Shaping mechanisms of metal specificity in a family of metazoan Metallothioneins: evolutionary differentiation of Mollusc MTs.

Background: The degree of metal binding specificity in metalloproteins such as metallothioneins (MTs) can be crucial for their functional accuracy. Unlike most other animal species, pulmonate molluscs possess homometallic MT isoforms loaded with Cu+ or Cd2+. They have, so far, been obtained as native metal-MT complexes from snail tissues, where they are involved in the metabolism of the metal ion species bound to the respective isoform. However, it has not as yet been discerned if their specific metal occupation is the result of a rigid control of metal availability, or isoform expression programming in the hosting tissues or of structural differences of the respective peptides determining the coordinative options for the different metal ions. In this study, the Roman snail (Helix pomatia) Cu-loaded and Cd-loaded isoforms (HpCuMT and HpCdMT) were used as model molecules in order t o elucidate the biochemical and evolutionary mechanisms permitting pulmonate MTs to achieve specificity for their cognate metal ion. Results: HpCuMT and HpCdMT were recombinantly synthesized in the presence of Cd2+, Zn2+ or Cu2+ and corresponding metal complexes analysed by electrospray mass spectrometry and circular dichroism (CD) and ultra violet-visible (UV-Vis) spectrophotometry. Both MT isoforms were only able to form unique, homometallic and stable complexes (Cd6-HpCdMT and Cu12-HpCuMT) with their cognate metal ions. Yeast complementation assays demonstrated that the two isoforms assumed metal-specific functions, in agreement with their binding preferences, in heterologous eukaryotic environments. In the snail organism, the functional metal specificity of HpCdMT and HpCuMT was contributed by metal-specific transcription programming and cell-specific expression. Sequence elucidation and phylogenetic analysis of MT isoforms from a number of snail species revealed that they possess an unspecific and two metal-specific MT isoforms, whose metal specificity was achieved exclusively by evolutionary modulation of non-cysteine amino acid positions. Conclusion: The Roman snail HpCdMT and HpCuMT isoforms can thus be regarded as prototypes of isoform families that evolved genuine metal-specificity within pulmonate molluscs. Diversification into these isoforms may have been initiated by gene duplication, followed by speciation and selection towards opposite needs for protecting copper-dominated metabolic pathways from nonessential cadmium. The mechanisms enabling these proteins to be metal-specific could also be relevant for other metalloproteins.

Formation and distribution of point defects on a disclination line near a free nematic interface

Point defects of opposite signs can alternately nucleate on the -1/2 disclination line that forms near the free surface of a confined nematic liquid crystal. We show the existence of metastable configurations consisting of periodic repetitions of such defects. These configurations are characterized by a minimal interdefect spacing that is seen to depend on sample thickness and on an applied electric field. The time evolution of the defect distribution suggests that the defects attract at small distances and repel at large distances.

The role of dynamical polarization of the ligand-to-metal charge transfer excitations in the ab initio determination of effective exchange parameters

The role of the bridging ligand on the effective Heisenberg coupling parameters is analyzed in detail. This analysis strongly suggests that the ligand-to-metal charge transfer excitations are responsible for a large part of the final value of the magnetic coupling constant. This permits us to suggest a variant of the difference dedicated configuration interaction (DDCI) method, presently one of the most accurate and reliable for the evaluation of magnetic effective interactions. This method treats the bridging ligand orbitals mediating the interaction at the same level than the magnetic orbitals and preserves the high quality of the DDCI results while being much less computationally demanding. The numerical accuracy of the new approach is illustrated on various systems with one or two magnetic electrons per magnetic center. The fact that accurate results can be obtained using a rather reduced configuration interaction space opens the possibility to study more complex systems with many magnetic centers and/or many electrons per center.

First-principles study of the neutral molecular metal Ni(tmdt)2.

The electronic structure of the molecular solid Ni(tmdt)2, the only well characterized neutral molecular metal to date, has been studied by means of first-principles density functional calculations. It is shown that these calculations correctly describe the metallic vs semiconducting behavior of molecular conductors of this type. The origin of the band overlap leading to the metallic character and the associated Fermi surfaces has been studied.

Chiroptical measurement of chiral aggregates at liquid-liquid interface in centrifugal liquid membrane cell by Mueller matrix and conventional circular dichroism methods

The centrifugal liquid membrane (CLM) cell has been utilized for chiroptical studies of liquid-liquid interfaces with a conventional circular dichroism (CD) spectropolarimeter. These studies required the characterization of optical properties of the rotating cylindrical CLM glass cell, which was used under the high speed rotation. In the present study, we have measured the circular and linear dichroism (CD and LD) spectra and the circular and linear birefringence (CB and LB) spectra of the CLM cell itself as well as those of porphyrine aggregates formed at the liquid-liquid interface in the CLM cell, applying Mueller matrix measurement method. From the results, it was confirmed that the CLM-CD spectra of the interfacial porphyrin aggregates observed by a conventional CD spectropolarimeter should be correct irrespective of LD and LB signals in the CLM cell.

