Magnetische Multischalenpartikel für den Transport von siRNA

Die vorliegende Arbeit befasst sich mit der Entwicklung eines nichtviralen, effizienten Transfektionsmittels mit einer Kern-Schale-Struktur in der Größenordnung bis 100 nm. Dafür werden magnetische, negativ geladene Eisenoxid-Nanopartikel mittels Thermolyse mit einem Durchmesser von 17 nm synthetisiert und in Wasser überführt. Diese Nanopartikel bilden den Kern des Erbgut-Trägers und werden mittels Layer-by-Layer –Verfahren (LbL) mit geladenen Polymeren, den bioabbaubaren Makromolekülen Poly-L-Lysin und Heparin, beschichtet. Dafür wird zunächst eine geeignete Apparatur aufgebaut. Diese wird zur Herstellung von Kern-Schale-Strukturen mit fünf Polyelektrolytschichten verwendet und liefert Partikel mit einem hydrodynamischen Durchmesser von 58 nm, die bei Abwesenheit von niedermolekularem Salz aggregatfrei sind. Das System wird gegen Salz stabilisiert, indem die letzte Poly-L-Lysin-Schicht mit Polyethylenglycol modifiziert wird. Die so entstandenen Multischalenpartikel zeigen weder im PBS-Puffer noch in humanem Serum Aggregation. Mittels winkelabhängiger dynamischer Lichtstreuung wird die Aggregatbildung kontrolliert, während ζ-Potential-Messungen die Kontrolle der Oberflächenladung erlauben.rnDa siRNA auf Grund ihres negativ geladenen Phosphat-Rückgrats ebenfalls ein Polyelektrolyt ist, wird sie aggregatfrei auf die positiv geladenen PLL-Nanopartikel aufgetragen. Die eingesetzte siRNA ist farbstoffmarkiert, um eine Detektion in vitro zu ermöglichen. Jedoch sind die entstandenen Komplexe mittels Fluoreszenzkorrelations-spektroskopie (FCS) nicht nachweisbar. Auch die Fluoreszenzmarkierung der PEGylierten Außenschale mittels kupferfreier Click-Chemie ist in der FCS nicht sichtbar, sodass eine Fluoreszenzauslöschung der Farbstoffe in den Multischalenpartikeln vermutet wird.rn

Thermoelectric performance of p-type TiCoSb half-Heusler compounds - Intrinsic phase separation and charge carrier concentration optimization as key to high efficiency

The world's rising demand of energy turns the development of sustainable and more efficient technologies for energy production and storage into an inevitable task. Thermoelectric generators, composed of pairs of n-type and p-type semiconducting materials, di¬rectly transform waste heat into useful electricity. The efficiency of a thermoelectric mate¬rial depends on its electronic and lattice properties, summarized in its figure of merit ZT. Desirable are high electrical conductivity and Seebeck coefficients, and low thermal con¬ductivity. Half-Heusler materials are very promising candidates for thermoelectric applications in the medium¬ temperature range such as in industrial and automotive waste heat recovery. The advantage of Heusler compounds are excellent electronic properties and high thermal and mechanical stability, as well as their low toxicity and elemental abundance. Thus, the main obstacle to further enhance their thermoelectric performance is their relatively high thermal conductivity.rn rnIn this work, the thermoelectric properties of the p-type material (Ti/Zr/Hf)CoSb1-xSnx were optimized in a multistep process. The concept of an intrinsic phase separation has recently become a focus of research in the compatible n-type (Ti/Zr/Hf)NiSn system to achieve low thermal conductivities and boost the TE performance. This concept is successfully transferred to the TiCoSb system. The phase separation approach can form a significant alternative to the previous nanostructuring approach via ball milling and hot pressing, saving pro¬cessing time, energy consumption and increasing the thermoelectric efficiency. A fundamental concept to tune the performance of thermoelectric materials is charge carrier concentration optimization. The optimum carrier concentration is reached with a substitution level for Sn of x = 0.15, enhancing the ZT about 40% compared to previous state-of-the-art samples with x = 0.2. The TE performance can be enhanced further by a fine-tuning of the Ti-to-Hf ratio. A correlation of the microstructure and the thermoelectric properties is observed and a record figure of merit ZT = 1.2 at 710°C was reached with the composition Ti0.25Hf0.75CoSb0.85Sn0.15.rnTowards application, the long term stability of the material under actual conditions of operation are an important issue. The impact of such a heat treatment on the structural and thermoelectric properties is investigated. Particularly, the best and most reliable performance is achieved in Ti0.5Hf0.5CoSb0.85Sn0.15, which reached a maximum ZT of 1.1 at 700°C. The intrinsic phase separation and resulting microstructure is stable even after 500 heating and cooling cycles.

