Salmonella manipulates macrophage migration via SteC-mediated myosin light chain activation to penetrate the gut-vascular barrier.

The intestinal pathogen Salmonella enterica rapidly enters the bloodstream after the invasion of intestinal epithelial cells, but how Salmonella breaks through the gut-vascular barrier is largely unknown. Here, we report that Salmonella enters the bloodstream through intestinal CX3CR1+ macrophages during early infection. Mechanistically, Salmonella induces the migration/invasion properties of macrophages in a manner dependent on host cell actin and on the pathogen effector SteC. SteC recruits host myosin light chain protein Myl12a and phosphorylates its Ser19 and Thr20 residues. Myl12a phosphorylation results in actin rearrangement, and enhanced migration and invasion of macrophages. SteC is able to utilize a wide range of NTPs other than ATP to phosphorylate Myl12a. We further solved the crystal structure of SteC, which suggests an atypical dimerization-mediated catalytic mechanism. Finally, in vivo data show that SteC-mediated cytoskeleton manipulation is crucial for Salmonella breaching the gut vascular barrier and spreading to target organs.

Bright Luminescence of Free Radical TEMPO Enabled by Electrochemiluminescence Technique.

Radicals can feature theoretically 100% light utilization owing to their nonelectron spin-forbidden transition and represent the most advanced luminescent materials at present. 2,2,6,6-Tetramethyl-1-piperidinyloxy (TEMPO) acts as a typically stable radical with very broad applications. However, their luminescent properties have not been discovered to date. In the present work, we observed the bright electrochemiluminescence (ECL) emission of TEMPO with a higher efficiency (72.3%) via the electrochemistry and coreactant strategies for the first time. Moreover, the radical-based ECL achieved high detection toward boron acid with a lower limit of detection (LOD) of 1.9 nM. This study offers a new approach to generate emissions for some unconventional luminophores and makes a major breakthrough in the field of new luminescent materials as well.

Hydrolysis mechanism of the cyclohexaborate anion: Unraveling the intricacies.

B 6 O 7 OH 6 2 - is a highly polymerized borate anion of three six-membered rings. Limited research on the B 6 O 7 OH 6 2 - hydrolysis mechanism under neutral solution conditions exists. Calculations based on density functional theory show that B 6 O 7 OH 6 2 - undergoes five steps of hydrolysis to form H3BO3 and B OH 4 - . At the same time, there are a small number of borate ions with different degrees of polymerization during the hydrolysis process, such as triborate, tetraborate, and pentaborate anions. The structure of the borate anion and the coordination environment of the bridging oxygen atoms control the hydrolysis process. Finally, this work explains that in existing experimental studies, the reason for the low B 6 O 7 OH 6 2 - content in solution environments with low total boron concentrations is that it depolymerizes into other types of borate ions and clarifies the borate species. The conversion relationship provides a basis for identifying the possibility of various borate ions existing in the solution. This work also provides a certain degree of theoretical support for the cause of the "dilution to salt" phenomenon.

Spatial transcriptomics reveals heterogeneity of macrophages in the tumor microenvironment of granulomatous slack skin.

Granulomatous slack skin (GSS) is an extremely rare subtype of cutaneous T-cell lymphoma accompanied by an abundant number of macrophages and is clinically characterized by the development of pendulous skin folds. However, the characteristics of these macrophages in GSS remain unclear. Here, we conducted a spatial transcriptomic study on one frozen GSS sample and drew transcriptomic maps of GSS for the first time. Gene expression analysis revealed the enrichment of three clusters with macrophage transcripts, each exhibiting distinct characteristics suggesting that their primary composition consists of different subpopulations of macrophages. The CD163+ /CD206+ cluster showed a tumor-associated macrophage (TAM) M2-like phenotype and highly expressed ZFP36, CCL2, TNFAIP6, and KLF2, which are known to be involved in T-cell interaction and tumor progression. The APOC1+ /APOE+ cluster presented a non-M1 or -M2 phenotype and may be related to lipid metabolism. The CD11c+ /LYZ+ cluster exhibited an M1-like phenotype. Notably, these cells strongly expressed MMP9, MMP12, CHI3L1, CHIT1, COL1A1, TIMP1, and SPP1, which are responsible for extracellular matrix (ECM) degradation and tissue remodeling. This may partially explain the symptoms of cutaneous relaxation in GSS. Further immunohistochemistry on four GSS cases demonstrated that CD11c predominantly marked granulomas and multinucleated giant cells, whereas CD163 was mainly expressed on scattered macrophages, appearing as a mutually exclusive pattern. The expression pattern of MMP9 overlapped with that of CD11c, implying that CD11c+ macrophages may be a source of MMP9. Our data shed light on the characteristics of macrophages in the GSS microenvironment and provide a theoretical basis for the application of MMP9 inhibitors to prevent cutaneous relaxation of GSS. © 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Multidimensional Ultrasound Evaluation of Diastasis Recti Abdominis During Different Gestational Periods.

