Temperature-Caused Changes in Raman Pattern and Protein Profiles of Winter Triticale (x Triticosecale, Wittm.) Field-Grown Seedlings.

Climate change, which causes periods with relatively high temperatures in winter in Poland, can lead to a shortening or interruption of the cold hardening of crops. Previous research indicates that cold acclimation is of key importance in the process of acquiring cereal tolerance to stress factors. The objective of this work was to verify the hypothesis that both natural temperature fluctuations and the plant genotype influence the content of metabolites as well as proteins, including antioxidant enzymes and photosystem proteins. The research material involved four winter triticale genotypes, differing in their tolerance to stress under controlled conditions. The values of chlorophyll a fluorescence parameters and antioxidant activity were measured in their seedlings. Subsequently, the contribution of selected proteins was verified using specific antibodies. In parallel, the profiling of the contents of chlorophylls, carotenoids, phenolic compounds, and proteins was carried out by Raman spectroscopy. The obtained results indicate that a better PSII performance along with a higher photosystem II proteins content and thioredoxin reductase abundance were accompanied by a higher antioxidant activity in the field-grown triticale seedlings. The Raman studies showed that the cold hardening led to a variation in photosynthetic dyes and an increase in the phenolic to carotenoids ratio in all DH lines.

Raman spectroscopy of lymphocytes from patients with the Epstein-Barr virus infection.

In this study, Raman spectroscopy is applied to trace lymphocytes activation following contact with the Epstein-Barr virus (EBV) of the herpesvirus family. The biomarker of cell activation is found to be the 520 cm-1 band, indicating formation of immunoglobulins. The blood samples are obtained from patients diagnosed with infectious mononucleosis and treated at the University Hospital in Kraków. The lymphocytes' Raman spectra are collected using a mapping technique, exciting samples with a 514.5 nm line of Ar + laser. Measurements are performed on the 1st, 4th, 6th, 12th and 30th day of hospitalization, until the patient has recovered. The highest intensity of the immunoglobulin marker is observed on the 4th day of hospitalization, while the results of the blood count of patients show the greatest increase in the number of lymphocytes at the beginning of hospitalization. No activated lymphocytes were observed in the blood of healthy volunteers. Some information is provided by the evaluation of B-cell activation by estimating the activated areas in the cells, which are determined by the presence of the Ig marker. The 900 cm-1 band and band around 1450 cm-1 are also analyzed as markers of the presence of the latent membrane protein, LMP2A (and 2B), of the EBV viral protein. The anomalous degree of depolarization observed in B-cells in the course of EBV infection appears to be due to the influence of a virus protein, disrupting BCR signal transduction.

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.

Influence of the type of substrate on the properties of carbon nanotubes layer studied by Raman spectroscopy.

The development of nanomaterials technology allows to design a novel medical strategies, and could also be useful in the field of regenerative medicine. The paper presents a study on the functionalized multi-walled carbon nanotubes (MWCNTs-f) layers deposited by electrophoretic method (EPD) on the surfaces of two types of substrates: titanium (Ti) and stainless steel. SEM and EDS analyses confirm that incubation in a simulated body fluid (SBF) caused a formation of hydroxyapatite on the surface of the Ti/MWCNTs-f. Raman micro-spectroscopy was a method of choice to study presented materials. The MWCNTs-f layer on the surface of the titanium plate shows better layer order than the corresponding layer deposited on the stainless steel. The structure and ordering of the nanocarbon layer play a key role in the biological activity of the materials. This was confirmed by the incubation of the plates with deposited layer of carbon nanotubes in SBF. A titanium substrate with a MWCNTs-f layer supports the deposition of some components from the environment, while a stainless steel substrate promotes the formation of a carbon film that inhibits the deposition of certain components from the environment. A two-trace two-dimensional (2T2D) analysis confirmed a different effect of SBF on the MWCNTs-f layer depending on the type of substrate. The MWCNTs-f layer on titanium substrate seems to represent an interesting proposition for novel bioactive strategies.

