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1.
J Chromatogr A ; 1626: 461361, 2020 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-32797840

RESUMO

The alkaloid enantiomers are well-known to have different physiological and pharmacological effects, and to play an important role in enantioselectivity metabolism with enzymes catalysis in tobacco plants. Here, we developed an improved method for simultaneous and high-precision determination of the individual enantiomers of nornicotine, anatabine and anabasine in four tobacco matrices, based on an achiral gas chromatography-nitrogen phosphorus detector (GCNPD) with commonly available Rtx-200 column using (1S)-(-)-camphanic chloride derivatization. The method development consists of the optimization of extraction and derivatization, screening of achiral column, analysis of the fragmentation mechanisms and evaluation of matrix effect (ME). Under the optimized experimental conditions, the current method exhibited excellent detection capability for the alkaloid enantiomers, with coefficients of determination (R2) > 0.9989 and normality test of residuals P > 0.05 in linear regression parameters. The ME can be neglected for the camphanic derivatives. The limit of detection (LOD) and limit of quantitation (LOQ) ranged from 0.087 to 0.24 µg g - 1 and 0.29 to 0.81 µg g - 1, respectively. The recoveries and within-laboratory relative standard deviations (RSDR) were 94.3%~104.2% and 0.51%~3.89%, respectively. The developed method was successfully applied to determine the enantiomeric profiling of cultivars and curing processes. Tobacco cultivars had a significant impact on the nornicotine, anatabine, anabasine concentration and enantiomeric fraction (EF) of (R)-nornicotine, whereas the only significant change induced by the curing processes was an increase in the EF of (R)-anabasine.


Assuntos
Alcaloides/análise , Anabasina/análise , Cromatografia Gasosa/métodos , Nicotina/análogos & derivados , Piridinas/análise , Tabaco/química , Alcaloides/química , Anabasina/química , Hidrocarbonetos Aromáticos com Pontes/química , Cloretos/química , Lactonas/química , Nicotina/análise , Nicotina/química , Piridinas/química , Estereoisomerismo
2.
Exp Parasitol ; 217: 107955, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32649953

RESUMO

Nicotiana tabacum, Stemona japonica, and Cnidium monnieri are common plants that are widely used for their anti-parasitic properties. The purpose of this study was to evaluate the acaricidal activity of extracts from these plants against the brown dog tick, Rhipicephalus sanguineus. A composition analysis of crude extracts by GC-MS was conducted to discover compounds with acaricidal effects. The toxicity of extraction against the engorged nymphs of R. sanguineus was evaluated by an immersion test. The results showed that the crude extracts of S. japonica and C. monnieri in varying ratios, concentrations, and from different extraction methods, had a killing effect on R. sanguineus. Lethality reached 76.67% ± 0.04410 when using a 1:1 extract of S. japonica:C. monnieri in 75% ethanol with ultrasonic extraction; the crude extract was determined at a concentration of 0.5 g/mL. GC-MS results showed that osthole and 5-hydroxymethylfurfural (5-HMF) are the main components of the extract. These results suggested that ultrasound-assisted extraction (UAE) extracts contained acaricidal components acting against R. sanguineus, which may result in the development of effective extracts of S. japonica and C. monnieri as a source of low-toxicity, plant-based, natural acaricidal drugs.


Assuntos
Cnidium/química , Extratos Vegetais/farmacologia , Rhipicephalus sanguineus/efeitos dos fármacos , Stemonaceae/química , Controle de Ácaros e Carrapatos/métodos , Animais , Bioensaio , Cumarínicos/análise , Cumarínicos/farmacologia , Furaldeído/análogos & derivados , Furaldeído/análise , Furaldeído/farmacologia , Cromatografia Gasosa-Espectrometria de Massas , Muda/efeitos dos fármacos , Ninfa/efeitos dos fármacos , Extratos Vegetais/química , Coelhos , Tabaco/química
3.
PLoS One ; 15(5): e0232102, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32437367

RESUMO

Cigarette smoke exposure is a risk factor for many pulmonary diseases, including Chronic Obstructive Pulmonary Disease (COPD). Cigarette smokers are more prone to respiratory infections with more severe symptoms. In those with COPD, viral infections can lead to acute exacerbations resulting in lung function decline and death. Epithelial cells in the lung are the first line of defense against inhaled insults such as tobacco smoke and are the target for many respiratory pathogens. Endocytosis is an essential cell function involved in nutrient uptake, cell signaling, and sensing of the extracellular environment, yet, the effect of cigarette smoke on epithelial cell endocytosis is not known. Here, we report for the first time that cigarette smoke alters the function of several important endocytic pathways in primary human small airway epithelial cells. Cigarette smoke exposure impairs clathrin-mediated endocytosis and fluid phase macropinocytosis while increasing caveolin mediated endocytosis. We also show that influenza virus uptake is enhanced by cigarette smoke exposure. These results support the concept that cigarette smoke-induced dysregulation of endocytosis contributes to lung infection in smokers. Targeting endocytosis pathways to restore normal epithelial cell function may be a new therapeutic approach to reduce respiratory infections in current and former smokers.


