Nickel oxide nanoparticles induce cell wall modifications, root anatomical changes, and nitrosative signaling in ecotypes of Ni hyperaccumulator Odontarrhena lesbiaca.

The industrial application and environmental release of nickel oxide NPs (NiO NPs) is increasing, but the details of their relationship with plants are largely unknown. In this work, the cellular, tissue, organ, and molecular level responses of three ecotypes of Ni hyperaccumulator Odontarrhena lesbiaca grown in the presence of high doses of NiO NP (250 mg/L and 500 mg/L) were studied. All three ecotypes showed a similar accumulation of Ni in the presence of nano Ni, and in the case of NiO NPs, the root-to-shoot Ni translocation was slighter compared to the bulk Ni. In all three ecotypes, the walls of the root cells effectively prevented internalization of NiO NPs, providing cellular defense against Ni overload. Exposure to NiO NP led to an increase in cortex thickness and the deposition of lignin-suberin and pectin in roots, serving as a tissue-level defense mechanism against excessive Ni. Exposure to NiO NP did not modify or cause a reduction in some biomass parameters of the Ampeliko and Loutra ecotypes, while it increased all parameters in Olympos. The free salt form of Ni exerted more negative effects on biomass production than the nanoform, and the observed effects of NiO NPs can be attributed to the release of Ni ions. Nitric oxide and peroxynitrite levels were modified by NiO NPs in an ecotype-dependent manner. The changes in the abundance and activity of S-nitrosoglutathione reductase protein triggered by NiO NPs suggest that the enzyme is regulated by NiO NPs at the post-translational level. The NiO NPs slightly intensified protein tyrosine nitration, and the slight differences between the ecotypes were correlated with their biomass production in the presence of NiO NPs. Overall, the Odontarrhena lesbiaca ecotypes exhibited tolerance to NiO NPs at the cellular, tissue, organ/organism and molecular levels, demonstrating various defense mechanisms and changes in the metabolism of reactive nitrogen species metabolism and nitrosative protein modification.

Design, Optimization, and Application of a 3D-Printed Polymer Sample Introduction System for the ICP-MS Analysis of Nanoparticles and Cells.

Commonly used sample introduction systems for inductively coupled plasma mass spectrometry (ICP-MS) are generally not well-suited for single particle ICP-MS (spICP-MS) applications due to their high sample requirements and low efficiency. In this study, the first completely 3D-printed, polymer SIS was developed to facilitate spICP-MS analysis. The system is based on a microconcentric pneumatic nebulizer and a single-pass spray chamber with an additional sheath gas flow to further facilitate the transport of larger droplets or particles. The geometry of the system was optimized using numerical simulations. Its aerosol characteristics and operational conditions were studied via optical particle counting and a course of spICP-MS measurements, involving nanodispersions and cell suspensions. In a comparison of the performance of the new and the standard (quartz microconcentric nebulizer plus a double-pass spray chamber) systems, it was found that the new sample introduction system has four times higher particle detection efficiency, significantly better signal-to-noise ratio, provides ca. 20% lower size detection limit, and allows an extension of the upper limit of transportable particle diameters to about 25 µm.

A Calibration-Free Optical Emission Spectroscopic Method to Determine the Composition of a Spark Discharge Plasma Used for AuAg Binary Nanoparticle Synthesis.

Spark discharge generators (SDGs) employ controlled gaseous environments to induce spark ablation of non-insulating electrodes, resulting in the formation of various nanostructures in the gas phase. The method offers technological advantages such as continuous particle production, scalable yield, and minimal waste. Additionally, the versatility of the process enables the generation of alloy nanoparticles from various material combinations, including immiscible ones. In order to fully exploit its potential, understanding the atomic mixing process during electrode ablation, particularly in the case of dissimilar electrodes, is crucial. Temporally and spatially resolved optical emission spectroscopy (OES) has been previously demonstrated as an effective characterization tool for spark plasmas in SDGs. However, to gain a deeper insight into the vapor mixing process, it is essential to quantitatively determine the plasma composition in both space and time. This paper introduces a calibration-free OES-based method tailored for spark plasmas utilized in binary nanoparticle generation. The method introduces the so-called multi-element combinatory Boltzmann plots, which use intensity ratios of emission atomic lines from different materials, allowing for the direct estimation of total number concentration ratios. The approach is tested using synthetic spectra and validated with experimental spark spectra obtained near an alloyed gold-silver (AuAg) electrode with a known composition. The study demonstrates the capabilities and robustness of the proposed method, with a focus on the AuAg system due to its significance in plasmonic research and frequent synthesis using spark ablation.

