Course of Plasmodium infection studied using 2D-COS on human erythrocytes.

BACKGROUND: The threat of malaria is still present in the world. Recognizing the type of parasite is important in determining a treatment plan. The golden routine involves microscopic diagnostics of Giemsa-stained thin blood smears, however, alternative methods are also constantly being sought, in order to gain an additional insight into the course of the disease. Spectroscopic methods, e.g., Raman spectroscopy, are becoming increasingly popular, due to the non-destructive nature of these techniques. METHODS: The study included patients hospitalized for malaria caused by Plasmodium falciparum or Plasmodium vivax, in the Department of Infectious Diseases at the University Hospital in Krakow, Poland, as well as healthy volunteers. The aim of this study was to assess the possibility of using Raman spectroscopy and 2D correlation (2D-COS) spectroscopy in understanding the structural changes in erythrocytes depending on the type of attacking parasite. EPR spectroscopy and two-trace two-dimensional (2T2D) correlation was also used to examine the specificity of paramagnetic centres found in the infected human blood. RESULTS: Two-dimensional (2D) correlation spectroscopy facilitates the identification of the hidden relationship, allowing for the discrimination of Raman spectra obtained during the course of disease in human red blood cells, infected by P. falciparum or P. vivax. Synchronous cross-peaks indicate the processes taking place inside the erythrocyte during the export of the parasite protein towards the cell membrane. In contrast, moieties that generate asynchronous 2D cross-peaks are characteristic of the respective ligand-receptor domains. These changes observed during the course of the infection, have different dynamics for P. falciparum and P. vivax, as indicated by the asynchronous correlation cross-peaks. Two-trace two-dimensional (2T2D) spectroscopy, applied to EPR spectra of blood at the beginning of the infection, showed differences between P. falciparum and P. vivax. CONCLUSIONS: A unique feature of 2D-COS is the ability to discriminate the collected Raman and EPR spectra. The changes observed during the course of a malaria infection have different dynamics for P. falciparum and P. vivax, indicated by the reverse sequence of events. For each type of parasite, a specific recycling process for iron was observed in the infected blood.

The effect of UV-B irradiation on structural and functional properties of corn and potato starches and their components.

In this work, UV-B irradiation, an environmentally friendly modification method, was applied to corn and potato carbohydrates. The influence of irradiation on starch properties was compared with that observed for starch oxidation with NaClO. The changes in the structures of starch carbohydrates were investigated by X-ray diffraction (XRD), electron paramagnetic resonance (EPR) and chromatographic methods. Functional properties such as viscosity, water binding capacity and solubility as well as pasting characteristics and thermodynamic parameters of gelatinization, resulting from structural changes were determined. UV-B irradiation was found to be a milder modification method than chemical oxidation. The potato carbohydrates, especially amylose, appeared to be more susceptible to changes upon irradiation, whereas corn ones, particularly amylopectin, were more stable and their properties changed to a smaller extent. Similarly, functional properties were not significantly influenced by UV-B treatment. EPR studies revealed the mechanism of depolymerization of starch carbohydrates via formation of stable carbon-centered radicals.

The effect of thermal and irradiation treatments on structural and physicochemical properties of octenyl succinate maize starches.

The aim of this paper was to evaluate the effect of thermal treatment and UV irradiation on structural and physicochemical properties of octenyl succinate (OS) maize starches differing in content of OS groups (0.76%-2.38%). X-ray diffraction (XRD), gel permeation chromatography (GPC), and electron paramagnetic resonance (EPR) spectroscopy were used to estimate structural characteristics of octenyl succinate starches and their physically treated counterparts. Starch functional properties were also determined. XRD spectra and hydrodynamic volume distribution revealed that growth of octenyl succinate groups' content resulted in stabilization of starch structure, which was confirmed by processes of radical formation, stimulated by thermal and UV treatments. It was found, on the basis of measurements of water binding capacity, water solubility and intrinsic viscosity, that OSA addition changed physicochemical properties of starch and influenced the course of physical modifications. It was established that temperature affected starch properties to a greater extent than UV.

