AHL-differential quorum sensing regulation of amino acid metabolism in Hafnia alvei H4.

Quorum sensing (QS) regulation of functional metabolites is rarely reported but a common trait of some bacteria. In this study, we found that QS promoted the extracellular accumulation of glycine and serine while inhibiting the extracellular accumulation of methionine in Hafnia alvei H4. The correlation analysis of five QS signals with the above three QS-regulated amino acids suggested that these QS signals may have functional differences in amino acid regulation. The exogenous AHL add-back studies on genes involved in glycine, serine, and methionine metabolic pathway highlighted that N-octanoyl-l-homoserine lactone (C8-HSL) downregulated the expression of sdhC/fumA genes involved in the succinate to malate pathway, thereby reducing the metabolic flux of the tricarboxylic acid (TCA) cycle as an amino acid metabolism platform. Further in-depth research revealed that the QS system promoted the conversion of folate to tetrahydrofolate (THF) by positively regulating the expression of folA and folM, thus impairing the ability of folate to promote methionine accumulation. Moreover, folate positively regulated the expression of the QS signal synthesis gene luxI, promoting the synthesis of QS signals, which may further enhance the influence of the QS system on amino acid metabolism. These findings contribute to the understanding of amino acid metabolism regulated by QS and provide new perspectives for accurate control of metabolic regulation caused by QS.IMPORTANCEAs one of the important regulatory mechanisms of microorganisms, quorum sensing (QS) is involved in the regulation of various physiological activities. However, few studies on the regulation of amino acid metabolism by QS are available. This study demonstrated that the LuxI-type QS system of Hafnia alvei H4 was involved in the regulation of multiple amino acid metabolism, and different types of QS signals exhibited different roles in regulating amino acid metabolism. Additionally, the regulatory effects of the QS system on amino acid metabolism were investigated from two important cycles that influence the conversion of amino acids, including the TCA cycle and the folate cycle. These findings provide new ideas on the role of QS system in the regulation of amino acid metabolism in organisms.

Potential quorum-sensing inhibitor of Hafnia alvei H4-theaflavin-3,3´-digallate analyzed by virtual screening and molecular simulation.

Hafnia species can cause food spoilage via the quorum-sensing (QS) system. Thus, strategies that target QS in these bacteria might be a good approach to safeguard the quality of processed food. In this study, the amino acid sequence of the LasI Ha protein, a key QS regulator from Hafnia alvei H4, was used to construct its 3D structure for the virtual screening of potential QS inhibitors (QSIs) from the Bioactive Compound database. Four potential QSIs were obtained, and these were all theaflavins (TFs). Among them, theaflavin-3,3´-digallate (TF3) was found to outperform the others, displaying a higher docking score according to molecular docking analysis, and required only a sub-minimal inhibitory concentration (31.25 mM) to cause a significant decrease in the production of the autoinducer N-acyl homoserine lactone in H. alvei H4 and up to 60.5% inhibition of its motility. Furthermore, molecular simulation results indicated that TF3 could stably bind to a cavity within LasI Ha to form stable hydrogen bonds and hydrophobic interactions with various key residues of the protein to exert the inhibitory effect. Thus, TF3 may be considered a potential compound to protect against food spoilage caused by H. alvei H4 via the quorum quenching. IMPORTANCE Hafnia alvei, the main strain studied in this paper, is often isolated from spoiled foods, especially refrigerated protein-based raw foods, and is generally considered to be a spoilage bacterium whose spoilage-causing properties may be closely related to its own very strong population-sensing activity, so the strategy of quorum quenching against H. alvei H4 may be a good way to guarantee the quality of processed foods. Given the current global requirements for food safety and quality, coupled with negative consumer perceptions of the excessive inclusion of synthetic chemicals in food products, the use of natural compounds as QSIs in the storage of aquatic food products would seem more attractive.

Dynamics of Bacterial Composition and Association with Quality Formation and Biogenic Amines Accumulation during Fish Sauce Spontaneous Fermentation.

