Person: Gómez Gómez, Beatriz
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Beatriz
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Gómez Gómez
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Universidad Complutense de Madrid
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Item Selenium and tellurium-based nanoparticles as interfering factors in quorum sensing-regulated processes: violacein production and bacterial biofilm formation(Metallomics, 2019) Gómez Gómez, Beatriz; Arregui García-Roves, Lucía; Serrano Barrero, Susana Lourdes; Santos de la Sen, Antonio; Pérez Corona, María Teresa; Madrid Albarrán, María YolandaA cell-to-cell communication system called quorum sensing (QS) promotes the transcription of certain target genes in bacterial cells leading to the activation of different cellular processes, some of them related to bacterial biofilm formation. The formation of bacterial biofilms favours antibiotic resistance, which is nowadays a significant public-health problem. In this study, the effect of selenium (SeNPs) and tellurium (TeNPs) nanoparticles was examined in two bacterial processes mediated by QS: violacein production by Chromobacterium violaceum and biofilm formation by Pseudomonas aeruginosa. For this purpose, quantification of the pigment production in the presence of these nanoparticles was monitored using the C. violaceum strain. Additionally, a combination of different microscopical imaging techniques was applied to examine the changes in the 3D biofilm structure of P. aeruginosa, which were quantified through performing architectural metric calculations (substratum area, cell area coverage and biovolume). SeNPs produce an 80% inhibition in the violacein production by C. violaceum and a significant effect on the P. aeruginosa biofilm architecture (a reduction of 80% in the biovolume of the bacterial biofilm was obtained). TeNPs similarly affect violacein production and the P. aeruginosa biofilm structure but at lower concentration levels. The results obtained suggest an important disruption of the QS signalling system by SeNPs and TeNPs, supporting nanotechnology as a promising tool to fight against the emerging problem of bacterial resistance related to bacterial biofilm formation.Item Fate and effect of in-house synthesized tellurium based nanoparticles on bacterial biofilm biomass and architecture. Challenges for nanoparticles characterization in living systems(Science of the Total Environment, 2020) Gómez Gómez, Beatriz; Sanz Landaluce, Jon; Pérez Corona, María Teresa; Madrid Albarrán, María YolandaThe unexpected impact of nanoparticles on environment and human health remains as a matter of concern. In this sense, understanding the interaction between nanoparticles and biological indicators such as microorganism may help to understand their fate and effect in environmental systems. However, the adverse effect of nanoparticles greatly depends on their properties and, therefore, a precise evaluation of nanoparticles physicochemical characteristics is mandatory as the first step in accurately elucidating their behaviour in different ecosystems. Here in this work, in house-synthesized tellurium-based nanoparticles have been fully characterized for first time by means of a multi-method approach. Once characterized, the effect of these nanoparticles on Staphylococcus aureus and Escherichia coli biofilm biomass and structure was explored and quantified for first time. Moreover, the morphological transformations of tellurium based nanoparticles within the confines of a biofilm are also highlighted. Architectural metric calculations evidenced that nanoparticles were able to reduce the biovolume of the biofilm produced for both bacteria. Interestingly, the interaction between nanoparticles and bacterial communities led to the transformation of telluriumnanoparticles from sphere to rod-shaped nanoparticles. These findings open new insights into the behaviour of a type of uncommon nanoparticles such as tellurium-based nanoparticles on microbial communitiesItem Characterization of AgNPs and AuNPs in sewage sludge by single particle inductively coupled plasma-mass spectrometry(Talanta, 2022) Moreno Martín, Gustavo; Gómez Gómez, Beatriz; León González, María Eugenia De; Madrid Albarrán, María Yolanda; ElsevierThis study develops for the first time an analytical method for the characterization of silver and gold nanoparticles in sewage sludge. The evaluation of the effect of temperature, extracting agent and centrifugation speed and time on the extraction yield was carried out through a multifactorial analysis of variance which allows us to select 289 g, 5 min and 20 mM sodium pyrophosphate tetrabasic as optimal extraction conditions. Under these conditions, the analysis of the