Assuntos
Caveolinas/metabolismo , Células Epiteliais/efeitos dos fármacos , Infecções/patologia , Pulmão/citologia , Fumaça/efeitos adversos , Tabaco/química , Regulação para Cima/efeitos dos fármacos , Suscetibilidade a Doenças , Células Epiteliais/metabolismo , Células Epiteliais/virologia , Humanos , Infecções/induzido quimicamente , Infecções/virologia , Tabaco/efeitos adversos
4.
PLoS One ; 15(5): e0233111, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32453764

RESUMO

Bangladesh exhibits the second highest rate of smokeless tobacco (SLT) product usage in the world, and this has been associated with the high upper aerodigestive tract cancer incidence in this country. The goal of the present study was to examine the levels of the highly carcinogenic tobacco-specific nitrosamines (TSNAs) in Bangladeshi SLT products and compare these levels to that observed in SLT brands from southeast Asia and the USA. The levels of TSNAs and nicotine were determined by LC-MS/MS in twenty-eight SLT brands and several tobacco additives from Bangladesh, as well as several SLT brands from India, Pakistan and the USA. The levels of N-nitrosonornicotine (NNN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), N-nitrosoanatabine (NAT) and N-nitrosoanabasine (NAB) in Bangladeshi SLT brands ranged from 1.1-59, 0.15-34, 0.79-45, and 0.037-13 µg/g SLT powder, respectively. The mean levels of the highly carcinogenic TSNAs (NNN+NNK) were 7.4-, 2.4-, and 63-fold higher in Bangladeshi SLT products as compared to SLT brands from the USA, India and Pakistan, respectively; these trends were also observed for NAT and NAB. Similar mean levels of nicotine were observed in the Bangladeshi brands (31 mg/g powder) versus brands from the USA (25 mg/g powder) and India (20 mg/g powder); they were 3-fold higher than brands from Pakistan (10 mg/g powder). Gul SLT brands exhibited the highest pH and the highest levels of unprotonated nicotine. The high levels of TSNAs in Bangladeshi SLT brands may be an important factor contributing to the high rates of upper aerodigestive tract cancer in Bangladesh.


Assuntos
Nitrosaminas/análise , Tabaco sem Fumaça/análise , Tabaco/química , Bangladesh , Cromatografia Líquida , Nicotina/análise , Espectrometria de Massas em Tandem , Produtos do Tabaco/análise
5.
Toxicol Lett ; 331: 57-64, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32442718

RESUMO

Electrophilic compounds present in humans, originating from endogenous processes or pollutant exposures, pose a risk to health though their reaction with nucleophilic sites in protein and DNA. Among this chemical class, aldehydes are mainly present in indoor air and they can also be produced by endogenous lipid peroxidation arising from oxidative stress. Known to be very reactive, aldehydes have the ability to form exocyclic adducts to DNA that, for the most if not repaired correctly, are mutagenic and by consequence potential agents involved in carcinogenesis. The aim of this work was to establish profiles of exocyclic DNA adducts induced by aldehyde mixtures, which could ultimately be considered as a genotoxic marker of endogenous and environmental aldehyde exposure. Adducts were quantified by an accurate, sensitive and validated ultra high performance liquid chromatography-electrospray ionization analytical method coupled to mass spectrometry in the tandem mode (UHPLC-ESI-MS/MS). We simultaneously measured nine exocyclic DNA adducts generated during the exposure in vitro of calf thymus DNA to different concentrations of each aldehyde along, as well as, to an equimolar mixture of these aldehydes. This approach has enabled us to establish dose-response relationships that allowed displaying the specific reactivity of aldehydes towards corresponding adducts formation. Profiles of these adducts determined in DNA of current smokers and non-smokers blood samples supported these findings. These first results are encouraging to explore genotoxicity induced by aldehyde mixtures and can furthermore be used as future reference for adductomic approaches.


