Potential of PM-selected components to induce oxidative stress and root system alteration in a plant model organism

Over the last years, various acellular assays have been used for the evaluation of the oxidative potential (OP) of particular matter (PM) to predict PM capacity to generate reactive oxygen (ROS) and nitrogen (RNS) species in biological systems. However, relationships among OP and PM toxicological effects on living organisms are still largely unknown. This study aims to assess the effects of atmospheric PM-selected components (brake dust - BD, pellet ash - PA, road dust - RD, certified urban dust NIST1648a - NIST, soil dust - S, coke dust - C and Saharan dust - SD) on the model plant A. thaliana development, with emphasis on their capacity to induce oxidative stress and root morphology alteration. Before growing A. thaliana in the presence of the PM-selected components, each atmospheric dust has been chemically characterized and tested for the OP through dithiothreitol (DTT), ascorbic acid (AA) and 2′,7′-dichlorofluorescin (DCFH) assays. After the exposure, element bioaccumulation in the A. thaliana seedlings, i.e., in roots and shoots, was determined and both morphological and oxidative stress analyses were performed in roots. The results indicated that, except for SD and S, all the tested dusts affected A. thaliana root system morphology, with the strongest effects in the presence of the highest OPs dusts (BD, PA and NIST). Principal component analysis (PCA) revealed correlations among OPs of the dusts, element bioaccumulation and root morphology alteration, identifying the most responsible dust-associated elements affecting the plant. Lastly, histochemical analyses of NO and O2•− content and distribution confirmed that BD, PA and NIST induce oxidative stress in A. thaliana, reflecting the high OPs of these dusts and ultimately leading to cell membrane lipid peroxidation

In-vivo exposure of a plant model organism for the assessment of the ability of PM samples to induce oxidative stress

This study aims to propose an innovative, simple, rapid, and cost-effective method to study oxidative stress induced by PM through in-vivo exposure of the plant model organism Arabidopsis thaliana. A. thaliana seedlings were exposed to urban dust certified for its elemental content and to PM2.5 samples collected in an urban-industrial area of Northern Italy. An innovative technique for the detachment and suspension in water of the whole intact dust from membrane filters was applied to expose the model organism to both the soluble and insoluble fractions of PM2.5, which were analyzed for 34 elements by ICP-MS. Oxidative stress induced by PM on A. thaliana was assessed by light microscopic localization and UV–Vis spectrophotometric determination of superoxide anion (O2−) content on the exposed seedlings by using the nitro blue tetrazole (NBT) assay. The results showed a good efficiency and sensitivity of the method for PM mass concentrations >20 μg m−3 and an increase in O2− content in all exposed seedlings, which mainly depends on the concentration, chemical composition, and sources of the PM administered to the model organism. Particles released by biomass burning appeared to contribute more to the overall toxicity of PM. This method was found to be cost-effective and easy to apply to PM collected on membrane filters in intensive monitoring campaigns in order to obtain valuable information on the ability of PM to generate oxidative stress in living organisms

Nitric oxide cooperates with auxin to mitigate the alterations in the root system caused by cadmium and arsenic

Oryza sativa L. is a worldwide food-crop frequently growing in cadmium (Cd)/arsenic (As) polluted soils, with its root-system as the first target of the pollutants. Root-system development involves the establishment of optimal indole-3-acetic acid (IAA) levels, also requiring the conversion of the IAA natural precursor indole-3-butyric acid (IBA) into IAA, causing nitric oxide (NO) formation. Nitric oxide is a stress-signaling molecule. In rice, a negative interaction of Cd or As with endogenous auxin has been demonstrated, as some NO protective effects. However, a synergism between the natural auxins (IAA and/or IBA) and NO was not yet determined and might be important for ameliorating rice metal(oid)-tolerance. With this aim, the stress caused by Cd/As toxicity in the root cells and the possible recovery by either NO or auxins (IAA/IBA) were evaluated after Cd or As (arsenate) exposure, combined or not with the NO-donor compound sodium-nitroprusside (SNP). Root fresh weight, membrane electrolyte leakage, and H2O2 production were also measured. Moreover, endogenous IAA/IBA contents, transcription-levels of OsYUCCA1 and OsASA2 IAA-biosynthetic-genes, and expression of the IAA-influx-carrier OsAUX1 and the IAA-responsive DR5::GUS construct were analyzed, and NO-epifluorescence levels were measured. Results showed that membrane injury by enhanced electrolyte leakage occurred under both pollutants and was reduced by the treatment with SNP only in Cd-presence. By contrast, no membrane injury was caused by either exogenous NO or IAA or IBA. Cd- and As-toxicity also resulted into a decreased root fresh weight, mitigated by the combination of each pollutant with either IAA or IBA. Cd and As decreased the endogenous NO-content, increased H2O2 formation, and altered auxin biosynthesis, levels and distribution in both adventitious (ARs) and mainly lateral roots (LRs). The SNP-formed NO counteracted the pollutants’ effects on auxin distribution/levels, reduced H2O2 formation in Cd-presence, and enhanced AUX1-expression, mainly in As-presence. Each exogenous auxin, but mainly IBA, combined with Cd or As at 10 µM, mitigated the pollutants’ effects by increasing LR-production and by increasing NO-content in the case of Cd. Altogether, results demonstrate that NO and auxin(s) work together in the rice root system to counteract the specific toxic-effects of each pollutant

