric detection of heparin is achieved, with a detection limit reaching 6.51 nM and a good linear response to quantify heparin ranging 10-800 nM. Additionally, the developed FPM-NPs are successfully applied to measure heparin in fetal bovine serum samples. We also extend this detection method to a paper-based chip, enabling portable detection of heparin through grayscale analysis of mobile phone photographs. The multi-nanozyme-based heparin detection approach provides a new perspective for future research on expanding the application of nanocomposite materials in biomedical detection and analysis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Microcolumn gel immunoassay (MGIA) has the ability to meet the requirements of clinical diagnosis due to its reliable sensitivity and accuracy. However, traditional MGIA exhibits limitations including inadequate portability, low throughput, and extended analysis time. To address these challenges, we combined MGIA with microfluidic technology, demonstrating a centrifugal microfluidic-based microcolumn gel immunoassay (μMGIA) platform for blood typing of clinical samples. Experimental results indicate that the μMGIA platform can simultaneously detect six blood group antigens in five clinical blood samples within 2 min. Notably, it offers comprehensive detection of ABO blood group antigens and Rh blood group antigens with 100 % accuracy, outperforming the traditional slide method. The integration of microfluidic technology with MGIA circumvents the constraints of traditional methods, providing a new avenue for blood typing and immunoanalysis of clinical samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Lignocellulosic biomass (LCB) is expected to play a critical role in achieving the goal of biomass-to-bioenergy conversion because of its wide distribution and low price. Biomass fermentation is a promising method for the sustainable generation of biohydrogen (bioH 2 ) from the renewable feedstock. Due to the inherent resistant structure of biomass, LCB needs to be pretreated to improve its digestibility and utilization. However, certain intermediates by-products generated during the pretreatment process, such as phenolic compounds, furan derivatives, and aldehydes, have been identified as potent inhibitors of subsequent anaerobic fermentation due to their disruptive effects on the physiological and metabolic functions of hydrogen-producing microbiota. To counteract the negative effects of these inhibitors on bio-H 2 fermentation, various detoxification strategies for LCB hydrolysates have been explored. This review presents a comprehensive analysis of fermentation-inhibitory by-products commonly generated by modern pretreatment protocols and their negative impacts on biohydrogen fermentation. Furthermore, the underlying mechanisms of inhibition upon hydrogen-producing microbes and their impacts on microbial community dynamics are exhibited. State-of-the-art strategies for detoxifying pretreated LCB have been also discussed, along with alternative pretreatment strategies designed to minimize or eliminate the formation of inhibitory by-products. Additionally, this review addresses the significant gap in the economic viability assessments of these processes, offering a detailed evaluation of both the technological and economic feasibility of biomass fermentation. Given the limitations of previous studies, strategies for cost-effective pretreatment and detoxification should be developed in the future to overcome the inhibition of fermentation inhibitors in the bioconversion of biomass to hydrogen., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Background: Simple, fast and low-cost paper-based analytical devices (PADs) have a good application prospect for point-of-care detection of GSH. However, effective immobilization of functional nanomaterials onto cellulose, as a critical factor in the construction of PADs, presents numerous difficulties and challenges., Results: In this study, we have developed an exceptionally straightforward and environmentally friendly synthetic approach by using ovalbumin (OVA) as a bio-mineralization template for the preparation of MnO 2 nanosheets. The MnO 2 nanosheets produced in the solution phase exhibited excellent intrinsic nano-enzyme activity and biodegradability. The OVA-MnO 2 nanosheets can effectively oxidize Amplex red in the absence of H 2 O 2 , enabling sensitive detection of GSH with a linear range of 5 nM-10 μM and a detection limit as low as 2.8 nM. Furthermore, we utilized this method to facilitate in situ synthesis of OVA-MnO 2 nanosheets directly on paper substrates. This approach eliminates the need for conventional stirring and centrifugation steps, greatly simplifying the fabrication process while reducing material usage and time expenditure. Characterization of the chemical composition and morphology confirmed the intimate growth of the 2D nano-enzymes on the cellulose fibers. Utilizing smartphone capabilities, the OVA-MnO 2 nanosheet-modified PAD enabled instrument-free detection of GSH, demonstrating high sensitivity (0.74 μM) and a wide linear response range (1-1000 μM)., Significance: The synthesis of MnO 2 nanosheets directly on cellulose substrates substantially streamlines the modification workflow of PADs and reduces detection costs, offering new avenues for clinical diagnostics of relevant diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Current researches on pesticides in wetlands are limited in terms of screening and quantification of many types of pesticides. Understanding the spatial and temporal dynamics, distribution patterns, and environmental risks of pesticides in multiple media is important for wetland ecological conservation. In this study, 222 pesticides were determined in multimedia samples collected simultaneously from the Songhua Wetland during four seasons. Concentrations of target pesticides in water, ice, sediment and soil ranged from 94.1 to 7445 ng/L, 62.6-953 ng/L, 0.82-50.2 ng/g dw, and 4.32-146 ng/g dw. Large spatial differences (p < 0.05) in pesticide concentrations in ice were found. However, there were no significant differences in the spatial and temporal distribution of pesticides in water, sediment, and soil (p > 0.05), suggesting that there were no correlation between the spatial and temporal use of pesticides. The dynamic exchange of pesticides between water-ice indicated that most pesticides were more enriched in water. However, there were still some pesticides (Dichlorvos and Biphenyl) that showed a stronger tendency to transfer from water to ice. Sediment-water exchange suggested that sediment is a source of secondary releases of most pesticides in wetland ecology, but is a sink for Biphenyl and Oxadiazon. The correlation between concentration ratios and fugacity fraction supported this finding. Most individual pesticides in wetland water and ice had shown low or moderate ecological risk conducted using risk quotient. The cumulative toxic effects of multiple pesticides had a high potential to pose a threat to wetland aquatic organisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Extracellular vesicles (EVs) play a crucial role in the occurrence and progression of cancer. The efficient isolation and analysis of EVs for early cancer diagnosis and prognosis have gained significant attention. In this study, for the first time, a rapid and visually detectable method termed freeze-thaw-induced floating patterns of gold nanoparticles (FTFPA) is proposed, which surpasses current state-of-the-art technologies by achieving a 100 fold improvement in the limit of detection of EVs. Notably, it allows for multi-dimensional visualizations of EVs through site-specific oligonucleotide incorporation. This capability empowers FTFPA to accurately identify EVs derived from subtypes of breast cancers with artificial intelligence algorithms. Intriguingly, learning the freezing-thawing-microstructures of EVs with a random forest algorithm is not only able to distinguish their original cell lines (with an accuracy of 95.56%), but also succeed in processing clinical samples (n = 156) to identify EVs by their healthy donors, breast lump and breast cancer subtypes (Luminal A, Triple-negative breast cancer, and Luminal B) with an accuracy of 83.33%. Therefore, this AI-empowered micro-visualization method establishes a rapid and precise point-of-care platform that is applicable to both fundamental research and clinical settings., (© 2024 Wiley‐VCH GmbH.)

