Ultrafast protein digestion using an immobilized enzyme reactor following high-resolution mass spectrometry analysis for rapid identification of abrin toxin.

Abrin toxin, highly dangerous with an estimated human lethal dose of 0.1-1 µg per kg body weight, has attracted much attention regarding criminal and terroristic misuse over the past decade. Therefore, developing a rapid detection method for abrin toxin is of great significance in the field of biosecurity. In this study, based on the specific dissociation method of an immobilized enzyme reactor, the trypsin immobilized reactor Fe3O4@CTS-GA-Try was prepared to replace free trypsin, and the immobilized enzyme digestion process was systematically investigated and optimized by using bovine serum albumin as the simulant of abrin. After 5 min one-step denaturation and reduction, a satisfactory peptide number and coverage were yielded with only 15 s assisted by an ultrasound probe to identify model proteins. Subsequently, abrin was rapidly digested using the established method, resulting in a stable and highly reproducible characteristic peptide number of 39, which can be analyzed by nanoelectrospray ionization coupled with high-resolution mass spectrometry. With the acquisition mode of full MS scan coupled with PRM, not only MS spectroscopy of total abrin peptides but also the corresponding MS/MS spectroscopy of specific abrin peptides can achieve the characteristic detection of abrin toxin and its different isoforms in less than 10 minutes, with high repeatability. This assay provides a universal platform and has great potential for the development of on-site detection and rapid mass spectrometric analysis techniques for macromolecular protein toxins and can further be applied to the integrated detection of chemical and biological agents.

Accurate Detection of G-Series Agent Simulants by Pseudo-Multiple Reaction Monitoring on Miniature LIT-MS.

Rapid and accurate on-site detection of chemical warfare agents (CWAs) could defend military and civilian populations against current and emerging chemical weapons. With the development of ambient ionization and linear ion trap technology, the rapid and accurate quantitative determination method of CWAs based on direct ionization and multistage mass spectrometry has attracted widespread attention. In this study, a microliter electrospray ionization-miniature linear ion trap mass spectrometry (LIT-MS) instrument was designed and constructed, and the effects of quadrupole enhanced dipole resonance excitation on the resolution and sensitivity were investigated; consequently, the parameters of CWAs detection were optimized. Based on the broad time-frequency ion excitation technology, accurate multiple reaction monitoring (MRM) quantitative analysis of DMMP (G-series agent simulants, m/z 125 → m/z 93) was obtained. The linear correlation coefficient in the concentration range of 1 to 20 µg/mL could reach 99.02%, and the relative standard deviations (RSD) of continuous repeatability, interday repeatability, and intraday repeatability were all less than 10%. The results showed that the accurate pseudo-MRM detection method based on miniature linear ion trap mass spectrometry for CWAs detection was feasible.

Temperature-Tuning Desorption Ionization for Ambient Mass Spectrometry.

In the past decade, mass spectrometry (MS) has been widely used for a broad range of on-site applications. This is largely attributed to the rapid advancement of technologies, such as ambient ionization and mass spectrometer miniaturization. Here, we report the development of the temperature-tuning desorption ionization (TTDI) method for versatile on-site applications using a miniature MS system. A unique feature of TTDI is its dynamic temperature range applicable from 30 to 800 °C, which enables optimal desorption ionization applied for chemical and biological compounds through tuning the temperature at the sampling spot. The versatility of TTDI has been demonstrated through on-site MS analysis of a variety of samples, such as explosives on surfaces, drugs of abuse in biofluids, and screening biomarkers in tissues.

Experimental Study on Bubble Dynamics and Mass Transfer Characteristics of Coaxial Bubbles in Petroleum-Based Liquids.

Petroleum-based liquids are from an important petroleum-based polymer, whose application and preparation involve multiple operations related to gas-liquid two-phase flow. Due to insufficient research on gas-liquid two-phase flow, there is a gap in bubble dynamics and mass transfer characteristics in petroleum-based liquids. Accordingly, we have systematically investigated the bubble formation process, bubble rising dynamics, and mass transfer of coaxial bubbles. Herein, the contour of bubbles was obtained for analyzing the bubble formation process. It was found that the increase of gas flow rate contributed to the increase of bubble generation size, while the liquid viscosity had an inhibitory influence on the increase of bubble generation size. Moreover, the variation of bubble rising velocity was considered and the force analysis of the rising bubble was provided. A new model of drag coefficient applicable to petroleum-based liquids was proposed. Finally, variations in the amount of dissolved oxygen in the liquid were measured to analyze the mass transfer characteristics. The increase in nozzle inner diameter and gas flow rate both promoted mass transfer, but the increased liquid viscosity hindered mass transfer.

Ultracompact single-layer optical MEMS accelerometer based on evanescent coupling through silicon nanowaveguides.

In this paper, a novel optical MEMS accelerometer is proposed based on evanescent coupling between parallel silicon nanowaveguides. The coupling length between nanowaveguides changes due to the input acceleration, leading to a great change of coupling efficiency. As a result, the applied acceleration can be obtained by measuring the transmission of waveguiding light. Simulation results with optical displacement sensing sensitivity of 32.83%/[Formula: see text]m within measurement range of 1.68 g is obtained. This design shows high compactness with no need of assembly, suggesting great potential in applications such as integrated photonic circuits.

