Coxiella burnetii effector CvpE maintains biogenesis of Coxiella-containing vacuoles by suppressing lysosome tubulation through binding PI(3)P and perturbing PIKfyve activity on lysosomes.

Coxiella burnetii (C. burnetii) is the causative agent of Q fever, a zoonotic disease. Intracellular replication of C. burnetii requires the maturation of a phagolysosome-like compartment known as the replication permissive Coxiella-containing vacuole (CCV). Effector proteins secreted by the Dot/Icm secretion system are indispensable for maturation of a single large CCV by facilitating the fusion of promiscuous vesicles. However, the mechanisms of CCV maintenance and evasion of host cell clearance remain to be defined. Here, we show that C. burnetii secreted Coxiella vacuolar protein E (CvpE) contributes to CCV biogenesis by inducing lysosome-like vacuole (LLV) enlargement. LLV fission by tubulation and autolysosome degradation is impaired in CvpE-expressing cells. Subsequently, we found that CvpE suppresses lysosomal Ca2+ channel transient receptor potential channel mucolipin 1 (TRPML1) activity in an indirect manner, in which CvpE binds phosphatidylinositol 3-phosphate [PI(3)P] and perturbs PIKfyve activity in lysosomes. Finally, the agonist of TRPML1, ML-SA5, inhibits CCV biogenesis and C. burnetii replication. These results provide insight into the mechanisms of CCV maintenance by CvpE and suggest that the agonist of TRPML1 can be a novel potential treatment that does not rely on antibiotics for Q fever by enhancing Coxiella-containing vacuoles (CCVs) fission.

A universal and scalable transformation of bulk metals into single-atom catalysts in ionic liquids.

Single-atom catalysts (SACs) with maximized metal atom utilization and intriguing properties are of utmost importance for energy conversion and catalysis science. However, the lack of a straightforward and scalable synthesis strategy of SACs on diverse support materials remains the bottleneck for their large-scale industrial applications. Herein, we report a general approach to directly transform bulk metals into single atoms through the precise control of the electrodissolution-electrodeposition kinetics in ionic liquids and demonstrate the successful applicability of up to twenty different monometallic SACs and one multimetallic SAC with five distinct elements. As a case study, the atomically dispersed Pt was electrodeposited onto Ni3N/Ni-Co-graphene oxide heterostructures in varied scales (up to 5 cm × 5 cm) as bifunctional catalysts with the electronic metal-support interaction, which exhibits low overpotentials at 10 mA cm-2 for hydrogen evolution reaction (HER, 30 mV) and oxygen evolution reaction (OER, 263 mV) with a relatively low Pt loading (0.98 wt%). This work provides a simple and practical route for large-scale synthesis of various SACs with favorable catalytic properties on diversified supports using alternative ionic liquids and inspires the methodology on precise synthesis of multimetallic single-atom materials with tunable compositions.

TRIM56-mediated production of type I interferon inhibits intracellular replication of Rickettsia rickettsii.

