Francisco Martín Romero

Intracellular pH in sperm physiology

Intracellular pH (pHi) regulation is essential for cell function. Notably, several unique sperm ion transporters and enzymes whose elimination causes infertility are either pHi dependent or somehow related to pHi regulation. Amongst them are: CatSper, a Ca(2+) channel; Slo3, a K(+) channel; the sperm-specific Na(+)/H(+) exchanger and the soluble adenylyl cyclase. It is thus clear that pHi regulation is of the utmost importance for sperm physiology. This review briefly summarizes the key components involved in pHi regulation, their characteristics and participation in fundamental sperm functions such as motility, maturation and the acrosome reaction.

The role of cassiterite controlling arsenic mobility in an abandoned stanniferous tailings impoundment at Llallagua, Bolivia.

The surface water contamination by potentially toxic elements (PTE) leached from mine tailings is a major environmental concern. However, the formation of insoluble solid phases can control the mobility of PTE, with subsequent decrease of the risk that tailings suppose to the environment. We characterized the tailings from a tin inactive mine in Llallagua, Bolivia in order to assess the risk for surface water quality. These tailings contain high concentrations of PTE, with up to 94,344 mg/kg Fe, 9,135 mg/kg Sn, 4,606 mg/kg As, 1,362 mg/kg Cu, 1,220 mg/kg Zn, 955 mg/kg Pb and 151 mg/kg Cd. Oxidation of sulfide minerals in these tailings generates acid leachates (pH=2.5-3.5), rich in SO4(2-) and dissolved PTE, thereby releasing contaminants to the surface waters. Nevertheless, the concentrations of dissolved Sn, As and Pb in acid leachates are low (Sn<0.01 mg/L; As=0.25-2.55 mg/L; Pb<0.05 mg/L). This indicates that, for the most part, Sn, As and Pb are being retained by the solid phases in the impoundment, so that these elements are not reaching the surface waters. Fe-bearing cassiterite-an insoluble and weathering-resistant oxide mineral-is abundant in the studied tailing deposits; it should be the main solid phase controlling Sn and As mobility in the impoundment. Additionally, jarosite and plumbojarosite, identified among the secondary minerals, could also play an important role controlling the mobility of As and Pb. Taking into account (a) the low solubility constants of cassiterite (Ksp=10(-64.2)), jarosite (Ksp=10(-11)) and plumbojarosite (Ksp=10(-28.66)), and (b) the stability of these minerals under acidic conditions, we can conclude that they control the long-term fate of Sn, As and Pb in the studied tailings.

Metal accumulation by plant species growing on a mine contaminated site in Mexico

The present work aims to assess the efficiency of heavy metal accumulation of native species growing in contaminated soils in the mining district of Taxco, Mexico. Soil and tailing sampling was conducted in three study sites: La Concha, El Fraile, and a control site. The study localities present diverse metal concentrations with significant differences in their proportion in the geochemical fractions. Results show that species Cupressus lindleyi and Juniperus deppeana accumulate Zn and Mn in anomalous concentrations at La Concha, where Zn is present in soluble fractions. Manganese, despite not being present mostly in the soluble fraction in soils and tailings, seems to have been increased in the soluble fraction after the plant growth. In contrast, samples of the same species taken at El Fraile and in the control site, where Zn and Mn are mainly contained in the residual fraction, do not show an anomalous enrichment. Other analyzed species growing under the same contamination conditions in La Concha (Jacaranda mimosifolia and Psidium guajava) do not show anomalous concentrations. These facts confirm the Zn and Mn accumulation capacity of C. lindleyi and Ju. deppeana, which depends on their accumulation ability and on the concentration of these elements in the soluble fraction.