OBJECTIVE: The purpose of this study is to explore the application value of two-dimensional ultrasound and shear wave elastography (SWE) in the multidimensional evaluation of diastasis recti abdominis (DRA) during different gestational periods. METHODS: A cohort of 202 gravidas that were examined in our hospital between June 2021 and September 2022 were selected for the purpose of the study, which included 26 cases with <14 weeks of pregnancy, 36 cases in the 14th-27th week of pregnancy, 36 cases in the 28th-34th week of pregnancy, 32 cases in the 35th-38th week of pregnancy, 45 cases at 42 days postpartum, and 27 cases at 3 months postpartum. The inter-rectus distance (IRD) and the thickness in each gestational period were measured, and Young's modulus of the rectus abdominis at different gestational periods was measured using SWE by two sonographers. The differences in IRD, thickness, and elasticity characteristics during different periods, and the correlation between rectus abdominis elasticity and IRD, thickness, body mass index (BMI), neonatal weight, and delivery mode were analyzed and compared. The consistency of SWE parameters obtained by different sonographers was also compared. RESULTS: There were significant differences in IRD, thickness, and Young's modulus during different gestational periods (P = .000, P < .001, P < .001). Early postpartum IRD and Young's modulus did not restore to the level of early pregnancy (P < .001, P < .001), while the thickness of rectus abdominis was not significantly different from that of early pregnancy (P = .211). The Young's modulus of rectus abdominis was negatively correlated with the IRD (r = .515), positively correlated with the thickness of rectus abdominis (r = .408), and weakly negatively correlated with maternal BMI (r = -.296). There was no significant correlation with neonatal weight or delivery mode (P = .147, .648). The Bland-Altman plot showed that the two sonographers had good consistency in evaluating the elasticity of rectus abdominis by SWE. CONCLUSION: The multidimensional evaluation of DRA by ultrasound is feasible and IRD and Young's modulus can be used to evaluate the postpartum recovery of DRA. The combination of the two can objectively reflect the severity of DRA morphology and function.

PLC1 mediated Cycloastragenol-induced stomatal movement by regulating the production of NO in Arabidopsis thaliana.

BACKGROUND: Astragalus grows mainly in drought areas. Cycloastragenol (CAG) is a tetracyclic triterpenoid allelochemical extracted from traditional Chinese medicine Astragalus root. Phospholipase C (PLC) and Gα-submit of the heterotrimeric G-protein (GPA1) are involved in many biotic or abiotic stresses. Nitric oxide (NO) is a crucial gas signal molecule in plants. RESULTS: In this study, using the seedlings of Arabidopsis thaliana (A. thaliana), the results showed that low concentrations of CAG induced stomatal closure, and high concentrations inhibited stomatal closure. 30 µmol·L-1 CAG significantly increased the relative expression levels of PLC1 and GPA1 and the activities of PLC and GTP hydrolysis. The stomatal aperture of plc1, gpa1, and plc1/gpa1 was higher than that of WT under CAG treatment. CAG increased the fluorescence intensity of NO in guard cells. Exogenous application of c-PTIO to WT significantly induced stomatal aperture under CAG treatment. CAG significantly increased the relative expression levels of NIA1 and NOA1. Mutants of noa1, nia1, and nia2 showed that NO production was mainly from NOA1 and NIA1 by CAG treatment. The fluorescence intensity of NO in guard cells of plc1, gpa1, and plc1/gpa1 was lower than WT, indicating that PLC1 and GPA1 were involved in the NO production in guard cells. There was no significant difference in the gene expression of PLC1 in WT, nia1, and noa1 under CAG treatment. The gene expression levels of NIA1 and NOA1 in plc1, gpa1, and plc1/gpa1 were significantly lower than WT, indicating that PLC1 and GPA1 were positively regulating NO production by regulating the expression of NIA1 and NOA1 under CAG treatment. CONCLUSIONS: These results suggested that the NO accumulation was essential to induce stomatal closure under CAG treatment, and GPA1 and PLC1 acted upstream of NO.