Air pollution in the places of Betula pendula growth and development changes the physicochemical properties and the main allergen content of its pollen.

Pollen allergy becomes an increasing problem for humans, especially in the regions, where the air pollution level increases due to the traffic and urbanization. These factors may also affect the physiological activity of plants, causing changes in pollen allergenicity. The aim of the study was to estimate the influence of air pollutants on the chemical composition of birch pollen and the secondary structures of the Bet v1 protein. The research was conducted in seven locations in Malopolska region, South of Poland of a different pollution level. We have found slight fluctuations in the values of parameters describing the photosynthetic light reactions, similar spectra of leaf reflectance and the negligible differences in the discrimination values of the δ13C carbon isotope were found. The obtained results show a minor effect of a degree of pollution on the physiological condition B. pendula specimen. On the other hand, mean Bet v1 concentration measured in pollen samples collected in Kraków was significantly higher than in less polluted places (p = .03886), while FT-Raman spectra showed the most distinct variations in the wavenumbers characteristic of proteins. Pollen collected at sites of the increased NOx and PM concentration, show the highest percentage values of potential aggregated forms and antiparallel ß-sheets in the expense of α-helix, presenting a substantial impact on chemical compounds of pollen, Bet v1 concentration and on formation of the secondary structure of proteins, what can influence their functions.

A study of the interactions between human osteoblast-like cells and polymer composites with functionalized graphene derivatives using 2D correlation spectroscopy (2D-COS).

In response to the growing need for development of modern biomaterials for applications in regenerative medicine strategies, the research presented here investigated the biological potential of two types of polymer nanocomposites. Graphene oxide (GO) and partially reduced graphene oxide (rGO) were incorporated into a poly(ε-caprolactone) (PCL) matrix, creating PCL/GO and PCL/rGO nanocomposites in the form of membranes. Proliferation of osteoblast-like cells (human U-2 OS cell line) on the surface of the studied materials confirmed their biological activity. Fluorescence microscopy was able to distinguish the different patterns of interaction between cells (depending on the type of material) after 15 days of the test run. Raman micro-spectroscopy and two-dimensional correlation spectroscopy (2D-COS) applied to Raman spectra distinguished the nature of cell-material interactions after only 8 days. Combination of these two techniques (Raman micro-spectroscopy and 2D-COS analysis) facilitated identification of a much more complex cellular response (especially from proteins) on the surface of PCL/GO. The presented approach can be regarded as a method for early study of the bioactivity of membrane materials.

Blood-stage antiplasmodial activity and oocyst formation-blockage of metallo copper-cinchonine complex.

In the fight against malaria, the key is early treatment with antimalarial chemotherapy, such as artemisinin-based combination treatments (ACTs). However, Plasmodium has acquired multidrug resistance, including the emergence of P. falciparum strains with resistance to ACT. The development of novel antimalarial molecules, that are capable of interfering in the asexual and sexual blood stages, is important to slow down the transmission in endemic areas. In this work, we studied the ability of the mettalo copper-cinchonine complex to interfere in the sexual and asexual stages of Plasmodium. The tested compound in the in vitro assay was a cinchonine derivative, named CinCu (Bis[Cinchoninium Tetrachlorocuprate(II)]trihydrate). Its biological functions were assessed by antiplasmodial activity in vitro against chloroquine-resistant P. falciparum W2 strain. The mice model of P. berghei ANKA infection was used to analyze the antimalarial activity of CinCu and chloroquine and their acute toxicity. The oocyst formation-blocking assay was performed by experimental infection of Anopheles aquasalis with P. vivax infected blood, which was treated with different concentrations of CinCu, cinchonine, and primaquine. We found that CinCu was able to suppress as high as 81.58% of parasitemia in vitro, being considered a molecule with high antiplasmodial activity and low toxicity. The in vivo analysis showed that CinCu suppressed parasitemia at 34% up to 87.19%, being a partially active molecule against the blood-stage forms of P. berghei ANKA, without inducing severe clinical signs in the treated groups. The transmission-blocking assay revealed that both cinchonine and primaquine were able to reduce the infection intensity of P. vivax in A. aquasalis, leading to a decrease in the number of oocysts recovered from the mosquitoes' midgut. Regarding the effect of CinCu, the copper-complex was not able to induce inhibition of P. vivax infection; however, it was able to induce an important reduction in the intensity of oocyst formation by about 2.4 times. It is plausible that the metallo-compound also be able to interfere with the differentiation of parasite stages and/or ookinete-secreted chitinase into the peritrophic matrix of mosquitoes, promoting a reduction in the number of oocysts formed. Taken together, the results suggest that this compound is promising as a prototype for the development of new antimalarial drugs. Furthermore, our study can draw a new pathway for repositioning already-known antimalarial drugs by editing their chemical structure to improve the antimalarial activity against the asexual and sexual stages of the parasite.

