National Synchrotron Light Laboratory Research Articles

EVALUATION OF REUSE WATER FROM THE PINHEIROS METRO STATION IN THE CITY OF SÃO PAULO (BRAZIL) FOR THE PRESENCE OF METALS USING THE X-RAY FLUORESCENCE TECHNIQUE WITH SYNCHROTRON RADIATION

Ensuring the conservation of water reserves is a challenge for all humanity, which makes it essential to continuously study new ways of treating and using water sources on the planet. One of the focuses of water conservation and sustainability is the reuse of water sources in order to guarantee the continuity of the main reserves of drinking water for activities such as waterlogging and food production. The present study aimed to investigate the quality of the water used for reuse at the Pinheiros subway station in the city of São Paulo (Brazil) in order to assess the degree of presence of heavy metals and other chemical elements using the X-ray fluorescence technique by synchrotron radiation at the National Synchrotron Light Laboratory (LNLS). The data collection for a sample collected in the summer and analyzed revealed a pH of 6.8 and the presence of 19 chemical elements using the technique with concentrations in the order of ng.mL -1 , with emphasis on the presence of lead (Pb).

Elemental distribution by synchrotron X‐ray microfluorescence of prostate 3D cell culture

Prostate cancer is a highly prevalent disease and ranks second among malignant neoplasms that affect men around the world, behind lung cancer alone. Trace elements are very important and are involved in many cellular processes. The X‐ray microfluorescence technique is an advanced tool of high spatial resolution, sensitivity, multielemental analysis, and nondestructiveness for trace element study. This study aimed to investigate the elemental distribution in spheroids obtained through the following human prostate cell lines using synchrotron X‐ray microfluorescence: tumor cell line androgen independent (DU145), tumor cell line androgen dependent (LNCaP), and normal cell line (RWPE‐1). The measurements were performed with a standard geometry of 45° of incidence, excited by a white beam using a pixel of 25 μm and an acquisition time of 300 ms/pixel at the X‐ray fluorescence beamline at the Synchrotron Light National Laboratory (Campinas, Brazil). The synchrotron X‐ray microfluorescence results showed differences between groups in all elements analyzed and suggested that further studies should be performed to understand the relationship of these trace elements with the progression and development of the disease.

Nanomateriais à base de óxidos de ferro suportados com resíduos ricos em sílica

As Nanopartículas podem ser obtidas através de diferentes componentes químicos, como por exemplo, os óxidos de ferro que apresentam como vantagens a facilidade de síntese, baixa toxicidade, biocompatibilidade, alta capacidade de adsorção e baixo custo. Uma vez que, as NPs possuem características bastante peculiares diferindo significativamente de outros materiais principalmente pelo aumento da área de superfície e também pelos efeitos quânticos. O trabalho tem o objetivo de obter Nanomateriais à base de óxidos de ferro suportados em diferentes resíduos (cinza de casca de arroz, cerâmica vermelha e ágata), compostos majoritariamente por SiO2, através do método poliol, e avaliar a possibilidade do uso destes em diferentes aplicações, como por exemplo, na área da catálise e em diversos processos da engenharia. Os Nanomateriais foram caracterizados com técnicas experimentais existentes no Laboratório Nacional de Luz Síncrotron (LNLS), que permitem a investigação eletrônica e estrutural dos Nanomateriais como a Espectroscopia de Fotoelétrons Induzidos por Raios X (XPS). O estudo permitiu identificar que foi possível o uso de resíduos com SiO2 na obtenção de NPs, bem como, confirmou que há diferenciação na estruturação destas. Possibilitando a expansão dos estudos com relação à aplicação destes Nanomateriais.

