Ionome mapping and amino acid metabolome profiling of Phaseolus vulgaris L. seeds imbibed with computationally informed phytoengineered copper sulphide nanoparticles.

This study reports the effects of a computationally informed and avocado-seed mediated Phyto engineered CuS nanoparticles as fertilizing agent on the ionome and amino acid metabolome of Pinto bean seeds using both bench top and ion beam analytical techniques. Physico-chemical analysis of the Phyto engineered nanoparticles with scanning-electron microscopy, transmission electron microscopy, X-ray diffraction, and Fourier Transform Infrared Spectroscopy confirmed the presence of CuS nanoparticles. Molecular dynamics simulations to investigate the interaction of some active phytocompounds in avocado seeds that act as reducing agents with the nano-digenite further showed that 4-hydroxybenzoic acid had a higher affinity for interacting with the nanoparticle's surface than other active compounds. Seeds treated with the digenite nanoparticles exhibited a unique ionome distribution pattern as determined with external beam proton-induced X-ray emission, with hotspots of Cu and S appearing in the hilum and micropyle area that indicated a possible uptake mechanism via the seed coat. The nano-digenite also triggered a plant stress response by slightly altering seed amino acid metabolism. Ultimately, the nano-digenite may have important implications as a seed protective or nutritive agent as advised by its unique distribution pattern and effect on amino acid metabolism.

Three-dimensional Element-by-element Surface Topography Reconstruction of Compound Samples Using Multisegment Silicon Drift Detectors.

Elemental surface topography information in microscopic material characterization contributes to a better understanding of surfaces, interfaces, substrates, and their applications. Here, a general approach based on microbeam proton-induced X-ray emission (micro-PIXE) to reconstruct the three-dimensional (3D) elemental surface topography using the annular multisegment silicon drift detector has been demonstrated. The proposed method includes four main steps: acquiring four two-dimensional elemental concentration maps using the multichannel spectrometer, reconstructing the local inclination angle from the atomic model of ion-matter interaction, calculating the two independent topography gradient components, and numerical surface topography integration. In this study, the general algorithm to obtain the gradient components has been successfully tested on a four-segment configuration to reconstruct the 3D surface topography of compound alloys with different microstructure scales. In synchrotron and accelerator facilities dealing with elemental X-ray mapping where the development of customized multisegment detectors is needed, the introduced method is applicable to elemental surface/interface roughness reconstruction in microscale for cultural heritage samples, fusion plasma-facing materials, and microelectronic devices.

Tissue-specific calcium and magnesium allocation to explain differences in bulk concentration in leaves of one-year-old seedlings of two olive (Olea europaea L.) cultivars.

Olive tree (Olea europaea L.) leaves have recently been recognised as a valuable source in cosmetic and pharmaceutical industry as well as in preparation of health-supporting beverages. Little is known about the element composition of olive leaves and almost nothing about tissue-specific allocation of elements. Element composition and tissue-specific distribution were determined in leaves of two olive cultivars, Leccino and Istarska bjelica using micro-particle induced X-ray emission (micro-PIXE). In leaves of the Istarska bjelica cultivar larger bulk concentrations of potassium, sodium, molybdenum and boron, but smaller concentrations of calcium and magnesium were found than in leaves of the Leccino cultivar. Tissue-specific investigation revealed that larger concentration of calcium in epidermis and in leaf blade tissues (secondary veins, palisade and spongy mesophyll) contributed to the larger leaf bulk calcium concentration in the Leccino cultivar. For magnesium, all leaf tissues, except the bundle sheath cells and consequently the main vascular bundle, contributed to the larger bulk concentration in the Leccino cultivar. Potassium was not predominant in any of the leaf tissues examined, while sodium and molybdenum were below the limit of detection, and boron not detectable by micro-PIXE. The results indicate that sinks for calcium and magnesium are stronger in specific leaf tissues of the Leccino than of the Istarska bjelica cultivar. The new understanding of tissue-specific allocation of elements in leaves of olive will serve as a basis for detailed studies into the effects of foliar and/or soil fertilisers in olive.

