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1.
Analyst ; 148(16): 3883-3891, 2023 Aug 07.
Artículo en Inglés | MEDLINE | ID: mdl-37458061

RESUMEN

Metabolites in biological matrices belong to diverse chemical groups, ranging from non-polar long-chain fatty acids to small polar molecules. The goal of untargeted metabolomic analysis is to measure the highest number of metabolites in the sample. Nevertheless, from an analytical point of view, no single technique can measure such a broad spectrum of analytes. Therefore, we selected a method based on GC-MS and LC-MS with two types of stationary phases for the untargeted profiling of gastrointestinal stromal tumours. The procedure was applied to GIST xenograft samples (n = 71) representing four different mutation models, half of which were treated with imatinib. We aimed to verify the method coverage and advantages of applying each technique. RP-LC-MS measured most metabolites due to a significant fraction of lipid components of the tumour tissue. What is unique and worth noting is that all applied techniques were able to distinguish between different mutation models. However, for detecting imatinib-induced alterations in the GIST metabolome, RP-LC-MS and GC-MS proved to be more relevant than HILIC-LC-MS, resulting in a higher number of significantly changed metabolites in four treated models. Undoubtedly, the inclusion of all mentioned techniques makes the method more comprehensive. Nonetheless, for green chemistry and time and labour saving, we assume that RP-LC-MS and GC-MS analyses are sufficient to cover the global GIST metabolome.


Asunto(s)
Tumores del Estroma Gastrointestinal , Humanos , Tumores del Estroma Gastrointestinal/tratamiento farmacológico , Tumores del Estroma Gastrointestinal/genética , Xenoinjertos , Mesilato de Imatinib/farmacología , Mesilato de Imatinib/uso terapéutico , Metaboloma , Metabolómica/métodos , Mutación
2.
Anal Bioanal Chem ; 415(22): 5511-5528, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37460824

RESUMEN

In the present study, we developed and validated a fast, simple, and sensitive quantitative method for the simultaneous determination of eleven nucleosides and deoxynucleosides from urine samples. The analyses were performed with the use of liquid chromatography coupled with triple quadrupole mass spectrometry. The sample pretreatment procedure was limited to centrifugation, vortex mixing of urine samples with a methanol/water solution (1:1, v/v), evaporation and dissolution steps. The analysis lasted 20 min and was performed in dynamic multiple reaction monitoring mode (dMRM) in positive polarity. Process validation was conducted to determine the linearity, precision, accuracy, limit of quantification, stability, recovery and matrix effect. All validation procedures were carried out in accordance with current FDA and EMA regulations. The validated method was applied for the analysis of 133 urine samples derived from bladder cancer patients before tumor resection and 24 h, 2 weeks, and 3, 6, 9, and 12 months after the surgery. The obtained data sets were analyzed using a linear mixed-effect model. The analysis revealed that concentration level of 2-methylthioadenosine was decreased, while for inosine, it was increased 24 h after tumor resection in comparison to the preoperative state. The presented quantitative longitudinal study of urine nucleosides and deoxynucleosides before and up to 12 months after bladder tumor resection brings additional prospective insight into the metabolite excretion pattern in bladder cancer disease. Moreover, incurred sample reanalysis was performed proving the robustness and repeatability of the developed targeted method.


Asunto(s)
Nucleósidos , Neoplasias de la Vejiga Urinaria , Humanos , Nucleósidos/análisis , Estudios Longitudinales , Espectrometría de Masas en Tándem/métodos , Neoplasias de la Vejiga Urinaria/cirugía , Metabolómica , Cromatografía Líquida de Alta Presión/métodos
3.
Anal Chem ; 94(31): 11070-11080, 2022 08 09.
Artículo en Inglés | MEDLINE | ID: mdl-35903961

RESUMEN

Large datasets of chromatographic retention times are relatively easy to collect. This statement is particularly true when mixtures of compounds are analyzed under a series of gradient conditions using chromatographic techniques coupled with mass spectrometry detection. Such datasets carry much information about chromatographic retention that, if extracted, can provide useful predictive information. In this work, we proposed a mechanistic model that jointly explains the relationship between pH, organic modifier type, temperature, gradient duration, and analyte retention based on liquid chromatography retention data collected for 187 small molecules. The model was built utilizing a Bayesian multilevel framework. The model assumes (i) a deterministic Neue equation that describes the relationship between retention time and analyte-specific and instrument-specific parameters, (ii) the relationship between analyte-specific descriptors (log P, pKa, and functional groups) and analyte-specific chromatographic parameters, and (iii) stochastic components of between-analyte and residual variability. The model utilizes prior knowledge about model parameters to regularize predictions which is important as there is ample information about the retention behavior of analytes in various stationary phases in the literature. The usefulness of the proposed model in providing interpretable summaries of complex data and in decision making is discussed.


