Improving Interface Matching in MOF-on-MOF S-Scheme Heterojunction through π-π Conjugation for Boosting Photoelectric Response.

Accelerating the migration of interfacial carriers in a heterojunction is of paramount importance for driving high-performance photoelectric responses. However, the inferior contact area and large resistance at the interface limit the eventual photoelectric performance. Herein, we fabricated an S-scheme heterojunction involving a 2D/2D dual-metalloporphyrin metal-organic framework with metal-center-regulated CuTCPP(Cu)/CuTCPP(Fe) through electrostatic self-assembly. The ultrathin nanosheet-like architectures reduce the carrier migration distance, while the similar porphyrin backbones promote reasonable interface matching through π-π conjugation, thereby inhibiting the recombination of photogenerated carriers. Furthermore, the metal-center-regulated S-scheme band alignments create a giant built-in electric field, which provides a huge driving force for efficient carrier separation and migration. Coupling with the biomimetic catalytic activity of CuTCPP(Fe), the resultant heterojunction was utilized to construct photoelectrochemical uric acid biosensors. This work provides a general strategy to enhance photoelectric responses by engineering the interfacial structure of heterojunctions.

Novel Wide-Working-Temperature NaNO3-KNO3-Na2SO4 Molten Salt for Solar Thermal Energy Storage.

A novel ternary eutectic salt, NaNO3-KNO3-Na2SO4 (TMS), was designed and prepared for thermal energy storage (TES) to address the issues of the narrow temperature range and low specific heat of solar salt molten salt. The thermo-physical properties of TMS-2, such as melting point, decomposition temperature, fusion enthalpy, density, viscosity, specific heat capacity and volumetric thermal energy storage capacity (ETES), were determined. Furthermore, a comparison of the thermo-physical properties between commercial solar salt and TMS-2 was carried out. TMS-2 had a melting point 6.5 °C lower and a decomposition temperature 38.93 °C higher than those of solar salt. The use temperature range of TMS molten salt was 45.43 °C larger than that of solar salt, which had been widened about 13.17%. Within the testing temperature range, the average specific heat capacity of TMS-2 (1.69 J·K-1·g-1) was 9.03% higher than that of solar salt (1.55 J·K-1·g-1). TMS-2 also showed higher density, slightly higher viscosity and higher ETES. XRD, FTIR and Raman spectra SEM showed that the composition and structure of the synthesized new molten salt were different, which explained the specific heat capacity increasing. Molecular dynamic (MD) simulation was performed to explore the different macroscopic properties of solar salt and TMS at the molecular level. The MD simulation results suggested that cation-cation and cation-anion interactions became weaker as the temperature increased and the randomness of molecular motion increased, which revealed that the interaction between the cation cluster and anion cluster became loose. The stronger interaction between Na-SO4 cation-anion clusters indicated that TMS-2 molten salt had a higher specific heat capacity than solar salt. The result of the thermal stability analysis indicated that the weight losses of solar salt and TMS-2 at 550 °C were only 27% and 53%, respectively. Both the simulation and experimental study indicated that TMS-2 is a promising candidate fluid for solar power generation systems.

Horseradish Peroxidase Immobilized in Metal Aerogels Boosts Electron Transfer and an Interfacial Reaction for Photoelectrochemical Immunoassay.

Photoelectrochemical (PEC) enzymatic biosensors have attracted widespread attention for their specificity and sensitivity, but the charge migration between an enzyme and a semiconductor remains uncertain. In this work, horseradish peroxidase (HRP) was successfully immobilized on ionic liquid-functioned Cu@Cu2O (IL-Cu@Cu2O) aerogels to boost charge transfer and an interfacial redox reaction. The photogenerated electrons flow from the conduction band of Cu2O to HRP under the assistance of Cu and are subsequently captured by [Fe(CN)6]3- in the electrolyte, which boosts the PEC response. The improved interfacial catalytic ability after the immobilization of HRP is proved by the enhanced redox ability under light irradiation. Benefiting from the excellent PEC activity and catalysis reaction of IL-Cu@Cu2O@HRP, an immunoassay platform was constructed for sensing prostate-specific antigens, which presents a wide detection range and a low limit of detection. An in-depth understanding of the direct electronic communication between a photoactive material and an enzyme for boosted charge transfer and interfacial catalysis provides a new view for the design of advanced PEC sensing platforms.