Heavy metal pollution in the cold region is serious, affecting human health and aquatic ecology. This study investigated the ability of microalgae to remove heavy metals (HMs) and produce lipid at low temperature. The removal efficiency of different HMs (Cd 2+ , Cu 2+ , Cr 3+ and Pb 2+ ), cell growth and lipid synthesis of microalgae were analyzed at 15 °C. Moreover, addition of glycine betaine (GB) further enhanced the productivity of microalgae in treating HMs and lipid production, and simultaneously increased the antioxidant capacity of microalgae against environmental stresses. The results showed that the highest lipid productivity of 100.98 mg L -1 d -1 and the removal efficiency of 85.8 % were obtained under GB coupled with Cr 3+ . The highest glutathione content of 670.34 nmol g -1 fresh alga was achieved under GB coupled with Pb 2+ . In addition, lipidomics showed that GB was able to up-regulate the triglyceride and diglyceride content, influenced fatty acid composition to regulate the microalgal metabolism, and mediated lipid accumulation under 15 °C mainly through the regulation of glycerol ester metabolism. This study provided a new perspective on microalgal lipid production and the removal of HMs in cold regions and provided evidence for the use of phytohormones to improve the algal environmental resistance., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Anaerobic microorganisms are critical in regulating ethane in geothermal environments, where selenate and selenite are common contaminants. Although coupling ethane oxidation with selenate reduction has been demonstrated as feasible, such processes remain poorly explored in geothermal environments. This study addressed this gap by successfully enriching thermophilic anaerobic cultures capable of coupling ethane oxidation with selenate/selenite reduction, achieving selenate and selenite removal rate of 2.7 mg Se/L/d and 2.1 mg Se/L/d, respectively. Metagenomic analysis revealed a novel genus 'Candidatus Ethanivorans selenatireducens', which accounted for 16.0 % and 32.6 % of microbial communities in selenate- and selenite-dependent systems, respectively. This microorganism encoded pathways for anaerobic ethane oxidation via fumarate addition and genes required for the sequential reduction of selenate to elemental selenium. These findings unveiled a novel microbial mechanism linking ethane oxidation and selenate reduction in geothermal systems, providing new insights into the biogeochemical interaction between carbon and selenium in thermophilic environments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The aim of this study was to investigate the effects of various concentrations of antioxidants, including butyl hydroxy anisd (BHA), butylated hydroxytoluene (BHT), fulvic acid (FA), melatonin (MT), glycine betaine (GB) and putrescine (Put), on growth and lipid synthesis of microalgae under low-temperature (15 ℃). Changes in biochemical indicators, reactive oxygen species (ROS) level, glutathione (GSH) content and antioxidant enzyme activities were also studied. The results indicated that the maximum biomass concentration (1.3 g/L) and lipid productivity (75.3 ± 5.8 mg/L d -1 ) were achieved under 100 μM MT and 1 μM GB, respectively. Moreover, antioxidants were able to increase the GSH and antioxidant enzymes activities in algal cells under low-temperature stress. This study was enlightening for the utilization of antioxidants to improve the resistance to low-temperature stress and lipid production in microalgae, and provided a theoretical basis for the application of microalgae for lipid accumulation in cold regions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In microbiological research, traditional methods for bacterial screening and antibiotic susceptibility testing are resource-intensive. Microfluidics offers an efficient alternative with rapid results and minimal sample consumption, but the demand for cost-effective, user-friendly platforms persists in communities and hospitals. Inspired by the Magdeburg hemispheres, the strategy adapts to local conditions, leveraging omnipresent atmospheric pressure for self-sealing of Rotation-SlipChip (RSC) equipped with a 3D circular Christmas tree-like microfluidic concentration gradient generator. This innovative approach provides an accessible and adaptable platform for microbiological research and testing in diverse settings. The RSC can avoid leakage concerns during multiple concentration gradient generation, chip-rotating, and final long-term incubation reaction (≥24 h). Furtherly, RSC subtypes adapted to different reactions can be fabricated in less than 15 min with cost less than $1, the result can be read through designated observational windows by naked-eye. Moreover, the RSC demonstrates its capability for evaluating bacterial biomarker activity, enabling the rapid assessment of β-galactosidase concentration and enzyme activity within 30 min, and the limit of detection can be reduced by 10-fold. It also rapidly determines the minimum antibiotic inhibitory concentration and antibiotic combined medications results within 4 h. Overall, these low-cost and user-friendly RSC make them invaluable tools in determinations at previously impractical environment., (© 2024 Wiley‐VCH GmbH.)

Anaerobic ammonium oxidization (Anammox) process plays a crucial role in the global nitrogen cycle and sustainable biological nitrogen removal from wastewater. Although Anammox bacteria have been detected across mesophilic and thermophilic conditions, the direct cultivation of Anammox bacteria from thermal environments has remained elusive. This impedes limiting our understanding of their physiology and ecology in high-temperature habitats. Here, we successfully enriched Anammox bacteria from hot spring sediments at 45 °C, achieving an ammonium oxidation rate of 158.0 mg NH 4 + -N l -1 d -1 , with the genus 'Candidatus Brocadia' presenting 22.9 % of the total microbial community after about 500 days of operation. Metagenomic analysis recovered two high-quality genomes of novel Anammox bacteria, which we designed as 'Candidatus Brocadia thermophilus' and 'Candidatus Brocadia thermoanammoxidans'. Both of them encoded and actively expressed key metabolic genes involved in Anammox process and several genes associated with thermotolerance, demonstrating their remarkable ability to perform Anammox reaction in thermophilic environments. Notably, phylotypes related to 'Candidatus Brocadia thermoanammoxidans' have frequently been retrieved from geographically distinct natural habitats. These findings expand our understanding of thermophilic Anammox bacteria and underscore their potential in the nitrogen cycle of thermal natural and engineering ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In the intricately defined spatial microenvironment, a single fertilized egg remarkably develops into a conserved and well-organized multicellular organism. This observation leads us to hypothesize that stem cells or other seed cell types have the potential to construct fully structured and functional tissues or organs, provided the spatial cues are appropriately configured. Current organoid technology, however, largely depends on spontaneous growth and self-organization, lacking systematic guided intervention. As a result, the structures replicated in vitro often emerge in a disordered and sparse manner during growth phases. Although existing organoids have made significant contributions in many aspects, such as advancing our understanding of development and pathogenesis, aiding personalized drug selection, as well as expediting drug development, their potential in creating large-scale implantable tissue or organ constructs, and constructing multicomponent microphysiological systems, together with functioning at metabolic levels remains underutilized. Recent discoveries have demonstrated that the spatial definition of growth factors not only induces directional growth and migration of organoids but also leads to the formation of assembloids with multiple regional identities. This opens new avenues for the innovative engineering of higher-order organoids. Concurrently, the spatial organization of other microenvironmental cues, such as physical stresses, mechanical loads, and material composition, has been minimally explored. This review delves into the burgeoning field of organoid engineering with a focus on potential spatial microenvironmental control. It offers insight into the molecular principles, expected outcomes, and potential applications, envisioning a future perspective in this domain., Competing Interests: The authors have no conflicts to disclose., (© 2024 Author(s).)