High-Molecular-Weight Polyampholytes Synthesized via Daylight-Induced, Initiator-Free Radical Polymerization of Renewable Itaconic Acid.

Herein, it is reported for the first time that when mixed with choline chloride, itaconic acid (IA), normally a low-reactive vinyl monomer, undergoes initiator-free radical polymerization under normal daylight. Furthermore, the process results in the formation of abnormally high-molecular-weight poly(itaconic acid) derivatives with Mw greater than ≈800 000 g mol-1 . Detailed 1D/2D NMR studies indicate that the polymers have two types of ionizable moieties, that is, anionic carboxylic and cationic choline ester groups in an average molar ratio of 12:1. Potentiometric titration shows polyampholyte behavior of the polymers. Tentative mechanistic studies reveal that the daylight-induced polymerization is initiated by species generated via interactions of near UV light with IA. However, EPR findings show that choline also participates in secondary radical reactions. The obtained polyampholytes are useful bio-based materials for fast and straightforward fabrication of polymer-clay nanocomposite hydrogels with excellent mechanical properties.

Translocation of elements and sugars in wheat genotypes at vegetative and generative stages under continuous selenium exposure.

BACKGROUND: Biofortification with selenium (Se) elevates its concentration in feed and fodder plants and helps to prevent health problems in animals and humans. The aim of this study was to describe Se-induced modifications in the accumulation of elements important for the proper functioning of wheat, one of the most popular cereals. The presence of Se correlated with carbohydrate synthesis and electron paramagnetic resonance (EPR). This explained the mechanisms of Se's antioxidant activity. RESULTS: Selenium accumulation in vegetative and generative leaves, and in the grains of three wheat genotypes (cv. Parabola, cv. Raweta and cv. Manu), differing in their stress tolerance and grown hydroponically in the presence of 10 or 20 µM Na2 SeO4, , was proportional to its content in the medium. Stronger Se accumulation was typical of a stress-sensitive genotype. Selenium generally promoted the uptake of macronutrients and micronutrients but their distribution depended on tissue and genotype. Changes in the Se-induced EPR signals of paramagnetic metals and organic radicals corresponded with stress tolerance of the tested genotypes. CONCLUSIONS: Se application increased the accumulation of nutrients and carbohydrates that are vital for proper plant growth and development. Accelerated uptake of molybdenum (Mo), an element improving dietary properties of grains, may be an additional advantage of Se fertilization. The mechanisms of Se-induced changes in removing Mn and iron (Fe) ions from macromolecules may be one of the factors that differentiate plant tolerance to oxidative stress. © 2019 Society of Chemical Industry.

Changes in the physicochemical properties of barley and oat starches upon the use of environmentally friendly oxidation methods.

In this work, two environmentally friendly modification methods, UV irradiation and thermal treatment, were applied for the first time for modifications of oat and barley starches. Their impacts on starch properties were compared with those observed for starch oxidation with NaClO. XRD, EPR, FTIR, SEM and SEC methods were used to characterize the effects of modifications on starch structures. The decreases in molecular weight and crystallinity degree showed the destruction of starch structures upon prolonged UV irradiation and thermal treatment, more advanced in barley starch. The process of radical formation, studied by EPR, occurring to a larger extent in this starch, confirmed lower stability of barley starch structure. The alterations of starch structures correlated well with changes in its functional properties. It was found that UV irradiation was an effective oxidizing agent, whereas heating led mainly to the depolymerization of starch chains.

Properties of potato starch treated with microwave radiation and enriched with mineral additives.