Jasmonates, ethylene and brassinosteroids control adventitious and lateral rooting as stress avoidance responses to heavy metals and metalloids

Developmental and environmental signaling networks often converge during plant growth in response to changing conditions. Stress-induced hormones, such as jasmonates (JAs), can influence growth by crosstalk with other signals like brassinosteroids (BRs) and ethylene (ET). Nevertheless, it is unclear how avoidance of an abiotic stress triggers local changes in development as a response. It is known that stress hormones like JAs/ET and BRs can regulate the division rate of cells from the first asymmetric cell divisions (ACDs) in meristems, suggesting that stem cell activation may take part in developmental changes as a stress-avoidance-induced response. The root system is a prime responder to stress conditions in soil. Together with the primary root and lateral roots (LRs), adventitious roots (ARs) are necessary for survival in numerous plant species. AR and LR formation is affected by soil pollution, causing substantial root architecture changes by either depressing or enhancing rooting as a stress avoidance/survival response. Here, a detailed overview of the crosstalk between JAs, ET, BRs, and the stress mediator nitric oxide (NO) in auxin-induced AR and LR formation, with/without cadmium and arsenic, is presented. Interactions essential in achieving a balance between growth and adaptation to Cd and As soil pollution to ensure survival are reviewed here in the model species Arabidopsis and rice

Aldo-keto reductases protect metastatic melanoma from ER stress-independent ferroptosis

The incidence of melanoma is increasing over the years with a still poor prognosis and the lack of a cure able to guarantee an adequate survival of patients. Although the new immuno-based coupled to target therapeutic strategy is encouraging, the appearance of targeted/cross-resistance and/or side effects such as autoimmune disorders could limit its clinical use. Alternative therapeutic strategies are therefore urgently needed to efficiently kill melanoma cells. Ferroptosis induction and execution were evaluated in metastasis-derived wild-type and oncogenic BRAF melanoma cells, and the process responsible for the resistance has been dissected at molecular level. Although efficiently induced in all cells, in an oncogenic BRAF- and ER stress-independent way, most cells were resistant to ferroptosis execution. At molecular level we found that: resistant cells efficiently activate NRF2 which in turn upregulates the early ferroptotic marker CHAC1, in an ER stress-independent manner, and the aldo-keto reductases AKR1C1\u2009\uf7\u20093 which degrades the 12/15-LOX-generated lipid peroxides thus resulting in ferroptotic cell death resistance. However, inhibiting AKRs activity/expression completely resensitizes resistant melanoma cells to ferroptosis execution. Finally, we found that the ferroptotic susceptibility associated with the differentiation of melanoma cells cannot be applied to metastatic-derived cells, due to the EMT-associated gene expression reprogramming process. However, we identified SCL7A11 as a valuable marker to predict the susceptibility of metastatic melanoma cells to ferroptosis. Our results identify the use of pro-ferroptotic drugs coupled to AKRs inhibitors as a new valuable strategy to efficiently kill human skin melanoma cells

Brassinosteroids interact with nitric oxide in the response of rice root systems to arsenic stress

