Applications of nuclear analytical techniques in environmental research. Plenary lecture.

Among nuclear analytical techniques, neutron activation analysis (NAA) is particularly useful for environmental studies. It affords low detection limits for many elements, high specificity and few sources of systematic error, which means that high accuracy is attainable. Neutron activation analysis is particularly useful for trace and ultra-trace analysis of environmental samples (water, soils, rocks and biological material). In trace element work associated with pollution, instrumental NAA is a powerful technique for multi-element surveys, in particular when combined with other spectroscopic techniques. Nuclear techniques, as with most analytical techniques, cannot be used to distinguish between different physico-chemical forms of an element per se. When used in combination with appropriate separation techniques, however, nuclear techniques can provide valuable information about trace element speciation in environmental and biological systems. From dynamic tracer experiments, i.e., addition of chemically well defined labelled compounds to environmental systems, valuable information can be obtained on the distribution of species and on microchemical processes influencing the physico-chemical forms. In these laboratories, speciation studies on trace elements in natural waters have been carried out by using instrumental NAA in combination with physical separation techniques, such as dialysis and ultrafiltration, in situ and in the laboratory. Dynamic radiotracer experiments have provided important information about processes influencing the speciation of trace elements in aquatic systems. Sequential extraction techniques have proved to be useful in studies on sediments and soils when combined with NAA. Sequential extractions also provide significant information about the physico-chemical behaviour of radionuclides supplied to natural soils from the Chernobyl accident.     

Trace element analytical speciation in biological systems: importance,challenges and trends

Speciation of trace elements is a relatively new field and it was in toxicology that the relationship between the chemical form of a metal and its harmful effects was first recognized. The present need for chemical speciation information in biochemistry bioinorganic and clinical chemistry is documented in an attempt to justify the present demand for innovative chemical speciation strategies and analytical technologies.The challenge and complexity of speciation is stressed and three different categories of analytical speciation of increasing analytical difficulty are proposed. Analytical strategies developed so far to try to tackle speciation problems (computational approaches, direct species-specific and hybrid techniques) are reviewed and critically assessed for biological materials. It is indisputable these days that in most cases of real-life analytical speciation we have to resort to the development and use of hybrid techniques combining an adequate separation technique for the species physical separation and an element specific detector such as those based in atomic spectrometry. Examples of such strategies, as developed mainly in the author's laboratory and including chromatographic and non-chromatographic type hybrid strategies coupled to flame, plasma and electrothermal vaporization atomic detectors, are discussed in more detail.Finally, in light of the latest trends observed in this new field, the author attempts to cast a forward look into the foreseeable future of analytical speciation research in biological and biomedical sciences. The urgent plea for quality assurance in non-routine analysis and the concept of using complementary analytical techniques and definitive methods to attack the complexity of chemical speciation in biological systems are particularly highlighted.     

Conformationally rigid proteomimetics: a case study in designing antimicrobial aryl oligomers

The promise of proteomics to provide a vast library of protein structural data is exciting to scientists desiring an unprecedented understanding of the relationship between protein structure and function. This powerful knowledge will provide insight into the design rules for proteomimetics which are oligomers and polymers that can be more stable and inexpensive to produce than natural proteins, but still emulate the main biological function of the natural molecule. This Emerging Area article is intended to stimulate discussion on innovative strategies to design the next generation of proteomimetics. Specifically we will examine the design evolution of facially amphiphilic aryl oligomers, compounds that act as synthetic mimics of antimicrobial peptides (SMAMPs) and are known to interact with lipid bilayers. An increasingly important goal in the field of antimicrobial polymers is to develop strategies to rationally design membrane-binding SMAMPs, that are highly cell-selective, from any preferred backbone and molecular weight. It is expected that lessons learned from studying these oligomers can be applied to other systems where mimics are desired to interact with extended surfaces and where it would be most productive to consider mimicking the protein of interest with a large molecule. Obvious examples include disrupting protein-protein interactions or binding long tracts of DNA to control gene expression.     