Yellow Carbon Dots for Fluorescent Water Sensing, Relative Humidity Sensing, and Anticounterfeiting Applications.

Most fluorescent probes based on carbon dots (CDs) fluorescence color or intensity change are still used for detection in solution, but in practical fluorescence detection applications, detection in the solid state is necessary. Therefore, a CDs-based fluorescence sensing device is designed in this paper, which can be used for water detection in liquid and solid states. Using oPD as a single precursor, yellow fluorescent CDs (y-CDs) were prepared by hydrothermal method, which can be used in the field of water detection and anti-counterfeiting by using its solvent-sensitive properties. First, y-CDs can be used to visually and intelligently detect the water content in ethanol. Secondly, it can be used to detect the Relative Humidity (RH) of the environment by combining it with cellulose to form a fluorescent film. Finally, y-CDs can also be used as a fluorescent material for fluorescence anti-counterfeiting.

Mechanism of METTL14 and m6A modification of lncRNA MALAT1 in the proliferation of oral squamous cell carcinoma cells.

OBJECTIVES: Methyltransferase-like 14 (METTL14) plays an epigenetic role in various cancer through N6-methyladenosine (m6A) modification. This study sought to analyze the mechanism of METTL14 in oral squamous cell carcinoma (OSCC) cell proliferation. METHODS: Expression levels of METTL14, lncRNA metastasis associated with lung adenocarcinoma transcript 1 (lncRNA MALAT1), microRNA (miR)-224-5p, and histone lysine demethylase 2A (KDM2A) in OSCC tissues (N = 40), and cell lines (FaDu, SCC-25, CAL-27, and SCC-15) were detected. Cell viability and colony formation capacity were assessed. m6A level, stability, and subcellular localization of lncRNA MALAT1 were determined. Nude mouse xenograft tumor assay was performed to confirm the role of METTL14 in vivo. RESULTS: METTL14 and lncRNA MALAT1 were upregulated, and miR-224-5p was downregulated in OSCC tissues and cells. Silencing METTL14 repressed OSCC cell viability and colony formation. Overexpression of MALAT1 and KDM2A or miR-224-5p downregulation reversed the inhibition of silencing METTL14 on OSCC cell proliferation. METTL14 induced m6A modification of MALAT1 to upregulate MALAT1. MALAT1 is comparatively bound to miR-224-5p to promote KDM2A transcription. In vivo, METTL14 promoted tumor growth via regulating MALAT1/miR-224-5p/ KDM2A. CONCLUSIONS: Overall, our findings verified the therapeutic role of silencing METTL14 in OSCC treatment through the MALAT1/miR-224-5p/KDM2A axis.

Chemically Stable Guanidinium Covalent Organic Framework for the Efficient Capture of Low-Concentration Iodine at High Temperatures.

The capture of radioactive I2 vapor from nuclear waste under industrial operating conditions remains a challenging task, as the practical industrial conditions of high temperature (≥150 °C) and low I2 concentration (∼150 ppmv) are unfavorable for I2 adsorption. We report a novel guanidinium-based covalent organic framework (COF), termed TGDM, which can efficiently capture I2 under industrial operating conditions. At 150 °C and 150 ppmv I2, TGDM exhibits an I2 uptake of ∼30 wt %, which is significantly higher than that of the industrial silver-based adsorbents such as Ag@MOR (17 wt %) currently used in the nuclear fuel reprocessing industry. Characterization and theoretical calculations indicate that among the multiple types of adsorption sites in TGDM, only ionic sites can bond to I2 through strong Coulomb interactions under harsh conditions. The abundant ionic groups of TGDM account for its superior I2 capture performance compared to various benchmark adsorbents. In addition, TGDM exhibits exceptionally high chemical and thermal stabilities that fully meet the requirements of practical radioactive I2 capture (high-temperature, humid, and acidic environment) and differentiate it from other ionic COFs. Furthermore, TGDM has excellent recyclability and low cost, which are unavailable for the current industrial silver-based adsorbents. These advantages make TGDM a promising candidate for capturing I2 vapor during nuclear fuel reprocessing. This strategy of incorporating chemically stable ionic guanidine moieties in COF would stimulate the development of new adsorbents for I2 capture and related applications.