High-throughput sequencing and high-pressure liquid chromatography (HPLC) methods were used to investigate the influences of microbial dynamics on the quality and biogenic amine (BA) content during fish sauce fermentation. The homogeneity of total viable bacteria and lactic acid bacteria in fish sauce becomes higher as fermentation progresses. Tetragenococcus was the key genus of fish sauce fermentation. Carnobacterium (38.43%) and Lentibacillus (41.01%) were the dominant genera in the samples fermented for 3 months and 18 months, respectively. These three bacterial genera were significantly related to the physicochemical characteristics and characteristic flavors of the sauces. Tetragenococcus was significantly positively correlated with nitrogen oxides, the main characteristic flavor components in fish sauce. The BA content in fish sauce fermentation increased from 106.88 to 376.03 mg/kg, and the content of histamine reached 115.30 mg/kg at the end of fermentation, indicating that fish sauce has health risks. About 66.67% of Lentibacillus isolates were able to produce a large amount of BA, suggesting that Lentibacillus was the key genus for BA accumulation in fish sauce fermentation. Research on reducing the content of BA in fish sauce by intervening with regard to the fermentation temperature showed that a safe fish sauce product could be obtained at the fermentation temperature of about 25°C. These results help us to understand the contribution of microbial community composition to fish sauce fermentation and provide a basis for improving the quality and safety of fermented fish sauce. IMPORTANCE Traditional fermentation of fish sauce is mainly carried out by complex microbial communities from raw anchovies and processing environments. However, it is still unclear how the environmental microbiota influences the quality and the safety of fish sauce products. Therefore, this study comprehensively explained the influence of microorganisms on the quality and safety of fish sauce during the fermentation process in terms of physicochemical characters, flavors, and BA. Additionally, the accumulation of BA in fish sauce fermentation was controlled by intervening in the fermentation temperature. This finding contributes to a deeper understanding of the role of environmental microbiota during fermentation and provides data support for improving the safety of fish sauce.

Quorum Sensing (QS)-regulated target predictions of Hafnia alvei H4 based on the joint application of genome and STRING database.

In this study, we sequenced and characterized the genome of H. alvei to grasp the genetic basis of its physiological activities, including QS, metabolism, virulence and antibiotic resistance, and then mapped these functional gene clusters obtained from KEGG pathways to the STRING database to predict the QS-regulated targets in these pathways. H. alvei was found to possess 63 QS-related genes, most of which were closely related to amino acid metabolism, especially methionine pathway, but were not directly related to carbon and energy metabolism. Furthermore, the adhesion gene clusters were closely relevant to the QS gene clusters as well as to the infection gene clusters, while only one node (KdsD) was predicted between the QS gene clusters and infection gene clusters, suggesting that QS might influence the infection by regulating adhesion. QS might confer cross-resistance to microorganisms not only by regulating the formation of biofilms but also by affecting the efflux of antibiotics. In addition, the interspecies and intraspecies patterns of absence/presence for QS and its target genes were determined to shed light on the conservation of the QS regulatory mechanism among the phylogenetically related species. Taken together, the proposed methodology could expand the spectrum of possible applications of genome-based analysis to decipher the basic metabolic relationship of a microorganism, especially when studying new isolates.

Efficacy and safety of radiofrequency in the treatment of trigeminal neuralgia: a systematic review and meta-analysis.

To assess the effectiveness of radiofrequency (RF) versus other percutaneous strategies (balloon compression, glycerol rhizotomy [GR], and microvascular decompression [MVD]) in patients with trigeminal neuralgia. We systematically searched the electronic databases of PubMed, Embase, and the Cochrane Library to identify eligible studies throughout October 2020. The odds ratio (OR) with 95% confidence interval (CI) was applied to assess effect estimates using the random-effects model. Eighteen retrospective cohort studies that enrolled 6391 patients with trigeminal neuralgia were included. We noted that RF was associated with an increased incidence of immediate pain relief compared with GR (OR 2.65; 95% CI 1.29-5.44; P = 0.008). Moreover, RF was associated with an increased risk of pain recurrence compared with MVD (OR 3.80; 95% CI 2.00-7.20; P < 0.001). Furthermore, RF was associated with an increased incidence of postoperative anesthesia compared with GR (OR 3.01; 95% CI 1.11-8.13; P = 0.030) or MVD (OR 4.62; 95% CI 2.15-9.93; P < 0.001). This study found that RF was superior to GR for the improvement in immediate pain relief; whereas, RF yielded an excess risk of pain recurrence compared with MVD. Moreover, the incidence of postoperative anesthesia in patients treated with RF significantly increased compared with the incidence after treatment with GR and MVD.

Quorum Sensing-Mediated and Growth Phase-Dependent Regulation of Metabolic Pathways in Hafnia alvei H4.