Selenate triggers diverse oxidative responses in Astragalus species with diverse selenium tolerance and hyperaccumulation capacity.

Selenium (Se) hyperaccumulators are capable of uptake and tolerate high Se dosages. Excess Se-induced oxidative responses were compared in Astragalus bisulcatus and Astragalus cicer. Plants were grown on media supplemented with 0, 25 or 75 µM selenate for 14 days. Both A. bisulcatus and A. cicer accumulated >2000 µg/g dry weight Se to the shoot but the translocation factors of A. cicer were below 1 suggesting its non hyperaccumulator nature. A. cicer showed Se sensitivity indicated by reduced seedling fresh weight, root growth and root apical meristem viability, altered element homeostasis in the presence of Se. In Se-exposed A. bisulcatus, less toxic organic Se forms (mainly MetSeCys, γ-Glu-MetSeCys, and a selenosugar) dominated, while these were absent from A. cicer suggesting that the majority of the accumulated Se may be present as inorganic forms. The glutathione-dependent processes were more affected, while ascorbate levels were not notably influenced by Se in either species. Exogenous Se triggered more intense accumulation of malondialdehyde in the sensitive A. cicer compared with the tolerant A. bisulcatus. The extent of protein carbonylation in the roots of the 75 µM Se-exposed A. cicer exceeded that of A. bisulcatus indicating a correlation between selenate sensitivity and the degree of protein carbonylation. Overall, our results reveal connection between oxidative processes and Se sensitivity/tolerance/hyperaccumulation and contribute to the understanding of the molecular responses to excess Se.

Quantitative elemental mapping of biological tissues by laser-induced breakdown spectroscopy using matrix recognition.

The present study demonstrates the importance of converting signal intensity maps of organic tissues collected by laser-induced breakdown spectroscopy (LIBS) to elemental concentration maps and also proposes a methodology based on machine learning for its execution. The proposed methodology employs matrix-matched external calibration supported by a pixel-by-pixel automatic matrix (tissue type) recognition performed by linear discriminant analysis of the spatially resolved LIBS hyperspectral data set. On a swine (porcine) brain sample, we successfully performed this matrix recognition with an accuracy of 98% for the grey and white matter and we converted a LIBS intensity map of a tissue sample to a correct concentration map for the elements Na, K and Mg. Found concentrations in the grey and white matter agreed the element concentrations published in the literature and our reference measurements. Our results revealed that the actual concentration distribution in tissues can be quite different from what is suggested by the LIBS signal intensity map, therefore this conversion is always suggested to be performed if an accurate concentration distribution is to be assessed.

Continuous spark plasma synthesis of Au/Co binary nanoparticles with tunable properties.

We present here a scalable and environmentally friendly gas phase technique employing atmospheric pressure electrical spark discharge plasmas for the production of Au/Co binaries, an effective catalyst system for the decomposition of hydrogen-rich compounds, such as ammonium borane. We demonstrate that Au/Co alloy nanoparticles can be produced via the spark plasma-based technique. The possibility of varying the morphology and phase structure via real time heat treatment of the generated aerosol to form Au/Co/CoO particles with continuous control over a wide particle compositional range (from 24 to 64 at.% [Co]/([Co] + [Au]) content) is also demonstrated. Since our spark-based approach is proven to be capable of providing reasonable particle yields, these results may contribute to the transition of lab-scale, nanocatalyst-based hydrogen storage systems to real world applications.

The Effect of Femtosecond Laser Irradiation and Plasmon Field on the Degree of Conversion of a UDMA-TEGDMA Copolymer Nanocomposite Doped with Gold Nanorods.