Modeling storm events to investigate the influence of the stream¿catchment interface zone on stream biogeochemistry

We formulate a new mixing model to explore hydrological and chemical conditions under which the interface between the stream and catchment interface (SCI) influences the release of reactive solutes into stream water during storms. Physically, the SCI corresponds to the hyporheic/riparian sediments. In the new model this interface is coupled through a bidirectional water exchange to the conventional two components mixing model. Simulations show that the influence of the SCI on stream solute dynamics during storms is detectable when the runoff event is dominated by the infiltrated groundwater component that flows through the SCI before entering the stream and when the flux of solutes released from SCI sediments is similar to, or higher than, the solute flux carried by the groundwater. Dissolved organic carbon (DOC) and nitrate data from two small Mediterranean streams obtained during storms are compared to results from simulations using the new model to discern the circumstances under which the SCI is likely to control the dynamics of reactive solutes in streams. The simulations and the comparisons with empirical data suggest that the new mixing model may be especially appropriate for streams in which the periodic, or persistent, abrupt changes in the level of riparian groundwater exert hydrologic control on flux of biologically reactive fluxes between the riparian/hyporheic compartment and the stream water.

Metal complexation by electroanalytical techniques: hard- and soft-modelling approaches

En la investigació de la complexació de metalls mitjançant eines electroanalítiques són emprades dues aproximacions generals. La primera, anomenada de modelatge dur (hardmodelling), es basa en la formulació d'un model fisicoquímic conjunt per als processos electròdic i de complexació i en la resolució analítica o numèrica del model. Posteriorment, l'ajust dels paràmetres del model a les dades experimentals donarà la informació desitjada sobre el procés de complexació. La segona aproximació, anomenada de modelatge tou (soft-modelling), es basa en la identificació d'un model de complexació a partir de l'anàlisi numèrica i estadística de les dades, sense cap assumpció prèvia d'un model. Aquesta aproximació, que ha estat extensivament emprada amb dades espectroscòpiques, ho ha estat poquíssim amb dades electroquímiques. En aquest article tractem de la formulació d'un model (hard-modelling) per a la complexació de metalls en sistemes amb mescles de lligands, incloent-hi lligands macromoleculars, i de l'aplicació d

Heavy metal accumulation by intestinal helminths of vertebrates

The relevancy of parasites as potential indicators of environmental quality has been increasing over the last years, mostly due to the variety of ways in which they respond to anthropogenic pollution. The use of fish parasites as bioindicators of heavy metal pollution in aquatic ecosystems has been widely studied. However, little information concerning terrestrial habitats is presently available. In fact, in the last two decades several studies have been performed worldwide in different habitats and/or conditions (theoretically both in polluted and unpolluted terrestrialecosystems, but mainly in aquatic ecosystems) in order to investigate heavy metal pollution using parasitological models. Different groups of vertebrates (mainly fish, mammals and birds) and several parasitological models have been tested involving acanthocephalans mostly, but also cestodes and nematodes. It is not the aim of this chapter to do a complete revision of the availabledata concerning this subject. Instead, we emphasize some general aspects and compile a mini-review of the work performed in this field by our research group. The results obtained until now allow confirming several parasitic models as promising bioindicator systems to evaluate environmental cadmium and mainly lead pollution in terrestrial non-urban habitats, as it was already demonstrated for aquatic ecosystems. The present knowledge also allows confirming that parasites can reveal environmental impact. Environmental parasitology is an interdisciplinary field, which needs simultaneous expertise from toxicology, environmental chemistry and parasitology. Furthermore, environmental parasitology should be taken into account in order to increase the efficiency of environmental monitoring programs.

Heavy metal accumulation by intestinal helminths of vertebrates

The relevancy of parasites as potential indicators of environmental quality has been increasing over the last years, mostly due to the variety of ways in which they respond to anthropogenic pollution. The use of fish parasites as bioindicators of heavy metal pollution in aquatic ecosystems has been widely studied. However, little information concerning terrestrial habitats is presently available. In fact, in the last two decades several studies have been performed worldwide in different habitats and/or conditions (theoretically both in polluted and unpolluted terrestrialecosystems, but mainly in aquatic ecosystems) in order to investigate heavy metal pollution using parasitological models. Different groups of vertebrates (mainly fish, mammals and birds) and several parasitological models have been tested involving acanthocephalans mostly, but also cestodes and nematodes. It is not the aim of this chapter to do a complete revision of the availabledata concerning this subject. Instead, we emphasize some general aspects and compile a mini-review of the work performed in this field by our research group. The results obtained until now allow confirming several parasitic models as promising bioindicator systems to evaluate environmental cadmium and mainly lead pollution in terrestrial non-urban habitats, as it was already demonstrated for aquatic ecosystems. The present knowledge also allows confirming that parasites can reveal environmental impact. Environmental parasitology is an interdisciplinary field, which needs simultaneous expertise from toxicology, environmental chemistry and parasitology. Furthermore, environmental parasitology should be taken into account in order to increase the efficiency of environmental monitoring programs.

