Metabolism of novel potential syntrophic acetate-oxidizing bacteria in thermophilic methanogenic chemostats.

Acetate is a major intermediate in the anaerobic digestion of organic waste to produce CH4. In methanogenic systems, acetate degradation is carried out by either acetoclastic methanogenesis or syntrophic degradation by acetate oxidizers and hydrogenotrophic methanogens. Due to challenges in the isolation of syntrophic acetate-oxidizing bacteria (SAOB), the diversity and metabolism of SAOB and the mechanisms of their interactions with methanogenic partners are not fully characterized. In this study, the in situ activity and metabolic characteristics of potential SAOB and their interactions with methanogens were elucidated through metagenomics and metatranscriptomics. In addition to the reported SAOB classified in the genera Tepidanaerobacter, Desulfotomaculum, and Thermodesulfovibrio, we identified a number of potential SAOB that are affiliated with Clostridia, Thermoanaerobacteraceae, Anaerolineae, and Gemmatimonadetes. The potential SAOB possessing the glycine-mediated acetate oxidation pathway dominates SAOB communities. Moreover, formate appeared to be the main product of the acetate degradation by the most active potential SAOB. We identified the methanogen partner of these potential SAOB in the acetate-fed chemostat as Methanosarcina thermophila. The dominated potential SAOB in each chemostat had similar metabolic characteristics, even though they were in different fatty-acid-fed chemostats. These novel syntrophic lineages are prevalent and may play critical roles in thermophilic methanogenic reactors. This study expands our understanding of the phylogenetic diversity and in situ biological functions of uncultured syntrophic acetate degraders and presents novel insights into how they interact with methanogens.IMPORTANCECombining reactor operation with omics provides insights into novel uncultured syntrophic acetate degraders and how they perform in thermophilic anaerobic digesters. This improves our understanding of syntrophic acetate degradation and contributes to the background knowledge necessary to better control and optimize anaerobic digestion processes.

Opposite evolution of pathogenicity driven by in vivo wzc and wcaJ mutations in ST11-KL64 carbapenem-resistant Klebsiella pneumoniae.

AIMS: To investigate the in vivo evolution of the mucoid-phenotype of ST11-KL64 carbapenem-resistant Klebsiella pneumoniae (CRKP) isolated from the same patients and gain insights into diverse evolution and biology of these strains. METHODS: Whole genome sequencing and bioinformatic analysis were used to determine the mutation involved in the mucoid phenotype of ST11-KL64 CRKP. Gene knockout, bacterial morphology and capsular polysaccharides (CPS) extraction were used to verify the role of wzc and wcaJ in the mucoid phenotypes. Antimicrobial susceptibility, growth assay, biofilm formation, host cell adhesion and virulence assay were used to investigate the pleiotropic role of CPS changes in ST11-KL64 CRKP strains. RESULTS: Mutation of wzc S682N led to hypermucoid phenotype, which had negative impact on bacterial fitness and resulted in reduced biofilm formation and epithelial cell adhesion; while enhanced resistance to macrophage phagocytosis and virulence. Mutations of wcaJ gene led to non-mucoid phenotype with increased biofilm formation and epithelial cell adhesion, but reduced resistance of macrophage phagocytosis and virulence. Using virulence gene knockout, we demonstrated that CPS, rather than the pLVPK-like virulence plasmid, has a greater effect on mucoid phenotypic changes. CPS could be used as a surrogate marker of virulence in ST11-KL64 CRKP strains. CONCLUSIONS: ST11-KL64 CRKP strains sacrifice certain advantages to develop pathogenicity by changing CPS with two opposite in vivo evolution strategies.

Genetic diversity of siderophores and hypermucoviscosity phenotype in Klebsiella pneumoniae.

