Resolution and structural transitions of elongated states of ubiquitin

Electrospray ionization, combined with two-dimensional ion mobility spectrometry and mass spectrometry, is used to produce, select, and activate distributions of elongated ions, [M + 11H]11+ to [M + 13H]13+, of ubiquitin. The analysis makes it possible to examine state-to-state transitions for structural types, and transition diagrams associated with the efficiencies of structural changes are presented. The +11 and +12 charge states can form four resolvable states while only one state is formed for [M + 13H]13+. Some conformations, which appear to belong to the same family based on mobility analysis of different charge states, undergo similar transitions, others do not. Activation of ions that exist in low-abundance conformations, having mobilities that fall in between sharp peaks associated with higher abundances species, shows that the low-abundance forms undergo efficient (approximately 90 to 100%) conversion into states associated with well-defined peaks. This efficiency is significantly higher than the approximately 10 to 60% efficiency of transitions of structures associated with well-defined peaks. The formation of sharp features from a range of low-intensity species with different cross sections indicates that large regions of conformation space must be unfavorable or inaccessible in the gas phase. These results are compared with several previous IMS measurements of this system as well as information about gas-phase structure provided by other techniques.     

Evidence for many resolvable structures within conformation types of electrosprayed ubiquitin ions

A new two-dimensional ion mobility spectrometry approach combined with mass spectrometry has been used to examine ubiquitin ions in the gas phase. In this approach ions are separated in an initial drift tube into conformation types (defined by their collision cross sections) and then a gate is used to introduce a narrow distribution of mobility-separated ions into a second drift tube for subsequent separation. The results show that upon selection a narrow peak shape is retained through the second drift tube. This requires that at 300 K the selected distribution does not interconvert substantially within the broader range of structures associated with the conformation type within the approximately 10-20 ms time scale of these experiments. For the [M + 7H]7+ ion, it appears that many ( approximately 5-10) narrow selections can be made across each of the compact, partially-folded, and elongated conformer types, defined previously (Int. J. Mass Spectrom. 1999, 187, 37-47).     

An inexpensive, fast, and reliable method for vacuum extraction of soil and plant water for stable isotope analyses by mass spectrometry

The stable isotopes of water (hydrogen and oxygen isotopes) are of utmost interest in ecology and the geosciences. In many cases water has to be extracted directly from a matrix such as soil or plant tissue before isotopes can be analyzed by mass spectrometry. Currently, the most widely used technique for water is cryogenic vacuum extraction. We present a simple and inexpensive modification of this method and document tests conducted with soils of various grain size and tree core replicates taken on four occasions during 2010. The accuracies for sandy soils are between 0.4‰ and 3‰ over a range of 21‰ and 165‰ for δ¹⁸O and δ²H, respectively. Spiking tests with water of known isotope composition were conducted with soil and tree core samples; they indicate reliable precision after an extraction time of 15 min for sandy soils. For clayey soils and tree cores, the deviations were up to 0.63‰ and 4.7‰ for δ¹⁸O and δ²H, respectively. This indicates either that the extraction time should be extended or that mechanisms different from Rayleigh fractionation play a role. The modified protocol allows a fast and reliable extraction of large numbers of water samples from soil and plant material in preparation for stable isotope analyses. Copyright © 2011 John Wiley & Sons, Ltd.     

Hydrogeochemical and isotope study of perched aquifers in the Cuvelai-Etosha Basin,Namibia

A hydrogeochemical and stable isotope study (²H and ¹⁸O) was carried out in the Cuvelai-Etosha Basin in order to characterize available groundwater and to identify possible recharge mechanisms for the perched aquifers. Data were collected during seven field campaigns between 2013 and 2015 from a total of 24 shallow and deep groundwater hand-dug wells. In the investigated groundwaters, hydrogencarbonate is the dominating anion in both well types, whereas cations vary between calcium and magnesium in deep wells, and sodium and potassium in shallow wells. Groundwater chemistry is controlled by dissolution of carbonate minerals, silicate weathering and ion exchange. Stable isotopic composition suggests that deep groundwater is recharged by high-intensity/large rainfall events, whereas the shallow wells can even be recharged by less-intense/small rainfall events. Water in deep wells reflect a mixture of water influenced by evaporation during or before infiltration and water that infiltrated through fast preferential pathways, whereas shallow wells are strongly influenced by evaporation. The findings of this research contribute to improve the understanding of hydrogeochemistry, recharge paths and temporal variations of perched aquifers.     

Evidence for spontaneous resolution of icosahedral proline

Here we show experimental evidence for the spontaneous chiral resolution of icosahedral [12Pro+H]+ cluster ion. Molecular simulations reveal that the icosahedron consists of 12 equally spaced prolines where the rigid pyrrolidine ring of each monomer is sticking out of the closed cage. The tightly packed chiral cage traps a single proton in the center cavity. On the other hand, racemic [12Pro+H]+ cluster size exhibits a prismatic structure that can easily incorporate and lose proline monomeric unit sequentially, thus easily forming other geometries. Mechanisms which account for these observations are discussed.     