In this work, the effects of femtosecond laser irradiation and doping with plasmonic gold nanorods on the degree of conversion (DC) of a urethane dimethacrylate (UDMA)-triethylene glycol dimethacrylate (TEGDMA) nanocomposite were investigated. The UDMA-TEGDMA photopolymer was prepared in a 3:1 weight ratio and doped with dodecanethiol- (DDT) capped gold nanorods of 25 × 75 or 25 × 85 nm nominal diameter and length. It was found that the presence of the gold nanorods alone (without direct plasmonic excitation) can increase the DC of the photopolymer by 6-15%. This increase was found to be similar to what could be achieved with a control heat treatment of 30 min at 180 °C. It was also shown that femtosecond laser impulses (795 nm, 5 mJ pulse energy, 50 fs pulse length, 2.83 Jcm-2 fluence), applied after the photopolymerization under a standard dental curing lamp, can cause a 2-7% increase in the DC of undoped samples, even after thermal pre-treatment. The best DC values (12-15% increase) were obtained with combined nanorod doping and subsequent laser irradiation close to the plasmon resonance peak of the nanorods (760-800 nm), which proves that the excited plasmon field can directly facilitate double bond breakage (without thermoplasmonic effects due to the short pulse length) and increase the crosslink density independently from the initial photopolymerization process.

Towards Automated Classification of Zooplankton Using Combination of Laser Spectral Techniques and Advanced Chemometrics.

Zooplankton identification has been the subject of many studies. They are mainly based on the analysis of photographs (computer vision). However, spectroscopic techniques can be a good alternative due to the valuable additional information that they provide. We tested the performance of several chemometric techniques (principal component analysis (PCA), non-negative matrix factorisation (NMF), and common dimensions and specific weights analysis (CCSWA of ComDim)) for the unsupervised classification of zooplankton species based on their spectra. The spectra were obtained using laser-induced breakdown spectroscopy (LIBS) and Raman spectroscopy. It was convenient to assess the discriminative power in terms of silhouette metrics (Sil). The LIBS data were substantially more useful for the task than the Raman spectra, although the best results were achieved for the combined LIBS + Raman dataset (best Sil = 0.67). Although NMF (Sil = 0.63) and ComDim (Sil = 0.39) gave interesting information in the loadings, PCA was generally enough for the discrimination based on the score graphs. The distinguishing between Calanoida and Euphausiacea crustaceans and Limacina helicina sea snails has proved possible, probably because of their different mineral compositions. Conversely, arrow worms (Parasagitta elegans) usually fell into the same class with Calanoida despite the differences in their Raman spectra.

Distinct Uptake Routes Participate in Silver Nanoparticle Engulfment by Earthworm and Human Immune Cells.

The consequences of engineered silver nanoparticle (AgNP) exposure and cellular interaction with the immune system are poorly understood. The immunocytes of the Eisenia andrei earthworm are frequently applied in ecotoxicological studies and possess functional similarity to vertebrate macrophages. Hence, we characterized and compared the endocytosis mechanisms for the uptake of 75 nm AgNPs by earthworm coelomocytes, human THP-1 monocytes, and differentiated THP-1 (macrophage-like) cells. Our results indicate that microtubule-dependent, scavenger-receptor, and PI3K signaling-mediated macropinocytosis are utilized during AgNP engulfment by human THP-1 and differentiated THP-1 cells. However, earthworm coelomocytes employ actin-dependent phagocytosis during AgNPs uptake. In both human and earthworm immunocytes, AgNPs were located in the cytoplasm, within the endo-/lysosomes. We detected that the internalization of AgNPs is TLR/MyD88-dependent, also involving the bactericidal/permeability-increasing protein (BPI) in the case of human immunocytes. The exposure led to decreased mitochondrial respiration in human immunocytes; however, in coelomocytes, it enhanced respiratory parameters. Our findings provide more data about NP trafficking as nano-carriers in the nanomedicine field, as well as contribute to an understanding of the ecotoxicological consequences of nanoparticle exposure.

Multifunctional microfluidic chips for the single particle inductively coupled plasma mass spectrometry analysis of inorganic nanoparticles.