Ultra-Low Content Bismuth-Anchored Gold Aerogels with Plasmon Property for Enhanced Nonenzymatic Electrochemical Glucose Sensing.

Effective glucose surveillance provides a strong guarantee for the high-quality development of human health. Au nanomaterials possess compelling applications in nonenzymatic electrochemical glucose biosensors owing to superior catalytic performances and intriguing biocompatibility properties. However, it has been a grand challenge to accurately control the architecture and composition of Au nanomaterials to optimize their optical, electronic, and magnetic properties for further improving the performance of electrocatalytic sensing. Herein, ultra-low content Bi-anchored Au aerogels are synthesized via a one-step reduction strategy. Benefiting from the unique structure of aerogels as well as the synergistic effect between Au and Bi, the optimized Au200Bi aerogels greatly boost the activity of glucose oxidation compared with Au aerogels. Under plasmon resonance excitation, bimetallic Au200Bi aerogels with wider photics-dependent properties further show plasmon-promoted glucose electro-oxidation activity, which is derived from the photothermal and photoelectric effects caused by the local surface plasmon resonance. Thanks to the enhanced performance, a nonenzymatic electrochemical glucose biosensor is constructed to detect glucose with high sensitivity. This plasmon-promoted electrocatalytic activity through the synergetic strategy of bimetallic aerogels has potential applications in various research fields.

Quantitative mapping of DNA phosphorothioatome reveals phosphorothioate heterogeneity of low modification frequency.

Phosphorothioate (PT) modifications of the DNA backbone, widespread in prokaryotes, are first identified in bacterial enteropathogens Escherichia coli B7A more than a decade ago. However, methods for high resolution mapping of PT modification level are still lacking. Here, we developed the PT-IC-seq technique, based on iodine-induced selective cleavage at PT sites and high-throughput next generation sequencing, as a mean to quantitatively characterizing the genomic landscape of PT modifications. Using PT-IC-seq we foud that most PT sites are partially modified at a lower PT frequency (< 5%) in E. coli B7A and Salmonella enterica serovar Cerro 87, and both show a heterogeneity pattern of PT modification similar to those of the typical methylation modification. Combining the iodine-induced cleavage and absolute quantification by droplet digital PCR, we developed the PT-IC-ddPCR technique to further measure the PT modification level. Consistent with the PT-IC-seq measurements, PT-IC-ddPCR analysis confirmed the lower PT frequency in E. coli B7A. Our study has demonstrated the heterogeneity of PT modification in the bacterial population and we also established general tools for rigorous mapping and characterization of PT modification events at whole genome level. We describe to our knowledge the first genome-wide quantitative characterization of PT landscape and provides appropriate strategies for further functional studies of PT modification.

MIR210HG is aberrantly expressed in the seminal plasma of varicocele patients and associated with varicocele-related dyszoospermia.