Laboratory-based ROTEM(®) analysis: implementing pneumatic tube transport and real-time graphic transmission

ROTEM(®) is considered a helpful point-of-care device to monitor blood coagulation. Centrally performed analysis is desirable but rapid transport of blood samples and real-time transmission of graphic results are an important prerequisite. The effect of sample transport through a pneumatic tube system on ROTEM(®) results is unknown. The aims of the present work were (i) to determine the influence of blood sample transport through a pneumatic tube system on ROTEM(®) parameters compared to manual transportation, and (ii) to verify whether graphic results can be transmitted on line via virtual network computing using local area network to the physician in charge of the patient.

Transport model of the human Na+-coupled L-ascorbic acid (vitamin C) transporter SVCT1

Vitamin C (L-ascorbic acid) is an essential micronutrient that serves as an antioxidant and as a cofactor in many enzymatic reactions. Intestinal absorption and renal reabsorption of the vitamin is mediated by the epithelial apical L-ascorbic acid cotransporter SVCT1 (SLC23A1). We explored the molecular mechanisms of SVCT1-mediated L-ascorbic acid transport using radiotracer and voltage-clamp techniques in RNA-injected Xenopus oocytes. L-ascorbic acid transport was saturable (K(0.5) approximately 70 microM), temperature dependent (Q(10) approximately 5), and energized by the Na(+) electrochemical potential gradient. We obtained a Na(+)-L-ascorbic acid coupling ratio of 2:1 from simultaneous measurement of currents and fluxes. L-ascorbic acid and Na(+) saturation kinetics as a function of cosubstrate concentrations revealed a simultaneous transport mechanism in which binding is ordered Na(+), L-ascorbic acid, Na(+). In the absence of L-ascorbic acid, SVCT1 mediated pre-steady-state currents that decayed with time constants 3-15 ms. Transients were described by single Boltzmann distributions. At 100 mM Na(+), maximal charge translocation (Q(max)) was approximately 25 nC, around a midpoint (V(0.5)) at -9 mV, and with apparent valence approximately -1. Q(max) was conserved upon progressive removal of Na(+), whereas V(0.5) shifted to more hyperpolarized potentials. Model simulation predicted that the pre-steady-state current predominantly results from an ion-well effect on binding of the first Na(+) partway within the membrane electric field. We present a transport model for SVCT1 that will provide a framework for investigating the impact of specific mutations and polymorphisms in SLC23A1 and help us better understand the contribution of SVCT1 to vitamin C metabolism in health and disease.

Analysis of transport properties of mechanically alloyed [Pb1-xSnxTe]