Does Nitrogen Fertilization Affect the Secondary Structures of Gliadin Proteins in Hypoallergenic Wheat?

One of the macronutrients indispensable for plant growth and development is nitrogen (N). It is responsible for starch and storage protein (gliadins and glutenins) biosynthesis and, in consequence, influences kernels' quality and yields. However, applying N-fertilizers increases gluten content in wheat, and it may intensify the risk of developing allergy symptoms in gluten-sensitive individuals. The purpose of our research was to analyse whether and how the elimination of N-fertilizers during the cultivation of wasko.gl- wheat (modified genotype lacking ω-gliadins) changes the secondary structures of gliadin proteins. To this aim, using the FT-Raman technique, we examined flour and gliadin protein extracts obtained from kernels of two winter wheat lines: wasko.gl+ (with a full set of gliadin proteins) and wasko.gl- (without ω-gliadin fraction) cultivated on two different N-fertilization levels-0 and 120 kg N·ha-1. On the basis of the obtained results, we proved that nitrogen fertilization does not have a major impact on the stability of the secondary structures of gliadin proteins for wasko.gl- wheat line with reduced allergenic properties. Furthermore, the results presented herein suggest the possibility of increasing the stability of glutenin structures as a result of the N-fertilization of wasko.gl- wheat line, which gives hope for its use in the production of wheat articles devoted to people suffering from diseases related to gluten sensitivity.

Application of 2D correlation analysis in FT - Raman investigations of quinidine aqueous solutions with varying pH.

An analysis of FT-Raman spectra of quinidine (C20H24N2O2) aqueous solutions with varying pH (which was regarded as an external perturbation) was performed using the 2D correlation method. The main course of changes in the quinidine solution appears to be: protonation changes of the quinuclidine nitrogen N1, followed by protonation changes of nitrogen N13 in the quinoline, leading to the appearance of cross-peaks in the synchronous and asynchronous correlation maps. The intensity changes of peaks at 1369 cm-1 for the unprotonated quinidine molecule, and characteristic peaks at 1387 cm-1 and 1389 cm-1 for protonated quinuclidine and double protonated quinidine, respectively, along with the decrease in pH, confirmed that the change in the pH of the quinidine solution has an influence on the protonation process of the Cinchona alkaloid. The negative synchronous and asynchronous cross-peaks at (1385, 823) cm-1 and (1387, 822) cm-1, respectively, indicate the importance of remodeling the quinoline fragment, during the process of a double protonation of the quinidine molecule. Bands correlating with 2809 cm-1 confirmed the importance of the methoxy group in the process of quinidine protonation. The creation of hydrogen bonds after double protonation of the Cinchona alkaloids, assisted by the CH3-O group, give an interesting insight into the changes in the studied compound occurring along with a decrease in pH.