Fractionation and mobility of thallium in areas impacted by mining-metallurgical activities: Identification of a water-soluble Tl(I) fraction

Mining and metallurgy generate residues that may contain thallium (Tl), a highly toxic metal, for which it is currently not feasible to determine its geochemical speciation through X-ray absorption spectroscopy due to a combination of very low contents and the interference of accompanying high arsenic contents. Therefore, fractionation studies in residues and soils are required to analyze the mobility and bioavailability of this metal, which in turn provide information to infer its speciation. For this purpose, in this work a modification of the BCR procedure was applied to residues and contaminated soils from three mining zones of Mexico and two mining zones of Spain, spanning samples with acidic to alkaline pH values. The Tl extraction procedure consisted of the following fractions: (1) water-extractable, (2) easily exchangeable and associated to carbonates, associated to (3) poorly-crystalline and (4) crystalline Fe and Mn oxyhydroxides, and (5) associated to organic matter and sulfides; and finally a residual fraction as associated to refractory primary and other secondary minerals. The extracted contents were analyzed by Inductively-Coupled Plasma with Mass Spectrometry. Surprisingly, water-soluble, in Tl(I) oxidation state, was detected in most areas, regardless of the pH, a fact that has not been reported before in these environments, and alerts to potential health risks not previously identified. Most of the samples from a metallurgy area showed high levels of Tl in non-residual fractions and a strong correlation was obtained between extracted Mn and Tl in the third fraction, suggesting its association to poorly crystalline manganese oxides. In the majority of samples from purely mining environments, most of the Tl was found in the residual fraction, most probably bound to alumino-silicate minerals. The remaining Tl fractions were extracted mainly associated to the reducible mineral fractions, and in one case also in the oxidizable fraction (presumably associated to sulfides). Capsule: Soluble Tl(I) was found in all soil samples contaminated with either mining or metallurgical wastes. Additionally, in those affected by metallurgical wastes a very strong Tl-Mn correlation was found.

Plants from the abandoned Nacozari mine tailings: evaluation of their phytostabilization potential

Phytostabilization is a remediation technology that uses plants for in-situ stabilization of contamination in soils and mine tailings. The objective of this study was to identify native plant species with potential for phytostabilization of the abandoned mine tailings in Nacozari, Sonora in northern Mexico. A flora of 42 species in 16 families of angiosperms was recorded on the tailings site and the abundance of the most common perennial species was estimated. Four of the five abundant perennial species showed evidence of regeneration: the ability to reproduce and establish new seedlings. A comparison of selected physicochemical properties of the tailings in vegetated patches with adjacent barren areas suggests that pH, electrical conductivity, texture, and concentration of potentially toxic elements do not limit plant distribution. For the most abundant species, the accumulation factor for most metals was <1, with the exception of Zn in two species. A short-term experiment on adaptation revealed limited evidence for the formation of local ecotypes in Prosopis velutina and Amaranthus watsonii. Overall, the results of this study indicate that five native plant species might have potential for phytostabilization of the Nacozari tailings and that seed could be collected locally to revegetate the site. More broadly, this study provides a methodology that can be used to identify native plants and evaluate their phytostabilization potential for similar mine tailings.

CHARACTERIZATION OF DISSOLVED ORGANIC MATTER IN AN AGRICULTURAL WASTEWATER-IRRIGATED SOIL, IN SEMI ARID MEXICO

Most agricultural soils in semi-arid regions present a deficiency of organic matter (SOM). In order to improve this soil, wastewater is used due to its high organic carbon content. The objective of the present work was to characterize the dissolved organic matter in five residual water samples and ten soil samples from the agricultural area of the municipality of San Luis Potosi by 3D fluorescence spectroscopy. The changes in some physical and chemical properties of the soil were also analyzed. The soil samples were collected at different depths to observe the anthropogenic organic matter presence, their retention in the soil profile, their fluorescence intensity changes and their migration into the aquifer. Temperature, electrical conductivity, dissolved oxygen, oxidation- reduction potential and total dissolved solids were determined in situ. The major anions and cations: Na + , K + , Mg 2+ , Ca 2+ , PO 4 3– , SO 4 2– , NO 3– , NO 2 – , were analysed in laboratory. The results show the great contribution of organic matter (>2020 mg/L) in the residual water used for irrigation, as well as, the low content of SOM. The physical and chemical results indicate that the high electrical conductivity of the water, represents a risk of salinization for soils, but not of short term sodicity. The 3D fluorescence spectra of the soil profile, shows the presence of humic and fulvic acids, aromatic proteins and products of microbial degradation. The latter observed in that depth where there is a greater percentage of clay.