Asunto(s)
Cromatografía Líquida de Alta Presión , Teorema de Bayes , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Liquida/métodos , Espectrometría de Masas
4.
Molecules ; 26(8)2021 Apr 11.
Artículo en Inglés | MEDLINE | ID: mdl-33920347

RESUMEN

Bladder cancer (BC) is a common malignancy of the urinary system and a leading cause of death worldwide. In this work, untargeted metabolomic profiling of biological fluids is presented as a non-invasive tool for bladder cancer biomarker discovery as a first step towards developing superior methods for detection, treatment, and prevention well as to further our current understanding of this disease. In this study, urine samples from 24 healthy volunteers and 24 BC patients were subjected to metabolomic profiling using high throughput solid-phase microextraction (SPME) in thin-film format and reversed-phase high-performance liquid chromatography coupled with a Q Exactive Focus Orbitrap mass spectrometer. The chemometric analysis enabled the selection of metabolites contributing to the observed separation of BC patients from the control group. Relevant differences were demonstrated for phenylalanine metabolism compounds, i.e., benzoic acid, hippuric acid, and 4-hydroxycinnamic acid. Furthermore, compounds involved in the metabolism of histidine, beta-alanine, and glycerophospholipids were also identified. Thin-film SPME can be efficiently used as an alternative approach to other traditional urine sample preparation methods, demonstrating the SPME technique as a simple and efficient tool for urinary metabolomics research. Moreover, this study's results may support a better understanding of bladder cancer development and progression mechanisms.


Asunto(s)
Metaboloma , Metabolómica/métodos , Neoplasias de la Vejiga Urinaria/orina , Anciano , Ácido Benzoico/orina , Estudios de Casos y Controles , Cromatografía Liquida , Ácidos Cumáricos/orina , Femenino , Glicerofosfolípidos/orina , Hipuratos/orina , Histidina/orina , Humanos , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Fenilalanina/metabolismo , Microextracción en Fase Sólida/métodos , Espectrometría de Masas en Tándem , Vejiga Urinaria/metabolismo , Vejiga Urinaria/patología , Neoplasias de la Vejiga Urinaria/diagnóstico , Neoplasias de la Vejiga Urinaria/patología , beta-Alanina/orina
5.
Molecules ; 24(6)2019 Mar 24.
Artículo en Inglés | MEDLINE | ID: mdl-30909656

RESUMEN

Urinary pterins have been found as potential biomarkers in many pathophysiological conditions including inflammation, viral infections, and cancer. However, pterins determination in biological samples is difficult due to their degradation under exposure to air, light, and heat. Besides, they occur at shallow concentration levels, and thus, standard UV detectors cannot be used without additional sample preconcentration. On the other hand, ultra-sensitive laser-induced fluorescence (LIF) detection can be used since pterins exhibit native fluorescence. The main factor that limits an everyday use of LIF detectors is its high price. Here, an alternative detector, i.e., light-emitted diode induced fluorescence (LEDIF) detector, was evaluated for the determination of pterins in urine samples after capillary electrophoresis (CE) separation. An optimized method was validated in terms of linearity range, limit of detection (LOD), limit of quantification (LOQ), intra- and interday precision and accuracy, sample stability in the autosampler, and sample stability during the freezing/thawing cycle. The obtained LOD (0.1 µM) and LOQ (0.3 µM) values were three-order of magnitude lower compared to UV detector, and two orders of magnitude higher compared to previously reported house-built LIF detector. The applicability of the validated method was demonstrated in the analysis of urine samples from healthy individuals and cancer patients.


