Fast analysis of selected compounds in inhaled and exhaled vapor phase of cigarette smoke to evaluate components retained in the upper respiratory tract.

RATIONALE: The aim of this work is to use a new design of online sampling photoionization mass spectrometer to analyze chemical ingredients in inhaled and exhaled cigarette smoke directly without separation. METHODS: Based on vacuum ultraviolet photoionization time-of-flight mass spectrometry (VUV-PI-TOFMS) and a sampling system, a newly developed rapid online sampling design approach was used for the upper respiratory tract retention study of gaseous mainstream cigarette smoke components during smoking. The cigarette smoke inhaled or exhaled by seven subjects who displayed three different smoking patterns was directly sampled into a vacuum chamber, photoionized, and analyzed using TOFMS. RESULTS: Fourteen species, comprising aldehydes, ketones, phenol, methanethiol, nitrogen-containing heterocyclic compounds and unsaturated hydrocarbons, were identified in the cigarette smoke obtained from Virginia-type cigarettes. The upper respiratory tract results for these compounds were similar for smokers with the three different smoking patterns: aldehyde and ketone constituents had a high retention level of more than 60%; phenol, methanethiol, and nitrogen-containing heterocyclic compounds were retained at between 30% and 70%; and the retention of unsaturated hydrocarbons was about 20%-60%. The retention trend of the same smoke components in Virginia-type cigarettes by subjects from the three smoking patterns (A, B, and C) was consistent, and the retentions all increased with increased smoking age (A < B < C). CONCLUSIONS: This is the first report of a new online sampling design approach to the study of cigarette smoke components in inhaled and exhaled breath, to evaluate components retained in the upper respiratory tract by subjects with different smoking patterns. This method has good repeatability, and the results indicated that this is a very promising tool for the study of the retention of cigarette smoke constituents.

Dissociative photoionization of ß-pinene: an experimental and theoretical study.

We investigated the photoionization and dissociation photoionization of the ß-pinene molecular using time-of-flight mass spectrometry with a tunable vacuum ultraviolet source in the region from 8.00eV to 15.50eV. The experimental ionization energy (IE) value is 8.60eV using electron impact as the ionization source which is not in good agreement with theoretical value (8.41 eV) with a G3MP2 method. We obtained the accurate IE of ß-pinene (8.45 ± 0.03eV) derived from the efficiency spectrum which is in good agreement with the theoretical value (8.38eV) of the CBS-QB3 method. We elucidated the dissociation pathways of primary fragment ions from the ß-pinene cation on the basis of experimental observations in combination with theoretical calculations. Most of the dissociation pathways occur via a rearrangement reaction prior to dissociation. We also determined the structures of the transition states and intermediates for those isomerization processes.

Identification of cigarettes with different grades by using FTIR microspectroscopy.

In this paper, FTIR microspectroscopy was used to identify the cigarettes of three different grades such as Jinwan cigarettes (J group), Yinxiangyipin cigarettes (Y group) and Hongsanhuan cigarettes (H group). IR spectra and peak-area ratios (A2923 /A816, A1601 /A2923, A1601/A920 and A1072/A816) revealed significant differences among H, Y and J groups, reflecting the changes in chemical compositions with increased grade. Discriminant analysis was carried out on basis of the above peak-area ratios, achieving 100% accuracy for identification of H, Y and J groups. Principal component analysis (PCA) showed that carbohydrates and proteins were closely related to the quality of the cigarettes. In addition, curve fitting further confirmed that the structure of carbohydrates underwent changes due to the quality of the cigarettes. The above results suggest that FTIR microspectroscopy can identify different grades of the cigarettes, which may be helpful for tobacco research.

Electrodialysis Metathesis (EDM) Desalination for the Effective Removal of Chloride and Nitrate from Tobacco Extract: The Effect of Membrane Type.

Electrodialysis Metathesis (EDM) desalination was investigated using a squad of three ion-exchange membranes (ACS, TW-A, and A3) and simulated tobacco extract liquid for selective ions removal. We have studied various factors affecting EDM desalination efficiency using a complete experimental design. First, diffusion dialysis (DD) was conducted to determine the permeation rate of different anions in tobacco liquor with different membrane materials. We conclude that A3 had the fastest permeation rate of anions. However, ACS has the lowest permeation rate for different salts. The investigation of the EDM process showed the excellent ion permeation ability of A3 by detecting the current, conductivity, and ion concentration of the target tobacco liquor in the metathesis chamber of the EDM process. The EDM had shown the most excellent chloride ion removal ability. We found that A3 was the best membrane for the EDM process of tobacco liquor.