Assuntos
Aldeídos/toxicidade , Adutos de DNA/sangue , DNA/efeitos dos fármacos , Mutagênicos/toxicidade , Fumar/sangue , Tabaco , DNA/genética , Relação Dose-Resposta a Droga , Humanos , Tabaco/química
6.
Ecotoxicol Environ Saf ; 197: 110603, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32304920

RESUMO

Both sides of tobacco leaves accumulate 210Po through their sticky hairs (trichomes) by means of diffusive deposition. It is known that tobacco leaves may contain high amounts of 210Po concentrations. However, there is less information about radionuclides in tobacco plants. In the study, the 210Po concentrations were determined monthly in different morphological parts of tobacco plants (leaf, stem, root) and soil samples and soil-to-plant transfer factor for 210Po was determined and the activity concentrations of 210Po radionuclide in 16 different popular brands of cigarettes were investigated in order to estimate the annual effective doses of 210Po to smokers. Besides the study investigated (focused on) the correlation between the amount of rainfall and 210Po concentration of tobacco leaf. The results of the present study indicated that the activity concentration of 210Po in cigarettes distributed in Turkey ranged from 16.1 ± 1.0 to 37.6 ± 2.6 mBq per cigarette and the mean value of their activity concentrations was 22.4 ± 1.5 mBq per cigarette. The mean value of annual effective doses of 210Po to smokers (20 cigarettes smoked by an individual per day) obtained from these activity concentrations was estimated to be 188.5 ± 12.4 µSv y-1.


Assuntos
Polônio/análise , Produtos do Tabaco/análise , Tabaco/química , Humanos , Folhas de Planta/química , Raízes de Plantas/química , Caules de Planta/química , Doses de Radiação , Fumantes , Solo/química , Turquia
7.
J Toxicol Sci ; 45(3): 177-186, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32147640

RESUMO

MECP2 (Methyl-CpG-binding protein 2) has been shown to have a critical role in regulating DNA methylation against smoke exposed lung injury. However, the biological function of MECP2 and the underlying molecular mechanism remains elusive. Human bronchial epithelial (16HBE) and alveolar type II epithelial cells (AECII) were exposed to increasing concentrations of cigarette smoke extracts (CSE) solution to establish CSE-induced lung epithelial cell injury models. Our findings revealed that MECP2 was down-regulated, while CYP1B1 was up-regulated in CSE-induced lung epithelial cell injury models by quantitative real time PCR, western blotting and immunofluorescence staining. Down-regulated CYP1B1 was ascribed to the demethylation of its promoter by methylation-specific PCR (MSP). The in vitro experiments further showed that MECP2 overexpression significantly attenuated CSE-triggered cell growth attenuation, cell cycle arrest, apoptosis and ROS generation in lung epithelial cells by CCK-8 and flow cytometry assays. In molecular level, we further demonstrated that MECP2 overexpression obviously suppressed the expression of CYP1B1 through enhancing DNA methylation. Therefore, our data suggest that MECP2 protects against CSE-induced lung epithelial cell injury possibly through down-regulating CYP1B1 expression via elevating its methylation status.


Assuntos
Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/genética , Fumar Cigarros/efeitos adversos , Citocromo P-450 CYP1B1/metabolismo , Metilação de DNA , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Expressão Gênica , Pulmão/citologia , Proteína 2 de Ligação a Metil-CpG/genética , Proteína 2 de Ligação a Metil-CpG/metabolismo , Extratos Vegetais/efeitos adversos , Tabaco/química , Lesão Pulmonar Aguda/patologia , Células Cultivadas , Citocromo P-450 CYP1B1/genética , Humanos , Extratos Vegetais/isolamento & purificação
8.
PLoS One ; 15(2): e0229256, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32084204

RESUMO

Cigarette smoke (CS) contains multiple gaseous and particulate materials that can cause lung inflammation, and smoking is the major cause of chronic obstructive pulmonary disease (COPD). We sought to determine the mechanisms of how CS triggers lung inflammation. Nur77, a nuclear hormone receptor belonging to the immediate-early response gene family, controls inflammatory responses, mainly by suppressing the NF-κB signaling pathway. Because it is unknown if Nur77's anti-inflammatory role modulates COPD, we assessed if and how Nur77 expression and activity are altered in CS-induced airway inflammation. In lung tissues and bronchial epithelial cells from COPD patients, we found Nur77 was downregulated. In a murine model of CS-induced airway inflammation, CS promoted lung inflammation and also reduced Nur77 activity in wild type (WT) mice, whereas lungs of Nur77-deficient mice showed exaggerated CS-induced inflammatory responses. Our findings in in vitro studies of human airway epithelial cells complemented those in vivo data in mice, together showing that CS induced threonine-phosphorylation of Nur77, which is known to interfere with its anti-inflammatory functions. In summary, our findings point to Nur77 as an important regulator of CS-induced inflammatory responses and support the potential benefits of Nur77 activation for COPD treatment.