The work described in this thesis had two objectives. The first objective was to develop a physically based computational model that could be used to predict the electronic conductivity, Seebeck coefficient, and thermal conductivity of Pb1-xSnxTe alloys over the 400 K to 700 K temperature as a function of Sn content and doping level. The second objective was to determine how the secondary phase inclusions observed in Pb1-xSnxTe alloys made by consolidating mechanically alloyed elemental powders impact the ability of the material to harvest waste heat and generate electricity in the 400 K to 700 K temperature range. The motivation for this work was that though the promise of this alloy as an unusually efficient thermoelectric power generator material in the 400 K to 700 K range had been demonstrated in the literature, methods to reproducibly control and subsequently optimize the materials thermoelectric figure of merit remain elusive. Mechanical alloying, though not typically used to fabricate these alloys, is a potential method for cost-effectively engineering these properties. Given that there are deviations from crystalline perfection in mechanically alloyed material such as secondary phase inclusions, the question arises as to whether these defects are detrimental to thermoelectric function or alternatively, whether they enhance thermoelectric function of the alloy. The hypothesis formed at the onset of this work was that the small secondary phase SnO2 inclusions observed to be present in the mechanically alloyed Pb1-xSnxTe would increase the thermoelectric figure of merit of the material over the temperature range of interest. It was proposed that the increase in the figure of merit would arise because the inclusions in the material would not reduce the electrical conductivity to as great an extent as the thermal conductivity. If this were to be true, then the experimentally measured electronic conductivity in mechanically alloyed Pb1-xSnxTe alloys that have these inclusions would not be less than that expected in alloys without these inclusions while the portion of the thermal conductivity that is not due to charge carriers (the lattice thermal conductivity) would be less than what would be expected from alloys that do not have these inclusions. Furthermore, it would be possible to approximate the observed changes in the electrical and thermal transport properties using existing physical models for the scattering of electrons and phonons by small inclusions. The approach taken to investigate this hypothesis was to first experimentally characterize the mobile carrier concentration at room temperature along with the extent and type of secondary phase inclusions present in a series of three mechanically alloyed Pb1-xSnxTe alloys with different Sn content. Second, the physically based computational model was developed. This model was used to determine what the electronic conductivity, Seebeck coefficient, total thermal conductivity, and the portion of the thermal conductivity not due to mobile charge carriers would be in these particular Pb1-xSnxTe alloys if there were to be no secondary phase inclusions. Third, the electronic conductivity, Seebeck coefficient and total thermal conductivity was experimentally measured for these three alloys with inclusions present at elevated temperatures. The model predictions for electrical conductivity and Seebeck coefficient were directly compared to the experimental elevated temperature electrical transport measurements. The computational model was then used to extract the lattice thermal conductivity from the experimentally measured total thermal conductivity. This lattice thermal conductivity was then compared to what would be expected from the alloys in the absence of secondary phase inclusions. Secondary phase inclusions were determined by X-ray diffraction analysis to be present in all three alloys to a varying extent. The inclusions were found not to significantly degrade electrical conductivity at temperatures above ~ 400 K in these alloys, though they do dramatically impact electronic mobility at room temperature. It is shown that, at temperatures above ~ 400 K, electrons are scattered predominantly by optical and acoustical phonons rather than by an alloy scattering mechanism or the inclusions. The experimental electrical conductivity and Seebeck coefficient data at elevated temperatures were found to be within ~ 10 % of what would be expected for material without inclusions. The inclusions were not found to reduce the lattice thermal conductivity at elevated temperatures. The experimentally measured thermal conductivity data was found to be consistent with the lattice thermal conductivity that would arise due to two scattering processes: Phonon phonon scattering (Umklapp scattering) and the scattering of phonons by the disorder induced by the formation of a PbTe-SnTe solid solution (alloy scattering). As opposed to the case in electrical transport, the alloy scattering mechanism in thermal transport is shown to be a significant contributor to the total thermal resistance. An estimation of the extent to which the mean free time between phonon scattering events would be reduced due to the presence of the inclusions is consistent with the above analysis of the experimental data. The first important result of this work was the development of an experimentally validated, physically based computational model that can be used to predict the electronic conductivity, Seebeck coefficient, and thermal conductivity of Pb1-xSnxTe alloys over the 400 K to 700 K temperature as a function of Sn content and doping level. This model will be critical in future work as a tool to first determine what the highest thermoelectric figure of merit one can expect from this alloy system at a given temperature and, second, as a tool to determine the optimum Sn content and doping level to achieve this figure of merit. The second important result of this work is the determination that the secondary phase inclusions that were observed to be present in the Pb1-xSnxTe made by mechanical alloying do not keep the material from having the same electrical and thermal transport that would be expected from “perfect" single crystal material at elevated temperatures. The analytical approach described in this work will be critical in future investigations to predict how changing the size, type, and volume fraction of secondary phase inclusions can be used to impact thermal and electrical transport in this materials system.

¿Impact from road transport of radioactive wastes in Spain?

