RESUMEN
Human skin is exposed to various physical and chemical stress factors, which commonly cause the oxidation of lipids and proteins. In this study, azo initiator AAPH [2,2' -azobis(2-methylpropionamidine) dihydrochloride] was employed to initiate lipid peroxidation in porcine skin as an ex vivo model for human skin. We demonstrate that malondialdehyde (MDA), a secondary product of lipid peroxidation, is covalently bound to collagen in the dermis, forming MDA-collagen adducts. The binding of MDA to collagen results in an unfolding of the collagen triple helix, formation of the dimer of α-chains of collagen, and fragmentation of the collagen α-chain. It is proposed here that the MDA is bound to the lysine residues of α-chain collagen, which are involved in electrostatic interaction and hydrogen bonding with the glutamate and aspartate of other α-chains of the triple helix. Our data provide crucial information about the MDA binding topology in the skin, which is necessary to understand better the various types of skin-related diseases and the aging process in the skin under stress.
Asunto(s)
Colágeno , Estrés Oxidativo , Humanos , Porcinos , Malondialdehído/metabolismo , Oxidación-Reducción , Colágeno/metabolismo , Peroxidación de Lípido , AnimalesRESUMEN
Photosystem II (PSII) is an intrinsic membrane protein complex that functions as a light-driven water:plastoquinone oxidoreductase in oxygenic photosynthesis. Electron transport in PSII is associated with formation of reactive oxygen species (ROS) responsible for oxidative modifications of PSII proteins. In this study, oxidative modifications of the D1 and D2 proteins by the superoxide anion (O2â¢-) and the hydroxyl (HOâ¢) radicals were studied in WT and a tocopherol cyclase (vte1) mutant, which is deficient in the lipid-soluble antioxidant α-tocopherol. In the absence of this antioxidant, high-resolution tandem mass spectrometry was used to identify oxidation of D1:130E to hydroxyglutamic acid by O2â¢- at the PheoD1 site. Additionally, D1:246Y was modified to either tyrosine hydroperoxide or dihydroxyphenylalanine by O2â¢- and HOâ¢, respectively, in the vicinity of the nonheme iron. We propose that α-tocopherol is localized near PheoD1 and the nonheme iron, with its chromanol head exposed to the lipid-water interface. This helps to prevent oxidative modification of the amino acid's hydrogen that is bonded to PheoD1 and the nonheme iron (via bicarbonate), and thus protects electron transport in PSII from ROS damage.
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Aminoácidos/química , Arabidopsis/enzimología , Complejo de Proteína del Fotosistema II/química , Superóxidos/química , Tilacoides/enzimología , alfa-Tocoferol/química , Aminoácidos/metabolismo , Arabidopsis/genética , Arabidopsis/efectos de la radiación , Sitios de Unión , Radical Hidroxilo/química , Radical Hidroxilo/metabolismo , Transferasas Intramoleculares/química , Transferasas Intramoleculares/genética , Transferasas Intramoleculares/metabolismo , Hierro/química , Hierro/metabolismo , Luz , Modelos Moleculares , Mutación , Oxidación-Reducción , Oxígeno/química , Oxígeno/metabolismo , Fotosíntesis/fisiología , Fotosíntesis/efectos de la radiación , Complejo de Proteína del Fotosistema II/genética , Complejo de Proteína del Fotosistema II/metabolismo , Unión Proteica , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Superóxidos/metabolismo , Termodinámica , Thermosynechococcus/enzimología , Thermosynechococcus/genética , Thermosynechococcus/efectos de la radiación , Tilacoides/genética , Tilacoides/efectos de la radiación , alfa-Tocoferol/metabolismoRESUMEN
Oxidative processes in all types of organisms cause the chemical formation of electronically excited species, with subsequent ultraweak photon emission termed biological auto(chemi)luminescence (BAL). Imaging this luminescence phenomenon using ultrasensitive devices could potentially enable monitoring of oxidative stress in optically accessible areas of the human body, such as skin. Although oxidative stress induced by UV light has been explored, for chemically induced stress, there is no in vivo-quantified imaging of oxidative processes in human skin using BAL under the controlled extent of oxidative stress conditions. Furthermore, the mechanisms and dynamics of BAL from the skin have not been fully explored. Here, we demonstrate that different degrees of chemically induced oxidative stress on the skin can be spatially resolved quantitatively through noninvasive label-free BAL imaging. Additionally, to gain insight into the underlying mechanisms, a minimal chemical model of skin based on a mixture of lipid, melanin, and water was developed and used to show that it can be used to reproduce essential features of the response of real skin to oxidative stress. Our results contribute to novel, noninvasive photonic label-free methods for quantitative sensing of oxidative processes and oxidative stress.