The paper discusses how microwave radiation and mineral additives affect selected physical and chemical properties of potato starch. Potato starch was irradiated with microwaves of 440 W or 800 W and then enriched with ions of potassium, magnesium, copper, or iron. Atomic absorption spectroscopy showed an effective incorporation of ions, while mineral saturation depended on both the microwave radiation density and the element being introduced. The data obtained prove that the microwave radiation and saturation with minerals affect the rheology and thermal characteristics of the samples under examination. Both the degree and the vector of the transformation depend on the microwave output and the mineral component introduced. Generation of radicals depends on starch degradation at high temperature while microwaves affect the number of the radicals formed. Amounts of the heat-generated radicals depend on the microwave output and the type of metal ions introduced into the starch structure.

The effect of amylose content and level of oxidation on the structural changes of acetylated corn starch and generation of free radicals.

This study was aimed at determining the effect of starch oxidation on its acetylation, structure of starch granules, and generation of free radicals. Corn and waxy corn starches were oxidised by NaClO applied in doses of 10, 20, and 30g Cl/kg of starch, and then acetylated using acetic acid anhydride. The carboxyl, carbonyl, acetyl groups were determined in modified starches. Structural properties of starch granules were evaluated based on molecular weight distribution, gelatinisation, crystallinity, specific surface, intrinsic viscosity. EPR measurements were carried out to establish starch susceptibility to UV irradiation induced generation of free radicals. It was found that the number of carbon centered radicals was dependent on the kind of starch and its chemical modification. Study results allowed concluding that the applied modifications contributed to significant changes in starch granules that were determined not only by the amylose content of starch but also by the degree of its oxidation.

2D FTIR correlation spectroscopy and EPR analysis of Urtica dioica leaves from areas of different environmental pollution.

Leaves of Urtica dioica collected from two areas of different environmental pollution were analysed by fourier transform infrared spectroscopy (FTIR) and electron paramagnetic resonance (EPR) spectroscopy. Analysis of FTIR spectra allows to describe main component of plant like proteins, lipids and carbohydrates. Although the FTIR spectra of plants from these two geographical locations of different environmental pollution appear to be relatively similar, 2D correlation shows completely different patterns. Synchronous and asynchronous correlation maps showed sequences of changes occurring during development of plant, manly in Amide I and Amide II, lignin, lipids and cellulose. In addition, 2D analysis revealed another sequence of changes as the function of plant growth depending on the degree of the environmental pollution. Two various kinds of paramagnetic species, transition metal ions (Mn(II), Fe(III)) and stable organic radicals (chlorophyll, semiquinone, tyrosyl and carbon centered) were found in leaves of nettle collected at different stages of development and growing in clean and polluted environment. In plants growing in polluted area the injuries of protein molecules bonding metal ions and the disturbances of photosynthesis and redox equilibrium in cells, as well as instability of polysaccharide structure of cell walls were observed.

Influence of amylose content and oxidation level of potato starch on acetylation, granule structure and radicals' formation.

This study was aimed at determining the effect of the amylose content of starch and oxidation level of potato starch on the structure of starch granules, and susceptibility to chemical modification (acetylation) and subsequent generation of radicals. Potato starch and waxy potato starch were oxidised with sodium hypochlorite applied in doses corresponding to 10, 20, and 30gCl/kg starch, and then acetylated with acetic acid anhydride. The carboxyl, carbonyl, acetyl groups were determined in modified starches. Structural properties of starch granules were evaluated based on gelatinisation, crystallinity, specific surface, intrinsic viscosity, and microphotographs by SEM microscope. The electron paramagnetic resonance (EPR) measurements were carried out to establish starch susceptibility to radical creation upon chemical modification and UV radiation. The amount of formed radicals was treated as a measure of the starch structure stability. The higher amount of amylose and the highest level of oxidation led to strong starch structure destruction and consequently facilitated radical generation. Study results showed also that amylose content as well as the degree of starch oxidation modified consecutive acetylation process. The different effectiveness of the acetylation processes influenced the morphology and structure of starch granules.

The impact of short-term UV irradiation on grains of sensitive and tolerant cereal genotypes studied by EPR.

