Identification and Characterization of Pectate Lyase as a Novel Allergen in Artemisia sieversiana Pollen.

INTRODUCTION: Artemisia species are widely spread in north hemisphere. Artemisia sieversiana pollen is one of the common pollen allergens in the north of China. At present, seven allergens were identified and had been listed officially from A. sieversiana pollen, but the remaining allergens are still insufficiently studied, which need to be found. METHODS: Pectate lyase was purified from the extracts of A. sieversiana pollen by anion exchange, size exclusion, and HPLC-hydrophobic interaction chromatography. The gene of A. sieversiana pectate lyase (Art si pectate lyase) was cloned and expressed in Escherichia coli. The enzyme activity and circular dichroism (CD) spectrum of natural and recombinant proteins were analyzed. The allergenicity of Art si pectate lyase was characterized by enzyme-linked immunosorbent assay (ELISA), Western blot, inhibition ELISA, and basophil activation test. The allergen's physicochemical properties, three-dimensional structure, sequence profiles with homologous allergens and phylogenetic tree were analyzed by in silico methods. RESULTS: Natural Art si pectate lyase (nArt si pectate lyase) was purified from A. sieversiana pollen extracts by three chromatographic strategies. The cDNA sequence of Art si pectate lyase had a 1191-bp open reading frame encoding 396 amino acids. Both natural and recombinant pectate lyase (rArt si pectate lyase) exhibited similar CD spectrum, and nArt si pectate lyase had higher enzymatic activity. Moreover, the specific immunoglobulin E (IgE) binding rate against nArt si pectate lyase and rArt si pectate lyase was determined as 40% (6/15) in patients' serum with Artemisia species pollen allergy by ELISA. The nArt si pectate lyase and rArt si pectate lyase could inhibit 76.11% and 47.26% of IgE binding activities to the pollen extracts, respectively. Art si pectate lyase was also confirmed to activate patients' basophils. Its structure contains a predominant motif of classic parallel helical core, consisting of three parallel ß-sheets, and two highly conserved features (vWiDH, RxPxxR) which may contribute to pectate lyase activity. Moreover, Art si pectate lyase shared the highest sequence identity of 73.0% with Art v 6 among currently recognized pectate lyase allergen, both were clustered into the same branch in the phylogenetic tree. CONCLUSION: In this study, pectate lyase was identified and comprehensively characterized as a novel allergen in A. sieversiana pollen. The findings enriched the allergen information for this pollen and promoted the development of component-resolved diagnosis and molecular therapy of A. sieversiana pollen allergy.

Biological control of Magnaporthe oryzae using natively isolated Bacillus subtilis G5 from Oryza officinalis roots.

Rice blast, caused by Magnaporthe oryzae, is a major threat to global rice production causing significant crop losses and impacting grain quality. The annual loss of rice production due to this disease ranges from 10% to 30%. The use of biologically controlled strains, instead of chemical pesticides, to control plant diseases has become a research hotspot. In this study, an antagonistic endophytic bacterial strain was isolated from the roots of Oryza officinalis using the traditional isolation and culture methods. A phylogenetic tree based on 16S RNA and whole-genome sequencing identified isolate G5 as a strain of Bacillus subtilis. This isolate displayed strong antagonistic effects against different physiological strains of M. oryzae. After co-culture in LB medium for 7 days, the inhibition rates of the mycelial growth of four strains of M. oryzae, ZB15, WH97, Guy11, and T-39800E were 98.07 ± 0.0034%, 98.59 ± 0.0051%, 99.16 ± 0.0012%, and 98.69 ± 0.0065%, respectively. Isolate G5 significantly inhibited the formation of conidia of M. oryzae, with an inhibition rate of 97% at an OD600 of 2. Isolate G5 was able to provide 66.81% protection against rice blast under potted conditions. Whole-genome sequencing revealed that the genome size of isolate G5 was 4,065,878 bp, including 4,182 coding genes. Using the anti-SMASH software, 14 secondary metabolite synthesis gene clusters were predicted to encode antifungal substances, such as fengycin, surfactin, and bacilysin. The G5 isolate also contained genes related to plant growth promotion. These findings provide a theoretical basis for expounding the biocontrol mechanisms of this strain and suggest further development of biogenic agents that could effectively inhibit rice blast pathogen growth and reduce crop damage, while being environmentally friendly, conducive to ecological development, and a sustainable alternative to chemical pesticides. This study also enriches the relevant research on endophytes of wild rice, which proves that wild rice is a valuable microbial resource bank.

Degradation of trimethoprim in aqueous by persulfate activated with nanosecond pulsed gas-liquid discharge plasma.