A novel approach for quantitative determination of cellulose content in tobacco via 2D HSQC NMR spectroscopy.

Cellulose is an important component of tobacco (Nicotiana tabacum L.) cell walls, which can be precursors for many harmful compounds in smoke. Traditional cellulose content analysis methods involve sequential extraction and separation steps, which are time-consuming and environmentally unfriendly. In this study, a novel method was first introduced to analyze cellulose content in tobacco via two-dimensional heteronuclear single quantum coherence (2D HSQC) NMR spectroscopy. The method was based on derivatization approach to allow the dissolution of insoluble polysaccharide fractions of tobacco cell walls in DMSO­d6/pyridine-d5 (4:1 v/v) for NMR analysis. The NMR results suggested that besides the main NMR signals of cellulose, partial signals of hemicellulose including mannopyranose, arabinofuranose, and galactopyranose units could also be identified. In addition, the utilization of relaxation reagents has proved to be an effective way to improve the sensitivity of 2D NMR spectroscopy, which was beneficial for quantification of biological samples with limited quantities. To overcome the limitations of quantification using 2D NMR, the calibration curve of cellulose with 1,3,5-trimethoxybenzene as internal reference was constructed and thus the accurate measurement of cellulose in tobacco was achieved. Compared with the chemical method, the interesting method was simple, reliable, and environmentally friendly, which provided a new insight for quantitative determination and structure analysis of plant macromolecules in complex samples.

Expression of bacterial GshF in Pichia pastoris for glutathione production.

Conventionally, two consecutive enzymatic reactions catalyzed by γ-glutamylcysteine synthetase and glutathione synthetase are most commonly used for glutathione production. Here we demonstrate that bacterial bifunctional GshF can be used for glutathione production in a eukaryotic system without accumulation of the intermediate γ-glutamylcysteine.

Plasma Treatment for Cellulose in Tobacco Paper-Base: The Improvement of Surface Hydrophilicity and Mechanical Property.

This paper reports a plasma treatment (PT) method for improving the surface hydrophilicity and mechanical properties of cellulose in reconstituted tobacco paper-base. The absorption and infiltration rates of water droplets on PT-reconstituted tobacco paper-base-15 s were significantly accelerated. Notably, the increased content of methylene and alkyl groups enabled the tobacco paper-base to absorb more useful substrates in the tobacco extract after plasma treatment. In addition, the tensile mechanical performance of reconstituted tobacco was significantly improved after plasma treatment, which indicated that the content of organic matter absorbed by the tobacco paper-base sheet was increased. Moreover, tobacco extract infiltrated on the surface of PT-reconstituted tobacco paper-base reached 37.7° within 30 s, while it reached 79.9° on the reconstituted tobacco paper-base. Finally, the mechanism by which the surface hydrophilicity and mechanical properties of the cellulose in the tobacco paper-base were improved is discussed.

A Comprehensive Analysis of Inorganic Ions and Their Selective Removal from the Reconstituted Tobacco Extract Using Electrodialysis.

Many tobacco stalks, dust, and fines are discharged in the tobacco industry, rich in inorganic minerals ions and nicotine salts. The high salinity and nicotine salts are challenging to be addressed by traditional treatment and are a severe threat that ought to be overcome. Thus, proper techniques can regenerate the tobacco stalks into reconstituted tobacco flakes used as cigarette filler. The electrodialysis process has been a viable approach to removing the inorganic ingredients in wastewater. We studied concentration, pH, and co-related influences with the nicotine and sugar/nicotine contents on the desalination performance. The results show that the inorganic ions such as Cl-, K+, Ca2+, and Mg2+ ions were successfully removed. When the feed concentration ranges from 3 to 15%, the removal ratio of the K+ ions is higher than Ca2+ and Mg2+ ions. As we reported previously, the K+ and Ca2+ ions are unfavorable for the total particulate matter emission but beneficial to decreasing the HCN delivery in mainstream cigarette smoke. Selective ED is a robust technology to reduce the harmful component delivery in cigarette smoke.

Electrodialytic Desalination of Tobacco Sheet Extract: Membrane Fouling Mechanism and Mitigation Strategies.