Siderophore is a group of low molecular weight compounds with high affinity for ferric iron, facilitating bacterial iron consumption, especially when competing with the host. Genetic diversity of the siderophore-encoding loci largely contributes to the intricacy of siderophore production and regulation in K. pneumoniae. We analyzed the genetic diversity of three acquired siderophores (salmochelin, yersiniabactin and aerobactin) in 272 K. pneumoniae strains from China, and revealed that hypervirulent and multidrug-resistant lineages had distinguished predominant siderophore loci. Strains producing more types of siderophores tend to present higher mucoviscosity levels, suggesting that siderophore production might synergistically interact with hypermucoviscosity phenotype.

Wave-packet behaviors of the defocusing nonlinear Schrödinger equation based on the modified physics-informed neural networks.

Investigated in this paper is the defocusing nonlinear Schrödinger (NLS) equation, which is used for describing the wave-packet dynamics in certain weakly nonlinear media. With the physics-informed neural networks (PINNs), we modify the corresponding loss function in the existing literature and obtain two types of dark solitons, type-I and type-II solitons. It is demonstrated that the modified loss function presents higher-precision wave-packet behaviors based on fewer initial and boundary data. Taking type-I solitons into consideration, we find that when only a small fraction of initial and boundary data are given, the prediction accuracy of the wave packets will be increased one or two orders of magnitude at least if the modification term of the loss function is introduced. Furthermore, for the inverse problem, the modified loss function provides a better estimate of the nonlinear coefficient of the NLS equation based on fewer observed data of the wave packets. For type-II solitons, we compare the required data and predicted results of the PINNs with those of the conventional time-splitting finite difference (TSFD) method and reveal that achieving the same precision of the wave-packet behavior, the PINNs with the modified loss functions require only one tenth of the amount of the initial and boundary data of the TSFD method. Besides, both unmodified and modified loss functions are exploited for predicting the behaviors of Gaussian wave packets, and it is observed that the predicted result of the modified loss function agrees with the high-precision solution of the time-splitting Fourier pseudospectral method, whereas the unmodified loss function fails.

Core Genome Allelic Profiles of Clinical Klebsiella pneumoniae Strains Using a Random Forest Algorithm Based on Multilocus Sequence Typing Scheme for Hypervirulence Analysis.

BACKGROUND: Hypervirulent Klebsiella pneumoniae (hvKP) infections can have high morbidity and mortality rates owing to their invasiveness and virulence. However, there are no effective tools or biomarkers to discriminate between hvKP and nonhypervirulent K. pneumoniae (nhvKP) strains. We aimed to use a random forest algorithm to predict hvKP based on core-genome data. METHODS: In total, 272 K. pneumoniae strains were collected from 20 tertiary hospitals in China and divided into hvKP and nhvKP groups according to clinical criteria. Clinical data comparisons, whole-genome sequencing, virulence profile analysis, and core genome multilocus sequence typing (cgMLST) were performed. We then established a random forest predictive model based on the cgMLST scheme to prospectively identify hvKP. The random forest is an ensemble learning method that generates multiple decision trees during the training process and each decision tree will output its own prediction results corresponding to the input. The predictive ability of the model was assessed by means of area under the receiver operating characteristic curve. RESULTS: Patients in the hvKP group were younger than those in the nhvKP group (median age, 58.0 and 68.0 years, respectively; P < .001). More patients in the hvKP group had underlying diabetes mellitus (43.1% vs 20.1%; P < .001). Clinically, carbapenem-resistant K. pneumoniae was less common in the hvKP group (4.1% vs 63.8%; P < .001), whereas the K1/K2 serotype, sequence type (ST) 23, and positive string tests were significantly higher in the hvKP group. A cgMLST-based minimal spanning tree revealed that hvKP strains were scattered sporadically within nhvKP clusters. ST23 showed greater genome diversification than did ST11, according to cgMLST-based allelic differences. Primary virulence factors (rmpA, iucA, positive string test result, and the presence of virulence plasmid pLVPK) were poor predictors of the hypervirulence phenotype. The random forest model based on the core genome allelic profile presented excellent predictive power, both in the training and validating sets (area under receiver operating characteristic curve, 0.987 and 0.999 in the training and validating sets, respectively). CONCLUSIONS: A random forest algorithm predictive model based on the core genome allelic profiles of K. pneumoniae was accurate to identify the hypervirulent isolates.