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Luminiscencia , Piel , Humanos , Piel/metabolismo , Estrés Oxidativo , Rayos Ultravioleta , FotonesRESUMEN
Geothermal energy installations are becoming increasingly common in new city developments and renovations. With a broad range of technological applications and improvements in this field, the demand for suitable monitoring technologies and control processes for geothermal energy installations is also growing. This article identifies opportunities for the future development and deployment of IoT sensors applied to geothermal energy installations. The first part of the survey describes the technologies and applications of various sensor types. Sensors that monitor temperature, flow rate and other mechanical parameters are presented with a technological background and their potential applications. The second part of the article surveys Internet-of-Things (IoT), communication technology and cloud solutions applicable to geothermal energy monitoring, with a focus on IoT node designs, data transmission technologies and cloud services. Energy harvesting technologies and edge computing methods are also reviewed. The survey concludes with a discussion of research challenges and an outline of new areas of application for monitoring geothermal installations and innovating technologies to produce IoT sensor solutions.
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Energía Geotérmica , Internet de las Cosas , Nube Computacional , Tecnología de la Información , TecnologíaRESUMEN
Skin plays an important role in protection, metabolism, thermoregulation, sensation, and excretion whilst being consistently exposed to environmental aggression, including biotic and abiotic stresses. During the generation of oxidative stress in the skin, the epidermal and dermal cells are generally regarded as the most affected regions. The participation of reactive oxygen species (ROS) as a result of environmental fluctuations has been experimentally proven by several researchers and is well known to contribute to ultra-weak photon emission via the oxidation of biomolecules (lipids, proteins, and nucleic acids). More recently, ultra-weak photon emission detection techniques have been introduced to investigate the conditions of oxidative stress in various living systems in in vivo, ex vivo and in vitro studies. Research into two-dimensional photon imaging is drawing growing attention because of its application as a non-invasive tool. We monitored spontaneous and stress-induced ultra-weak photon emission under the exogenous application of a Fenton reagent. The results showed a marked difference in the ultra-weak photon emission. Overall, these results suggest that triplet carbonyl (3C=O∗) and singlet oxygen (1O2) are the final emitters. Furthermore, the formation of oxidatively modified protein adducts and protein carbonyl formation upon treatment with hydrogen peroxide (H2O2) were observed using an immunoblotting assay. The results from this study broaden our understanding of the mechanism of the generation of ROS in skin layers and the formation/contribution of various excited species can be used as tools to determine the physiological state of the organism.
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Peróxido de Hidrógeno , Piel , Especies Reactivas de Oxígeno/metabolismo , Peróxido de Hidrógeno/metabolismo , Piel/metabolismo , Estrés Oxidativo , Oxidación-Reducción , Proteínas/metabolismoRESUMEN
The largest stable photosystem II (PSII) supercomplex in land plants (C2S2M2) consists of a core complex dimer (C2), two strongly (S2) and two moderately (M2) bound light-harvesting protein (LHCB) trimers attached to C2 via monomeric antenna proteins LHCB4-6. Recently, we have shown that LHCB3 and LHCB6, presumably essential for land plants, are missing in Norway spruce (Picea abies), which results in a unique structure of its C2S2M2 supercomplex. Here, we performed structure-function characterization of PSII supercomplexes in Arabidopsis (Arabidopsis thaliana) mutants lhcb3, lhcb6, and lhcb3 lhcb6 to examine the possibility of the formation of the "spruce-type" PSII supercomplex in angiosperms. Unlike in spruce, in Arabidopsis both LHCB3 and LHCB6 are necessary for stable binding of the M trimer to PSII core. The "spruce-type" PSII supercomplex was observed with low abundance only in the lhcb3 plants and its formation did not require the presence of LHCB4.3, the only LHCB4-type protein in spruce. Electron microscopy analysis of grana membranes revealed that the majority of PSII in lhcb6 and namely in lhcb3 lhcb6 mutants were arranged into C2S2 semi-crystalline arrays, some of which appeared to structurally restrict plastoquinone diffusion. Mutants without LHCB6 were characterized by fast induction of non-photochemical quenching and, on the contrary to the previous lhcb6 study, by only transient slowdown of electron transport between PSII and PSI. We hypothesize that these functional changes, associated with the arrangement of PSII into C2S2 arrays in thylakoids, may be important for the photoprotection of both PSI and PSII upon abrupt high-light exposure.
