Urban vegetation and particle air pollution: Experimental campaigns in a traffic hotspot.

This work presents the main results of two experimental campaigns carried out in summer and winter seasons in a complex pollution hotspot near a large park, El Retiro, in Madrid (Spain). These campaigns were aimed at understanding the microscale spatio-temporal variation of ambient concentration levels in areas with high pollution values to obtain data to validate models on the effect of urban trees on particulate matter concentrations. Two different measuring approaches have been used. The first one was static, with instruments continuously characterizing the meteorological variables and the particulate matter concentration outside and inside the park. During the summer campaign, the particulate matter concentration was clearly influenced by a Saharan dust outbreak during the period 23 June to 10 July 2016, when most of the particulate matter was in the fraction PM2.5-10. During the winter campaign, the mass concentrations were related to the meteorological conditions and the high atmospheric stability. The second approach was a dynamic case with mobile measurements by portable instruments. During the summer campaign, a DustTrak instrument was used to measure PM10 and PM2.5 in different transects close to and inside the park at different distances from the traffic lane. It was observed a decrease in the concentrations up to 25% at 20 m and 50% at 200 m. High PM10 values were linked to dust resuspension caused by recreational activities and to a Saharan dust outbreak. The highest PM values were measured at the Independencia square, an area with many bus stops and high traffic density. During the winter campaign, three microaethalometers were used for Black Carbon measurement. Both pollutants also showed a reduction in their concentrations when moving towards inside the park. For PM10 and PM2.5, reductions up to 50% were observed, while for BC this reduction was smaller, about 20%.

Genes coding for transporters showed a rapid and sharp increase in their expression in response to lead, in the aquatic fern (Salvinia minima Baker).

Salvinia minima was assessed for its ability to accumulate lead (Pb) by exposing it to concentrations of 40µM Pb(NO3)2 during 24h. At the same time, the expression levels were quantified, of four genes coding for transporters: SmABCC (ABCC-MRP), SmATPase (ATPase-P3A), SmNhaD (Type-Na+/H+) and SmABCG (ABCG-WBC). In the absence of lead, S. minima had very low expression of those genes, when plants were exposed to the metal however, those genes showed a rapid (in just three hours or less) and sharp increase (up to 60 times) in their expression, particularly the SmNhaD (Type-Na+/H+) gene. This sharp increase in expression levels of the genes studied, occurred at the same time that the plant accumulated the highest content of lead in its tissues. The first two genes, are apparently implicated in detoxification and lead accumulation mechanisms, while the other two genes are apparently involved in maintaining cell balance (homeostatic control) and membrane integrity. Our results confirmed that S. minima is efficient for phytoremediation of water bodies contaminated by lead, as it is efficient in accumulating this metal in its tissues (bioconcentration factor; BCF) values greater than 1000, in short times of exposure. More importantly, our data on the expression profiles of four genes coding for transporters, represent a first sight scenario of the molecular basis for understanding the different mechanism of detoxification, apparently present in this aquatic fern.

Eco-physiological response of Hypnum cupressiforme Hedw. to increased atmospheric ammonia concentrations in a forest agrosystem.

Ammonia (NH3) emissions are linked to eutrophication, plant toxicity and ecosystem shifts from N to P limitation. Bryophytes are key components of terrestrial ecosystems, yet highly sensitive to N deposition. Hence, physiological responses of mosses may be indicative of NH3-related impacts, and thus useful to foresee future ecosystem damages and establish atmospheric Critical Levels (CLEs). In this work, samples of Hypnum cupressiforme Hedw. were seasonally collected along a well-defined NH3 concentration gradient in an oak woodland during a one-year period. We performed a comprehensive evaluation of tissue chemistry, stoichiometry, metabolic enzymes, antioxidant response, membrane damages, photosynthetic pigments, soluble protein content and N and C isotopic fractionation. Our results showed that all the physiological parameters studied (except P, K, Ca and C) responded to the NH3 gradient in predictable ways, although the magnitude and significance of the response were dependent on the sampling season, especially for enzymatic activities and pigments content. Nutritional imbalances, membrane damages and disturbance of cellular C and N metabolism were found as a consequence to NH3 exposure, being more affected the mosses more exposed to the barn atmosphere. These findings suggested significant implications of intensive farming for the correct functioning of oak woodlands and highlighted the importance of seasonal dynamics in the study of key physiological processes related to photosynthesis, mosses nutrition and responses to oxidative stress. Finally, tissue N showed the greatest potential for the identification of NH3-related ecological end points (estimated CLE=3.5µgm-3), whereas highly scattered physiological responses, although highly sensitive, were not suitable to that end.

