RESUMO
INTRODUCTION: The aim of this in vitro study was to assess the suitability of high-resolution time-of-flight secondary ion mass spectrometry (ToF-SIMS) for visualizing cross-sectional changes in human enamel microstructure and chemical composition during treatment and remineralization cycling of artificially generated caries lesions underneath an artificial plaque. METHODS: Treatments consisted of exposure to twice daily toothpaste/water slurries prepared from 0, 1,100, and 5,000 µg/g fluoride (F) NaF/silica toothpastes. In addition, treatments with slurries prepared from 1,100 µg/g F SnF2/silica toothpastes were done using 44Ca in the remineralization solution to allow for differentiation of newly formed mineral and exploration of incorporated metal dopants using ToF-SIMS. Complementary microhardness, scanning electron microscopy, and high-resolution transmission electron microscopy (HR-TEM) investigations were performed on enamel cross sections. RESULTS: HR-TEM was used for the first time to determine the change in crystallinity during remineralization revealing distinct microstructural zones within one lesion. Chemical mapping using ToF-SIMS demonstrated that the distribution of F, while observed primarily in the new mineral phase, was widespread throughout the lesion with 44Ca substantially limited to the remineralizing mineral. Both penetrated the inter-rod spaces of the sound enamel illustrating how acid damage propagates into the native mineral as the caries lesion deepens. HR-TEM examination revealed different regions within the lesion characterized by distinct micro- and ultrastructures. Importantly, HR-TEM revealed a return of crystallinity following remineralization. F dose-response observations verified the ability of these high-resolution techniques to differentiate remineralization efficacy. CONCLUSION: The collective results provided new insights such as the visualization of F or calcium penetration pathways, as well as new tools to study the caries process.
Assuntos
Esmalte Dentário , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Espectrometria de Massa de Íon Secundário , Remineralização Dentária , Remineralização Dentária/métodos , Esmalte Dentário/ultraestrutura , Esmalte Dentário/química , Espectrometria de Massa de Íon Secundário/métodos , Humanos , Cremes Dentais/uso terapêutico , Cremes Dentais/química , Fluoreto de Sódio/uso terapêutico , Dureza , Cálcio/análise , Cariostáticos/uso terapêutico , Cárie Dentária/diagnóstico por imagem , Cárie Dentária/patologia , Fluoretos de Estanho/uso terapêutico , Fluoretos , Dióxido de Silício , CristalizaçãoRESUMO
Identifying characteristic extracellular matrix (ECM) variants is a key challenge in mechanistic biology, bioengineering, and medical diagnostics. The reported study demonstrates the potential of time-of-flight secondary ion mass spectrometry (ToF-SIMS) to detect subtle differences between human mesenchymal stromal cell (MSC)-secreted ECM types as induced by exogenous stimulation or emerging pathology. ToF-SIMS spectra of decellularized ECM samples are evaluated by discriminant principal component analysis (DPCA), an advanced multivariate analysis technique, to decipher characteristic compositional features. To establish the approach, signatures of major ECM proteins are determined from samples of pre-defined mixtures. Based on that, sets of ECM variants produced by MSCs in vitro are analyzed. Differences in the content of collagen, fibronectin, and laminin in the ECM resulting from the combined supplementation of MSC cultures with polymers that induce macromolecular crowding and with ascorbic acid are detected from the DPCA of ToF-SIMS spectra. The results are verified by immunostaining. Finally, the comparative ToF-SIMS analysis of ECM produced by MSCs of healthy donors and patients suffering from myelodysplastic syndrome display the potential of the novel methodology to reveal disease-associated alterations of the ECM composition.
