HX600, a synthetic agonist for RXR-Nurr1 heterodimer complex, prevents ischemia-induced neuronal damage.

Ischemic stroke is amongst the leading causes of death and disabilities. The available treatments are suitable for only a fraction of patients and thus novel therapies are urgently needed. Blockage of one of the cerebral arteries leads to massive and persisting inflammatory reaction contributing to the nearby neuronal damage. Targeting the detrimental pathways of neuroinflammation has been suggested to be beneficial in conditions of ischemic stroke. Nuclear receptor 4A-family (NR4A) member Nurr1 has been shown to be a potent modulator of harmful inflammatory reactions, yet the role of Nurr1 in cerebral stroke remains unknown. Here we show for the first time that an agonist for the dimeric transcription factor Nurr1/retinoid X receptor (RXR), HX600, reduces microglia expressed proinflammatory mediators and prevents inflammation induced neuronal death in in vitro co-culture model of neurons and microglia. Importantly, HX600 was protective in a mouse model of permanent middle cerebral artery occlusion and alleviated the stroke induced motor deficits. Along with the anti-inflammatory capacity of HX600 in vitro, treatment of ischemic mice with HX600 reduced ischemia induced Iba-1, p38 and TREM2 immunoreactivities, protected endogenous microglia from ischemia induced death and prevented leukocyte infiltration. These anti-inflammatory functions were associated with reduced levels of brain lysophosphatidylcholines (lysoPCs) and acylcarnitines, metabolites related to proinflammatory events. These data demonstrate that HX600 driven Nurr1 activation is beneficial in ischemic stroke and propose that targeting Nurr1 is a novel candidate for conditions involving neuroinflammatory component.

Accumulation of polycyclic aromatic hydrocarbons by lichen transplants: Comparison with gas-phase passive air samplers.

This study compared the accumulation of 16 polycyclic aromatic hydrocarbons (PAHs) in samples of the lichen Evernia prunastri exposed for 3 months in and around an industrial area of S Italy with that in co-located passive gas-phase air samplers. The results showed a strong linear correlations (R=0.96, P<0.05) between total PAHs in lichens and in passive samplers, clearly indicating that lichen transplants may provide direct quantitative information on the atmospheric load by total PAHs, allowing translation of lichen values into atmospheric concentrations. To the best of our knowledge this is the first study reporting such a correlation with gas-phase passive air samplers.

Effects of heavy metals on ultrastructure and Hsp70 induction in Lemna minor L. exposed to water along the Sarno River, Italy.

The effects of freshwater pollution in the highly contaminated river Sarno (Campania, Southern Italy) have been evaluated using bags containing the aquatic plant Lemna minor (Lemnacee, Arales), in order to determine morpho-physiological modifications as a response to pollutants. The exposition of Lemna bags for 7 days on three different sites along the river path showed alterations in chloroplasts and vacuoles shape and organization. Moreover, some specimens were exposed in vitro at the same heavy metal (HM) concentrations measured in the polluted sites of the river, and compared with data from the bag experiment; to verify the dose and time dependent effects, samples were exposed to HM in vitro at concentrations ranging from 10(-6) to 10(-4)M up to 7 days. Transmission electron microscopy (TEM) observations on in vitro plants confirmed that ultrastructural alterations affected most of plastids and the shape of different subcellular structures, namely vacuoles; in in vitro stressed specimens, Heat Shock Proteins 70 (Hsp70) levels changed, in dependence of changing levels of HM measured in different sites along the river path. Thus L. minor exhibited a possible correlation between the levels of HM pollution and Hsp70 occurrence; interestingly, the data presented showed that copper specifically increased Hsp70 levels at concentrations detected in polluted river waters, whereas cadmium and lead did not; on the other side, the latter represent highly toxic elements when specimens were exposed to higher levels in vitro. The effects of specific elements in vitro are compared to those observed in bags exposed along the river path; thus results are examined in order to propose L. minor as an organism able to be utilized to monitor heavy metals pollution; the possibility of using Hsp70s as specific markers of HM pollution is discussed.