Transports of radioactive wastes in Spain are becoming issues of renewed interest, due to the increased mobility of these materials which can be expected after the building and operation of the planned central repository for this country in a near future. Such types of residues will be mainly of the medium and high activity classes and have raised concerns on the safety of the operations, the radiological protection of the individuals, the compliance with the legal regulations and their environmental consequences of all kind. In this study, relevant information for the assessment of radiological risk of road transport were taken into account, as the sources and destination of the radioactive transports, the amount of traveling to be done, the preferred routes and populations affected, the characterization of the residues and containers, their corresponding testing, etc. These data were supplied by different organizations fully related with these activities, like the nuclear power stations, the companies in charge of radioactive transports, the enterprises for inspection and control of the activities, etc., as well as the government institutions which are responsible for the selection and location of the storage facility and other decisions on the nuclear policies of the country. Thus, we have developed a program for computing the data in such a form that by entering the radiation levels at one meter of the transport loads and by choosing a particular displacement, the computer application is capable to calculate the corresponding radiological effects, like the global estimated impact, its relevance to the population in general or on those people living and driving near the main road routes, the doses received by the most exposed individuals (e.g. the workers for loading or driving the vehicle), or the probability of detrimental on the human health. The results of this work could be of help for a better understanding and management of these activities and their related impacts; at the same time that the generated reports of the computer application are considered of particular interest as innovative and complementary information to the current legal documentation, which is basically required for transporting radioactive wastes in the country, according with the international safety rules (like IAEA and ADR).Though main studies are still in progress, as the definite location for the Spanish storage facility has not been decided yet, preliminary results with the existing transports of residues of medium activity indicate that the radiological impact is very low in conventional operations. Nevertheless, the management of these transports is complex and laborious, making it convenient to progress further in the analysis and quantification of this kind of events, which constitutes one of the main objectives of the present study for the radioactive road mobility in Spain.

Modeling impact assessment of transport policies through a multiregional input-output integrated approach

El objetivo de esta investigación es desarrollar una metodología para estimar los potenciales impactos económicos y de transporte generados por la aplicación de políticas en el sector transporte. Los departamentos de transporte y otras instituciones gubernamentales relacionadas se encuentran interesadas en estos análisis debido a que son presentados comúnmente de forma errónea por la insuficiencia de datos o por la falta de metodologías adecuadas. La presente investigación tiene por objeto llenar este vacío haciendo un análisis exhaustivo de las técnicas disponibles que coincidan con ese propósito. Se ha realizado un análisis que ha identificado las diferencias cuando son aplicados para la valoración de los beneficios para el usuario o para otros efectos como aspectos sociales. Como resultado de ello, esta investigación ofrece un enfoque integrado que incluye un modelo Input-Output de múltiples regiones basado en la utilidad aleatoria (RUBMRIO), y un modelo de red de transporte por carretera. Este modelo permite la reproducción con mayor detalle y realismo del transporte de mercancías que por medio de su estructura sectorial identifica los vínculos de las compras y ventas inter-industriales dentro de un país utilizando los servicios del transporte de mercancías. Por esta razón, el modelo integrado es aplicable a diversas políticas de transporte. En efecto, el enfoque se ha aplicado para estudiar los efectos macroeconómicos regionales de la implementación de dos políticas diferentes en el sistema de transporte de mercancías de España, tales como la tarificación basada en la distancia recorrida por vehículo-kilómetro (€/km) aplicada a los vehículos del transporte de mercancías, y para la introducción de vehículos más largos y pesados de mercancías en la red de carreteras de España. El enfoque metodológico se ha evaluado caso por caso teniendo en cuenta una selección de la red de carreteras que unen las capitales de las regiones españolas. También se ha tenido en cuenta una dimensión económica a través de una tabla Input-Output de múltiples regiones (MRIO) y la base de datos de conteo de tráfico existente para realizar la validación del modelo. El enfoque integrado reproduce las condiciones de comercio observadas entre las regiones usando el sistema de transporte de mercancías por carretera, y que permite por comparación con los escenarios de políticas, determinar las contribuciones a los cambios distributivos y generativos. Así pues, el análisis estima los impactos económicos en cualquier región considerando los cambios en el Producto Interno Bruto (PIB) y el empleo. El enfoque identifica los cambios en el sistema de transporte a través de todos los caminos de la red de transporte a través de las medidas de efectividad (MOEs). Los resultados presentados en esta investigación proporcionan evidencia sustancial de que en la evaluación de las políticas de transporte, es necesario establecer un vínculo entre la estructura económica de las regiones y de los servicios de transporte. Los análisis muestran que para la mayoría de las regiones del país, los cambios son evidentes para el PIB y el empleo, ya que el comercio se fomenta o se inhibe. El enfoque muestra cómo el tráfico se desvía en ambas políticas, y también determina detalles de las emisiones de contaminantes en los dos escenarios. Además, las políticas de fijación de precios o de regulación de los sistemas de transporte de mercancías por carretera dirigidas a los productores y consumidores en las regiones promoverán transformaciones regionales afectando todo el país, y esto conduce a conclusiones diferentes. Así mismo, este enfoque integrado podría ser útil para evaluar otras políticas y otros países en todo el mundo. The purpose of this research is to develop a methodological approach aimed at assessing the potential economic and transportation impacts of transport policies. Transportation departments and other related government parties are interested in such analysis because it is commonly misrepresented for the insufficiency of data and suitable methodologies available. This research is directed at filling this gap by making a comprehensive analysis of the available techniques that match with that purpose. The differences when they are applied for the valuation of user benefits or for other impacts as social matters have been identified. As a result, this research presents an integrated approach which includes both a random utility-based multiregional Input-Output model (RUBMRIO), and a road transport network model. This model accounts for freight transport with more detail and realism because its commodity-based structure traces the linkages of inter-industry purchases and sales that use freight services within a given country. For this reason, the integrated model is applicable to various transport policies. In fact, the approach is applied to study the regional macroeconomic effects of implementing two different policies in the freight transport system of Spain, such as a distance-based charge in vehicle-kilometer (€/km) for Heavy Goods Vehicles (HGVs), and the introduction of Longer and Heavier Vehicles (LHVs) in the road network of Spain. The methodological approach has been evaluated on a case by case basis considering a selected road network of highways linking the capitals of the Spanish regions. It has also considered an economic dimension through a Multiregional Input Output Table (MRIO) and the existing traffic count database used in the model validation. The integrated approach replicates observed conditions of trade among regions using road freight transport systems that determine contributions to distributional and generative changes by comparison with policy scenarios. Therefore, the model estimates economic impacts in any given area by considering changes in Gross Domestic Product (GDP), employment (jobs), and in the transportation system across all paths of the transport network considering Measures of effectiveness (MOEs). The results presented in this research provide substantive evidence that in the assessment of transport policies it is necessary to establish a link between the economic structure of regions and the transportation services. The analysis shows that for most regions in the country, GDP and employment changes are noticeable when trade is encouraged or discouraged. This approach shows how traffic is diverted in both policies, and also provides details of the pollutant emissions in both scenarios. Furthermore, policies, such as pricing or regulation of road freight transportation systems, directed to producers and consumers in regions will promote different regional transformations across the country, and this lead to different conclusions. In addition, this integrated approach could be useful to assess other policies and countries worldwide.