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Proteínas de Arabidopsis/genética , Arabidopsis/genética , Proteínas de Unión a Clorofila/genética , Complejo de Proteína del Fotosistema II/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Unión a Clorofila/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Picea/metabolismoRESUMEN
Reactive oxygen species (ROS) are formed in photosystem II (PSII) under various types of abiotic and biotic stresses. It is considered that ROS play a role in chloroplast-to-nucleus retrograde signaling, which changes the nuclear gene expression. However, as ROS lifetime and diffusion are restricted due to the high reactivity towards biomolecules (lipids, pigments, and proteins) and the spatial specificity of signal transduction is low, it is not entirely clear how ROS might transduce signal from the chloroplasts to the nucleus. Biomolecule oxidation was formerly connected solely with damage; nevertheless, the evidence appears that oxidatively modified lipids and pigments are be involved in chloroplast-to-nucleus retrograde signaling due to their long diffusion distance. Moreover, oxidatively modified proteins show high spatial specificity; however, their role in signal transduction from chloroplasts to the nucleus has not been proven yet. The review attempts to summarize and evaluate the evidence for the involvement of ROS in oxidative signaling in PSII.
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Cloroplastos , Complejo de Proteína del Fotosistema II , Cloroplastos/metabolismo , Lípidos , Estrés Oxidativo , Complejo de Proteína del Fotosistema II/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/fisiologíaRESUMEN
The acclimation of higher plants to different light intensities is associated with a reorganization of the photosynthetic apparatus. These modifications, namely, changes in the amount of peripheral antenna (LHCII) of photosystem (PS) II and changes in PSII/PSI stoichiometry, typically lead to an altered chlorophyll (Chl) a/b ratio. However, our previous studies show that in spruce, this ratio is not affected by changes in growth light intensity. The evolutionary loss of PSII antenna proteins LHCB3 and LHCB6 in the Pinaceae family is another indication that the light acclimation strategy in spruce could be different. Here we show that, unlike Arabidopsis, spruce does not modify its PSII/PSI ratio and PSII antenna size to maximize its photosynthetic performance during light acclimation. Its large PSII antenna consists of many weakly bound LHCIIs, which form effective quenching centers, even at relatively low light. This, together with sensitive photosynthetic control on the level of cytochrome b6f complex (protecting PSI), is the crucial photoprotective mechanism in spruce. High-light acclimation of spruce involves the disruption of PSII macro-organization, reduction of the amount of both PSII and PSI core complexes, synthesis of stress proteins that bind released Chls, and formation of "locked-in" quenching centers from uncoupled LHCIIs. Such response has been previously observed in the evergreen angiosperm Monstera deliciosa exposed to high light. We suggest that, in contrast to annuals, shade-tolerant evergreen land plants have their own strategy to cope with light intensity changes and the hallmark of this strategy is a stable Chl a/b ratio.
Asunto(s)
Arabidopsis , Picea , Aclimatación , Arabidopsis/metabolismo , Clorofila/metabolismo , Clorofila A/metabolismo , Complejo de Citocromo b6f/metabolismo , Citocromos b/metabolismo , Proteínas de Choque Térmico/metabolismo , Luz , Complejos de Proteína Captadores de Luz/metabolismo , Complejo de Proteína del Fotosistema I/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Picea/metabolismoRESUMEN
Reactive oxygen species (ROS) represent a group of molecules with a signaling role that are involved in regulating human cell proliferation and differentiation. Increased ROS concentrations are often associated with the local nonspecific oxidation of biological macromolecules, especially proteins and lipids. Free radicals, in general, may randomly damage protein molecules through the formation of protein-centered radicals as intermediates that, in turn, decay into several end oxidation products. Malondialdehyde (MDA), a marker of free-radical-mediated lipid oxidation and cell membrane damage, forms adducts with proteins in a nonspecific manner, leading to the loss of their function. In our study, we utilized U-937 cells as a model system to unveil the effect of four selected bioactive compounds (chlorogenic acid, oleuropein, tomatine, and tyrosol) to reduce oxidative stress associated with adduct formation in differentiating cells. The purity of the compounds under study was confirmed by an HPLC analysis. The cellular integrity and changes in the morphology of differentiated U-937 cells were confirmed with confocal microscopy, and no significant toxicity was found in the presence of bioactive compounds. From the Western blot analysis, a reduction in the MDA adduct formation was observed in cells treated with compounds that underlaid the beneficial effects of the compounds tested.