Uptake and acute toxicity of cerium in the lichen Xanthoria parietina.

Environmental cerium (Ce) levels are likely to increase in the near future and monitoring of its biological effects will therefore be necessary. The aim of this study was to test if treatment of the lichen Xanthoria parietina with Ce-containing solutions (0.1mM, 1mM, 10mM and 100mM) causes Ce bioaccumulation (both extra- and intra-cellularly) as well as physiological (sample viability, membrane lipids peroxidation, photosynthetic performance, water-soluble proteins content) and ultrastructural alterations. The results showed that treatment with Ce solutions induces Ce bioaccumulation, both extra-cellularly and intra-cellularly, which in turn causes an acute toxicity, evident as decreased sample viability, marked decrease in the photosynthetic performance and important changes in the ultrastructure.

Temporal trends of element concentrations and ecophysiological parameters in the lichen Pseudevernia furfuracea transplanted in and around an industrial area of S Italy.

This study investigated the temporal trends of element concentrations and ecophysiological parameters (integrity of cell membranes, membrane lipid peroxidation, and sample vitality) in thalli of the lichen Pseudevernia furfuracea transplanted in and around an industrial area of S Italy from May 2010 to May 2011, once every 3 months. Element accumulation at levels indicating that the atmosphere of the study area was polluted by most elements occurred only during the third trimester of exposure, and the ecophysiological parameter best reflecting this trend was sample vitality expressed in terms of activity of the enzyme dehydrogenase. Among the available climatological parameters, the different amount of rainfall during the exposure period may influence the results, especially the number of hours with rainfall >16 mm/day, which is the threshold between light-moderate and moderate-heavy rainfall in the Mediterranean area.

Antimony toxicity in the lichen Xanthoria parietina (L.) Th. Fr.

In this paper we tested if treating the lichen Xanthoria parietina with Sb-containing solutions causes Sb bioaccumulation as well as physiological and ultrastructural changes. Total and intracellular antimony content in Sb-treated samples increased progressively with increasing concentration in the treatment solutions. Incubation of X. parietina thalli with Sb at concentrations as low as 0.1mM caused a decrease in sample viability, measured as intensity of respiratory activity, and damage to cell membranes, expressed in terms of membrane lipid peroxidation, as well as ultrastructural changes such as plasmolysis, impairment of the thylakoid system of the alga and cytoplasmic lipid droplets. The photosynthetic system hardly responded, at least under the tested experimental conditions.

Physiological response of the epiphytic lichen Evernia prunastri (L.) Ach. to ecologically relevant nitrogen concentrations.

This study investigated the physiological response of the epiphytic lichen Evernia prunastri to ecologically relevant concentrations of nitrogen compounds. Lichen samples were sprayed for 4 weeks either with water or 50, 150 and 500 µM NH(4)Cl. The integrity of cell membranes and chlorophyll a fluorescence emission (F(V)/F(M) and PI(ABS)) were analyzed. No membrane damage occurred after the exposure period. F(V)/F(M), a classical fluorescence indicator, decreased during the second week of treatment with 500 µM NH(4)Cl and the third week with 50 and 150 µM NH(4)Cl. PI(ABS), an overall index of the photosynthetic performance, was more sensitive and decreased already during the first week with 500 µM NH(4)Cl and the second week with 150 µM NH(4)Cl. Since E. prunastri has been exposed to ammonium loads corresponding to real environmental conditions, these findings open the way to an effective use of this species as early indicators of environmental nitrogen excess.

Bioacumulation and ultrastructural effects of Cd, Cu, Pb and Zn in the moss Scorpiurum circinatum (Brid.) Fleisch. & Loeske.

This paper tested if culturing the moss Scorpiurum circinatum (Brid.) Fleisch. & Loeske with metal solutions (Cd, Cu, Pb and Zn) for 30 days causes metal bioaccumulation and ultrastructural changes. The results showed that despite the high heavy metal concentrations in treatment solutions, treated samples did not show severe ultrastructural changes and cells were still alive and generally well preserved. Bioaccumulation highlighted that moss cells survived to heavy metal toxicity by immobilizing most toxic ions extracellularly, likely in binding sites of the cell wall, which is the main site of metal detoxification.

