Human hair absorbs numerous biomolecules from the body during its growth. This can act as a fingerprint to determine substance intake of an individual, which can be useful in forensic studies. The ...cocaine concentration profile along the growth axis of hair indicates the time evolution of the metabolic incorporation of cocaine usage. It could be either assessed by chemical extraction and further analysis of hair bundels, or by direct single hair fibre analysis with mass spectroscopy imaging (MSI). Within this work, we analyzed the cocaine distribution in individual hair samples using MeV-SIMS. Unlike conventional surface analysis methods, we demonstrate high yields of nonfragmented molecular ions from the surface of biological materials, resulting in high chemical sensitivity and non-destructive characterisation. Hair samples were prepared by longitudinally cutting along the axis of growth, leaving half-cylindrical shape to access the interior structure of the hair by the probing ion beam, and attached to the silicon wafer. A focused 5.8 MeV 35Cl6+ beam was scanned across the intact, chemically pristine hair structure. A non-fragmented protonated M+ H+ cocaine molecular peak at m/z = 304 was detected and localized along the cross-section of the hair. Its intensity exhibits strong fluctuations along the direction of the hair's growth, with pronounced peaks as narrow as 50 micrometres, corresponding to a metabolic incorporation time of approx. three hours.
Gomphrena claussenii is a recently described zinc (Zn)- and cadmium (Cd)-hypertolerant Amaranthaceae species displaying a metal bioindicator Zn/Cd accumulation response. We investigated the Zn and Cd ...distribution in stem and leaf tissues of G. claussenii at the cellular level, and determined metabolite profiles to investigate metabolite involvement in Zn and Cd sequestration.
Gomphrena claussenii plants exposed to high Zn and Cd supply were analysed by scanning electron microscopy with energy-dispersive X-ray (SEM-EDX) and micro-proton-induced X-ray emission (micro-PIXE). In addition, gas chromatography-time of flight-mass spectrometry (GC-TOF-MS) was used to determine metabolite profiles on high Zn and Cd exposure.
Stem and leaf tissues of G. claussenii plants exposed to control and high Cd conditions showed the abundant presence of calcium oxalate (CaOx) crystals, but on high Zn exposure, their abundance was strongly reduced. Ca and Cd co-localized to the CaOx crystals in Cd-exposed plants. Citrate, malate and oxalate levels were all higher in shoot tissues of metal-exposed plants, with oxalate levels induced 2.6-fold on Zn exposure and 6.4-fold on Cd exposure.
Sequestration of Cd in vacuolar CaOx crystals of G. claussenii is found to be a novel mechanism to deal with Cd accumulation and tolerance.
Jožef Stefan Institute recently commissioned a high brightness H− ion beam injection system for its existing tandem accelerator facility. Custom developed by High Voltage Engineering Europa, the ...multicusp ion source has been tuned to deliver at the entrance of the Tandetron™ accelerator H− ion beams with a measured brightness of 17.1Am−2rad−2eV−1at 170μA, equivalent to an energy normalized beam emittance of 0.767πmmmradMeV1/2. Upgrading the accelerator facility with the new injection system provides two main advantages. First, the high brightness of the new ion source enables the reduction of object slit aperture and the reduction of acceptance angle at the nuclear microprobe, resulting in a reduced beam size at selected beam intensity, which significantly improves the probe resolution for micro-PIXE applications. Secondly, the upgrade strongly enhances the accelerator up-time since H and He beams are produced by independent ion sources, introducing a constant availability of 3He beam for fusion-related research with NRA.
The ion beam particle losses and ion beam emittance growth imply that the aforementioned beam brightness is reduced by transport through the ion optical system. To obtain quantitative information on the available brightness at the high-energy side of the accelerator, the proton beam brightness is determined in the nuclear microprobe beamline. Based on the experience obtained during the first months of operation for micro-PIXE applications, further necessary steps are indicated to obtain optimal coupling of the new ion source with the accelerator to increase the normalized high-energy proton beam brightness at the JSI microprobe, currently at 14Am−2rad−2eV−1, with the output current at 18% of its available maximum.
Cadmium and zinc share many similar physiochemical properties, but their compartmentation, complexation and impact on other mineral element distribution in plant tissues may drastically differ. In ...this study, we address the impact of 10 μm Cd or 50 μm Zn treatments on ion distribution in leaves of a metallicolous population of the non‐hyperaccumulating species Zygophyllum fabago at tissue and cell level, and the consequences on the plant response through a combined physiological, proteomic and metabolite approach. Micro‐proton‐induced X‐ray emission and laser ablation inductively coupled mass spectrometry analyses indicated hot spots of Cd concentrations in the vicinity of vascular bundles in response to Cd treatment, essentially bound to S‐containing compounds as revealed by extended X‐ray absorption fine structure and non‐protein thiol compounds analyses. A preferential accumulation of Zn occurred in vascular bundle and spongy mesophyll in response to Zn treatment, and was mainly bound to O/N‐ligands. Leaf proteomics and physiological status evidenced a protection of photosynthetically active tissues and the maintenance of cell turgor through specific distribution and complexation of toxic ions, reallocation of some essential elements, synthesis of proteins involved in photosynthetic apparatus or C‐metabolism, and metabolite synthesis with some specificities regarding the considered heavy metal treatment.