This study aimed to confirm the expression of the seminal plasma long noncoding RNAs (lncRNAs) microRNA210 host gene (MIR210HG) in varicocele (VC) patients, to further explore the association between MIR210HG and VC severity and to evaluate whether MIR210HG can predict VC-related dyszoospermia. Semen samples from 188 VC patients and 92 healthy men were collected. Quantitative reverse transcriptase PCR detected seminal plasma MIR210HG levels. Receiver operating characteristic analysis assessed the ability of MIR210HG to screen patients with VC, or to screen VC patients with abnormal semen quality. Logistic analysis assessed the value of MIR210HG in predicting dyszoospermia in VC patients. The levels of MIR210HG in seminal plasma of VC patients were upregulated, which could screen VC patients. In addition, the levels of seminal plasma MIR210HG were upregulated with VC severity and were downregulated at 6 months after surgery in VC patients. Moreover, elevated MIR210HG levels in VC patients with abnormal semen quality could screen patients with abnormal semen quality and could independently predict the occurrence of dyszoospermia in VC patients. Seminal plasma MIR210HG expression is upregulated in VC patients, is associated with the severity of VC and may function as an independent predictor of VC-related dyszoospermia.

Lesion Heterogeneity and Long-Term Heteroresistance in Multidrug-Resistant Tuberculosis.

Tuberculosis heteroresistance, in which only a fraction of the bacteria in a patient with tuberculosis contains drug-resistant mutations, has been a rising concern. However, its origins and prevalence remain elusive. Here, whole-genome sequencing was performed on 83 serial isolates from 31 patients with multidrug-resistant tuberculosis, and heteroresistance was detected in isolates from 21 patients (67.74%). Heteroresistance persisted in the host for long periods, spanning months to years, and was associated with having multiple tubercular lesions. Our findings indicate that heteroresistance is common and persistent in patients with multidrug-resistant tuberculosis and may affect the success of their treatment regimens.

Proton-Regulated Catalytic Activity of Nanozymes for Dual-Modal Bioassay of Urease Activity.

Benefiting from the merits of high stability and superior activity, nanozymes are recognized as promising alternatives to natural enzymes. Despite the great leaps in the field of therapy and colorimetric sensing, the development of highly sensitive nanozyme-involved photoelectrochemical (PEC) biosensors is still in its infancy. Specifically, the investigation of multifunctional nanozymes facilitating different catalytic reactions remains largely unexplored due to the difficulty in synergistically amplifying the PEC signals. In this work, mesoporous trimetallic AuPtPd nanospheres were synthesized with both efficient oxidase and peroxidase-like activities, which can synergistically catalyze the oxidation of 4-chloro-1-naphthol to produce benzo-4-chlorohexadienone precipitation on the surface of photoactive materials, and thus lead to the decreased photocurrent as well as increased charge-transfer resistance. Inspired by the proton-dependent catalytic activity of nanozymes, a self-regulated dual-modal PEC and electrochemical bioassay of urease activity was innovatively established by in situ regulating the activity of AuPtPd nanozymes through urease-mediated proton-consuming enzymatic reactions, which can remarkably improve the accuracy of the assay. Meanwhile, the determination of urease activity in spiked human saliva samples was successfully realized, indicating the reliability of the biosensor and its application prospects in clinical diagnosis.

Post-operative blood pressure and 3-year major adverse cardiac events in Chinese patients undergoing PCI.

BACKGROUND: There is no clear evidence for the target value of blood pressure control after Percutaneous coronary intervention (PCI). Therefore, our study was designed to explore the relationship between blood pressure after PCI and major adverse cardiac events (MACE) during 3-year follow-up. METHODS: This study is a prospective study. We included the patients who were diagnosed with acute coronary syndrome and underwent PCI stent implantation operation. The study initially collected information of 552 patients. The start and end times of the study are from January 1, 2017 to December 31, 2020. The independent variables of this study are the average systolic blood pressure and the average diastolic blood pressure after PCI. The dependent variable is the occurrence of MACE events in patients within 3 years after PCI. MACE is defined as acute myocardial infarction, recurring chest pain, heart failure, stroke, revascularization and cardiac death. RESULTS: A total of 514 subjects met the inclusion criteria. The average age of the study subjects is 61.92 ± 9.49 years old, of which 67.12% are male. 94 subjects had a MACE event within 3 years, and the occurrence rate was 18.29%. There is no significant non-linear or linear relationship between diastolic blood pressure and MACE events. There is a curvilinear relationship between the average systolic blood pressure of patients after PCI and MACE events within 3 years and the inflection point is 121. On the left side of the inflection point, the effect size and 95% CI are 1.09 and 1.01-1.18, respectively (P = 0.029). The impact size and 95% CI at the right inflection point were 1.00 and 0.98-1.02(P = 0.604), respectively. CONCLUSION: There is a curvilinear relationship between systolic blood pressure and prognosis of patients after PCI. Under the premise of ensuring the safety of patients, maintaining lower blood pressure after surgery is beneficial to improve the prognosis of patients.