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Estrés Oxidativo , Radicales Libres/metabolismo , Humanos , Malondialdehído , Oxidación-Reducción , Especies Reactivas de Oxígeno/farmacologíaRESUMEN
Free radical-mediated activation of inflammatory macrophages remains ambiguous with its limitation to study within biological systems. U-937 and HL-60 cell lines serve as a well-defined model system known to differentiate into either macrophages or dendritic cells in response to various chemical stimuli linked with reactive oxygen species (ROS) production. Our present work utilizes phorbol 12-myristate-13-acetate (PMA) as a stimulant, and factors such as concentration and incubation time were considered to achieve optimized differentiation conditions. ROS formation likely hydroxyl radical (HOâ) was confirmed by electron paramagnetic resonance (EPR) spectroscopy combined with confocal laser scanning microscopy (CLSM). In particular, U-937 cells were utilized further to identify proteins undergoing oxidation by ROS using anti-DMPO (5,5-dimethyl-1-pyrroline N-oxide) antibodies. Additionally, the expression pattern of NADPH Oxidase 4 (NOX4) in relation to induction with PMA was monitored to correlate the pattern of ROS generated. Utilizing macrophages as a model system, findings from the present study provide a valuable source for expanding the knowledge of differentiation and protein expression dynamics.
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Diferenciación Celular , Radicales Libres/metabolismo , Monocitos/citología , Monocitos/metabolismo , Proteínas/metabolismo , Acetofenonas/farmacología , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Forma de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Espectroscopía de Resonancia por Spin del Electrón , Células HL-60 , Humanos , Radical Hidroxilo , Monocitos/efectos de los fármacos , NADP/metabolismo , Coloración y Etiquetado , Acetato de Tetradecanoilforbol/farmacología , Células U937RESUMEN
A suite of untargeted methods has been applied for the characterization of aerosol from the Tobacco Heating System 2.2 (THS2.2), a heated tobacco product developed by Philip Morris Products S.A. and commercialized under the brand name IQOS®. A total of 529 chemical constituents, excluding water, glycerin, and nicotine, were present in the mainstream aerosol of THS2.2, generated by following the Health Canada intense smoking regimen, at concentrations ≥ 100 ng/item. The majority were present in the particulate phase (n = 402), representing more than 80% of the total mass determined by untargeted screening; a proportion were present in both particulate and gas-vapor phases (39 compounds). The identities for 80% of all chemical constituents (representing > 96% of the total determined mass) were confirmed by the use of authentic analytical reference materials. Despite the uncertainties that are recognized to be associated with aerosol-based untargeted approaches, the reported data remain indicative that the uncharacterized fraction of TPM generated by THS2.2 has been evaluated to the fullest practicable extent. To the best of our knowledge, this work represents the most comprehensive chemical characterization of a heated tobacco aerosol to date. Graphical abstract.
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Aerosoles/análisis , Productos de Tabaco/análisis , Cromatografía de Gases y Espectrometría de Masas , Calor , Humo/análisis , Nicotiana/químicaRESUMEN
The use of flavoring substances is an important element in the development of reduced-risk products for adult smokers to increase product acceptance and encourage switching from cigarettes. In a first step towards characterizing the sub-chronic inhalation toxicity of neat flavoring substances, a study was conducted using a mixture of the substances in a base solution of e-liquid, where the standard toxicological endpoints of the nebulized aerosols were supplemented with transcriptomics analysis. The flavor mixture was produced by grouping 178 flavors into 26 distinct chemical groups based on structural similarities and potential metabolic and biological effects. Flavoring substances predicted to show the highest toxicological effect from each group were selected as the flavor group representatives (FGR). Following Organization for Economic Cooperation and Development Testing Guideline 413, rats were exposed to three concentrations of the FGR mixture in an e-liquid composed of nicotine (23 µg/L), propylene glycol (1520 µg/L), and vegetable glycerin (1890 µg/L), while non-flavored and no-nicotine mixtures were included as references to identify potential additive or synergistic effects between nicotine and the flavoring substances. The results indicated that the inhalation of an e-liquid containing the mixture of FGRs caused very minimal local and systemic toxic effects. In particular, there were no remarkable clinical (in-life) observations in flavored e-liquid-exposed rats. The biological effects related to exposure to the mixture of neat FGRs were limited and mainly nicotine-mediated, including changes in hematological and blood chemistry parameters and organ weight. These results indicate no significant additive biological changes following inhalation exposure to the nebulized FGR mixture above the nicotine effects measured in this sub-chronic inhalation study. In a subsequent study, e-liquids with FGR mixtures will be aerosolized by thermal treatment and assessed for toxicity.