The persulfate activation by nanosecond pulsed gas-liquid discharge (NPG-LD) is employed to degrade the trimethoprim (TMP) in water. The results show that persulfate addition enhances the degradation of TMP by NPG-LD through an obvious synergetic effect. With treatment time of 50 min, the high removal efficiency and energy yield reach 94.6% and 0.57 gkWh-1 in air NPG-LD with the addition of persulfate, respectively, which is 13.5% and 0.09 gkWh-1 higher than that in solo air NPG-LD, respectively. Correspondingly, the calculated synergetic factor achieves 1.62, indicating the synergetic effect is established. The activation mechanism of persulfate by NPG-LD is analyzed by the measurement of reactive species and the effects of radical scavenger addition on TMP removal. It is found that the synergetic effect between NPG-LD and persulfate is attributed to the increased production of OH, H2O2, and . Besides, the TMP degradation by NPG-LD and persulfate synergetic system is influenced by discharge working gas, pulse voltage, addition dosage of persulfate, initial TMP concentration, and initial pH value. Subsequently, the degradation pathway of TMP is analyzed using LC-MS/MS.

Ultra-high synergetic intensity for humic acid removal by coupling bubble discharge with activated carbon.

Humic acid (HA) removal research focuses on the global water treatment industry. In this work, efficient HA degradation with an ultra-high synergetic intensity is achieved by combined bubble discharge with activated carbon (AC). Adding AC to the discharge greatly improves HA removal efficiency and degradation speed; the synergetic intensity reaches 651.52% in the combined system, and the adsorption residual on AC is 4.52%. After 90 min of treatment, the HA removal efficiency reaches 98.90%, 31.29%, and 7.61% in the plasma-AC combined, solo bubble discharge, and solo AC adsorption systems, respectively. During the plasma process, the number of pore structures and active sites and the amount of oxygen-containing functional groups on the AC surface increase, resulting in a higher adsorption capacity to reactive species (H2O2 and O3) and HA and promoting interactions on the AC surface. For HA mineralization, the presence of AC greatly promotes the destruction of aromatic structures and chromophoric HA functional groups.

Nanosecond pulsed uniform dielectric barrier discharge in atmospheric air: A brief spectroscopic analysis.

The paper proposes a simple and convenient approach to represent the discharge uniformity of nanosecond-pulse dielectric barrier discharge (DBD) in air by observation of the ratio of N2+ (B3Σu+ → X3Σg+, 0-0, 391.4 nm) to N2 (C3Πu → B3Πg, 2-5, 394.3 nm) intensities. The DBDs at different pulse peak voltages, discharge gap distances, dielectric materials and thicknesses were investigated by recording their single-pulse-shot discharge images and N2+/N2 ratios to verify the feasibility of the above innovative approach. The results show that the ratios of N2+/N2 are in the range of 0.18-0.6within our experimental parameters, which is respect to the reduced electric field (E/N, where E is the field strength and N is gas number density) strength of 260-440 Td (1 Td = 10-17 V·cm2). And it is indicated that a lower N2+/N2 ratio would be found in a higher pulse peak voltage or/and a lower discharge gap distance, which benefits for improving the discharge uniformity of nanosecond-pulse DBD. The thickness and permittivity of dielectric material also affect the E/N strength and discharge uniformity to a certain extent, but the effects are ambiguous due to additional factors of dielectric materials. In addition, the theoretical basis and application scope of this approach were also discussed.

Spectroscopic and electrical characters of SBD plasma excited by bipolar nanosecond pulse in atmospheric air.

In this paper, an atmospheric surface barrier discharge (SBD) generated by annular electrodes in quartz tube is presented through employing bipolar nanosecond pulse voltage in air. The discharge images, waveforms of pulse voltage and discharge current, and optical emission spectra emitted from the discharges are recorded and calculated. A spectra simulation method is developed to separate the overlap of the secondary diffraction spectra which are produced by grating in monochromator, and N2 (B(3)Πg→A(3)Σu(+)) and O (3p(5)P→3s(5)S2(o)) are extracted. The effects of pulse voltage and discharge power on the emission intensities of OH (A(2)Σ(+)→X(2)Пi), N2(+) (B(2)Σu(+)→X(2)Σg(+)), N2 (C(3)Πu→B(3)Πg), N2 (B(3)Πg→A(3)Σu(+)), and O (3p(5)P→3s(5)S2(o)) are investigated. It is found that increasing the pulse peak voltage can lead to an easier formation of N2(+) (B(2)Σu(+)) than that of N2 (C(3)Πu). Additionally, vibrational and rotational temperatures of the plasma are determined by comparing the experimental and simulated spectra of N2(+) (B(2)Σu(+)→X(2)Σg(+)), and the results show that the vibrational and rotational temperatures are 3250±20K and 350±5K under the pulse peak voltage of 28kV, respectively.

Optical and application study of gas-liquid discharge excited by bipolar nanosecond pulse in atmospheric air.

