Pseudomonas aeruginosa two-component system CprRS regulates HigBA expression and bacterial cytotoxicity in response to LL-37 stress.

Pseudomonas aeruginosa is a highly pathogenic bacterium known for its ability to sense and coordinate the production of virulence factors in response to host immune responses. However, the regulatory mechanisms underlying this process have remained largely elusive. In this study, we investigate the two-component system CprRS in P. aeruginosa and unveil the crucial role of the sensor protein CprS in sensing the human host defense peptide LL-37, thereby modulating bacterial virulence. We demonstrate that CprS acts as a phosphatase in the presence of LL-37, leading to the phosphorylation and activation of the response regulator CprR. The results prove that CprR directly recognizes a specific sequence within the promoter region of the HigBA toxin-antitoxin system, resulting in enhanced expression of the toxin HigB. Importantly, LL-37-induced HigB expression promotes the production of type III secretion system effectors, leading to reduced expression of proinflammatory cytokines and increased cytotoxicity towards macrophages. Moreover, mutations in cprS or cprR significantly impair bacterial survival in both macrophage and insect infection models. This study uncovers the regulatory mechanism of the CprRS system, enabling P. aeruginosa to detect and respond to human innate immune responses while maintaining a balanced virulence gene expression profile. Additionally, this study provides new evidence and insights into the complex regulatory system of T3SS in P. aeruginosa within the host environment, contributing to a better understanding of host-microbe communication and the development of novel strategies to combat bacterial infections.

Molecular mechanism of the one-component regulator RccR on bacterial metabolism and virulence.

The regulation of carbon metabolism and virulence is critical for the rapid adaptation of pathogenic bacteria to host conditions. In Pseudomonas aeruginosa, RccR is a transcriptional regulator of genes involved in primary carbon metabolism and is associated with bacterial resistance and virulence, although the exact mechanism is unclear. Our study demonstrates that PaRccR is a direct repressor of the transcriptional regulator genes mvaU and algU. Biochemical and structural analyses reveal that PaRccR can switch its DNA recognition mode through conformational changes triggered by KDPG binding or release. Mutagenesis and functional analysis underscore the significance of allosteric communication between the SIS domain and the DBD domain. Our findings suggest that, despite its overall structural similarity to other bacterial RpiR-type regulators, RccR displays a more complex regulatory element binding mode induced by ligands and a unique regulatory mechanism.

Pyrogenic Carbon Degradation by Galvanic Coupling with Sprayed Seawater Microdroplets.

Surface waves are known for their mechanical role in coastal processes that influence the weather and climate. However, their chemical impact, particularly on the transformation of pyrogenic carbon, is poorly understood. Pyrogenic carbon is generally assumed to show negligible postformational alteration of its stable carbon isotope composition. Here we present an electrochemical interaction of pyrogenic carbon with the sprayed seawater microdroplets resulting from wave breaking, driven by the galvanic coupling between the microdroplet water-carbon interfaces and the microdroplet water-vapor interfaces. This enables refractory pyrogenic carbon to rapidly degrade via the oxygenation and mineralization reaction, which makes it ∼2.6‰ enriched in 13C, far exceeding the generally assumed postformation alteration values (<0.5‰) of pyrogenic carbon. The unique chemical dynamics of seawater microdroplets provide new insights into the discrepancy in carbon isotope signatures between riverine and marine black carbon, emphasizing the potential of coastal oceans for carbon sequestration in the global carbon cycle.

Shoc2 recognizes bacterial flagellin and mediates antibacterial Erk/Stat signaling in an invertebrate.

Flagellin is a key bacterial virulence factor that can stimulate molecular immune signaling in both animals and plants. The detailed mechanisms of recognizing flagellin and mounting an efficient immune response have been uncovered in vertebrates; however, whether invertebrates can discriminate flagellin remains largely unknown. In the present study, the homolog of human SHOC2 leucine rich repeat scaffold protein in kuruma shrimp (Marsupenaeus japonicus), designated MjShoc2, was found to interact with Vibrio anguillarum flagellin A (FlaA) using yeast two-hybrid and pull-down assays. MjShoc2 plays a role in antibacterial response by mediating the FlaA-induced expression of certain antibacterial effectors, including lectin and antimicrobial peptide. FlaA challenge, via MjShoc2, led to phosphorylation of extracellular regulated kinase (Erk), and the subsequent activation of signal transducer and activator of transcription (Stat), ultimately inducing the expression of effectors. Therefore, by establishing the FlaA/MjShoc2/Erk/Stat signaling axis, this study revealed a new antibacterial strategy in shrimp, and provides insights into the flagellin sensing mechanism in invertebrates.

