Sfxn5 Regulation of Actin Polymerization for Neutrophil Spreading Depends on a Citrate-Cholesterol-PI(4,5)P2 Pathway.

Cell spreading is an initial and critical step in neutrophil adhesion and migration, leading to neutrophil recruitment to inflammatory tissues. Sideroflexin (Sfxn) family proteins are metabolite transporters located in the mitochondrial membrane. Recombinant SFXN5 protein is a citrate transporter in vitro; however, whether Sfxn5 regulates any cellular behavior or function remains unknown. In this study, we found that small interfering RNA transfection or morpholino injection achieving Sfxn5 deficiency in neutrophils significantly decreased neutrophil recruitment in mice and zebrafish, respectively. Sfxn5 deficiency impaired neutrophil spreading and spreading-associated cellular phenotypes, such as cell adhesion, chemotaxis, and ROS production. Actin polymerization is critical for neutrophil spreading, and we found that actin polymerization in spreading neutrophils was partially inhibited by Sfxn5 deficiency. Mechanistically, we observed that the levels of cytosolic citrate and its downstream metabolic products, acetyl-CoA and cholesterol, were decreased in Sfxn5-deficient neutrophils. The levels of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), a mediator for the regulation of actin polymerization by cholesterol, were reduced in the plasma membrane of Sfxn5-deficient neutrophils. Exogenous supplementation with citrate or cholesterol partially reversed the reduction in PI(4,5)P2 levels, defective neutrophil actin polymerization, and cell spreading. Altogether, we demonstrated that Sfxn5 maintains cytosolic citrate levels and ensures the synthesis of sufficient cholesterol to promote actin polymerization in a PI(4,5)P2-dependent manner during neutrophil spreading, which is essential for the eventual inflammatory recruitment of neutrophils. Our study revealed the importance of Sfxn5 in neutrophil spreading and migration, thus identifying, to our knowledge, for the first time, the physiological cellular functions of the Sfxn5 gene.

Discovery of sulfone containing metallo-ß-lactamase inhibitors with reduced bacterial cell efflux and histamine release issues.

The design, syntheses and antibacterial evaluation of sulfone analogues of previously disclosed metallo-ß-lactamase inhibitors (MBLis) are described. The novel derivatives were overall more effective in gram-negative bacterial cell-based assays when combined with imipenem and relebactam. The major contributors to the improved anti-bacterial activity are enhanced enzyme-inhibitor interactions and reduced bacterial cell efflux monitored via an efflux assay involving isogenic Pseudomonas aeruginosa efflux + and efflux - tool strains.

A Novel Anion Receptor Additive for -40ºC Sodium Metal Batteries by Anion/Cation Solvation Engineering.

Sodium metal batteries, known for their high theoretical specific capacity, abundant reserves, and promising low-temperature performance, have garnered significant attention. However, the large ionic radius of Na+ and sluggish transport kinetics across the interfacial structure hinder their practical application. Previous reviews have rarely regulated electrolyte performance from the perspective of anions; as important components of the electrolyte, the regulation mechanism is not well understood. Herein, a novel anion receptor additive, 4-aminophenylboronic acid pinalol ester (ABAPE), is proposed to weaken the coupling between anions and cations and accelerate Na+ transport kinetics. The results of theoretical calculations and X-ray photoelectron spectroscopy with deep Ar-ion etching demonstrate that the introduction of this additive alters the solvation structure of Na+, reduces the desolvation barrier and forms a stable and dense electrode-electrolyte interface. Moreover, ABAPE forms hydrogen bonds (-NH···O/F) with H2O/HF, effectively preventing the hydrolysis of NaPF6 and stabilizing acidic species. Consequently, the Na||Na symmetric cell exhibits excellent long-cycle performance of 500 h at 1 mA cm-2 and 0.5 mAh cm-2. The Na||Na3V2(PO4)3 (NVP) full cell with the addition of ABAPE maintains a capacity retention of 84.29% at 1 C after 1200 cycles and presents no capacity decay over 150 cycles at -40°C.

Construction of Synchronous-Deposition K Metal Anodes Via Modulation of Ion/Electron Transport Kinetics for High-Rate and Low-Temperature K Metal Batteries.

