RESUMEN
Leech infestation poses a significant threat to Mithun (Bos frontalis) in the north-eastern region of India, leading to various health issues and potential fatality. To address this pressing concern, we conducted a comprehensive research study aimed at assessing the efficacy of herbal plant extracts against aquatic leeches, particularly Tyrannobdella rex, and land leeches of the Philobdella sp. Our investigation involved the evaluation of six distinct plant extracts, with a focus on their ability to combat leech infestation. The results of our study revealed that among the various plant extracts tested, only the ethanolic extracts of soapnut (Sapindus mukorossi) and tobacco (Nicotiana tabacum) exhibited notable effectiveness in combating aquatic leeches. At a concentration of 5%, these extracts displayed significant lethality, with soapnut extract achieving a remarkable kill time of 6.0 ± 0.40 min, while tobacco extract showed a kill time of 31.5 ± 1.32 min. In the case of land leeches, tobacco extract proved to be highly efficient, with an average kill time of 1.5 ± 0.28 min at a 5% concentration. Soapnut extract also exhibited effectiveness against land leeches, albeit with a slightly longer kill time of 14.25 ± 1.10 min at the same concentration. Additionally, Litsea grass oil (Litsea cubeba) demonstrated promising efficacy against both aquatic and land leeches, suggesting its potential as a versatile leech control agent. These compelling findings have significant implications for the management and control of leech infestation among Mithun populations. By identifying and harnessing the leech-repelling properties of soapnut, tobacco, and Litsea grass oil, this research offers practical and environmentally friendly solutions for mitigating the adverse effects of leech infestation. Furthermore, the insights gained from this study pave the way for the development of innovative strategies to safeguard the health and well-being of Mithun in the future.
Asunto(s)
Sanguijuelas , Extractos Vegetales , Animales , Extractos Vegetales/farmacología , Sanguijuelas/efectos de los fármacos , India , Nicotiana/química , Infestaciones Ectoparasitarias/parasitología , Infestaciones Ectoparasitarias/tratamiento farmacológico , Infestaciones Ectoparasitarias/veterinaria , EtanolRESUMEN
Oral cancer is the most common malignancy in many developing countries, such as India, due to increased consumption of smokeless tobacco. The trace elemental components in commercially packaged forms of tobacco can play a significant role in the pathogenesis of oral cancer. To qualitatively assess the trace elements in various types of commercially packaged forms of tobacco using laser-induced breakdown spectroscopy (LIBS). Two popular varieties of 'Paan masala' that contained a mixture of slaked lime with areca nut, catechu, and other flavouring agents (tobacco was absent) and four types of packaged tobacco were obtained from 'Paan' shops. The contents in the packets were made into pellets using a hydraulic press and subjected to elemental analysis using LIBS. A ten-trial experiment was carried out on all six pellets. The National Institute of Standards and Technology (NIST) database was used to assess the emission lines. The elements obtained from commercially packaged tobacco and Paan masala were similar: calcium (Ca), iron (Fe), aluminium (Al), nickel (Ni), and chromium (Cr). Substances that cause DNA damage and carcinogenesis are inorganic elements such as nickel. Our study revealed that carcinogens such as nickel are present in the commercially packaged forms of tobacco and 'Paan masala' samples.
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Nicotiana , Oligoelementos , Oligoelementos/análisis , Nicotiana/química , Análisis Espectral/métodos , Níquel/análisis , Rayos Láser , Productos de Tabaco/análisis , Embalaje de Productos , Tabaco sin Humo/análisis , Cromo/análisis , Calcio/análisis , Humanos , Hierro/análisisRESUMEN
Although deterioration of silicone maxillofacial prostheses is severely accentuated in smoking patients, the phenomenon has not been systematically studied. To address a gap in the literature concerning the stability of maxillofacial prostheses during service, in this contribution, the effect of cigarette smoke on the aspect and physical properties of M511 silicone elastomer was evaluated. The aspect, surface, and overall properties of the silicone material, pigmented or not, were followed by AFM, color measurements, FTIR, water contact angle measurements, TGA-DTG and DSC, hardness and compression stress-strain measurements. The types of the contaminants adsorbed were assessed by XRF, ESI-MS, MALDI-MS, and NMR spectral analyses. Important modifications in color, contact angle, surface roughness, local mechanical properties, and thermal properties were found in the silicone material for maxillofacial prostheses after exposure to cigarettes smoke. The presence of lead, nicotine, and several other organic compounds adsorbed into the silicone material was emphasized. Slight decrease in hardness and increase in Young's modulus was found. The combined data show important impact of cigarette smoke on the silicone physical properties and could indicate chemical transformations by secondary cross-linking. To our knowledge, this is the first study making use of complementary physical methods to assess the effect of cigarette smoke on the aspect and integrity of silicone materials for maxillofacial prostheses.