Conformational remodeling enhances activity of lanthipeptide zinc-metallopeptidases.

Lanthipeptides are an important group of natural products with diverse biological functions, and their biosynthesis requires the removal of N-terminal leader peptides (LPs) by designated proteases. LanPM1 enzymes, a subgroup of M1 zinc-metallopeptidases, have been recently identified as bifunctional proteases with both endo- and aminopeptidase activities to remove LPs of class III and class IV lanthipeptides. Herein, we report the biochemical and structural characterization of EryP as the LanPM1 enzyme from the biosynthesis of class III lanthipeptide erythreapeptin. We determined X-ray crystal structures of EryP in three conformational states, the open, intermediate and closed states, and identified a unique interdomain Ca2+ binding site as a regulatory element that modulates its domain dynamics and proteolytic activity. Inspired by this regulatory Ca2+ binding, we developed a strategy to engineer LanPM1 enzymes for enhanced catalytic activities by strengthening interdomain associations and driving the conformational equilibrium toward their closed forms.

The novel type II toxin-antitoxin PacTA modulates Pseudomonas aeruginosa iron homeostasis by obstructing the DNA-binding activity of Fur.

Type II toxin-antitoxin (TA) systems are widely distributed in bacterial and archaeal genomes and are involved in diverse critical cellular functions such as defense against phages, biofilm formation, persistence, and virulence. GCN5-related N-acetyltransferase (GNAT) toxin, with an acetyltransferase activity-dependent mechanism of translation inhibition, represents a relatively new and expanding family of type II TA toxins. We here describe a group of GNAT-Xre TA modules widely distributed among Pseudomonas species. We investigated PacTA (one of its members encoded by PA3270/PA3269) from Pseudomonas aeruginosa and demonstrated that the PacT toxin positively regulates iron acquisition in P. aeruginosa. Notably, other than arresting translation through acetylating aminoacyl-tRNAs, PacT can directly bind to Fur, a key ferric uptake regulator, to attenuate its DNA-binding affinity and thus permit the expression of downstream iron-acquisition-related genes. We further showed that the expression of the pacTA locus is upregulated in response to iron starvation and the absence of PacT causes biofilm formation defect, thereby attenuating pathogenesis. Overall, these findings reveal a novel regulatory mechanism of GNAT toxin that controls iron-uptake-related genes and contributes to bacterial virulence.

Fluorescence sensor based on optimized quantum yield manganese-carbon polymer dots and smartphone-integrated sensing platform for tetracycline detection.

The one-step synthesis of Mn-doped carbon quantum dots (Mn-CPDs) with a high quantum yield (QY = 45%) is reported using the microwave-assisted method. Subsequently, Mn-CPDs were successfully combined with Eu3+ ions to construct an Eu3+@Mn-CPDs fluorescence sensor. The presence of tetracycline (TC) induced a transition of fluorescence emission from blue (434 nm) to red (618 nm), and a robust linear relationship was observed between the ratio of F618 nm / F434 nm and the TC concentration (5 - 50 nmol/L), with a limit of detection (LOD) of 5.76 nmol/L. The underlying mechanism of Eu3+@Mn-CPDs and TC sensing is unveiled as a synergistic effect involving inner filter effect (IFE) and concurrent interactions. Notably, the smartphone-integrated sensing platform based on Eu3+@Mn-CPDs enables rapid and quantitative TC detection within a short time (< 30 s) by monitoring fluorescence color changes, achieving high-detection sensitivities (with a LOD of 6.18 nmol/L). This versatile and efficient sensing platform demonstrates its potential for the determination of TC concentrations in milk, honey, and tap water samples.

An automated method for assessing condyle head changes in patients with skeletal class II malocclusion based on Cone-beam CT images.

