Molecular characterization, expression and antibacterial function of a macin, HdMac, from Haliotis discus hannai.

Macins are a family of antimicrobial peptides, which play multiple roles in the elimination of invading pathogens. In the present study, a macin was cloned and characterized from Pacific abalone Haliotis discus hannai (Designated as HdMac). Analysis of the conserved domain suggested that HdMac was a new member of the macin family. In non-stimulated abalones, HdMac transcripts were constitutively expressed in all five tested tissues, especially in hemocytes. After Vibrio harveyi stimulation, the expression of HdMac mRNA in hemocytes was significantly up-regulated at 12 hr (P < 0.01). RNAi-mediated knockdown of HdMac transcripts affected the survival rates of abalone against V. harveyi. Moreover, recombinant protein of HdMac (rHdMac) exhibited high antibacterial activities against invading bacteria, especially for Vibrio anguillarum. In addition, rHdMac possessed binding activities towards glucan, lipopolysaccharides (LPS), and peptidoglycan (PGN), but not chitin in vitro. Membrane integrity analysis revealed that rHdMac could increase the membrane permeability of bacteria. Meanwhile, both the phagocytosis and chemotaxis ability of hemocytes could be significantly enhanced by rHdMac. Overall, the results showed that HdMac could function as a versatile molecule involved in immune responses of H. discus hannai.

Interaction between Serum Cotinine and Magnesium Intake with Childhood Asthma: A Cross-Sectional Study.

INTRODUCTION: Serum cotinine and magnesium intake are often associated with childhood asthma. This study evaluated the interaction between serum cotinine and magnesium intake and childhood asthma. METHODS: This cross-sectional study included 14,159 subjects from the National Health and Nutrition Examination Survey 2005-2018. Serum cotinine levels were classified according to the lower quartile: ≤0.2089 ng/mL as low level and >0.2089 ng/mL as high level. Magnesium intake was categorized as high (>98 mg/1,000 kcal) or low level (≤98 mg/1,000 kcal) based on the upper quartile. Weighted logistic regression analyses were adopted to analyze the association between cotinine, magnesium intake, and childhood asthma. Additionally, the combined effect of cotinine and magnesium intake on childhood asthma risk was examined. The stratified analyses were based on gender, body mass index, and family history of asthma to further examine the relationship between cotinine, magnesium intake, and childhood asthma. RESULTS: The prevalence of asthma was approximately 17.56%. Compared to low-level cotinine, high-level cotinine was associated with asthma (odds ratio [OR] = 1.25, 95% confidence interval (CI): 1.04-1.50). Low-level magnesium intake was related to asthma compared with high-level magnesium intake (OR = 1.21, 95% CI: 1.04-1.40). Using interaction analysis, we also found that the combined effect of cotinine and magnesium intake was associated with childhood asthma risk, and the interaction between high-level cotinine and low-level magnesium intake was associated with the highest risk of childhood asthma (OR = 1.35, 95% CI: 1.04-1.74). Additionally, this interaction was also found in males, overweight/non-overweight, and those with family history of asthma. CONCLUSION: There was an interaction between serum cotinine and magnesium intake on childhood asthma. The results suggested that implementing smoking bans in certain settings (e.g., communities, schools) and promoting the consumption of magnesium-rich foods may be effective strategies for preventing childhood asthma.

Multitarget-Tracking Method Based on the Fusion of Millimeter-Wave Radar and LiDAR Sensor Information for Autonomous Vehicles.