The construction of conductive scaffolds is demonstrated to be an ideal strategy to alleviate the volume expansion and dendrite growth of K metal anodes. Nevertheless, the heterogeneous top-bottom deposition behavior caused by incompatible electronic/ionic conductivity of three-dimensional (3D) skeleton severely hinders its application. Here, a K2 Se/Cu conducting layer is fabricated on the Cu foam so as to enhance ionic transport and weaken electronic conductivity of the skeleton. Then, an excellent simultaneous deposition behavior of K metal inside the host is obtained for the first time via tuning fast ionic transport and low electronic conductivity. The simultaneous deposition mode can not only utilize the entire 3D structure to accommodate the volume expansion during K deposition but also avoid the formation of K dendrites at high current and ultra-low temperature. Consequently, the symmetric cells present a long cycle lifespan over 1000 h with a low deposition overpotential of 80 mV at 1 mA cm-2 . Furthermore, the full cell matching with the perylene-tetracarboxylic dianhydride (PTCDA) cathode presents an outstanding cycle lifespan over 600 cycles at 5 C at -20°C. The proposed simultaneous deposition strategy provides a new design direction for the construction of dendrite-free K metal anodes.

Optimized Solid Electrolyte Interphase and Solvation Structure of Potassium Ions in Carbonate Electrolytes for High-Performance Potassium Metal Batteries.

The introduction of electrolyte additives is one of the most potential strategies to improve the performance of potassium metal batteries (PMBs). However, designing an additive that can alter the K+ solvation shell and essentially inhibit K dendrite remains a challenge. Herein, the amyl-triphenyl-phosphonium bromide was introduced as an additive to build a stable solid electrolyte interphase layer. The amyl-TPP cations can form a cation shielding layer on the metal surface during the nucleation stage, preventing K+ from gathering at the tip to form K dendrites. Besides, the cations can be preferentially reduced to form Kx Py with fast K+ transport kinetics. The Br- anions, as Lewis bases with strong electronegativity, can not only coordinate the Lewis acid pentafluoride to inhibit the formation of HF, but also change the K+ solvation structure to reduce solvent molecules in the first solvation structure. Therefore, the symmetrical battery exhibits a low deposition overpotential of 123 mV at 0.1 mA cm-2 over 4200 h cycle life. The full battery, paried with a perylene-tetracarboxylic dianhydride (PTCDA) cathode, possesses a cycle life of 250 cycles at 2 C and 81.9% capacity retention. This work offers a reasonable electrolyte design to obtain PMBs with long-term stablity and safety.

Diversity of Fungal Endophytes in American Ginseng Seeds.

Seeds play a critical role in the production of American ginseng. Seeds are also one of the most important media for the long-distant dissemination and the crucial way for pathogen survival. Figuring out the pathogens carried by seeds is the basis for effective management of seedborne diseases. In this paper, we tested the fungi carried by the seeds of American ginseng from the main production areas of China using incubation and highly throughput sequencing methods. The seed-carried rates of fungi in Liuba, Fusong, Rongcheng, and Wendeng were 100, 93.8, 75.2, and 45.7%, respectively. Sixty-seven fungal species, which belonged to 28 genera, were isolated from the seeds. Eleven pathogens were identified from the seed samples. Among the pathogens, Fusarium spp. were found in all of the seed samples. The relative abundance of Fusarium spp. in the kernel was higher than that in the shell. Alpha index showed that the fungal diversity between seed shell and kernel differed significantly. Nonmetric multidimensional scaling analysis revealed that the samples from different provinces and between seed shell and kernel were distinctly separated. The inhibition rates of four fungicides to seed-carried fungi of American ginseng were 71.83% for Tebuconazole SC, 46.67% for Azoxystrobin SC, 46.08% for Fludioxonil WP, and 11.11% for Phenamacril SC. Fludioxonil, a conventional seed treatment agent, showed a low inhibitory effect on seed-carried fungi of American ginseng.

miR-143-3p represses leukemia cell proliferation by inhibiting KAT6A expression.