Osmolarity effects on red blood cell elution in sedimentation field-flow fractionation.

Field-flow fractionation (FFF) is an analytical technique particularly suitable for the separation, isolation, and characterization of macromolecules and micrometer- or submicrometer-sized particles. This chromatographic-like methodology can modulate the retention of micron-sized species according to an elution mode described to date as "steric hyperlayer". In such a model, differences in sample species size, density, or other physical parameters make particle selective elution possible depending on the configuration and the operating conditions of the FFF system. Elution characteristics of micron-sized particles of biological origin, such as cells, can be modified using media and carrier phases of different osmolarities. In these media, a cells average size, density, and shape are modified. Therefore, systematic studies of a single reference cell population, red blood cells (RBCs), are performed with 2 sedimentation FFF systems using either gravity (GrFFF) or a centrifugational field (SdFFF). However, in all cases, normal erythrocyte in isotonic suspension elutes as a single peak when fractionated in these systems. With carrier phases of different osmolarities, FFF elution characteristics of RBCs are modified. Retention modifications are qualitatively consistent with the "steric-hyperlayer" model. Such systematic studies confirm the key role of size, density, and shape in the elution mode of RBCs in sedimentation FFF for living, micronsized biological species. Using polymers as an analogy, the RBC population is described as highly "polydisperse". However, this definition must be reconsidered depending on the parameters under concern, leading to a matricial concept: multipolydispersity. It is observed that multipolydispersity modifications of a given RBC population are qualitatively correlated to the eluted sample band width.     

Current perspectives in analyte extraction strategies for tin and arsenic speciation

Nowadays, reliable and robust detectors can be considered standard laboratory instrumentation, which, for most of the elements provide quantitation limits in the lower ng/g range. Despite these advances in detector technology, sample preparation is by far the most important error source in modern analytical method development and can be judged as the "Achilles' heel" of any analytical process regarding reliability of the obtained results and time consumption. The aim of the present review is to highlight modern trends for tin and arsenic speciation, as these analytes can be considered as models for challenges in modern method development in this field. First background information, legislative aspects and current needs are elucidated. Then the role of sample treatment within the process of method development in speciation is discussed, followed by a presentation of modern extraction techniques, matching the requirements for arsenic and tin speciation analysis: to provide mild conditions in order to ensure species preservation, to improve species recovery, to enhance sample throughput and to be suitable for hyphenation with chromatographic separation systems. The review includes applications on tin and arsenic speciation, covering the period of 2001-2006.     

A bone ash standard for90Sr,210Pb,210Po,uranium and the actinides

Possibilities and problems of experimental differentiation of two chemical or particle forms of element M (Acentral atom or group, Bcomplex, metalorganic or bioinorganic species, colloid, etc.) between which mutual equilibria can exist at their partition in two-phase systems, when the gross (analytical) concentrations are determined in each phase (by AAS, NAA, or radiometrically) are outlined for: (1) static distribution (batch separation) between two immiscible phases, when both the partial concentrations of A and B and their distribution coefficients generally can be determined by a single or fractional separation, (2) dynamic distribution (chromatographic separation) between the peaks of two metal species, when the potentialities can be strongly affected by secondary equilibria or when the sample-eluent interactions can be assessed.     

Approximate scheme for calculating van der Waals interactions between finite cylindrical volume elements

A successful approach to calculating van der Waals (vdW) forces between irregular bodies is to divide the bodies into small cylindrical volume elements and integrate the vdW interactions between opposing elements. In this context it has been common to use Hamaker's expression for parallel plates to approximate the vdW interactions between the opposing elements. This present study shows that Hamaker's vdW expression for parallel plates does not accurately describe the vdW interactions for co-axial cylinders having a ratio of cylinder radius to separation distance (R/D) of 10 or less. This restricts the systems that can be simulated using this technique and explicitly excludes consideration of topographical or compositional variations at the nanoscale for surfaces that are in contact or within a few nm of contact. To address this limitation, approximate analytical expressions for nonretarded vdW forces between finite cylinders in different orientations are derived and are shown to produce a high level of agreement with forces calculated using full numerical solutions of the corresponding Hamaker's equations. The expressions developed here allow accurate calculation of vdW forces in systems where particles are in contact or within a few nm of contact with surfaces and the particles and/or surfaces have heterogeneous nanoscale morphology or composition. These calculations can be performed at comparatively low computational cost compared to the full numerical solution of Hamaker's equations.     