Human Neocortex Layer Features Evaluated by PIXE, STIM, and STXM Techniques.

The human neocortex has a cytoarchitecture composed of six layers with an intrinsic organization that relates to afferent and efferent pathways for a high functional specialization. Various histological, neurochemical, and connectional techniques have been used to study these cortical layers. Here, we explore the additional possibilities of swift ion beam and synchrotron radiation techniques to distinguish cellular layers based on the elemental distributions and areal density pattern in the human neocortex. Temporal cortex samples were obtained from two neurologically normal adult men (postmortem interval: 6-12 h). A cortical area of 500 × 500 µm2 was scanned by a 3 MeV proton beam for elemental composition and areal density measurements using particle induced x-ray emission (PIXE) and scanning transmission ion microscopy (STIM), respectively. Zinc showed higher values in cortical layers II and V, which needs a critical discussion. Furthermore, the areal density decreased in regions with a higher density of pyramidal neurons in layers III and V. Scanning transmission X-ray microscopy (STXM) revealed the cellular density with higher lateral resolution than STIM, but not enough to distinguish each cortical lamination border. Our data describe the practical results of these approaches employing both X-ray and ion-beam based techniques for the human cerebral cortex and its heterogeneous layers. These results add to the potential approaches and knowledge of the human neocortical gray matter in normal tissue to develop improvements and address further studies on pathological conditions.

Total Knee Replacement with an Uncemented Porous Tantalum Tibia Component: A Failure Analysis.

Porous tantalum has been extensively used in orthopaedic surgery, including uncemented total knee arthroplasty (TKA). Favourable results were reported with earlier monobloc tibial components and the design evolved to modular implants. We aimed to analyse possible causes for extensive medial tibia bone loss, resulting in modular porous tantalum tibia baseplate fracture after primary TKA. Retrieved tissue samples were scanned with 3 MeV focused proton beam for Proton-Induced X-ray Emission (micro-PIXE) elemental analysis. Fractographic and microstructural analysis were performed by stereomicroscopy. A full 3D finite-element model was made for numerical analysis of stress-strain conditions of the tibial baseplate. Histological examination of tissue underneath the broken part of the tibial baseplate revealed dark-stained metal debris, which was confirmed by micro-PIXE to consist of tantalum and titanium. Fractographic analysis and tensile testing showed that the failure of the tibial baseplate fulfilled the criteria of a typical fatigue fracture. Microstructural analysis of the contact surface revealed signs of bone ingrowth in 22.5% of the surface only and was even less pronounced in the medial half of the tibial baseplate. Further studies are needed to confirm the responsibility of metal debris for an increased bone absorption leading to catastrophic tibial tray failure.

Molecular imaging of humain hair with MeV-SIMS: A case study of cocaine detection and distribution in the hair of a cocaine user.

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.

Revealing Inflammatory Indications Induced by Titanium Alloy Wear Debris in Periprosthetic Tissue by Label-Free Correlative High-Resolution Ion, Electron and Optical Microspectroscopy.

The metallic-associated adverse local tissue reactions (ALTR) and events accompanying worn-broken implant materials are still poorly understood on the subcellular and molecular level. Current immunohistochemical techniques lack spatial resolution and chemical sensitivity to investigate causal relations between material and biological response on submicron and even nanoscale. In our study, new insights of titanium alloy debris-tissue interaction were revealed by the implementation of label-free high-resolution correlative microscopy approaches. We have successfully characterized its chemical and biological impact on the periprosthetic tissue obtained at revision surgery of a fractured titanium-alloy modular neck of a patient with hip osteoarthritis. We applied a combination of photon, electron and ion beam micro-spectroscopy techniques, including hybrid optical fluorescence and reflectance micro-spectroscopy, scanning electron microscopy (SEM), Energy-dispersive X-ray Spectroscopy (EDS), helium ion microscopy (HIM) and micro-particle-induced X-ray emission (micro-PIXE). Micron-sized wear debris were found as the main cause of the tissue oxidative stress exhibited through lipopigments accumulation in the nearby lysosome. This may explain the indications of chronic inflammation from prior histologic examination. Furthermore, insights on extensive fretting and corrosion of the debris on nm scale and a quantitative measure of significant Al and V release into the tissue together with hydroxyapatite-like layer formation particularly bound to the regions with the highest Al content were revealed. The functional and structural information obtained at molecular and subcellular level contributes to a better understanding of the macroscopic inflammatory processes observed in the tissue level. The established label-free correlative microscopy approach can efficiently be adopted to study any other clinical cases related to ALTR.