Assuntos
Células-Tronco Mesenquimais , Espectrometria de Massa de Íon Secundário , Humanos , Espectrometria de Massa de Íon Secundário/métodos , Análise de Componente Principal , Análise Multivariada , Matriz ExtracelularRESUMO
The mass and volume concentration of nanoplastics is extremely low, but incredibly high in terms of surface area; this is expected to increase their toxicity through the ab/adsorption and transport of chemical co-pollutants such as trace metals. In this context, we studied the interactions between nanoplastics model materials functionalized with carboxylated groups, with either smooth or raspberry-like surface morphologies, and copper as representative of trace metals. For this purpose, a new methodology, using two complementary surface analysis techniques: Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) and X-ray Photoelectron Spectroscopy (XPS) was developed. In addition, inductively coupled plasma mass spectrometry (ICP-MS) was used to quantify the total mass of sorbed metal on the nanoplastics. This innovative analytical approach from the top surface to the core of nanoplastics demonstrated not only the interactions with copper at the surface level, but also the ability of nanoplastics to absorb metal at their core. Indeed, after 24 h of exposition, the copper concentration at the nanoplastic surface remained constant due to saturation whereas the copper concentration inside the nanoplastic keeps increasing with the time. The sorption kinetic was evaluated to increase with the density of charge of the nanoplastic and the pH. This study confirmed the ability of nanoplastics to act as metal pollutant carriers by both adsorption and absorption phenomena.
Assuntos
Microplásticos , Oligoelementos , Cobre/química , Análise Espectral , Espectrometria de Massa de Íon Secundário/métodos , AdsorçãoRESUMO
Traces of lipids, absorbed and preserved for millennia within the inorganic matrix of ceramic vessels, act as molecular fossils and provide manifold information about past people's subsistence, diet, and rituals. It is widely assumed that lipids become preserved after adsorption into nano- to micrometer-sized pores, but to this day the distribution of these lipids in the ceramics was virtually unknown, which severely limits our understanding about the process of lipid preservation. Here we use secondary ion mass spectrometry (SIMS) imaging for direct in situ analysis of lipids absorbed in 700- to 2,000-y-old archaeological pottery. After sectioning from larger sherds, wall cross-sections of smaller fragments were used for SIMS analysis. Lipids were found in relatively large zones of 5- to 400-µm diameter, which does not support the notion of absorption only into individual nanometer-scale pores but indicates that more macroscopic structures in the ceramics are involved in lipid preservation as well. Furthermore, lipids were found concentrated on calcium carbonate inclusions in the ceramics, which suggests that precipitation of fatty acids as calcium salts is an important aspect of lipid preservation in archaeological samples. This has important implications for analytical methods based on extraction of lipids from archaeological ceramics and needs to be considered to maximize the yield and available information from each unique sample.
Assuntos
Arqueologia/métodos , Cerâmica/química , Argila/química , Lipídeos , Espectrometria de Massa de Íon Secundário/métodos , Cerâmica/história , Culinária/história , História Antiga , Humanos , Lipídeos/análise , Lipídeos/química , Imagem Molecular , Reino UnidoRESUMO
This study is aimed to observe changes in fatty acid profiles by time of flight secondary ion mass spectrometry (ToF-SIMS) in breast muscle tissues of broilers. Four different groups were identified. The source of fat in group I was soy oil (rich in linoleic acid, ω-6), group II received linseed oil (ω-3), and the third group was fed a mixture of the two mentioned oils. Broilers in the control group were fed with beef tallow, used in mass commercial production. The results reveal that the use of vegetable oils in animal nutrition determines the lipid profile of fatty acids. ToF-SIMS measurements showed that the lipid profile of muscle fibers and intramuscular fat reflect the composition of fats used as feed additives. In both structures, the ratio of ω-6/ω-3 fatty acids, which is most favorable for human health, was found in the groups in which a mixture of vegetable oils and a supplement of linseed oil were used.
Assuntos
Ácidos Graxos/análise , Carne/análise , Músculo Esquelético/química , Espectrometria de Massa de Íon Secundário/métodos , Ração Animal , Animais , Galinhas , Gorduras na Dieta , Óleos de PlantasRESUMO
Single-cell measurements of biochemical processes have advanced our understanding of cellular physiology in individual microbes and microbial populations. Due to methodological limitations, little is known about single-cell phosphorus (P) uptake and its importance for microbial growth within mixed field populations. Here, we developed a nanometer-scale secondary ion mass spectrometry (nanoSIMS)-based approach to quantify single-cell P uptake in combination with cellular CO2 and N2 fixation. Applying this approach during a harmful algal bloom (HAB), we found that the toxin-producer Nodularia almost exclusively used phosphate for growth at very low phosphate concentrations in the Baltic Sea. In contrast, the non-toxic Aphanizomenon acquired only 15% of its cellular P-demand from phosphate and ~85% from organic P. When phosphate concentrations were raised, Nodularia thrived indicating that this toxin-producer directly benefits from phosphate inputs. The phosphate availability in the Baltic Sea is projected to rise and therefore might foster more frequent and intense Nodularia blooms with a concomitant rise in the overall toxicity of HABs in the Baltic Sea. With a projected increase in HABs worldwide, the capability to use organic P may be a critical factor that not only determines the microbial community structure, but the overall harmfulness and associated costs of algal blooms.