Asunto(s)
Biomarcadores de Tumor/orina , Pterinas/orina , Estudios de Casos y Controles , Electroforesis Capilar , Humanos , Límite de Detección , Oxidación-Reducción , Espectrometría de Fluorescencia/instrumentación , Neoplasias Urológicas/diagnóstico
6.
Rapid Commun Mass Spectrom ; 32(11): 919-927, 2018 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-29578620

RESUMEN

RATIONALE: Tocopherols and tocotrienols are chemical compounds insusceptible to the ionization process under atmospheric pressure conditions. Therefore, the selection of the optimal ion source settings for their quantification requires special attention. The aim of this study was to analyse the influence of the APCI source parameters on the response of tocochromanols and two related compounds. METHODS: Standard solutions of target compounds were injected on the high-performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry (HPLC/APCI-MS/MS) system separately and analysed with 30 randomly selected ion source settings. The obtained responses were modelled by multivariate linear regression with least absolute shrinkage and selection operator. The developed models were used to choose the best APCI conditions. RESULTS: Multivariate linear models were built for eight tocochromanols, trolox and BHT. The APCI settings derived from the models did not increase the peak areas obtained for T and T3 during the ionization process. Ionization conditions based on models for trolox and BHT improved analytical responses by 12-36% and 4-32%, respectively. The application of the ion source settings optimal for trolox and BHT to tocochromanols did not result in better analytical responses. CONCLUSIONS: The ionization pattern of tocochromanols in the APCI source is problematic and should be further investigated. Modelling methodology for response improvement presented in this study can be applied in similar studies.

7.
Anal Chem ; 89(24): 13422-13428, 2017 12 19.
Artículo en Inglés | MEDLINE | ID: mdl-29172453

RESUMEN

A stacking approach in capillary electrophoresis based on the reversal of the analytes' effective electrophoretic velocities at a dynamic stacking boundary formed between charged micelles (i.e., from long chain ionic surfactants) and neutral cyclodextrins (i.e., native α-, ß-, or γ-cyclodextrin) is presented. The approach was demonstrated by the long injection of samples in a micellar solution followed by injection of a cyclodextrin solution zone, and then separation by co-electro-osmotic flow capillary zone electrophoresis. The reversal is caused by the formation of stable cyclodextrin-surfactant complexes at the boundary that significantly decreased the retention factor of the analytes in the presence of a micellar pseudostationary phase. The dynamic boundary was formed at the cyclodextrin zone as the micelles penetrated this zone. Under optimum conditions, the boundary disappears, and the stacking ends when all the micelles have electrophoretically migrated to the boundary. Cationic and anionic small molecules were enriched using oppositely charged micelles from sodium dodecyl sulfate and cetyltrimethylammonium bromide, respectively. There were 1-2 orders of concentration magnitude improvement in analyte detection, which is expected in stacking with hydrodynamic injection. The improvements in the peak signals (height/corrected area) were up to 236/445 and 101/76 for the cationic and anionic analytes tested, respectively. Linearity (r2) and repeatability (%RSD of migration time, peak height, and corrected peak area) under the chosen stacking conditions (cations/anions) were ≥0.998/≥0.995 and ≤3.8%/≤5.7%, respectively. The stacking approach was also implemented in the direct analysis of peptides from trypsin digested bovine serum albumin.

8.
Electrophoresis ; 38(13-14): 1730-1735, 2017 07.
Artículo en Inglés | MEDLINE | ID: mdl-28342262

RESUMEN

A significant shift of migration time of nonretained compounds (ascorbic acid and cysteine) in micellar electrokinetic chromatography was observed under variation of sample matrix composition. The shift was affected by borate buffer concentration in sample matrix, sample injection time, and pH of BGE (80 mM SDS, Tris/HCl). Surprisingly, longer migration time of analyte was recorded at higher pH of separation buffer. These observations were linked to transient isotachophoresis process. Computer simulation with Simul5 software was conducted to support this hypothesis. The manuscript documents rarely reported in the literature phenomenon of isotachophoresis in micellar electrokinetic chromatography. The analytical potential of described observations was also discussed.