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Ensayo de Materiales , Prótesis Maxilofacial , Humo , Humanos , Elastómeros de Silicona/química , Nicotiana/química , ColorRESUMEN
In recent years, nanocarrier-based pesticide delivery systems have provided new possibilities for the efficient utilization of pesticides. In this research, we developed a hydroxypropyl-ß-cyclodextrin-modified graphene oxide (GO-HP-ß-CD) nanocarrier for pyraclostrobin (Pyr) delivery and studied its application for tobacco target spot disease control. GO-HP-ß-CD has excellent pesticide-loading performance for Pyr (adsorption capacity of 1562.5 mg/g) and good water dispersibility and stability. Besides, GO-HP-ß-CD shows pH-responsive release performance. In addition, GO-HP-ß-CD also has better leaf affinity than Pyr, and it can effectively adhere to the leaf surface after simulated washing. The results of antifungal experiments indicate that GO-HP-ß-CD-Pyr has a good preventive effect on tobacco target spot disease, and its EC50 value is 0.384 mg/L, which is lower than Pyr. Specifically, this nanopesticide formulation does not contain toxic organic solvent or additive, so it has good environmental friendliness. Therefore, we believe that the GO-HP-ß-CD-Pyr nanopesticide has brilliant potential in the prevention and control of tobacco diseases.
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Grafito , Nicotiana , Estrobilurinas , Grafito/química , Nicotiana/química , Estrobilurinas/química , Antifúngicos/química , Antifúngicos/farmacología , Enfermedades de las Plantas/prevención & control , Enfermedades de las Plantas/microbiología , Carbamatos/química , Portadores de Fármacos/química , Plaguicidas/química , beta-Ciclodextrinas/química , Fungicidas Industriales/química , Fungicidas Industriales/farmacologíaRESUMEN
Tobacco flavors are extensively utilized in traditional tobacco products, electronic nicotine, heated tobacco products, and snuff. To inhibit fungal growth arising from high moisture content, preservatives such as benzoic acid (BA), sorbic acid (SA), and parabens are often incorporated into tobacco flavors. Nonetheless, consuming preservatives beyond safety thresholds may pose health risks. Therefore, analytical determination of these preservatives is crucial for both quality assurance and consumer protection. For example, BA and SA can induce adverse reactions in susceptible individuals, including asthma, urticaria, metabolic acidosis, and convulsions. Parabens, because of their endocrine activity, are classified as endocrine-disrupting chemicals. Despite extensive research, the concurrent quantification of trace-level hydrophilic (BA and SA) and hydrophobic (methylparaben, ethylparaben, isopropylparaben, propylparaben, butylparaben, isobutylparaben, and benzylparaben) preservatives in tobacco flavors remains challenging. Traditional liquid phase extraction coupled with high performance liquid chromatography (HPLC) often results in high false positive rates and inadequate sensitivity. In contrast, tandem mass spectrometry offers high sensitivity and specificity; however, its widespread application is limited by laborious sample preparation and significant operational costs. Therefore, it is crucial to establish a fast and sensitive sample pretreatment and analysis method for the nine preservatives in tobacco flavors. In this study, a method for the simultaneous determination of the nine preservatives (SA, BA and seven parabens) in tobacco flavor was established based on three phase-hollow fiber-liquid phase microextraction (3P-HF-LPME) technology combined with HPLC. To obtain the optimal pretreatment conditions, extraction solvent type, sample phase pH, acceptor phase pH, sample phase volume, extraction time, and mass fraction of sodium chloride, were examined. Additionally, the HPLC parameters, including UV detection wavelength and mobile phase composition, were refined. The optimal extraction conditions were as follows: dihexyl ether was used as extraction solvent, 15 mL sample solution (pH 4) was used as sample phase, sodium hydroxide aqueous solution (pH 12) was used as acceptor phase, and the extraction was carried out at 800 r/min for 30 min. Chromatographic separation was accomplished using an Agilent Poroshell 120 EC-C18 column (100 mm×3 mm, 2.7 µm) and a mobile phase comprising methanol, 0.02 mol/L ammonium acetate aqueous solution (containing 0.5% acetic acid), and acetonitrile for gradient elution. Under the optimized conditions, the nine target analytes showed good linear relationships in their respective linear ranges, the correlation coefficients (r) were ≥0.9967, limits of detection (LODs) and quantification (LOQs) were 0.02-0.07 mg/kg and 0.08-0.24 mg/kg, respectively. Under two spiked levels, the enrichment factors (EFs) and extraction recoveries (ERs) of the nine target analytes were 30.6-91.1 and 6.1%-18.2%, respectively. The recoveries of the nine target analytes ranged from 82.2% to 115.7% and the relative standard deviations (RSDs) (n=5) were less than 14.5% at low, medium and high levels. The developed method is straightforward, precise, sensitive, and well-suited for the rapid screening of preservatives in tobacco flavor samples.
