Antonio L. Crego

Universidad de Alcalá

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Marı́a Luisa Marina

Universidad de Alcalá

Carmen García‐Ruiz

Universidad de Alcalá

María Castro‐Puyana

Universidad de Alcalá

Laura Sánchez‐Hernández

Instituto Politécnico Nacional

Elena Domínguez‐Vega

Leiden University Medical Center

Elena Sánchez‐López

Leiden University Medical Center

Miguel Guijarro‐Díez

Universidad de Alcalá

Leonor Nozal

Universidad de Alcalá

Bezhan Chankvetadze

Tbilisi State University

Ana Belén Martínez‐Girón

Universidad de Alcalá

Cooperative Institutions

Universidad de Alcalá

Consejo Superior de Investigaciones Científicas

National University of Distance Education

Universidad Autónoma de Madrid

Leiden University Medical Center

Universidad Complutense de Madrid

Coe College

Instituto de Fermentaciones Industriales

IMDEA Water

Netherlands Metabolomics Centre

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Metabolic fingerprinting of chemotherapy-resistant prostate cancer stem cells. An untargeted metabolomic approach by liquid chromatography-mass spectrometry

Frontiers in Cell and Developmental Biology (2022)

Alicia Bort Belén G. Sánchez Carlos León Leonor Nozal José María Mora-Rodríguez

Chemoresistance is one of the most important challenges in cancer therapy. The presence of cancer stem cells within the tumor may contribute to chemotherapy resistance since these cells express high levels of extrusion pumps and xenobiotic metabolizing enzymes that inactivate the therapeutic drug. Despite the recent advances in cancer cell metabolism adaptations, little is known about the metabolic adaptations of the cancer stem cells resistant to chemotherapy. In this study, we have undertaken an untargeted metabolomic analysis by liquid chromatography-high-resolution spectrometry combined with cytotoxicity assay, western blot, quantitative real-time polymerase chain reaction (qPCR), and fatty acid oxidation in a prostate cancer cell line resistant to the antiandrogen 2-hydroxiflutamide with features of cancer stem cells, compared to its parental androgen-sensitive cell line. Metabolic fingerprinting revealed 106 out of the 850 metabolites in ESI+ and 67 out of 446 in ESI- with significant differences between the sensitive and the resistant cell lines. Pathway analysis performed with the unequivocally identified metabolites, revealed changes in pathways involved in energy metabolism as well as posttranscriptional regulation. Validation by enzyme expression analysis indicated that the chemotherapy-resistant prostate cancer stem cells were metabolically dormant with decreased fatty acid oxidation, methionine metabolism and ADP-ribosylation. Our results shed light on the pathways underlying the entry of cancer cells into dormancy that might contribute to the mechanisms of drug resistance.

10.3389/fcell.2022.1005675

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Citations (4)

Chiral separation of a basic drug with two chiral centers by electrokinetic chromatography for its pharmaceutical development

Journal of Chromatography A (2016)

Ana Belén Martínez‐Girón Marı́a Luisa Marina Antonio L. Crego

Micellar electrokinetic chromatography

Chiral resolution

10.1016/j.chroma.2016.08.041

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Citations (17)

Separation of enantiomers of norephedrine by capillary electrophoresis using cyclodextrins as chiral selectors: Comparative CE and NMR studies

Electrophoresis (2012)

Ketevan Lomsadze Elena Domínguez‐Vega Antonio Salgado Antonio L. Crego Gerhard K. E. Scriba

In this study, the enantiomer migration order ( EMO ) of norephedrine ( NEP ) in the presence of various CD s was investigated by CE . NMR and CE techniques were used to analyze the mechanism of the chiral recognition between NEP enantiomers and four CD s, i.e., native α‐ CD , β‐ CD , heptakis(2,3‐di‐O‐acetyl‐6‐O‐sulfo)‐β‐ CD ( HDAS ‐β‐ CD ), and heptakis(2,3‐di‐O‐methyl‐6‐O‐sulfo)‐β‐ CD (HDMS‐β‐ CD ). EMO was reversed in the presence of α‐ CD and β‐ CD , although only minor differences in the structures of the complexes formed between NEP and these CD s could be derived from rotating frame nuclear Overhauser experiments ( ROESY ). The complexes between the enantiomers of NEP and the sulfated CD s, HDMS‐β‐ CD , and HDAS ‐β‐ CD , were substantially different. However, EMO of NEP was identical in the presence of these CD s. HDAS ‐β‐ CD proved to be the most suitable chiral selector for the CE enantioseparation of NEP .

Chirality

Proton NMR

10.1002/elps.201200062

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Citations (46)

Enantiomeric separation of chiral polycyclic musks by capillary electrophoresis: Application to the analysis of cosmetic samples

Journal of Chromatography A (2009)

Ana Belén Martínez‐Girón Antonio L. Crego María José González Marı́a Luisa Marina

Matrix (chemical analysis)

10.1016/j.chroma.2009.12.021

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Citations (45)

A simple capillary gel electrophoresis approach for efficient and reproducible DNA separations. Analysis of genetically modified soy and maize

Journal of Separation Science (2007)