Assuntos
Regulação para Baixo/efeitos dos fármacos , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Doença Pulmonar Obstrutiva Crônica/genética , Fumaça/efeitos adversos , Tabaco/química , Animais , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Humanos , Inflamação/genética , Pulmão/patologia , Camundongos , Fosforilação/efeitos dos fármacos , Doença Pulmonar Obstrutiva Crônica/patologia , Treonina/metabolismo
9.
J Anal Toxicol ; 44(5): 514-520, 2020 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-31984423

RESUMO

Smoking is a public health problem and an important source of exposure to toxic metals. This work describes an efficient analytical method comparable to the ones based on atomic emission techniques for the determination of chromium in different constituent parts of cigarette samples (tobacco, filters and ashes) using electrothermal vaporization-atomic absorption spectrometry. The method was evaluated using 12 samples, and the results showed recovery values between 83 and 107%. The accuracy was also evaluated using a reference sample of tomato leaves (NIST SRM 1573a), which proved the efficiency of the method. The limits of detection of the developed method were 20.4, 75.8 and 80.7 ng g-1 for tobacco, filter and cigarette ash samples, respectively. The average chromium values found for the analyzed samples were in the range of 0.96 to 3.85 and from 0.32 to 0.80 µg/cigarette for tobacco and ashes, respectively. For most pre-burn and post-burn filter samples, the values of chromium concentration remained below limits of detection. The developed method presented adequate results about precision and accuracy, demonstrating its applicability in the determination of chromium in cigarette samples.


Assuntos
Cromo/análise , Produtos do Tabaco/análise , Brasil , Humanos , Fumar , Tabaco/química , Produtos do Tabaco/legislação & jurisprudência
10.
Plant Biol (Stuttg) ; 22(1): 70-80, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31283085

RESUMO

The effects of elevated glutathione levels on defence responses to powdery mildew (Euoidium longipes) were investigated in a salicylic acid-deficient tobacco (Nicotiana tabacum cv. Xanthi NahG) and wild-type cv. Xanthi plants, where salicylic acid (SA) contents are normal. Aqueous solutions of reduced glutathione (GSH) and its synthetic precursor R-2-oxothiazolidine-4-carboxylic acid (OTC) were injected into leaves of tobacco plants 3 h before powdery mildew inoculation. SA-deficient NahG tobacco was hyper-susceptible to E. longipes, as judged by significantly more severe powdery mildew symptoms and enhanced pathogen accumulation. Strikingly, elevation of GSH levels in SA-deficient NahG tobacco restored susceptibility to E. longipes to the extent seen in wild-type plants (i.e. enhanced basal resistance). However, expression of the SA-mediated pathogenesis-related gene (NtPR-1a) did not increase significantly in GSH or OTC-pretreated and powdery mildew-inoculated NahG tobacco, suggesting that the induction of this PR gene may not be directly involved in the defence responses induced by GSH. Our results demonstrate that artificial elevation of glutathione content can significantly reduce susceptibility to powdery mildew in SA-deficient tobacco.


Assuntos
Ascomicetos , Resistência à Doença , Glutationa , Ácido Salicílico , Tabaco , Ascomicetos/efeitos dos fármacos , Ascomicetos/fisiologia , Resistência à Doença/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas , Glutationa/metabolismo , Glutationa/farmacologia , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/microbiologia , Tabaco/química , Tabaco/microbiologia
11.
Environ Pollut ; 259: 113841, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31883477

RESUMO

Nicotine (Nic) exposed to the environment which comes from tobacco products is the main addictive agent and specific classes of hazardous compound that merit concern. In this study, we have established a fast and reliable method to achieve specific detection of Nic in natural nicotiana tabacum within 30 s through a miniaturized platform based on screen printed gold electrode (SPE). A simple electrochemical pretreatment mean was employed on gold surface that led to the exposure of Au (111) facet and a convenient sample pretreatment method was adopted to realize the extraction of Nic in tobacco. The present electrochemical sensor exhibits an ample range of sensing from 10 µg/g to 200 µg/g, which is able to compliance with tobacco industry testing standards of actual samples. Over 60 sampling points from different origins in China or other countries were performed with direct analysis using this method and satisfactory results have been obtained. The proposed approach was demonstrated to be a very promising platform for significantly improving analytical efficiency in laboratories as well as for monitoring the source reduction control of Nic in the environment.


Assuntos
Técnicas de Química Analítica/métodos , Nicotina , Tabaco , China , Eletrodos , Nicotina/análise , Tabaco/química
12.
Phytochemistry ; 170: 112187, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31865001