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Cigarrillo Electrónico a Vapor/toxicidad , Sistemas Electrónicos de Liberación de Nicotina , Aromatizantes/toxicidad , Perfilación de la Expresión Génica , Hígado/efectos de los fármacos , Sistema Respiratorio/efectos de los fármacos , Transcriptoma/efectos de los fármacos , Vapeo/efectos adversos , Animales , Biomarcadores/sangre , Seguridad de Productos para el Consumidor , Femenino , Exposición por Inhalación , Hígado/metabolismo , Hígado/patología , Masculino , Ratas Sprague-Dawley , Sistema Respiratorio/inmunología , Sistema Respiratorio/metabolismo , Sistema Respiratorio/patología , Medición de Riesgo , Factores de Tiempo , Pruebas de ToxicidadRESUMEN
The Photosystem II reaction center is vulnerable to photoinhibition. The D1 and D2 proteins, lying at the core of the photosystem, are susceptible to oxidative modification by reactive oxygen species that are formed by the photosystem during illumination. Using spin probes and EPR spectroscopy, we have determined that both O2â¢- and HO⢠are involved in the photoinhibitory process. Using tandem mass spectroscopy, we have identified a number of oxidatively modified D1 and D2 residues. Our analysis indicates that these oxidative modifications are associated with formation of HO⢠at both the Mn4O5Ca cluster and the nonheme iron. Additionally, O2â¢- appears to be formed by the reduction of O2 at either PheoD1 or QA Early oxidation of D1:332H, which is coordinated with the Mn1 of the Mn4O5Ca cluster, appears to initiate a cascade of oxidative events that lead to the oxidative modification of numerous residues in the C termini of the D1 and D2 proteins on the donor side of the photosystem. Oxidation of D2:244Y, which is a bicarbonate ligand for the nonheme iron, induces the propagation of oxidative reactions in residues of the D-de loop of the D2 protein on the electron acceptor side of the photosystem. Finally, D1:130E and D2:246M are oxidatively modified by O2â¢- formed by the reduction of O2 either by PheoD1â¢- or QAâ¢- The identification of specific amino acid residues oxidized by reactive oxygen species provides insights into the mechanism of damage to the D1 and D2 proteins under light stress.
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Aminoácidos/metabolismo , Oxidación-Reducción , Complejo de Proteína del Fotosistema II/metabolismo , Aminoácidos/química , Antioxidantes/metabolismo , Cloruros/metabolismo , Espectroscopía de Resonancia por Spin del Electrón , Radical Hidroxilo/metabolismo , Espectrometría de Masas , Modelos Moleculares , Conformación Molecular , Oxígeno/metabolismo , Complejo de Proteína del Fotosistema II/química , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Leaf senescence, accompanied by chlorophyll breakdown, chloroplast degradation and inhibition of photosynthesis, can be suppressed by an exogenous application of cytokinins. Two aromatic cytokinin arabinosides (6-benzylamino-9-ß-d-arabinofuranosylpurines; BAPAs), 3-hydroxy- (3OHBAPA) and 3-methoxy- (3MeOBAPA) derivatives, have recently been found to possess high anti-senescence activity. Interestingly, their effect on the maintenance of chlorophyll content and maximal quantum yield of photosystem II (PSII) in detached dark-adapted leaves differed quantitatively in wheat (Triticum aestivum L. cv. Aranka) and Arabidopsis (Arabidopsisthaliana L. (Col-0)). In this work, we have found that the anti-senescence effects of 3OHBAPA and 3MeOBAPA in wheat and Arabidopsis also differ in other parameters, including the maintenance of carotenoid content and chloroplasts, rate of reduction of primary electron acceptor of PSII (QA) as well as electron transport behind QA, and partitioning of absorbed light energy in light-adapted leaves. In wheat, 3OHBAPA had a higher protective effect than 3MeOBAPA, whereas in Arabidopsis, 3MeOBAPA was the more efficient derivative. We have found that the different anti-senescent activity of 3OHBAPA and 3MeOBAPA was coupled to different ethylene production in the treated leaves: the lower the ethylene production, the higher the anti-senescence activity. 3OHBAPA and 3MeOBAPA also efficiently protected the senescing leaves of wheat and Arabidopsis against oxidative damage induced by both H2O2 and high-light treatment, which could also be connected with the low level of ethylene production.