This study aimed at exploiting the so far unexploited potential of carrying out on-line sample pretreatment steps on microfluidic chips for single particle inductively coupled plasma mass spectrometry (spICP-MS) measurements, and demonstrating their ability to practically facilitate most of the simpler tasks involved in the spICP-MS analysis of nanoparticles. For this purpose, polydimethylsiloxane microfluidic chips, capable of high-range dilution and sample injection were made by casting, using high-precision, 3D-printed molds. Optimization of their geometry and functions was done by running several hydrodynamic simulations and by gravimetric, fluorescence enhanced microscope imaging and solution-based ICP-MS experiments. On the optimized microfluidic chips, several experiments were done, demonstrating the benefits of the approach and these devices, such as the determination of nanoparticle concentration using only a few tens of microliters of sample, elimination of solute interferences by dilution, solution-based size calibration and characterisation of binary nanoparticles. Due to the unique design of the chips, they can be linked together to extend the dilution range of the system by more than a magnitude per chip. This feature was also demonstrated in applications requiring multiple-magnitude dilution rates, when two chips were sequentially coupled.

A novel approach for discovering correlations between elemental and molecular composition using laser-based spectroscopic techniques.

Modern analytical techniques, including laser-induced breakdown spectroscopy (LIBS) and Raman spectroscopy, yield multidimensional data, which are most efficiently used in conjunction with chemometric techniques, including multi-block algorithms. In this study, we use several algorithms for the processing of laser-induced breakdown and Raman spectra of zooplankton organisms, which are found to accumulate lithium for an unknown reason. Correlations between elemental and molecular composition of zooplankton have been found. We studied 29 samples: crustaceans, arrow worms, and sea snails. The obtained spectra were examined by principal component analysis (PCA), non-negative matrix factorization (NMF), consensus PCA (CPCA), and analysis of common components and specific weights (CCSWA, or ComDim). LIBS spectra are more sensitive towards taxonometric differences than Raman spectra. All the algorithms gave similar results, although still differing in details. Data fusion revealed a number of relationships, including the correlation of Li with potassium (R = 0.83, n = 14), with Raman bands of carotenoids (R = 0.89, n = 11) and tryptophan (R = 0.94, n = 9). The correlations were most pronounced in light-coloured parts of the inhomogeneous biological material. Ratios of fatty acids are associated with Li concentration if above 200 mg kg-1. Valine is also related to the Li accumulation. Thus, it is shown that the combination of LIBS and Raman spectroscopy, followed by appropriate mathematical treatment, is a convenient tool for comprehensive studies of environmental objects.

Qualitative Analysis of Glass Microfragments Using the Combination of Laser-Induced Breakdown Spectroscopy and Refractive Index Data.

We have successfully demonstrated that although there are significant analytical challenges involved in the qualitative discrimination analysis of sub-mm sized (microfragment) glass samples, the task can be solved with very good accuracy and reliability with the multivariate chemometric evaluation of laser-induced breakdown spectroscopy (LIBS) data or in combination with pre-screening based on refractive index (RI) data. In total, 127 glass samples of four types (fused silica, flint, borosilicate and soda-lime) were involved in the tests. Four multivariate chemometric data evaluation methods (linear discrimination analysis, quadratic discrimination analysis, classification tree and random forest) for LIBS data were evaluated with and without data compression (principal component analysis). Classification tree and random forest methods were found to give the most consistent and most accurate results, with classifications/identifications correct in 92 to 99% of the cases for soda-lime glasses. The developed methods can be used in forensic analysis.

One-step fabrication of fiber optic SERS sensors via spark ablation.

Spark ablation, a versatile, gas-phase physical nanoparticle synthesis method was employed to fabricate fiber-optic surface enhanced Raman scattering (SERS) sensors in a simple single-step process. We demonstrate that spark-generated silver nanoparticles can be simply deposited onto a fiber tip by means of a modified low-pressure inertial impactor, thus providing significant surface enhancement for fiber-based Raman measurements. The surface morphology of the produced sensors was characterized along with the estimation of the enhancement factor and the inter- and intra-experimental variation of the measured Raman spectrum as well as the investigation of the concentration dependence of the SERS signal. The electric field enhancement over the deposited silver nanostructure was simulated in order to facilitate the better understanding of the performance of the fabricated SERS sensors. A potential application in the continuous monitoring of a target molecule was demonstrated on a simple model system.

Full range tuning of the composition of Au/Ag binary nanoparticles by spark discharge generation.