For every epidemic outbreak, the prevention and treatments in resource-limited areas are always out of reach. Critical to this is that high accuracy, stability, and more comprehensive analytical techniques always rely on expensive and bulky instruments and large laboratories. Here, a fully integrated and high-throughput microfluidic system is proposed for ultra-multiple point-of-care immunoassay, termed Dac system. Specifically, the Dac system only requires a handheld portable device to automatically recycle repetitive multi-step reactions including on-demand liquid releasing, dispensing, metering, collecting, oscillatory mixing, and discharging. The Dac system performs high-precision enzyme-linked immunosorbent assays for up to 17 samples or targets simultaneously on a single chip. Furthermore, reagent consumption is only 2% compared to conventional ELISA, and microbubble-accelerated reactions shorten the assay time by more than half. As a proof of concept, the multiplexed detections are achieved by detecting at least four infection targets for two samples simultaneously on a singular chip. Furthermore, the barcode-based multi-target results can rapidly distinguish between five similar cases, allowing for accurate therapeutic interventions. Compared to bulky clinical instruments, the accuracy of clinical inflammation classification is 92.38% (n = 105), with a quantitative correlation coefficient of R 2 = 0.9838, while the clinical specificity is 100% and the sensitivity is 98.93%., (© 2024 Wiley‐VCH GmbH.)

Discs and numerous other consumer products have been developed for point of care testing (POCT) to replace traditional large and expensive biochemical devices in certain scenarios. Herein, we propose a drip-dry strategy (2D strategy) assisted Blu-ray disc (BD) biosensor, termed BDB, for rapid and portable POCT within 30 min with the cost of a single test < $1. The platform utilizes the covered area formed by the deposition of the substance to be measured on the activated BD surface after the evaporation of water and realizes the quantitative detection of the target through the error readout of free disc quality diagnosis software. As a proof of concept, we first demonstrated the feasibility of direct quantitative detection of substances in solution in a single system through the detection of pure proteins avoiding colorimetric reagent used in traditional optical detection. For the complex mixed systems, we then innovatively utilize the principle that soluble targets promote/inhibit the dissolution of insoluble precipitates to achieve specific detection of targets and successfully apply BDB to the indirect quantitative detection of glutathione (GSH) with LOD of 0.447 mM in the range of 2-16 mM and organophosphorus pesticides (OPs) with LOD of 2.122 × 10 -7 M in the range of 1.289 × 10 -7 -1.289 × 10 -4 M. The BDB is widely applicable, easy to operate, and less time-consuming, which is anticipated to provide an alternative method for early, on-site detection or screening.

Nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) process is a promising wastewater treatment technology, but the slow microbial growth rate greatly hinders its practical application. Although high-level nitrogen removal and excellent biomass accumulation have been achieved in n-DAMO granule process, the formation mechanism of n-DAMO granules remains unresolved. To elucidate the role of functional microbes in granulation, this study attempted to cultivate granules dominated by n-DAMO microorganisms and granules coupling n-DAMO with anaerobic ammonium oxidation (Anammox). After long-term operation, dense granules were developed in the two systems where both n-DAMO archaea and n-DAMO bacteria were enriched, whereas granulation did not occur in the other system dominated by n-DAMO bacteria. Extracellular polymeric substances (EPS) measurement indicated the critical role of EPS production in the granulation of n-DAMO process. Metagenomic and metatranscriptomic analyses revealed that n-DAMO archaea and Anammox bacteria were active in EPS biosynthesis, while n-DAMO bacteria were inactive. Consequently, more EPS were produced in the systems containing n-DAMO archaea and Anammox bacteria, leading to the successful development of n-DAMO granules. Furthermore, EPS biosynthesis in n-DAMO systems is potentially regulated by acyl-homoserine lactones and c-di-GMP. These findings not only provide new insights into the mechanism of granule formation in n-DAMO systems, but also hint at potential strategies for management of the granule-based n-DAMO process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Medium-chain fatty acids (MCFAs) production from waste activated sludge (WAS) by chain extension (CE) is a promising technology. However, the effects and mechanisms of CE process on the fate of antibiotic resistance genes (ARGs) remain unclear. In this study, the results showed that the removal efficiency of ARGs was 81.15 % in CE process, suggesting its efficacy in reducing environmental risks. Further, the observed decrease in mobile genetic elements (MGEs) indicated that CE process restricted the horizontal gene transfer (HGT). Complementing this, the increase in soluble organic matters and extracellular 16 S rDNA confirmed that MCFAs production caused bacterial damage. Decreased intracellular ARGs and increased extracellular ARGs further revealed that MCFAs production impaired ARGs hosts, thereby limiting the vertical gene transfer (VGT) of ARGs. Shift of microbial community combined with co-occurrence network analysis demonstrated that functional bacteria without host potential for ARGs were enriched, but potential ARGs and MGEs hosts decreased, showing the role of functional bacterial phylogeny and selection pressure of MCFAs in reducing ARGs. Finally, partial least squares path model was used to systematic verify the mechanism of ARGs removal in CE process, which was attributed to the inhibition of ARGs transmission (HGT and VGT) and shift of microbial community., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Phase separation inside mammalian cells regulates the formation of the biomolecular condensates that are related to gene expression, signalling, development and disease. However, a large population of endogenous condensates and their candidate phase-separating proteins have yet to be discovered in a quantitative and high-throughput manner. Here we demonstrate that endogenously expressed biomolecular condensates can be identified across a cell's proteome by sorting proteins across varying oligomeric states. We employ volumetric compression to modulate the concentrations of intracellular proteins and the degree of crowdedness, which are physical regulators of cellular biomolecular condensates. The changes in degree of the partition of proteins into condensates or phase separation led to varying oligomeric states of the proteins, which can be detected by coupling density gradient ultracentrifugation and quantitative mass spectrometry. In total, we identified 1,518 endogenous condensate proteins, of which 538 have not been reported before. Furthermore, we demonstrate that our strategy can identify condensate proteins that respond to specific biological processes., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Sulfate-dependent ammonium oxidation (Sulfammox) is a critical process linking nitrogen and sulfur cycles. However, the metabolic pathway of microbes driven Sulfammox is still in suspense. The study demonstrated that ammonium was not consumed with sulfate as the sole electron acceptor during long-term enrichment, probably due to inhibition from sulfide accumulation, while ammonium was removed at ∼ 10 mg N/L/d with sulfate and nitrate as electron acceptors. Ammonium and sulfate were converted into nitrogen gas, sulfide, and elemental sulfur. Sulfammox was mainly performed by Candidatus Brocadia sapporoensis and Candidatus Brocadia fulgida, both of which encoded ammonium oxidation pathway and dissimilatory sulfate reduction pathway. Not sulfide-driven autotrophic denitrifiers but Candidatus Kuenenia stuttgartiensis converted nitrate to nitrite with sulfide. The results of this study reveal the specialized metabolism of Sulfammox bacteria (Candidatus Brocadia sapporoensis and Candidatus Brocadia fulgida) and provide insight into microbial relationships during the nitrogen and sulfur cycles., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Public health events caused by pathogens have imposed significant economic and societal burdens. However, conventional methods still face challenges including complex operations, the need for trained operators, and sophisticated instruments. Here, we proposed a fully integrated and automated centrifugal microfluidic chip, also termed IACMC, for point-of-care multiplexed molecular diagnostics by harnessing the advantages of active and passive valves. The IACMC incorporates multiple essential components including a pneumatic balance module for sequential release of multiple reagents, a pneumatic centrifugation-assisted module for on-demand solution release, an on-chip silicon membrane module for nucleic acid extraction, a Coriolis force-mediated fluid switching module, and an amplification module. Numerical simulation and visual validation were employed to iterate and optimize the chip's structure. Upon sample loading, the chip automatically executes the entire process of bacterial sample lysis, nucleic acid capture, elution quantification, and isothermal LAMP amplification. By optimizing crucial parameters including centrifugation speed, direction of rotation, and silicone membrane thickness, the chip achieves exceptional sensitivity (twenty-five Salmonella or forty Escherichia coli) and specificity in detecting Escherichia coli and Salmonella within 40 min. The development of IACMC will drive advancements in centrifugal microfluidics for point-of-care testing and holds potential for broader applications in precision medicine including high-throughput biochemical analysis immune diagnostics, and drug susceptibility testing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) process offers a promising solution for simultaneously achieving methane emissions reduction and efficient nitrogen removal in wastewater treatment. Although nitrogen removal at a practical rate has been achieved by n-DAMO biofilm process, the mechanisms of biofilm formation and nitrogen transformation remain to be elucidated. In this study, n-DAMO biofilms were successfully developed in the membrane aerated moving bed biofilm reactor (MAMBBR) and removed nitrate at a rate of 159 mg NO 3 - -N L -1 d -1 . The obvious increase in the content of extracellular polymeric substances (EPS) indicated that EPS production was important for biofilm development. n-DAMO microorganisms dominated the microbial community, and n-DAMO bacteria were the most abundant microorganisms. However, the expression of biosynthesis genes for proteins and polysaccharides encoded by n-DAMO archaea was significantly more active compared to other microorganisms, suggesting the central role of n-DAMO archaea in EPS production and biofilm formation. In addition to nitrate reduction, n-DAMO archaea were revealed to actively express dissimilatory nitrate reduction to ammonium and nitrogen fixation. The produced ammonium was putatively converted to dinitrogen gas through the joint function of n-DAMO archaea and n-DAMO bacteria. This study revealed the biofilm formation mechanism and nitrogen-transformation network in n-DAMO biofilm systems, shedding new light on promoting the application of n-DAMO process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Waterborne pathogens invariably present considerable threats to public health. The quorum sensing (QS) system is instrumental in coordinating bacterial growth and metabolisms. However, the responses and regulatory mechanisms of bacteria to various disinfection technologies through quorum sensing are still unclear. This study examines the inactivation effect of chlorination and ozonation on biofilms and planktonic cells of QS signaling-deficient mutants of Pseudomonas aeruginosa . Cell counting and viability assessment revealed that the combined disinfection of chlorine and ozone was the most effective for inactivating planktonic P. aeruginosa within 10 min of exposure. Additionally, microfluidic chip culture demonstrated that the secretion of quinolone signals escalated biofilms' disinfection resistance. Disinfection exposure significantly altered the gene expression of wild-type strains and QS signaling-deficient mutants. Moreover, the QS system triggered multilayered gene expression programs as a responsive protection to disinfectant exposure, including oxidative stress, ribosome synthesis, and the nutrient absorption of bacteria. These insights broaden our understanding of bacterial QS in response to disinfection, promising potential strategies toward efficient disinfection processes.

Ethane, the second most abundant gaseous hydrocarbon in vast anoxic environments, is an overlooked greenhouse gas. Microbial anaerobic oxidation of ethane can be driven by available electron acceptors such as sulfate and nitrate. However, despite nitrite being a more thermodynamically feasible electron acceptor than sulfate or nitrate, little is known about nitrite-driven anaerobic ethane oxidation. In this study, a microbial culture capable of nitrite-driven anaerobic ethane oxidation was enriched through the long-term operation of a nitrite-and-ethane-fed bioreactor. During continuous operation, the nitrite removal rate and the theoretical ethane oxidation rate remained stable at approximately 25.0 mg NO 2 - N L -1  d -1 and 11.48 mg C 2 H 6 L -1  d -1 , respectively. Batch tests demonstrated that ethane is essential for nitrite removal in this microbial culture. Metabolic function analysis revealed that a species affiliated with a novel genus within the family Rhodocyclaceae, designated as ' Candidatus Alkanivoras nitrosoreducens', may perform the nitrite-driven anaerobic ethane oxidation. In the proposed metabolic model, despite the absence of known genes for ethane conversion to ethyl-succinate and succinate-CoA ligase, ' Ca . A. nitrosoreducens' encodes a prospective fumarate addition pathway for anaerobic ethane oxidation and a complete denitrification pathway for nitrite reduction to nitrogen. These findings advance our understanding of nitrite-driven anaerobic ethane oxidation, highlighting the previously overlooked impact of anaerobic ethane oxidation in natural ecosystems., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Point of care testing (POCT) of nucleic acids holds significant importance in the realm of infectious disease prevention and control, as well as the advancement of personalized precision medicine. Nevertheless, conventional nucleic acid testing methods continue to face challenges such as prolonged detection times and dependence on extensive specialized equipment and personnel, rendering them unsuitable for point of care applications. Here, we proposed an innovative active centrifugal microfluidic system (ACMS) for automatic nucleic acid extraction, encompassing modules for active valve control and magnetic control. An on-chip centrifugal puncture valve (PV) was devised based on the elastic tolerance differences between silicone membranes and tinfoils to release pre-embedded liquid reagents on demand. Furthermore, we have utilized the returnable valve (RV) technology to accurately control the retention and release of liquids, leveraging the high elastic tolerance of the silicone membrane. By incorporating an online controllable magnetic valve, we have achieved controlled and rapid aggregation and dispersion of magnetic beads. The final chip encapsulates multiple reagents and magnetic beads necessary for nucleic acid extraction. Upon sample addition and loading into the instrument, automated on-chip sample loading and nucleic acid extraction, purification, and collection can be accomplished within 30 minutes, halving the overall operation time and even increasing the efficiency of pseudovirus extraction by three orders of magnitude. Consequently, real-time fluorescence quantitative PCR amplification has successfully detected multiple targets of the SARS-CoV-2 virus (with an impressive detection limit as low as 10 copies per μL), along with targeted sequencing analysis yielding a conformity rate of 99%.