extract by single particle inductively coupled plasma-mass spectrometry provided recovery percentages of 70 ± 2% and 56 ± 1% for silver and gold nanoparticles, respectively. Moreover, the complementary results obtained upon analysis of these extracts by transmission electron microscopy and single particle inductively coupled plasma-mass spectrometry showed that the developed method did not modify the original size and shape of these nanoparticles during the extraction procedure. Size detection limits of 23 nm and 16 nm as well as number concentration limits of 3.12 × 109 particles kg−1 and 1.38 × 109 particles kg−1 were obtained for silver and gold nanoparticles, respectively. Moreover, a stability study of silver and gold nanoparticles in sewage sludge for 12 months showed differences between the two nanoparticle types. Although the sizes were not affected during the 12 months, silver nanoparticles underwent an oxidation process from 6 months onwards, which was reflected in an increase in the percentage of ionic silver from 14 ± 1% at 6 months to 24 ± 2% at 12 months. The developed methodology represents a simple, reliable and fast tool for detecting, quantifying and assessing the stability of nanoparticles in an important environmental sample such as sewage sludge.Item Food prospects of selenium enriched-Lactobacillus acidophilus CRL 636 and Lactobacillus reuteri CRL 1101(Journal of Functional Foods, 2017) Pescuma, Micaela; Gómez Gómez, Beatriz; Pérez Corona, María Teresa; Font, Graciela; Madrid Albarrán, María Yolanda; Mozzi, FernandaSelenium, which is present as SeCys in selenoproteins, is involved in cancer prevention, thyroid functioning, and pathogen inhibition. Lactobacilli can biotransform inorganic Se into seleno-amino acids. Growth, Se accumulation and seleno-amino acid formation by Lactobacillus acidophilus CRL636 and L. reuteri CRL1101 in a Se-supplemented medium were studied. Moreover, survival of Se-enriched strains to different pH values and bile salts was analyzed. L. acidophilus CRL636 showed low growth rate in the presence of Se while differences were less evident for L. reuteri CRL1101, which displayed higher amounts of intracellular SeCys and SeMet than the CRL636 strain. Interestingly, both lactobacilli could produce Se-nanoparticles. Se-enriched lactobacilli showed lower growth rates than non-Se exposed cells. The adverse effect of bile salts and the ability to survive at pH 4.0 diminished for the Se-enriched L. reuteri strain. The studied lactobacilli could be used as Se-enriched probiotics or as a vehicle for manufacturing Se-containing fermented foods.Item Using single-particle ICP-MS for unravelling the effect of type of food on the physicochemical properties and gastrointestinal stability of ZnONPs released from packaging materials(Analytica Chimica Acta, 2020) Gómez Gómez, Beatriz; Pérez Corona, María Teresa; Madrid Albarrán, María YolandaThis article discusses the application of Single-Particle Inductively Coupled Plasma Mass Spectrometry (SP-ICP-MS) to study the effect of different types of food (orange juice and chicken breast) on the fate of zinc oxide nanoparticles ZnONPs) migrated from two widely employed food packaging materials (polyethylene terephthalate (PET) and low density polyethylene (LDPE)). The gastrointestinal stability of ZnONPs was also evaluated. The idea behind this study is to track for first time the transformations underwent of nanoparticles in the different steps of their route from packaging to the consumer. The presence of high amount of dissolved zinc in the samples notably influenced size detection limit and the accuracy of SP-ICP-MS measurements. The diameter limits of detection (LODd) were 26 nm, 95 nm, 108 nm and 129 nm for aqueous solution, chicken breast extract and for oral and intestinal extracts, respectively. ZnONPs characterization in juice was not possible with SP-ICP-MS due to nanoparticles size was below LODd. Besides difficulties, SP-ICP-MS after extraction with Tris-HCl allowed us to determine that a 72% of the ZnONPs that migrated to chicken breast were smaller than 95 nm. Complementary to SP-ICP-MS, transmission electron microscopy (TEM) enabled to detect small nanoparticles (<3 nm). The combination of TEM and SP-ICP-MS measurements indicated that nanoparticles in chicken reach the intestine wall as small particles (<10 nm), as aggregates (>200 nm) and as ionic zinc whereas in case of juice only small nanoparticles (<3 nm) and ionic zinc were detected in the intestinal step.Item Availability of zinc from infant formula by in vitro methods (solubility and dialyzability) and size-exclusion chromatography coupled to inductively coupled plasma-mass spectrometry(Journal of Dairy Sciences, 2016) Gómez Gómez, Beatriz; Pérez Corona, María Teresa; Madrid