Assuntos
Aphanizomenon/crescimento & desenvolvimento , Aphanizomenon/metabolismo , Metabolismo , Nodularia/crescimento & desenvolvimento , Nodularia/metabolismo , Fósforo/metabolismo , Água do Mar/microbiologia , Análise de Célula Única/métodos , Espectrometria de Massa de Íon Secundário/métodosRESUMO
The largest lake transgression event (LTE) associated with lake anoxic events (LAE) and periodic seawater incursion events (SWIE) in the Songliao Basin, northeastern China, occurred during deposition of the Cretaceous Nenjiang Formation. The Yaojia-Nenjiang Formation boundary (YNB) marks the beginning of the LTE, as well as LAE and SWIE. However, there is an absence of direct radioisotopic dating, and therefore the age of the YNB, as well as the beginning of LTE, together with their relationship with other geological events, is strongly debated. Here we present a new SIMS U-Pb zircon age from the lowermost Nenjiang Formation. The bentonite bed located 9.88 m above the YNB of the X1-4 borehole was analyzed. Twenty-five analyses of 25 zircons were conducted, which produced a weighted mean age of 85.5±0.6 Ma (MSWD = 0.87). Based on the average sediment accumulation rate, the age of the YNB is suggested to be 85.7 Ma, indicating that the LTE began in the Early Santonian. The new ages provide a precise chronostratigraphic framework for climatic and geological events. Our new results imply that the beginning of the LTE, LAE and SWIE occurred almost simultaneously with short-term sea level rise, and probably had a close relationship with OAE3.
Assuntos
Lagos/química , Chumbo/análise , Espectrometria de Massa de Íon Secundário/métodos , Urânio/análise , China , Geografia , Luminescência , Fatores de TempoRESUMO
Time of flight secondary ion mass spectrometry (ToF-SIMS) was used to image the lipid localization in brain tissue sections from rats fed specially processed cereals (SPC). An IonTof 5 instrument equipped with a Bi cluster ion gun was used to analyze the tissue sections. Data from 15 brain samples from control and cereal-fed rats were recorded and exported to principal components analysis (PCA). The data clearly show changes of certain lipids in the brain following cereal feeding. PCA score plots show a good separation in lipid distribution between the control and the SPC-fed group. The loadings plot reveal that the groups separated mainly due to changes in cholesterol, vitamin E and c18:2, c16:0 fatty acid distribution as well as some short chain monocarboxylic fatty acid compositions. These insights relate to the working mechanism of SPC as a dietary supplement. SPC is thought to activate antisecretory factor (AF), an endogenous protein with regulatory function for inflammation and fluid secretion. These data provide insights into lipid content in brain following SPC feeding and suggest a relation to activating AF.
Assuntos
Lipídeos/química , Neurônios/metabolismo , Animais , Colesterol/metabolismo , Grão Comestível/metabolismo , Ácidos Graxos/metabolismo , Alimentos , Inflamação/metabolismo , Masculino , Neuropeptídeos/metabolismo , Análise de Componente Principal/métodos , Ratos , Ratos Sprague-Dawley , Espectrometria de Massa de Íon Secundário/métodos , Vitamina E/metabolismoRESUMO
In order to utilize complementary imaging techniques to supply higher resolution data for fusion with secondary ion mass spectrometry (SIMS) chemical images, there are a number of aspects that, if not given proper consideration, could produce results which are easy to misinterpret. One of the most critical aspects is that the two input images must be of the same exact analysis area. With the desire to explore new higher resolution data sources that exists outside of the mass spectrometer, this requirement becomes even more important. To ensure that two input images are of the same region, an implementation of the insight segmentation and registration toolkit (ITK) was developed to act as a preprocessing step before performing image fusion. This implementation of ITK allows for several degrees of movement between two input images to be accounted for, including translation, rotation, and scale transforms. First, the implementation was confirmed to accurately register two multimodal images by supplying a known transform. Once validated, two model systems, a copper mesh grid and a group of RAW 264.7 cells, were used to demonstrate the use of the ITK implementation to register a SIMS image with a microscopy image for the purpose of performing image fusion.