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Microextracción en Fase Líquida , Parabenos , Conservadores Farmacéuticos , Cromatografía Líquida de Alta Presión , Parabenos/análisis , Microextracción en Fase Líquida/métodos , Conservadores Farmacéuticos/análisis , Ácido Benzoico/análisis , Nicotiana/química , Ácido Sórbico/análisis , Aromatizantes/análisis , Productos de Tabaco/análisisRESUMEN
Tobacco flavor, an important tobacco additive, is an essential raw material in cigarette production that can effectively improve the quality of tobacco products, add aroma and taste, and increase the suction flavor. The quality consistency of tobacco flavors affects the quality stability of branded cigarettes. Therefore, the quality control of tobacco flavors is a major concern for cigarette and flavor manufacturers. Physical and chemical indices, odor similarity, and sensory efficacy are employed to evaluate the quality of tobacco flavors, and the analysis of chemical components in tobacco flavors is usually conducted using gas chromatography (GC) and high performance liquid chromatography (HPLC). However, because the composition of tobacco flavors is complex, their quality cannot be fully reflected using a single component or combination of components. Therefore, establishing an objective analytical method for the quality control of tobacco flavors is of extreme importance. Chromatographic fingerprint analysis is routinely used for the discriminative analysis of tobacco flavors. Chromatographic fingerprints refer to the general characteristics of the concentration profiles of different chemical compounds. In the daily procurement process, fingerprints established by GC and HPLC are effective for the evaluation and identification of tobacco flavors. However, given continuous improvements in aroma-imitation technology, some flavors with high similarity cannot be directly distinguished using existing methods. In this study, a method for the determination of organic acids and inorganic anions in tobacco flavors based on ion chromatography (IC) was developed to ensure the quality consistency of tobacco flavors. A 1.0 g sample of tobacco flavors and 10 mL of deionized water were mixed and vibrated for 30 min. The aqueous sample solution was passed through a 0.45 µm membrane filter and RP pretreatment column in succession to eliminate interferences and then subjected to IC. Standard solutions containing nine organic acids and seven inorganic anions were used to identify the anions in the tobacco flavors, and satisfactory reproducibility was obtained. The relative standard deviations (RSDs) for retention times and peak areas were <0.71% and <6.02%, respectively. The chromatographic fingerprints of four types of tobacco flavors (samples A-D) from five different batches were obtained. Nine tobacco flavor samples from different manufacturers (samples AY1-AY3, BY1-BY2, CY1-CY2, DY1-DY2) were also analyzed to obtain their chromatographic fingerprints. Hierarchical cluster and similarity analyses were used to evaluate the quality of tobacco flavors from different manufacturers. Hierarchical clustering refers to the process of subdividing a group of samples into clusters that exhibit a high degree of intracluster similarity and intercluster dissimilarity. The dendrograms obtained using SPSS 12.0 indicated good quality consistency among the samples in different batches. Samples AY3, BY2, CY2, and DY1 clustered with the batches of standard tobacco flavors. Therefore, hierarchical cluster analysis can effectively distinguish the quality of products from different manufacturers. The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (version 2.0) was used to evaluate the similarity between the standard tobacco flavors and products from different manufacturers. Among the samples analyzed, samples AY3, BY2, CY2, and DY1 showed the highest similarity values (>97.7%), which was consistent with the results of the hierarchical cluster analysis. This finding indicates that IC combined with chromatographic fingerprint analysis could accurately determine the quality of tobacco flavors. GC combined with ultrasonic-assisted liquid-liquid extraction was also used to analyze the tobacco flavors and verify the accuracy of the proposed method. Compared with GC coupled with ultrasonic-assisted liquid-liquid extraction, IC demonstrated more significant quality differences among certain tobacco flavors.