Early Recognition of the PCL/Fibrous Carbon Nanocomposites Interaction with Osteoblast-like Cells by Raman Spectroscopy.

Poly(ε-caprolactone) (PCL) is a biocompatible resorbable material, but its use is limited due to the fact that it is characterized by the lack of cell adhesion to its surface. Various chemical and physical methods are described in the literature, as well as modifications with various nanoparticles aimed at giving it such surface properties that would positively affect cell adhesion. Nanomaterials, in the form of membranes, were obtained by the introduction of multi-walled carbon nanotubes (MWCNTs and functionalized nanotubes, MWCNTs-f) as well as electro-spun carbon nanofibers (ESCNFs, and functionalized nanofibers, ESCNFs-f) into a PCL matrix. Their properties were compared with that of reference, unmodified PCL membrane. Human osteoblast-like cell line, U-2 OS (expressing green fluorescent protein, GFP) was seeded on the evaluated nanomaterial membranes at relatively low confluency and cultured in the standard cell culture conditions. The attachment and the growth of the cell populations on the polymer and nanocomposite samples were monitored throughout the first week of culture with fluorescence microscopy. Simultaneously, Raman microspectroscopy was also used to track the dependence of U-2 OS cell development on the type of nanomaterial, and it has proven to be the best method for the early detection of nanomaterial/cell interactions. The differentiation of interactions depending on the type of nanoadditive is indicated by the ν(COC) vibration range, which indicates the interaction with PCL membranes with carbon nanotubes, while it is irrelevant for PCL with carbon nanofibers, for which no changes are observed. The vibration range ω(CH2) indicates the interaction for PCL with carbon nanofibers with seeded cells. The crystallinity of the area ν(C=O) increases for PCL/MWCNTs and for PCL/MWCNTs-f, while it decreases for PCL/ESCNFs and for PCL/ESCNFs-f with seeded cells. The crystallinity of the membranes, which is determined by Raman microspectroscopy, allows for the assessment of polymer structure changes and their degradability caused by the secretion of cell products into the ECM and the differentiation of interactions depending on the carbon nanostructure. The obtained nanocomposite membranes are promising bioactive materials.

FT-Raman Spectroscopy as a Tool to Study the Secondary Structures of Wheat Gliadin Proteins.

Raman spectroscopy is a useful method in biological, biomedical, food, and agricultural studies, allowing the simultaneous examination of various chemical compounds and evaluation of molecular changes occurring in tested objects. The purpose of our research was to explain how the elimination of ω-fractions from the wheat gliadin complex influences the secondary structures of the remaining αßγ-gliadins. To this aim, we analyzed the endosperm of wheat kernels as well as gliadin proteins extracted from two winter wheat genotypes: wasko.gl+ (control genotype containing the full set of gliadins) and wasko.gl- (modified genotype lacking all ω-gliadins). Based on the decomposition of the amide I band, we observed a moderate increase in ß-forms (sheets and turns) at the expense of α-helical and random coil structures for gliadins isolated from the flour of the wasko.gl- line. Since ω-gliadins contain no cysteine residues, they do not participate in the formation of the disulfide bridges that stabilize the protein structure. However, they can interact with other proteins via weak, low-energetic hydrogen bonds. We conclude that the elimination of ω-fractions from the gliadin complex causes minor modifications in secondary structures of the remaining gliadin proteins. In our opinion, these small, structural changes of proteins may lead to alterations in gliadin allergenicity.

Origin and occurrence of gem-quality, skarn-hosted barite from Jebel Ouichane near Nador in Morocco.