Multitarget tracking based on multisensor fusion perception is one of the key technologies to realize the intelligent driving of automobiles and has become a research hotspot in the field of intelligent driving. However, most current autonomous-vehicle target-tracking methods based on the fusion of millimeter-wave radar and lidar information struggle to guarantee accuracy and reliability in the measured data, and cannot effectively solve the multitarget-tracking problem in complex scenes. In view of this, based on the distributed multisensor multitarget tracking (DMMT) system, this paper proposes a multitarget-tracking method for autonomous vehicles that comprehensively considers key technologies such as target tracking, sensor registration, track association, and data fusion based on millimeter-wave radar and lidar. First, a single-sensor multitarget-tracking method suitable for millimeter-wave radar and lidar is proposed to form the respective target tracks; second, the Kalman filter temporal registration method and the residual bias estimation spatial registration method are used to realize the temporal and spatial registration of millimeter-wave radar and lidar data; third, use the sequential m-best method based on the new target density to find the track the correlation of different sensors; and finally, the IF heterogeneous sensor fusion algorithm is used to optimally combine the track information provided by millimeter-wave radar and lidar, and finally form a stable and high-precision global track. In order to verify the proposed method, a multitarget-tracking simulation verification in a high-speed scene is carried out. The results show that the multitarget-tracking method proposed in this paper can realize the track tracking of multiple target vehicles in high-speed driving scenarios. Compared with a single-radar tracker, the position, velocity, size, and direction estimation errors of the track fusion tracker are reduced by 85.5%, 64.6%, 75.3%, and 9.5% respectively, and the average value of GOSPA indicators is reduced by 19.8%; more accurate target state information can be obtained than a single-radar tracker.

Sulfuric acid-dimethylamine particle formation enhanced by functional organic acids: an integrated experimental and theoretical study.

Atmospheric new particle formation (NPF), which has been observed globally in clean and polluted environments, is an important source of boundary-layer aerosol particles and cloud condensation nuclei, but the fundamental mechanisms leading to multi-component aerosol formation have not been well understood. Here, we use experiments and quantum chemical calculations to better understand the involvement of carboxylic acids in initial NPF from gas phase mixtures of carboxylic acid, sulfuric acid (SA), dimethylamine, and water. A turbulent flow tube coupled to an ultrafine condensation particle counter with particle size magnifier has been set up to measure NPF. Experimental results show that pyruvic acid (PA), succinic acid (SUA), and malic acid (MA) can enhance sulfuric acid-dimethylamine nucleation in the order PA < SUA < MA with a greater enhancement observed at lower SA concentrations. Computational results indicate that the carboxylic and hydroxyl groups are related to the enhancement. This experiment-theory study shows the formation of multi-component aerosol particles and the role of the organic functional group, which may aid in understanding the role of organics in aerosol nucleation and growth in polluted areas, and help to choose organic molecules of specific structures for simulation.

The induction of inflammation by the cGAS-STING pathway in human dental pulp cells: A laboratory investigation.

AIM: To explore the presence of the cGAS-STING inflammatory pathway in human pulp tissue and human dental pulp cells (HDPCs). METHODOLOGY: Pulp tissue was collected from freshly extracted human healthy third molars or third molars with irreversible pulpitis. Quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunoassay (ELISA) were performed to assess IFN-ß, TNF and IL-6. Human dental pulp cells prepared from healthy human pulp tissues were transfected with interferon stimulatory DNA (ISD), bacterial genomic DNA, bacterial cyclic dinucleotides c-di-AMP, c-di-GMP or host cyclic dinucleotide cGAMP. SiRNA was used to knock down the endogenous cGAS or STING. G140 and H-151 were used to inhibit cGAS and STING respectively. Amlexanox and BAY 11-7082 were used to inhibit TBK1 and NF-κB respectively. qRT-PCR and ELISA were performed to detect the level of IFN-ß, TNF and IL-6. Western blot was performed to evaluate the TBK1, IRF3 and p65 phosphorylation. The Student's t-test and one-way anova were used for statistical analysis. RESULTS: IFN-ß, TNF and IL-6 were up-regulated in the inflamed human dental pulp tissue. CGAS and STING mRNA were increased in the inflamed human dental pulp tissue and detected in HDPCs prepared from healthy human pulp tissues. ISD transfection induced TBK1, IRF3 and p65 phosphorylation as well as IFN-ß, TNF and IL-6 production. IFN-ß, TNF and IL-6 production were also induced by transfection of bacterial and host cyclic dinucleotides or bacteria DNA. ISD or bacteria DNA transfection elevated the intracellular levels of cGAMP. Knock-down of cGAS or STING, as well as using cGAS inhibitor G140 or STING inhibitor H-151 abolished the IFN-ß, TNF and IL-6 production induced by ISD transfection. Knock-down of STING or using STING inhibitor H-151 abolished the IFN-ß, TNF and IL-6 induction by transfection of bacterial and host cyclic dinucleotides. Both Amlexanox and BAY 11-7082 inhibited IFN-ß, TNF and IL-6 production triggered by ISD and cyclic dinucleotides transfection. CONCLUSIONS: Human dental pulp cells expressed an intact cGAS-STING signalling axis. The cGAS-STING signalling axis may play an important role in pulp inflammation and immune defence.