Rickettsia rickettsii (R. rickettsii), the causative agent of Rocky Mountain spotted fever (RMSF), is the most pathogenic member among Rickettsia spp. Previous studies have shown that tripartite motif-containing 56 (TRIM56) E3 ligase-induced ubiquitination of STING is important for cytosolic DNA sensing and type I interferon production to induce anti-DNA viral immunity, but whether it affects intracellular replication of R. rickettsii remains uncharacterized. Here, we investigated the effect of TRIM56 on HeLa and THP-1 cells infected with R. rickettsii. We found that the expression of TRIM56 was upregulated in the R. rickettsii-infected cells, and the overexpression of TRIM56 inhibited the intracellular replication of R. rickettsii, while R. rickettsii replication was enhanced in the TRIM56-silenced host cells with the reduced phosphorylation of IRF3 and STING and the increased production of interferon-ß. In addition, the mutation of the TRIM56 E3 ligase catalytic site impairs the inhibitory function against R. rickettsii in HeLa cells. Altogether, our study discovers that TRIM56 is a host restriction factor of R. rickettsii by regulating the cGAS-STING-mediated signaling pathway. This study gives new evidence for the role of TRIM56 in the innate immune response against intracellular bacterial infection and provides new therapeutic targets for RMSF. IMPORTANCE: Given that Rickettsia rickettsii (R. rickettsii) is the most pathogenic member within the Rickettsia genus and serves as the causative agent of Rocky Mountain spotted fever, there is a growing need to explore host targets. In this study, we examined the impact of host TRIM56 on R. rickettsii infection in HeLa and THP-1 cells. We observed a significant upregulation of TRIM56 expression in R. rickettsii-infected cells. Remarkably, the overexpression of TRIM56 inhibited the intracellular replication of R. rickettsii, while silencing TRIM56 enhanced bacterial replication accompanied by reduced phosphorylation of IRF3 and STING, along with increased interferon-ß production. Notably, the mutation of the TRIM56's E3 ligase catalytic site did not impede R. rickettsii replication in HeLa cells. Collectively, our findings provide novel insights into the role of TRIM56 as a host restriction factor against R. rickettsii through the modulation of the cGAS-STING signaling pathway.

Sintering-induced cation displacement in protonic ceramics and way for its suppression.

Protonic ceramic fuel cells with high efficiency and low emissions exhibit high potential as next-generation sustainable energy systems. However, the practical proton conductivity of protonic ceramic electrolytes is still not satisfied due to poor membrane sintering. Here, we show that the dynamic displacement of Y3+ adversely affects the high-temperature membrane sintering of the benchmark protonic electrolyte BaZr0.1Ce0.7Y0.1Yb0.1O3-δ, reducing its conductivity and stability. By introducing a molten salt approach, pre-doping of Y3+ into A-site is realized at reduced synthesis temperature, thus suppressing its further displacement during high-temperature sintering, consequently enhancing the membrane densification and improving the conductivity and stability. The anode-supported single cell exhibits a power density of 663 mW cm-2 at 600 °C and long-term stability for over 2000 h with negligible performance degradation. This study sheds light on protonic membrane sintering while offering an alternative strategy for protonic ceramic fuel cells development.

Manganese-Based Nanoparticle Vaccine for Combating Fatal Bacterial Pneumonia.

Bacterial pneumonia is the leading cause of death worldwide among all infectious diseases. However, currently available vaccines against fatal bacterial lung infections, e.g., pneumonic plague, are accompanied by limitations, including insufficient antigen-adjuvant co-delivery and inadequate immune stimulation. Therefore, there is an urgent requirement to develop next-generation vaccines to improve the interaction between antigen and adjuvant, as well as enhance the effects of immune stimulation. This study develops a novel amino-decorated mesoporous manganese silicate nanoparticle (AMMSN) loaded with rF1-V10 (rF1-V10@AMMSN) to prevent pneumonic plague. These results suggest that subcutaneous immunization with rF1-V10@AMMSN in a prime-boost strategy induces robust production of rF1-V10-specific IgG antibodies with a geometric mean titer of 315,844 at day 42 post-primary immunization, which confers complete protection to mice against 50 × LD50 of Yersinia pestis (Y. pestis) challenge via the aerosolized intratracheal route. Mechanistically, rF1-V10@AMMSN can be taken up by dendritic cells (DCs) and promote DCs maturation through activation of the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway and production of type I interferon. This process results in enhanced antigen presentation and promotes rF1-V10-mediated protection against Y. pestis infection. This manganese-based nanoparticle vaccine represents a valuable strategy for combating fatal bacterial pneumonia.

Construction of an inhalable recombinant M2e-FP-expressing Bacillus subtilis spores-based vaccine and evaluation of its protection efficacy against influenza in a mouse model.