Cerebrospinal fluid analysis of patients with als by x-ray microfluorescence

Introduction: Amyotrophic Lateral Sclerosis (ALS) is the most common motor neuron disease in adults but its etiology remains unknown. Clinically, the patient manifests progressive weakness, paralysis and, usually, evolves to death between 2 to 5 years. The analysis of biological samples provides a powerful strategy to investigate pathological processes and studies indicate an important role of trace elements in ALS. Objective: In this work we aimed to investigate the levels of trace elements in CSF samples from patients with ALS. Methodology: CSF samples from cases of ALS (10) and control cases (6) were treated for analysis of trace elements by X-ray Microfluorescence with Synchrotron Radiation. The measurements were performed on the XRF beam line at the National Synchrotron Light Laboratory (Campinas, Brazil). The intensity of the X-ray spectrum for each element was obtained with AXIL software (distributed by the International Atomic Energy Agency). The data obtained were statistically analyzed by General Linear Models, in a method based on multivariate analysis. Results and Discussion: In this study, the following elements were analyzed: aluminum, bromine, calcium, chlorine, copper, chromium, iron, potassium, phosphorus, nickel, rubidium, silicon, sulfur and zinc. In CSF samples from ALS cases a significant increase of calcium, chlorine and potassium concentration was observed when compared to samples from control cases. Conclusion: Excitotoxicity is one of the many factors implicated in the pathogenic process of ALS. The intracellular calcium influx appears to contribute to neurodegeneration in multiple pathways, leading to cell death. Activation of potassium and chlorine channels by increased calcium level has been described and prolonged outflow of both may be involved with apoptosis. Elevated levels of these three elements in the CSF indicate a possible molecular pathway involved in the pathogenic process of the disease.

Zinc distribution in human prostate carcinoma cell line using synchrotron X‐ray microfluorescence

Cancer is a worldwide public health problem, and its incidence in the world grew by 20% in the last decade. Zinc, an essential trace element, is involved in many cellular processes. Concerning prostate cancer, Zn could inhibit the growth of tumor cells, by inducing cell cycle arrest or apoptosis. X‐ray microfluorescence (μXRF) is an elemental analysis technique that allows mapping biologically important elements at a submillimeter scale, with high sensitivity and negligible damage to the sample. In this study, we investigated cell viability through colorimetric cytotoxicity assay (MTT) and the behavior of human prostate cell lines obtained from normal (RWPE‐1) and tumorigenic (DU145) epithelium, in three‐dimensional spheroid cultures after supplementation with ZnCl2 for 24 and 48 hr using synchrotron μXRF. The measurements were performed at the Synchrotron Light National Laboratory (Campinas, Brazil). The results of μXRF showed that Zn intensity decreased in the DU145 tumor cells independent of supplementation or treatment time, and in normal cells, the intensity increased with supplementation and treatment time. The MTT assay results showed no significant differences, which indicates that the cell viability did not change. Our results indicate that μXRF can be used as an important tool to understand the mechanism of the loss of ability to capture zinc by prostate cancer cells. Copyright © 2017 John Wiley & Sons, Ltd.

Multielement concentration analysis of Swiss mice brains on experimental model of Alzheimer's disease induced by β‐amyloid oligomers

Alzheimer's disease (AD) is an insidious, progressive, and irreversible brain disorder that leads to memory loss and severe cognitive decline. However, the discovery that soluble β‐amyloid oligomers are potent central nervous system neurotoxins has led to a new view of AD pathogenesis. Metals, such as zinc, iron, and copper, are all increased in the aged brain, and the contents of those metals in the brain of AD patients are higher than non‐AD people. So, altered homeostasis of metal may result in the development of AD. Total reflection X‐ray fluorescence analysis (TXRF) is a multielement analytical technique, which can be used for elemental trace analysis. In this work, TXRF was used to evaluate the elemental concentration and distribution on mice brain regions. Three groups were studied: control, AD10, and AD100, being the two latter were given a single intracerebroventricular injection of 10 pmol and 100 pmol (β‐amyloid oligomers), respectively, to induce experimental AD. All samples were submitted to acid digestion. The TXRF measurements were performed at the X‐Ray Fluorescence Beamline at Brazilian National Synchrotron Light Laboratory, using a monochromatic beam (11.5 keV) for the excitation. It was possible to determine the concentrations of the following elements: P, S, K, Fe, Cu, and Zn. Results showed differences in the elemental concentration in some brain regions between the AD groups and the control group. Furthermore, the difference in the hypothalamus in AD10 groups, both female and male, suggests an association between AD and changes in these elements. Copyright © 2017 John Wiley & Sons, Ltd.