Gold/silver bimetallic nanoparticles still attract extensive interest due to their favorable properties e.g. in plasmonics or catalysis. We present here a facile and robust way for the production of clean Au/Ag binary nanoparticles (BNPs) with a total control over the composition via the spark discharge nanoparticle generation technique. With the application of pure Ag and Au electrodes, a tuning range of 55 to 90% Au content was achieved, but this can be further extended to the full 0 to 100% range by using a couple of alloyed electrodes. An added benefit of the approach is that either the concentration or the mean particle size can be kept constant at every composition by adjusting the generator parameters. Based on the systematic experimental data collected, a semi-empirical model for the prediction of the Au/Ag BNP composition was also developed. This model was used to calculate the theoretically achievable Au/Ag composition at a given spark parameter set in the parameter range most commonly used in the literature.

Modulation of the catalytic activity of a metallonuclease by tagging with oligohistidine.

Peptide tags are extensively used for affinity purification of proteins. In an optimal case, these tags can be completely removed from the purified protein by a specific protease mediated hydrolysis. However, the interactions of these tags with the target protein may also be utilized for the modulation of the protein function. Here we show that the C-terminal hexahistidine (6 × His) tag can influence the catalytic activity of the nuclease domain of the Colicin E7 metallonuclease (NColE7) used by E. coli to kill competing bacteria under stress conditions. This enzyme non-specifically cleaves the DNA that results in cytotoxicity. We have successfully cloned the genes of NColE7 protein and its R447G mutant into a modified pET-21a DNA vector fusing the affinity tag to the protein upon expression, which would be otherwise not possible in the absence of the gene of the Im7 inhibitory protein. This reflects the inhibitory effect of the 6 × His fusion tag on the nuclease activity, which proved to be a complex process via both coordinative and non-specific steric interactions. The modulatory effect of Zn2+ ion was observed in the catalytic activity experiments. The DNA cleavage ability of the 6 × His tagged enzyme was first enhanced by an increase of metal ion concentration, while high excess of Zn2+ ions caused a lower rate of the DNA cleavage. Modelling of the coordinative effect of the fusion tag by external chelators suggested ternary complex formation instead of removal of the metal ion from the active center.

Deuterium analysis by inductively coupled plasma mass spectrometry using polyatomic species: An experimental study supported by plasma chemistry modeling.

A new analytical method is proposed for the determination of deuterium (D) by ICP-MS. The method is based on the use of the signal from hydrogen-containing polyatomic ions formed in the inductively coupled plasma. Prior to analytical experiments, a theoretical study was performed to assess the concentration of polyatomic species present in an equilibrium Ar-O-D-H plasma, as a function of temperature and stoichiometric composition. It was established that the highest sensitivity and linearity measurement of D concentration in a wide range can be achieved by monitoring the ions of D2 and ArD, at masses 4 and 42, respectively. Results of the calculations are in good agreement with the experiments. Signal stability, spectral interferences, as well as the effect of plasma parameters were also assessed. Under optimized conditions, the limit of detection (LOD) was found to be 3 ppm atom fraction for deuterium when measured as ArD (in calcium and potassium free water), or 78 ppm when measured as D2. The achieved LOD values and the 4 to 5 orders of magnitude dynamic range easily allow the measurement of deuterium concentrations at around or above the natural level, up to nearly 100% (or 1 Mio ppm) in a standard quadrupole ICP-MS instrument. An even better performance is expected from the method in high resolution ICP-MS instruments equipped with low dead volume sample introduction systems.

Analysis and Classification of Liquid Samples Using Spatial Heterodyne Raman Spectroscopy.