Identification of up-regulated genes from the metal-hyperaccumulator aquatic fern Salvinia minima Baker, in response to lead exposure.

Lead (Pb) is one of the most serious environmental pollutants. The aquatic fern Salvinia minima Baker is capable to hyper-accumulate Pb in their tissues. However, the molecular mechanisms involved in its Pb accumulation and tolerance capacity are not fully understood. In order to investigate the molecular mechanisms that are activated by S. minima in response to Pb, we constructed a suppression subtractive hybridization library (SSH) in response to an exposure to 40µM of Pb(NO3)2 for 12h. 365 lead-related differentially expressed sequences tags (ESTs) were isolated and sequenced. Among these ESTs, 143 unique cDNA (97 were registered at the GenBank and 46 ESTs were not registered, because they did not meet the GenBank conditions). Those ESTs were identified and classified into 3 groups according to Blast2GO. In terms of metabolic pathways, they were grouped into 29 KEGG pathways. Among the ESTs, we identified some that might be part of the mechanism that this fern may have to deal with this metal, including abiotic-stress-related transcription factors, some that might be involved in tolerance mechanisms such as ROS scavenging, membrane protection, and those of cell homeostasis recovery. To validate the SSH library, 4 genes were randomly selected from the library and analyzed by qRT-PCR. These 4 genes were transcriptionally up-regulated in response to lead in at least one of the two tested tissues (roots and leaves). The present library is one of the few genomics approaches to study the response to metal stress in an aquatic fern, representing novel molecular information and tools to understand the molecular physiology of its Pb tolerance and hyperaccumulation capacity. Further research is required to elucidate the functions of the lead-induced genes that remain classified as unknown, to perhaps reveal novel molecular mechanisms of Pb tolerance and accumulation capacity in aquatic plants.

Prognostic value of metabolic tumour volume and total lesion glycolysis in 18F-FDG PET/CT scans in locally advanced breast cancer staging. / Papel pronóstico del volumen metabólico tumoral y de la glucólisis tumoral total en los estudios 18F-FDG PET/TC de estadificación del cáncer localmente avanzado de mama.

OBJECTIVE: To determine whether metabolic tumour volume (MTV) and total lesion glycolysis (TLG) are able to predict recurrence risk in locally advanced breast cancer (LABC) patients. MATERIAL AND METHODS: Retrospective study of LABC patients who undertook neoadjuvant, local and adjuvant treatment and follow up. A 18F-FDG PET/CT study for initial staging was performed analysing in this study different metabolic parameters (MTV, TLG, SUVmax and SUVmed) both in the primary tumour (T) as well as in axillary nodes (N) and whole-body (WB). RESULTS: Forty females were included between January 2010-2011; follow up until January 2015 was completed. The average follow-up was 46 months. Twenty percent presented recurrence: local disease (n=2) and distant metastasis (n=6); 3 patients died (38% of the patients which recurred and 7.5% from the total). SUVmax, MTV and TLG, in T, N and WB, were higher in those patients with recurrence. The MTV and TLG parameters in the tumour (T) were related to the recurrence rate (P=.020 and P=.028, respectively); whereas SUVmax in the lymph nodes (N) was significantly related (P=.008) to the recurrence rate. The best cut-off points to predict recurrence where: MTV T ≥19.3cm3, TLG T≥74.4g and SUVmax N≥13.8, being 10-12 times more likely to recidivate when these thresholds where exceeded. Tumour grade was the only clinical-pathological variable which was related to recurrence probability (p=.035). CONCLUSIONS: In this study of LABC patients the metabolic parameters which have a better relationship with recurrence rate are: MTV and TLG in the primary tumour, SUVmax in the regional lymph node disease and whole-body PET data.