Interregional trade changes in Spain caused by the introduction of a road fee charge for heavy goods vehicles

The introduction of a homogeneous road charging system according to the Directive 2011/76/EU for the use of roads is still under development in most European Union (EU) member states. Spain, like other EU members, has been encouraged to introduce a charging system for Heavy Goods Vehicles (HGVs) throughout the country. This nationwide charge has been postponed because there are serious concerns about their advantages from an economic point of view. Within this context, this paper applies an integrated modeling approach to shape elastic trade coefficients among regions by using a random utility based multiregional Input- Output (RUBMRIO) approach and a road transport network model in order to determine regional distributive and substitutive economic effects by simulating the introduction of a distance-based charge (?/km) considering 7,053.8 kilometers of free highways linking the capitals of the Spanish regions. In addition, an in-depth analysis of interregional trade changes is developed to evaluate and characterize the role of the road charging approach in trade relations among regions and across freight intensive economic sectors. For this purpose, differences in trade relations are described and assessed between a base-case or ?do nothing? scenario and a road fee-charge setting scenario. The results show that the specific amount of the charge set for HGVs affect each region differently and to a different extent because in some regions the price of commodities and the Generalized Transport Cost will decrease its competiveness within the country.

Conformational changes couple Na+ and glucose transport

The mechanism by which cotransport proteins couple their substrates across cell membranes is not known. A commonly proposed model is that cotransport results from ligand-induced conformational transitions that change the accessibility of ligand-binding sites from one side of the membrane to the other. To test this model, we have measured the accessibility of covalent probes to a cysteine residue (Q457C) placed in the putative sugar-translocation domain of the Na+/glucose cotransporter (SGLT1). The mutant protein Q457C was able to transport sugar, but transport was abolished after alkylation by methanethiosulfonate reagents. Alkylation blocked sugar translocation but not sugar binding. Accessibility of Q457C to alkylating reagents required external Na+ and was blocked by external sugar and phlorizin. The voltage dependence of accessibility was directly correlated with the presteady–state charge movement of SGLT1. Voltage-jump experiments with rhodamine-6-maleimide-labeled Q457C showed that the time course and level of changes in fluorescence closely followed the presteady–state charge movement. We conclude that conformational changes are responsible for the coupling of Na+ and sugar transport and that Q457 plays a critical role in sugar translocation by SGLT1.