OBJECTIVES: Currently, there is no reliable automated measurement method to study the changes in the condylar process after orthognathic surgery. Therefore, this study proposes an automated method to measure condylar changes in patients with skeletal class II malocclusion following surgical-orthodontic treatment. METHODS: Cone-beam CT (CBCT) scans from 48 patients were segmented using the nnU-Net network for automated maxillary and mandibular delineation. Regions unaffected by orthognathic surgery were selectively cropped. Automated registration yielded condylar displacement and volume calculations, each repeated three times for precision. Logistic regression and linear regression were used to analyse the correlation between condylar position changes at different time points. RESULTS: The Dice score for the automated segmentation of the condyle was 0.971. The intraclass correlation coefficients (ICCs) for all repeated measurements ranged from 0.93 to 1.00. The results of the automated measurement showed that 83.33% of patients exhibited condylar resorption occurring six months or more after surgery. Logistic regression and linear regression indicated a positive correlation between counterclockwise rotation in the pitch plane and condylar resorption (P < .01). And a positive correlation between the rotational angles in both three planes and changes in the condylar volume at six months after surgery (P ≤ .04). CONCLUSIONS: This study's automated method for measuring condylar changes shows excellent repeatability. Skeletal class II malocclusion patients may experience condylar resorption after bimaxillary orthognathic surgery, and this is correlated with counterclockwise rotation in the sagittal plane. ADVANCES IN KNOWLEDGE: This study proposes an innovative multi-step registration method based on CBCT, and establishes an automated approach for quantitatively measuring condyle changes post-orthognathic surgery. This method opens up new possibilities for studying condylar morphology.

Three-dimensional echo light field analysis for dual-band laser active detection of a cat-eye optical system.

Laser active detection technology utilizing the cat-eye effect provides rapid response, precise positioning, and long detection distances. However, current research mainly focuses on active detection within a single visible or near-infrared band, lacking quantitative analyses of the echo spot. In this paper, a four-interval theoretical model for dual band cat-eye target echo detection was constructed using matrix optics theory and Collins diffraction integration method. Dual-band echo detection experiments were conducted using 10.6 um far-infrared waves and 532 nm visible light waves, also the power, radius, and target-missing quantities of the echo spots were collected and quantitatively compared with the theoretical results. Results indicate that, due to the diffraction limit's effect on the distribution of the echo field, the echo power of far-infrared band detection is smaller than that of visible light band detection. The impact on the light spot caused by the positive and negative defocus values is asymmetric, with positive defocus having a lower impact on the echo spot than negative defocus at the same value. A weak positive defocus value that minimizes the radius of the echo spot and maximizes the echo power exists, with the value of weak positive defocus varying between detection bands. A linear relationship exists between the incident angle of the detection laser and the deviation of the echo spot. These findings provide a foundation for extracting working band details, predicting the motion trajectory of moving cat-eye targets, and achieving real-time tracking and detection recognition during laser active detection.

On-chip coherent beam combination of waveguide amplifiers on Er3+-doped thin film lithium niobate.

We demonstrate on-chip coherent beam combination of two waveguide amplifiers on Er3+-doped thin film lithium niobate (Er:TFLN) platform. Our device is built based on an electro-optic modulator fabricated on Er:TFLN. The output power of the coherently combined amplifiers is measured as high as 12.9 mW, surpassing that of previous single waveguide amplifiers based on an Er3+-doped thin film lithium niobate platform.

The prognostic significance of clinicopathological characteristics in early-onset versus late-onset colorectal cancer liver metastases.