Assuntos
Processamento de Imagem Assistida por Computador/métodos , Imagem Multimodal/métodos , Espectrometria de Massa de Íon Secundário/métodos , Animais , Linhagem Celular , Cobre , Macrófagos , CamundongosRESUMO
Dietary intake of omega-3 fatty acids is associated with considerable health benefits, including the prevention of metabolic disorders such as cardiovascular disease and type 2 diabetes. Furthermore, incorporation of the main omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), at the systemic level has been found to be more efficient when these fatty acids are supplied in the form of marine phospholipids compared to triglycerides. In this work, the uptake of omega-3 fatty acids and their incorporation in specific lipids were studied in adipose, skeletal muscle, and liver tissues of mice given high-fat diets with or without omega-3 supplements in the form of phospholipids or triglycerides using time-of-flight secondary ion mass spectrometry (TOF-SIMS). The results demonstrate significant uptake of EPA and DHA, and the incorporation of these fatty acids in specific lipid molecules, in all three tissue types in response to the dietary omega-3 supplements. Moreover, the results indicate reduced concentrations of arachidonic acid (AA) and depletion of lipids containing AA in tissue samples from mice given supplementary omega-3, as compared to the control mice. The effect on the lipid composition, in particular the DHA uptake and AA depletion, was found to be significantly stronger when the omega-3 supplement was supplied in the form of phospholipids, as compared to triglycerides. TOF-SIMS was found to be a useful technique for screening the lipid composition and simultaneously obtaining the spatial distributions of various lipid classes on tissue surfaces.
Assuntos
Ácidos Graxos Ômega-3/análise , Ácidos Graxos Ômega-3/metabolismo , Espectrometria de Massa de Íon Secundário/métodos , Tecido Adiposo/química , Tecido Adiposo/metabolismo , Animais , Dieta Hiperlipídica , Fígado/química , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/química , Músculo Esquelético/metabolismoRESUMO
Time-of-flight secondary ion mass spectrometry (ToF-SIMS) was used to characterize the freeze-fracturing process of human epithelial PANC-1 and UROtsa cells. For this purpose, phosphatidylcholine, sphingomyelin, phosphatidylethanolamine, and phosphatidylserine standard samples were investigated to find specific signals with both high specificity and signal intensity. The results were used to investigate single cells of subconfluent cell layers prepared with a special silicon wafer sandwich preparation technique. This freeze-fracturing technique strips cell membranes off the cells, isolating them on opposing silicon wafer substrates. Criteria were found for defining regions with stripped off cell membranes and, on the opposing wafer, complementary regions with the remaining cells. Measured ethanolamine/choline and serine/choline ratios in these regions clearly showed that in the freeze-fracturing process, the lipid bilayer of the plasma membrane is split along its central zone. Accordingly, only the outer lipid monolayer is stripped off the cell, while the inner lipid monolayer remains attached to the cell on the opposing wafer, thus allowing detailed analysis of a single lipid monolayer. Furthermore, it could be shown that using different washing procedures did not influence the transmembrane lipid distribution. Under optimized preparation conditions, it became feasible to detect lipids with a lateral resolution of approximately 100 nm. The data indicate that ToF-SIMS would be a very useful technique to study with very high lateral resolution changes in lipid composition caused, for example, by lipid storage diseases or pharmaceuticals that interfere with the lipid metabolism.
Assuntos
Membrana Celular/química , Células Epiteliais/química , Espectrometria de Massa de Íon Secundário/métodos , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Células Epiteliais/metabolismo , Técnica de Fratura por Congelamento , Humanos , Bicamadas Lipídicas/química , Metabolismo dos Lipídeos , Lipídeos/químicaRESUMO
Stable isotopes are ideal labels for studying biological processes because they have little or no effect on the biochemical properties of target molecules. The NanoSIMS is a tool that can image the distribution of stable isotope labels with up to 50 nm spatial resolution and with good quantitation. This combination of features has enabled several groups to undertake significant experiments on biological problems in the last decade. Combining the NanoSIMS with other imaging techniques also enables us to obtain not only chemical information but also the structural information needed to understand biological processes. This article describes the methodologies that we have developed to correlate atomic force microscopy and backscattered electron imaging with NanoSIMS experiments to illustrate the imaging of stable isotopes at molecular, cellular, and tissue scales. Our studies make it possible to address 3 biological problems: (1) the interaction of antimicrobial peptides with membranes; (2) glutamine metabolism in cancer cells; and (3) lipoprotein interactions in different tissues.