New methods for nanotoxicology: synchrotron radiation-based techniques

Nanotoxicology, a new branch of bionanoscience, deals with the study and application of the toxic or biological effects of nanomaterials or nanostructures, and aims to fill gaps in our knowledge of interactions between nano- and biosystems. However, progress in this new discipline largely relies on developing methodology to characterize nanomaterials in biological samples, quantify nanoparticles in living systems, and study their uptake, translocation, biodistribution, location and chemical status in vitro and in vivo, etc. In this review article, we focus on the main features of synchrotron radiation-based methods and their application to the study of the toxicological activities of nanomaterials. Synchrotron radiation-based analytical techniques are shown to provide a potent means for characterizing the toxic or biological behaviors of nanoparticles in biological systems.     

Mapping of RNA-protein interactions

RNA-protein interactions are important biological events that perform multiple functions in all living organisms. The wide range of RNA interactions demands diverse conformations to provide contacts for the selective recognition of proteins. Various analytical procedures are presently available for quantitative analyses of RNA-protein complexes, but analytical-based mapping of these complexes is essential to probe specific interactions. In this overview, interactions of functional RNAs and RNA-aptamers with target proteins are discussed by means of mapping strategies.     

Molecular activation analysis for chemical species studies

Molecular activation analysis (MAA) refers to an activation analysis method that is able to provide information on the chemical species of elements in systems of interest, though its exact definition has yet to be assigned. Its development is strongly stimulated by the urgent need to know the chemical species of elements, because knowledge of bulk contents or concentrations is often insufficient to judge biological, environmental or geochemical effects of elements. The features, methodology and limitations of MAA are outlined, as well as the up-to-date MAA progress in our laboratory. Received: 14 August 1998 / Revised: 30 November 1998 / Accepted: 1 December 1998     

Classification of So-Called Non-Covalent Interactions Based on VSEPR Model

The variety of interactions have been analyzed in numerous studies. They are often compared with the hydrogen bond that is crucial in numerous chemical and biological processes. One can mention such interactions as the halogen bond, pnicogen bond, and others that may be classified as σ-hole bonds. However, not only σ-holes may act as Lewis acid centers. Numerous species are characterized by the occurrence of π-holes, which also may play a role of the electron acceptor. The situation is complicated since numerous interactions, such as the pnicogen bond or the chalcogen bond, for example, may be classified as a σ-hole bond or π-hole bond; it ultimately depends on the configuration at the Lewis acid centre. The disadvantage of classifications of interactions is also connected with their names, derived from the names of groups such as halogen and tetrel bonds or from single elements such as hydrogen and carbon bonds. The chaos is aggravated by the properties of elements. For example, a hydrogen atom can act as the Lewis acid or as the Lewis base site if it is positively or negatively charged, respectively. Hence names of the corresponding interactions occur in literature, namely hydrogen bonds and hydride bonds. There are other numerous disadvantages connected with classifications and names of interactions; these are discussed in this study. Several studies show that the majority of interactions are ruled by the same mechanisms related to the electron charge shifts, and that the occurrence of numerous interactions leads to specific changes in geometries of interacting species. These changes follow the rules of the valence-shell electron-pair repulsion model (VSEPR). That is why the simple classification of interactions based on VSEPR is proposed here. This classification is still open since numerous processes and interactions not discussed in this study may be included within it.     