Estimation of groundwater recharge via deuterium labelling in the semi-arid Cuvelai-Etosha Basin,Namibia

The stable water isotope deuterium (²H) was applied as an artificial tracer (²H₂O) in order to estimate groundwater recharge through the unsaturated zone and describe soil water movement in a semi-arid region of northern central Namibia. A particular focus of this study was to assess the spatiotemporal persistence of the tracer when applied in the field on a small scale under extreme climatic conditions and to propose a method to obtain estimates of recharge in data-scarce regions. At two natural sites that differ in vegetation cover, soil and geology, 500?ml of a 70?% ²H₂O solution was irrigated onto water saturated plots. The displacement of the ²H peak was analyzed 1 and 10 days after an artificial rain event of 20 mm as well as after the rainy season. Results show that it is possible to apply the peak displacement method for the estimation of groundwater recharge rates in semi-arid environments via deuterium labelling. Potential recharge for the rainy season 2013/2014 was calculated as 45 mm a^?1 at 5.6 m depth and 40 mm a^?1 at 0.9 m depth at the two studied sites, respectively. Under saturated conditions, the artificial rain events moved 2.1 and 0.5 m downwards, respectively. The tracer at the deep sand site (site 1) was found after the rainy season at 5.6 m depth, corresponding to a displacement of 3.2 m. This equals in an average travel velocity of 2.8 cm d^?1 during the rainy season at the first site. At the second location, the tracer peak was discovered at 0.9 m depth; displacement was found to be only 0.4 m equalling an average movement of 0.2 cm d^?1 through the unsaturated zone due to an underlying calcrete formation. Tracer recovery after one rainy season was found to be as low as 3.6?% at site 1 and 1.9?% at site 2. With an in situ measuring technique, a three-dimensional distribution of ²H after the rainy season could be measured and visualized. This study comprises the first application of the peak displacement method using a deuterium labelling technique for the estimation of groundwater recharge in semi-arid regions. Deuterium proved to be a suitable tracer for studies within the soil–vegetation–atmosphere interface. The results of this study are relevant for the design of labelling experiments in the unsaturated zone of dry areas using ²H₂O as a tracer and obtaining estimations of groundwater recharge on a local scale. The presented methodology is particularly beneficial in data-scarce environments, where recharge pathways and mechanisms are poorly understood.     

Stable isotope signatures of meteoric water in the Cuvelai-Etosha Basin,Namibia: Seasonal characteristics,trends and relations to southern African patterns

ABSTRACT

The study area is the Namibian part of the Cuvelai-Etosha Basin (CEB), located in central northern Namibia. The CEB is home to 40 % of Namibia’s population, and most of the people live in rural areas. These people depend on both surface and groundwater resources which are limited in this dryland (mean annual rainfall ranging from 250 to 550?mm/a). The isotopic signatures of δ¹⁸O and δ²H from water samples (n?=?61) collected over a course of 9 years from various research projects and existing (but mainly unpublished) data of meteoric water of the CEB (10 sites) were evaluated and local meteoric water lines (LMWLs) developed. Further, the data is discussed in the context of seasonal characteristics and trends and compared to available data from the Global Network of Isotopes in Precipitation (GNIP) for the southern African region. Our results extend the portfolio of previously published LMWLs for southern Africa and provide a more precise baseline for any isotope-based study in that region. The slope of the LMWL from the GNIP stations correlates with latitude. This correlation cannot be found within the CEB. The dominant control on the isotopic signature of the CEB of precipitation is seasonal.     

Analysis of chloroquine and metabolites directly from whole‐body animal tissue sections by liquid extraction surface analysis (LESA) and tandem mass spectrometry

The rapid and direct analysis of the amount and spatial distribution of exogenous chloroquine (CHQ) and CHQ metabolites from tissue sections by liquid extraction surface sampling analysis coupled with tandem mass spectrometry (LESA‐MS/MS) was demonstrated. LESA‐MS/MS results compared well with previously published CHQ quantification data collected by organ excision, extraction and fluorescent detection. The ability to directly sample and analyze spatially resolved exogenous molecules from tissue sections with minimal sample preparation and analytical method development has the potential to facilitate the assessment of target tissue penetration of pharmaceutical compounds, to establish pharmacokinetic/pharmacodynamic relationships, and to complement established pharmacokinetic methods used in the drug discovery process during tissue distribution assessment. Copyright © 2012 John Wiley & Sons, Ltd.     

Imaging cytoplasmic cAMP in mouse brainstem neurons

Background  

cAMP is an ubiquitous second messenger mediating various neuronal functions, often as a consequence of increased intracellular Ca²⁺ levels. While imaging of calcium is commonly used in neuroscience applications, probing for cAMP levels has not yet been performed in living vertebrate neuronal tissue before.