Light-blue barite from Jebel Ouichane in Morocco forms blade-like tabular crystals (up to ca. 10 cm) with superb transparency and lustre and represents one of the most spectacular gem-quality worldwide. The barite is hosted by iron-ore-bearing skarns, developed within Jurassic-Cretaceous limestones, and occurs in close spatial association with calcite. The crystals have their cores enriched in Sr and contain abundant monophase (liquid) fluid inclusions of primary and pseudosecondary origin. The barite probably precipitated slowly at a relatively low supersaturation and under the control of a surface reaction precipitation mechanism. However, there were some episodes during its formation with a fast growth rate and the coupled dissolution and recrystallization processes. A combination of fluid inclusion data and stable δ18O value for barite (+ 6.71‰ VSMOW) suggests that low-salinity barite-forming solutions resulted from the mixing of strongly-diluted meteoric waters (enriched in light oxygen isotope) with magmatic-hydrothermal fluids under low-temperature conditions (< 100 °C). Meanwhile, the mineralizing fluids must have been enriched in Ba, Sr, Ca, Mg, and other elements derived from the alteration of carbonate and silicate minerals in sedimentary and igneous rocks. The coupling between sulphur and oxygen isotope data (+ 16.39‰ VCDT and + 6.71‰ VSMOW, respectively) further suggests that barite crystallized in steam-heated environment, where SO42- derived from magmatic-hydrothermal SO2 reacted with sulphates that originate from the oxidation of H2S under near-surface conditions.

Raman studies of the interactions of fibrous carbon nanomaterials with albumin.

Adsorption or immobilization of proteins on synthetic surfaces is a key issue in the context of the biocompatibility of implant materials, especially those intended for the needs of cardiac surgery but also for the construction of biosensors or nanomaterials used as drug carriers. The subject of research was the analysis of Raman spectra of two types of fibrous carbon nanomaterials, of great potential for biomedical applications, incubated with human serum albumin (HSA). The first nanomaterial has been created on the layer of MWCNTs deposited by electrophoretic method (EPD) and then covered by thin film of pyrolytic carbon introduced by chemical vapor deposition process (CVD). The second material was formed from carbonized nanofibers prepared via electrospinning (ESCNFs) of polyacrylonitrile (PAN) precursor and then covered with pyrolytic carbon (CVD). The G-band blue-shift towards the position of about 1600cm-1, observed for both studied surfaces, clearly indicates the albumin (HSA) adhesion to the surface. The G and G' (2D) peak shift was employed to assess the stress build up on the carbon nanomaterials. The surface nano- and micro-topography as well as the method of ordering the carbon nanomaterial has a significant influence on the mode of surface-protein interaction.

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.

Epileptic rat brain tissue analyzed by 2D correlation Raman spectroscopy.

Absence epilepsy is the neurological disorder characterized by the pathological spike-and wave discharges present in the electroencephalogram, accompanying a sudden loss of consciousness. Experiments were performed on brain slices obtained from young male WAG/Rij rats (2-3weeks old), so that they were sampled before the appearance of brain-damaging seizures symptoms. Two differing brain areas of the rats' brain tissue were studied: the somatosensory cortex (Sc) and the dorsal lateral geniculate nucleus of the thalamus (DLG). The Raman spectra of the fresh brain scraps, kept during measurements in artificial cerebrospinal fluid, were collected using as an excitation source 442nm, 514.5nm, 785nm and 1064nm laser line. The average spectra were analyzed by 2D correlation method regarding laser line as an external perturbation. In 2D synchronous spectra positive auto-peaks corresponding to the CC stretching and amide I band vibrations show maxima at 1660cm-1 and 1662cm-1 for Sc and DLG, respectively. The prominent auto-peak at 2937cm-1, originated from the CH3 mode in DLG brain area, seems to indicate the importance of methylation, considered to be significant in epileptogenesis. Synchronous and asynchronous correlations peaks, glutamic acid and gamma-aminobutyric acid (GABA), appear in Sc and DLG, respectively. In the 1730-1600cm-1 range occur cross-peaks which appearance might be triggered by glial fibrillary acidic protein (GFAP) activation.

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.

The ring-stage of Plasmodium falciparum observed in RBCs of hospitalized malaria patients.