Influenza A virus (IAV) is a deadly zoonotic pathogen that remains a burden to global health systems despite continuous vaccinations, indicating the need for an improved vaccine strategy. In this work, we constructed a new recombinant influenza vaccine using Bacillus subtilis spores expressing M2e-FP protein (RSM2eFP) and assessed its potency and efficacy in BALB/c mouse immunized via aerosolized intratracheal inoculation (i.t.) or intragastric (i.g.) administration. Immunization via i.t. route conferred 100 % protection against 20 × LD50 A/PR/8/34 (H1N1) virus compared with only 50 % via the i.g. route. Even when challenged with 40 × LD50 virus, the RSM2eFP vaccine immunized via i.t. provided 80 % protection. Consistently, i.t. inoculation of RSM2eFP spore vaccine induced a stronger lung mucosal immune response and a greater cellular immune response than i.g. administration, as indicated by the high production of IgG and SIgA. In addition, the RSM2eFP spore vaccine diminished the yield of infectious virus in the lung of mice immunized via i.t. These results suggest that i.t. immunization of the RSM2eFP spore vaccine may be a promising strategy for the development of mucosal vaccines against IAV infections.

Molten salt electrosynthesis of Cr2GeC nanoparticles as anode materials for lithium-ion batteries.

The two-dimensional MAX phases with compositional diversity are promising functional materials for electrochemical energy storage. Herein, we report the facile preparation of the Cr2GeC MAX phase from oxides/C precursors by the molten salt electrolysis method at a moderate temperature of 700°C. The electrosynthesis mechanism has been systematically investigated, and the results show that the synthesis of the Cr2GeC MAX phase involves electro-separation and in situ alloying processes. The as-prepared Cr2GeC MAX phase with a typical layered structure shows the uniform morphology of nanoparticles. As a proof of concept, Cr2GeC nanoparticles are investigated as anode materials for lithium-ion batteries, which deliver a good capacity of 177.4 mAh g-1 at 0.2 C and excellent cycling performance. The lithium-storage mechanism of the Cr2GeC MAX phase has been discussed based on density functional theory (DFT) calculations. This study may provide important support and complement to the tailored electrosynthesis of MAX phases toward high-performance energy storage applications.

Ordered Assembly of DNA on Topological Insulator Bi2Se3 and Octadecylamine for a Sensitive Biosensor.

Controlling the assembly of DNA in order on a suitable electrode surface is of great significance for biosensors and disease diagnosis, but it is full of challenges. In this work, we creatively assembled DNA on the surface of octadecylamine (ODA)-modified topological insulator (Tls) Bi2Se3 and developed an electrochemical biosensor to detect biomarker DNA of coronavirus disease 2019 (COVID-19). A high-quality Bi2Se3 sheet was obtained from a single crystal synthesized in our lab. A uniform ODA layer was coated in argon by chemical vapor deposition (CVD). We observed and analyzed the assembly and mechanism of single-strand DNA (ssDNA) and double-strand DNA (dsDNA) on the Bi2Se3 surface through atomic force microscopy (AFM) and molecular dynamics (MD) simulations. The electrochemical signal revealed that the biosensor based on the DNA/ODA/Bi2Se3 electrode has a wide linear detection range from 1.0 × 10-12 to 1.0 × 10-8 M, with the limit of detection as low as 5 × 10-13 M. Bi2Se3 has robust surface states and improves the electrochemical signal-to-noise ratio, while the uniform ODA layer guides high-density ordered DNA, enhancing the sensitivity of the biosensor. Our work demonstrates that the ordered DNA/ODA/Bi2Se3 electrode surface has great application potential in the field of biosensing and disease diagnosis.

Sensitive biosensors based on topological insulator Bi2Se3 and peptide.

In this work, we designed a facile and label-free electrochemical biosensor based on intrinsic topological insulator (TI) Bi2Se3 and peptide for the detection of immune checkpoint molecules. With topological protection, Bi2Se3 could have robust surface states with low electronic noise, which was beneficial for the stable and sensitive electron transport between electrode and electrolyte interface. The peptides are easily synthesized and chemically modified, and have good biocompatibility and bioavailability, which is a suitable candidate as the recognition units for immune checkpoint molecules. Therefore, the peptide/Bi2Se3 was developed as a suitable working electrode for the electrochemical biosensor. The basic performance of the designed peptide/Bi2Se3 biosensor was investigated to determine the Anti-HA Tag Antibody and PD-L1 molecules. The linear detection range was from 3.6 × 10-10 mg mL-1 to 3.6 × 10-5 mg mL-1, and the detection limit was 1.07 × 10-11 mg mL-1. Moreover, the biosensor also displayed good selectivity and stability.