Hormetic Effects of Dimethachlone on Mycelial Growth and Virulence of Sclerotinia sclerotiorum.

Fungicide hormesis has implications for the application of fungicides to control plant diseases. We investigated the hormetic effects of the dicarboximide fungicide dimethachlone on mycelial growth and virulence of the necrotrophic plant pathogen Sclerotinia sclerotiorum. Dimethachlone at sublethal doses in potato dextrose agar (PDA) increased the mycelial growth of S. sclerotiorum. After the growth-stimulated mycelia were subcultured on fresh PDA and inoculated on rapeseed leaves, increased mycelial growth and virulence were observed, indicating that hormetic traits were passed down to the next generation. Dimethachlone applied to leaves at 0.002 to 500 µg/ml stimulated virulence, with a maximum stimulation amplitude (MSA) of 31.4% for the isolate HLJ4, which occurred at 2 µg/ml. Dimethachlone-resistant isolates and transformants had a mean virulence MSA of 30.4%, which was significantly higher (P = 0.008) than the MSA for sensitive isolates (16.2%). Negative correlations were detected between MSA and virulence in the absence of any fungicide (r = -0.872, P < 0.001) and between MSA and mycelial growth on PDA (r = -0.794, P = 0.002). Studies on hormetic mechanisms indicated that dimethachlone had no significant effects on expression levels of three virulence-associated genes, that is, a cutinase-encoding gene SsCut, a polygalacturonase gene SsPG1, or an oxaloacetate acetylhydrolase gene SsOah1. The results will contribute to understanding hormesis and have implications for the judicious application of fungicides to control plant diseases.

Citywide Transmission of Multidrug-resistant Tuberculosis Under China's Rapid Urbanization: A Retrospective Population-based Genomic Spatial Epidemiological Study.

BACKGROUND: Population movement could extend multidrug-resistant tuberculosis (MDR-TB) transmission and complicate its global prevalence. We sought to identify the high-risk populations and geographic sites of MDR-TB transmission in Shenzhen, the most common destination for internal migrants in China. METHODS: We performed a population-based, retrospective study in patients diagnosed with MDR-TB in Shenzhen during 2013-2017. By defining genomic clusters with a threshold of 12-single-nucleotide polymorphism distance based on whole-genome sequencing of their clinical strains, the clustering rate was calculated to evaluate the level of recent transmission. Risk factors were identified by multivariable logistic regression. To further delineate the epidemiological links, we invited the genomic-clustered patients to an in-depth social network investigation. RESULTS: In total, 105 (25.2%) of the 417 enrolled patients with MDR-TB were grouped into 40 genome clusters, suggesting recent transmission of MDR strains. The adjusted risk for student to have a clustered strain was 4.05 (95% confidence interval, 1.06-17.0) times greater than other patients. The majority (70%, 28/40) of the genomic clusters involved patients who lived in different districts, with residences separated by an average of 8.76 kilometers. Other than household members, confirmed epidemiological links were also identified among classmates and workplace colleagues. CONCLUSIONS: These findings demonstrate that local transmission of MDR-TB is a serious problem in Shenzhen. While most transmission occurred between people who lived distant from each other, there was clear evidence that transmission occurred in schools and workplaces, which should be included as targeted sites for active case finding.The average residential distance between genomic-clustered cases was more than 8 kilometers, while schools and workplaces, identified as sites of transmission in this study, deserve increased vigilance for targeted case finding of multidrug-resistant tuberculosis.