Multielement analysis in serum of healthy population of the metropolitan region of Rio de Janeiro in Brazil by SRTXRF

The determination of multielemental concentrations in physiological fluids is of considerable interest in clinical chemistry. Since it has been established these levels in human serum can be utilized as indicators for several pathological conditions. The multielement composition determination in human serum is important for the investigation of their role in human physiology and metabolism. From this research, more detailed studies on the health of individuals living in the metropolitan region of the state of Rio de Janeiro can be realized in order to improve the quality of life of these individuals. We studied 180 healthy volunteers (80 males and 100 females) aged 19 to 60 years. All the serum samples had been collected of people who live in the urban area of Rio de Janeiro City/Brazil. The measurements were performed at the X‐Ray Fluorescence Beamline at the Brazilian National Synchrotron Light Laboratory (LNLS), in Campinas, São Paulo. We were able to determine the concentrations of the following 11 elements: Na, P, S, Cl, K, Ca, Fe, Cu, Zn, Br and Rb. In this study, we used statistical analysis for the principal components. We were able to verify the presence of two factors that divided the elements into two groups: group 1 formed by elements: Cl, K, S and Ca and group 2 by elements Cu and Zn. Copyright © 2015 John Wiley & Sons, Ltd.

Study of the differential response to zinc of prostate cell spheroids using the synchrotron radiation X‐ray microfluorescence technique

X‐ray microfluorescence imaging technique has very often been used as a significant tool in biomedical research to obtain information regarding the distribution of elements in tissue and cell samples. This study investigated Zn distribution in benign prostate hyperplasia (BPH) and prostate cancer (DU145) cell spheroids and analyzed the different response to Zinc treatment using synchrotron X‐ray microfluorescence (μSRXRF). The μSRXRF measurements were performed at the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). The results showed non‐uniform distribution of Zn in all the spheroids analyzed. The differential response of DU145 and BPH cell spheroids to zinc suggests that there are different mechanisms of Zn accumulation in tumor cells. Copyright © 2015 John Wiley & Sons, Ltd.

Pair distribution function analysis at the Brazilian synchrotron light source

The XDS beamline of the Synchrotron Light National Laboratory (LNLS), was designed to take advantage of the 4T superconducting multipole wiggler inserted in the storage ring. This multipurpose beam line is employed for X-ray diffraction and X-ray absorption spectroscopy in the energy range between 5 and 30 keV. The X-ray diffraction patterns can be acquired in two different arrangements: a) Bragg-Brentano configuration, using a scintillation detector with an analyzer and b) Debye-Scherrer configuration, where the sample is mounted into capillaries and the diffraction pattern is acquired with an arrangement of 6 Mythen detectors or a scintillator. The sample can be measured at different atmospheres and temperatures. The viability of the beamline for pair distribution function analysis (PDF) was tested measuring two different standards: 1) Al2O3 and 2) BaTiO3. The patterns were acquired at room temperature using the two detection setups at an energy of 20 keV. The samples were mounted inside 0.3 mm boron-rich glass capillaries. In addition to the sample pattern, we also measured the empty capillary (background) to subtract it to the sample data. The acquired and normalized patterns were converted into total scattering PDF (G(r)) with the PDFgetX3.[1] The experimental G(r) was fitted with the PDFgui.[2] Both data sets were fitted in the corresponding structural phase with cell parameters close to the ones reported in the literature. In the special case of the BaTiO3 sample, it was very carefully modeled. We particularly focused in evaluating if we can discriminate the correct structural phase, as this sample presents different phases (orthorhombic, tetragonal and cubic). We could identify that the sample, at room temperature, was at the expected tetragonal phase. Finally, we will present a preliminary analysis of the following systems: Pb1-xRxZr1-yTiyO3 (R=La&Ba) and Ba1-xRxZr1-yTiyO3 (R=La&Ca) at the ferro- and paraelectric states by PDF.