Topological insulator Bi2Se3 based electrochemical aptasensors for the application of sensitive detection of interferon-γ.

Interferon-γ (IFN-γ) is one of the crucial inflammatory cytokines as an early indicator of multiple diseases. A fast, simple, sensitive and reliable IFN-γ detection method is valuable for early diagnosis and monitoring of treatment. In this work, we creatively developed an electrochemical aptasensor based on the topological material Bi2Se3 for sensitive IFN-γ quantification. The high-quality Bi2Se3 sheet was directly exfoliated from a single crystal, which immobilized the synthesized IFN-γ aptamer. Under optimal conditions, the electrochemical signal revealed a wide linear relation along with the logarithmic concentration of IFN-γ from 1.0 pg mL-1 to 100.0 ng mL-1, with the limit of detection as low as 0.5 pg mL-1. The topological material Bi2Se3 with Dirac surface states improved the electrochemical signal/noise ratio and thus the sensitivity of the sensors. Furthermore, this electrochemical aptasensor exhibited excellent specificity and stability, which could be attributed to the large-scale smooth surface of the Bi2Se3 sheet with few defects decreasing the non-specific absorption. The developed biosensor has the same good performance as the ELISA method for detecting the real serum samples. Our work demonstrates that the developed electrochemical aptasensors based on topological materials have great potential in the field of clinical determination.

Lysosomal trafficking regulator restricts intracellular growth of Coxiella burnetii by inhibiting the expansion of Coxiella-containing vacuole and upregulating nos2 expression.

Coxiella burnetii is an obligate intracellular bacterium that causes Q fever, a zoonotic disease typically manifests as a severe flu-illness. After invading into the host cells, C. burnetii delivers effectors to regulate the vesicle trafficking and fusion events to form a large and mature Coxiella-containing vacuole (CCV), providing sufficient space and nutrition for its intracellular growth and proliferation. Lysosomal trafficking regulator (LYST) is a member of the Beige and Chediak-Higashi syndrome (BEACH) family, which regulates the transport of vesicles to lysosomes and regulates TLR signaling pathway, but the effect of LYST on C. burnetii infection is unclear. In this study, a series of experiments has been conducted to investigate the influence of LYST on intracellular growth of C. burnetii. Our results showed that lyst transcription was up-regulated in the host cells after C. burnetii infection, but there is no significant change in lyst expression level after infection with the Dot/Icm type IV secretion system (T4SS) mutant strain, while CCVs expansion and significantly increasing load of C. burnetii appeared in the host cells with a silenced lyst gene, suggesting LYST inhibits the intracellular proliferation of C. burnetii by reducing CCVs size. Then, the size of CCVs and the load of C. burnetii in the HeLa cells pretreated with E-64d were significantly decreased. In addition, the level of iNOS was decreased significantly in LYST knockout THP-1 cells, which was conducive to the intracellular replication of C. burnetii. This data is consistent with the phenotype of L-NMMA-treated THP-1 cells infected with C. burnetii. Our results revealed that the upregulation of lyst transcription after infection is due to effector secretion of C. burnetii and LYST inhibit the intracellular replication of C. burnetii by reducing the size of CCVs and inducing nos2 expression.

Simultaneous Cross-type Detection of Water Quality Indexes via a Smartphone-App Integrated Microfluidic Paper-Based Platform.