Brassinosteroids (BRs), an emerging class of phytohormones, affect numerous plant physiological and metabolic processes and can improve plant defense systems to counteract metalloid phytotoxicity. Nitric oxide (NO), a reactive nitrogen species (RNS), behaves as a signalling molecule activating plant cellular responses to various environmental conditions. Brassinosteroids induce NO synthesis through nitrate reductase (NR) and NO synthase (NOS) activities. Arsenite and arsenate, inorganic forms of the metalloid arsenic (As), cause both soil pollution and many disorders in numerous plants, including important crops like rice, due to the oxidative stress generated by the imbalance between RNS and reactive oxygen species (ROS). Rice is very susceptible to As toxicity because both As availability and solubility are high in flooded paddy fields in many cultivated areas. The research aims to investigate the effects of BRs on the rice root systems exposed to 10-4 M Na2HAsO40.7 H2O [As(V)] or 2.5 × 10-5 M NaAsO2 [As(III)], highlighting the induced cyto-histological events and dissecting the NO role in the root response. A specific concentration (10-7 M) of 24-epibrassinolide (24-eBL), an exogenously applied BR, increases lateral root (LR) formation of more than 50% in the presence of As(III) or As(V). In addition, eBL attenuates the thickening of the cell walls induced by As in the outermost root cortical layers of LRs and in the adventitious roots (ARs) by reducing of ⁓ 50% the lignin deposition, while it restores the As(v)-altered NO levels by increasing OsNOS1 expression and the cellular NO distribution

Jasmonate promotes auxin-induced adventitious rooting in dark-grown Arabidopsis thaliana seedlings and stem thin cell layers by a cross-talk with ethylene signalling and a modulation of xylogenesis

Background: Adventitious roots (ARs) are often necessary for plant survival, and essential for successful micropropagation. In Arabidopsis thaliana dark-grown seedlings AR-formation occurs from the hypocotyl and is enhanced by application of indole-3-butyric acid (IBA) combined with kinetin (Kin). The same IBA + Kin-treatment induces AR-formation in thin cell layers (TCLs). Auxin is the main inducer of AR-formation and xylogenesis in numerous species and experimental systems. Xylogenesis is competitive to AR-formation in Arabidopsis hypocotyls and TCLs. Jasmonates (JAs) negatively affect AR-formation in de-etiolated Arabidopsis seedlings, but positively affect both AR-formation and xylogenesis in tobacco dark-grown IBA + Kin TCLs. In Arabidopsis the interplay between JAs and auxin in AR-formation vs xylogenesis needs investigation. In de-etiolated Arabidopsis seedlings, the Auxin Response Factors ARF6 and ARF8 positively regulate AR-formation and ARF17 negatively affects the process, but their role in xylogenesis is unknown. The cross-talk between auxin and ethylene (ET) is also important for AR-formation and xylogenesis, occurring through EIN3/EIL1 signalling pathway. EIN3/EIL1 is the direct link for JA and ET-signalling. The research investigated JA role on AR-formation and xylogenesis in Arabidopsis dark-grown seedlings and TCLs, and the relationship with ET and auxin. The JA-donor methyl-jasmonate (MeJA), and/or the ET precursor 1-aminocyclopropane-1-carboxylic acid were applied, and the response of mutants in JA-synthesis and -signalling, and ET-signalling investigated. Endogenous levels of auxin, JA and JA-related compounds, and ARF6, ARF8 and ARF17 expression were monitored. Results: MeJA, at 0.01 μM, enhances AR-formation, when combined with IBA + Kin, and the response of the early-JA-biosynthesis mutant dde2–2 and the JA-signalling mutant coi1–16 confirmed this result. JA levels early change during TCL-culture, and JA/JA-Ile is immunolocalized in AR-tips and xylogenic cells. The high AR-response of the late JA-biosynthesis mutant opr3 suggests a positive action also of 12-oxophytodienoic acid on AR-formation. The crosstalk between JA and ET-signalling by EIN3/EIL1 is critical for AR-formation, and involves a competitive modulation of xylogenesis. Xylogenesis is enhanced by a MeJA concentration repressing AR-formation, and is positively related to ARF17 expression. Conclusions: The JA concentration-dependent role on AR-formation and xylogenesis, and the interaction with ET opens the way to applications in the micropropagation of recalcitrant species