PURPOSE: This study aimed to explore the prognostic significance of clinicopathological characteristics in early-onset versus late-onset colorectal liver metastases (CRLM). METHODS: The data of CRLM patients who underwent hepatectomy from September 2010 to September 2020 were retrospectively analyzed. According to the age of primary cancer diagnosis, patients were divided into early-onset CRLM (EOCRLM) and late-onset CRLM (LOCRLM) groups. Clinicopathological parameters were compared between the two groups. Cox regression model and Kaplan-Meier method were used to analyze the effect of clinicopathological parameters on overall survival (OS) and recurrence-free survival (RFS). RESULTS: In total, 431 CRLM patients were identified, 130 with EOCRLM and 301 with LOCRLM. Compared with LOCRLM patients, EOCRLM patients had lower American Society of Anesthesia (ASA) grade and longer operation time (204 vs. 179 min). More aggressive features were presented in EOCRLM patients including synchronous liver metastases (76.9% vs. 61.1%) and bilobar involvement (43.8% vs. 33.2%). No significant difference in OS or RFS was found between the two groups. Multivariate analysis of EOCRLM group showed that preoperative CA19-9 level and RAS/BRAF status were predictive of OS, while bilobar involvement and preoperative CEA level were associated with RFS. In LOCRLM group, the number of CRLM, preoperative CA19-9 level, and BRAF status were associated with OS, while the number of CRLM was associated with RFS. CONCLUSIONS: The preoperative CA19-9 level, RAS/BRAF status, bilobar involvement, and preoperative CEA level were predictive of EOCRLM patient prognosis, while the number of CRLM, preoperative CA19-9 level, and BRAF status were predictive of LOCRLM patient prognosis.

Fatigue behavior of zirconia with microgrooved surfaces produced using femtosecond laser.

Femtosecond laser is a promising surface treatment tool for zirconia implant. In this study, the fatigue behavior of zirconia specimens with microgrooved surfaces formed by femtosecond laser is reported. One hundred sixty CAD/CAM zirconia bars (20 mm × 4 mm × 1.4 mm) were evenly divided into four groups with different surface: as sintered; sandblasted with 110 µm Al2O3; femtosecond laser produced microgrooves having 50 µm width, 30 µm depth, and 100 µm pitch; microgrooves having 30 µm width, 20 µm depth, and 60 µm pitch. The femtosecond laser formed micro/nanostructured microgrooves with precise size on zirconia surfaces. XRD analysis indicated that microgrooved surface showed no obvious tetragonal-to-monoclinic phase transformation. The fatigue strength of sandblasted specimens (728 MPa) was significantly higher than that of as sintered specimens (570 MPa). However, the fatigue strength of specimens with microgrooved surface decreased to about 360-380 MPa. The results suggest femtosecond laser is an effective technique to regulate the surface microtopography of zirconia, while further investigations are needed to improve its fatigue behavior.

Designing Thiophene-Enriched Fully Conjugated 3D Covalent Organic Framework as Metal-Free Oxygen Reduction Catalyst for Hydrogen Fuel Cells.

The application of three-dimensional (3D) covalent organic frameworks (COFs) in renewable energy fields is greatly limited due to their non-conjugated skeletons. Here, we design and successfully synthesize a thiophene-enriched fully conjugated 3D COF (BUCT-COF-11) through an all-thiophene-linked saddle-shaped building block (COThTh-CHO). The BUCT-COF-11 exhibits excellent semiconducting property with intrinsic metal-free oxygen reduction reaction (ORR) activity. Using the COF as cathode catalyst, the assembled anion-exchange membrane fuel cells (AEMFCs) exhibited a high peak power density up to 493 mW cm-2 . DFT calculations reveal that thiophene introduction in the COF not only improves the conductivity but also optimizes the electronic structure of the sample, which therefore boosts the ORR performance. This is the first report on the application of COFs as metal-free catalysts in fuel cells, demonstrating the great potential of fully conjugated 3D COFs as promising semiconductors in energy fields.

Synergistic Effect of Halogen Ions and Shelling Temperature on Anion Exchange Induced Interfacial Restructuring for Highly Efficient Blue Emissive InP/ZnS Quantum Dots.

InP quantum dots (QDs) have attracted much attention owing to their nontoxic properties and shown great potential in optoelectronic applications. Due to the surface defects and lattice mismatch, the interfacial structure of InP/ZnS QDs plays a significant role in their performance. Herein, the formation of In-S and Sx -In-P1-x interlayers through anion exchange at the shell-growth stage is revealed. More importantly, it is proposed that the composition of interface is dependent on the synergistic effect of halogen ions and shelling temperature. High shelling temperature contributes to the optical performance improvement resulting from the formation of interlayers, besides the thicker ZnS shell. Moreover, the effect relates to the halogen ions where I- presents more obvious enhancement than Br- and Cl- , owing to their different ability to coordinate with In dangling bonds, which are inclined to form In-S and Sx -In-P1-x bonds. Further, the anion exchange under I- -rich environment causes a blue-shift of emission wavelength with shelling temperature increasing, unobserved in a Cl- - or Br- -rich environment. It contributes to the preparation of highly efficient blue emissive InP/ZnS QDs with emission wavelength of 473 nm, photoluminescence quantum yield of ≈50% and full width at half maximum of 47 nm.