Assuntos
Glutamina/metabolismo , Microscopia de Força Atômica/métodos , Neoplasias/metabolismo , Espectrometria de Massa de Íon Secundário/métodos , Peptídeos Catiônicos Antimicrobianos/metabolismo , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Humanos , Marcação por Isótopo/métodos , Lipoproteínas/metabolismo , Nanotecnologia/métodos , Neoplasias/patologia , Distribuição TecidualRESUMO
Secondary ion mass spectrometry (SIMS) was used in combination with alpha track detection for the efficient analysis of uranium-bearing particles with higher (235)U abundances in environmental samples. A polycarbonate film containing particles was prepared and placed in contact with a CR-39 plastic detector. After exposure for 28 days, the detector was etched in a NaOH solution and each uranium-bearing particle was identified through observation of the alpha tracks recorded in the detector. A portion of the film containing each uranium-bearing particle was cut out and put onto a glassy carbon planchet. The films on the planchet were decomposed through plasma ashing for subsequent uranium abundance ratio analysis with SIMS. The alpha track-SIMS analysis of 10 uranium-bearing particles in a sample taken from a nuclear facility enabled n((235)U)/n((238)U) abundance ratios in the range 0.0072-0.25 to be detected, which were significantly higher than those obtained by SIMS without alpha track detection. The duration of the whole analytical process for analysis of 10 particles was about 32 days. The detection efficiency was calculated to be 27.1±6.5%, based on the analysis of the particles in uranium reference materials. The detection limits, defined as the diameter of the particle which produces alpha tracks more than one for a 28-days exposure, were estimated to be 0.8, 0.9, 1.1, 2.1 and 3.0 µm for the particles having the same uranium abundance ratios with NBL CRM U850, U500, U350, U050 and U010 reference materials, respectively. The use of alpha track detection for subsequent SIMS analysis is an inexpensive and an efficient way to measure uranium-bearing particles with higher (235)U abundances.
Assuntos
Poluentes Ambientais/análise , Espectrometria de Massa de Íon Secundário/métodos , Urânio/análise , Limite de Detecção , Polietilenoglicóis/químicaRESUMO
Time-of-flight secondary ion mass spectrometry (ToF-SIMS) is a well-established technique in material sciences but has not yet been widely explored for implementation in life sciences. Here, we demonstrate the applicability and advantages of ToF-SIMS analysis for the study of minerals and biomolecules in osseous tissue. The locally resolved analysis of fragment ions deriving from the sample surface enables imaging and differentiation of bone tissue and facilitates histology on non-stained cross sections. In a rat model, bilateral ovariectomy combined with either a multi-deficiency diet or steroid treatment was carried out to create osteoporotic conditions. We focused our study on the Ca content of the mineralized tissue and monitored its decline. Calcium mass images of cross sections show the progressive degenerative changes in the bone. We observed a decreased Ca concentration in the edge region of the trabeculae and a decline in the Ca/P ratio. Additionally, we focused on the non-mineralized matrix and identified fragment ions that are characteristic for the collagen matrix. We observed trabeculae with wide ranges of non-mineralized collagen for the diet group owing to an impaired mineralization process. Here, the advantage of coeval monitoring of collagen and minerals indicated an osteomalacic model rather than an osteoporotic one.