Multielemental analysis of samples from patients with dermatological pathologies using synchrotron radiation

Using synchrotron radiation total X-ray fluorescence (SRTXRF) technique, the concentrations of trace elements were measured in four skin lesions: seborrheic keratosis, fibroepithelial polyp, cherry angioma and dermatosis papulosa nigra. The concentrations of P, S, K, Ca, Fe, Cu, Zn and Rb were evaluated in 62 pairs of lesions and healthy samples, each one having been collected from the same patient. The results revealed significant differences of P, Ca, K, Fe and Cu levels as well as a common trend in their variations between lesion and control samples among the skin diseases. This study revealed a powerful tool that can be useful for skin disorders research. The measurements were conducted at Brazilian National Synchrotron Light Laboratory (LNLS).

Determination of low Z in human serum of patients with Idiopathic Thrombocytopenic Purpura by total reflection X‐ray fluorescence

Idiopathic Thrombocytopenic Purpura (ITP) is a blood disorder characterized by an abnormal decrease in the number of platelets in the blood. In this work, 40 patients suffering from ITP and 60 healthy volunteers (Control Group) were analyzed. All the serum samples were collected from people who live in the urban area of Rio de Janeiro City/Brazil. The measurements were performed at the X‐ray fluorescence Beamline at the Brazilian National Synchrotron Light Laboratory, in Campinas, São Paulo using a monochromatic beam with maximum energy of 20 keV for the excitation and an Ultra‐LEGe detector with resolution of 148 eV at 5.9 keV. Standard solutions with Vanadium as internal standard were prepared for the calibration system. It was possible to determine the elemental concentrations of the following six elements: Na, P, S, Cl, K and Ca. The Student's t‐test was used to analyze significant differences (α = 0.05) between patients with ITP and Control Group. The elements that presented significant differences for the mean of their concentrations between each one from the ITP group and Control Group in µg g−1, were phosphorous (136 ± 12 and 92 ± 12), sulfur (1077 ± 97 and 847 ± 80), chlorine (2905 ± 385 and 2266 ± 378), potassium (137 ± 118 and 82 ± 15) and calcium (64 ± 7 and 44 ± 6), respectively. Copyright © 2013 John Wiley & Sons, Ltd.

Distribution of Fe, Cu and Zn in Cellular Spheroid Derived Human Prostate Tumor Cells by Synchrotron X-Ray Fluorescence

The formation of three-dimensional cell microspheres such as spheroids has attracted attention as a useful culture technique. In this study, we investigated the distribution (mapping) of Fe, Cu and Zn in cellular spheroid derived human prostate tumor cells. The measurements were performed in standard geometry of 45° incidence, exciting with a white beam collimated with the help of a 20 μm diameter optical capillary in the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). The results showed that all the elements analyzed presented non-uniform distribution. The spheroids analyzed showed different elemental distribution of Fe and Cu. Zinc concentrations were higher in the central regions of the spheroids analyzed.

The U11 PGM beam line at the Brazilian National Synchrotron Light Laboratory

We present the U11 beam line at the National Synchrotron Light Laboratory (LNLS), Brazil. This is the first undulator based beam line installed at the UVX storage ring. It is based on the collimated plane grating monochromator design, which allows for variation in included angle of the grating. The results obtained during the commissioning demonstrate a good overall performance in terms of energy resolution and beam size, in accord with the expected values obtained from ray tracing calculation.