Prevalence and characteristics of pks gene cluster harbouring Klebsiella pneumoniae from bloodstream infection in China.

Bloodstream infection (BSI), caused by Klebsiella pneumoniae, is associated with high morbidity and mortality, where the pks gene cluster plays a major role in their occurrence and prevalence. Information on the prevalence and characteristics of this gene cluster in K. pneumoniae is currently limited in mainland China. We therefore undertook a multicentre longitudinal study which revealed the prevalence, overall, community-onset and hospital-acquired BSI to be 20.5%, 28.3% and 13.0%, respectively. Compared to pks-negative, pks-positive isolates were significantly more susceptible to antimicrobial agents with a low incidence (5.1%) of multidrug-resistance and with infrequent extended-spectrum beta-lactamase (ESBL) production. Among pks-positive isolates, ST23 (78/117) and ST65 (20/117) were the dominant sequence types, and the majority harboured virulence genes. Community-onset BSI patients infected with pks-positive isolates had a higher proportion of liver abscesses and a lower proportion of biliary obstructions (P < 0.05). The pks-positive isolates were mostly sporadic in the phylogenetic tree, with a 65.8 and 47.0 average allele difference between Clade 1 and Clade 2, respectively. We concluded that although pks-positive K. pneumoniae were generally susceptible to antimicrobials, the high prevalence of such isolates in community cases and the genotoxicity, merits further investigation.

Functional near-infrared spectroscopy in the evaluation of urban rail transit drivers' mental workload under simulated driving conditions.

The objective of this study is to investigate the potential of functional near-infrared spectroscopy (fNIRS) combined with heart rate variability indices, for the evaluation of the mental workload of urban rail transit drivers under simulated driving conditions, particularly during task engagement and disengagement. Experienced metro drivers wearing fNIRS monitoring systems were asked to drive for 90 min in a professional metro driving simulator. Workload stimulus tasks were added and an n-back task (n = 3) was implemented to induce workload in the simulated driving experiment. Experimental results indicate that fNIRS are sensitive to mental workload and reliable for discriminating the degree of mental workload. Research findings demonstrate the feasibility and reliability of fNIRS as a tool for real-time evaluating and monitoring driver mental workload along with task factors from a perspective of brain activations during simulated or actual driving. Practitioner Summary: This study provides evidence for the potential of functional near-infrared spectroscopy (fNIRS) for the evaluation of the mental workload of urban rail transit drivers under simulated driving conditions. The first fNIRS application to mental workload evaluation in the field of urban rail transportation helps companies develop reasonable shiftwork schedule and ensure operation safety. Abbreviations: fNIRS: functional near-infrared spectroscopy; oxy-Hb: Oxy-hemoglobin; NASA-TLX: National Aeronautics and Space Administration Task Load Index; EEG: electroencephalogram; ECG: electrocardiogram; HRV: variability; LF: low-frequency power; HF: high-frequency power; PFC: prefrontal cortex; NIRS: near-infrared spectroscopy; DWT: discrete wavelet transform; EMG: electromyography; DT: determination test; TP: total power; LFnorm: standardized LF; HFnorm: standardized HF; VLF: very low frequency; deoxy-Hb: deoxy-hemoglobin.

Molecular Epidemiology and Mechanism of Sulbactam Resistance in Acinetobacter baumannii Isolates with Diverse Genetic Backgrounds in China.