Throughfall and bulk deposition of dissolved organic nitrogen to holm oak forests in the Iberian Peninsula: Flux estimation and identification of potential sources.

Deposition of dissolved organic nitrogen (DON) in both bulk precipitation (BD) and canopy throughfall (TF) has been measured for the first time in the western Mediterranean. The study was carried out over a year from 2012 to 2013 at four evergreen holm oak forests located in the Iberian Peninsula: two sites in the Province of Barcelona (Northeastern Spain), one in the Province of Madrid (central Spain) and the fourth in the Province of Navarra (Northern Spain). In BD the annual volume weighted mean (VWM) concentration of DON ranged from 0.25 mg l(-1) in Madrid to 1.14 mg l(-1) in Navarra, whereas in TF it ranged from 0.93 mg l(-1) in Barcelona to 1.98 mg l(-1) in Madrid. The contribution of DON to total nitrogen deposition varied from 34% to 56% in BD in Barcelona and Navarra respectively, and from 38% in Barcelona to 72% in Madrid in TF. Agricultural activities and pollutants generated in metropolitan areas were identified as potential anthropogenic sources of DON at the study sites. Moreover, canopy uptake of DON in Navarra was found in spring and autumn, showing that organic nitrogen may be a supplementary nutrient for Mediterranean forests, assuming that a portion of the nitrogen taken up is assimilated during biologically active periods.

Superparamagnetic iron oxide as a tracer for sentinel node biopsy in breast cancer: A comparative non-inferiority study.

AIMS: The gold standard for detection of Sentinel Lymph Nodes (SLN) is a combined radioisotope and blue dye breast injection, using a gamma probe (GP). A new, non-radioactive method was developed, using a tracer (Sienna+(®)) of superparamagnetic iron oxide (SPIO) nanoparticles and a manual magnetometer (SentiMag(®)) (SM). The IMAGINE study was designed to show the non-inferiority of SM compared to GP, for the detection of SLN in breast cancer patients with SLN biopsy indication. METHODS: From November 2013 to June 2014, 181 patients were recruited, and 321 nodes were excised and assessed ex-vivo. Readings from both SM and GP devices were recorded during transcutaneous, intraoperative, and ex-vivo detection attempts. RESULTS: At the patient level, ex-vivo detection rates (primary variable) with SM and GP were 97.8% and 98.3% (concordance rate 99.4%). Transcutaneous and intraoperative detection rates were 95.5% vs 97.2%, and 97.2% vs 97.8% for SM and GP respectively (concordance rates > 97%). At the node level, intraoperative and ex-vivo detection rates were 92.5% vs 89.3% and 91.0% vs 86.3% for SM and GP respectively. In all cases the non-inferiority of SM compared to SM was shown by ruling out a predefined non-inferiority margin of 5%. CONCLUSIONS: Our study showed the non-inferiority of SM as compared to GP. Moreover, the ex-vivo and intraoperative detection rates at the node level were slightly higher with SM.

[Axillary pathologic response after neoadjuvant chemotherapy in locally advanced breast cancer with axillary involvement]. / Respuesta patológica en la axila tras quimioterapia neoadyuvante en el cáncer de mama localmente avanzado con afectación axilar.