Assuntos
Densidade Óssea , Cálcio/metabolismo , Colágeno/metabolismo , Vértebras Lombares/metabolismo , Osteoporose Pós-Menopausa/metabolismo , Espectrometria de Massa de Íon Secundário/métodos , Animais , Modelos Animais de Doenças , Feminino , Humanos , Vértebras Lombares/patologia , Osteoporose Pós-Menopausa/patologia , Ovariectomia , Fósforo/metabolismo , Ratos , Fatores de TempoRESUMO
UNLABELLED: The interrelation of calcium and phosphorus was evaluated as a function of bone material quality in femoral heads from male fragility fracture patients via surface analytical imaging as well as scanning microscopy techniques. A link between fragility fractures and increased calcium to phosphorus ratio was observed despite normal mineralization density distribution. INTRODUCTION: Bone fragility in men has been recently recognized as a public health issue, but little attention has been devoted to bone material quality and the possible efficacy in fracture risk prevention. Clinical routine fracture risk estimations do not consider the quality of the mineralized matrix and the critical role played by the different chemical components that are present. This study uses a combination of different imaging and analytical techniques to gain insights into both the spatial distribution and the relationship of phosphorus and calcium in bone. METHODS: X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry imaging techniques were used to investigate the relationship between calcium and phosphorus in un-embedded human femoral head specimens from fragility fracture patients and non-fracture age-matched controls. The inclusion of the bone mineral density distribution via backscattered scanning electron microscopy provides information about the mineralization status between the groups. RESULTS: A link between fragility fracture and increased calcium and decreased phosphorus in the femoral head was observed despite normal mineralization density distribution. Results exhibited significantly increased calcium to phosphorus ratio in the fragility fracture group, whereas the non-fracture control group ratio was in agreement with the literature value of 1.66 M ratio in mature bone. CONCLUSIONS: Our results highlight the potential importance of the relationship between calcium and phosphorus, especially in areas of new bone formation, when estimating fracture risk of the femoral head. The determination of calcium and phosphorus fractions in bone mineral density measurements may hold the key to better fracture risk assessment as well as more targeted therapies.
Assuntos
Cálcio/análise , Fraturas do Colo Femoral/metabolismo , Cabeça do Fêmur/química , Fraturas por Osteoporose/metabolismo , Fósforo/análise , Idoso , Idoso de 80 Anos ou mais , Artroplastia de Quadril , Estudos de Casos e Controles , Fraturas do Colo Femoral/patologia , Fraturas do Colo Femoral/cirurgia , Cabeça do Fêmur/ultraestrutura , Humanos , Masculino , Microscopia Eletrônica de Varredura/métodos , Fraturas por Osteoporose/patologia , Fraturas por Osteoporose/cirurgia , Espectroscopia Fotoeletrônica/métodos , Espectrometria de Massa de Íon Secundário/métodosRESUMO
A multiresidue analytical method has been developed to simultaneously determine the residues of 20 organochlorine pesticides in Herba epimedii, a traditional Chinese medicine. The 20 pesticides are included in the list of regulated substances by the Food and Environmental Hygiene Department in Hong Kong. The method consists of solid-phase extraction for cleanup of samples and GC coupled on-line with negative chemical ion-MS to analyze the target pesticides. The chromatographic separation of the compounds was carried out on a DB-1701 column by using an optimal temperature program, and the quantitative analysis was conducted by selected ion monitoring. The LOD and LOQ values fell in the range of 0.0555-5.8821 and 0.1241-17.9333 ng/g, respectively. The average recoveries were between 75.4 and 90.7% (n=5) for the 20 organochlorine pesticides. The developed method proved to be reliable and accurate, and permits rapid determination of the 20 organochlorine pesticides in one run.
Assuntos
Epimedium/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Hidrocarbonetos Clorados/análise , Praguicidas/análise , Espectrometria de Massa de Íon Secundário/métodos , Contaminação de Medicamentos , Medicamentos de Ervas Chinesas/análise , Humanos , Hidrocarbonetos Clorados/química , Estrutura Molecular , Praguicidas/química , Plantas Medicinais/química , Extração em Fase SólidaRESUMO
*Cereals are an important source of selenium (Se) to humans and many people have inadequate intakes of this essential trace element. Conversely, arsenic (As) is toxic and may accumulate in rice grain at levels that pose a health risk. Knowledge of the localization of selenium and arsenic within the cereal grain will aid understanding of their deposition patterns and the impact of processes such as milling. *High-resolution secondary ion mass spectrometry (NanoSIMS) was used to determine the localization of Se in wheat (Triticum aestivum) and As in rice (Oryza sativa). Combined synchrotron X-ray fluorescence (S-XRF) and NanoSIMS analysis utilized the strengths of both techniques. *Selenium was concentrated in the protein surrounding the starch granules in the starchy endosperm cells and more homogeneously distributed in the aleurone cells but with Se-rich hotspots. Arsenic was concentrated in the subaleurone endosperm cells in association with the protein matrix rather than in the aleurone cells. NanoSIMS indicated that the high intensity of As identified in the S-XRF image was localized in micron-sized hotspots near the ovular vascular trace and nucellar projection. *This is the first study showing subcellular localization in grain samples containing parts per million concentrations of Se and As. There is good quantitative agreement between NanoSIMS and S-XRF.