BACKGROUND: UV irradiation has ionisation character and leads to the generation of reactive oxygen species (ROS). The destructive character of ROS was observed among others during interaction of cereal grains with ozone and was caused by changes in structures of biomolecules leading to the formation of stable organic radicals. That effect was more evident for stress sensitive genotypes. In this study we investigated the influence of UV irradiation on cereal grains originating from genotypes with different tolerance to oxidative stress. RESULTS: Grains and their parts (endosperm, embryo and seed coat) of barley, wheat and oat were subjected to short-term UV irradiation. It was found that UV caused the appearance of various kinds of reactive species (O2-• , H2 O2 ) and stable radicals (semiquinone, phenoxyl and carbon-centred). Simultaneously, lipid peroxidation occurred and the organic structure of Mn(II) and Fe(III) complexes become disturbed. CONCLUSIONS: UV irradiation causes damage of main biochemical structures of plant tissues, the effect is more significant in sensitive genotypes. In comparison with ozone treatment, UV irradiation leads to stronger destruction of biomolecules in grains and their parts. It is caused by the high energy of UV light, facilitating easier breakage of molecular bonds in biochemical compounds. © 2017 Society of Chemical Industry.

Electron Paramagnetic Resonance (EPR) Spectroscopy in Studies of the Protective Effects of 24-Epibrasinoide and Selenium against Zearalenone-Stimulation of the Oxidative Stress in Germinating Grains of Wheat.

These studies concentrate on the possibility of using selenium ions and/or 24-epibrassinolide at non-toxic levels as protectors of wheat plants against zearalenone, which is a common and widespread mycotoxin. Analysis using the UHPLC-MS technique allowed for identification of grains having the stress-tolerant and stress-sensitive wheat genotype. When germinating in the presence of 30 µM of zearalenone, this mycotoxin can accumulate in both grains and hypocotyls germinating from these grains. Selenium ions (10 µM) and 24-epibrassinolide (0.1 µM) introduced together with zearalenone decreased the uptake of zearalenone from about 295 to 200 ng/g and from about 350 to 300 ng/g in the grains of tolerant and sensitive genotypes, respectively. As a consequence, this also resulted in a reduction in the uptake of zearalenone from about 100 to 80 ng/g and from about 155 to 128 ng/g in the hypocotyls from the germinated grains of tolerant and sensitive wheat, respectively. In the mechanism of protection against the zearalenone-induced oxidative stress, the antioxidative enzymes-mainly superoxide dismutase (SOD) and catalase (CAT)-were engaged, especially in the sensitive genotype. Electron paramagnetic resonance (EPR) studies allowed for a description of the chemical character of the long-lived organic radicals formed in biomolecular structures which are able to stabilize electrons released from reactive oxygen species as well as the changes in the status of transition paramagnetic metal ions. The presence of zearalenone drastically decreased the amount of paramagnetic metal ions-mainly Mn(II) and Fe(III)-bonded in the organic matrix. This effect was particularly found in the sensitive genotype, in which these species were found at a smaller level. The protective effect of selenium ions and 24-epibrassinolide originated from their ability to inhibit the destruction of biomolecules by reactive oxygen species. An increased ability to defend biomolecules against zearalenone action was observed for 24-epibrassinolide.

Structural and biochemical response of chloroplasts in tolerant and sensitive barley genotypes to drought stress.

The aim of this research was to characterize the changes of structural organization of chloroplasts of sensitive (Maresi) and tolerant (Cam/B1) barley genotypes upon soil drought (10days), which was applied in two stages of plant growth, i.e. seedlings and flag leaves. The electron paramagnetic resonance (EPR) technique was used for the determination of changes in the concentration and nature of long-lived radicals and metal ions (Mn, Fe), measured directly in the structures of fresh leaves, occurring after stress treatment. Stronger variations of EPR parameters were found after drought stress application in the flag-leaf phase and for sensitive genotype. Chloroplasts of Cam/B1 were characterized by a larger surface area and less degradation of their structure during drought stress in comparison to Maresi. The data obtained from Raman spectra showed that better stress tolerance of the genotype was accompanied by greater accumulation of carotenoids in chloroplasts and was correlated with an increase in carotenoid radicals. The increase of the value of the electrokinetic potential (relative to control), which was slightly larger for the chloroplasts of Maresi than of Cam/B1, indicated the chemical reconstruction of the membrane leading to a reduction of their polarity during drought action.