Investigation of a Complex Reaction Pathway Network of Isobutane/2-Butene Alkylation by CGC-FID and CGC-MS-DS.

The mechanism of reaction in isobutane/2-butene alkylation systems is extremely complicated, accompanied by numerous side reactions. Therefore, a comprehensive understanding of the reaction pathways in this system is essential for an in-depth discussion of the reaction mechanism and for improving the selectivity of the major products (clean fuel blend components). The alkylation of isobutane/2-butene was studied using a self-made intermittent reaction device with a metering, cooling, reaction, vacuum and analysis system. The alkylates were qualitatively and quantitatively analyzed using a capillary gas chromatography-mass spectrometry-data system (CGC-MS-DS) and capillary gas chromatography with flame ionization detection (CCGC-FID), respectively, and the precision and recovery of the quantitative analytical methods were verified. The results showed that the relative standard deviation (RSD) of the standard sample was below 0.78%, and the recoveries were from 98.53% to 102.85%. Under the specified reaction conditions, 79 volatile substances were identified from the alkylates, and the selectivity of C8 and trimethylpentanes (TMPs) reached 63.63% and 53.81%, respectively. The changes of the main chemical components in the alkylation reaction with time were tracked and analyzed, based on which reaction pathways were determined, and a complex reaction network containing the main products' and the by-products' generation pathway was constructed.

Survey on antibiotic regimens for necrotizing enterocolitis prescribed by Chinese pediatricians in 2020.

Background: Necrotizing enterocolitis (NEC) is a serious intestinal inflammatory disease in neonates, and intravenous antibiotics constitute the main therapeutic strategy. Studies have shown that substantial variation in the selection of antibiotic regimens for NEC remains in many countries. The variability in antibiotic therapy selection in China is unclear. Methods: A cross-sectional study using an online questionnaire regarding antibiotic regimens for NEC was conducted among pediatricians working in tertiary hospitals in China. Results: A total of 284 pediatricians from 29 provinces completed the survey; 37.9% of them administered one antibiotic, 56.7% administered two antibiotics and 2.4% administered three antibiotics. The top three single-antibiotic regimens for NEC were beta-lactamase inhibitors (n=66, 41.5%), carbapenems (n=46, 28.9%) and cephalosporins (n=33, 20.8%). Twenty combinations of two antibiotics were identified, and the top three combinations were beta-lactamase inhibitors and antianaerobic agents (n=49, 19.6%), carbapenems and glycopeptides (n=42, 16.8%), and cephalosporins and antianaerobic agents (n=37, 14.8%). Regarding the therapeutic duration of antibiotic treatment, 77.5% (n=220) of the pediatricians chose 5-10 days for stage II NEC, and 79.6% (n=226) chose 7-14 days for stage III NEC. Forty-three percent (n=122) of the respondents preferred to use carbapenems when NEC was diagnosed, and 83.3% (n=135) adjusted the antibiotics to carbapenems if a patient's medical condition deteriorated or if the therapeutic efficacy was poor. Conclusion: This survey revealed that carbapenems were the most common antibiotics selected when NEC was diagnosed or a patient's condition deteriorated, but the duration of the course of treatment for NEC varied substantially among doctors.

The ionization process of chemical warfare agent simulants in low temperature plasma ionization.

The application of low-temperature plasma ionization technology in the chemical warfare agent detection was mostly focused on the research of rapid detection methods. Limited studies are available on the ionization process of chemical warfare agents in low temperature plasma. Through the intensity of protonated molecules of dimethyl methylphosphonate (DMMP) in different solvents including methanol, deuterated methanol (methanol-D4), pure water, and deuterium oxide (water-D2), it was concluded that the water molecule in the air provides the hydrogen ion (H+) needed for ionization. The product ion spectra and the collision-induced dissociation processes of protonated molecules of nerve agent simulants, including DMMP, diethyl methanephosphonate (DEMP), trimethyl phosphate (TMP), triethyl phosphate (TEP), tripropyl phosphate (TPP), and tributyl phosphate (TBP) were analyzed. Results revealed that H+ mostly combined with phosphorus oxygen double bond (P = O) in the low-temperature plasma ionization. By analyzing the peak intensity distribution of product ions of protonated molecules, the presence of proton and charge migration in the low temperature plasma ionization and collision-induced dissociation were researched. This study could provide technical guidance for the rapid and accurate detection of chemical warfare agents through low temperature plasma ionization-mass spectrometry.

Early type I IFN blockade improves the efficacy of viral vaccines.