Assuntos
Arsênio/análise , Oryza/química , Sementes/química , Selênio/análise , Triticum/química , Endosperma/química , Endosperma/citologia , Oryza/citologia , Proteínas/química , Sementes/citologia , Espectrometria de Massa de Íon Secundário/métodos , Espectrometria por Raios X/métodos , Triticum/citologiaRESUMO
Lactobacillus sakei is a lactic acid bacterium naturally found on meat. Although it is generally acknowledged that lactic acid bacteria are rare species in the microbial world which do not have iron requirements, the genome sequence of L. sakei 23K has revealed quite complete genetic equipment dedicated to transport and use of this metal. Here, we aimed to investigate which iron sources could be used by this species as well as their role in the bacterium's physiology. Therefore, we developed a microscopy approach based on electron energy loss spectroscopy (EELS) analysis and nano-scale secondary-ion mass spectrometry (SIMS) in order to analyze the iron content of L. sakei cells. This revealed that L. sakei can use iron sources found in its natural ecosystem, myoglobin, hemoglobin, hematin, and transferrin, to ensure long-term survival during stationary phase. This study reveals that analytical image methods (EELS and SIMS) are powerful complementary tools for investigation of metal utilization by bacteria.
Assuntos
Ferro/metabolismo , Lactobacillus/metabolismo , Carne/microbiologia , Espectrometria de Massa de Íon Secundário/métodos , Espectroscopia de Perda de Energia de Elétrons/métodos , Catalase/metabolismo , Heme/metabolismo , Transferrina/metabolismoRESUMO
Lysaketotriol (1) and lysaketodiol (2), along with two previously known polyhydroxylated steroidal sulfates (3, 4), were isolated from the ethanolic extract of ambulakrums separated from the arms of the starfish Lysastrosoma anthosticta (Sea of Japan). The chemical structures have been elucidated using 1D and 2D NMR spectroscopy (1H, 13C, DEPT, COSY-45, HSQC and HMBC) and mass spectrometry. Compound 1 produced moderate stimulation of lysosomal activity in mouse splenocytes and compounds 1-2 showed immunomodulatory activity.
Assuntos
Estrelas-do-Mar/química , Esteroides/isolamento & purificação , Sulfatos/isolamento & purificação , Animais , Linfócitos/efeitos dos fármacos , Linfócitos/fisiologia , Macrófagos/efeitos dos fármacos , Macrófagos/fisiologia , Espectroscopia de Ressonância Magnética/métodos , Camundongos , Modelos Moleculares , Estrutura Molecular , Oceano Pacífico , Saponinas/química , Saponinas/isolamento & purificação , Saponinas/farmacologia , Espectrometria de Massa de Íon Secundário/métodos , Baço , Esteroides/química , Esteroides/farmacologia , Sulfatos/química , Sulfatos/farmacologiaRESUMO
This paper reports a method for the analysis of secondary metabolites stored in glandular trichomes, employing negative ion 'chip-based' nanospray tandem mass spectrometry. The analyses of glandular trichomes from Lychnophora ericoides, a plant endemic to the Brazilian 'cerrado' and used in traditional medicine as an anti-inflammatory and analgesic agent, led to the identification of five flavonoids (chrysin, pinocembrin, pinostrobin, pinobanksin and 3-O-acetylpinobanksin) by direct infusion of the extracts of glandular trichomes into the nanospray ionisation source. All the flavonoids have no oxidation at ring B, which resulted in a modification of the fragmentation pathways compared with that of the oxidised 3,4-dihydroflavonoids already described in the literature. The absence of the anti-inflammatory and antioxidant di-C-glucosylflavone vicenin-2, or any other flavonoid glycosides, in the glandular trichomes was also demonstrated. The use of the 'chip-based' nanospray QqTOF apparatus is a new fast and useful tool for the identification of secondary metabolites stored in the glandular trichomes, which can be useful for chemotaxonomic studies based on metabolites from glandular trichomes.