2-Heptanone, 2-nonanone, and 2-undecanone confer oxidation off-flavor in cow milk storage.

Flavor sensation is one of the most prevalent characteristics of food industries and an important consumer preference regulator of dairy products. So far, many volatile compounds have been identified, and their molecular mechanisms conferring overall flavor formation have been reported extensively. However, little is known about the critical flavor compound of a specific sensory experience in terms of oxidized off-flavor perception. Therefore, the present study aimed to compare the variation in sensory qualities and volatile flavors in full-fat UHT milk (FFM) and low-fat UHT milk (LFM) samples under different natural storage conditions (0, 4, 18, 25, 30, or 37°C for 15 and 30 d) and determine the main component causing flavor deterioration in the FFM and LFM samples using sensory evaluation, electronic nose, and headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). In addition, the Pearson correlation between the volatile flavor components and oxidative off-flavors was analyzed and validated by sensory reconstitution studies. Compared with the LFM samples, the FFM samples showed a higher degree of quality deterioration with increased storage temperature. Methyl ketones of odd carbon chains (i.e., 2-heptanone, 2-nonanone, 2-undecanone, 2-tridecanone, and 2-pentadecanone) reached a maximum content in the FFM37 samples over 30 d storage. The combined results of the Pearson correlation and sensory recombination study indicated that 2-heptanone, 2-nonanone, and 2-undecanone conferred off-flavor perception. Overall, the present study results provide potential target components for detecting and developing high-quality dairy products and lay a foundation for specific sensory flavor compound exploration in the food industry.

Bio-Inspired Polyanionic Electrolytes for Highly Stable Zinc-Ion Batteries.

For zinc-ion batteries (ZIBs), the non-uniform Zn plating/stripping results in a high polarization and low Coulombic efficiency (CE), hindering the large-scale application of ZIBs. Here, inspired by biomass seaweed plants, an anionic polyelectrolyte alginate acid (SA) was used to initiate the in situ formation of the high-performance solid electrolyte interphase (SEI) layer on the Zn anode. Attribute to the anionic groups of -COO- , the affinity of Zn2+ ions to alginate acid induces a well-aligned accelerating channel for uniform plating. This SEI regulates the desolvation structure of Zn2+ and facilitates the formation of compact Zn (002) crystal planes. Even under high depth of discharge conditions (DOD), the SA-coated Zn anode still maintains a stable Zn stripping/plating behavior with a low potential difference (0.114 V). According to the classical nucleation theory, the nucleation energy for SA-coated Zn is 97 % less than that of bare Zn, resulting in a faster nucleation rate. The Zn||Cu cell assembled with the SA-coated electrode exhibits an outstanding average CE of 99.8 % over 1,400 cycles. The design is successfully demonstrated in pouch cells, where the SA-coated Zn exhibits capacity retention of 96.9 % compared to 59.1 % for bare Zn anode, even under the high cathode mass loading (>10 mg/cm2 ).

Grafting of Tetraphenylethylene on Silica Surface, Characterizations, and Their Chromatographic Performance as Reversed-Phase Stationary Phases.