Planck intermediate results. XLI. A map of lensing-induced B-modes

The secondary cosmic microwave background (CMB) $B$-modes stem from the post-decoupling distortion of the polarization $E$-modes due to the gravitational lensing effect of large-scale structures. These lensing-induced $B$-modes constitute both a valuable probe of the dark matter distribution and an important contaminant for the extraction of the primary CMB $B$-modes from inflation. Planck provides accurate nearly all-sky measurements of both the polarization $E$-modes and the integrated mass distribution via the reconstruction of the CMB lensing potential. By combining these two data products, we have produced an all-sky template map of the lensing-induced $B$-modes using a real-space algorithm that minimizes the impact of sky masks. The cross-correlation of this template with an observed (primordial and secondary) $B$-mode map can be used to measure the lensing $B$-mode power spectrum at multipoles up to $2000$. In particular, when cross-correlating with the $B$-mode contribution directly derived from the Planck polarization maps, we obtain lensing-induced $B$-mode power spectrum measurement at a significance level of $12\,\sigma$, which agrees with the theoretical expectation derived from the Planck best-fit $\Lambda$CDM model. This unique nearly all-sky secondary $B$-mode template, which includes the lensing-induced information from intermediate to small ($10\lesssim \ell\lesssim 1000$) angular scales, is delivered as part of the Planck 2015 public data release. It will be particularly useful for experiments searching for primordial $B$-modes, such as BICEP2/Keck Array or LiteBIRD, since it will enable an estimate to be made of the lensing-induced contribution to the measured total CMB $B$-modes.Comment: 20 pages, 12 figures; Accepted for publication in A&A; The B-mode map is part of the PR2-2015 Cosmology Products; available as Lensing Products in the Planck Legacy Archive http://pla.esac.esa.int/pla/#cosmology; and described in the 'Explanatory Supplement' https://wiki.cosmos.esa.int/planckpla2015/index.php/Specially_processed_maps#2015_Lensing-induced_B-mode_ma

Planck intermediate results. XXIX. All-sky dust modelling with Planck, IRAS, and WISE observations

We present all-sky modelling of the high resolution Planck, IRAS, and WISE infrared (IR) observations using the physical dust model presented by Draine and Li in 2007 (DL). We study the performance and results of this model, and discuss implications for future dust modelling. The present work extends the DL dust modelling carried out on nearby galaxies using Herschel and Spitzer data to Galactic dust emission. We employ the DL dust model to generate maps of the dust mass surface density, the optical extinction Av, and the starlight intensity parametrized by Umin. The DL model reproduces the observed spectral energy distribution (SED) satisfactorily over most of the sky, with small deviations in the inner Galactic disk and in low ecliptic latitude areas. We compare the DL optical extinction Av for the diffuse interstellar medium with optical estimates for 2 10^5 quasi-stellar objects (QSOs) observed in the Sloan digital sky survey. The DL Av estimates are larger than those determined towards QSOs by a factor of about 2, which depends on Umin. The DL fitting parameter Umin, effectively determined by the wavelength where the SED peaks, appears to trace variations in the far-IR opacity of the dust grains per unit Av, and not only in the starlight intensity. To circumvent the model deficiency, we propose an empirical renormalization of the DL Av estimate, dependent of Umin, which compensates for the systematic differences found with QSO observations. This renormalization also brings into agreement the DL Av estimates with those derived for molecular clouds from the near-IR colours of stars in the 2 micron all sky survey. The DL model and the QSOs data are used to compress the spectral information in the Planck and IRAS observations for the diffuse ISM to a family of 20 SEDs normalized per Av, parameterized by Umin, which may be used to test and empirically calibrate dust models.Comment: Final version that has appeared in A&

Planck Intermediate Results. IV. The XMM-Newton validation programme for new Planck galaxy clusters

We present the final results from the XMM-Newton validation follow-up of new Planck galaxy cluster candidates. We observed 15 new candidates, detected with signal-to-noise ratios between 4.0 and 6.1 in the 15.5-month nominal Planck survey. The candidates were selected using ancillary data flags derived from the ROSAT All Sky Survey (RASS) and Digitized Sky Survey all-sky maps, with the aim of pushing into the low SZ flux, high-z regime and testing RASS flags as indicators of candidate reliability. 14 new clusters were detected by XMM, including 2 double systems. Redshifts lie in the range 0.2 to 0.9, with 6 clusters at z>0.5. Estimated M500 range from 2.5 10^14 to 8 10^14 Msun. We discuss our results in the context of the full XMM validation programme, in which 51 new clusters have been detected. This includes 4 double and 2 triple systems, some of which are chance projections on the sky of clusters at different z. We find that association with a RASS-BSC source is a robust indicator of the reliability of a candidate, whereas association with a FSC source does not guarantee that the SZ candidate is a bona fide cluster. Nevertheless, most Planck clusters appear in RASS maps, with a significance greater than 2 sigma being a good indication that the candidate is a real cluster. The full sample gives a Planck sensitivity threshold of Y500 ~ 4 10^-4 arcmin^2, with indication for Malmquist bias in the YX-Y500 relation below this level. The corresponding mass threshold depends on z. Systems with M500 > 5 10^14 Msun at z > 0.5 are easily detectable with Planck. The newly-detected clusters follow the YX-Y500 relation derived from X-ray selected samples. Compared to X-ray selected clusters, the new SZ clusters have a lower X-ray luminosity on average for their mass. There is no indication of departure from standard self-similar evolution in the X-ray versus SZ scaling properties. (abridged)Comment: accepted by A&