Interaction of chemical species with biological regulation of the metabolism of essential trace elements

Variations in the chemical speciation of dietary trace elements can result in the provision of different amounts of these micronutrients to the organism and might thus induce interactions with trace-element metabolism. The chemical species of Zn, Fe, Cu, and Mn can interact with other components of the diet even before reaching the site of absorption, e.g. by formation of poorly soluble complexes with phytic acid. This might considerably modify the amount of metabolically available trace elements; differences between absorptive capacity per se toward dietary species seems to be less important. Homeostasis usually limits the quantities of Zn, Fe, Cu, and Mn transported from the gut into the organism, and differences between dietary species are largely eliminated at this step. There is no homeostatic control of absorption of Se and I, and organisms seem to be passively exposed to influx of these micronutrients irrespective of dietary speciation. Inside the organism the trace elements are usually converted into a metabolically recognizable form, channeled into their biological functions, or submitted to homeostatically controlled excretion. Some dietary species can, however, be absorbed as intact compounds. As long as the respective quantities of trace elements are not released from their carriers, they are not recognized properly by trace element metabolism and might induce tissue accumulation, irrespective of homeostatic control.     

Analytical methods for nano-bio interface interactions

Knowledge on the interactions between engineered nanomaterials (ENMs) and biological systems is critical both for the assessment of biological effects of ENMs and for the rational design of ENM-based products. However, probing the events that occur at the nano-bio interface remains extremely challenging due to their complex and dynamic nature. So far, the understanding of mechanisms underlying nano-bio interactions has been mainly limited by the lack of proper analytical techniques with sufficient sensitivity, selectivity and resolution for characterization of nano-bio interface events. Moreover, many classic bioanalytical methods are not suitable for direct measurement of nano-bio interface interactions. These have made establishing analytical methodologies for systematic and comprehensive study of nano-bio interface one of the most focused areas in nanobiology. In this review we have discussed some representative developments regarding analytical techniques for nano-bio interface characterization, including the improvements of traditional methods and the emergence of powerful new technologies. These developments have allowed ultrasensitive, real-time analysis of interactions between ENMs and biomolecules, transformations of ENMs in biological environment, and impacts of ENMs on living systems on molecular or cellular level.     

The role of element speciation in environmental and occupational medicine

In order to evaluate the interaction with the environment or to assess absorption, binding mechanisms, reactivity and excretion of elements in humans, element speciation can provide more information than the analysis of element as a whole. Some examples that confirm the importance of speciation depend on the choice of the most appropriate indicator or representative matrix. The determination of As(III), As(V), monomethylarsonic and dimethylarsinic acids can be used to evaluate occupational exposure to As. Exposure to inorganic Hg should be measured by its content in urine, whereas in the case of exposure to alkyl Hg, blood and hair should be considered. Speciation may also be useful in studying element toxicokinetics, since it is well known that hexavalent Cr is taken up more than the trivalent form, and that species of the same metal are differently partitioned in blood. Pentavalent forms of As are absorbed more than trivalent forms, and the organic species of elements are excreted faster than inorganic species. In addition, speciation can play an important role in assessing element toxicodynamics. The toxicity of the three oxidation states of Hg differs considerably; for As a decreasing toxicity from arsenite to dimethylarsinic acid is proposed; for organotin compounds, higher toxicity for ethyl groups than for phenyl groups is reported. However, speciation in biological media is difficult when applied to other elements because of the lack of information on the existence and significance of species whose determination could be valuable. Furthermore, there may be no analytical methods that allow an accurate measurement of the species. The feasibility of speciation in occupational and environmental medicine depends mainly on our capability to solve some problems related to the identification and determination of species and on the demonstration that species measurement represents a clear improvement compared to total element determination.     

Colloidal interactions at fluid interfaces

We discuss qualitative and quantitative aspects of the effective interactions between micrometer-sized colloids of different types trapped at fluid interfaces, with a particular emphasis on the relation between experimental and theoretical results. For colloids of that size, the interactions can broadly be classified into "direct" ones such as electrostatic, magnetic, or elastic ones. Such interactions appear also for colloids in bulk systems, but they are modified at interfaces. On the other hand, the presence of a fluid interface generates in addition interface-mediated (capillary) interactions which are either induced by nonspherical colloid shapes or by the "direct" interactions.     