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Nicotiana , Control de Calidad , Nicotiana/química , Aromatizantes/análisis , Productos de Tabaco/análisis , Cromatografía Líquida de Alta Presión/métodos , Cromatografía de Gases/métodos , Cromatografía por Intercambio Iónico/métodosRESUMEN
Mildewed tobacco leaves seriously impact on cigarette product quality and pose a health risk to person. However, early moldy tobacco leaves are hardly found by naked eyes in the workshop. In this work, we self-assemble AuAg nanoalloys on silicon wafers to construct Si/AuAg chips. The headspace-surface enhanced Raman scattering (SERS) protocol is developed to monitor volatile 1,2-dichloro-3-methoxybenzene (2,3-DCA) and 2,4,6-trichloroanisole (2,4,6-TCA) released from postharvest tobacco. Consequently, the visualization of the SERS peak at 1592 cm-1 assigned to ν(CC) after headspace collection for 10 min and the SERS intensity ratio of 1054 and 1035 cm-1 from 2,3-DCA and 2,4,6-TCA less than 0.5 could be used as indicators to predict early moldy tobacco. Additionally, with headspace collection time prolonging to 2 h, a SERS band at 682 cm-1 due to ν(CCl) of 2,4,6-TCA occurs, confirming the mildew of leaves. The headspace-SERS protocol paves a path for rapid and on-site inspection of the quality of tobacco leaves and cigarettes during storage with a portable Raman system.
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Oro , Nicotiana , Hojas de la Planta , Plata , Espectrometría Raman , Hojas de la Planta/química , Nicotiana/química , Espectrometría Raman/métodos , Plata/química , Oro/química , Anisoles/análisis , Anisoles/química , Nanopartículas del Metal/química , Silicio/química , Enfermedades de las Plantas/microbiologíaRESUMEN
A set of nine unique tobacco extract samples was analyzed using a self-developed electronic nose (E-nose) system, a commercial E-nose, and gas chromatography-mass spectrometry (GC-MS). The evaluation employed principal component analysis, statistical quality control, and soft independent modeling of class analogies (SIMCA). These multifaceted statistical methods scrutinized the collected data. Subsequently, a quality control model was devised to assess the stability of the sample quality. The results showed that the custom E-nose system could successfully distinguish between tobacco extracts with similar odors. After further training and the development of a quality control model for accepted tobacco extracts, it was possible to identify samples with normal and abnormal quality. To further validate our E-nose and extend its use within the tobacco industry, we collected and accurately classified the flavors of different tobacco leaf positions, with a remarkable accuracy rate of 0.9744. This finding facilitates the practical application of our E-nose system for the efficient identification of tobacco leaf positions.
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Nariz Electrónica , Cromatografía de Gases y Espectrometría de Masas , Nicotiana , Hojas de la Planta , Nicotiana/química , Hojas de la Planta/química , Cromatografía de Gases y Espectrometría de Masas/métodos , Odorantes/análisis , Análisis de Componente Principal , Control de Calidad , Aromatizantes/análisisRESUMEN
Chronic obstructive pulmonary disease (COPD), the third leading cause of death worldwide, is caused by chronic exposure to toxic particles and gases, such as cigarette smoke. Free radicals, which are produced during a stress response to toxic particles, play a crucial role in disease progression. Measuring these radicals is difficult since the complex mixture of chemicals within cigarette smoke interferes with radical detection. We used a new quantum sensing technique called relaxometry to measure free radicals with nanoscale resolution on cells from COPD patients and healthy controls exposed to cigarette smoke extract (CSE) or control medium. Epithelial cells from COPD patients display a higher free radical load than those from healthy donors and are more vulnerable to CSE. We show that epithelial cells of COPD patients are more susceptible to the damaging effects of cigarette smoke, leading to increased release of free radicals.