Measuring the iron content of dopaminergic neurons in substantia nigra with MRI relaxometry.

In Parkinson's disease, the depletion of iron-rich dopaminergic neurons in nigrosome 1 of the substantia nigra precedes motor symptoms by two decades. Methods capable of monitoring this neuronal depletion, at an early disease stage, are needed for early diagnosis and treatment monitoring. Magnetic resonance imaging (MRI) is particularly suitable for this task due to its sensitivity to tissue microstructure and in particular, to iron. However, the exact mechanisms of MRI contrast in the substantia nigra are not well understood, hindering the development of powerful biomarkers. In the present report, we illuminate the contrast mechanisms in gradient and spin echo MR images in human nigrosome 1 by combining quantitative 3D iron histology and biophysical modeling with quantitative MRI on post mortem human brain tissue. We show that the dominant contribution to the effective transverse relaxation rate (R2*) in nigrosome 1 originates from iron accumulated in the neuromelanin of dopaminergic neurons. This contribution is appropriately described by a static dephasing approximation of the MRI signal. We demonstrate that the R2* contribution from dopaminergic neurons reflects the product of cell density and cellular iron concentration. These results demonstrate that the in vivo monitoring of neuronal density and iron in nigrosome 1 may be feasible with MRI and provide directions for the development of biomarkers for an early detection of dopaminergic neuron depletion in Parkinson's disease.

Temporal and Spatial Patterns of Zinc and Iron Accumulation during Barley (Hordeum vulgare L.) Grain Development.

Breeding and engineering of biofortified crops will benefit from a better understanding of bottlenecks controlling micronutrient loading within the seeds. However, few studies have addressed the changes in micronutrient concentrations, localization, and speciation occurring over time. Therefore, we studied spatial patterns of zinc and iron accumulation during grain development in two barley lines with contrasting grain zinc concentrations. Microparticle-induced-X-ray emission and laser ablation-inductively coupled plasma mass spectrometry were used to determine tissue-specific accumulation of zinc, iron, phosphorus, and sulfur. Differences in zinc accumulation between the lines were most evident in the endosperm and aleurone. A gradual decrease in zinc concentrations from the aleurone to the underlying endosperm was observed, while iron and phosphorus concentrations decreased sharply. Iron co-localized with phosphorus in the aleurone, whereas zinc co-localized with sulfur in the sub-aleurone. We hypothesize that differences in grain zinc are largely explained by the endosperm storage capacity. Engineering attempts should be targeted accordingly.

Secondary Ion Yield and Fragmentation of Biological Molecules by Employing 35Cl Primary Ions within the MeV Energy Domain.

MeV-SIMS is an emerging mass spectrometry imaging method that employs fast, heavy ions to desorb secondary molecules from the analyzed sample. High yields and low fragmentation rates of large molecules, associated with the dominating electronic sputtering process, make it particularly useful in biomedical research, where insight into the distribution of organic molecules is vital. Both yield and fragmentation of desorbed molecules in MeV-SIMS rely on characteristics of the primary ion but may also be impaired by poor instrumental settings. After utilizing secondary ion optics in the linear mass spectrometer at the micro-analytical center of the Jozef Stefan Institute, we demonstrate very efficient detection of secondary ions. As a result, the secondary ion yield, using such settings, solely depends on the species and the characteristics of the primary ion. In order to analyze the yield dependence on the primary ion energy, and the corresponding stopping power within the electronic excitation regime, we used a continuous electron multiplier detector to measure the primary ion current during each measurement of the mass spectra. Secondary ion yield as a function of the primary ion energy and charge is presented as well as fragmentation rates of organic molecules arginine and leu-enkephalin. Other influential instrumental drawbacks are also studied, and their effect on the results is discussed.