Recent Advances in Multiresponsive Flexible Sensors towards E-skin: A Delicate Design for Versatile Sensing.

Multiresponsive flexile sensors with strain, temperature, humidity, and other sensing abilities serving as real electronic skin (e-skin) have manifested great application potential in flexible electronics, artificial intelligence (AI), and Internet of Things (IoT). Although numerous flexible sensors with sole sensing function have already been reported since the concept of e-skin, that mimics the sensing features of human skin, was proposed about a decade ago, the ones with more sensing capacities as new emergences are urgently demanded. However, highly integrated and highly sensitive flexible sensors with multiresponsive functions are becoming a big thrust for the detection of human body motions, physiological signals (e.g., skin temperature, blood pressure, electrocardiograms (ECG), electromyograms (EMG), sweat, etc.) and environmental stimuli (e.g., light, magnetic field, volatile organic compounds (VOCs)), which are vital to real-time and all-round human health monitoring and management. Herein, this review summarizes the design, manufacturing, and application of multiresponsive flexible sensors and presents the future challenges of fabricating these sensors for the next-generation e-skin and wearable electronics.

Chaperone-tip adhesin complex is vital for synergistic activation of CFA/I fimbriae biogenesis.

Colonization factor CFA/I defines the major adhesive fimbriae of enterotoxigenic Escherichia coli and mediates bacterial attachment to host intestinal epithelial cells. The CFA/I fimbria consists of a tip-localized minor adhesive subunit, CfaE, and thousands of copies of the major subunit CfaB polymerized into an ordered helical rod. Biosynthesis of CFA/I fimbriae requires the assistance of the periplasmic chaperone CfaA and outer membrane usher CfaC. Although the CfaE subunit is proposed to initiate the assembly of CFA/I fimbriae, how it performs this function remains elusive. Here, we report the establishment of an in vitro assay for CFA/I fimbria assembly and show that stabilized CfaA-CfaB and CfaA-CfaE binary complexes together with CfaC are sufficient to drive fimbria formation. The presence of both CfaA-CfaE and CfaC accelerates fimbria formation, while the absence of either component leads to linearized CfaB polymers in vitro. We further report the crystal structure of the stabilized CfaA-CfaE complex, revealing features unique for biogenesis of Class 5 fimbriae.

Maturation of an Antimicrobial Peptide Inhibits Aeromonas hydrophila Infection in Crayfish.

Rapid synthesis and release of active antimicrobial peptides (AMPs) is an important strategy in innate immune. Processing of the precursor into the active form is a common posttranslational modification of AMPs in mammals. However, in invertebrates, the mechanism of AMP maturation is largely unknown. In the current study, to our knowledge, a novel potential AMP, designated as PcnAMP, was identified because of its significant induction by bacterial infection in the red swamp crayfish (Procambarus clarkii). PcnAMP was cleaved into a short fragment postinfection. Using the purified native peptide, this cleavage was found to be mediated by trypsin after synthesis. Proteolysis produced an N-terminal peptide that exerted the antibacterial function. Although the N-terminal peptide did not show significant similarity to any other sequences, it was predicted to have an overall helical structure and high amphipathicity, both of which are typical features of many AMPs. The N-terminal active peptide exhibited a wide spectrum of antimicrobial activity. Atomic force microscope imaging and flow cytometry analysis showed that treatment with the active form of PcnAMP led to the collapse of the bacterial cell wall and permeabilization of the bacterial cell membrane. Thus, this study provided a new candidate for therapeutic agent development, and revealed new insights into the maturation of AMPs in invertebrates.

The association between undiagnosed diabetes and cognitive function: findings from the China health and retirement longitudinal study.