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Bronquios , Células Epiteliales , Enfermedad Pulmonar Obstructiva Crónica , Humo , Humanos , Radicales Libres , Células Epiteliales/efectos de los fármacos , Células Epiteliales/patología , Células Epiteliales/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/patología , Humo/efectos adversos , Bronquios/citología , Bronquios/efectos de los fármacos , Nicotiana/química , Células Cultivadas , Fumar/efectos adversos , Productos de Tabaco/análisis , Productos de Tabaco/efectos adversosRESUMEN
The hydrogen isotopic composition (δ2H) of plant compounds is increasingly used as a hydroclimatic proxy; however, the interpretation of δ2H values is hampered by potential coeffecting biochemical and biophysical processes. Here, we studied δ2H values of water and carbohydrates in leaves and roots, and of leaf n-alkanes, in two distinct tobacco (Nicotiana sylvestris) experiments. Large differences in plant performance and biochemistry resulted from (a) soil fertilization with varying nitrogen (N) species ratios and (b) knockout-induced starch deficiency. We observed a strong 2H-enrichment in sugars and starch with a decreasing performance induced by increasing NO3-/NH4+ ratios and starch deficiency, as well as from leaves to roots. However, δ2H values of cellulose and n-alkanes were less affected. We show that relative concentrations of sugars and starch, interlinked with leaf gas exchange, shape δ2H values of carbohydrates. We thus provide insights into drivers of hydrogen isotopic composition of plant compounds and into the mechanistic modeling of plant cellulose δ2H values.
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Carbohidratos , Hidrógeno , Hojas de la Planta , Hojas de la Planta/química , Hojas de la Planta/metabolismo , Hidrógeno/análisis , Carbohidratos/química , Carbohidratos/análisis , Almidón/química , Nicotiana/química , Lípidos/análisis , Lípidos/química , Raíces de Plantas/química , Raíces de Plantas/metabolismo , Metabolismo de los Hidratos de Carbono , Deuterio/química , Alcanos/análisis , Alcanos/química , Agua/químicaRESUMEN
Neuroinflammation is suggested as one of the potential links between CS-induced neuronal dysfunction. Cigarette smoke (CS) is one of the significant contributors of neuroinflammation, consequently leading to cognitive impairment and neurodegeneration. Microglia are the key resident macrophage cells in the brain with cell surface TLR4 receptor for responding to various stress signals. The CS constituents promote inflammation and oxidative stress in microglia leading to cytotoxicity through the TLR4-MK2 axis. However, the role of MK2 kinase in CS-induced microglial inflammation is not yet clearly understood. Therefore, we have used an MK2 inhibitor, PF-3644022 to study modulation of CS-extract induced oxidative and inflammatory signaling in a mouse microglial cell line, Furthermore, we also evaluated the enzymatic activity of acetylcholinesterase (AChE) on a direct exposure of enzyme with CS. CS exposure led to microglial cytotoxicity and enhanced the level of oxidative stress and proinflammatory cytokine release by microglial cells. The microglial cells pretreated with MK2 inhibitor, PF-3644022 significantly reduced the levels of oxidative stress markers, proinflammatory markers, and improved the level of antioxidant proteins in these cells. In addition, direct exposure of CS showed reduction in the enzymatic activity of AChE.
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Acetilcolinesterasa , Microglía , Estrés Oxidativo , Proteínas Serina-Treonina Quinasas , Animales , Microglía/metabolismo , Microglía/efectos de los fármacos , Ratones , Estrés Oxidativo/efectos de los fármacos , Línea Celular , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Acetilcolinesterasa/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Péptidos y Proteínas de Señalización Intracelular/antagonistas & inhibidores , Humo/efectos adversos , Citocinas/metabolismo , Supervivencia Celular/efectos de los fármacos , Nicotiana/químicaRESUMEN
Chronic obstructive pulmonary disease (COPD) is largely attributed to tobacco smoke exposure. Investigating how airway epithelial cells functionally adapt to tobacco smoke is crucial for understanding the pathogenesis of COPD. The present study was to set up an in vitro model using primary murine airway epithelial cells to mimic the real-life impact of tobacco smoke. Unlike established cell lines, primary cells retain more in vivo-like properties, including growth patterns, aging, and differentiation. These cells exhibit a sensitive inflammatory response and efficient differentiation, thus closely representing physiological conditions. In this model, primary murine airway epithelial cells were cultured for 28 days under an air-liquid interface with an optimal concentration of cigarette smoke extract (CSE), which led to the transformation of a monolayer of undifferentiated cells into a pseudostratified columnar epithelium, indicative of CSE acclimation. Comprehensive multi-omics analyses were then applied to elucidate the mechanisms by which CSE influences the differentiation of basal airway cells. These insights provide a deeper understanding of the cellular processes underpinning COPD progression in response to tobacco smoke exposure.