The cysteine protease ATG4B of Trichinella spiralis promotes larval invasion into the intestine of the host.

The cysteine proteases of parasites are vital contributors that induce parasite migration to and invasion of host tissue. In this study, we analysed the cysteine protease ATG4B of Trichinella spiralis (TsATG4B) isolated from the soluble proteins of Trichinella spiralis (T. spiralis) adult worms to ascertain its biochemical properties and functions during invasion into the intestine of the host. The 43 kDa recombinant cysteine protease ATG4B protein (rTsATG4B) consists of a conserved peptidase_C54 domain and was expressed in Escherichia coli. Gelatine zymography showed that rTsATG4B could hydrolyse gelatine and that the hydrolytic activity was prevented by the cysteine protease inhibitor E-64 (pH 5.2). Immunofluorescence assays showed that TsATG4B is expressed at different stages and is localized at the cuticles and stichosomes of worms. Far-Western blotting and confocal microscopy revealed that rTsATG4B interacts with intestinal epithelial cells (IECs) and that it was subcellularly localized to the membrane and cytoplasm in IECs. Real­time quantitative PCR (qPCR) results indicated that the transcription level of the TsATG4B gene was the higher in 6-day-old adult worms (6 days AW) than in any other stage. An in vitro larval invasion assay verified that rTsATG4B promoted larval invasion and that invasion was inhibited when rTsATG4B was pre-incubated with E-64, whereas anti-rTsATG4B serum inhibited larval invasion in a dose-dependent manner. Collectively, these results suggested that the enzymatic activity of TsATG4B significantly influences the hydrolysis process, which is necessary for larval invasion of the host intestinal epithelium.

Hydration of 3-hydroxy-4,4-dimethylglutaric acid with dimethylamine complex and its atmospheric implications.

Atmospheric aerosols have a tremendous influence on visibility, climate, and human health. New particle formation (NPF) is a crucial source of atmospheric aerosols. At present, certain field observations and experiments have discovered the presence of 3-hydroxy-4,4-dimethylglutaric acid (HDMGA), which may participate in NPF events. However, the nucleation mechanism of HDMGA has not been clearly understood. In addition, dimethylamine (DMA) is an important precursor of nucleation. The nucleation mechanism involving HDMGA and DMA has not been studied. In this study, the most stable structures of (HDMGA)(H2O)n (n = 0-3) and (HDMGA)(DMA)(H2O)n (n = 0-3) were obtained by using M06-2X coupled with the 6-311++G(3df,3pd) basis set. The α-carboxyl group is directly attached to the amino group in all the most stable configurations. Proton transfer enhances the strength of a hydrogen bond, as well as promotes the generation of a global minimum structure. Temperature has a considerable influence on the distribution of isomers for (HDMGA)(DMA)(H2O)3 as compared to the other investigated clusters. The Gibbs free energy values reveal that most of the clusters can exist in NPF, except for (HDMGA)(H2O)1. The process of adding a cluster of (H2O)n more likely occurs in the atmosphere than gradually adding a single water molecule.

Synergistic Effect of Ammonia and Methylamine on Nucleation in the Earth's Atmosphere. A Theoretical Study.