Iron insufficiency is a worldwide problem in human diets. In cereals like wheat, the bran layer of the grains is an important source of iron. However, the dietary availability of iron in wheat flour ...is limited due to the loss of the iron-rich bran during milling and processing and the presence of anti-nutrients like phytic acid that keep iron strongly chelated in the grain. The present study investigated the localization of iron and phosphorus in grain tissues of wheat genotypes with contrasting grain iron content using synchrotron-based micro-X-ray fluorescence (micro-XRF) and micro-proton-induced X-ray emission (micro-PIXE). X-ray absorption near-edge spectroscopy (XANES) was employed to determine the proportion of divalent and trivalent forms of Fe in the grains. It revealed the abundance of oxygen, phosphorus, and sulphur in the local chemical environment of Fe in grains, as Fe-O-P-R and Fe-O-S-R coordination. Contrasting differences were noticed in tissue-specific relative localization of Fe, P, and S among the different genotypes, suggesting a possible effect of localization pattern on iron bioavailability. The current study reports the shift in iron distribution from maternal to filial tissues of grains during the evolution of wheat from its wild relatives to the present-day cultivated varieties, and thus suggests the value of detailed physical localization studies in varietal improvement programmes for food crops.
MAIN CONCLUSION: Wheat and its related genotypes show distinct distribution patterns for mineral nutrients in maternal and filial tissues in grains. X-ray-based imaging techniques are very ...informative to identify genotypes with contrasting tissue-specific localization of different elements. This can help in the selection of suitable genotypes for nutritional improvement of food grain crops. Understanding mineral localization in cereal grains is important for their nutritional improvement. Spatial distribution of mineral nutrients (Mg, P, S, K, Ca, Fe, Zn, Mn and Cu) was investigated between and within the maternal and filial tissues in grains of two wheat cultivars (Triticum aestivum Cv. WH291 and WL711), a landrace (T. aestivum L. IITR26) and a related wild species Aegilops kotschyi, using micro-proton-induced X-ray emission (µ-PIXE) and micro-X-ray fluorescence (µ-XRF). Aleurone and scutellum were major storage tissues for macro (P, K, Ca and Mg) as well as micro (Fe, Zn, Cu and Mn) nutrients. Distinct elemental distribution patterns were observed in each of the four genotypes. A. kotschyi, the wild relative of wheat and the landrace, T. aestivum L. IITR26, accumulated more Zn and Fe in scutellum and aleurone than the cultivated wheat varieties, WH291 and WL711. The landrace IITR26, accumulated far more S in grains, Mn in scutellum, aleurone and embryo region, Ca and Cu in aleurone and scutellum, and Mg, K and P in scutellum than the other genotypes. Unlike wheat, lower Mn and higher Fe, Cu and Zn concentrations were noticed in the pigment strand of A. kotschyi. Multivariate statistical analysis, performed on mineral distribution in major grain tissues (aleurone, scutellum, endosperm and embryo region) resolved the four genotypes into distinct clusters.
With a model invertebrate animal, we have assessed the fate of magnetic nanoparticles in biologically relevant media, i.e., digestive juices. The toxic potential and the internalization of such ...nanoparticles by nontarget cells were also examined. The aim of this study was to provide experimental evidence on the formation of Co2+, Fe2+, and Fe3+ ions from CoFe2O4 nanoparticles in the digestive juices of a model organism. Standard toxicological parameters were assessed. Cell membrane stability was tested with a modified method for measurement of its quality. Proton-induced X-ray emission and low energy synchrotron radiation X-ray fluorescence were used to study internalization and distribution of Co and Fe. Co2+ ions were found to be more toxic than nanoparticles. We confirmed that Co2+ ions accumulate in the hepatopancreas, but Fe n+ ions or CoFe2O4 nanoparticles are not retained in vivo. A model biological system with a terrestrial isopod is suited to studies of the potential dissolution of ions and other products from metal-containing nanoparticles in biologically complex media.