'Candidatus Methanoperedens nitroreducens' is an archaeal methanotroph with global importance that links carbon and nitrogen cycles and great potential for sustainable operation of wastewater treatment. It has been reported to mediate the anaerobic oxidation of methane through a reverse methanogenesis pathway while reducing nitrate to nitrite. Here, we demonstrate that 'Ca. M. nitroreducens' reduces ferric iron forming ammonium (23.1 %) and nitrous oxide (N 2 O, 46.5 %) from nitrate. These results are supported with the upregulation of genes coding for proteins responsible for dissimilatory nitrate reduction to ammonium (nrfA), N 2 O formation (norV, cyt P460), and multiple multiheme c-type cytochromes for ferric iron reduction. Concomitantly, an increase in the N 2 O-reducing SJA-28 lineage and a decrease in the nitrite-reducing 'Candidatus Methylomirabilis oxyfera' are consistent with the changes in 'Ca. M. nitroreducens' end products. These findings demonstrate the highly flexible physiology of 'Ca. M. nitroreducens' in anaerobic ecosystems with diverse electron acceptor conditions, and further reveals its roles in linking methane oxidation to global biogeochemical cycles. 'Ca. M. nitroreducens' could significantly affect the bioavailability of nitrogen sources as well as the emission of greenhouse gas in natural ecosystems and wastewater treatment plants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Immunoassays serve as powerful diagnostic tools for early disease screening, process monitoring, and precision treatment. However, the current methods are limited by high costs, prolonged processing times (>2 h), and operational complexities that hinder their widespread application in point-of-care testing. Here, we propose a novel centrifugo- p neumatic r eciprocating flowing coupled with spatial c onfinement strategy, termed PRCM , for ultrafast multiplexed immunoassay of pathogens on a centrifugal m icrofluidic platform. Each chip consists of four replicated units; each unit allows simultaneous detection of three targets, thereby facilitating high-throughput parallel analysis of multiple targets. The PRCM platform enables sequential execution of critical steps such as solution mixing, reaction, and drainage by coordinating inherent parameters, including motor rotation speed, rotation direction, and acceleration/deceleration. By integrating centrifugal-mediated pneumatic reciprocating flow with spatial confinement strategies, we significantly reduce the duration of immune binding from 30 to 5 min, enabling completion of the entire testing process within 20 min. As proof of concept, we conducted a simultaneous comparative test on- and off-the-microfluidics using 12 negative and positive clinical samples. The outcomes yielded 100% accuracy in detecting the presence or absence of the SARS-CoV-2 virus, thus highlighting the potential of our PRCM system for multiplexed point-of-care immunoassays.

Point-of-care testing (POCT) is experiencing a groundbreaking transformation with microfluidic chips, which offer precise fluid control and manipulation at the microscale. Nevertheless, chip design or operation for existing platforms is rather cumbersome, with some even heavily depending on external drivers or devices, impeding their broader utilization. This study develops a unique programmable gravity self-driven microfluidic chip (PGSMC) capable of simultaneous multi-reagent sequential release, multi-target analysis, and multi-chip operation. All necessary reagents are introduced in a single step, and the process is initiated simply by flipping the PGSMC vertically, eliminating the need for additional steps or devices. Additionally, it demonstrates successful immunoassays in less than 60 min for antinuclear antibodies testing, compared to more than 120 min by traditional methods. Assessment using 25 clinically diagnosed cases showcases remarkable sensitivity (96%), specificity (100%), and accuracy (99%). These outcomes underscored its potential as a promising platform for POCT with high accuracy, speed, and reliability, highlighting its capability for automated fluid control., (© 2023 Wiley‐VCH GmbH.)

Chlorinated organic pollutants constitute a significant category of persistent organic pollutants due to their widespread presence in the environment, which is primarily attributed to the expansion of agricultural and industrial activities. These pollutants are characterized by their persistence, potent toxicity, and capability for long-range dispersion, emphasizing the importance of their eradication to mitigate environmental pollution. While conventional methods for removing chlorinated organic pollutants encompass advanced oxidation, catalytic oxidation, and bioremediation, the utilization of biochar has emerged as a prominent green and efficacious method in recent years. Here we review biochar's role in remediating typical chlorinated organics, including polychlorinated biphenyls (PCBs), triclosan (TCS), trichloroethene (TCE), tetrachloroethylene (PCE), organochlorine pesticides (OCPs), and chlorobenzenes (CBs). We focus on the impact of biochar material properties on the adsorption mechanisms of chlorinated organics. This review highlights the use of biochar as a sustainable and eco-friendly method for removing chlorinated organic pollutants, especially when combined with biological or chemical strategies. Biochar facilitates electron transfer efficiency between microorganisms, promoting the growth of dechlorinating bacteria and mitigating the toxicity of chlorinated organics through adsorption. Furthermore, biochar can activate processes such as advanced oxidation or nano zero-valent iron, generating free radicals to decompose chlorinated organic compounds. We observe a broader application of biochar and bioprocesses for treating chlorinated organic pollutants in soil, reducing environmental impacts. Conversely, for water-based pollutants, integrating biochar with chemical methods proved more effective, leading to superior purification results. This review contributes to the theoretical and practical application of biochar for removing environmental chlorinated organic pollutants., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Waste activated sludge (WAS) as one of the major pollutants with a significant annual production, has garnered significant attention regarding its treatment and utilization. If improperly discharged, it not only caused environmental pollution but also led to the wastage of valuable resources. In this study, the microalgae growth and lipid accumulation using waste activated sludge extracts (WASE) under different temperature conditions were investigated. The highest lipid content (59.13%) and lipid productivity (80.41 mg L -1 d -1 ) were obtained at cultivation temperatures of 10 and 25 °C, respectively. It was found that microalgae can effectively utilize TN/TP/NH 4 + -N and other nutrients of WASE. The highest utilization rates of TP, TN and NH 4 + -N were achieved at a cultivation temperature of 10 °C, reaching 84.97, 77.49 and 92.32%, respectively. The algal fatty acids had carbon chains predominantly ranging from C14 to C18, making them suitable for biodiesel production. Additionally, a comprehensive analysis of transcriptomics and metabolomics revealed up-regulation of genes associated with triglyceride assembly, the antioxidant system of algal cells, and cellular autophagy, as well as the accumulation of metabolites related to the tricarboxylic acid (TCA) cycle and lipids. This study offers novel insights into the microscopic mechanisms of microalgae culture using WASE and approaches for the resource utilization of sludge., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