AIM: To compare axillary involvement (N+) at initial staging in locally advanced breast cancer (LABC) with axillary lymphadenectomy histologic results after neoadjuvant chemotherapy treatment (NeoChemo). MATERIAL AND METHODS: Retrospective study between November 2011 and September 2013 of LABC cases treated with neoadjuvant chemotherapy based on docetaxel (associated with trastuzumab in HER2 positive cases and carboplatin/adriamycin in HER2 negative cases). Those clinically or radiologically suspected cases of axillary involvement were histologically confirmed. When there was no suspicion of axillary involvement, sentinel lymph node radioguided biopsy (SLNRB) was performed using intradermal injection of (99m)Tc-nanocolloid albumin prior to neoadjuvant treatment. Axillary lymphadenectomy after NeoChemo was undertaken in all cases with positive axilla. Final pathologic response was classified as complete (pCR) when there was no evidence of tumoral disease and as non-pathologic complete response (no pCR) in the opposite case. RESULTS: A total of 346 patients treated with docetaxel were reviewed, identifying 105 LABC. Axillary involvement at initial staging was detected in 70 (67%) before starting NeoChemo. From these 70, 73% (n=51) were N+ (fine needle biopsy and/or biopsy) and the remaining 19 (27%) were occult N+ detected by SLNRB. Axillary lymphadenectomy detected pCR in 56% (39/70), increasing up to 84% pCR when initial N+ status was reached using SNLB. On the other hand, when N+ was detected using fine needle biopsy/lymph biopsy, pCR was only 45%. CONCLUSION: More than 50% of women affected by locally advanced breast cancer with tumoral axillary involvement at initial diagnosis present free metastatic axilla after therapeutic neoadjuvant chemotherapy effect. This increases up to almost 90% in case of occult metastatic axilla detected with sentinel node biopsy prior starting neoadjuvant chemotherapy.

Impact of organ donation in Spanish families: phenomenological approach through relatives' lived experiences.

INTRODUCTION AND PURPOSE: This study explores the family experience when they are told that their recently deceased relative is in a potential position for organ donation, as well as the connotations that develop from that situation and how those nuances have lasted since then for family members. In Spain, and especially in Valencian region (where this study took place), one of the most productive counties for organ donation nationwide, the family interview is considered to be one of the most important tools that transplant coordinators can use to improve and succeed through the process of a potential organ donation. METHODOLOGY AND DATA ANALYSIS: This is a qualitative study based on a phenomenological approach using Giorgi's method for data analysis. Members of 11 families who donated their deceased relatives' organs in La Fe Hospital, Valencia, Spain, from 2009 to 2011, participated through in-depth interviews after signing informed consent agreements. DISCUSSION: Experiences described have shown how the relatives' overview has been affected by the situation in a way that is very different from the professional staff's view of reality. The difference in perspectives creates a barrier which can make the professional team less effective in understanding family interests to create a helpful and successful relationship for organ donation. Understanding the experiences of the families involved can help healthcare providers to conduct a more supportive family interview when the topic is organ donation.

Heavy metal and nitrogen concentrations in mosses are declining across Europe whilst some "hotspots" remain in 2010.

In recent decades, naturally growing mosses have been used successfully as biomonitors of atmospheric deposition of heavy metals and nitrogen. Since 1990, the European moss survey has been repeated at five-yearly intervals. In 2010, the lowest concentrations of metals and nitrogen in mosses were generally found in northern Europe, whereas the highest concentrations were observed in (south-)eastern Europe for metals and the central belt for nitrogen. Averaged across Europe, since 1990, the median concentration in mosses has declined the most for lead (77%), followed by vanadium (55%), cadmium (51%), chromium (43%), zinc (34%), nickel (33%), iron (27%), arsenic (21%, since 1995), mercury (14%, since 1995) and copper (11%). Between 2005 and 2010, the decline ranged from 6% for copper to 36% for lead; for nitrogen the decline was 5%. Despite the Europe-wide decline, no changes or increases have been observed between 2005 and 2010 in some (regions of) countries.

Biomonitoring of traffic-related nitrogen pollution using Letharia vulpina (L.) Hue in the Sierra Nevada, California.