The present study is designed to investigate the expressions of microRNA-143-3p (miR-143-3p) and Lysine acetyltransferase 6A (KAT6A) in acute myeloid leukemia (AML) samples and AML cell lines and to explore the possible effects and underlying mechanisms of miR-143-3p on the proliferation of AML cells. The expressions of miR-143-3p and KAT6A in AML samples and cell lines were detected by RT-qPCR assay. CCK-8 and flow cytometry were performed to evaluate the role of KAT6A in viability of AML cells. EdU assay was performed to determine the effects of KAT6A on proliferation of AML cells. Western blot analysis was utilized to assess the impacts of KAT6A on proliferation-related protein expressions of AML cells. ELISA assay was adopted to illustrate the influence of KAT6A on inflammatory responses of AML cells. In addition, the relationship between KAT6A and miR-143-3p was predicted by ENCORI and miRWalk, and confirmed by dual-luciferase reporter assay. Moreover, the effects of KAT6A on the proliferation of AML cells mediated with miR-143-3p were carried out by rescue experiment. The expression of KAT6A was significantly upregulated, while miR-134-4p was downregulated both in the AML tissues and in AML cell lines. In addition, the silence of KAT6A significantly inhibited the viability of AML cells. Besides, KAT6A silencing notably suppressed the proliferation of AML cells and reduced the protein expressions of Ki-67 and PCNA. Knockdown of KAT6A notably decreased the expression levels of IL-1ß, TNF-α and IL-6, and increased the expression levels of TGF-ß and IL-10. Moreover, overexpression of miR-143-3p repressed viability and proliferation of AML cells and overexpression of KAT6A partially reversed the inhibitory effects of miR-143-3p mimic on viability and proliferation of AML cells. miR-143-3p/KAT6A played an essential role in the viability and proliferation of AML cells.

A Miniature Particle Mass Spectrometer.

Microparticles play important roles in our life. Besides chemical compositions and morphology, the size of microparticles will also decide their behavior in the environment or in organisms. Weighing the mass of microparticles by mass spectrometry is a useful method to characterize their size. In this technical note, a miniature particle mass spectrometer with an aerodynamic desorption/ionization ion source has been developed. We used a compact main control board to produce an ac voltage for trapping and ejecting the particles. The sampling process and data acquisition were also controlled by this board. We utilized this instrument to measure polystyrene spheres, silica particles, and mice red blood cells. Mass distributions of these particles were obtained rapidly with good accuracy.

Growth and Transfer of Seamless 3D Graphene-Nanotube Hybrids.

Seamlessly connected graphene and carbon nanotube hybrids (GCNTs) have great potential as carbon platform structures in electronics due to their high conductivity and high surface area. Here, we introduce a facile method for making patterned GCNTs and their intact transfer onto other substrates. The mechanism for selective growth of vertically aligned CNTs (VA-CNTs) on the patterned graphene is discussed. The complete transfer of the GCNT pattern onto other substrates is possible because of the mechanical strength of the GCNT hybrids. Electrical conductivity measurements of the transferred GCNT structures show Ohmic contact through the VA-CNTs to graphene--evidence of its integrity after the transfer process.

Discovery of a potent and selective ROMK inhibitor with improved pharmacokinetic properties based on an octahydropyrazino[2,1-c][1,4]oxazine scaffold.

Following the discovery of small molecule acyl piperazine ROMK inhibitors, the acyl octahydropyrazino[2,1-c][1,4]oxazine series was identified. This series displays improved ROMK/hERG selectivity, and as a consequence, the resulting ROMK inhibitors do not evoke QTc prolongation in an in vivo cardiovascular dog model. Further efforts in this series led to the discovery of analogs with improved pharmacokinetic profiles. This new series also retained comparable ROMK potency compared to earlier leads.

GENERATION OF TOLEROGENIC DENDRITIC CELLS UNDER THE PERSISTENT INFLAMMATION STIMULATION.