In the papermaking industry (reconstituted tobacco), a large number of tobacco stems, dust, and fines are discharged in the wastewater. This high salinity wastewater rich in ionic constituents and nicotine is difficult to be degraded by conventional biological treatment and is a serious threat that needs to be overcome. Electrodialysis (ED) has proved a feasible technique to remove the inorganic components in the papermaking wastewater. However, the fouling in ion exchange membranes causes deterioration of membranes, which causes a decrease in the flux and an increase in the electrical resistance of the membranes. In this study, the fouling potential of the membranes was analyzed by comparing the properties of the pristine and fouled ion exchange membranes. The physical and chemical properties of the ion exchange membranes were investigated in terms of electrical resistance, water content, and ion exchange capacity, as well as studied by infrared spectroscopy (IR) spectra, scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) analyses. The results indicated that the membrane fouling is caused by two different mechanisms. For the anion exchange membranes, the fouling is mainly caused by the charged organic anions. For the cation exchange membrane, the fouling is caused by minerals such as Ca2+ and Mg2+. These metal ions reacted with OH- ions generated by water dissociation and precipitated on the membrane surface. The chemical cleaning with alkaline and acid could mitigate the fouling potential of the ion exchange membranes.

A High-Throughput, Multi-Index Isothermal Amplification Platform for Rapid Detection of 19 Types of Common Respiratory Viruses Including SARS-CoV-2.

Fast and accurate diagnosis and the immediate isolation of patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are regarded as the most effective measures to restrain the coronavirus disease 2019 (COVID-19) pandemic. Here, we present a high-throughput, multi-index nucleic acid isothermal amplification analyzer (RTisochip™-W) employing a centrifugal microfluidic chip to detect 19 common respiratory viruses, including SARS-CoV-2, from 16 samples in a single run within 90 min. The limits of detection of all the viruses analyzed by the RTisochip™-W system were equal to or less than 50 copies·µL-1, which is comparable to those of conventional reverse transcription polymerase chain reaction. We also demonstrate that the RTisochip™-W system possesses the advantages of good repeatability, strong robustness, and high specificity. Finally, we analyzed 201 cases of preclinical samples, 14 cases of COVID-19-positive samples, 25 cases of clinically diagnosed samples, and 614 cases of clinical samples from patients or suspected patients with respiratory tract infections using the RTisochip™-W system. The test results matched the referenced results well and reflected the epidemic characteristics of the respiratory infectious diseases. The coincidence rate of the RTisochip™-W with the referenced kits was 98.15% for the detection of SARS-CoV-2. Based on these extensive trials, we believe that the RTisochip™-W system provides a powerful platform for fighting the COVID-19 pandemic.

Development of a rapid method for the quantification of cellulose in tobacco by (13)C CP/MAS NMR.

A method was developed for rapid quantitative determination of cellulose in tobacco by utilizing (13)C cross polarization magic angle spinning NMR spectroscopy ((13)C CP/MAS NMR). Sample powder was loaded into NMR rotor, which was customized rotor containing a matched silicon tube as an intensity reference. (13)C CP/MAS NMR spectra of tobacco samples were processed with spectral deconvolution to obtain the area of the C-1 resonance at 105.5ppm and the internal standard at 0ppm. The ratio between the area of 105.5ppm and 0ppm of a set of standard cellulose samples was used to construct a calibration curve. The cellulose content of a tobacco sample was determined by comparison of the ratio between the area of 105.5ppm and 0ppm to the calibration curve. Results of this developed method showed good agreement with those obtained from chemical analysis. The proposed method has such advantages of accuracy, quickness and efficiency, and could be an alternative to chemical analyses of cellulose.

Beta-Aminobutyric acid-mediated enhancement of resistance in tobacco against TMV and consideration of its capability in wounded tobacco plants.

Systemic acquired resistance (SAR) is an important component of disease-resistance arsenal of plants, and is associated with enhanced potency of activating local and systemic defense-related responses upon pathogen attack. In this report, we demonstrated that pre-treatment with beta-aminobutyric acid (BABA), a new elicitor of SAR in the plants, enhanced resistance against tobacco mosaic virus (TMV) in a temperately-sensitive tobacco (Nicotiana tabacum L.) cultivar Yunyan 85. The resistance is based on the elicitation of defense-related responses induced by BABA that brings the TMV-susceptible tobacco plants to a defense-ready state, even before exposure to the pathogen. The induced resistance was strongly associated with potentiated activation of defense-related enzymes [phenylalanine ammonia lyase (PAL) and polyphenol oxidase (PPO)] activities, proportional to the concentration of the BABA sprayed. Interestingly, simultaneous clipping, an important agricultural practice in tobacco production, attenuated BABA-mediated enhancement of TMV resistance in tobacco. The changes in the defense-related enzymes activities indicated that the interaction between BABA and wounding was reciprocally antagonistic. Moreover, such a negative interaction regulated the expression of defense-related enzymes. depending on the time of induction.