Effect of phosphorus supply on root traits of two Brassica oleracea L. genotypes.

BACKGROUND: Phosphorus (P) deficiency limits crop production worldwide. Crops differ in their ability to acquire and utilise the P available. The aim of this study was to determine root traits (root exudates, root system architecture (RSA), tissue-specific allocation of P, and gene expression in roots) that (a) play a role in P-use efficiency and (b) contribute to large shoot zinc (Zn) concentration in Brassica oleracea. RESULTS: Two B. oleracea accessions (var. sabellica C6, a kale, and var. italica F103, a broccoli) were grown in a hydroponic system or in a high-throughput-root phenotyping (HTRP) system where they received Low P (0.025 mM) or High P (0.25 mM) supply for 2 weeks. In hydroponics, root and shoot P and Zn concentrations were measured, root exudates were profiled using both Fourier-Transform-Infrared spectroscopy and gas-chromatography-mass spectrometry and previously published RNAseq data from roots was re-examined. In HTRP experiments, RSA (main and lateral root number and lateral root length) was assessed and the tissue-specific distribution of P was determined using micro-particle-induced-X-ray emission. The C6 accession had greater root and shoot biomass than the F103 accession, but the latter had a larger shoot P concentration than the C6 accession, regardless of the P supply in the hydroponic system. The F103 accession had a larger shoot Zn concentration than the C6 accession in the High P treatment. Although the F103 accession had a larger number of lateral roots, which were also longer than in the C6 accession, the C6 accession released a larger quantity and number of polar compounds than the F103 accession. A larger number of P-responsive genes were found in the Low P treatment in roots of the F103 accession than in roots of the C6 accession. Expression of genes linked with "phosphate starvation" was up-regulated, while those linked with iron homeostasis were down-regulated in the Low P treatment. CONCLUSIONS: The results illustrate large within-species variability in root acclimatory responses to P supply in the composition of root exudates, RSA and gene expression, but not in P distribution in root cross sections, enabling P sufficiency in the two B. oleracea accessions studied.

Arabidopsis halleri shows hyperbioindicator behaviour for Pb and leaf Pb accumulation spatially separated from Zn.

Lead (Pb) ranks among the most problematic environmental pollutants. Background contamination of soils is nearly ubiquitous, yet plant Pb accumulation is barely understood. In a survey covering 165 European populations of the metallophyte Arabidopsis halleri, several field samples had indicated Pb hyperaccumulation, offering a chance to dissect plant Pb accumulation. Accumulation of Pb was analysed in A. halleri individuals from contrasting habitats under controlled conditions to rule out aerial deposition as a source of apparent Pb accumulation. Several elemental imaging techniques were employed to study the spatial distribution and ligand environment of Pb. Regardless of genetic background, A. halleri individuals showed higher shoot Pb accumulation than A. thaliana. However, dose-response curves revealed indicator rather than hyperaccumulator behaviour. Xylem sap data and elemental imaging unequivocally demonstrated the in planta mobility of Pb. Highest Pb concentrations were found in epidermal and vascular tissues. Distribution of Pb was distinct from that of the hyperaccumulated metal zinc. Most Pb was bound by oxygen ligands in bidentate coordination. A. halleri accumulates Pb whenever soil conditions render Pb phytoavailable. Considerable Pb accumulation under such circumstances, even in leaves of A. thaliana, strongly suggests that Pb can enter food webs and may pose a food safety risk.

Contrasting allocation of magnesium, calcium and manganese in leaves of tea (Camellia sinensis (L.) Kuntze) plants may explain their different extraction efficiency into tea.