The impact of biochemical composition and nature of paramagnetic species in grains on stress tolerance of oat cultivars.

The aim of this work was to investigate the relationships between the chemical composition of oat grains and the tolerance to oxidative stress of oat genotypes. The studies were based on the results of biochemical analyses and both EPR and Raman spectroscopies on whole grains and their parts (embryo, endosperm, seed coat) originating from oat genotypes with different sensitivities to stress. We found that the amounts of fats and especially unsaturated fatty acids, proteins rich in glutamic acid and glycine, as well as phenolics and tocopherols were higher in grains of the tolerant genotype. Moreover, fats and proteins were distributed not only in embryos, but also in endosperms. The grains of tolerant genotypes exhibited high antioxidant activity and contained greater amounts of ß-glucan. EPR data pointed to higher concentrations of various kinds of stable organic radicals (semiquinone, tyrosyl and carbon-centered radicals) in whole grains (and their parts) of sensitive genotypes. EPR spectra revealed the character of interactions of paramagnetic transition metal ions Fe(III) and Mn(II) with organic and inorganic structures of grains. The quantitative EPR measurements showed the dependence between the amount of radical species and the content of transition metal ions, mainly Fe(III) bonded to inorganic structures.

Vibration-reorientation dynamics, structural changes and its interrelation with the phase transition in polycrystalline [Cr(OC(NH2)2)6](BF4)3.

Polycrystalline hexakis(urea-O)chromium(III) tetrafluoroborate possesses in the temperature range of 295-105 K one solid-solid phase transition at T(C) ≈ 255 K. Analysis of the band shapes associated with the ν(as)(CN) and δ(as)(NH2) vibrational modes of the Fourier transform infrared absorption (FT-IR) spectra, registered in the temperature range of 295-10 K, indicated existence of fast (τ(R) ≈ 10(-12) s) reorientational motion of the protons from NH2 groups belonging to OC(NH2)2 (urea) ligands, which does not suddenly change at T(C). Moreover, splitting of the IR bands associated with the ν(as)(NH), ν(s)(NH) and ν(as)(BF)F2/ν(s)(CN) modes at T(C) indicated that this phase transition is associated with a change of crystal structure. Similar analysis of the Raman scattering bands (FT-RS), associated with the δ(s)(FBF)E, ν(s)(BF)A and ν(as)(BF)F2/ν(s)(CN) vibrational modes, indicated fast reorientation of the BF4(−) ions, which does not suddenly change at T(C), and additionally confirmed structural character of this phase transition. Results obtained from vibrational spectroscopy measurements are compatible with that obtained by electron paramagnetic resonance spectroscopy (EPR) measurements in function of temperature, where rapid narrowing of the EPR line in the vicinity of the T(C) was observed.

Changes of paramagnetic species in cereal grains upon short-term ozone action as a marker of oxidative stress tolerance.

The increase of the concentration of ozone in the atmosphere, being the direct source of reactive oxygen species, results in the yield loss of agronomic crops. On the other hand, ozone is also used as a protector against microorganisms, living in plants and present in materials obtained from them, dangerous for human and animal health. In this work it has been studied if ozone in doses similar to those used for removal of microorganisms can have significant influence on the generation of stable organic radicals and changes in the character of transition metal ions and in the antioxidative biochemical parameters of cereal grains. The aim of this work was to find if the response of grains of three cereals (wheat, oat and barley) to ozone depended on their oxidative stress tolerance. The influence of direct short-term ozone application on grains of these cereals, each represented by two genotypes with different oxidative stress tolerance, was studied by biochemical analyses and by electron paramagnetic resonance (EPR) technique. Whole grains as well as their parts: embryo, endosperm and seed coat were subjected to ozone treatment for 30 min. Biochemical investigation of control samples showed that their antioxidant activity increased in order: wheat

Stable radicals and biochemical compounds in embryos and endosperm of wheat grains differentiating sensitive and tolerant genotypes--EPR and Raman studies.