Raman spectra of the blood samples obtained directly from hospitalized malaria patients with Plasmodium falciparum (P. falciparum) in the ring-stage were analyzed. Changes observed in the Raman band intensities of the infected patients compared to healthy volunteers are the result of parasite activity inside red blood cells. The obtained spectra were discussed by analyzing differences in particular spectral regions by evaluating changes in the band intensity ratios as well as using PCA analysis. The alterations of erythrocyte membranes caused by parasite penetration are visible by a reduced I1130/I1075 intensity ratio expressing the lowering of the amount of domains arranged in trans conformation. The I2930/I2850 ratio, which is a measure of modifications in structures of membrane proteins and lipids, in infected red blood cells increases, which is caused by malaria protein export to the erythrocyte membrane and expresses the membrane disarrangement. In the pyrrole ring vibration region, the ν4 band marker of the oxygenated-Hb shows at 1371 cm(-1) whereas the ν4 band at 1353 cm(-1) related to the deoxygenated-Hb is observed for malaria patients and is characterized by a higher intensity in infected erythrocytes. The amide I analysis shows the modifications in the secondary structure composition in the infected RBCs. We found that the P. falciparum infection leads to a decrease in the α-helical content and a concurrent increase in undefined (random-coil) structures. It was observed that the Raman spectra changes are also the result of the hemozoin formation process. In the pyrrole ring stretching vibration region, the increase of 1220 cm(-1) (deoxyHb) as against 1248 cm(-1) (oxyHb) may be considered as a signal of hemozoin formation in the RBCs. Relatively intense band patterns at 1560 cm(-1) and also at 1570 cm(-1) and 1552 cm(-1) may be due to the hemozoin that is formed according to parasite activity. The results of medical diagnostic tests had not presented changes in patient RBC parameters. A significant reduction in WBC count was noticed along with a decrease in neutrophil and platelet count when compared with the control group. Although no change is observed in the overall picture of the erythrocytes, pathological changes are evident in the Raman spectrum.

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.

Micas from mariupolite of the Oktiabrski massif (SE Ukraine): an insight into the host rock evolution--geochemical data supported by Raman microspectroscopy.

Muscovite and two dark mica varieties (the coarse-crystalline, pegmatitic, and fine-crystalline with signs of early weathering) representing members of the biotite series, originating from mariupolite of the Oktiabrski massif, (Ukraine), were investigated along with their solid inclusions using electron microprobe and Raman micro-spectroscopy to discuss their genesis and relationship to the parental magma. The coarse-crystalline, pegmatitic biotite, (K1.90Rb0.02Na0.01)(Fe3.56(2+)Mg1.34Ti0.36Fe0.34(3+)Mn0.03)[(Si5.73Al2.10Fe0.17(3+))O20](OH3.24 F0.76) represents the primary, magmatic annite that crystallized from an alkaline, Fe-rich and Mg-depleted host magma, whereas the fine-crystalline biotite, partly altered to vermiculite, (K1.75Rb0.03Na0.03)(Fe3.23(3+)Fe1.16(2+)Mg0.26Mn0.04Ti0.10)[(Si5.16 Al2.84)O20](OH)4.00, devoid of F, represents a re-equilibrated or secondary, post-magmatic Fe(3+)-bearing mica crystallized from alkaline to the subalkaline host magma. Muscovite, (K1.96Na0.06)(Al3.97Fe0.06(2+))[(Si5.99Al2.01)O20](OH)4, with low Na/(Na+K) ratio, low Fe and devoid of Ti and also F, forms only tiny, subhedral flakes formed in the post-magmatic, hydrothermal stage. The primary, unaltered biotite contains numerous solid inclusions of primary origin (albite, aegirine, zircon, K-feldspar, nepheline, pyrochlore, magnetite) and secondary origin (natrolite, hematite, Ti-Mn oxides/hydroxides); most of them are accompanied by a carbonaceous substance, all confirmed by scanning electron microscopy and Raman microspectroscopy.