Long-term biological monitoring of environmental quality around a solid waste landfill assessed with lichens.

The diversity of epiphytic lichens and the accumulation of selected trace elements in the lichen Flavoparmelia caperata L. (Hale) were used as indicators of pollution around a landfill in central Italy along 14 years of waste management. Lichens revealed an increased deposition for some elements (i.e., Cd, Cr, Fe and Ni) and a decrease of the lichen diversity at sites facing the landfill after an enlargement of the dumping area. However, the results allowed to exclude a significant increase in heavy metal depositions in the surrounding area and suggested that successful waste management may be associated with environmental quality. It is concluded that lichen monitoring might provide essential information to enhance the implementation of ecological impact assessment, supporting industrial regulatory procedures, also when waste management is concerned.

The integrity of lichen cell membrane as a suitable parameter for monitoring biological effects of acute nitrogen pollution.

This study aimed at testing the suitability of cell membrane integrity in the lichen Evernia prunastri (L.) Ach. as sensitive indicator of nitrogen (N) stress, to set up a rapid and effective method for monitoring biological effects of acute N pollution. Lichen samples were incubated in solutions of potassium nitrate, ammonium nitrate and ammonium sulphate at different concentrations, and cell membrane damage, expressed in terms of increased electrolyte leakage, was measured after 0, 24, 48 and 96 h. Cell membrane damage was observed in E. prunastri in the presence of high or very high N concentrations, irrespective of the compound supplied. Since the mycobiont represents the large majority of a lichen biomass, it is reasonable to assume that ion leakage mainly occurred from fungal cells. Although in biomonitoring studies the photobiont is usually regarded as the most sensitive partner of the lichen symbiosis, our findings suggest that the mycobiont is most affected in the case of N-excess, and that this feature can be used as suitable indicator of acute N stress episodes.

Heavy metal deposition in the Italian "triangle of death" determined with the moss Scorpiurum circinatum.

In this study, a biomonitoring project using the moss Scorpiurum circinatum was carried out to evaluate the deposition and biological effects of heavy metals in the area of Acerra (Naples, S Italy), one of the vertices of the sadly called "Italian triangle of death" owing to the dramatic increase in tumours. The results clearly indicated that the study area is heavily polluted by heavy metals, a large proportion of which is likely present in the atmosphere in particulate form. The ultrastructural organization of exposed samples was essentially preserved, but cell membrane pits, cytoplasm vesicles and concentric multilamellar/multivesicular bodies, probably induced by pollution, were found, which may be involved in the tolerance mechanisms to metal pollution in this moss species. Although severe biological effects were not found at the ultrastructural level in the exposed moss, effects on humans, especially after long-term exposure, are to be expected.

Do polyamines alter the sensitivity of lichens to nitrogen stress?

The sensitivity of lichens measuring photosynthetic efficiency and polyamines as modulator of nitrogen stress tolerance was investigated. Two lichen species with a markedly different tolerance to nitrogen compounds, namely Evernia prunastri (L.) Ach. and Xanthoria parietina (L.) Th.Fr., were incubated with deionized water (control) and solutions of KNO(3), NH(4)NO(3) and (NH(4))(2)SO(4) and then exposed to different light conditions. The F(v)/F(m) parameter (maximum quantum efficiency of photosystem II) was used as stress indicator. The results showed that F(v)/F(m) values, in the produced experimental conditions, were independent from the light gradient. Photosynthetic efficiency of E. prunastri was impaired by high ammonium concentrations, while nitrate had no effect; X. parietina was hardly influenced by nitrogen compounds. External supply of polyamines reduced the sensitivity of E. prunastri, while polyamine inhibitors reduced the tolerance of X. parietina to NH(4)(+), suggesting that polyamines play an important role in modulating the sensitivity/tolerance to nitrogen stress.