During tea preparation mineral elements are extracted from the dried leaves of tea (Camellia sinensis (L.) Kuntze) plants into the solution. Micro-particle induced X-ray emission was employed to investigate the spatial distribution of magnesium (Mg), calcium (Ca) and manganese (Mn) in the young and old leaves of tea plants grown in the absence and presence of aluminium (Al) in the substrate. Results revealed that in tea leaves the largest concentrations of Mg occurred in the epidermis, of Ca in oxalate crystals and of Mn in epidermis and oxalate crystals; there was a leaf-age effect on tissue-specific concentrations of Mg, Ca and Mn with all tissues of old leaves containing larger concentrations of Mg, Ca and Mn than young leaves; supplementation of substrate with Al reduced concentrations of Mg, Ca and Mn in the old leaves, and a link between the distribution of Mg, Ca and Mn in the tea leaves with the extraction efficiencies of these elements into the tea was possible. We conclude that old leaves of tea plants cultivated under conditions of low Al availability will have the largest concentrations of Mg, Ca and Mn and may represent most acceptable ingredient for the preparation of tea.

Nickel tolerance and toxicity mechanisms in the halophyte Sesuvium portulacastrum L. as revealed by Ni localization and ligand environment studies.

Halophytes are able to tolerate relatively high concentrations of hazardous metals in a growing substrate, what makes them suitable candidates for phytoremediation of metal-contaminated soils. In this work, we aimed to study the physiological responses of the halophyte Sesuvium portulacastrum L. to Ni, with main focus on Ni localization, compartmentation and ligand environment, to decipher Ni tolerance and toxicity mechanisms. Seedlings were grown in hydroponic nutrient solution containing 0, 25, 50 and 100 µM Ni as NiCl2 for 3 weeks. Ni localization in leaves was assessed by micro-proton-induced X-ray emission (micro-PIXE). Ni ligand environment was studied by Ni K-edge X-ray absorption near edge structure (XANES). In addition, Ni-soluble, weakly bound/exchangeable and insoluble leaf tissue fractions were determined by sequential extraction. Results show that S. portulacastrum is able to tolerate up to ~ 500 µg g-1 dry weight (DW) of Ni in the shoots without significant growth reduction. At higher Ni concentrations (> 50 µM Ni in nutrient solution), chloroses were observed due to the accumulation of Ni in photosynthetically active chlorenchyma as revealed by micro-PIXE. Water storage tissue represented the main pool for Ni storage. Incorporation of Ni into Ca-oxalate crystals was also observed in some specimens, conferring tolerance to high leaf Ni concentrations. The majority of Ni (> 70%) was found in soluble tissue fraction. Ni K XANES revealed Ni bound mainly to O- (55%) and N-ligands (45%). Ni toxicity at higher Ni levels was associated with Ni binding to amino groups of proteins in cytosol of chlorenchyma and increased level of lipid peroxidation. Proline levels also increased at high Ni exposures and were associated with Ni-induced oxidative stress and alteration of water regime.

Physico-elemental analysis of roasted organic coffee beans from Ethiopia, Colombia, Honduras, and Mexico using X-ray micro-computed tomography and external beam particle induced X-ray emission.

The physico-elemental profiles of commercially attained and roasted organic coffee beans from Ethiopia, Colombia, Honduras, and Mexico were compared using light microscopy, X-ray micro-computed tomography, and external beam particle induced X-ray emission. External beam PIXE analysis detected P, S, Cl, K, Ca, Ti, Mn, Fe, Cu, Zn, Br, Rb, and Sr in samples. Linear discriminant analysis showed that there was no strong association between elemental data and production region, whilst a heatmap combined with hierarchical clustering showed that soil-plant physico-chemical properties may influence regional elemental signatures. Physical trait data showed that Mexican coffee beans weighed significantly more than beans from other regions, whilst Honduras beans had the highest width. X-ray micro-computed tomography qualitative data showed heterogeneous microstructural features within and between beans representing different regions. In conclusion, such multi-dimensional analysis may present a promising tool in assessing the nutritional content and qualitative characteristics of food products such as coffee.