Sulbactam is a plausible option for treating Acinetobacter infections because of its intrinsic antibacterial activity against the members of the Acinetobacter genus, but the mechanisms of sulbactam resistance have not been fully studied in Acinetobacter baumannii In this study, a total of 2,197 clinical A. baumannii isolates were collected from 27 provinces in China. Eighty-eight isolates with various MICs for sulbactam were selected on the basis of their diverse clonality and underwent multilocus sequence typing (MLST), antimicrobial susceptibility testing, and resistance gene screening. The copy number and relative expression of blaTEM-1D and ampC were measured via quantitative PCR and quantitative reverse transcription-PCR, respectively. The genetic structure of multicopy blaTEM-1D was determined using the whole-genome sequencing technology. The cefoperazone-sulbactam resistance rate of the 2,197 isolates was 39.7%. The rate of positivity for blaTEM-1D or ISAba1-ampC in the sulbactam-nonsusceptible group (64.91% and 78.95%, respectively) was significantly higher than that in the sulbactam-susceptible group (0% and 0%, respectively; P < 0.001). The MIC of sulbactam (P < 0.001) varied considerably between the groups expressing ampC with or without upstream ISAba1 Notably, the genetic structure of the multicopy blaTEM-1D gene in strain ZS3 revealed that blaTEM-1D was embedded within four tandem copies of the cassette IS26-blaTEM-1D-Tn3-IS26 Therefore, blaTEM-1D and ISAba1-ampC represent the prevalent mechanism underlying sulbactam resistance in clinical A. baumannii isolates in China. The structure of the four tandem copies of blaTEM-1D first identified may increase sulbactam resistance.

Diversity of virulence level phenotype of hypervirulent Klebsiella pneumoniae from different sequence type lineage.

BACKGROUND: Hypervirulent Klebsiella pneumoniae (hvKP) is emerging around the Asian-Pacific region and it is the major cause of the community-acquired pyogenic liver abscesses. Multidrug-resistant hypervirulent Klebsiella pneumoniae (MDR-hvKP) isolates were reported in France, China and Taiwan. However, the international-ally agreed definition for hvKP and the virulence level of hvKP are not clear. RESULTS: In this study, 56 hvKP isolates were collected from March 2008 to June 2012 and investigated by string test, capsule serotyping, multilocus sequence typing (MLST), virulence gene detection and serum resistance assay. Among the 56 K. pneumoniae isolates, 64.3% had the hypermucoviscosity phenotype, meanwhile, 64.3% were the K1 serotype and 19.6% were the K2 serotype. Within the K1 serotype, 94.4% were ST23, and within the K2 serotype, ST65, ST86 and ST375 accounted for the same percentage 27.3%. The serum resistance showed statistically normal distribution. According to the 50% lethal dose of Galleria. mellonella infection model, hvKP isolates were divided into high virulence level group and moderate virulence level group. The ability of each method evaluating the virulence level of hvKP was assessed using the area under the receiver operating characteristic curve. CONCLUSIONS: K1 ST23 K. pneumoniae was the most prevalent clone of the hvKP. However, K1 ST23 K. pneumoniae was the dominant clone in the moderate virulence level group. MLST was a relatively reliable evaluation method to discriminate the virulence level of hvKP in our study.

Effect of Cd on growth, physiological response, Cd subcellular distribution and chemical forms of Koelreuteria paniculata.