Quorum sensing (QS) is a widespread regulatory mechanism in bacteria used to coordinate target gene expression with cell density. Thus far, little is known about the regulatory relationship between QS and cell density in terms of metabolic pathways in Hafnia alvei H4. In this study, transcriptomics analysis was performed under two conditions to address this question. The comparative transcriptome of H. alvei H4 wild-type at high cell density (OD600 = 1.7) relative to low cell density (OD600 = 0.3) was considered as growth phase-dependent manner (GPDM), and the transcriptome profile of luxI/R deletion mutant (ΔluxIR) compared to the wild-type was considered as QS-mediated regulation. In all, we identified 206 differentially expressed genes (DEGs) mainly presented in chemotaxis, TCA cycle, two-component system, ABC transporters and pyruvate metabolism, co-regulated by the both density-dependent regulation, and the results were validated by qPCR and swimming phenotypic assays. Aside from the co-regulated DEGs, we also found that 59 DEGs, mediated by density-independent QS, function in pentose phosphate and histidine metabolism and that 2084 cell-density-dependent DEGs involved in glycolysis/gluconeogenesis and phenylalanine metabolism were influenced only by GPDM from significantly enriched analysis of transcriptome data. The findings provided new information about the interplay between two density-dependent metabolic regulation, which could assist with the formulation of control strategies for this opportunistic pathogen, especially at high cell density.

Radiofrequency Thermocoagulation Through Foramen Rotundum Versus Foramen Ovale for the Treatment of V2 Trigeminal Neuralgia.

BACKGROUND: Percutaneous radiofrequency thermocoagulation through the foramen rotundum (FR) is a new approach for the treatment of V2 trigeminal neuralgia (TN). OBJECTIVES: This study aimed to compare the efficacy and safety of the FR approach with that of the foramen ovale (FO) approach. STUDY DESIGN: Nonrandomized controlled clinical trial. SETTING: The study was conducted at Huaian Hospital of Huaian City, Huaian, China. METHODS: From July 2014 to December 2016, 80 consecutive patients with V2 TN were prospectively assigned into the FO group (n = 40) or the FR group (n = 40). All radiofrequency thermocoagulation procedures were performed under the guidance of digital subtraction angiography (DSA). Patients in the FO group were treated with Gasserian ganglion ablation through the Hartel approach. Patients in the FR group received ablation of the maxillary nerve at the internal opening of the FR. Facial pain was evaluated using the Visual Analog Scale preoperatively and postoperatively at 1 week, 6 months, and 1 year. RESULTS: All surgical procedures were successfully completed using DSA guidance. The FR group had no facial pain at postoperative 1 week, 6 months, and 1 year. The facial fain was not relieved in 4 patients of the FO group. They were treated with radiofrequency thermocoagulation of the maxillary nerve through the FR and maintained painless at postoperative 1 week, 6 months, and 1 year. At postoperative 1 year, another 3 patients relapsed in the FO group. The incidences of facial numbness and swelling did not differ significantly between the 2 groups (all P > 0.05). There was no postoperative corneal involvement or masticatory weakness in the FR group. However, corneal involvement and masticatory weakness occurred postoperatively in 22 (55%) patients and 31 (77.5%) patients in the FO group. The FR group had significantly shorter operation time than the FO group (19.3 ± 5.9 vs. 32.7 ± 8.7 minutes; P < 0.05). LIMITATIONS: We were unable to avoid the V1 and V3 branches, despite multiple adjustments of the needed position in 35 of the 40 patients in this group. CONCLUSIONS: For the treatment of V2 TN, thermocoagulation of the maxillary nerve through the FR had better efficacy and fewer complications in comparison with the Gasserian ganglion ablation through the FO. KEY WORDS: Neuralgia, pain, radiology, facial pain.

Coordination polymer hybridized Au nanocages: a nanoplatform for dual-modality imaging guided near-infrared driven photothermal therapy in vivo.

Coordination polymer hybridized Au nanocages (AuNC@CPs) were prepared via a dialysis process by combining Au nanocages with a coordination polymer, which was derived from (pq)2Ir(Hdcbpy) and Dy(OOCCH3)3. After hybridization, the localized surface plasmon resonance (LSPR) peak of the AuNC@CPs exhibited an evident red shift in comparison with that of the Au nanocages, which was due to a variation in the dielectric constant caused by the coordination polymer. Notably, inspired by their excellent photothermal, photoacoustic, and magnetic properties in solution, AuNC@CPs were used for near-infrared (NIR)-driven photothermal therapy (PTT) guided by photoacoustic (PA) and magnetic resonance (MR) imaging in vivo. Owing to their remarkable performance, the AuNC@CPs would represent promising nanoprobes for theranostics.