MeV-SIMS TOF Imaging of Organic Tissue with Continuous Primary Beam.

MeV-SIMS is an emerging mass spectrometry imaging method, which utilizes fast, heavy ions to desorb secondary molecules. High yields and low fragmentation rates of large molecules, associated with the electronic sputtering process, make it particularly useful in biomedical research, where insight into distribution of organic molecules is needed. Since the implementation of MeV-SIMS in to the micro-beam line at the tandem accelerator of Jozef Stefan Institute, MeV-SIMS provided some valuable observations on the distribution of biomolecules in plant tissue, as discussed by Jencic et al. (Nucl. Inst. Methods Phys. Res. B. 371, 205-210, 2016; Nucl. Inst. Methods Phys. Res. B. 404, 140-145, 2017). However, limited focusing ability of the chlorine ion beam only allowed imaging at the tissue level. In order to surpass shortcomings of the existing method, we introduced a new approach, where we employ a continuous, low-current primary beam. In this mode, we bombard thin samples with a steady chlorine ion flux of approx. 5000 ions/s. After desorbing molecules, chlorine ions penetrate through the thinly cut sample and trigger the time-of-flight "start" signal on a continuous electron multiplier detector, positioned behind the sample. Such bombardment is more effective than previously used pulsing-beam mode, which demanded several orders of magnitude higher primary ion beam currents. Sub-micrometer focusing of low-current primary ion beam allows imaging of biological tissue on a subcellular scale. Simultaneously, new time-of-flight acquisition approach also improves mass resolution by a factor of 5. Within the article, we compare the performance of both methods and demonstrate the application of continuous mode on biological tissue. We also describe the thin sample preparation protocol, necessary for measurements with low primary ion currents.

An ultra-stable gold-coordinated protein cage displaying reversible assembly.

Symmetrical protein cages have evolved to fulfil diverse roles in nature, including compartmentalization and cargo delivery1, and have inspired synthetic biologists to create novel protein assemblies via the precise manipulation of protein-protein interfaces. Despite the impressive array of protein cages produced in the laboratory, the design of inducible assemblies remains challenging2,3. Here we demonstrate an ultra-stable artificial protein cage, the assembly and disassembly of which can be controlled by metal coordination at the protein-protein interfaces. The addition of a gold (I)-triphenylphosphine compound to a cysteine-substituted, 11-mer protein ring triggers supramolecular self-assembly, which generates monodisperse cage structures with masses greater than 2 MDa. The geometry of these structures is based on the Archimedean snub cube and is, to our knowledge, unprecedented. Cryo-electron microscopy confirms that the assemblies are held together by 120 S-Aui-S staples between the protein oligomers, and exist in two chiral forms. The cage shows extreme chemical and thermal stability, yet it readily disassembles upon exposure to reducing agents. As well as gold, mercury(II) is also found to enable formation of the protein cage. This work establishes an approach for linking protein components into robust, higher-order structures, and expands the design space available for supramolecular assemblies to include previously unexplored geometries.

Physiological response and mineral elements accumulation pattern in Sesuvium portulacastrum L. subjected in vitro to nickel.

Sesuvium portulacastrum, a halophyte with high tolerance to heavy metals like Cd, Pb and Ni is considered for phytoremediation of metal contaminated saline soils. The tolerance to a selected metal ion could, by hypothesis, be stimulated through in vitro adaptation and regeneration of the plant. Seedlings obtained by in vitro micro-propagation, were exposed to 0, 25 and 50 µM Ni, as NiCl2, in agar-based medium for 30 days. Growth parameters, plant water content, the concentration of photosynthetic pigments, proline and malondialdehyde (MDA) concentrations were determined. Nickel and nutrients distribution in leaves was studied by micro-Proton-Induced-X-ray-Emission (µ-PIXE). The results showed that Ni was mainly accumulated in vascular bundles, next in water storage tissues and chlorenchyma. Ni concentrations in chlorenchyma increased with increasing Ni in culturing medium, in direct relation to decrease of photosynthetic pigments and increase of oxidative stress. As compared to control plants, Ni induced substantial increase in MDA and proline accumulation. Plants exposed to 50 µM Ni accumulated up to 650 µg g-1 of Ni in the shoots, exhibiting chlorosis and necrosis and a drastically reduced plant growth. Perturbations in uptake and distribution of nutrients were observed, inducing mineral deficiency, probably through membrane leakage. The mineral nutrient disturbances induced by Ni could be highly implicated in the restriction of S. portulacastrum development under the acute 50 µM Ni level.