The different buckwheat species are gluten-free pseudocereals, and they are amongst the most important alternative foodstuffs as they have an excellent nutrient profile. The grain of tartary ...buckwheat (Fagopyrum tataricum Gaertn.) is known to be rich in antioxidants (especially rutin) and mineral elements. In addition to the spatial distributions of Mg, P, S, K, Ca, Mn, Fe, Cu and Zn at the tissue level that have been determined using micro-proton-induced X-ray emission, we have also assessed the distributions of C, O, Mg, P, Mn, Fe, Cu and Zn at the cellular and sub-cellular levels using synchrotron radiation-based low-energy X-ray fluorescence. The highest concentrations of all of the studied mineral elements were in the cotyledons, except for Ca, which dominated in the pericarp. The relative mineral-element distributions calculated on dry weight basis confirm the observed mineral distribution profiles. In the cotyledons, P clearly partitions to the mesophyll and is mainly ascribed to phytate. In the mesophyll, the P spatial distribution strongly correlates with Mg and, in decreasing order, with Cu>Fe>Mn>Zn. These spatial distributions of the mineral elements, their concentrations and their co-localisation are discussed in relation to other pseudocereal and cereal grain.
•Tissue and cell-specific distributions of mineral elements are presented.•Most mineral elements are highest in cotyledons, with Ca mainly in the pericarp.•P localisation correlates strongly with Mg and Fe, in the mesophyll of cotyledons.•Binding sites other than phytate are indicated for Mn, Cu and Zn in the mesophyll.
•Concentration of Mg, P, Cl, K, Cu and Zn in the hippocampus using X-ray spectroscopy.•Concentration of Mg, P, Cl, K and Cu increased during aversive memory consolidation.•Concentrations of Mg, P, Cl ...and K increased during aversive memory reconsolidation.•Element engaged in memory consolidation may be also participating in reconsolidation.
The concentration and distribution of Mg, P, Cl, K, Cu and Zn in the dorsal hippocampus CA1 region of rat brains were studied during memory consolidation and reconsolidation processes stimulated with inhibitory avoidance (IA) tests. Experimental rats were divided into four groups: i) group not submitted to inhibitory avoidance task (IA-N); ii) group submitted to inhibitory avoidance training session (IA-Y); iii) group submitted to inhibitory avoidance reactivation session but did not step down from the platform (IAR-N); and iv) group submitted to avoidance reactivation session and stepped down from the platform (IAR-Y). Elemental concentration and distribution in the CA1 hippocampus region were obtained through the Particle-Induced X-ray Emission (PIXE) technique. The results indicate that the concentration of Mg, P, Cl, K and Cu increased during memory consolidation. During the memory reconsolidation process, the concentrations of Mg, P, Cl and K increased, while Cu and Zn had no significant changes with respect to their basal condition. These results show that the major part of these elements may be engaged in memory consolidation could be also participating in memory reconsolidation. For all elements, the general trend related to their concentration did not change during reconsolidation regardless the presence of a novelty event, i.e. stepping down from the platform.
Particle induced X-ray emission (PIXE) at microprobe of Jožef Stefan Institute is used to measure two-dimensional quantitative elemental maps of biological tissue. To improve chemical and biological ...understanding of the processes in vivo, supplementary information about chemical bonding and/or molecular distributions could be obtained by heavy-ion induced molecular desorption and a corresponding mass spectroscopy with Time-Of-Flight (TOF) mass spectrometer. As the method combines the use of heavy focused ions in MeV energy range and TOF Secondary Ion Mass Spectrometry, it is denoted as MeV SIMS.
At Jožef Stefan Institute, we constructed a linear TOF spectrometer and mount it to our multipurpose nuclear microprobe. A beam of 8MeV 35Cl7+ could be focused to a diameter of better than 3μm×3μm and pulsed by electrostatic deflection at the high-energy side of accelerator. TOF mass spectrometer incorporates an 1m long drift tube and a double stack microchannel plate (MCP) as a stop detector positioned at the end of the drift path. Secondary ions are focused at MCP using electrostatic cylindrical einzel lens.
Time of flight spectra are currently acquired with a single-hit time-to-digital converter. Pulsed ion beam produces a shower of secondary ions that are ejected from positively biased target and accelerated towards MCP. We start our time measurement simultaneously with the start of the beam pulse. Signal of the first ion hitting MCP is used to stop the time measurement. Standard pulses proportional to the time of flight are produced with time to analog converter (TAC) and fed into analog-to-digital converter to obtain a time histogram. To enable efficient detection of desorbed fragments with higher molecular masses, which are of particular interest, we recently implemented a state-of art Field Programmable Gate Array (FPGA)-based multi-hit TOF acquisition. To test the system we used focused 8MeV 35Cl7+ ion beam with pulse length of 180ns. Mass resolution of measured SIMS spectra, dominantly determined by the duration of the beam pulse, is in good agreement with resolution estimated from pulse length. With improved high-voltage switching ability that will enable beam pulses with duration of 50ns, a mass resolution of better than 500 is anticipated.
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