The development of more efficient advanced oxidation systems for serving various advanced treatment of wastewater is quite necessary and urgent. In this study, a nano-zero valent iron/periodate (nZVI-BC/PI) advanced oxidation system has been constructed, achieving a rapid degradation of acetaminophen (ACT, 1 mg/L) within 1 min (100 % at pH = 11) at low temperature (5℃). This system shows a great degradation in a wide range of pH (1 ∼ 11), improving the pH limitation of PI oxidation system. During the reaction process, ·OH as the main active species collaborate with 1 O 2 , Fe (IV), ·O 2 - and electron transfer to degrade ACT. In this system, iron ion leaching is low (0.019 mg/L), ACT was effectively degraded (74.36 %∼97.32 %) under different water, moreover, the material has an expected recyclability. The research provides a significant guidance for the advanced treatment of wastewater especially in cold regions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A coupling technology for lipid production and adsorption of rare earth elements (REEs) using microalgae was studied in this work. The microalgae cell growth, lipid production, biochemical parameters and lipid profiles were investigated under different REEs (Ce 3+ , Gd 3+ and La 3+ ). The results showed that the maximum lipid production was achieved at different concentrations of REEs, with lipid productivities of 300.44, 386.84 and 292.19 mg L -1 d -1 under treatment conditions of 100 μg L -1 Ce 3+ , 250 μg L -1 Gd 3+ and 1 mg L -1 La 3+ , respectively. Moreover, the adsorption efficiency of Ce 3+ , Gd 3+ and La 3+ exceeded 96.58 %, 93.06 % and 91.3 % at concentrations of 25-1000 μg L -1 , 100-500 μg L -1 and 0.25-1 mg L -1 , respectively. In addition, algal cells were able to adsorb 66.2 % of 100 μg L -1 Ce 3+ , 48.4 % of 250 μg L -1 Gd 3+ and 59.9 % of 1 mg L -1 La 3+ . The combination of extracellular polysaccharide and algal cell wall could adsorb 25.2 % of 100 μg L -1 Ce 3+ , 44.5 % of 250 μg L -1 Gd 3+ and 30.5 % of 1 mg L -1 La 3+ , respectively. These findings indicated that microalgae predominantly adsorbed REEs through the intracellular pathway. This study elucidates the mechanism of effective lipid accumulation and adsorption of REEs by microalgae under REEs stress conditions. It establishes a theoretical foundation for the efficient microalgae lipid production and REEs recovery from wastewater or waste residues containing REEs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Due to the ever-increasing challenge of emerging and reemerging infections on global health, the development of POCT tools has been propelled. However, conventional point-of-care testing methods suffer from several limitations, including cumbersome operation, long detection times, and low accuracy, which hamper their widespread application. Compared to traditional disease diagnostic equipment, mobile health platforms offer several advantages, including portability, ease of operation, and automated analysis of detection results through recognition algorithms. Consequently, they hold great promise for the future. Here, we developed a smartphone-based centrifugal mHealth platform implementing daisy-shaped quick response chip for hematocrit measurement. The centrifugal microfluidic chip is combined with a smartphone through a back-clip-on mobile phone adapter whose control circuit is designed with low power consumption to enable the platform to operate without requiring a high-power source that is inconvenient to carry, thereby achieving the goal of portability. Concurrently, we designed a quick response chip featuring a unique hollow daisy structure that is in line with the properties of hematocrit detection. The distinctive configuration of the chip enables adequate centrifugal force to be supplied for hematocrit detection. Additionally, our customized quick response code recognition algorithm is able to recognize this chip, facilitating non-experts in performing hematocrit intelligent recognition with their smartphones., Competing Interests: Declaration of competing interest There are no conflicts of interest to declare., (Copyright © 2023. Published by Elsevier B.V.)

Exosomal microRNAs (miRNAs) play a pivotal role in intercellular communication, regulating gene expression in target cells, and hold significant promise as cancer biomarkers for early detection and screening. However, achieving precise and viable detection of exosomal miRNAs remains a challenge. This paper proposes an all-in-one detection strategy for breast cancer-derived exosomal miRNA-21 on a pen-based paper chip (PPC). The PPC is constructed using a modified automatic pen and lateral flow assay (LFA), which results in a cost-effective fabrication process. The user only needs to add the sample and trigger the top of the self-contained PPC after a period of time to complete the entire detection process. To enhance the sensitivity of exosomal miRNA testing, an enzyme-free catalyzed hairpin assembly (CHA) is further introduced, enabling highly sensitive detection of miRNA-21 with a limit of detection (LOD) of 25 fmol. Additionally, the detection of miRNAs in differentially-expressed cells and clinical samples has also been successfully achieved with high specificity. Overall, the proposed PPC provides an effective tool for detecting early cancer, monitoring diseases, and establishing point of care testing (POCT).

Background: Ovarian cancer (OC) is one of the most common gynecological tumors with high morbidity and mortality. Altered serum N-glycome has been observed in many diseases, while the association between serum protein N-glycosylation and OC progression remains unclear, particularly for the onset of carcinogenesis from benign neoplasms to cancer., Methods: Herein, a mass spectrometry based high-throughput technique was applied to characterize serum N-glycome profile in individuals with healthy controls, benign neoplasms and different stages of OC. To elucidate the alterations of glycan features in OC progression, an orthogonal strategy with lectin-based ELISA was performed., Results: It was observed that the initiation and development of OC was associated with increased high-mannosylationand agalactosylation, concurrently with decreased total sialylation of serum, each of which gained at least moderately accurate merits. The most important individual N-glycans in each glycan group was H7N2, H3N5 and H5N4S2F1, respectively. Notably, serum N-glycome could be used to accurately discriminate OC patients from benign cohorts, with a comparable or even higher diagnostic score compared to CA125 and HE4. Furthermore, bioinformatics analysis based discriminative model verified the diagnostic performance of serum N-glycome for OC in two independent sets., Conclusions: These findings demonstrated the great potential of serum N-glycome for OC diagnosis and precancerous lesion prediction, paving a new way for OC screening and monitoring., (© 2024. The Author(s).)