Water quality guarantee in remote areas necessitates the development of portable, sensitive, fast, cost-effective, and easy-to-use water quality detection methods. The current work reports on a microfluidic paper-based analytical device (µPAD) integrated with a smartphone app for the simultaneous detection of cross-type water quality parameters including pH, Cu(II), Ni(II), Fe(III), and nitrite. The shapes, baking time, amount, and ratios of reaction reagent mixtures of wax µPAD were optimized to improve the color uniformity and intensity effectively. An easy-to-use smartphone app was established for recording, analyzing, and directly reading the colorimetric signals and target concentrations on µPAD. The results showed that under the optimum conditions, the current analytical platform has reached the detection limits of 0.4, 1.9, 2.9, and 1.1 ppm for nitrite, Cu(II), Ni(II), and Fe(III), respectively, and the liner ranges are 2.3-90 ppm (nitrite), 3.8-400 ppm (Cu(II)), 2.9-1000 ppm (Ni(II)), 2.8-500 ppm (Fe(III)), and 5-9 (pH). The proposed portable smartphone-app integrated µPAD detection system was successfully applied to real industrial wastewater and river water quality monitoring. The proposed method has great potential for field water quality detection.

Water interaction with B-site (B = Al, Zr, Nb, and W) doped SrFeO3-δ-based perovskite surfaces for thermochemical water splitting applications.

Density functional theory (DFT) calculations were performed to study the interaction of water with the SrO and FeO2 terminations of the SrFeO3-δ (001) surface, where the effects of the metal dopants (Al, Zr, Nb, and W), surface oxygen vacancies, and oxygen ion migration were investigated. Our calculations showed that the metal dopants benefited the molecular and dissociative adsorptions of H2O on both the perfect and oxygen-vacancy-containing surfaces. The surface oxygen vacancies were predicted to promote the dissociative adsorption of H2O and the formation of H2. For all structures studied, H2 release was found to be always an overall endothermic process, except for the W-doped structure which will become exothermic at high temperature. On the oxygen-vacancy-containing surface, H2 generation was predicted to be easier at the SrO termination than the FeO2 termination. Furthermore, we also investigated the oxygen ion migration mechanism on all surface structures, predicted the behaviour of oxygen migration and the effect of oxygen vacancy defects. Our results showed that Al doping facilitated not only the formation of surface oxygen vacancies, but also oxygen migration from the surface to the subsurface, in contrast to the Zr, Nb and W-doped structures. This study provided significant insights into the interaction of water with the surfaces of doped SrFeO3-δ perovskite materials for thermochemical water splitting applications.

Mesoscopic sliding ferroelectricity enabled photovoltaic random access memory for material-level artificial vision system.

Intelligent materials with adaptive response to external stimulation lay foundation to integrate functional systems at the material level. Here, with experimental observation and numerical simulation, we report a delicate nano-electro-mechanical-opto-system naturally embedded in individual multiwall tungsten disulfide nanotubes, which generates a distinct form of in-plane van der Waals sliding ferroelectricity from the unique combination of superlubricity and piezoelectricity. The sliding ferroelectricity enables programmable photovoltaic effect using the multiwall tungsten disulfide nanotube as photovoltaic random-access memory. A complete "four-in-one" artificial vision system that synchronously achieves full functions of detecting, processing, memorizing, and powering is integrated into the nanotube devices. Both labeled supervised learning and unlabeled reinforcement learning algorithms are executable in the artificial vision system to achieve self-driven image recognition. This work provides a distinct strategy to create ferroelectricity in van der Waals materials, and demonstrates how intelligent materials can push electronic system integration at the material level.

A protein-protein interaction map reveals that the Coxiella burnetii effector CirB inhibits host proteasome activity.

Coxiella burnetii is the etiological agent of the zoonotic disease Q fever, which is featured by its ability to replicate in acid vacuoles resembling the lysosomal network. One key virulence determinant of C. burnetii is the Dot/Icm system that transfers more than 150 effector proteins into host cells. These effectors function to construct the lysosome-like compartment permissive for bacterial replication, but the functions of most of these effectors remain elusive. In this study, we used an affinity tag purification mass spectrometry (AP-MS) approach to generate a C. burnetii-human protein-protein interaction (PPI) map involving 53 C. burnetii effectors and 3480 host proteins. This PPI map revealed that the C. burnetii effector CBU0425 (designated CirB) interacts with most subunits of the 20S core proteasome. We found that ectopically expressed CirB inhibits hydrolytic activity of the proteasome. In addition, overexpression of CirB in C. burnetii caused dramatic inhibition of proteasome activity in host cells, while knocking down CirB expression alleviated such inhibitory effects. Moreover, we showed that a region of CirB that spans residues 91-120 binds to the proteasome subunit PSMB5 (beta 5). Finally, PSMB5 knockdown promotes C. burnetii virulence, highlighting the importance of proteasome activity modulation during the course of C. burnetii infection.