BACKGROUND: The cognitive function of people with diabetes has gained an increasing interest in recent years, and this study focuses on exploring the relationship between undiagnosed diabetes and cognitive function among the middle-aged and elderly people in China. METHODS: The data came from the China Health and Retirement Longitudinal Study (CHARLS) which was conducted between July and October 2015. 9855 subjects were enrolled in the study. Executive function and episodic memory were used to assess cognitive function. The subjects were divided into three groups: no diabetes, diagnosed diabetes, and undiagnosed diabetes, and weighted multiple linear regression models were established to evaluate the association of undiagnosed diabetes with cognitive function. RESULTS: After controlling for covariates, undiagnosed diabetes was statistically associated with executive function (ß = -0.215, P < 0.01). In the age group of ≥65 years, undiagnosed diabetes was statistically associated with executive function (ß = -0.358, P < 0.01) and episodic memory (ß = -0.356, P < 0.01). When adjusting for confounders, no statistically significant associations were found between diagnosed diabetes and cognitive function except in 45-54 age group (ß = 0.374, P < 0.05). CONCLUSIONS: The cross-sectional study suggested that undiagnosed diabetes was linked to poor cognitive function, especially in the elderly population. Timely diagnosis and active treatment of diabetes are important to reduce the occurrence of cognitive impairment. Further prospective cohort studies are required to articulate the association between undiagnosed diabetes and cognitive function.

Does electroacupuncture benefit mixed urinary incontinence? A systematic review and meta-analysis with trial sequential analysis.

INTRODUCTIN AND HYPOTHESIS: Mixed urinary incontinence (MUI) comprises a combination of urgency and stress. The efficacy and safety of electroacupuncture (EA) for the treatment of MUI remain unclear. OBJECTIVE: To assess the efficacy and safety of EA in treating MUI. METHODS: We searched PubMed, CENTRAL, Embase, Web of Science, four Chinese databases, clinical research registration platforms, grey literature, and the reference lists of the selected studies. Risk of bias and quality were evaluated using the Revman 5.4 and Jadad scores. Meta-analysis was performed using Stata 15.1 software. Trial sequential analysis (TSA) was used to assess the stability of the results. RESULTS: Eight randomized controlled trials comprising 847 patients were included. The meta-analysis results showed that compared with antimuscarinic drugs plus pelvic floor muscle training, EA resulted in significantly less pad weight on the 1-h pad test and statistically significantly lower severity scores on the International Consultation on Incontinence Questionnaire Short Form. The change in the 72-h incontinence episode frequency difference was not statistically significant, and there was no outcome of overall response rate and quality of life in this meta-analysis. Few adverse events occurred in the EA group. The TSA results suggested that the result of change from baseline in the 1-h pad test was stable and the evidence was conclusive. CONCLUSIONS: EA could be a potential treatment option for MUI and is relatively safe. Nevertheless, because of the limitations of this study, our conclusions should be interpreted with caution, and further studies are needed to confirm the comprehensive clinical efficacy and placebo effect of EA.

Manipulating the Water Dissociation Electrocatalytic Sites of Bimetallic Nickel-Based Alloys for Highly Efficient Alkaline Hydrogen Evolution.

Transition-metal alloys are currently drawing increasing attention as promising electrocatalysts for the alkaline hydrogen evolution reaction (HER). However, traditional density-functional-theory-derived d-band theory fails to describe the hydrogen adsorption energy (ΔGH ) on hollow sites. Herein, by studying the ΔGH for a series of Ni-M (M=Ti, V, Cr, Mn, Fe, Co, Cu, Zn, Mo, W) bimetallic alloys, an improved d-band center was provided and a potential NiCu electrocatalyst with a near-optimal ΔGH was discovered. Moreover, oxygen atoms were introduced into Ni-M (O-NiM) to balance the adsorption/desorption of hydroxyl species. The tailored electrocatalytic sites for water dissociation can synergistically accelerate the multi-step alkaline HER. The prepared O-NiCu shows the optimum HER activity with a low overpotential of 23 mV at 10 mA cm-2 . This work not only broadens the applicability of d-band theory, but also provides crucial understanding for designing efficient HER electrocatalysts.