Surface modification is an effective way to functionalize the materials so as to get some special properties. Tetraphenylethylene (TPE) has been widely investigated as a well-known reagent which has the nature of aggregation-induced emission (AIE), but has never been reported in the liquid chromatography stationary phase. In this work, TPE-grafted silica (Sil-TPE) was obtained successfully using the derivative of 1-(4-hydroxyphenyl)-1,2,2-triphenylethylene as a ligand, and then characterized by elemental analysis, Fourier transform infrared spectra, thermogravimetric analysis, and so forth. Laser scanning confocal microscopy images reflected the AIE phenomenon of grafted TPE because the internal vibration and rotation of TPE molecules were restrained in the confined silica space. The contact angle test showed superhydrophobic properties of Sil-TPE. In order to understand thoroughly the mechanism of chromatographic performance and retention behavior for Sil-TPE, Tanaka test mixture, alkylbenzenes, polycyclic aromatic hydrocarbons (PAHs), and phenols were separated. This reveals that Sil-TPE has strong aromaticity and certain shape selectivity, especially, has excellent separation performance for PAHs and phenols. The thermodynamic properties and repeatability of Sil-TPE were further studied, which showed the stability of Sil-TPE. This work shows that TPE can be successfully grafted on silica surface and it has the potential to be a new kind of promising stationary phases in the future.

A potential revolution in cancer treatment: A topical review of FLASH radiotherapy.

FLASH radiotherapy (RT) is a novel technique in which the ultrahigh dose rate (UHDR) (≥40 Gy/s) is delivered to the entire treatment volume. Recent outcomes of in vivo studies show that the UHDR RT has the potential to spare normal tissue without sacrificing tumor control. There is a growing interest in the application of FLASH RT, and the ultrahigh dose irradiation delivery has been achieved by a few experimental and modified linear accelerators. The underlying mechanism of FLASH effect is yet to be fully understood, but the oxygen depletion in normal tissue providing extra protection during FLASH irradiation is a hypothesis that attracts most attention currently. Monte Carlo simulation is playing an important role in FLASH, enabling the understanding of its dosimetry calculations and hardware design. More advanced Monte Carlo simulation tools are under development to fulfill the challenge of reproducing the radiolysis and radiobiology processes in FLASH irradiation. FLASH RT may become one of standard treatment modalities for tumor treatment in the future. This paper presents the history and status of FLASH RT studies with a focus on FLASH irradiation delivery modalities, underlying mechanism of FLASH effect, in vivo and vitro experiments, and simulation studies. Existing challenges and prospects of this novel technique are discussed in this manuscript.

A novel strategy combining Mini-CEX and OSCE to assess standardized training of professional postgraduates in department of prosthodontics.

OBJECTIVE: Mini clinical evaluation exercise (mini-CEX) and objective structured clinical examination (OSCE) are widely acknowledged as effective measures of resident standardization training (RST) in European and American countries. However, in China primary mini-CEX and OSCE forms are mainly limited in undergraduate clinical examination. Little knowledge is available regarding the validity and right way of mini-CEX /OSCE evaluation system in advanced dental clinical education so far. This study aimed to explore whether combination of mini-CEX and OSCE represents a global-dimension assessment for postgraduate clinical competence in RST. METHODS: Postgraduates who received RST from June 2017 to June 2019 were selected and evaluated by modified mini-CEX/OSCE scales. Each student received evaluations at least twice in the initial and final stages of training (tested every 4 months). A questionnaire was conducted to investigate the satisfaction with the arrangement of RST. RESULTS: Mini-CEX/OSCE test results indicated that postgraduates have significantly improved their comprehensive competence in RST projects in the department of prosthodontics (P < 0.05). Compared to other master of Stomatology students, postgraduates taking up prosthodontics master's degree have made more progresses through a training period of up to 1 year and four sessions of face-to-face feedback tutoring (P < 0.05). Survey results revealed high level of satisfaction on clinical practice evaluation. CONCLUSION: Modified mini-CEX/OSCE combined evaluation system is an effective and reliable assessment tool for clinical comprehensive ability in the RST of professional graduates and can fully highlight their respective advantages on the improvement of students' clinical competency, especially after several rounds of assessments.