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Arabidopsis/metabolismo , Senescencia Celular , Citocininas/farmacología , Etilenos/metabolismo , Reguladores del Crecimiento de las Plantas/farmacología , Hojas de la Planta/metabolismo , Triticum/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/crecimiento & desarrollo , Fotosíntesis , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/crecimiento & desarrollo , Triticum/efectos de los fármacos , Triticum/crecimiento & desarrolloRESUMEN
Prenylquinols (tocochromanols and plastoquinols) serve as efficient physical and chemical quenchers of singlet oxygen (1 O2 ) formed during high light stress in higher plants. Although quenching of 1 O2 by prenylquinols has been previously studied, direct evidence for chemical quenching of 1 O2 by plastoquinols and their oxidation products is limited in vivo. In the present study, the role of plastoquinol-9 (PQH2 -9) in chemical quenching of 1 O2 was studied in Arabidopsis thaliana lines overexpressing the SOLANESYL DIPHOSPHATE SYNTHASE 1 gene (SPS1oex) involved in PQH2 -9 and plastochromanol-8 biosynthesis. In this work, direct evidence for chemical quenching of 1 O2 by plastoquinols and their oxidation products is presented, which is obtained by microscopic techniques in vivo. Chemical quenching of 1 O2 was associated with consumption of PQH2 -9 and formation of its various oxidized forms. Oxidation of PQH2 -9 by 1 O2 leads to plastoquinone-9 (PQ-9), which is subsequently oxidized to hydroxyplastoquinone-9 [PQ(OH)-9]. We provide here evidence that oxidation of PQ(OH)-9 by 1 O2 results in the formation of trihydroxyplastoquinone-9 [PQ(OH)3 -9]. It is concluded here that PQH2 -9 serves as an efficient 1 O2 chemical quencher in Arabidopsis, and PQ(OH)3 -9 can be considered as a natural product of 1 O2 reaction with PQ(OH)-9. The understanding of the mechanisms underlying 1 O2 chemical quenching provides information on the role of plastoquinols and their oxidation products in the response of plants to photooxidative stress.
RESUMEN
Nontargeted screening methodologies are powerful approaches for comprehensive chemical characterization of complex matrixes. In order to maximize chemical space coverage, three analytical methods using two-dimensional gas chromatography with time-of-flight mass spectrometry for nonpolar, polar, and volatile compounds have been established. The structural identification process was streamlined with an in-house developed computer-assisted structure identification platform, which facilitated the identification of novel compounds and also delivered semiquantitative concentrations for all compounds. Key performance parameters for this nontargeted platform, including chemical space coverage, confidence for structural identification, accuracy of semiquantification, and performance of differential analysis, were evaluated. The automated structural identification process was assessed using a subset of 243 compounds (out of 2990), which were confirmed to be present in cigarette smoke using reference standards. Consistently high true positive identification rates between 88.2% and 96.2% across the different concentration ranges investigated were demonstrated. Accuracy for semiquantification was assessed by comparison with quantitative data from literature, where a maximum 4-fold deviation from available targeted analysis values was estimated.