Visible absorption spectra of metal-catecholate and metal-tironate complexes

Interactions between metals and catechol (1,2-dihydroxybenzene) or other ortho-dihydroxy moieties are being found in an increasing number of biological systems with functions ranging from metal ion internalization to biomaterial synthesis. Although metal-catecholate interactions have been studied in the past, we present the first systematic study of an array of these compounds, all prepared under identical conditions. We report the ultraviolet-visible absorption (UV-vis) spectra for catecholate and tironate complexes of the first row transition elements. Generation and identification of these species were accomplished by preparing aqueous solutions with varied ligand:metal ratios and subsequently titrating with base (NaOH). Controlled ligand deprotonation and metal binding resulted in sequential formation of complexes with one, two, and sometimes three catecholate or tironate ligands bound to a metal ion. We prepared the mono-, bis- and tris-catecholates and -tironates of Fe(3+), V(3+), V(4+)and Mn(3+), the mono- and bis-catecholates and -tironates of Cu(2+), Co(2+), Ni(2+), Zn(2+), Cr(2+) and Mn(2+), and several Ti(4+) and Cr(3+) species. The UV-vis spectra of each complex are described, some of which have not been reported previously. These data can now be applied to characterization of biological metal-catecholate systems.     

Split-luciferase complementary assay: applications,recent developments,and future perspectives

Bioluminescent systems are considered as potent reporter systems for bioanalysis since they have specific characteristics, such as relatively high quantum yields and photon emission over a wide range of colors from green to red. Biochemical events are mostly accomplished through large protein machines. These molecular complexes are built from a few to many proteins organized through their interactions. These protein–protein interactions are vital to facilitate the biological activity of cells. The split-luciferase complementation assay makes the study of two or more interacting proteins possible. In this technique, each of the two domains of luciferase is attached to each partner of two interacting proteins. On interaction of those proteins, luciferase fragments are placed close to each other and form a complemented luciferase, which produces a luminescent signal. Split luciferase is an effective tool for assaying biochemical metabolites, where a domain or an intact protein is inserted into an internally fragmented luciferase, resulting in ligand binding, which causes a change in the emitted signals. We review the various applications of this novel luminescent biosensor in studying protein–protein interactions and assaying metabolites involved in analytical biochemistry, cell communication and cell signaling, molecular biology, and the fate of the whole cell, and show that luciferase-based biosensors are powerful tools that can be applied for diagnostic and therapeutic purposes.     

Magnetic solid phase extraction as a valuable tool for elemental speciation analysis

Despite the increasing number of articles on trace elemental speciation with magnetic solid phase extraction (MSPE), there are no dedicated reviews that cover the group of elements with most related literature, and hence the need for this one. This article provides a comprehensive review of the relevant literature related to Cr, Hg, As, Se, and other metals and metalloids with a special focus on the sorbents, species determined, interactions involved between them and applications, mainly to environmental, food and biological samples. Moreover, this review covers the analysis of metallic nanoparticles (NPs) and the ions that are generated from them as a new facet of speciation. The analytical performance of the methods is addressed from a presentative and critical point of view and, finally, future trends and the related challenges are shown.     

Microfluidics in the “Open Space” for Performing Localized Chemistry on Biological Interfaces

Local interactions between (bio)chemicals and biological interfaces play an important role in fields ranging from surface patterning to cell toxicology. These interactions can be studied using microfluidic systems that operate in the “open space”, that is, without the need for the sealed channels and chambers commonly used in microfluidics. This emerging class of techniques localizes chemical reactions on biological interfaces or specimens without imposing significant “constraints” on samples, such as encapsulation, pre‐processing steps, or the need for scaffolds. They therefore provide new opportunities for handling, analyzing, and interacting with biological samples. The motivation for performing localized chemistry is discussed, as are the requirements imposed on localization techniques. Three classes of microfluidic systems operating in the open space, based on microelectrochemistry, multiphase transport, and hydrodynamic flow confinement of liquids are presented.