Elemental concentration analysis in prostate tissues using total reflection X-ray fluorescence

Prostate cancer (PCa) currently represents the second most prevalent malignant neoplasia in men, representing 21% of all cancer cases. Benign Prostate Hyperplasia (BPH) is an illness prevailing in men above the age of 50, close to 90% after the age of 80. The prostate presents a high zinc concentration, about 10-fold higher than any other body tissue. In this work, samples of human prostate tissues with cancer, BPH and normal tissue were analyzed utilizing total reflection X-ray fluorescence spectroscopy using synchrotron radiation technique (SR-TXRF) to investigate the differences in the elemental concentrations in these tissues. SR-TXRF analyses were performed at the X-ray fluorescence beamline at Brazilian National Synchrotron Light Laboratory (LNLS), in Campinas, São Paulo. It was possible to determine the concentrations of the following elements: P, S, K, Ca, Fe, Cu, Zn and Rb. By using Mann–Whitney U test it was observed that almost all elements presented concentrations with significant differences (α=0.05) between the groups studied.

Determination of Cu/Zn and Fe in human serum of patients with sickle cell anemia using radiation synchrotron

In this work we analyzed serum samples from patients with Sickle Cell Anemia (SCA) using Total Reflection X-Ray Fluorescence using Synchrotron Radiation (SRTXRF). The SRTXRF measurements were performed at the X-Ray Fluorescence Beamline at the Brazilian National Synchrotron Light Laboratory (LNLS). We studied forty-three patients aged 18–50 suffering from SCA and sixty healthy volunteers aged 18–60. It was possible to determine the concentrations of the following elements: P, S, Cl, K, Ca, Fe, Cu, Zn, Br and Rb. Moreover, there are evidences of an association among Fe, Cu, Zn and Cu/Zn in the SCA pathogenesis process. The concentrations of Fe and Cu in the serum samples of patients with SCA were larger, 120% and 20%, respectively, when compared with the CG. The serum level Cu/Zn ratio was significantly higher (60%) in the serum samples from patients suffering from SCA than from the CG. Therefore, the Cu/Zn ratio can be used as an adjuvant index in enhancement for diagnosis of SCA.

Protein preparation, crystallization and preliminary X-ray analysis of Trypanosoma cruzi nucleoside diphosphate kinase 1.

The flagellated protozoan parasite Trypanosoma cruzi is the aetiological agent of Chagas disease. Nucleoside diphosphate kinases (NDPKs) are enzymes that are involved in energy management and nucleoside balance in the cell. T. cruzi TcNDPK1, a canonical isoform, was overexpressed in Escherichia coli as an N-terminally poly-His-tagged fusion protein and crystallized. Crystals grew after 72 h in 0.2 M MgCl(2), 20% PEG 3350. Data were collected to 3.5 A resolution using synchrotron X-ray radiation at the National Synchrotron Light Laboratory (Campinas, Brazil). The crystals belonged to the trigonal space group P3, with unit-cell parameters a = b = 127.84, c = 275.49 A. Structure determination is under way and will provide relevant information that may lead to the first step in rational drug design for the treatment of Chagas disease.

High resolution study of Kβ′ and Kβ1,3 X-ray emission lines from Mn-compounds

Abstract High-resolution Kβ emission spectra of several manganese compounds were measured in order to characterize the dependence of the Kβ′ and Kβ 1,3 features, on the chemical environment. High resolution spectra were obtained using a non-conventional spectrometer based on quasi-back-diffraction geometry at National Synchrotron Light Laboratory (LNLS). It was found that the energy of the Kβ′ satellite structure relative to the main Kβ 1,3 line decreases linearly with the formal oxidation state for Mn–O systems. A noticeable dispersion of the relative Kβ′ energy for different Mn 2+ compounds could be observed. The dependence of the Kβ′ satellite line on the net charge and the effective 3 d spin in Mn 2+ compounds was investigated. Calculations of the net charge and the effective 3 d spin were performed within the density-functional theory using the package SIESTA. A direct relation between this dispersion and the effective Mn 3 d spin was found.

X-ray fluorescence with synchrotron radiation to elemental analysis of lead and calcium content of primary teeth

Primary teeth were analyzed by micro-SRXRF. The aim of this study was to determine the elemental distribution of lead and calcium in different regions of primary incisor of children living in a notoriously contaminated area (Santo Amaro da Purificação, Bahia State, Brazil). The measurements were performed in standard geometry of 45° incidence, exciting with a white beam and using a conventional system collimation (orthogonal slits) in the XRF beamline at the Synchrotron Light National Laboratory (Campinas, Brazil).