Dynamical principles in biological processes: A model of charge migration in proteins and DNA

The generalized master equations (GMEs) that contain multiple time scales have been derived quantum mechanically. The GME method has then been applied to a model of charge migration in proteins that invokes the hole hopping between local amino acid sites driven by the torsional motions of the floppy backbones. This model is then applied to analyze the experimental results for sequence-dependent long-range hole transport in DNA reported by Meggers et al. [Meggers, E., Michel-Beyerle, M. E., & Giese, B. (1998) J. Am. Chem. Soc. 120, 12950–12955]. The model has also been applied to analyze the experimental results of femtosecond dynamics of DNA-mediated electron transfer reported by Zewail and co-workers [Wan, C., Fiebig, T., Kelley, S. O., Treadway, C. R., Barton, J. K. & Zewail, A. H. (1999) Proc. Natl. Acad. Sci. USA 96, 6014–6019]. The initial events in the dynamics of protein folding have begun to attract attention. The GME obtained in this paper will be applicable to this problem.

Nicotianamine Chelates Both FeIII and FeII. Implications for Metal Transport in Plants1

Nicotianamine (NA) occurs in all plants and chelates metal cations, including FeII, but reportedly not FeIII. However, a comparison of the FeII and ZnII affinity constants of NA and various FeIII-chelating aminocarboxylates suggested that NA should chelate FeIII. High-voltage electrophoresis of the FeNA complex formed in the presence of FeIII showed that the complex had a net charge of 0, consistent with the hexadentate chelation of FeIII. Measurement of the affinity constant for FeIII yielded a value of 1020.6, which is greater than that for the association of NA with FeII (1012.8). However, capillary electrophoresis showed that in the presence of FeII and FeIII, NA preferentially chelates FeII, indicating that the FeIINA complex is kinetically stable under aerobic conditions. Furthermore, Fe complexes of NA are relatively poor Fenton reagents, as measured by their ability to mediate H2O2-dependent oxidation of deoxyribose. This suggests that NA will have an important role in scavenging Fe and protecting the cell from oxidative damage. The pH dependence of metal ion chelation by NA and a typical phytosiderophore, 2′-deoxymugineic acid, indicated that although both have the ability to chelate Fe, when both are present, 2′-deoxymugineic acid dominates the chelation process at acidic pH values, whereas NA dominates at alkaline pH values. The consequences for the role of NA in the long-distance transport of metals in the xylem and phloem are discussed.

Arg-302 facilitates deprotonation of Glu-325 in the transport mechanism of the lactose permease from Escherichia coli

A mechanistic model for lactose/H+ symport via the lactose permease of Escherichia coli proposed recently indicates that the permease must be protonated to bind ligand with high affinity. Moreover, in the ground state, the symported H+ is shared between His-322 (helix X) and Glu-269 (helix VIII), whereas Glu-325 (helix X) is charge-paired with Arg-302 (helix IX). Substrate binding at the outer surface induces a conformational change that leads to transfer of the H+ to Glu-325 and reorientation of the binding site to the inner surface. After release of the substrate, Glu-325 is deprotonated on the inside because of rejuxtapositioning with Arg-302. To test the role of Arg-302 in the mechanism, the catalytic properties of mutants Arg-302→Ala and Arg-302→Ser were studied. Both mutants are severely defective in active lactose transport, as well as in efflux or influx down a concentration gradient, translocation modes that involve net H+ movement. In marked contrast, the mutants catalyze equilibrium exchange of lactose and bind ligand with high affinity. These characteristics are remarkably analogous to those of permease mutants with neutral replacements for Glu-325, a residue that plays a direct role in H+ translocation. These observations lend strong support for the argument that Arg-302 interacts with Glu-325 to facilitate deprotonation of the carboxylic acid during turnover.