Experimental validation of proton physics models of Geant4 for calculating stopping power ratio.

In this work, we conducted experiments to validate the proton physics models of Geant4 (version 10.6). The stopping power ratios (SPRs) of 11 inserts, such as acrylic, delrin, high density polyethylene, and polytetrafluoroethylene, etc, were measured using a superconducting synchrocyclotron that produces a scattering proton beam. The SPRs of the inserts were also calculated based on Geant4 simulation with six physics lists, i.e. QGSP_ FTFP_ BERT, QGSP_BIC_HP, QGSP_BIC, QGSP_FTFP_BERT, QSGP_BERT, and QBBC. The calculated SPRs were compared to the experimental SPRs, and relative per cent error was used to quantify the accuracy of the simulated SPRs of inserts. The comparison showed that the five physics lists generally agree well with the experimental SPRs with a relative difference of less than 1%. The lowest overall percentage error was observed for QGSP_FTFP_BERT and the highest overall percentage error was observed for QGSP_BIC_HP. The 0.1 mm range cut value consistently led to higher percentage error for all physics lists except for QGSP_BIC_HP and QBBC. Based on the validation, we recommend QGSP_BERT_HP physics list for proton dose calculation.

Exploring the effect of silicene monolayer on the structure and function of villin headpiece and amyloid fibrils by molecular dynamics simulations.

With the development of various nanomaterial expected to be used in biomedical fields, it is more important to evaluate and understand their potential effects on biological system. In this work, two proteins with different structure, Villin Headpiece (HP35) with α-helix structure and protofibrils Aß1-42 with five ß-strand chains, were selected and their interactions with silicene were studied by means of molecular dynamics (MD) simulation to reveal the potential effect of silicene on the structure and function of biomolecules. The obtained results indicated that silicene could rapidly attract HP35 and Aß1-42 fibrils onto the surface to form a stable binding. The adsorption strength was moderate and no significant structural distortion of HP35 and Aß1-42 fibrils was observed. Moreover, the strength of calculated the H-bonds in neighbor chain of Aß1-42 fibrils indicated that the mild interactions between silicene and fibrils could regularize the structure of Aß1-42 fibrils and stabilize the interactions between five chains of fibrils protein, which might enhance the aggregation of Aß1-42 fibrils. This study provides a new insight for understanding the interaction between nanomaterials and biomolecules and moves forward the development of silicene into biomedical fields.

Osmotic stress-triggered stomatal closure requires Phospholipase Dδ and hydrogen sulfide in Arabidopsis thaliana.

Osmotic stress is one of the main stresses seriously affects the growth and development of plants. Hydrogen sulfide (H2S) emerges as the third gaseous signal molecule to involve in the complex network of signaling events. Phospholipase Dδ (PLDδ), as signal enzyme, responds to many biotic or abiotic stress responses. In this study, the functions and the relationship of PLDδ and H2S in stomatal closure induced by osmotic stress were explored. Using the seedlings of ecotype (WT), PLDδ deficient mutant (pldδ), L-cysteine desulfhydrase (LCD) deficient mutant (lcd) and pldδlcd double mutant as materials, the Real-time quantitative PCR (RT-qPCR) and the stomatal aperture were analyzed. Osmotic stress induced the expressions of PLDδ and LCD. The H2S content and the activities of PLD and LCD ascended in WT under osmotic stress. The phenotypes of pldδ, lcd and pldδlcd were more sensitive to osmotic stress than WT. Compared with pldδ, the stomatal of lcd showed lower sensitivity to osmotic stress, and the stomatal aperture of pldδlcd was similar to that of lcd. Simultaneous application of PA and NaHS resulted in tighter closure of stomatal than application of either PA or NaHS alone. These results suggested that osmotic stress-triggered stomatal closure requires PLDδ and H2S in A. thaliana. LCD acted downstream of PLDδ to regulate the stomatal closure induced by osmotic stress.

Adsorption Behaviors of Typical Proteins on BP, GR, and C2N Surfaces.