Nano Zero-Valent Iron Mediated Metal(loid) Uptake and Translocation by Arbuscular Mycorrhizal Symbioses.

Nano zero-valent iron (nZVI) has great potential in the remediation of metal(loid)-contaminated soils, but its efficiency in metal(loid) stabilization in the plant-microbe continuum is unclear. This study investigated nZVI-mediated metal(loid) behavior in the arbuscular mycorrhizal (AM) fungal-maize ( Zea mays L.) plant association. Plants with AM fungal inoculation were grown in metal(loid)- (mainly Zn and Pb) contaminated soils (Litavka River, Czech Republic) amended with/without 0.5% (w/w) nZVI. The results showed that nZVI decreased plant metal(loid) uptake but inhibited AM development and its function in metal(loid) stabilization in the rhizosphere. AM fungal inoculation alleviated the physiological stresses caused by nZVI and restrained nZVI efficiency in reducing plant metal(loid) uptake. Micro proton-induced X-ray emission (µ-PIXE) analysis revealed the sequestration of Zn (possibly through binding to thiols) by fungal structures in the roots and the precipitation of Pb and Cu in the mycorrhizal root rhizodermis (possibly by Fe compounds originated from nZVI). XRD analyses further indicated that Pb/Fe mineral transformations in the rhizosphere were influenced by AM and nZVI treatments. The study revealed the counteractive effects of AM and nZVI on plant metal(loid) uptake and uncovered details of metal(loid) behavior in the AM fungal-root-nZVI system, calling into question about nZVI implementation in mycorrhizospheric systems.

Why do some titanium-alloy total hip arthroplasty modular necks fail?

BACKGROUND: Increased modularity in total hip arthroplasty (THA) through extra junction between the neck and the femoral stem is gaining popularity among orthopaedic community. However, the advantage of the additional junction is shadowed by an increased risk of mechanical failure. The aim of this study was to describe the exact mechanism of fracture of the modular femoral neck in an uncemented stem. METHODS: Clinical, metallurgical, and mechanical analysis including finite-element modelling and elemental-sensitive tissue analysis with the micro-PIXE method was performed on two patients treated with fully modular primary THA made from Titanium alloy of the same oval taper-cone design. In patient A revision was performed 7.8 years after the unilateral primary procedure because of modular femoral neck fracture, while patient B was left-side revised 15 years after the bilateral primary procedure because of aseptic loosening of the femoral stem. RESULTS: Body weight was 30% higher and the arm of implanted modular femoral neck was 51% longer in patient A compared to patient B. Therefore, the stress ratio on the modular femoral neck of patient A was calculated to be 2.45 times higher than in patient B, preventing cold welding and producing taper damage and degradation at the neck-stem junction. Large clusters of metallic debris containing Titanium and Vanadium from the alloy were present in the periprosthetic soft tissues of patient A. CONCLUSIONS: Patients with higher body mass index treated with fully modular Ti-alloy THA may be at increased risk to experience catastrophic failure of the device. Orthopaedic surgeons should avoid using long necks whenever possible, as these are especially prone to develop a vicious circle starting with the fretting process and crevice corrosion at the taper-cone connection, leading to crack initiation and crack propagation, accelerated by the increased vulnerability of the Ti-alloy in biologic media, ultimately ending as fracture at the typical site. Serum Ti concentration may represent a rough estimation of taper degradation and patients with elevated levels should be warned and followed accordingly.