RESUMO

Nicotinic acetylcholine receptor (nAChR) subtype-selective pharmacological profiles of tobacco alkaloids are essential for understanding the physiological effects of tobacco products. In this study, automated electrophysiology was used to functionally characterize the effects of distinct groups of tobacco alkaloids on human α4ß2 and α7 nAChRs. We found that, in tobacco alkaloids, pyridine as a hydrogen bond acceptor and a basic nitrogen atom at a distance of 4-7 Šare pharmacophoric elements necessary for molecular recognition by α4ß2 and α7 nAChRs with various degrees of selectivity, potency, and efficacy. While four alkaloids-nicotine, nornicotine, anabasine and R-anatabine-potently activated α4ß2, they were also weak agonists of α7 nAChRs. Nicotine was the most potent agonist of α4ß2, while anabasine elicited the highest activation of α7. None of the tobacco alkaloids enhanced nAChR activity elicited by the endogenous ligand acetylcholine; therefore, none was considered to be a positive allosteric modulator (PAM) of either α4ß2 or α7 nAChRs. In contrast, we identified tobacco alkaloids, such as the tryptophan metabolite 6-hydroxykynurenic acid, that decreased the activity of both α4ß2 and α7 nAChRs. Our study identified a class of alkaloids with positive and negative effects against human α4ß2 and α7 nAChRs. It also revealed human α4ß2 to be the principal receptor for sensing the most abundant alkaloids in tobacco leaves.


Assuntos
Alcaloides/farmacologia , Produtos Biológicos/farmacologia , Compostos Fitoquímicos/farmacologia , Receptores Nicotínicos/metabolismo , Tabaco/química , Receptor Nicotínico de Acetilcolina alfa7/agonistas , Alcaloides/química , Alcaloides/isolamento & purificação , Produtos Biológicos/química , Produtos Biológicos/isolamento & purificação , Relação Dose-Resposta a Droga , Humanos , Ligantes , Estrutura Molecular , Compostos Fitoquímicos/química , Compostos Fitoquímicos/isolamento & purificação , Relação Estrutura-Atividade , Receptor Nicotínico de Acetilcolina alfa7/metabolismo
13.
BMC Biotechnol ; 19(1): 101, 2019 12 21.
Artigo em Inglês | MEDLINE | ID: mdl-31864319

RESUMO

BACKGROUND: Clostridium perfringens, a gram-positive, anaerobic, rod-shaped bacterium, is the third leading cause of human foodborne bacterial disease and a cause of necrotic enteritis in poultry. It is controlled using antibiotics, widespread use of which may lead to development of drug-resistant bacteria. Bacteriophage-encoded endolysins that degrade peptidoglycans in the bacterial cell wall are potential replacements for antibiotics. Phage endolysins have been identified that exhibit antibacterial activities against several Clostridium strains. RESULTS: An Escherichia coli codon-optimized gene encoding the glycosyl hydrolase endolysin (PlyCP41) containing a polyhistidine tag was expressed in E. coli. In addition, The E. coli optimized endolysin gene was engineered for expression in plants (PlyCP41p) and a plant codon-optimized gene (PlyCP41pc), both containing a polyhistidine tag, were expressed in Nicotiana benthamiana plants using a potato virus X (PVX)-based transient expression vector. PlyCP41p accumulated to ~ 1% total soluble protein (100µg/gm f. wt. leaf tissue) without any obvious toxic effects on plant cells, and both the purified protein and plant sap containing the protein lysed C. perfringens strain Cp39 in a plate lysis assay. Optimal systemic expression of PlyCP41p was achieved at 2 weeks-post-infection. PlyCP41pc did not accumulate to higher levels than PlyCP41p in infected tissue. CONCLUSION: We demonstrated that functionally active bacteriophage PlyCP41 endolysin can be produced in systemically infected plant tissue with potential for use of crude plant sap as an effective antimicrobial agent against C. perfringens.


Assuntos
Bacteriófagos/enzimologia , Clostridium perfringens/efeitos dos fármacos , Endopeptidases/genética , Tabaco/genética , Proteínas Virais/genética , Bacteriófagos/genética , Clostridium perfringens/fisiologia , Endopeptidases/química , Endopeptidases/metabolismo , Endopeptidases/farmacologia , Expressão Gênica , Folhas de Planta/química , Folhas de Planta/genética , Folhas de Planta/metabolismo , Engenharia de Proteínas , Tabaco/química , Tabaco/metabolismo , Proteínas Virais/química , Proteínas Virais/metabolismo , Proteínas Virais/farmacologia
14.
Molecules ; 25(1)2019 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-31861797