Spatial heterodyne spectroscopy (SHS) is used for quantitative analysis and classification of liquid samples. SHS is a version of a Michelson interferometer with no moving parts and with diffraction gratings in place of mirrors. The instrument converts frequency-resolved information into a spatially resolved one and records it in the form of interferograms. The back-extraction of spectral information is done by the fast Fourier transform. A SHS instrument is constructed with the resolving power 5000 and spectral range 522-593 nm. Two original technical solutions are used as compared to previous SHS instruments: the use of a high-frequency diode-pumped solid-state laser for excitation of Raman spectra and a microscope-based collection system. Raman spectra are excited at 532 nm at the repetition rate 80 kHz. Raman shifts between 330 cm-1 and 1600 cm-1 are measured. A new application of SHS is demonstrated: for the first time, it is used for quantitative Raman analysis to determine concentrations of cyclohexane in isopropanol and glycerol in water. Two calibration strategies are employed: univariate based on the construction of a calibration plot and multivariate based on partial least squares regression. The detection limits for both cyclohexane in isopropanol and glycerol in water are at a 0.5 mass% level. In addition to the Raman-SHS chemical analysis, classification of industrial oils (biodiesel, poly(1-decene), gasoline, heavy oil IFO380, polybutenes, and lubricant) is performed using the Raman-fluorescence spectra of the oils and principal component analysis. The oils are easily discriminated showing distinct non-overlapping patterns in the principal component space.

Hg2+ and Cd2+ binding of a bioinspired hexapeptide with two cysteine units constructed as a minimalistic metal ion sensing fluorescent probe.

Hg2+ and Cd2+ complexation of a short hexapeptide, Ac-DCSSCY-NH2 (DY), was studied by pH-potentiometry, UV and NMR spectroscopy and fluorimetry in aqueous solutions and the Hg2+-binding ability of the ligand was also described in an immobilized form, where the peptides were anchored to a hydrophilic resin. Hg2+ was demonstrated to form a 1 : 1 complex with the ligand even at pH = 2.0 while Cd2+ coordination by the peptide takes place only above pH ∼ 3.5. Both metal ions form bis-ligand complexes by the coordination of four Cys-thiolates at ligand excess above pH ∼ 5.5 (Cd2+) and 7.0 (Hg2+). Fluorescence studies demonstrated a Hg2+ induced concentration-dependent quenching of the Tyr fluorescence until a 1 : 1 Hg2+ : DY ratio. The fluorescence emission intensity decreases linearly with the increasing Hg2+ concentration in a range of over two orders of magnitude. The fact that this occurs even in the presence of 1.0 eq. of Cd2+ per ligand reflects a complete displacement of the latter metal ion by Hg2+ from its peptide-bound form. The immobilized peptide was also shown to bind Hg2+ very efficiently even from samples at pH = 2.0. However, the existence of lower affinity binding sites was also demonstrated by binding of more than 1.0 eq. of Hg2+ per immobilized DY molecule under Hg2+-excess conditions. Experiments performed with a mixture of four metal ions, Hg2+, Cd2+, Zn2+ and Ni2+, indicate that this molecular probe may potentially be used in Hg2+-sensing systems under acidic conditions for the measurement of µM range concentrations.

Synthesis and spectroscopic characterization of novel GFP chromophore analogues based on aminoimidazolone derivatives.

In order to improve the fluorescence properties of the green fluorescent protein chromophore, p­HOBDI ((5­(4­hydroxybenzylidene)­2,3­dimethyl­3,5­dihydro­4H­imidazol­4­one), sixteen dihydroimidazolone derivates were synthesized from thiohydantoin and arylaldehydes. The synthesis developed is an efficient, novel, one-pot procedure. The study provides a detailed description of the spectroscopic characteristics of the newly synthesized compounds, using p­HOBDI as a reference. The new compounds all exhibited significantly stronger fluorescence than p­HOBDI, up to 28 times higher quantum yields. An experimental and theoretical investigation of the relationship of the fluorescence properties with the molecular structure was also carried out. A good correlation was found between the emission wavenumber and the Hammett constant of the functional group, which suggests the intermolecular charge transfer (ICT) mechanism between the aromatic groups.

Designed Pt Promoted 3D Mesoporous Co3O4 Catalyst in CO2 Hydrogenation.

Controlled size Pt nanoparticles were anchored onto the surface of 3D mesoporous cobalt-oxide support and was tested in CO2 hydrogenation reactions compared to commercial cobalt-oxide supported Pt nanoparticles prepared by the wet impregnation method as well as SBA-15 silica supported nanoparticles. Designed Pt/mesoporous cobalt-oxide catalysts showed the highest activity as well as the highest methane selectivity. Such catalyst was active at 573 K, while other catalysts showed activity >673 K.