Somatostatin Subtype‑2 Receptor-Targeted Metal-Based Anticancer Complexes

Conjugates of a dicarba analogue of octreotide, a potent somatostatin agonist whose receptors are overexpressed on tumor cells, with [PtCl2(dap)] (dap = 1-(carboxylic acid)-1,2-diaminoethane) (3), [(η6-bip)Os(4-CO2-pico)Cl] (bip = biphenyl, pico = picolinate) (4), [(η6-p-cym)RuCl(dap)]+ (p-cym = p-cymene) (5), and [(η6-p-cym)RuCl(imidazole-CO2H)(PPh3)]+ (6), were synthesized by using a solid-phase approach. Conjugates 35 readily underwent hydrolysis and DNA binding, whereas conjugate 6 was inert to ligand substitution. NMR spectroscopy and molecular dynamics calculations showed that conjugate formation does not perturb the overall peptide structure. Only 6 exhibited antiproliferative activity in human tumor cells (IC50 = 63 ± 2 μM in MCF-7 cells and IC50 = 26 ± 3 μM in DU-145 cells) with active participation of somatostatin receptors in cellular uptake. Similar cytotoxic activity was found in a normal cell line (IC50 = 45 ± 2.6 μM in CHO cells), which can be attributed to a similar level of expression of somatostatin subtype-2 receptor. These studies provide new insights into the effect of receptor-binding peptide conjugation on the activity of metal-based anticancer drugs, and demonstrate the potential of such hybrid compounds to target tumor cells specifically.

Formation of disclinations line near a free nematic interface

We have studied the nucleation and the physical properties of a -1/2 wedge disclination line near the free surface of a confined nematic liquid crystal. The position of the disclination line has been related to the material parameters (elastic constants, anchoring energy, and favored anchoring angle of the molecules at the free surface). The use of a planar model for the structure of the director field (whose predictions have been contrasted to those of a fully three-dimensional model) has allowed us to relate the experimentally observed position of the disclination line to the relevant properties of the liquid crystals. In particular, we have been able to observe the collapse of the disclination line due to a temperature-induced anchoring-angle transition, which has allowed us to rule out the presence of a real disclination line near the nematic/isotropic front in directional growth experiments.

Structures of ethylenediaminetetraacetato (3-) metal complexes. I complexes with Co, Mg and Cd metals

(I): Hexaaquacobalt(II) aqua[ethylenediaminetetraacetato(3-)]cobaltate(II) dihydrate, [Co(H2O)6][Co(C10H13N2O8)(H2O)]2.2H2O (Ibis): Hexaaquamagnesium(II) aqua[ethylenediaminetetraacetato(3-)]magnesiate(II) dihydrate, [Mg(H2O)6][Mg(C10H13N2O8)(H2O)]2.2H2O (II):Tetraaquabis{aqua[ethylenediaminetetraacetato(3-)]cadmium(II)-O-O'}Cadmium(II) tetrahydrate

Structures of ethylenediaminetetraacetato (3-) metal complexes. I complexes with Co, Mg and Cd metals

(I): Hexaaquacobalt(II) aqua[ethylenediaminetetraacetato(3-)]cobaltate(II) dihydrate, [Co(H2O)6][Co(C10H13N2O8)(H2O)]2.2H2O (Ibis): Hexaaquamagnesium(II) aqua[ethylenediaminetetraacetato(3-)]magnesiate(II) dihydrate, [Mg(H2O)6][Mg(C10H13N2O8)(H2O)]2.2H2O (II):Tetraaquabis{aqua[ethylenediaminetetraacetato(3-)]cadmium(II)-O-O'}Cadmium(II) tetrahydrate

Structures of ethylenediaminetetraacetato (3-) metal complexes. I complexes with Co, Mg and Cd metals

(I): Hexaaquacobalt(II) aqua[ethylenediaminetetraacetato(3-)]cobaltate(II) dihydrate, [Co(H2O)6][Co(C10H13N2O8)(H2O)]2.2H2O (Ibis): Hexaaquamagnesium(II) aqua[ethylenediaminetetraacetato(3-)]magnesiate(II) dihydrate, [Mg(H2O)6][Mg(C10H13N2O8)(H2O)]2.2H2O (II):Tetraaquabis{aqua[ethylenediaminetetraacetato(3-)]cadmium(II)-O-O'}Cadmium(II) tetrahydrate