Type I interferons (IFN-I) are a major antiviral defense and are critical for the activation of the adaptive immune system. However, early viral clearance by IFN-I could limit antigen availability, which could in turn impinge upon the priming of the adaptive immune system. In this study, we hypothesized that transient IFN-I blockade could increase antigen presentation after acute viral infection. To test this hypothesis, we infected mice with viruses coadministered with a single dose of IFN-I receptor-blocking antibody to induce a short-term blockade of the IFN-I pathway. This resulted in a transient "spike" in antigen levels, followed by rapid antigen clearance. Interestingly, short-term IFN-I blockade after coronavirus, flavivirus, rhabdovirus, or arenavirus infection induced a long-lasting enhancement of immunological memory that conferred improved protection upon subsequent reinfections. Short-term IFN-I blockade also improved the efficacy of viral vaccines. These findings demonstrate a novel mechanism by which IFN-I regulate immunological memory and provide insights for rational vaccine design.

Complete genome sequence of the lytic cold-active Pseudomonas fluorescens bacteriophage VSW-3 from Napahai plateau wetland.

The lytic cold-active bacteriophage VSW-3, belonging to the Podoviridae family and infecting the host Pseudomonas fluorescens SW-3, was isolated from the Napahai plateau wetland in China. With the development of sequencing technology, the study of Pseudomonas genomic diversity has increased; however, knowledge of cold-active phages infecting Pseudomonas is limited. The newly sequenced phage VSW-3 was classified based on virion morphology by transmission electron microscope. Sequence analysis revealed that the genome size was 40,556 bp with an overall GC content of 57.54 % and 46 open reading frames. The genome was organized into several modules containing genes for packaging, structural proteins, replication/transcription, and phage lysis. The sequence contained 45 potential promoters, 3 transcription terminators, and yet no tRNAs. This is the first report of cold-active Pseudomonas fluorescens bacteriophage genome sequencing.

A novel asymmetrical arc-shaped electrode ion trap for improving the performance of a miniature mass spectrometer.

RATIONALE: The pivotal challenge associated with miniature mass analyzers is their proper design and construction without sacrificing performance. In order to analyze and improve the performance of a miniature linear ion trap with odd and even multipole fields, we designed a novel asymmetrical arc-shaped electrode ion trap (AAEIT), and tested the properties of AAEITs with different dimensions. METHODS: A series of asymmetrical ion traps using arc-shaped electrodes were designed to optimize the properties (resolutions and intensity) of the coupling effects between odd and even multipole fields. Using arginine and reserpine, we evaluated the performance of mass resolution, ion intensity ratio and deduced the collision-induced dissociation (CID) efficiency using a self-constructed electrospray ionization mass spectrometry (ESI-MS) platform. RESULTS: An AAEIT with field radius dimensions of 5 mm × 5.75 mm exhibits a good performance: its maximum resolution of 833 (FWHM) at m/z 175 was achieved for the side of small electrode. With this AAEIT, a tandem mass (MS/MS) capability with 91.0% CID efficiency was obtained with reserpine (m/z 609). CONCLUSIONS: The results indicated that the AAEIT, comprising both odd and even multipole fields, could act as a qualified linear ion trap mass analyzer with compact structure, high resolution, and high tandem mass analysis efficiency. It has a great potential in miniature mass spectrometry.

Determining the age at death of females in the Chinese Han population: using quantitative variables and statistical analysis from pubic bones.

Determining the age at death of females by skeletal features is extremely difficult and important, both in forensics and in physical anthropology. Our previous study of male pubic symphysis suggests that the indicators of morphological changes provide the best results for personal age identification. The indicators that help deduce of the age of females was approximately the same as males except for two specific indicators, which are bone density of the symphysial surface and viz. Viz are ridges and furrows on the symphysial surface, ossific nodules, lower extremities, ventral and ridge of the rampart, dorsal margin, ventral beveling and general macroscopic changes on the symphysial surface. Samples were drawn from 338 female individuals. The study procedures are as follows: Firstly, we examined the morphological features of pubic symphysis using the criteria similar to Hanihara's, Gilbert-McKern and Suchey-Brooks's methods. Secondly, we evaluated each stage with an appropriate score. Thirdly, we deduced four equations to assess the morphological features of the ages of females by statistic analysis. The results were discussed by comparing with Hanihara's, Gilbert-McKern and Suchey-Brooks's methods. The results were consistent and the developing methods for determining the age of death of females were produced.

Determination of male age at death in Chinese Han population: using quantitative variables statistical analysis from pubic bones.

Determination of age at death by skeletal features is difficult, but important, both in forensic and physical anthropology. Pubic symphysis is a widely accepted morphological indicator, which provides the best result for personal age identification. Age prediction in this study used nine indicators of morphological changes, viz: ridges and furrows on the symphysial surface, ridge of pubic tubercle, lower extremity, ventral beveling, ossific nodules, dorsal margin, ventral rampart, general macroscopic changes of symphysial surface, and bone density of the symphysial surface. Samples were drawn from 262 male individuals. The study procedures are as follows: Firstly, we examined the morphological features of pubic symphysis using criteria similar to Hanihara's and Suchey-Brooks (SB). Secondly, we evaluated each stage with an appropriate score. Thirdly, we deduced four equations of male age assessment of morphological features by statistic analysis. The results were compared with Hanihara's and Suchey-Brooks method; the similarities and differences among them were discussed objectively. The method produced consistent results as well as developing the criteria of Hanihara's and Suchey-Brooks'.