To assess the impact of vehicular emissions on a mixed conifer forest, we measured the contents of the trace elements, N, C, and their respective natural isotopes (δ(15)N and δ(13)C), in the epiphytic lichen, L. vulpina. The samples were collected along transects perpendicular to Interstate 80 (I-80) and along a more remote, secondary forest road (R07). Distance to the road verge, trunk cover, and stand basal area were also recorded. Percent N ranged from 1.10% to 2.00% near I-80 and from 0.78% to 1.13% along R07. Concentrations of N, (15)N, Na, As, Pb, and Zn were enhanced in lichen samples near I-80 and were negatively correlated with distance from the road. Trunk cover values differed between roads (p<0.001) and were negatively correlated with %N (r(2)=0.74; p<0.001). The results indicate that vehicular N emissions are significant enough to alter the surrounding ecosystem, modifying the presence of a sensitive component such as L. vulpina, and suggest that a clean-site threshold of 1.0%N may be too high as an indicator of critical N load exceedance. The study also underscored the potential role of wolf lichen in a large-scale assessment of N deposition and source identification.

Biosynthesis of lead nanoparticles by the aquatic water fern, Salvinia minima Baker, when exposed to high lead concentration.

Salvinia minima Baker is a small floating aquatic fern that is efficient for the removal and storage of heavy metals such as lead and cadmium. In this study, we report that lead removal by S. minima causes large accumulation of lead inside the cells in the form of nanoparticles (PbNPs). The accumulation pattern of lead was analyzed in both, submerged root-like modified fronds (here named "roots"), and in its aerial leaf-like fronds ("leaves"). Analysis by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM) and high resolution transmission electron microscopy (HRTEM) confirmed the biosynthesis of PbNPs by the plant. In both, roots and leaves, PbNPs were found to accumulate almost exclusively at the cell wall and closely associated to the cell membrane. Two types of PbNPs shapes were found in cells of both tissues, those associated to the cell wall were quasi-spherical with 17.2±4.2 nm of diameter, while those associated to the cell membrane/cytoplasm were elongated. Elongated particles were 53.7±29.6 nm in length and 11.1±2.4 nm wide. Infrared spectroscopy (IR) results indicate that cellulose, lignin and pectin are the major components that may be acting as the reducing agents for lead ions; these findings strongly suggest the potential use of this fern to further explore the bio-assisted synthesis of heavy metal nanostructures.

Spatial distribution of PAH concentrations and stable isotope signatures (δ13C, δ15N) in mosses from three European areas--characterization by multivariate analysis.

Polycyclic aromatic hydrocarbon (PAH) concentrations and N, C stable isotope signatures were determined in mosses Hypnum cupressiforme Hedw. from 61 sites of 3 European regions: Île-de-France (France); Navarra (Spain); the Swiss Plateau and Basel area (Switzerland). Total PAH concentrations of 100-700 ng g(-1), as well as δ(13)C values of -32 to -29‰ and δ(15)N values of -11 to -3‰ were measured. Pearson correlation tests revealed opposite trends between high molecular weight PAH (4-6 aromatic rings) content and δ(13)C values. Partial Least Square regressions explained the very significant correlations (r > 0.91, p < 0.001) between high molecular weight PAH concentrations by local urban land use (<10 km) and environmental factors such as elevation and pluviometry. Finally, specific correlations between heavy metal and PAH concentrations were attributed to industrial emissions in Switzerland and road traffic emissions in Spain.

Country-specific correlations across Europe between modelled atmospheric cadmium and lead deposition and concentrations in mosses.

Previous analyses at the European scale have shown that cadmium and lead concentrations in mosses are primarily determined by the total deposition of these metals. Further analyses in the current study show that Spearman rank correlations between the concentration in mosses and the deposition modelled by the European Monitoring and Evaluation Programme (EMEP) are country and metal-specific. Significant positive correlations were found for about two thirds or more of the participating countries in 1990, 1995, 2000 and 2005 (except for Cd in 1990). Correlations were often not significant and sometimes negative in countries where mosses were only sampled in a relatively small number of EMEP grids. Correlations frequently improved when only data for EMEP grids with at least three moss sampling sites per grid were included. It was concluded that spatial patterns and temporal trends agree reasonably well between lead and cadmium concentrations in mosses and modelled atmospheric deposition.