RESUMO

N. glutinosa L. is a relatively less studied Nicotiana species (Solanaceae), although there are data about its importance as a model plant in viral control studies, as a gene donor in tobacco hybridization and as a source of agents with insecticidal or fungicidal effects. The biological activities of the species were associated mostly with the presence of leaf surface metabolites, in particular diterpenes and sucrose esters. The aim of this study was to identify the chemical composition of the essential oil (EO) and two aromatic extraction products (concrete and resinoid) obtained from N. glutinosa L. leaves. GC-MS analysis identified 26 components in the EO (representing 97.3% of total oil content), which contained mostly diterpene compounds with major components manool (14.2%), sclarene (8.4%) and manoyl oxide (8.1%). The number of compounds identified in the concrete was 37 (95.5% of the total content) and the major component was the diterpene alcohol sclareol (14.2%). In the resinoid, 30 volatile components (representing 95.1% of resinoid content) were identified, with major components nicotine (32.9%), α-tocopherol (8.2%), tridecanoin (6.9%), sclareol (6.9%), and solanone (6.9%). The group of bicyclic diterpenes had the largest share in the diterpene fraction of the products (57.3%, 91.7%, and 86.3%, respectively for the EO, concrete, and resinoid). Considering the abundance of sclareol in the aromatic products, the antimicrobial activity of the pure substance was determined. Sclareol was highly effective against a set of medicinally important yeasts; Candida albicans АТСС 10231, C. glabrata ATCC 90030, C. parapsilosis clinical isolate, and C. tropicalis NBIMCC 23, while being less effective against the studied Gram-positive and Gram-negative bacteria. Data from the study on N. glutinosa aromatic products composition may be of interest to the aroma industries for their possible use in perfumery and cosmetics.


Assuntos
Anti-Infecciosos , Candida albicans/crescimento & desenvolvimento , Bactérias Gram-Negativas/crescimento & desenvolvimento , Bactérias Gram-Positivas/crescimento & desenvolvimento , Óleos Voláteis , Folhas de Planta/química , Terpenos , Tabaco/química , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Terpenos/química , Terpenos/farmacologia
15.
BMC Plant Biol ; 19(1): 555, 2019 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-31842767

RESUMO

BACKGROUND: The leaf age for harvesting flue-cured tobacco leaves is closely related to the quality of tobacco leaves, so an appropriate leaf age for harvesting is important for improving yield and quality of flue-cured tobacco, however, at present, there are few studies on effects of leaf age on physiological and biochemical changes during flue-curing and there is no clear standard of proper leaf ages for harvesting in production. RESULTS: In the Yunnan tobacco-growing area, an experiment was carried from 2016 to 2017 and different leaf ages were set. The results demonstrate that leaf age has a significant on tissue cell gap, leaf age and flue-curing stages exert significant effects on upper epidermis, palisade and spongy tissue, and leaf thickness of tobacco leaves. The thicknesses of upper and lower epidermis as well as palisade and spongy tissues at different ages show an approximately W-shaped change trend during flue-curing. With the advance of flue-curing stages, contents of starch, chlorophyll, carotenoid, and water in tobacco leaves at different leaf ages decrease, while polyphenol and malondialdehyde (MDA) contents increase. The older the leaf, the faster the chlorophyll, carotenoid, and water contents reduce, while the faster the polyphenol and MDA content rise during flue-curing. The flue-cured tobacco leaves at 116 DAT (days after transplanting) show the highest contents of total nitrogen and nicotine, followed by 123 DAT and those at 130 DAT are the lowest; however, the contents of total sugar and reducing sugar demonstrate a contrary tendency, and the starch content at 116 DAT is much lower than those in the other two treatments. The proportion of superior tobacco, average price, yield, and output value of upper tobacco leaves at different leaf ages are the highest at 123 DAT. The highest sensory evaluation score is found at 123 DAT, while that at 130 DAT is significantly lower in comparison with the other two treatments. CONCLUSIONS: Tobacco leaves harvested at 123 DAT are mature and exhibit a low degree of membrane lipid peroxidation, moderate chemical compositions, and high economic value. 123 DAT improves availability of tobacco leaves.


Assuntos
Folhas de Planta/química , Tabaco/química , Fatores de Tempo
16.
Molecules ; 24(24)2019 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-31861109

RESUMO

The content of tobacco-specific nitrosamines (TSNAs) possessing carcinogenic properties has been an important area of research since replacement liquids were introduced for e-cigarettes. A method for determining N'-nitrosonornicotine (NNN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), N'-nitrosoanatabine (NAT), and N'-nitrosoanabasine (NAB) in replacement liquids for electronic cigarettes was developed using liquid chromatography-tandem mass spectrometry with electrospray ionisation (HPLC-ESI-MS/MS) in the multiple reaction monitoring mode. The sample preparation of replacement liquids was accomplished via the ultrasound-assisted solvent extraction of a porous membrane packed sample. The sample preparation proved to be successful in extracting the analytes, with recoveries from 87% to 105%, with coefficients of variation < 4.9%. Moreover, the linearity and limits of detection and quantitation (LOD, LOQ), together with repeatability and accuracy, were determined for the developed method. The proposed sample preparation and developed chromatographic method were successfully applied to the determination of TSNAs in 9 replacement liquid samples. The NNK and NNN were found to be most frequently detected (89 and 67%, respectively), with concentration ranges from 1.2-54.3 ng/mL and 4.1-30.2 ng/mL, respectively, while NAT was detected with frequency of 22% with range 1.7-2.5 ng/mL and NAB were found to be below the LOD in all samples.