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Cromatografía de Gases y Espectrometría de Masas/métodos , Contaminación por Humo de Tabaco/análisis , Cromatografía de Gases y Espectrometría de Masas/estadística & datos numéricosRESUMEN
The Arabidopsis vte1 mutant is devoid of tocopherol and plastochromanol (PC-8). When exposed to excess light energy, vte1 produced more singlet oxygen (1 O2 ) and suffered from extensive oxidative damage compared with the wild type. Here, we show that overexpressing the solanesyl diphosphate synthase 1 (SPS1) gene in vte1 induced a marked accumulation of total plastoquinone (PQ-9) and rendered the vte1 SPS1oex plants tolerant to photooxidative stress, indicating that PQ-9 can replace tocopherol and PC-8 in photoprotection. High total PQ-9 levels were associated with a noticeable decrease in 1 O2 production and higher levels of Hydroxyplastoquinone (PQ-C), a 1 O2 -specific PQ-9 oxidation product. The extra PQ-9 molecules in the vte1 SPS1oex plants were stored in the plastoglobules and the chloroplast envelopes, rather than in the thylakoid membranes, whereas PQ-C was found almost exclusively in the thylakoid membranes. Upon exposure of wild-type plants to high light, the thylakoid PQ-9 pool decreased, whereas the extrathylakoid pool remained unchanged. In vte1 and vte1 SPS1oex plants, the PQ-9 losses in high light were strongly amplified, affecting also the extrathylakoid pool, and PQ-C was found in high amounts in the thylakoids. We conclude that the thylakoid PQ-9 pool acts as a 1 O2 scavenger and is replenished from the extrathylakoid stock.
Asunto(s)
Depuradores de Radicales Libres/metabolismo , Plastoquinona/metabolismo , Oxígeno Singlete/metabolismo , Tilacoides/metabolismo , Transferasas Alquil y Aril/metabolismo , Proteínas de Arabidopsis/metabolismo , Clorofila/metabolismo , Cloroplastos/metabolismo , Cromatografía Líquida de Alta Presión , Espectroscopía de Resonancia por Spin del Electrón , Luz , Peroxidación de Lípido , Estrés Oxidativo/efectos de la radiaciónRESUMEN
Light and cytokinins are known to be the key players in the regulation of plant senescence. In detached leaves, the retarding effect of light on senescence is well described; however, it is not clear to what extent is this effect connected with changes in endogenous cytokinin levels. We have performed a detailed analysis of changes in endogenous content of 29 cytokinin forms in detached leaves of Arabidopsis thaliana (wild-type and 3 cytokinin receptor double mutants). Leaves were kept under different light conditions, and changes in cytokinin content were correlated with changes in chlorophyll content, efficiency of photosystem II photochemistry, and lipid peroxidation. In leaves kept in darkness, we have observed decreased content of the most abundant cytokinin free bases and ribosides, but the content of cis-zeatin increased, which indicates the role of this cytokinin in the maintenance of basal leaf viability. Our findings underscore the importance of light conditions on the content of specific cytokinins, especially N6 -(Δ2 -isopentenyl)adenine. On the basis of our results, we present a scheme summarizing the contribution of the main active forms of cytokinins, cytokinin receptors, and light to senescence regulation. We conclude that light can compensate the disrupted cytokinin signalling in detached leaves.
Asunto(s)
Arabidopsis/metabolismo , Citocininas/metabolismo , Hojas de la Planta/metabolismo , Envejecimiento/metabolismo , Envejecimiento/efectos de la radiación , Arabidopsis/efectos de la radiación , Clorofila/metabolismo , Luz , Peroxidación de Lípido , Malondialdehído/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Hojas de la Planta/efectos de la radiaciónRESUMEN
BACKGROUND: Photosystem II proteins of higher plant chloroplasts are prone to oxidative stress, and most prominently the reaction center-binding D1 protein is damaged under abiotic stress. The reactive oxygen species produced under these stress conditions have been suggested to be responsible for the protein injury. SCOPE OF REVIEW: Recently, it has been shown that the primary and secondary products of non-enzymatic and enzymatic lipid peroxidation have a capability to modify photosystem II proteins. Here, we give an overview showing how lipid peroxidation products formed under light stress and heat stress in the thylakoid membranes cause oxidative modification of proteins in higher plant photosystem II. MAJOR CONCLUSIONS: Damage to photosystem II proteins by lipid peroxidation products represents a new mechanism underlying photoinhibition and heat inactivation. GENERAL SIGNIFICANCE: Complete characterization of photosystem II protein damage is of crucial importance because avoidance of the damage makes plants to survive under various abiotic stresses. Further physiological significance of photosystem II protein oxidation by lipid peroxidation product should have a potential relevance to plant acclimation because the oxidized proteins might serve as signaling molecules.