ABSTRACT: Immunosuppression, commonly accompanied by persistent inflammation, is a key feature in the later phase of sepsis. However, the pathophysiological mechanisms underlying this phenomenon remain unclear. Dendritic cells (DCs), specifically tolerogenic DCs (tolDCs), play a crucial role in this process by regulating immune responses through inducing T cell anergy and releasing anti-inflammatory cytokines. Nevertheless, the existing cell models are inadequate for investigating tolDCs during the immunosuppressive phase of sepsis. Therefore, this study aimed to develop a novel in vitro model to generate tolDCs under chronic inflammatory conditions. We have successfully generated tolDCs by exposing them to sublethal lipopolysaccharide (LPS) for 72 h while preserving cell viability. Considering that IL-10-induced tolDCs (IL-10-tolDCs) are well-established models, we compared the immunological tolerance between LPS-tolDCs and IL-10-tolDCs. Our findings indicated that both LPS-tolDCs and IL-10-tolDCs exhibited reduced expression of maturation markers, whereas their levels of inhibitory markers were elevated. Furthermore, the immunoregulatory activities of LPS-tolDCs and IL-10-tolDCs were found to be comparable. These dysfunctions include impaired antigen presenting capacity and suppression of T cell activation, proliferation, and differentiation. Notably, compared with IL-10-tolDCs, LPS-tolDCs showed a reduced response in maturation and cytokine production upon stimulation, indicating their potential as a better model for research. Overall, in comparison with IL-10-tolDCs, our data suggest that the immunological dysfunctions shown in LPS-tolDCs could more effectively elucidate the increased susceptibility to secondary infections during sepsis. Consequently, LPS-tolDCs have emerged as promising therapeutic targets for ameliorating the immunosuppressed state in septic patients.

Structure Guided Discovery of Novel Pan Metallo-ß-Lactamase Inhibitors with Improved Gram-Negative Bacterial Cell Penetration.

The use of ß-lactam (BL) and ß-lactamase inhibitor combination to overcome BL antibiotic resistance has been validated through clinically approved drug products. However, unmet medical needs still exist for the treatment of infections caused by Gram-negative (GN) bacteria expressing metallo-ß-lactamases. Previously, we reported our effort to discover pan inhibitors of three main families in this class: IMP, VIM, and NDM. Herein, we describe our work to improve the GN coverage spectrum in combination with imipenem and relebactam. This was achieved through structure- and property-based optimization to tackle the GN cell penetration and efflux challenges. A significant discovery was made that inhibition of both VIM alleles, VIM-1 and VIM-2, is essential for broad GN coverage, especially against VIM-producing P. aeruginosa. In addition, pharmacokinetics and nonclinical safety profiles were investigated for select compounds. Key findings from this drug discovery campaign laid the foundation for further lead optimization toward identification of preclinical candidates.

[Spectroscopic study of dilute magnetic semiconductors Zn(1-x) Fe(x)O nanoparticles].

Diluted magnetic semiconductors Zn(1-x) Fe(x)O nanoparticles with different content (x = 0, 0.01, 0.05, 0.10 and 0.20) were successfully synthesized via hydrothermal method. The X-ray diffraction (XRD) shows that the samples are wurtzite structure and metallic Fe or other secondary phases were not found in the samples. The transmission electron microscopy (TEM) shows that the morphology is nanoparticles with good dispersion, and the lattice is clearly visible. Raman scattering spectrum (Raman spectra) shows that E2 (High) mode broadened, shifted towards the high-frequencies side and decreased the peak intensity. Photoluminescence spectra (PL) shows that the peaks moved to lower energy and the photoluminescence intensity was quenched with increasing Fe doping concentration. The ultraviolet-visible spectrophotometry (UV-Vis) indicates that the optical band gap decreased and red shift occured. All the results indicate that Fe3+ ions successfully substituted for Zn2+ and were incorporated into the crystal lattice of ZnO.

Rapid Identification of Chinese Hamster Ovary Cell Apoptosis and Its Potential Role in Process Robustness Assessment.