The abuse of antibiotics has become a global public safety issue, leading to the development of antimicrobial resistance (AMR). The development of antimicrobial susceptibility testing (AST) is crucial in reducing the growth of AMR. However, traditional AST methods are time-consuming (e.g., 24-72 h), labor-intensive, and costly. Here, we propose a controlled-diffusion centrifugal microfluidic platform (CCM) for rapid AST to obtain highly precise minimum inhibitory concentration (MIC) values. Antibiotic concentration gradients are generated by controlled moving and diffusing of antibiotic and buffer solution along the main microchannel within 3 min. The solution and bacterial suspension are then injected into the outermost reaction chamber by simple centrifugation. The CCM successfully determined the MIC for three commonly used antibiotics in clinical settings within 4-9 h. To further enhance practicality, reduce costs, and meet point-of-care testing demands, we have developed an integrated mobile detection platform for automated MIC value acquisition. The proposed CCM is a simple, low-cost, and portable method for rapid AST with broad clinical and in vitro applications., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Biomacromolecular folding kinetics involves fast folding events and broad timescales. Current techniques face limitations in either the required time resolution or the observation window. In this study, we developed the TeZla micromixer, integrating Tesla and Zigzag microstructures with a multistage velocity descending strategy. TeZla achieves a significant short mixing dead time (40 µs) and a wide time window covering four orders of magnitude (up to 300 ms). Using this unique micromixer, we explored the folding landscape of c-Myc G4 and its noncanonical-G4 derivatives with different loop lengths or G-vacancy sites. Our findings revealed that c-Myc can bypass folding intermediates and directly adopt a G4 structure in the cation-deficient buffer. Moreover, we found that the loop length and specific G-vacancy site could affect the folding pathway and significantly slow down the folding rates. These results were also cross-validated with real-time NMR and circular dichroism. In conclusion, TeZla represents a versatile tool for studying biomolecular folding kinetics, and our findings may ultimately contribute to the design of drugs targeting G4 structures., Competing Interests: Competing interests statement:The authors declare no competing interest.

Nitrate/nitrite-dependent anaerobic oxidation of methane (n-DAMO) is a recently discovered process, which provides a sustainable perspective for simultaneous nitrogen removal and greenhouse gas emission (GHG) mitigation by using methane as an electron donor for denitrification. However, the engineering roadmap of the n-DAMO process is still unclear. This work constitutes a state-of-the-art review on the classical and most recently discovered metabolic mechanisms of the n-DAMO process. The versatile combinations of the n-DAMO process with nitrification, nitritation, and partial nitritation for nitrogen removal are also clearly presented and discussed. Additionally, the recent advances in bioreactor development are systematically reviewed and evaluated comprehensively in terms of methane supply, biomass retention, membrane requirement, startup time, reactor performance, and limitations. The key issues including enrichment and operation strategy for the scaling up of n-DAMO-based processes are also critically addressed. Moreover, the challenges inherent to implementing the n-DAMO process in practical applications, including application scenario recognition, GHG emission mitigation, and operation under realistic conditions, are highlighted. Finally, prospects as well as opportunities for future research are proposed. Overall, this review provides a roadmap for potential applications and further development of the n-DAMO process in the field of wastewater treatment.

Wnt/β-catenin signaling is a conserved pathway crucially governing development, homeostasis, and oncogenesis. Discoveries of its regulators hold great values in both basic and translational research. Through screening, we identified a deubiquitinase, USP10, as a critical modulator of β-catenin. Mechanistically, USP10 binds to key scaffold Axin1 via conserved motifs and stabilizes Axin1 through K48-linked deubiquitination. Surprisingly, USP10 physically tethers Axin1 and β-catenin and promotes the phase separation for β-catenin suppression regardless of the enzymatic activity. Function-wise, USP10 enzymatic activity preferably regulates embryonic development and both the enzymatic activity and physical function jointly control intestinal homeostasis by antagonizing β-catenin. In colorectal cancer, USP10 substantially represses cancer growth mainly through physical promotion of phase separation and correlates with Wnt/β-catenin magnitude clinically. Collectively, we discovered USP10 functioning in multiple biological processes against β-catenin and unearthed the enzyme-dependent and -independent "dual-regulating" mechanism. These two functions of USP10 work in parallel and are context dependent., Competing Interests: Declaration of interests The authors declare no conflict of interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Objective: To investigate the expression of neuraminidase-1 (NEU1) in Ewing sarcoma (ES) tissue and its effect on the proliferation and migration of ES cells. Methods: To obtain datasets of ES from the National Center for Biotechnology Information's High-Throughput Gene Expression Omnibus (GEO) for the analysis of NEU1 expression in ES; to acquire ES patient dataset from the International Cancer Genome Consortium (ICGC) database and apply Kaplan-Meier survival analysis to investigate the relationship between NEU1 and the prognosis of ES patients; adopting both univariate and multivariate Cox regression analysis to determine whether NEU1 is a prognostic factor for ES; adopting the Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation to analyze the potential mechanism of NEU1 in regulating the malignant biological behavior of ES; adopting the real-time fluorescence quantitative polynucleotide chain reaction (RT-qPCR) to verify the expression of NEU1 in the human bone marrow mesenchymal stem cells (hBMSC) and the ES cell line RD-ES; adopting the transfection technology to knock down the expression of NEU1 in ES cell lines and divide them into two groups: shRNA-NEU1 and shRNA-NC to explore the effects of altered NEU1 expression on ES malignant behavior; adopting the cell counting kit (CCK-8) and cell clone formation experiment to detect the proliferation ability of two groups of cells; adopting the scratch healing experiment to test the cell migration ability of the two groups. Results: We retrieved and analyzed data from the GEO database, including GSE17674 (44 ES tissues and 18 normal tissues) and GSE17679 (87 ES tissues and 18 normal tissues), and found that NEU1 expression was significantly higher in ES tissues compared to normal control tissues ( P <0.001). The complete gene expression and clinical information of 56 ES patients obtained from the ICGC database revealed that the ES patients with high NEU1 expression ( n =28) had a significantly lower overall survival rates at different time points compared to those with low NEU1 expression ( n =28) ( HR =2.830, 95% CI :1.324-6.051, P =0.005). Univariate analysis indicated that NEU1 could impact ES patient prognosis ( HR =1.049, 95% CI : 1.008-1.092, P =0.019), and multivariate analysis further suggested that NEU1 could serve as a risk factor for ES prognosis ( HR =1.087, 95% CI : 1.028-1.148, P =0.003). KEGG results show that MAPK signaling pathway and cell adhesion molecule signaling pathway were potential mechanisms regulating the malignant process of ES. The RT-qPCR results showed that the expression level of NEU1 in the RD-ES cell line is significantly higher than that in the control cell hBMSC (2 184.23±527.32 vs 1.00±0.08, P <0.001). The CCK-8 experiment results show that the proliferation of RD-ES cells in the NEU1 knockdown group was lower than that in the control group at 24, 48, and 72 hours (0.494±0.126 vs 0.696±0.118, 0.657±0.096 vs 1.142±0.182, 1.053±0.064 vs 1.980±0.146, all P <0.001). The results of single cell clone formation experiment show that the number of colony formation in the low expression NEU1 group was significantly lower than that in the control group (184.2±123.9 vs 362.8±78.0, P =0.021). The cell scratch healing experiment finds that the average scratch distance of the NEU1 knockdown group was significantly lower than that of the control group (19.6%±5.7% vs 56.0%±7.6%, P <0.001). Conclusion: NEU1 may be a prognostic factor in ES, and its abnormal expression in ES can affect the proliferation and migration ability of the ES cells, leading to poor prognosis in ES patients.