Contribution of the portable gamma camera to detect the sentinel node in breast cancer during surgery.

AIM: To show our experience using the portable gamma camera during surgery in the detection of the sentinel node in breast cancer. MATERIAL AND METHODS: A total of 46 women diagnosed with breast cancer were included. This study was carried out by means of the periareolar intradermal injection. Planar images and SPECT/CT were acquired. On the following day, the sentinel nodes were localized intraoperatively with a portable gamma camera and a gamma probe. Images of the anterior axillary line projection and lateral axillary line projection were acquired before and after extraction of all the sentinel nodes. RESULTS: The images acquired with the portable gamma-camera during the operation showed no activity in all of the patients after the removal of the sentinel nodes. In four out of the 46 cases, the portable gamma camera provided relevant information during the procedure (it detected a sentinel node in two cases in which it had not been detected prior to the surgery and it oriented the surgeon in the localization of low activity nodes when the probe screening had been negative). CONCLUSIONS: The use of the intraoperative portable gamma camera to detect the sentinel node in breast cancer patients is useful in order to ensure, independently of the surgeon, that all the sentinel nodes have been removed. It can also help guide the surgeon in the event of a single low activity sentinel node.

Nitrogen concentrations in mosses indicate the spatial distribution of atmospheric nitrogen deposition in Europe.

In 2005/6, nearly 3000 moss samples from (semi-)natural location across 16 European countries were collected for nitrogen analysis. The lowest total nitrogen concentrations in mosses (<0.8%) were observed in northern Finland and northern UK. The highest concentrations (≥ 1.6%) were found in parts of Belgium, France, Germany, Slovakia, Slovenia and Bulgaria. The asymptotic relationship between the nitrogen concentrations in mosses and EMEP modelled nitrogen deposition (averaged per 50 km × 50 km grid) across Europe showed less scatter when there were at least five moss sampling sites per grid. Factors potentially contributing to the scatter are discussed. In Switzerland, a strong (r(2) = 0.91) linear relationship was found between the total nitrogen concentration in mosses and measured site-specific bulk nitrogen deposition rates. The total nitrogen concentrations in mosses complement deposition measurements, helping to identify areas in Europe at risk from high nitrogen deposition at a high spatial resolution.

Mosses as biomonitors of atmospheric heavy metal deposition: spatial patterns and temporal trends in Europe.

In recent decades, mosses have been used successfully as biomonitors of atmospheric deposition of heavy metals. Since 1990, the European moss survey has been repeated at five-yearly intervals. Although spatial patterns were metal-specific, in 2005 the lowest concentrations of metals in mosses were generally found in Scandinavia, the Baltic States and northern parts of the UK; the highest concentrations were generally found in Belgium and south-eastern Europe. The recent decline in emission and subsequent deposition of heavy metals across Europe has resulted in a decrease in the heavy metal concentration in mosses for the majority of metals. Since 1990, the concentration in mosses has declined the most for arsenic, cadmium, iron, lead and vanadium (52-72%), followed by copper, nickel and zinc (20-30%), with no significant reduction being observed for mercury (12% since 1995) and chromium (2%). However, temporal trends were country-specific with sometimes increases being found.

Use of native mosses as biomonitors of heavy metals and nitrogen deposition in the surroundings of two steel works.

