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.

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.

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.

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.

[Diseases in heroin addiction]. / Patologías médicas en la adicción a la heroína.

An analysis of the medical pathologies seen in heroin addicts heroin addicts hospitalized in the Provincia de Vizcaya, Basque Country, Spain, from 1982 to 1987, and its possible association to the infection by the human immunodeficiency virus, HIV, is presented. An increasing of hospitalized addicts per year was observed, being infectious diseases the main pathology detected. HIV infection was present in more than 90% of individuals with non opportunistic acquired infections. Significant annual increase in the number of tuberculosis and candida infections was present. The possible association of HIV infection with increased susceptibility to commonly acquired non opportunistic infections is discussed.