Asunto(s)
Electrocromatografía Capilar/métodos , Cromatografía Capilar Electrocinética Micelar/métodos , Isotacoforesis/métodos , Ácido Ascórbico , Cisteína , Concentración de Iones de Hidrógeno , Modelos Químicos , Reproducibilidad de los Resultados , Programas Informáticos , Factores de Tiempo
9.
Electrophoresis ; 36(1): 135-43, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25255727

RESUMEN

Multidimensional separation where two or more orthogonal displacement mechanisms are combined is a promising approach to increase peak capacity in CE. The combinations allow dramatic improvement of analytical performance since the total peak capacity is given by a product of the peak capacities of all methods. The initial reports were concentrated on the construction of effective connections between capillaries for 2D analysis. Today, 2D and 3D CE systems are now able to separate real complex biological or environmental mixtures with good repeatability, improved resolution with minimal loss of sample. This review will present the developments in the field of multidimensional CE during the last 15 years. The endeavors in this specific field were on the development of interfaces, interface-free techniques including integrated separations, microdevices, and on-line sample concentration techniques to improve detection sensitivity.


Asunto(s)
Electroforesis Capilar/instrumentación , Animales , Diálisis , Electroforesis Capilar/métodos , Diseño de Equipo , Humanos
10.
J AOAC Int ; 98(2): 345-53, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25905740

RESUMEN

The phenanthrene skeleton is an important moiety in medical chemistry as it is present in steroidal drugs used as anti-inflammatory and anti-asthmatic agents as well as synthetic hormones or potassium sparing diuretics. Chromatographic properties of 14 derivatives containing the phenanthrene skeleton in their structure with known lipophilicity have been studied. NP, RP, and cyano-bonded silica stationary phases with three binary mobile phases (acetonitrile-water, acetone-water, and acetone-petroleum ether) were tested. Obtained chromatographic data were correlated with the lipophilicity expressed as values of log partition coefficient (P). The presented study was undertaken to find the best TLC system and chromatographic data processing method in order to predict log P values. Correlations between chromatographic data and measurements of lipophilicity of compounds were presented as results of established quantitative structure-retention relationships. Principal component analysis and cluster analysis were used to investigate the similarities among chromatographic systems.


Asunto(s)
Cromatografía en Capa Delgada/métodos , Interpretación Estadística de Datos , Estudios de Evaluación como Asunto , Fenantrenos/química , Relación Estructura-Actividad Cuantitativa
11.
Biomed Chromatogr ; 28(6): 802-9, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24861747

RESUMEN

The aim of the study was to develop and validate a simple and rapid method for the determination of cyclosporine A (CsA) in ocular rabbit tissues using reversed-phase ultra-high-performance liquid chromatography (UHPLC) with UV detection. Previous publications on chromatographic methods of CsA determination in ocular tissues involved only reversed-phase HPLC separation, usually in combination with such detection techniques as radio-immunoassay and mass spectrometry. The application of the UHPLC technique allowed us to significantly decrease the analysis time. Cyclosporine D (CsD) was applied as the internal standard. Satisfactory separation was achieved on an XB-C18 Kinetex column at 60°C with the use of gradient elution mode. The retention times of CsA and CsD were found to be 4.5 and 5.1 min, respectively. The developed assay is specific, sensitive (limit of detection = 6 ng/mL and limit of quantitation = 18 ng/mL) and linear within the analyte concentration range of 0.018-5 µg/mL, with a correlation coefficient of 0.999. High sensitivity, low injection volume (10 µL), short time of analysis (6.5 min) and simplicity make this method useful for the fast analysis of CsA in rabbit ocular tissues and fluids: lacrimal fluid, aqueous humor, cornea, conjunctiva and eye globe.


Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Ciclosporina/análisis , Inmunosupresores/análisis , Animales , Humor Acuoso/química , Cromatografía de Fase Inversa/métodos , Ojo/química , Aparato Lagrimal/química , Masculino , Conejos
12.
Sci Rep ; 14(1): 18854, 2024 08 14.
Artículo en Inglés | MEDLINE | ID: mdl-39143107

RESUMEN

The rapid and sensitive indicator of inflammation in the human body is C-Reactive Protein (CRP). Determination of CRP level is important in medical diagnostics because, depending on that factor, it may indicate, e.g., the occurrence of inflammation of various origins, oncological, cardiovascular, bacterial or viral events. In this study, we describe an interferometric sensor able to detect the CRP level for distinguishing between no-inflammation and inflammation states. The measurement head was made of a single mode optical fiber with a microsphere structure created at the tip. Its surface has been biofunctionalized for specific CRP bonding. Standardized CRP solutions were measured in the range of 1.9 µg/L to 333 mg/L and classified in the initial phase of the study. The real samples obtained from hospitalized patients with diagnosed Urinary Tract Infection or Urosepsis were then investigated. 27 machine learning classifiers were tested for labeling the phantom samples as normal or high CRP levels. With the use of the ExtraTreesClassifier we obtained an accuracy of 95% for the validation dataset. The results of real samples classification showed up to 100% accuracy for the validation dataset using XGB classifier.