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Diferenciación Celular , Células Epiteliales , Humo , Animales , Ratones , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Células Epiteliales/citología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Humo/efectos adversos , Nicotiana/química , Mucosa Respiratoria/citología , Mucosa Respiratoria/efectos de los fármacos , Mucosa Respiratoria/metabolismo , Productos de Tabaco , Enfermedad Pulmonar Obstructiva Crónica/patología , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , MultiómicaRESUMEN
OBJECTIVES: To explore the effects and mechanisms of bilirubin on mitochondrial function and type of macrophage cell death after exposure to cigarette smoke extract (CSE). METHODS: RAW264.7 macrophages were treated with different concentrations of CSE and bilirubin solutions and divided into four groups: control, CSE, bilirubin, and bilirubin + CSE groups. The necrotic and apoptotic states of the macrophages were determined using an Annexin V-fluorescein 5-isothiocyanate/propidium iodide (FITC/PI) staining kit. Cytoplasmic NOD-like receptor family, pyrin domain containing 3 (NLRP3) expression in macrophages was detected by immunofluorescence and the levels of IL-1ß and IL-18 in the supernatants of culture medium were detected by enzyme linked immunosorbent assay (ELISA) test. A JC-1 mitochondrial membrane potential detection kit was used to assess mitochondrial membrane damage and the adenosine triphosphate (ATP) assay kit was used to determine intracellular ATP levels. After the macrophages were stained with reactive oxygen species (ROS) specific dye, 2',7'-Dichlorodihydrofluorescein diacetate (DCFH-DA), the fluorescence intensity and proportion of ROS-positive macrophages were measured using flow cytometry. RESULTS: We observed that compared with those of 0 µM (control group), concentrations of 5, 10, or 20 µΜ bilirubin significantly decreased cell viability, which was increased by bilirubin exposure below 1 µM. The effect of CSE on macrophage viability was concentration- and time-dependent. Bilirubin of 0.2 µM could alleviate the inhibition of macrophage viability caused by 5% CSE. In addition, bilirubin intervention could reduce the occurrence of necrosis and pyroptosis to a certain extent. CONCLUSIONS: CSE could cause mitochondrial dysfunction in macrophages, as demonstrated by a decrease in mitochondrial membrane potential and intracellular ATP levels and an increase in ROS production, while bilirubin could relieve mitochondrial dysfunction caused by CSE.
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Bilirrubina , Macrófagos , Mitocondrias , Especies Reactivas de Oxígeno , Animales , Ratones , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Células RAW 264.7 , Especies Reactivas de Oxígeno/metabolismo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Nicotiana/efectos adversos , Nicotiana/química , Humo/efectos adversos , Apoptosis/efectos de los fármacos , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismoRESUMEN
Cigarette smoking is the acknowledged major cause of cancers of the lung and oral cavity and is an established important risk factor for multiple other cancers. DNA addition products (DNA adducts) caused by cigarette smoking are critical factors in its mechanism of carcinogenesis. However, most DNA adducts detected to date in humans cannot be specifically ascribed to smoking but rather have multiple exogenous and endogenous sources. In the study reported here, we prepared [13C]-labeled tobacco to address this problem. We report for the first time the successful growth from seeds to flowering under hydroponic conditions of highly [13C]-labeled tobacco in a controlled 13CO2 environment. The standard growth procedure with optimized conditions is described in detail. The [13C]-enrichment rate was assessed by quantifying nicotine and sugars and their [13C]-isotopologues in this tobacco using high-resolution mass spectrometry, reaching >94% in the tobacco leaves. The [13C]-labeled leaves after curing will be used to make cigarettes, allowing investigation of the specific contributions of tobacco smoke carcinogens to identified DNA adducts in smokers.