Currently, the assessment of process robustness is often time-consuming, labor-intensive, and material-intensive using process characterization studies. Therefore, a simple and time-saving method is highly needed for the biopharmaceutical industry. Apoptosis is responsible for 80% of Chinese hamster ovary (CHO) cell deaths and affects the robustness of the cell culture process. This study's results showed that a more robust process can support cells to tolerate apoptosis for a longer time, suggesting that the robustness of the process could be judged by the ability of cells to resist apoptosis. Therefore, it is necessary to establish a rapid method to detect the apoptosis of CHO cells. In trying to establish a new method for detecting apoptosis in large-scale cell cultures, glucose withdrawal was studied, and the results showed that CHO cells began to apoptose after glucose was consumed. Then, the concentration of extracellular potassium increased, and a prolongation of apoptosis time was observed. Further study results showed that the process with poor robustness was associated with a higher proportion of apoptosis and extracellular potassium concentration, so potassium could be used as a biochemical index of apoptosis. The strategy we present may be used to expedite the assessment of process robustness to obtain a robust cell culture process for other biologics.

Shear Performance of Epoxy Joints in a Precast Bridge Deck Considering Constraint Effects.

The joint form plays a vital role in the rapid assembly of precast bridge decks for steel-concrete composite bridges. Existing research primarily focuses on studying the shear performance of joints through direct shear tests, which is insufficient to fully reflect the mechanical behavior of joints under the constraint of prefabricated bridge deck panels during actual vehicular traffic. Considering situations such as vehicle loads and external forces acting on precast bridge decks, this study investigates the shear performance of epoxy joints under constraint through an improved shear test. The influence of constraint force, shear key details and interface defects on the shear performance of epoxy joints is investigated. The results reveal that the shear test method employed in this study can realistically reflect the shear performance of epoxy joints in precast bridge decks. Both active and passive constrained epoxy joint specimens exhibited no interface cracks, and their failure modes were identified as shear failure between mid-span supports. Compared with passive constraint, the shear-bearing capacity of epoxy joint specimens under active constraint was increased by 86.1~130.6%. Among the epoxy joint specimens with depth-height ratios of 15/110, 25/110, 35/110 and 45/110, the joint with a depth of 35 mm demonstrated the highest shear strength. Furthermore, the shear performance of epoxy joints significantly deteriorated when the interface defects exceeded 30%, resulting in the failure mode transforming from shear failure to interface failure.

KMT2A is targeted by miR-361-3p and modulates leukemia cell's abilities to proliferate, migrate and invade.

OBJECTIVE: The lives and safety of humans are significantly threatened by acute myeloid leukemia (AML), which is proven to be the most prevalent acute leukemia. This work is therefore intended to investigate and analyze the expressions of miR-361-3p and Histone Lysine Methyltransferase 2A (KMT2A) in tissues and cell lines of AML and identify an advanced and novel target for the therapy of AML. METHODS: The qRT-PCR and western blot assays were conducted to find expressions of miR-361-3p/KMT2A in AML PB and cell lines. After then, tests using CCK-8 and EdU were run to see how KMT2A affected the growth of AML cells. Transwell migration and invasion assay was conducted to evaluate KMT2A's contribution to the migration and invasion of AML cells. ENCORI and miRWalk predicted the association between KMT2A and miR-361-3p, and the dual-luciferase reporter experiment verified it. Furthermore, rescue studies were used to ascertain how KMT2A affected the miR-361-3p-regulated AML cells' abilities to proliferate, migrate, and invade. RESULTS: miR-361-3p was poorly expressed while KMT2A was abundantly expressed. Additionally, KMT2A downregulation prevented AML cells from proliferating. PCNA and Ki-67 protein levels fell when KMT2A was silent. Furthermore, AML cells' motility, invasion, and metastasis were inhibited by low KMT2A expression. KMT2A was also identified as a direct target of miR-361-3p and negatively correlated with miR-361-3p. Finally, the over-expression of KMT2A partially reversed the inhibitory effects of up-regulation of miR-361-3p. CONCLUSION: A potential therapeutic candidate target for the treatment of AML may be miR-361-3p/KMT2A.

Progress of Metal Nanomaterial Controllable Preparation by Photoreduction.

Metal nanoparticles (NPs) are widely used in biomedicine, catalysis, environment, electronics, and other fields, which is closely related to its structural form. For this purpose, researchers have been looking for a simple, green, and controllable way to mass produce metal nanomaterials with desired characteristics (shape, size, stability, etc.). Due to the surface plasmon resonance (SPR) effect of metal nanoparticles, photoreduction method can control the morphology of metal nanoparticles well, which is also simple, large-scalable, and energy-saving. This review provides an overview of the photoreduction method for the synthesis of metal nanoparticles and discusses the factors such as the light source, pH value, reagents, and temperature on the morphology of the nanoparticles. Finally, the challenges and development trends in the controlled preparation of nanomaterials are proposed.