Ammonia and amines are important common trace atmospheric species that can enhance new particle formation (NPF) in the Earth's atmosphere. However, the synergistic effect of these two bases involving nucleation is still lacking. We studied the most stable geometric structures and thermodynamics of quaternary (NH3)(CH3NH2)(H2SO4) m(H2O) n ( m = 1-3, n = 0-4) clusters at the PW91PW91/6-311++G(3df,3pd) level of theory for the first time. We find that the proton transfer from H2SO4 molecule to CH3NH2 molecule is easier than to NH3 molecule in the free or hydrated H2SO4-base clusters, and thus leads to the stability. The energetically favorable formation of the (NH3)(CH3NH2)(H2SO4) m(H2O) n ( n = 0-4) clusters, by hydration or attachment of base or substitution of ammonia by methylamine at 298.15 K, indicate that ammonia and methylamine together could enhance the stabilization of small binary clusters. At low RH and an ambient temperature of 298.15 K, the concentration of total hydrated (NH3)(CH3NH2)(H2SO4)2 clusters could reach that of total hydrated (NH3)(H2SO4)2 clusters, which is the most stable ammonia-containing cluster. These results indicate that the synergistic effect of NH3 and CH3NH2 might be important in forming the initial cluster with sulfuric acid and subsequently growth process. In addition, the evaporation rates of (NH3)(CH3NH2)(H2SO4)(H2O), (NH3)(CH3NH2)(H2SO4)2 and (NH3)(CH3NH2)(H2SO4)3 clusters, three relative abundant clusters in (NH3)(CH3NH2)(H2SO4) m(H2O) n system, were calculated. We find the stability increases with the increasing number of H2SO4 molecules.

Knockdown resistance of Anopheles sinensis in Henan province, China.

BACKGROUND: Vivax malaria was historically epidemic in Henan Province of China and Anopheles sinensis was the main vectors and poor farming communities bare the greatest burden of disease. Knockdown resistance in An. sinensis is one of the mechanisms of resistance against pyrethroids. In the present study, the frequency of mutations from An. sinensis was examined in Henan province, China. METHODS: Anopheles was collected from Kaifeng, Tongbai, Tanghe, Pingqiao, Shihe, and Yongcheng counties of Henan province in 2013. Molecular identification of Anopheles species was conducted by polymerase chain reaction (PCR) amplifying the internal transcribed spacer 2 (ITS2). Part of the IIS6 domain of the para-type sodium channel protein gene was polymerase chain reaction-amplified and directly sequenced. Frequency and geographic difference of kdr gene mutant types were analysed. RESULTS: 208 Anopheles were received molecular identification, of which 169 (81.25%) were An. sinensis, 25 (12.02%) were Anopheles yatsushiroensis, and 12 (5.77%) were Anopheles lesteri. A 325 bp fragment of the para-type sodium channel gene including position 1014 was successfully sequenced from 139 Anopheles, of which 125 (89.93%) were An. sinensis, 12 (8.63%) were An. yatsushiroensis, 2 (1.44%) were An. lesteri. The molecular analyses revealed that mutations existed at codon 1014 in An. sinensis but not in An. yatsushiroensis and An. lesteri. Frequency of kdr mutation was 73.60% (92/125) from population of An. sinensis in Henan province, of which L1014F (TTT + TTC) allele frequencies accounted for 46.40% (58/125), and was higher than that of L1014C(TGT) which accounted for 27.20% (34/125) ( χ2 = 55.423, P < 0.001). The frequency of kdr mutation in Kaifeng county was 100% (49/49), and was higher than that of 37.93% (11/29) in Tongbai, 54.55% (6/11) in Pingqiao, 50.00% (3/3) in Shihe, and 62.50% (10/16) in Yongcheng county, respectively (χ2 = 39.538, P < 0.001; χ2 = 24.298, P < 0.001; χ2 = 25.913, P < 0.001; χ2 = 20.244, P < 0.001). While 92.86% (13/14) frequency of kdr mutation was found in Tanghe county, which was higher than that in Tongbai county (χ2 = 11.550, P = 0.0018). CONCLUSIONS: A high frequency of kdr gene mutations from population of An. sinensis in Henan province was found.

A new potential energy surface and microwave and infrared spectra of the He-OCS complex.

A new high quality potential energy surface for the He-OCS van der Waals complex was calculated using the CCSD(T) method and avqz+33221 basis set. It is found that the global minimum energy is -51.33 cm(-1) at R(e) = 6.30a0 and θ(e) = 110.0°, the shallower minimum is located at R = 8.50a0 and θ = 0° with well depth -32.26 cm(-1). Using the fitted potential energy surface, we have calculated bound energy levels of the He-OCS, He-O(13)CS, He-OC(34)S, and (3)He-OCS complexes. The theoretical results are all in better agreement compared to previous theoretical work.