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Isótopos de Carbono , Daño del ADN , Hidroponía , Nicotiana , Nicotiana/química , Nicotiana/crecimiento & desarrollo , Nicotiana/metabolismo , Isótopos de Carbono/análisis , Isótopos de Carbono/metabolismo , Humanos , Aductos de ADN/metabolismo , Aductos de ADN/análisis , Fumadores , Hojas de la Planta/química , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/metabolismo , Nicotina/metabolismoRESUMEN
Protoporphyrinogen oxidase (PPO, EC 1.3.3.4) has a high status in the development of new inhibitors. To develop novel and highly effective PPO inhibitors, active substructure linking and bioisosterism replacement strategies were used to design and synthesize novel tetrahydrophthalimide derivatives containing oxadiazole/thiadiazole moieties, and their inhibitory effects on Nicotiana tobacco PPO (NtPPO) and herbicidal activity were evaluated. Among them, compounds B11 (Ki = 9.05 nM) and B20 (Ki = 10.23 nM) showed significantly better inhibitory activity against NtPPO than that against flumiclorac-pentyl (Ki = 46.02 nM). Meanwhile, compounds A20 and B20 were 100% effective against three weeds (Abutilon theophrasti, Amaranthus retroflexus, and Portulaca oleracea) at 37.5 g a.i./ha. It was worth observing that compound B11 was more than 90% effective against three weeds (Abutilon theophrasti, Amaranthus retroflexus, and Portulaca oleracea) at 18.75 and 9.375 g a.i./ha. It was also safer to rice, maize, and wheat than flumiclorac-pentyl at 150 g a.i./ha. In addition, the molecular docking results showed that compound B11 could stably bind to NtPPO and it had a stronger hydrogen bond with Arg98 (2.9 Å) than that of flumiclorac-pentyl (3.2 Å). This research suggests that compound B11 could be used as a new PPO inhibitor, and it could help control weeds in agricultural production.
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Amaranthus , Diseño de Fármacos , Inhibidores Enzimáticos , Herbicidas , Simulación del Acoplamiento Molecular , Oxadiazoles , Ftalimidas , Malezas , Protoporfirinógeno-Oxidasa , Tiadiazoles , Herbicidas/química , Herbicidas/farmacología , Herbicidas/síntesis química , Tiadiazoles/química , Tiadiazoles/farmacología , Tiadiazoles/síntesis química , Malezas/efectos de los fármacos , Malezas/enzimología , Oxadiazoles/química , Oxadiazoles/farmacología , Oxadiazoles/síntesis química , Relación Estructura-Actividad , Ftalimidas/química , Ftalimidas/farmacología , Ftalimidas/síntesis química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Inhibidores Enzimáticos/síntesis química , Protoporfirinógeno-Oxidasa/antagonistas & inhibidores , Protoporfirinógeno-Oxidasa/química , Protoporfirinógeno-Oxidasa/metabolismo , Amaranthus/química , Amaranthus/efectos de los fármacos , Proteínas de Plantas/química , Proteínas de Plantas/antagonistas & inhibidores , Estructura Molecular , Nicotiana/químicaRESUMEN
Reconstituted tobacco (RT) is a product made by reprocessing tobacco waste, experiencing a growing demand for heat-not-burn products. The purpose of this study is to analyze the main flavor ingredients in RT aerosol, as well as the transfer behavior of key flavor substances from substrates to aerosol and the concentrations of these compounds in the substrate after heating. First, we demonstrated that the odor of four RT aerosol samples could be distinguished using an electronic nose. Through non-targeted analysis, 93 volatile compounds were detected by gas chromatography-mass spectrometry, and 286 non/semi-volatile compounds were identified by ultra-high-performance liquid electrophoresis chromatography-mass spectrometry in aerosol. Furthermore, we found that the formation of RT aerosol involves primarily evaporation and distillation, however, the total content delivered from unheated RT samples to aerosol remains relatively low due to compound volatility and cigarette filtration. Thermal reactions during heating indicated the pyrolysis of chlorogenic acid to generate catechol and resorcinol, while Maillard reactions involving glucose and proline produced 2,3-dihydro-3,5-dihydroxy-6-methyl-4h-pyran-4-one. The study highlighted that heating RT at approximately 300°C could mitigate the production of harmful substances while still providing a familiar sensory experience with combusted tobacco.