Effect of an acid mine spill on soils in Sonora River Basin: Micromorphological indicators

The role of soil could be decisive for the neutralization of the acid solution and immobilization of the metallic contaminants produced by mining industry. We studied micromorphological indicators of the interaction between the acid solution and soil material in the profile of Fluvisol at the Bacanuchi river (Sonora River Basin, Mexico) terrace affected by the catastrophic mine spill and in analyzing sample from a soil column treated with an acid liquid imitating the spill. Original unaffected soil is sandy with poorly developed pedogenetic features, however, frequent primary and secondary micritic and sparitic carbonates define high pH values. In the soil influenced by acid solutions under natural and laboratory conditions, carbonates were absent whereas neoformed gypsum crystals with radial intergrowth were observed together with accumulation of fine material enriched in ferruginous pigment. Micromorphometric quantification of the iron-rich fine material has shown its increase after interaction with the acid solution. We conclude that the interaction consisted of the neutralization reaction between the more reactive phases of the soil and the acid solution during which carbonates were consumed and gypsum was neoformed. Fine iron oxides precipitated after neutralization of acidity, the pH increased and the color changed. It is highly probable that other metallic contaminants co-precipitated with the ferruginous components.

Source apportionment and environmental fate of lead chromates in atmospheric dust in arid environments

The environmental fate of lead derived from traffic paint has been poorly studied in developing countries, mainly in arid zones. For this purpose, a developing city located in the Sonoran desert (Hermosillo, Mexico), was chosen to conduct a study. In this paper the lead chromate (crocoite) sources in atmospheric dust were addressed using a combination of Raman microspectroscopy, X-ray diffraction, scanning electron microscopy (SEM), and Pb isotope measurements. A high concentration of Pb and Cr as micro- and nanostructured pigments of crocoite is reported in yellow traffic paint (n=80), road dust (n=146), settled dust in roofs (n=21), and atmospheric dust (n=20) from a developing city located in the Sonoran Desert. 10 samples of peri-urban soils were collected for local geochemical background. The paint photodegradation and erosion of the asphaltic cover are enhanced by the climate, and the presence of the mineral crocoite (PbCrO4) in road dust with an aerodynamic diameter ranging from 100nm to 2μm suggests its integration into the atmosphere by wind resuspension processes. A positive PbCr correlation (R2=0.977) was found for all studied samples, suggesting a common source. The Pb-isotope data show signatures in atmospheric dust as a product of the mixing of two end members: i) local soils and ii) crocoite crystals as pigments in paint. The presence of lead chromates in atmospheric dust has not been previously documented in Latin America, and it represents an unknown health risk to the exposed population because the identified size of crystals can reach the deepest part of lungs.

Robust evaluation of intermolecular FRET using a large Stokes shift fluorophore as a donor

Fluorescence (or Förster) resonance energy transfer (FRET) is a straightforward and sensitive technique to evaluate molecular interactions. However, most of the popular FRET pairs suffer cross-excitation of the acceptor, which could lead to false positives. To overcome this problem, we selected a large Stokes shift (LSS) fluorophore as a FRET donor. As a successful example, we employed a new FRET pair mAmetrine (an LSS yellow fluorescence protein)/DY-547 (a cyanine derivative) to substitute CFP/fluorescein that we previously employed to study molecular interactions between cyclic nucleotide-binding domains and cyclic nucleotides. The new FRET pair is practically free of cross-excitation of the acceptor. Namely, a change in the fluorescence spectral shape implies evidence of FRET without other control experiments.

FRET‐based binding assay between a fluorescent cAMP analogue and a cyclic nucleotide‐binding domain tagged with a CFP

The cyclic nucleotide‐binding domain (CNBD) functions as a regulatory domain of many proteins involved in cyclic nucleotide signalling. We developed a straightforward and reliable binding assay based on intermolecular fluorescence resonance energy transfer (FRET) between an adenosine‐3′, 5′‐cyclic monophosphate analogue labelled with fluorescein and a recombinant CNBD of human EPAC1 tagged with a cyan fluorescence protein (CFP). The high FRET efficiency of this method (~ 80%) allowed us to perform several types of binding experiments with nanomolar range of sample using conventional equipment. In addition, the CFP tag on the CNBD enabled us to perform a specific binding experiment using an unpurified protein. Considering these advantages, this technique is useful to study poorly characterized CNBDs.