Integrated biomonitoring of air quality with plants and lichens: a case study on ambient ozone from central Italy.

A biennial integrated survey, based on the use of vascular plants for the bioindication of the effects of tropospheric ozone together with the use of automatic analysers of ozone, as well as the mapping of lichen biodiversity was performed in the area of Castelfiorentino (Tuscany, central Italy). Photochemically produced ozone proved to be a fundamental presence during the warm season, with maximum hourly means reaching 114 ppb, exceeding the information threshold as fixed by EU: the use of supersensitive tobacco Bel-W3 confirmed the opportunity of carrying out detailed cost-effective monitoring surveys. The potential for didactical and educational implications of this methodology are appealing. Critical levels set up for the protection of vegetation have exceeded considerably. The comparison of biomass productivity in sensitive and resistant individuals (NC-S and NC-R white clover clones, in the framework of an European network) provided evidence that ambient ozone levels are associated with relevant reduction (up to 30%) in the performance of sensitive material; effects on flowering were also pronounced. The economic assessment of such an impact deserves attention. Mapping of epiphytic lichen biodiversity--which has been used to monitor air quality worldwide--was not related to ozone geographical distribution as depicted by tobacco response.

Comparison between the accumulation capacity of four lichen species transplanted to a urban site.

The capacity to accumulate trace elements from the atmosphere of the lichens Hypogymnia physodes, Parmelia sulcata, Pseudevernia furfuracea and Usnea gr. hirta transplanted to an urban site of N Italy was compared. Twenty-nine elements (Al, As, Br, Ca, Cd, Ce, Cl, Co, Cr, Cs, Cu, Fe, Hg, I, K, La, Mg, Mn, Ni, Pb, Rb, Sb, Sc, Se, Sm, Th, Ti, V, Zn) were analyzed by Instrumental Neutron Activation Analysis (INAA) and Electro-Thermal Atomic Absorption Spectroscopy (ET-AAS). The ratio between the concentrations of each element in exposed samples to that of control samples (exposed-to-control ratio and EC ratio) was used to investigate the accumulation rates of lichen thalli. The results showed that in general elements did not exhibit well defined trends, but rather showed fluctuations, and indicated that H. physodes, P. furfuracea and U. gr. hirta have a similar accumulation capacity, while that of P. sulcata is lower.

Lichen biomonitoring of ammonia emission and nitrogen deposition around a pig stockfarm.

Effects of high ammonia emissions and nitrogen deposition were investigated on lichens around a pig stockfarm (ca. 7,000 animals) in central Italy. Four sites were selected along a transect at 200, 400, 1000 and 2500 m from the stockfarm, the diversity of epiphytic lichens was measured and transplanted thalli of Xanthoria parietina and Flavoparmelia caperata exposed, together with passive NH3 (diffusion tubes) samplers. Ammonia dramatically decreased from the centre of the stockfarm to the sampled sites, where it was correlated with bark pH. Total lichen diversity was not associated with either NH3 concentrations or bark pH, but the diversity of strictly nitrophytic species was highly correlated with both parameters. Physconia grisea was the best indicator species for NH3 pollution. Total N accumulated in X. parietina and F. caperata was correlated with NH3 concentrations.

Effects of NO2 and NH3 from road traffic on epiphytic lichens.

The results of a survey aimed at investigating whether NO2 and NH3 emitted by road traffic can influence lichen diversity, lichen vitality and the accumulation of nitrogen in lichen thalli are reported. For this purpose, distance from a highway in a rural environment of central Italy was regarded as the main parameter to check this hypothesis. The results of the present survey indicated that road traffic is not a relevant source of NH3. On the other hand, NO2 concentrations, although rather low, were negatively correlated with distance from the highway according to a typical logarithmic function. No association between NO2 concentrations and the diversity of epiphytic lichens was found, probably because of the low NO2 values measured. Also bark properties were not influenced by distance from the highway. Accumulation of nitrogen, reduction in the content of chlorophyll a, chlorophyll b and total carotenoids were found in transplanted thalli of Evernia prunastri, but NO2 was not responsible for these changes, which were probably caused by applications of N-based fertilizers.