The biotoxicity of nanomaterials is very important for the application of nanomaterials in biomedical systems. In this study, proteins with varying secondary structures (α-helices, ß-sheets, and mixed α/ß structures) were employed to investigate the biological properties of three representative two-dimensional (2D) nanomaterials; these nanomaterials consisted of black phosphorus (BP), graphene (GR), and nitrogenized graphene (C2N) and were studied using molecular dynamics simulations. The results showed that the α-helix motif underwent a slight structural change on the BP surface and little structural change on the C2N surface. In contrast, the structure of the ß-sheet motif remained fairly intact on both the BP and C2N surfaces. The α-helix and ß-sheet motifs were able to freely migrate on the BP surface, but they were anchored to the C2N surface. In contrast to BP and C2N, GR severely disrupted the structures of the α-helix and ß-sheet motifs. BBA protein with mixed α/ß structures adsorbed on the BP and C2N surfaces and exhibited biological behaviors that were consistent with those of the α-helix and ß-sheet motifs. In summary, C2N may possess better biocompatibility than BP and GR and is expected to have applications in the biomedical field. This study not only comprehensively evaluated the biological characteristics of nanomaterials but also provided a theoretical strategy to explore and distinguish the surface characteristics of nanomaterials.

Array-based microbial identification upon extracellular aminoglycoside residue sensing.

Sensitive and rapid identification of pathogenic microorganisms is of great importance for clinical diagnosis and treatment. In this study, we developed an ultrasensitive colorimetric sensor array (CSA) based on the interactions between aminoglycoside antibiotics (AMGs) and Ag nanoparticles decorated with ß-cyclodextrin (AgNPs@ß-CD) to discriminate microorganisms quickly and accurately. Microorganisms can absorb different amounts of AMGs after incubation. Upon the addition of AgNPs@ß-CD, the corresponding extracellular AMG residues will bind to AgNPs@ß-CD, leading to color changes due to the modifications in localized surface plasmon resonance. The array was developed using 4 AMGs as sensing elements and AgNPs@ß-CD as the colorimetric probe to generate a unique colorimetric response pattern for each microorganism. Standard chemometric methods indicated excellent discrimination among 20 microorganisms at low concentrations of 2 × 106 CFU/mL. Therefore, this ultrasensitive CSA can be used for microbial discrimination portably and efficiently. Importantly, the concentration of microbial discrimination by our array is much lower than that of prior CSAs. This method of extracellular residue sensing also provided a new strategy to improve the sensitivity of conventional CSA in the discrimination of microorganisms, to measure the amount of intercellular uptake of AMGs by microorganisms, and to screen drugs that can easily be accumulated by the pathogenic microorganisms.

Dosimetric impact of range uncertainty in passive scattering proton therapy.

PURPOSE: The objective of this study was to investigate the dosimetric impact of range uncertainty in a large cohort of patients receiving passive scatter proton therapy. METHODS: A cohort of 120 patients were reviewed in this study retrospectively, of which 61 were brain, 39 lung, and 20 prostate patients. Range uncertainties of ±3.5% (overshooting and undershooting by 3.5%, respectively) were added and recalculated on the original plans, which had been planned according to our clinical planning protocol while keeping beamlines, apertures, compensators, and dose grids intact. Changes in the coverage on CTV and DVH for critical organs were compared and analyzed. Correlation between dose change and minimal distance between CTV and critical organs were also investigated. RESULTS: Although CTV coverages and maximum dose to critical organs were largely maintained for most brain patients, large variations over 5% were still observed sporadically. Critical organs, such as brainstem and chiasm, could still be affected by range uncertainty at 4 cm away from CTV. Coverage and OARs in lung and prostate patients were less likely to be affected by range uncertainty with very few exceptions. CONCLUSION: The margin recipe in modern TPS leads to clinically acceptable OAR doses in the presence of range uncertainties. However, range uncertainties still pose a noticeable challenge for small but critical serial organs near tumors, and occasionally for large parallel organs that are located distal to incident proton beams.