Assuntos
Fracionamento Químico , Sistemas Eletrônicos de Liberação de Nicotina , Membranas Artificiais , Nitrosaminas/análise , Solventes/análise , Tabaco/química , Ondas Ultrassônicas , Cromatografia Líquida , Nitrosaminas/isolamento & purificação , Porosidade , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem
17.
Mediators Inflamm ; 2019: 4693870, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31780859

RESUMO

Microbial lipopolysaccharides (LPS) have been implicated in the pathogenesis of rheumatoid arthritis (RA), possibly driving a systemic inflammatory response that may trigger the development and/or exacerbation of the disease. To explore the existence of this mechanism in African RA patients, we have measured systemic levels of LPS and its surrogate, LPS-binding protein (LBP), as well as those of intestinal fatty acid-binding protein (I-FABP), pulmonary surfactant protein D (SP-D), and cotinine in serum to identify possible origins of LPS, as well as associations of these biomarkers with rheumatoid factor (RF) and anticitrullinated peptide (aCCP) autoantibodies and the DAS 28-3 clinical disease severity score. A cohort of 40 disease-modifying antirheumatic drug-naïve, black South African RA patients rated by compound disease scores and 20 healthy subjects and 10 patients with chronic obstructive pulmonary disease (COPD) as controls were included in this study. Levels of the various biomarkers and autoantibodies were measured using a combination of ELISA and immunofluorimetric and immunoturbidometric procedures. LPS levels were lowest in the RA group compared to the healthy controls (p = 0.026) and COPD patients (p = 0.017), while LBP levels were also significantly lower in RA compared to the healthy individuals (p = 0.036). Levels of I-FABP and SP-D were comparable between all three groups. Categorisation of RA patients according to tobacco usage revealed the following significant positive correlations: LBP with C-reactive protein (p = 0.0137); a trend (p = 0.073) towards an association of LBP with the DAS 28-3 disease severity score; RF-IgG antibodies with both LPS and LBP (p = 0.033 and p = 0.041, respectively); aCCP-IgG antibodies with LPS (p = 0.044); and aCCP-IgG with RF-IgM autoantibodies (p = 0.0016). The findings of this study, several of them novel, imply that tobacco products, as opposed to microbial translocation, represent a potential source of LPS in this study cohort of RA patients, again underscoring the risks posed by tobacco usage for the development and severity of RA.


Assuntos
Artrite Reumatoide/induzido quimicamente , Lipopolissacarídeos/química , Artrite Reumatoide/sangue , Biomarcadores/sangue , Biomarcadores/metabolismo , Proteína C-Reativa/metabolismo , Ensaio de Imunoadsorção Enzimática , Proteínas de Ligação a Ácido Graxo/metabolismo , Feminino , Humanos , Masculino , Fator Reumatoide/metabolismo , Tabaco/química
18.
J Agric Food Chem ; 67(45): 12382-12392, 2019 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-31635461

RESUMO

Protoporphyrinogen oxidase (PPO, EC 1.3.3.4) is a promising target for herbicide discovery. Search for new compounds with novel chemotypes is a key objective for agrochemists. Here, we describe the discovery and systematic SAR-based structure optimization of novel N-isoxazolinylphenyltriazinones 5-9 as PPO inhibitors. The in vivo herbicidal activity and in vitro Nicotiana tabacum PPO (NtPPO) inhibitory activity were explored in detail. A number of the new synthetic compounds displayed strong PPO inhibitory activity with Ki values in the nanomolar range. Some compounds exhibited excellent and broad-spectrum weed control at the rate of 9.375-37.5 g ai/ha by postemergence application and showed improved monocotyledonous weed control compared to saflufenacil. Most promisingly, ethyl 3-(2-chloro-5-(3,5-dimethyl-2,6-dioxo-4-thioxo-1,3,5-triazinan-1-yl)-4-fluorophenyl)-5-methyl-4,5-dihydroisoxazole-5-carboxylate, 5a, with a Ki value of 4.9 nM, displayed over 2- and 6-fold higher potency than saflufenacil (Ki = 10 nM) and trifludimoxazin (Ki = 31 nM), respectively. Moreover, 5a showed excellent and broad-spectrum weed control against 32 kinds of weeds at 37.5-75 g ai/ha. Rice exhibited relative tolerance to 5a at 150 g ai/ha by postemergence application, indicating that 5a could be a potential herbicide candidate for weed control in paddy fields.