A Highly Reversible Long-Life Li-CO2 Battery with a RuP2 -Based Catalytic Cathode.

Rechargeable Li-CO2 batteries have attracted worldwide attention due to the capability of CO2 capture and superhigh energy density. However, they still suffer from poor cycling performance and huge overpotential. Thus, it is essential to explore highly efficient catalysts to improve the electrochemical performance of Li-CO2 batteries. Here, phytic acid (PA)-cross-linked ruthenium complexes and melamine are used as precursors to design and synthesize RuP2 nanoparticles highly dispersed on N, P dual-doped carbon films (RuP2 -NPCFs), and the obtained RuP2 -NPCF is further applied as the catalytic cathode for Li-CO2 batteries. RuP2 nanoparticles that are uniformly deposited on the surface of NPCF show enhanced catalytic activity to decompose Li2 CO3 at low charge overpotential. In addition, the NPCF its with porous structure in RuP2 -NPCF provides superior electrical conductivity, high electrochemical stability, and enough ion/electron and space for the reversible reaction in Li-CO2 batteries. Hence, the RuP2 -NPCF cathode delivers a superior reversible discharge capacity of 11951 mAh g-1 , and achieves excellent cyclability for more than 200 cycles with low overpotentials (<1.3 V) at the fixed capacity of 1000 mAh g-1 . This work paves a new way to design more effective catalysts for Li-CO2 batteries.

Expression of the Histidine Kinase Gene Sshk Correlates with Dimethachlone Resistance in Sclerotinia sclerotiorum.

Histidine kinases (HK) are implicated in virulence, vegetative mycelial growth, and osmotic and oxidative responses in pathogenic fungi. Our previous work showed that transcriptional levels of the group III HK gene Sshk are higher in field dimethachlone-resistant isolates of Sclerotinia sclerotiorum compared with sensitive isolates. However, it is not clear whether the overexpression of Sshk is the major mechanism for resistance to dimethachlone. In this study, we constructed Sshk silencing and overexpression vectors and assessed dimethachlone resistance levels, virulence, mycelial growth, and sensitivity to osmotic stress for the Sshk-silenced and -overexpression transformants. Overexpression of Sshk resulted in resistance to dimethachlone and increased sensitivity to various stresses and to the cell-wall-perturbing agents sodium dodecyl sulfate (SDS) and Congo red (CR). Compared with the parent isolate, Sshk-silenced transformants had reduced resistance to dimethachlone, significantly higher (P < 0.05) mycelial growth and virulence, and lower sclerotium production, and were less sensitive to various exogenous stresses such as sodium chloride. Compared with the parent sensitive isolate HLJMG1, dimethachlone resistance ratios of the three overexpression transformants ∆C101, ∆C21, and ∆C10 increased 168.1-, 189.5-, and 221.2-fold, respectively. The three overexpression transformants were more sensitive to CR and SDS than their parent isolate. These findings suggest that overexpression of Sshk is a major mechanism for dimethachlone resistance in some isolates of S. sclerotiorum, and that Sshk plays an important role in maintaining the integrity of the cell wall. Our findings reveal a novel molecular mechanism for dimethachlone resistance in plant-pathogenic fungi.

Effects of SHAM on the Sensitivity of Sclerotinia sclerotiorum and Botrytis cinerea to QoI Fungicides.