Tailoring Crystal Facets of Metal-Organic Layers to Enhance Photocatalytic Activity for CO2 Reduction.

It is common that different crystal facets in metal and metal oxide nanocrystals display different catalytic performances, whereas such phenomena have been rarely documented in metal-organic frameworks (MOFs). Herein, we demonstrate for the first time that a nickel metal-organic layer (MOL) exposing rich (100) crystal facets (Ni-MOL-100) shows a much higher photocatalytic CO2 -to-CO activity than the one exposing rich (010) crystal facets (Ni-MOL-010) and its bulky counterpart (bulky Ni-MOF), with a catalytic activity up to 2.5 and 4.6 times more active than Ni-MOL-010 and bulky Ni-MOF, respectively. Theoretical studies reveal that the two coordinatively unsaturated NiII ions with a close distance of 3.50 Šon the surface of Ni-MOL-100 enables synergistic catalysis, leading to more favorable energetics in CO2 reduction than that of Ni-MOL-010.

Distinct Expression of the Two NO-Forming Nitrite Reductases in Thermus antranikianii DSM 12462T Improved Environmental Adaptability.

Hot spring ecosystems are analogous to some thermal environments on the early Earth and represent ideal models to understand life forms and element cycling on the early Earth. Denitrification, an important component of biogeochemical nitrogen cycle, is highly active in hot springs. Nitrite (NO2-) reduction to nitric oxide (NO) is the significant and rate-limiting pathway in denitrification and is catalyzed by two types of nitrite reductases, encoded by nirS and nirK genes. NirS and NirK were originally considered incompatible in most denitrifying organisms, although a few strains have been reported to possess both genes. Herein, we report the functional division of nirS and nirK in Thermus, a thermophilic genus widespread in thermal ecosystems. Transcriptional levels of nirS and nirK coexisting in Thermus antranikianii DSM 12462T were measured to assess the effects of nitrite, oxygen, and stimulation time. Thirty-nine Thermus strains were used to analyze the phylogeny and distribution of nirS and nirK; six representative strains were used to assess the denitrification phenotype. The results showed that both genes were actively transcribed and expressed independently in T. antranikianii DSM 12462T. Strains with both nirS and nirK had a wider range of nitrite adaptation and revealed nir-related physiological adaptations in Thermus: nirK facilitated adaptation to rapid changes and extended the adaptation range of nitrite under oxygen-limited conditions, while nirS expression was higher under oxic and relatively stable conditions.

Characterization of Carbonyl Disinfection By-Products During Ozonation, Chlorination, and Chloramination of Dissolved Organic Matters.

Carbonyl compounds are an important class of by-products that are generated in disinfection reactions. These chemicals are ingredients contributing to toxicology in the drinking water system, the compositions and structures of which are worthy of attention. In this study, a chemical derivatization method based on simultaneous light/heavy isotope labeling was established for general recognition of carbonyl compounds and carbonyl disinfection by-products (DBPs) as per the humic substance reference standard (Suwannee river fulvic acid II, SRFA) before and after ozonation, chlorination, and chloramination. Decomposition of macromolecular components into polar carbonyl species was observed to be the most prominent pathway in ozone treatment due to the efficient reactivity of ozone with phenols and alkoxy aromatic rings. As a result, alteration of molecular characteristics was noticed. For instance, ozone-induced carbonyl DBPs in the highly oxygenated compound classes (0.67 ≤ O/C ≤ 1.2, 0.6 < H/C ≤ 1.5) possessed higher O/C but contained less oxygen numbers and carbon numbers. Cl/Br-carbonyl-DBPs were identified after chlorination and chloramination, and I-carbonyl-DBPs were found in ozone and chloramine treatments. Several major halogenated carbonyl homologues were further recognized, including halogenated 4-oxobutenoic acid analogues, halogenated 2,5-dioxohex-3-enoic acid analogues, and halogenated 4-cyclopentene-1,3-diones analogues. These findings illustrate the presence of abundant carbonyl DBPs in water disinfection, and hence their impacts on human health deserve further investigation.