Electrochemical biosensor based on topological insulator Bi2Se3 tape electrode for HIV-1 DNA detection.

A large-size Bi2Se3 tape electrode (BTE) was prepared by peeling off a 2 × 1 × 0.5 cm high-quality single crystal. The feasibility of using the flexible BTE as an efficient bioplatform to load Au nanoparticles and probe DNA for HIV-1 DNA electrochemical sensing was explored. Differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS) show that the resultant biosensor has a wide linear range from 0.1 fM to 1 pM, a low detection limit of 50 aM, excellent selectivity, reproducibility and stability, and is superior to the pM DNA detection level of Pt-Au, graphene-AuNPs hybrid biosensors. This outstanding performance is attributed to the intrinsic surface state of Bi2Se3 topological insulator in facilitating electron transfer. Therefore, BTE electrochemical biosensor platform has great potential in the application for sensitive detection of DNA biomarkers.

Experimental and computational approaches to study the chlorination mechanism of pentlandite with ammonium chloride.

Pentlandite (Fe4.5Ni4.5S8) is the primary source for the metallurgical production of nickel worldwide, however it usually coexists with copper sulfide in nature. To develop an efficient and green process for the separation and extraction of valuable metals from the nickel sulfide concentrate, herein we conducted experimental studies and density functional theory (DFT) calculations to elucidate the chlorination mechanism of pentlandite using ammonium chloride (NH4Cl). First, low-temperature chlorination roasting experiments with NH4Cl were performed in which pentlandite was successfully converted into the corresponding metal chlorides (FeCl2 and NiCl2). Then, the chlorination product was analyzed via energy dispersive spectrometry to reveal the elemental distribution at the cross-section. Results reveal that Fe atoms in pentlandite underwent preferential chlorination to form a chloride layer, whereas Ni atoms remained at the center of the grain. Furthermore, density functional theory calculations were performed to investigate the chlorination mechanism of pentlandite by exploring two possible pathways, involving the adsorption of oxygen (O2), ammonium chloride (NH4Cl) and chlorine (Cl2) on both the (001) and (010) surfaces of pentlandite. Considering that the chlorination of pentlandite was achieved in air atmosphere, we first consider the direct chlorination of pentlandite by NH4Cl in the presence of oxygen. Dissociative oxygen adsorption was found to promote the chlorination process by providing oxygen sites for the dissociation of HCl, which is decomposed from NH4Cl, eventually leading to the formation of H2O and FeCl2 species. Alternatively, the reaction between pentlandite and Cl2 was proved to be feasible thermodynamically.

Development of a Lateral Flow Strip-Based Recombinase-Aided Amplification for Active Chlamydia psittaci Infection.

Chlamydia psittaci is the causative agent of psittacosis, a worldwide zoonotic disease. A rapid, specific, and sensitive diagnostic assay would be benefit for C. psittaci infection control. In this study, an assay combining recombinase-aided amplification and a lateral flow strip (RAA-LF) for the detection of active C. psittaci infection was developed. The RAA-LF assay targeted the CPSIT_RS02830 gene of C. psittaci and could be accomplished in 15 min at a single temperature (39°C). The analytical sensitivity of the assay was as low as 1 × 100 copies/µl and no cross-reaction with some other intracellular pathogens was observed. Moreover, all feces samples from mice infected with C. psittaci at day-1 post-infection were positive in the RAA-LF assay. In conclusion, the RAA-LF assay provides a convenient, rapid, specific and sensitive method for detection of active C. psittaci infection and it is also suitable for C. psittaci detection in field.