Latest developments and opportunities for 3D analysis of biological samples by confocal μ-XRF

X-ray fluorescence analysis performed with a primary radiation focused in the micrometer range is known as micro-X-ray fluorescence (μ-XRF). It is characterized by a penetration depth higher than other micro-analytical methods, reaching hundreds of micrometers in biological samples. This characteristic of the X-ray beam can be employed in 3D analysis. An innovative method to perform 3D analysis by μ-XRF is the so-called confocal setup. The confocal setup consists of X-ray lenses in the excitation as well as in the detection channel. In this configuration, a micro-volume defined by the overlap of the foci of both X-ray lenses is analyzed. Scanning this micro-volume through the sample can be used to perform a study in three dimensions. At present, X-ray lenses used in confocal μ-XRF experiments are mainly glass capillaries and polycapillaries. Glass capillaries are used in the excitation channel with sources of high photon flux like synchrotron radiation. Half polycapillaries or conical polycapillary concentrators are used almost exclusively in the detection channel. Spatial resolution of the confocal μ-XRF depends on the dimensions of the foci of both X-ray lenses. The overlap of these foci forms an ellipsoid which is the probing volume of the confocal setup. The axis length of the probing volume reported in confocal μ-XRF experiments are of order of few tens of micrometer. In our confocal setup, we used a commercial glass monocapillary in the excitation channel and a monolithic half polycapillary in the detection channel. The polycapillary was home-made by means of drawing of multibundles of glass capillaries in a heating furnace. The experiment was carried out at the beamline D09B-XRF of the Synchrotron Light National Laboratory (Laboratório Nacional de Luz Síncrotron, LNLS) using white beam. A model for the theoretical description of X-ray fluorescence intensity registered by confocal μ-XRF was introduced by Malzer and Kanngieβer [2005. A model for the confocal volume of 3D micro-X-ray fluorescence spectrometer. Spectrochim. Acta B 60, 1334–1341]. These authors showed that the measured scanning of fluorescent intensity is the spatial convolution of the sensitivity and the X-ray fluorescence emission rate detected. In a previous work we demonstrated that the convolution theorem can simplify the calibration and quantification process in confocal μ-XRF. In the present work, we applied these ideas to analyze samples of aquatic plants and a sample of tooth by confocal μ-XRF. We showed that confocal μ-XRF can be successfully applied to help the study of the effects of water bioremediation on aquatic plants. We also showed that confocal μ-XRF can be used to make topological studies of teeth.

Topographic Trace-Elemental Analysis in the Brain of Wistar Rats by X-ray Microfluorescence with Synchrotron Radiation

Knowledge about the spatial distribution and the local concentration of trace elements in tissues is of great importance, since trace elements are involved in many biological functions of living organisms. However, there are few methods available to measure the spatial (two (three)-dimensional) elemental distribution in animal brain. X-ray microfluorescence with synchrotron radiation is a multielemental mapping technique, which was used in this work to determine the topographic of iron, zinc and copper in coronal sections of female Wistar rats of different ages. Young (14 days old) and middle-aged (20 months old) rats (n = 8) were analyzed. The measurements were carried out at the XRF beam line at the Synchrotron Light National Laboratory (Campinas, Brazil). Two-dimensional scanning was performed in order to study the tendency of elemental concentration variation. The acquisition time for each pixel was 10 s/step and the step size was 300 microm/step in both directions. It was observed that the iron distribution was more conspicuous in the cortical area, thalamus and bellow the thalamus. On the other hand, the zinc distribution was more pronounced in the hippocampus. The iron, copper and zinc levels increased with advancing age. Therefore, this study reinforces the idea that these elements are involved in the chemical mechanisms of the brain that induce some neurological diseases, since they are also present in high levels in specific areas of the brain, such as the hippocampus and the substantia nigra of patients with these disorders.