Assuntos
Inibidores Enzimáticos/farmacologia , Herbicidas/química , Herbicidas/farmacologia , Proteínas de Plantas/antagonistas & inibidores , Protoporfirinogênio Oxidase/antagonistas & inibidores , Descoberta de Drogas , Inibidores Enzimáticos/química , Cinética , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Daninhas/química , Plantas Daninhas/efeitos dos fármacos , Plantas Daninhas/enzimologia , Protoporfirinogênio Oxidase/química , Protoporfirinogênio Oxidase/metabolismo , Relação Quantitativa Estrutura-Atividade , Tabaco/química , Tabaco/efeitos dos fármacos , Tabaco/enzimologia , Controle de Plantas Daninhas
19.
Chem Biol Interact ; 314: 108846, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31606474

RESUMO

Matrix metalloproteinases (MMPs) have been implicated in EMT but their role in the regulation of cigarette smoke-induced EMT in airway epithelium is not clear. We have therefore investigated the potential role of MMP-2 and -9 in cigarette smoke extract (CSE) induced EMT using A549 lung epithelial cells and human small airway epithelial cells (SAEC). The cells were treated with different concentration of CSE, and MTT and trypan blue assays, acridine orange-ethidium bromide assay, gelatin zymography, Western blotting, immunofluorescence studies, Boyden-chamber assay, wound healing assay and air-liquid interface (ALI) culture were used to assess different cellular and molecular changes associated with EMT. The results depict that CSE increased the cytotoxicity along with a concurrent increase in the expression and activity of MMP-2 and -9. CSE further altered EMT markers like E-cadherin, N-cadherin, vimentin, and the molecular modulators of EMT such as ß-catenin and pGSK-3ß. Further, CSE also upregulated EGFR, AKT, and ERK1/2 in airway epithelial cells. SB-3CT, a known inhibitor of MMP-2 and -9, altered and reversed the expression of markers of EMT and kinases, validating the role of MMP-2 and -9 in CSE-induced EMT. Fisetin, a plant-derived bioflavonoid, also reversed the expression of EMT markers and molecular regulators in a similar fashion as SB-3CT. In summary, this study highlights the role of MMP-2 and -9 in CSE-induced EMT and curate its molecular cascade through EGFR/AKT/ERK/ß-catenin axis, which could be restored by MMP-2 and -9 inhibitor and fisetin. Fisetin is hitherto unknown to modulate CSE-induced MMPs activity in airway epithelial cells, and our study suggests its potential role as a therapeutic approach in CSE-induced EMT in lung epithelial cells.


Assuntos
Transição Epitelial-Mesenquimal/efeitos dos fármacos , Flavonoides/farmacologia , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fumaça/efeitos adversos , Tabaco/química , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Receptores ErbB/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Humanos , Proteínas Proto-Oncogênicas c-akt/metabolismo , beta Catenina/metabolismo
20.
Photosynth Res ; 142(3): 307-319, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31482263

RESUMO

A kinetic-LED-array-spectrophotometer (Klas) was recently developed for measuring in vivo redox changes of P700, plastocyanin (PCy), and ferredoxin (Fd) in the near-infrared (NIR). This spectrophotometer is used in the present work for in vitro light-induced measurements with various combinations of photosystem I (PSI) from tobacco and two different cyanobacteria, spinach plastocyanin, cyanobacterial cytochrome c6 (cyt. c6), and Fd. It is shown that cyt. c6 oxidation contributes to the NIR absorption changes. The reduction of (FAFB), the terminal electron acceptor of PSI, was also observed and the shape of the (FAFB) NIR difference spectrum is similar to that of Fd. The NIR difference spectra of the electron-transfer cofactors were compared between different organisms and to those previously measured in vivo, whereas the relative absorption coefficients of all cofactors were determined by using single PSI turnover conditions. Thus, the (840 nm minus 965 nm) extinction coefficients of the light-induced species (oxidized minus reduced for PC and cyt. c6, reduced minus oxidized for (FAFB), and Fd) were determined with values of 0.207 ± 0.004, - 0.033 ± 0.006, - 0.036 ± 0.008, and - 0.021 ± 0.005 for PCy, cyt. c6, (FAFB) (single reduction), and Fd, respectively, by taking a reference value of + 1 for P700+. The fact that the NIR P700 coefficient is larger than that of PCy and much larger than that of other contributing species, combined with the observed variability in the NIR P700 spectral shape, emphasizes that deconvolution of NIR signals into different components requires a very precise determination of the P700 spectrum.


Assuntos
Proteínas de Bactérias/química , Complexo de Proteína do Fotossistema I/metabolismo , Proteínas de Plantas/química , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Proteínas de Bactérias/metabolismo , Citocromos c6/química , Citocromos c6/metabolismo , Transporte de Elétrons , Ferredoxinas/metabolismo , Oxirredução , Complexo de Proteína do Fotossistema I/química , Proteínas de Plantas/metabolismo , Plastocianina/química , Plastocianina/metabolismo , Espectrofotometria Ultravioleta , Espectroscopia de Luz Próxima ao Infravermelho/instrumentação , Spinacia oleracea/química , Synechocystis/química , Tabaco/química
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