The aim of this study was to uncover the specific species in grains that might differentiate the wheat genotypes according to their tolerance to oxidative stress. Measurements by EPR and Raman spectroscopy techniques were used to examine whole grains and their parts (embryo, endosperm, seed coat) originating from four wheat genotypes with differing tolerance to drought stress. Raman spectra showed that, in spite of the similar amounts of proteins in whole grains from tolerant and sensitive genotypes, in tolerant ones they were accumulated mainly in embryos. Moreover, in embryos from these grains, a higher content of unsaturated fatty acids was observed. Endosperm of grains from the tolerant genotype, richer with starch than that of sensitive one, exhibited higher content of amylopectin. Detailed analysis of EPR signals and simulation procedures of the spectra allowed the estimation of the nature of interactions of Fe(III) and Mn(II) with organic and inorganic structures of grains and the character of organic stable radicals. Three types of these radicals: carbohydrate, semiquinone and phenoxyl, were identified. The amounts of these radicals were higher in grains of sensitive genotypes, mostly because of differences in carbohydrate radical content in endosperm. Taking into account the level of radical concentration and greater capacity for radical formation in grains from plants of lower tolerance to stress, the content of radicals, especially of a carbohydrate nature, was considered as a marker of the plant resistance to stress conditions.

The influence of the starch component on thermal radical generation in flours.

Transition metal ions and radicals in flours of various botanical origins with different content of starch have been studied by EPR before and after thermal treatment. The amounts of metal ions, have been determined by ICP OES. Simulations of EPR spectra have revealed the presence of several types of radicals (carbon-centred, tyrosyl and semiquinone) localized in starch and protein fractions of flours. Thermal treatment of flours significantly increased the amount of radicals with a simultaneous decrease of the signal intensity of transition metal ions. The proposed mechanism of thermal generation of stable organic radicals was associated with the redox processes involving transition metal ions, which facilitated the formation of radicals. The dependence between the way starch is treated and the mechanism of radical formation was also shown.

Effect of mineral elements on physicochemical properties of oxidised starches and generation of free radicals.

The objective of this study was to determine the effect of enrichment of oxidised starches with mineral compounds on their physicochemical properties and capability for free radical generation. Potato and spelt wheat starches were oxidised with sodium hypochlorite and, afterwards, modified with ions of potassium, magnesium and iron. The modified starches were analysed for: content of mineral elements, colour parameters (L*a*b*), water binding capacity solubility in water at temperature of 50 and 80 °C, and susceptibility to enzymatic hydrolysis with α-amylase. In addition, thermodynamic characteristics of gelatinisation was determined by differential scanning calorimetry (DSC), and the number and character of thermally generated free radicals was assayed using electron paramagnetic resonance (EPR). Based on the results achieved, it was concluded that the quantity of incorporated minerals and changes in the assayed physicochemical parameters depended not only on the botanical type of starch but also on the type of the incorporated mineral element.

The mechanism of thermal activated radical formation in potato starch studied by electron paramagnetic resonance and Raman spectroscopies.

Degradation of starch and the constituent fractions: amylopectin and amylose during thermal treatment in the range 423-503 K was investigated by electron paramagnetic resonance (EPR) and Raman spectroscopy. Degradation process was accompanied by the generation of carbon-centered stable radicals. EPR provided data on the nature and structure of radicals and on their evolution upon thermal treatment, whereas Raman spectroscopy allowed monitoring the changes of bonds in polysaccharides. It was found that amylose was the most susceptible toward high temperatures and the process of radical generation started at lower temperature than in amylopectin and starch, which were more resistant to thermal degradation.