Koelreuteria paniculata were cultivated in nutrient solution with different concentrations of Cd (0, 50, 150, 250 and 500 µM) and sampled after 90 days. The resistance, translocation, accumulation and stress responses in Koelreuteria paniculata were investigated by hydroponic experiments. The results showed that Koelreuteria paniculata is an efficient Cd excluder that can tolerate high concentrations of Cd (up to 150-250 µM of Cd). The concentration of Cd never exceeds 5 ppm in leaves and 10 ppm in roots. The high concentration of Cd (≥ 250 µM) had a toxic effect on K. paniculata and significantly restricted the plant growth. The accumulation ability of Cd by different plant tissues followed the sequence of roots > leaves > stems. The bioconcentration factors and translocation factors both were less than 1. Cd has the highest content in the cell wall and is migrated to soluble fractions and organelles at high concentrations. Undissolved Cd phosphate, pectates and protein-bound Cd were the predominant forms. The low concentration of Cd (≤150 µM) promoted the synthesis of soluble proteins, AsA and GSH, while high concentration of Cd clearly inhibited the physiological and biochemical process, caused membrane lipid peroxidation and severe membrane damages, and increased MDA and H2O2 contents. POD, CAT and SOD exhibited positive and effective responses to low concentration Cd stress, but could not remove the toxicity caused by high concentration Cd stress. The content of IAA, GA and ZT decreased and ABA content was significantly increased under high-concentration Cd stress.

Reduced Dimension Based Two-Dimensional DOA Estimation with Full DOFs for Generalized Co-Prime Planar Arrays.

In this paper, we investigate the problem of two-dimensional (2D) direction-of-arrival (DOA) estimation for generalized co-prime planar arrays. The classic multiple signal classification (MUSIC)-based methods can provide a superior estimation performance, but suffer from a tremendous computational burden caused by the 2D spectral search. To this end, we reduce the 2D problem into a one-dimensional (1D) one and propose a reduced dimension partial spectral search estimation method, which can compress the search region into a small 1D sector. Moreover, the proposed method can utilize the full information of the entire array without degrees-of-freedom loss. Furthermore, an iterative approach is also proposed to reduce complexity and improve performance. Simulation results show that the proposed methods can provide improved performance with substantially reduced complexity, as compared to other state-of-the-art methods.

A Low-Complexity ESPRIT-Based DOA Estimation Method for Co-Prime Linear Arrays.

The problem of direction-of-arrival (DOA) estimation is investigated for co-prime array, where the co-prime array consists of two uniform sparse linear subarrays with extended inter-element spacing. For each sparse subarray, true DOAs are mapped into several equivalent angles impinging on the traditional uniform linear array with half-wavelength spacing. Then, by applying the estimation of signal parameters via rotational invariance technique (ESPRIT), the equivalent DOAs are estimated, and the candidate DOAs are recovered according to the relationship among equivalent and true DOAs. Finally, the true DOAs are estimated by combining the results of the two subarrays. The proposed method achieves a better complexity-performance tradeoff as compared to other existing methods.

Application of shear wave elastography in fine needle aspiration biopsy for thyroid nodule.

Ultrasound guided fine needle aspiration biopsy (FNAB) is the first-line procedure in diagnosing thyroid nodules. However, emerging studies have reported its unsatisfactory cell sample for cytology evaluation. In this case report, shear wave elastography (SWE) guided FNAB on a thyroid nodule is presented. Biopsies were performed on both hard and soft areas of the same nodule on the SWE image. Cytological results demonstrated the sufficient diagnostic components and typical nuclear features of papillary carcinoma found on the specimen from the hard area. This case study indicates SWE is a useful complementary tool to conventional B-mode ultrasound in guiding thyroid nodule FNAB.

Clinical and laboratory insights into the threat of hypervirulent Klebsiella pneumoniae.

Hypervirulent Klebsiella pneumoniae (hvKP) typically causes severe invasive infections affecting multiple sites in healthy individuals. In the past, hvKP was characterized by a hypermucoviscosity phenotype, susceptibility to antimicrobial agents, and its tendency to cause invasive infections in healthy individuals within the community. However, there has been an alarming increase in reports of multidrug-resistant hvKP, particularly carbapenem-resistant strains, causing nosocomial infections in critically ill or immunocompromised patients. This presents a significant challenge for clinical treatment. Early identification of hvKP is crucial for timely infection control. Notably, identifying hvKP has become confusing due to its prevalence in nosocomial settings and the limited predictive specificity of the hypermucoviscosity phenotype. Novel virulence predictors for hvKP have been discovered through animal models or machine learning algorithms, while standardization of identification criteria is still necessary. Timely source control and antibiotic therapy have been widely employed for the treatment of hvKP infections. Additionally, phage therapy is a promising alternative approach due to escalating antibiotic resistance. In summary, this narrative review highlights the latest research progress in the development, virulence factors, identification, epidemiology of hvKP, and treatment options available for hvKP infection.