Dynamic increase in myoglobin level is associated with poor prognosis in critically ill patients: a retrospective cohort study.

Background: Myoglobin is an important biomarker for monitoring critically ill patients. However, the relationship between its dynamic changes and prognosis remains unclear. Methods: We retrospectively enrolled 11,218 critically ill patients from a general and surgical intensive care unit (ICU) of a tertiary hospital between June 2016 and May 2020. Patients with acute cardiovascular events, cardiac and major vascular surgeries, and rhabdomyolysis were excluded. To investigate the early myoglobin distribution, the critically ill patients were stratified according to the highest myoglobin level within 48 h after ICU admission. Based on this, the critically ill patients with more than three measurements within 1 week after ICU admission were included, and latent class trajectory modeling was used to classify the patients. The characteristics and outcomes were compared among groups. Sensitivity analysis was performed to exclude patients who had died within 72 h after ICU admission. Restricted mean survival time regression model based on pseudo values was used to determine the 28-day relative changes in survival time among latent classes. The primary outcome was evaluated with comparison of in-hospital mortality among each Trajectory group, and the secondary outcome was 28-day mortality. Results: Of 6,872 critically ill patients, 3,886 (56.5%) had an elevated myoglobin level (≥150 ng/mL) at admission to ICU, and the in-hospital mortality significantly increased when myoglobin level exceeded 1,000 µg/mL. In LCTM, 2,448 patients were unsupervisedly divided into four groups, including the steady group (n = 1,606, 65.6%), the gradually decreasing group (n = 523, 21.4%), the slowly rising group (n = 272, 11.1%), and the rapidly rising group (n = 47, 1.9%). The rapidly rising group had the largest proportion of sepsis (59.6%), the highest median Sequential Organ Failure Assessment (SOFA) score (10), and the highest in-hospital mortality (74.5%). Sensitivity analysis confirmed that 98.2% of the patients were classified into the same group as in the original model. Compared with the steady group, the rapidly rising group and the slowly rising group were significantly related to the reduction in 28-day survival time (ß = -12.08; 95% CI -15.30 to -8.86; ß = -4.25, 95% CI -5.54 to -2.97, respectively). Conclusion: Elevated myoglobin level is common in critically ill patients admitted to the ICU. Dynamic monitoring of myoglobin levels offers benefit for the prognosis assessment of critically ill patients.

Identification of histone deacetylase inhibitors as neutrophil recruitment modulators in zebrafish using a chemical library screen.

Tissue injury-induced neutrophil recruitment is a prerequisite for the initiation and amplification of inflammatory responses. Although multiple proteases and enzymes involved in post-translational modification (PTM) of proteins regulate leukocyte recruitment, an unbiased functional screen of enzymes regulating inflammatory leukocyte recruitment has yet to be undertaken. Here, using a zebrafish tail fin amputation (TFA) model to screen a chemical library consisting of 295 compounds that target proteases and PTM enzymes, we identified multiple histone deacetylase (HDAC) inhibitors that modulate inflammatory neutrophil recruitment. AR-42, a pan-HDAC inhibitor, was shown to inhibit neutrophil recruitment in three different zebrafish sterile tissue injury models: a TFA model, a copper-induced neuromast damage and mechanical otic vesicle injury (MOVI) model, and a sterile murine peritonitis model. RNA sequencing analysis of AR-42-treated fish embryos revealed downregulation of neutrophil-associated cytokines/chemokines, and exogenous supplementation with recombinant human IL-1ß and CXCL8 partially restored the defective neutrophil recruitment in AR-42-treated MOVI model fish embryos. We thus demonstrate that AR-42 non-cell-autonomously modulates neutrophil recruitment by suppressing transcriptional expression of cytokines/chemokines, thereby identifying AR-42 as a promising anti-inflammatory drug for treating sterile tissue injury-associated diseases.