Characterization of complex mineral assemblages: Implications for contaminant transport and environmental remediation

Surface reactive phases of soils and aquifers, comprised of phyllosilicate and metal oxohydroxide minerals along with humic substances, play a critical role in the regulation of contaminant fate and transport. Much of our knowledge concerning contaminant-mineral interactions at the molecular level, however, is derived from extensive experimentation on model mineral systems. Although these investigations have provided a foundation for understanding reactive surface functional groups on individual mineral phases, the information cannot be readily extrapolated to complex mineral assemblages in natural systems. Recent studies have elucidated the role of less abundant mineral and organic substrates as important surface chemical modifiers and have demonstrated complex coupling of reactivity between permanent-charge phyllosilicates and variable-charge Fe-oxohydroxide phases. Surface chemical modifiers were observed to control colloid generation and transport processes in surface and subsurface environments as well as the transport of solutes and ionic tracers. The surface charging mechanisms operative in the complex mineral assemblages cannot be predicted based on bulk mineralogy or by considering surface reactivity of less abundant mineral phases based on results from model systems. The fragile nature of mineral assemblages isolated from natural systems requires novel techniques and experimental approaches for investigating their surface chemistry and reactivity free of artifacts. A complete understanding of the surface chemistry of complex mineral assemblages is prerequisite to accurately assessing environmental and human health risks of contaminants or in designing environmentally sound, cost-effective chemical and biological remediation strategies.

The Regulation of Photosynthetic Electron Transport during Nutrient Deprivation in Chlamydomonas reinhardtii1

The light-saturated rate of photosynthetic O2 evolution in Chlamydomonas reinhardtii declined by approximately 75% on a per-cell basis after 4 d of P starvation or 1 d of S starvation. Quantitation of the partial reactions of photosynthetic electron transport demonstrated that the light-saturated rate of photosystem (PS) I activity was unaffected by P or S limitation, whereas light-saturated PSII activity was reduced by more than 50%. This decline in PSII activity correlated with a decline in both the maximal quantum efficiency of PSII and the accumulation of the secondary quinone electron acceptor of PSII nonreducing centers (PSII centers capable of performing a charge separation but unable to reduce the plastoquinone pool). In addition to a decline in the light-saturated rate of O2 evolution, there was reduced efficiency of excitation energy transfer to the reaction centers of PSII (because of dissipation of absorbed light energy as heat and because of a transition to state 2). These findings establish a common suite of alterations in photosynthetic electron transport that results in decreased linear electron flow when C. reinhardtii is limited for either P or S. It was interesting that the decline in the maximum quantum efficiency of PSII and the accumulation of the secondary quinone electron acceptor of PSII nonreducing centers were regulated specifically during S-limited growth by the SacI gene product, which was previously shown to be critical for the acclimation of C. reinhardtii to S limitation (J.P. Davies, F.H. Yildiz, and A.R. Grossman [1996] EMBO J 15: 2150–2159).

Electrical currents associated with nucleotide transport by the reconstituted mitochondrial ADP/ATP carrier.

The electrophoretic export of ATP against the import of ADP in mitochondria bridges the intra- versus extramitochondrial ATP potential gap. Here we report that the electrical nature of the ADP/ATP exchange by the mitochondrial ADP/ATP carrier (AAC) can be directly studied by measuring the electrical currents via capacitive coupling of AAC-containing vesicles on a planar lipid membrane. The currents were induced by the rapid liberation of ATP or ADP with UV flash photolysis from caged nucleotides. Six different transport modes of the AAC were studied: heteroexchange with either ADP or ATP inside the vesicles, initiated by photolysis of caged ATP or ADP; homoexchange with ADPex/ADPin or ATPex/ATPin; and caged ADP or ATP with unloaded vesicles. The heteroexchange produced the largest currents with the longest duration in line with the electrical charge difference ATP4- versus ADP3-. Surprisingly, also in the homoexchange and with unloaded vesicles, small currents were measured with shorter duration. In all three modes with caged ATP, a negative charge moved into the vesicles and with caged ADP it moved out of the vesicles. All currents were completely inhibited by a mixture of the inhibitors of the AAC, carboxyatractyloside and hongkrekate, which proves that the currents are exclusively due to AAC function. The observed charge movements in the heteroexchange system agree with the prediction from transport studies in mitochondria and reconstituted vesicles. The unexpected charge movements in the homoexchange or unloaded systems are interpreted to reveal transmembrane rearrangements of charged sites in the AAC when occupied with ADP or ATP. The results also indicate that not only ATP4- but also ADP3- contribute, albeit in opposite direction, to the electrical nature of the ADP/ATP exchange, which is at variance with former conclusions from biochemical transport studies. These measurements open up new avenues of studying the electrical interactions of ADP and ATP with the AAC.