In this study, a bipolar nanosecond pulse with 20ns rising time is employed to generate air gas-liquid diffuse discharge plasma with room gas temperature in quartz tube at atmospheric pressure. The image of the discharge and optical emission spectra of active species in the plasma are recorded. The plasma gas temperature is determined to be approximately 390K by compared the experimental spectra with the simulated spectra, which is slightly higher than the room temperature. The result indicated that the gas temperature rises gradually with pulse peak voltage increasing, while decreases slightly with the electrode gap distance increasing. As an important application, bipolar nanosecond pulse discharge is used to sterilize the common microorganisms (Actinomycetes, Candida albicans and Escherichia coli) existing in drinking water, which performs high sterilization efficiency.

The influencing factors of nanosecond pulse homogeneous dielectric barrier discharge in air.

In this paper, a bipolar nanosecond high pulse voltage with 20 ns rising time was employed to generate homogeneous dielectric barrier discharges using the plate-plate electrode configuration in air at atmospheric pressure. The effects of pulse peak voltage, gas discharge gap, and dielectric plates made by different materials or thicknesses on the discharge homogeneity, voltage-current waveform, and optical emission spectra were investigated. Results show that aforementioned parameters have a strongly impact on the discharge homogeneity and the optical emission spectra, but it is hard to identify definitely their influences on the discharge voltage-current waveform. Homogeneous discharges were easily observed when using low permittivity dielectric plate and the emission intensity of N2 (C(3)Πu→B(3)Πg, 0-0, 337.1 nm) increases with the rising of pulse peak voltage and the permittivity of dielectric material but decreases with the increasing of gas discharge gap and the dielectric plate thickness. The rotational and vibrational temperatures (Trot and Tvib) were determined at Trot=350±5 K and Tvib=3045 K via fitting the simulative spectra of N2 (C(3)Πu→B(3)Πg, 0-2) with the measured one.

An uniform DBD plasma excited by bipolar nanosecond pulse using wire-cylinder electrode configuration in atmospheric air.

In this study, a bipolar nanosecond pulsed power supply with 15 ns rising time is employed to generate an uniform dielectric barrier discharge using the wire-cylinder electrode configuration in atmospheric air. The images, waveforms of pulse voltage and discharge current, and the optical emission spectra of the discharges are recorded. The rotational and vibrational temperatures of plasma are determined by comparing the simulated spectra with the experimental spectra. The effects of pulse peak voltage, pulse repetition rate and quartz tube diameter on the emission intensities of N2 (C(3)Πu→B(3)Πg, 0-0) and N2(+)B(2)Σu(+)→X(2)Σg(+),0-0 and the rotational and vibrational temperatures have been investigated. It is found that the uniform plasma with low gas temperature can be obtained, and the emission intensities of N2 (C(3)Πu→B(3)Πg, 0-0) and N2(+)B(2)Σu(+)→X(2)Σg(+),0-0 rise with increasing the pulse peak voltage and pulse repetition rate, while decrease as the increase of quartz tube diameter. In addition, under the condition of 28 kV pulse peak voltage, 150 Hz pulse repetition rate and 7 mm quartz tube diameter, the plasma gas temperature is determined to be 330 K. The results also indicate that the plasma gas temperature keep almost constant when increasing the pulse peak voltage and pulse repetition rate but increase with the increase of the quartz tube diameter.

A large-area diffuse air discharge plasma excited by nanosecond pulse under a double hexagon needle-array electrode.

A large-area diffuse air discharge plasma excited by bipolar nanosecond pulse is generated under a double hexagon needle-array electrode at atmospheric pressure. The images of the diffuse discharge, electric characteristics, and the optical emission spectra emitted from the diffuse air discharge plasma are obtained. Based on the waveforms of pulse voltage and current, the power consumption, and the power density of the diffuse air discharge plasma are investigated under different pulse peak voltages. The electron density and the electron temperature of the diffuse plasma are estimated to be approximately 1.42×10(11) cm(-3) and 4.4 eV, respectively. The optical emission spectra are arranged to determine the rotational and vibrational temperatures by comparing experimental with simulated spectra. Meanwhile, the rotational and vibrational temperatures of the diffuse discharge plasma are also discussed under different pulse peak voltages and pulse repetition rates, respectively. In addition, the diffuse air discharge plasma can form an area of about 70×50 mm(2) on the surface of dielectric layer and can be scaled up to the required size.

[Optical diagnosis of large area homogenous dielectric barrier discharge in nitrogen at atmospheric pressure].

In the present study, dielectric barrier homogenous discharge in nitrogen was obtained between large plate electrodes (150 x 300 mm) at atmospheric pressure and the emission spectra of N2 (C3pi(u) --> B3 pi(g)) and N2+ (B2 sigma(u)+ --> X2 sigma(g)+ 0-0 391.4 nm) were recorded. It was found that both the emission intensities of N2 (C3 pi(u) --> B3 pi(g) and N2+ (B2 sigma(u)+ --> X2 sigma(g)+ 0-0 391.4 nm) increase with the rising of the applied voltage and the driving frequency, respectively. The main physicochemical formation mechanism of N2+ (B2 sigma(u)+) in N2 and He+N2 mixtures homogenous discharge was discussed, and the penning ionization was proved to be the dominant formation mechanism.