Laura M. Sanchez Ramón González Antonio L. Crego Alejandro Cifuentes

Abstract It is generally assumed that in order to achieve suitable separations of DNA fragments, capillary gel electrophoresis (CGE)‐coated capillaries should be used. In this work, a new method is presented that allows to obtain reproducible CGE separations of DNA fragments using bare fused‐silica capillaries without any previous coating step. The proposed method only requires: (i) a capillary washing with 0.1 M hydrochloric acid between injections and (ii) a running buffer composed of Tris‐phosphate‐ethylenediamine tetraacetic acid (EDTA) and 4.5% of 2‐hydroxyethyl cellulose (HEC) as sieving polymer. The use of this new CGE procedure gives highly resolved and reproducible separations of DNA fragments ranging from 50 to 750 bp. The separation of these DNA fragments is accomplished in less than 30 min with efficiencies up to 1.7×10 6 plates/m. Reproducibility values of migration times (given as %RSD) for the analyzed DNA fragments are better than 1.0% ( n = 4) for the same day, 2.2% ( n = 16) for four different days, and 2.3% ( n = 16) for four different capillaries. The usefulness of this separation method is demonstrated by detecting genetically modified maize and genetically modified soy after DNA amplification by PCR. This new CGE procedure together with LIF as detector provides sensitive analysis of 0.9% of Bt11 maize, Mon810 maize, and Roundup Ready soy in flours with S/N up to 542. These results demonstrate the usefulness of this procedure to fulfill the European regulation on detection of genetically modified organisms in foods.

Genetically modified maize

Hydroxyethyl cellulose

10.1002/jssc.200600195

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Citations (23)

Potential of vancomycin for the enantiomeric resolution of FMOC‐amino acids by capillary electrophoresis‐ion‐trap‐mass spectrometry

Electrophoresis (2013)

Laura Sánchez‐Hernández Elena Domínguez‐Vega Cristina Montealegre María Castro‐Puyana Marı́a Luisa Marina

The potential of the antibiotic vancomycin (VC) as chiral selector for the enantiomeric separation of amino acids by CE-ESI-MS/MS² was investigated for the first time in this work. Derivatization of amino acids with FMOC-Cl was carried out to enable their interaction with VC as well as the formation of precursor ions with larger m/z which were employed in MS² experiments. The partial filling of a coated capillary was employed to avoid the loss in MS sensitivity originated by the introduction of VC in the ionization source. Under optimized conditions, the simultaneous enantiomeric separation and unequivocal identification of 17 amino acids (two of them being nonprotein amino acids) took place in about 20 min with LODs in the micromolar range.

Capillary electrophoresis–mass spectrometry

10.1002/elps.201300489

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Citations (44)

Properties and Structure of Glassy TeO2 and Binary Potassium and Boron Tellurites

Journal of Undergraduate Reports in Physics (2018)

Brittney M. Hauke Emma R. Barney Antonio L. Crego Gabriella Tarantino Mario Affatigato

Tellurite glasses show potential for use in mid-infrared optical applications1, but their structure has not been intensively studied. While they do not conduct light better than chalcogenides, which are currently the best glasses for infrared optics, they are much easier to produce. Potassium and boron tellurite glasses, including single component, rapidly cooled TeO2, are reported and studied here. The results include the Glass Transition Temperature (Tg) measurements and Raman spectra. Proposed structural models are also discussed.

Potassium silicate

Component (thermodynamics)

10.1063/1.5109555

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Citations (3)

Chiral Separations by Capillary Electrophoresis

Elena Sánchez‐López María Castro‐Puyana Marı́a Luisa Marina Antonio L. Crego

Abstract The enantiomeric separation of chiral compounds is nowadays a promising topic in analytical sciences. CE is considered as a well‐established tool to perform chiral separations of a wide range of analytes in different research fields such as pharmaceutical, food and chemical industries, and clinical and environmental laboratories. Due to its numerous advantages such as high efficiency, simplicity, and low consumption of reagents, chiral selectors, and samples, CE plays an important role in the separation of enantiomers. Advantages and limitations of the different CE modes are displayed in the present chapter, discussing their possibilities and the actual trends. Hyphenation with MS detection is also discussed, highlighting the potential of this coupling as well as its weaknesses and, the most commonly used strategies to minimize its drawbacks. Most recent and relevant applications of chiral CE are summarized and illustrated with tables including the analysis of pharmaceutical, biological, and environmental samples, food and beverages, and agrochemicals. Finally, concluding remarks and future trends are also included describing the actual state of the art and challenges of chiral CE techniques.

Simplicity

10.1002/9783527678129.assep043

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Citations (23)

Recent contributions of capillary electrophoresis to neuroscience

TrAC Trends in Analytical Chemistry (2015)

Elena Sánchez‐López Cristina Montealegre Antonio L. Crego Marı́a Luisa Marina

10.1016/j.trac.2014.12.008

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Citations (21)

Recent contributions for improving sensitivity in chiral CE

Electrophoresis (2017)

Antonio L. Crego María Luisa Mateos Leonor Nozal

Abstract The flexibility and versatility of the chiral CE are unrivaled and the same instrumentation can be used to separate a diverse range of analytes, both large and small molecules, whether charged or uncharged. However, one of the disadvantages is generally thought to be the poor sensitivity of ultraviolet (UV) detection, which is the most popular among CE detectors. This review focuses on methodologies and applications regarding improvements of sensitivity in chiral CE published in the last 2 years (June 2015 until May 2017). This contribution continues to update this series of biannual reviews, first published in Electrophoresis in 2006. The main body of the review brings a survey of publications organized according to different approaches to detect a low amount of analytes, either by sample treatment procedures or by in‐capillary sample preconcentration techniques, both using UV detection, or even by employing detection systems more sensitive than UV absorption, such as LIF or MS. This review provides comprehensive tables listing the new approaches in sensitive chiral CE with categorizing by the fundamental mechanism to enhance the sensitivity, which provide relevant information on the strategies employed. The concluding remarks in the final part of the review evaluate present state of art and the trends for sensitivity enhancement in chiral CE.

Instrumentation

10.1002/elps.201700293

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Citations (13)