Asunto(s)
Aromatizantes , Cromatografía de Gases y Espectrometría de Masas , Nicotiana , Aromatizantes/análisis , Aromatizantes/química , Nicotiana/química , Calor , Aerosoles/química , Aerosoles/análisis , Compuestos Orgánicos Volátiles/análisis , Compuestos Orgánicos Volátiles/química , Productos de Tabaco/análisis , Calefacción , Odorantes/análisisRESUMEN
Plants are a newly developing eukaryotic expression system being explored to produce therapeutic proteins. Purification of recombinant proteins from plants is one of the most critical steps in the production process. Typically, proteins were purified from total soluble proteins (TSP), and the presence of miscellaneous intracellular proteins and cytochromes poses challenges for subsequent protein purification steps. Moreover, most therapeutic proteins like antigens and antibodies are secreted to obtain proper glycosylation, and the presence of incompletely modified proteins leads to inconsistent antigen or antibody structures. This work introduces a more effective method to obtain highly purified recombinant proteins from the plant apoplastic space. The recombinant Green fluorescent protein (GFP) is engineered to be secreted into the apoplast of Nicotiana benthamiana and is then extracted using an infiltration-centrifugation method. The GFP-His from the extracted apoplast is then purified by nickel affinity chromatography. In contrast to the traditional methods from TSP, purification from the apoplast produces highly purified recombinant proteins. This represents an important technological improvement for plant production systems.
Asunto(s)
Cromatografía de Afinidad , Proteínas Fluorescentes Verdes , Nicotiana , Nicotiana/genética , Nicotiana/química , Nicotiana/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/química , Proteínas Fluorescentes Verdes/biosíntesis , Cromatografía de Afinidad/métodos , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Centrifugación/métodos , Proteínas Recombinantes de Fusión/aislamiento & purificación , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/biosíntesisRESUMEN
Smoking is a well-established risk factor for several oral diseases, including oral cancer, oral leukoplakia and periodontitis, primarily related to reactive oxygen species (ROS). SS-31, a mitochondria-targeting tetrapeptide, has exhibited demonstrable efficacy in medical conditions by attenuating mitochondrial ROS production. However, its potential in the treatment of oral diseases remains underexplored. The aim of this study was to investigate the therapeutic potential of SS-31 in mitigating smoking-induced oral epithelial injury. Through in vitro experiments, our results indicate that SS-31 plays a protective role against cigarette smoke extract (CSE) by reducing oxidative stress, attenuating inflammatory response, and restoring mitochondrial function. Furthermore, we found that mitophagy, regulated by PINK1 (PTEN-induced putative kinase 1)/Parkin (Parkin RBR E3 ubiquitin-protein ligase), was critical for the protective role of SS-31. Our findings offer valuable insights into SS-31's therapeutic potential in mitigating CSE-induced oxidative stress, inflammatory response, and mitochondrial dysfunction in oral epithelial cells. This study provides novel intervention targets for smoking-related oral diseases.
Asunto(s)
Células Epiteliales , Mitocondrias , Mitofagia , Oligopéptidos , Estrés Oxidativo , Proteínas Quinasas , Humo , Ubiquitina-Proteína Ligasas , Estrés Oxidativo/efectos de los fármacos , Mitofagia/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Humanos , Proteínas Quinasas/metabolismo , Células Epiteliales/metabolismo , Células Epiteliales/efectos de los fármacos , Oligopéptidos/farmacología , Ubiquitina-Proteína Ligasas/metabolismo , Humo/efectos adversos , Especies Reactivas de Oxígeno/metabolismo , Línea Celular , Nicotiana/química , Nicotiana/efectos adversosRESUMEN
Nitroxyl (HNO), a reactive nitrogen species (RNS), is essential for plant growth. However, the action of HNO in plants has been difficult to understand due to the lack of highly sensitive and real-time in-situ monitoring tools. Herein, we presented a near-infrared fluorescent probe, DCI-HNO, based on dicyanoisophorone fluorophore, for real-time mapping HNO in plants. The introduction of a phosphine moiety as a specific HNO recognition unit can inhibit the intramolecular charge transfer (ICT) of probe DCI-HNO. However, in the presence of HNO, the ICT process occurred, leading to the emission at 665 nm. Probe DCI-HNO exhibited high sensitivity (97 nM), rapid response time (8 min), large Stokes shift (135 nm) for detection of HNO in plants. The novel developed probe has successfully imaged endogenous HNO produced during NO/H2S cross-talk in plant tissues. Additionally, the up-regulated in HNO levels during tobacco aging and in response to stress has been confirmed. Therefore, probe DCI-HNO has provided a reliable method for monitoring the NO/H2S cross-talk and revealing the role of HNO in plants.