It is a common practice to add salicylhydroxamic acid (SHAM) into artificial medium in the in vitro sensitivity assay of fungal phytopathogens to the quinone outside inhibitor (QoI) fungicides. The rationale for adding SHAM is to inhibit fungal alternative oxidase, which is presumed to be inhibited by secondary metabolites of plants. Therefore, the ideal characteristics of SHAM should be almost nontoxic to phytopathogens and have no significant effect on control efficacy of fungicides. However, this study showed that the average effective concentration for 50% inhibition (EC50) of mycelial growth values of SHAM were 97.5 and 401.4 µg/ml for Sclerotinia sclerotiorum and Botrytis cinerea, respectively. EC50 values of the three QoI fungicides azoxystrobin, kresoxim-methyl, and trifloxystrobin in the presence of SHAM at 20 and 80 µg/ml for S. sclerotiorum and B. cinerea, respectively, declined by 52.7 to 78.1% compared with those without SHAM. For the dicarboximide fungicide dimethachlone, the average EC50 values in the presence of SHAM declined by 18.2% (P = 0.008) for S. sclerotiorum and 35.9% (P = 0.012) for B. cinerea. Pot experiments showed that SHAM increased control efficacy of the three QoI fungicides against the two pathogens by 43 to 83%. For dimethachlone, SHAM increased control efficacy by 134% for S. sclerotiorum and 86% for B. cinerea. Biochemical studies showed that SHAM significantly inhibited peroxidase activity (P = 0.024) of B. cinerea and esterase activity (P = 0.015) of S. sclerotiorum. The strong inhibitions of SHAM per se on mycelial growth of B. cinerea and S. sclerotiorum and significant influences on the sensitivity of the two pathogens to both the QoI fungicides and dimethachlone as well as inhibitions on peroxidase and esterase indicate that SHAM should not be added in the in vitro assay of sensitivity to the QoI fungicides.

Molecular and biochemical characterization of dimethachlone resistant isolates of Sclerotinia sclerotiorum.

Sclerotinia sclerotiorum is a necrotrophic fungal plant pathogen with a broad host range. The dicarboximide fungicide dimethachlone has been used to control this pathogen for more than a decade and resistance to dimethachlone has recently been reported in China. Compared with sensitive isolates, the three dimethachlone resistant isolates with resistance ratios of 78.3, 85.5, and 94.8 exhibited significantly (P<0.05) higher cell membrane permeability and peroxidase and polyphenol oxidase activities. Dimethachlone at 0.25µg/mL significantly increased cell membrane permeability and enhanced activity of the two enzymes in both resistant and sensitive isolates. There were no significant differences in glycerol or oxalate content between the resistant and sensitive isolates. Dimethachlone treatment increased glycerol content in the resistant isolates and reduced in the sensitive isolates (P<0.01). Sequencing of three genes involved in two-component signal pathway and of three genes in mitogen-activated protein (MAP) kinase cascade demonstrated that the dimethachlone resistant isolates HLJ4 and HLJ6 harbored point mutations of I232T and G1087D, respectively, in the deduce amino acid sequence of the histidine kinase (HK) gene Sshk. HLJ4 had a point mutation of P96L in the deduced amino acid sequence of the MAP kinase-kinase gene SsPbs. The expression levels of the Sshk gene were higher in HLJ4 and HLJ6 than in HLJ3 and the sensitive isolate HLJMG2, and transcription of the Sshk gene was up-regulated by dimethachlone for the three resistant isolates.

Regulation of DNA phosphorothioate modifications by the transcriptional regulator DptB in Salmonella.

DNA phosphorothioate (PT) modifications, with one non-bridging phosphate oxygen replaced with sulfur, are widely but sporadically distributed in prokaryotic genomes. Short consensus sequences surround the modified linkage in each strain, although each site is only partially modified. The mechanism that maintains this low-frequency modification status is still unknown. In Salmonella enterica serovar Cerro 87, PT modification is mediated by a four-gene cluster called dptBCDE. Here, we found that deletion of dptB led to a significant increase in intracellular PT modification level. In this deletion, transcription of downstream genes was elevated during rapid cell growth. Restoration of dptB on a plasmid restored wild-type levels of expression of downstream genes and PT modification. In vitro, DptB directly protected two separate sequences within the dpt promoter region from DNase I cleavage. Each protected sequence contained a direct repeat (DR). Mutagenesis assays of the DRs demonstrated that each DR was essential for DptB binding. The observation of two shifted species by gel-shift analysis suggests dimer conformation of DptB protein. These DRs are conserved among the promoter regions of dptB homologs, suggesting that this regulatory mechanism is widespread. These findings demonstrate that PT modification is regulated at least in part at the transcriptional level.