Albarrán, María YolandaZinc bioaccessibility from infant formula was estimated by in vitro methods (solubility and dialyzability) and size-exclusion chromatography (SEC) coupled to inductively coupled plasma-mass spectrometry (ICP-MS). Infant formula samples were first characterized in terms of Zn bound to lipids and proteins and Zn distribution in the aqueous soluble protein fraction. We found that Zn is not incorporated into the lipid fraction of the samples, being mainly associated with the protein fraction (around 100%). Fractionation of Zn-containing proteins in the soluble protein fraction was achieved by SEC-ICP-MS after performing protein extraction with a solution of 100 mM (pH 6.8) Tris-HCl. The percentages of zinc in the soluble protein fraction in the soy-based and lactose-free infant formula were very low, around 7 and 24%, respectively, whereas the content of Zn in the soluble protein fraction of milk-based formula was around 90%. By SEC-ICP-MS, we found that Zn is associated with low-molecular weight compounds (around 10 kDa) in all the infant formulas tested. The percentages of Zn estimated in the in vitro gastrointestinal digests of the infant formula ranged from 30 to 70% and from 1 to 10% for solubility and dialyzability assays, respectively. The dialyzability test resulted in lower than expected scores, as SEC-ICP-MS analysis of the gastrointestinal extracts revealed that Zn is bound to biomolecules with a molecular weight ranging from 1 to 7 kDa, which suggests that dialysis data should be interpreted with caution. Speciation studies are a valuable tool for establishing availability of nutrients and for validating data from dialyzable in vitro methods.Item Silac-based quantitative proteomic analysis of Lactobacillus reuteri CRL 1101 response to the presence of selenite and selenium nanoparticles(Journal of Proteomics, 2019) Gómez Gómez, Beatriz; Pérez Corona, María Teresa; Mozzi, Fernanda; Pescuma, Micaela; Madrid Albarrán, María YolandaStable isotope labeling in cell culture (SILAC) was applied for the first time on a lactic acid bacterium strain (L. reuteri CRL1101) for analyzing differential protein expression associated to selenite(Na2SeO3) and selenium nanoparticles (SeNPs) exposure. 57 and 47 proteins were found de-regulated by >1,5 fold in presence of selenite and SeNPs, respectively. Only 16 out of 104 proteins differentially expressed were commonly altered by selenite and SeNPs. The use of a clustered heat map allows us to visualize relations between the de-regulated proteins and exposure conditions. We identified a number of proteins involved in diverse functions and biological processes such as metabolism of carbohydrates, selenium and lipids; folding, sorting and degradation; environmental information and processing. In presence of both, selenite and SeNPs, proteins related to selenium metabolism such as cystathione beta-lyase and oxidoreductases (thioredoxine reductase and NAD/FAD oxidoreductase) were over expressed. Interestingly, the over expression of thioredoxin reductase could protect the host from oxidizing compounds. An over expression of phage proteins and chaperones with selenite was observed; this result and the fact that a lower cell count was detected when selenite was added could indicate that this latter Se species has a more deleterious effect than the nanoparticles.Item Combined single cell and single particle ICP-TQ-MS analysis to quantitatively evaluate the uptake and biotransformation of tellurium nanoparticles in bacteria(Analytica Chimica Acta, 2020) Gómez Gómez, Beatriz; Corte-Rodríguez, Mario; Pérez Corona, María Teresa; Bettmer, Jörg; Montes-Bayón, María; Madrid Albarrán, María YolandaAssessing the impact of nanoparticles in living systems implies a proper evaluation of their behaviour at single-cell level. Due to the small size of nanoparticles, their accumulation, transformation and location within single cells is challenging. In this work, the combination of single cell/single particle triple quadrupole inductively coupled plasma mass spectrometry (SC/SP-ICP-TQ-MS) analysis along with X-ray diffraction (XRD) and transmission electron microscopy (TEM) measurements has been applied to go deeper into the uptake and biotransformation of tellurium nanoparticles (TeNPs) in two bacterial model organisms, S. aureus and E. coli. The use of SC-ICP-TQ-MS enabled the individual introduction of bacterial cells where tellurium and phosphorous (as constitutive element) were monitored and detected at concentration levels down to femtogram (fg) per cell. S.aureus uptake of TeNPs was 0.5–1.9 fg Te cell−1 and 7–30 fg Te cell−1 in presence of 0.5 and 15 mg Te L−1 of TeNPs, respectively, whereas for E. coli, the amount of Te ranged from 0.08 to 0.88 fg Te cell−1 