A biomonitoring survey using the moss species Hypnum cupressiforme Hedw. was conducted in the surroundings of two steel plants located in the North of Spain. Levels of V, Cr, Ni, Cu, Zn, As, Cd, Hg, Pb and N were determined. Very high concentrations in the areas of study were detected when compared to nearby unaffected regions. Similar trends were observed for all the elements in the differently orientated transects, showing an appreciable influence of the NW prevailing winds of the region in the dispersion of pollutants, as well as a clear decreasing gradient in the concentrations of metals in mosses within a distance of 1500 meters from the facilities. A differentiation between the elements emitted by the chimney as result of the industrial activity (V, Cr, Ni, Cu and As) and those with a high presence in steel slag deposits (Zn, Cd, Hg and Pb) was observed. The range of contamination was also established by means of the Contamination Factor, indicating a category 4 out of 6 categories, which shows the high levels reported in the areas of study. A different dynamic was registered for nitrogen regarding the rest of the heavy metals analysed except for Hg, probably due to the elevated volatility and mobility of both elements, as well as their high persistence in the atmosphere.

Ambient air levels of volatile organic compounds (VOC) and nitrogen dioxide (NO2) in a medium size city in Northern Spain.

Ambient concentrations of volatile organic compounds (VOC) and nitrogen dioxide (NO2) were measured by means of passive sampling at 40 sampling points in a medium-size city in Northern Spain, from June 2006 to June 2007. VOC and NO2 samplers were analysed by thermal desorption followed by gas chromatography/mass-selective detector and by visible spectrophotometry, respectively. Mean concentrations of benzene, toluene, ethylbenzene, xylenes, propylbenzene, trimethylbenzenes, and NO(2) were 2.84, 13.26, 2.15, 6.01, 0.59, 1.32 and 23.17 microg m(-3) respectively, and found to be highly correlated. Their spatial distribution showed high differences in small distances and pointed to traffic as the main emission source of these compounds. The lowest levels of VOC and NO2 occurred during summer, owing to the increase in solar radiation and to lower traffic densities. Mean concentrations of benzene and NO2 exceeded the European limits at some of the monitored points.

The Pb-hyperaccumulator aquatic fern Salvinia minima Baker, responds to Pb(2+) by increasing phytochelatins via changes in SmPCS expression and in phytochelatin synthase activity.

The relationship between accumulation of Pb(2+) and the activation of chelation and metal sequestration mechanisms mediated by phytochelatins (PC) was analyzed in the Pb(2+) hyperaccumulator aquatic fern Salvinia minima, after exposure to 40microM Pb(NO(3))(2). The tissue accumulation pattern of lead and the phytochelatin biosynthesis responses were analyzed in both, S. minima submerged root-like modified fronds (here named "roots"), and in its aerial leaf-like fronds ("leaves"). S. minima roots accumulated a significantly higher concentrations of Pb(+2) than leaves did. Accumulation of Pb(2+) in roots was bi-phasic with a first uptake phase reached after 3h exposure and a second higher uptake phase reached after 24h exposure. In leaves, a single delayed, smaller uptake phase was attained only after 9h of exposure. In roots lead accumulation correlated with an increased phytochelatin synthase (PCS) activity and an enhanced PC production. A higher proportion of polymerized PC(4) was observed in both tissues of exposed S. minima plants relative to unexposed ones, although a higher concentration of PC(4) was found in roots than in leaves. PCS activity and Pb(2+) accumulation was also higher in roots than in leaves. The expression levels of the S. minima PCS gene (SmPCS), in response to Pb(2+) treatment, were also evaluated. In S. minima leaves, the accumulation of Pb(2+) correlated with a marked increase in expression of SmPCS, suggesting a transcriptional regulation in the PCS activation and PC accumulation in this S. minima tissue. However, in roots, the basal expression of SmPCS was down-regulated after Pb(2+) treatment. This fact did not correlate with the later but strong increase in both, PCS activity and PC production; suggesting that the PC biosynthesis activation in S. minima roots occurs only by post-translational activation of PCS. Taken together, our data suggest that the accumulation of PC in S. minima is a direct response to Pb(2+) accumulation, and phytochelatins do participate as one of the mechanism to cope with Pb(2+) of this Pb-hyperaccumulator aquatic fern.