Biodiversity of epiphytic lichens and heavy metal contents of Flavoparmelia caperata thalli as indicators of temporal variations of air pollution in the town of Montecatini Terme (central Italy).

The results of a biomonitoring survey carried out in the town of Montecatini Terme (central Italy) in the period 1993-2000 using the biodiversity of epiphytic lichens and the accumulation of heavy metals in thalli of Flavoparmelia caperata as indicators of air pollution are reported. From 1993 to 2000, the mean ILD value for the whole study area increased from 18+/-18 to 42+/-22, the lowest ILD value increased from 0 to 11 and the highest ILD value increased from 67 to 84. A 'lichen desert' was found only in 1993 and stations classified as 'natural' were lacking only in 1993 and 1996. Concentrations of all heavy metals decreased from 1993 to 1999. It is concluded that vehicular traffic is the main source of atmospheric pollution in the study area. The most likely changes that may have caused lichen communities to improve and heavy metals to drop is conversion of heating systems to methane (abatement of SO2) and the use of unleaded gasoline (reduction of Pb). The present results showed that despite their slow growth rate, lichens respond rapidly to decreasing concentrations of air pollutants, allowing annual changes to be detected.

Determination of baseline element composition of lichens using samples from high elevations.

The results of a survey aimed at providing baseline element composition of lichens from unpolluted or very low polluted areas are reported. Lichen samples collected at high elevation areas in Himalayas (Nepal), Mt. Kenya (Kenya) and the Alps (Italy) were analyzed for their trace element content, mostly by INAA, allowing for a multi-element analysis. The elemental composition of lichen samples was essentially influenced by natural occurrence, mainly airborne soil dust. However, also anthropogenic input determined by long-distance atmospheric transport of pollutants was involved for some highly volatile elements such as Br, Cd and Sb. To avoid the influence of soil contamination and air pollution, and obtain baseline concentrations to be used as reference (the "natural" elemental composition of lichens), the average of the lowest concentrations is suggested.

Lichens as biomonitors of uranium in the Balkan area.

The contribution of the conflict of 1999 to the environmental levels of uranium in the Balkan area was evaluated by means of lichens used as biomonitors. The average U concentration found in lichens in the present study was in line with the values reported for lichens from other countries and well below the levels found in lichens collected in areas with natural or anthropogenic sources of U. Measurement of isotopic ratios 235U/238U allowed to exclude the presence of depleted uranium. According to these results, we could not detect widespread environmental contamination by depleted uranium in the Balkan area.

Lichen distribution and bioindicator tobacco plants give discordant response: a case study from Italy.

Epiphytic lichen biodiversity (LB) distribution was evaluated in a 1000 km2 area in Tuscany (Central Italy). In the same area, a survey was performed to monitor tropospheric ozone (O3) phytotoxic effects using tobacco plants (Nicotiana tabacum) cv. Bel-W3 (O3 supersensitive) and cv. Bel-B (O3 resistant) as bioindicators. The LB proved to be negatively correlated with ambient levels of nitrogen oxides, particulate and carbon monoxide, but not with benzene. LB spatial distribution followed a peculiar trend, with highest values in areas with low population density. Data from an O3 analyser set in evidence a typical circadian profile, confirming the photochemical nature of this pollutant. Quite high nocturnal values along the coast were related to the presence of surrounding mountains which constrained the back-and-forth flow of the air in sea breezes. Long- and short-term phytotoxicity critical levels for O3 were systematically trespassed. Bel-W3 tobacco was always affected in every site involved in the study; average O3 injury distribution was greater in rural areas; tobacco response was positively correlated to several O3 descriptors with second-order functions. Correlation analysis failed to demonstrate any association between lichen and tobacco data, due to the fact that LB values were higher in inland zones, far from the main urban and industrial areas, where instead damage to tobacco plants was generally lower. It is concluded that the lichen biodiversity is not suitable for monitoring O3 levels, at least in the study area.