and from 2 to 36 fg Te cell−1 in presence of 0.5 and 15 mg Te L−1 of TeNPs, respectively. TEM and XRD analysis confirmed the occurrence of TeNPs biotransformation (from nanospheres to nanorods) as the nanoparticles were incorporated into both bacterial strains. Finally, SP-ICP-MS analysis after cell lysis was applied to determine the number of particles/rods per bacteria cell and to perform the dimensional characterization of the rod-shaped TeNPs. The results obtained clearly confirmed high cell-to-cell variability in terms of Te nanorods dimensions and TeNPs uptake. To the best of our knowledge, this is the first time that SC/SP-ICP-TQ-MS along with TEM and XRD analysis have been applied to investigate, quantitatively, nanoparticle uptake in bacterial cells and to estimate the dimensions of biogenic Te nanorods.Item Evaluation of the transformation of selenite and selenium nanoparticles to seleno-amino acids produced by Escherichia coli and Staphylococcus aureus by using liquid chromatography -inductively coupled plasma mass spectrometry and single-particle- inductively coupled plasma mass spectrometry and different sample treatments(Spectrochimica Acta Part B: Atomic Spectroscopy, 2023) Moreno Martín, Gustavo; Espada-Bernabé, Elena; Gómez Gómez, Beatriz; León González, María Eugenia De; Madrid Albarrán, María YolandaDue to the scarce knowledge about the impact of selenium nanoparticles (SeNPs) on bacterial populations, the main objective of this work was focused on evaluating the transformations of SeNPs and selenite in Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). For this purpose, an analytical methodology based on bacteria cell wall disruption, carbamidomethylation, enzymatic hydrolysis and high-performance liquid chromatography couple to inductively coupled plasma mass spectrometry (HPLC-ICP-MS) and HPLC- electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) measurements was developed, the latter for unambiguous identification of selenium species. Once bacteria were cultured in the presence of chitosan modified SeNPs (Ch-SeNPs) and selenite at 0, 1 and 2 mg L 1 Se for 24 h, an enzymatic disruption of the bacterial cell wall using lysozym followed by enzymatic hydrolysis with protease was applied. The use of lysozyme to extract selenium speciesprovided a better efficiency in the total selenium content (higher than 96%), compared to a mechanical disruption of the bacterial cell wall. Analysis of the extracts by anionic exchange HPLC-ICP-MS showed a strong influence of incubation time with protease (24, 48 and 72 h) on selenium chromatographic profile. The results showed that selenocysteine (SeCys) was the only Se species identified in both bacteria representing an 80% of total selenium accumulated. The confirmation of the identity of this Se species was carried out after performing a carbamidomethylation process, prior to enzymatic hydrolysis, and analyzing the extract by reversed phase HPLCESI- MS/MS. These analyses confirmed the presence of SeCys, and no relevant differences were found between the metabolic pathway of both forms of selenium. Moreover, the growth of both bacterial species in the presence of selenite resulted in the formation of biogenic SeNPs. Characterization by TEM before and after their separation from the culture medium showed spherical and monodispersed nanoparticles with an average size (155 ± 19) nm and (172 ± 20) nm for E. coli and S. aureus, respectively. Analysis by spICP-MS showed no significant differences in size with respect to TEM after considering the ionization efficiency of Se.Item Selenium health benefit values and Hg and Se speciation studies for elucidating the quality and safety of highly consumed fish species and fish-derived products(Food Chemistry, 2024) Fernández Bautista, Tamara; Gómez Gómez, Beatriz; Gracia Lor, Emma; Pérez Corona, María Teresa; Madrid Albarrán, María YolandaNowadays, there has been an increase in the consumption of fish-derived products that constitute alternative ways of introducing fish into the diet, such as fish roe and products made from fish (crab sticks and surimiderived products). However, there is no data available considering selenium and mercury total contents along with speciation studies in these kinds of samples, which are mandatory for a proper safety and quality assessment. In this study, Selenium Health Benefit Values (HBVSe) in fish and fish-derived products were evaluated, resulting in positive in all cases. Selenium speciation studies performed by HPLC-ICP-MS and confirmed by HPLC-ESI-MS/MS revealed that SeMet and SeMeSeCys are present in fish and fish-derived products. Finally, Hg speciation studies show that the percentage of Hg2+ increases in fishes lower in the food chain as well as in fishderived products.