This study proposes a new model in which ethanol and acetate produced by dark fermentation are processed by Clostridium kluyveri for chain elongation to produce caproate with an addition of biochar prepared from cornstalk residues after acid pretreatment and enzymatic hydrolysis (AERBC) in the dark fermentation and chain elongation processes. The results show a 6-25% increase in hydrogen production in dark fermentation with adding AERBC, and the maximum concentration of caproate in the new model reached 1740 mg/L, 61% higher than that in the control group. In addition, caproate was obtained by dark fermentation, using liquid metabolites as substrates with an initial pH range of 6.5-7.5. Finally, the electron balance and electron transfer efficiency in the new model were analyzed, and the role of AERBC in dark fermentation and chain elongation was investigated. This study provides a new reference for the use of dark-fermented liquid metabolites and cornstalk residue., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Thermophilic microorganisms mediated significant element cycles and material conversion in the early Earth as well as mediating current thermal environments. Over the past few years, versatile microbial communities that drive the nitrogen cycle have been identified in thermal environments. Understanding the microbial-mediated nitrogen cycling processes in these thermal environments has important implications for the cultivation and application of thermal environment microorganisms as well as for exploring the global nitrogen cycle. This work provides a comprehensive review of different thermophilic nitrogen-cycling microorganisms and processes, which are described in detail according to several categories, including nitrogen fixation, nitrification, denitrification, anaerobic ammonium oxidation, and dissimilatory nitrate reduction to ammonium. In particular, we assess the environmental significance and potential applications of thermophilic nitrogen-cycling microorganisms, and highlight knowledge gaps and future research opportunities., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Objective: To establish a systematic, scientific, reasonable, feasible and reliable evaluation system for occupational disease prevention and control capability, in order to provide reference for occupational disease prevention and control work in Tianjin City. Methods: In August 2022, literature review was conducted to propose indicators for the evaluation system. Two rounds of anonymous consultation with occupational health experts were conducted using the Delphi method to form expert opinions. According to the boundary value method and expert opinions, eliminate, screen, and modify the evaluation system indicators to ultimately determine the system indicators. Use Cronbach's alpha to test the reliability of the system indicators and form a Tianjin occupational disease prevention and control capability evaluation system. Results: It showed that the effective response rates of the two rounds of consultation conducted by experts in this study were 92.3% and 100.0%, respectively. The expert authority coefficients were 0.84 and 0.82, respectively. The Kendall coordination coefficient was tested for differences, and the differences were statistically significant ( P <0.05). The Tianjin occupational disease prevention and control capability evaluation system includes 7 primary indicators, 17 secondary indicators, and 54 tertiary indicators. The Cronbach's alpha of the primary, second, third level indicators and all indicators were 0.91, 0.98, 0.98, 0.98 (>0.7) . Conclusion: The preliminary evaluation system for occupational disease prevention and control capacity in Tianjin City has been established, providing a reference basis for the investigation of occupational disease prevention and control capacity in Tianjin City.

Granular sludge combined n-DAMO and Anammox (n-D/A) is an energy-efficient biotechnique for the simultaneous removal of nitrogen and dissolved methane from wastewater. However, the lack of knowledge so far about the metabolic interactions between n-DAMO and Anammox in response to operation condition in granular sludge restrains the development of this biotechnology. To address this gap, three independent membrane granular sludge reactors (MGSRs) were designed to carry out the granule-based n-D/A process under different conditions. We provided the first deep insights into the metabolic interactions between n-DAMO and Anammox in granular sludge via combined metagenomic and metatranscriptomic analyses. Our study unveiled a clear population shift of n-DAMO community from Candidatus Methanoperedens to Candidatus Methylomirabilis from sidestream to mainstream. Candidatus Methanoperedens with relative abundance of 25.2% played the major role in nitrate reduction and methane oxidation under sidestream condition, indicated by the high expression activities of mcrA and narG. Candidatus Methylomirabilis dominated the microbial community under mainstream condition with relative abundance of 32.1%, supported by the high expression activities of pmoA and hao. Furthermore, a transition of Anammox population from Candidatus Kuenenia to Candidatus Brocadia was also observed from sidestream to mainstream. Candidatus Kuenenia and Candidatus Brocadia jointly contributed to the primary anaerobic ammonium oxidation suggested by the high expression value of hdh and hzs. Candidatus Methylomirabilis was speculated to perform ammonium oxidation mediated by pMMO under mainstream condition. These findings might help to reveal the microbial interactions and ecological niches of n-DAMO and Anammox microorganisms, shedding light on the optimization and management of the granule-based n-D/A system., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Droplet microfluidics is a rapidly advancing area of microfluidic technology, which offers numerous advantages for cell analysis, such as isolation and accumulation of signals, by confining cells within droplets. However, controlling cell numbers in droplets is challenging due to the uncertainty of random encapsulation which result in many empty droplets. Therefore, more precise control techniques are needed to achieve efficient encapsulation of cells within droplets. Here, an innovative microfluidic droplet manipulation platform had been developed, which employed positive pressure as a stable and controllable driving force for manipulating fluid within chips. The air cylinder, electro-pneumatics proportional valve, and the microfluidic chip were connected through a capillary, which enabled the formation of a fluid wall by creating a difference in hydrodynamic resistance between two fluid streams at the channel junction. Lowering the pressure of the driving oil phase eliminates hydrodynamic resistance and breaks the fluid wall. Regulating the duration of the fluid wall breakage controls the volume of the introduced fluid. Several important droplet microfluidic manipulations were demonstrated on this microfluidic platform, such as sorting of cells/droplets, sorting of droplets co-encapsulated cells and hydrogels, and active generation of droplets encapsulated with cells in a responsive manner. The simple, on-demand microfluidic platform was featured with high stability, good controllability, and compatibility with other droplet microfluidic technologies., (© 2023. Springer-Verlag GmbH Germany, part of Springer Nature.)