Development of a Novel Typing Scheme Based on the Genetic Diversity of Heme/Hemin Uptake System Hmu in Klebsiella pneumoniae Species Complex.

Iron is essential for the survival and reproduction of Klebsiella pneumoniae. Although K. pneumoniae employs multiple types of siderophores to scavenge iron during infections, the majority of host iron is retained within erythrocytes and carried by hemoglobin that is inaccessible to siderophores. HmuRSTUV is a bacterial hemin/hemoprotein uptake system. However, the genetic background and function of HmuRSTUV in K. pneumoniae remain unknown. We collected 2,242 K. pneumoniae genomes, of which 2,218 (98.9%) had complete hmuRSTUV loci. Based on the 2,218 complete hmuRSTUV sequences, we established a novel typing scheme of K. pneumoniae named hmST, and 446 nonrepetitive hmSTs were identified. In hypervirulent lineages, hmST was diversely distributed and hmST1 mainly existed in ST23 strains. In contrast, hmST was less diversely distributed among multidrug-resistant strains. hmST demonstrated greater genetic diversity in hypervirulent lineages and community-acquired and bloodstream-sourced strains. In vitro and in vivo experiments revealed that an intact hmuRSTUV was essential for hemin uptake, playing an important role in bloodstream infections. This study established a novel typing scheme of hmST based on hmuRSTUV providing new insights into identifying and monitoring the emergence of novel virulence evolution in K. pneumoniae. IMPORTANCE Siderophore is a group of low molecular weight compounds with high affinity for ferric iron, which could facilitate bacterial iron consumption. Similarly, hemin/heme scavenged by the hemin uptake system HmuRSTUV usually act as another critical iron source for K. pneumoniae. This study proved that Hmu system significantly promoted the growth of K. pneumoniae in the presence of hemin and played an important role in bloodstream infections. A novel typing scheme named hmST was established, and the genetic diversity of hmuRSTUV loci was analyzed based on a large number of genomes. This study provides new insights into identifying and monitoring the emergence of novel virulence evolution in K. pneumoniae.

Metalloporphyrin hemin modified carbon nanotube decorated titanium carbide with redox catalytic ability for electrochemical determination of hydrogen peroxide and uric acid.

Herein, a functional hybrid named metalloporphyrin hemin modified carbon nanotube decorated titanium carbide (Hemin/CNT/Ti3C2Tx) with redox catalytic ability is studied and applied to electrochemical sensing. The morphology, crystal structure and chemical composition of the hybrid are investigated through field emission scanning electron microscope (FESEM), transmission electron microscope (TEM), X-ray diffractometer (XRD), X-ray photoelectron spectroscopy (XPS), thermogravimetric analysis (TGA), and Brunauer-Emmett-Teller (BET) analysis. Benefiting from the unique multilayer structure, strong enrichment ability, excellent electrochemical performance of CNT/Ti3C2Tx and outstanding catalytic property of hemin, the developed Hemin/CNT/Ti3C2Tx hybrid presents desirable performance for electrochemical sensing. Two independent electrochemical sensing techniques including chronoamperometry (i-t) and differential pulse voltammetry (DPV) are adopted to realize hydrogen peroxide (H2O2) and uric acid (UA) determination, demonstrating the excellent redox catalytic ability of Hemin/CNT/Ti3C2Tx. More importantly, it is successfully applied to real-time monitoring of H2O2 released from living cells, and accurate determination of UA in urine samples is also realized, demonstrating a good practicability.