Asunto(s)
Proteína C-Reactiva , Aprendizaje Automático , Humanos , Proteína C-Reactiva/análisis , Infecciones Urinarias/diagnóstico , Infecciones Urinarias/orina , Interferometría/métodos , Inflamación/diagnóstico , Inflamación/orina , Sepsis/diagnóstico , Sepsis/orina , Técnicas Biosensibles/métodos , Fibras Ópticas
13.
Int J Nanomedicine ; 19: 3973-3989, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38711615

RESUMEN

Graphene and graphene-based materials have attracted growing interest for potential applications in medicine because of their good biocompatibility, cargo capability and possible surface functionalizations. In parallel, prototypic graphene-based devices have been developed to diagnose, imaging and track tumor growth in cancer patients. There is a growing number of reports on the use of graphene and its functionalized derivatives in the design of innovative drugs delivery systems, photothermal and photodynamic cancer therapy, and as a platform to combine multiple therapies. The aim of this review is to introduce the latest scientific achievements in the field of innovative composite graphene materials as potentially applied in cancer therapy. The "Technology and Innovation Roadmap" published in the Graphene Flagship indicates, that the first anti-cancer drugs using graphene and graphene-derived materials will have appeared on the market by 2030. However, it is necessary to broaden understanding of graphene-based material interactions with cellular metabolism and signaling at the functional level, as well as toxicity. The main aspects of further research should elucidate how treatment methods (e.g., photothermal therapy, photodynamic therapy, combination therapy) and the physicochemical properties of graphene materials influence their ability to modulate autophagy and kill cancer cells. Interestingly, recent scientific reports also prove that graphene nanocomposites modulate cancer cell death by inducing precise autophagy dysfunctions caused by lysosome damage. It turns out as well that developing photothermal oncological treatments, it should be taken into account that near-infrared-II radiation (1000-1500 nm) is a better option than NIR-I (750-1000 nm) because it can penetrate deeper into tissues due to less scattering at longer wavelengths radiation.


Asunto(s)
Antineoplásicos , Grafito , Neoplasias , Grafito/química , Humanos , Antineoplásicos/química , Antineoplásicos/farmacología , Neoplasias/tratamiento farmacológico , Sistemas de Liberación de Medicamentos/métodos , Fotoquimioterapia/métodos , Autofagia/efectos de los fármacos , Animales , Nanocompuestos/química , Nanocompuestos/uso terapéutico , Nanomedicina
14.
Sci Rep ; 13(1): 15280, 2023 09 15.
Artículo en Inglés | MEDLINE | ID: mdl-37714912

RESUMEN

Pulmonary arterial hypertension is a rare but life-threatening and clinically heterogeneous disease. The diagnostic schedule of this disorder is complex, and no specific indicator of the arterial etiology has been explored. In this study, untargeted plasma metabolomics was applied to evaluate the metabolic fingerprints of pulmonary arterial hypertension patients. Plasma samples were prepared using a new approach, which applies proteinase K during the sample preparation procedure to increase the metabolite coverage. The metabolic fingerprints were determined via LC-MS and subsequently analyzed with the use of both uni- and multivariate statistics. A total of 21 metabolites were discovered to be significantly altered in pulmonary arterial hypertensive patients. The metabolites were mainly related to the phospholipid metabolic pathways. In this study, decreases were found in the phosphatidylcholines (PCs) [PC(32:0), PC(40:7), PC(42:7)], phosphatidylethanolamine PE(18:0/18:2), lysophosphatidylethanolamines (LPEs) [LPE(22:6), LPE(18:2), LPE(18:0), LPE(20:4), LPE(20:1), LPE(20:0)], lysophosphatidylcholine LPC(20:4) and lysophosphatidylserine LPS(19:0), as well as increase of sphingomyelin SM(36:2), in the plasma samples of pulmonary arterial hypertensive patients in comparison to the control group. Besides their function as components of the biological membranes, these metabolites are also involved in the intracellular signaling pathways that are related to cell proliferation and apoptosis. The results obtained during this study confirm the potential of (untargeted) metabolomics to identify the molecular characteristics of the pathophysiology of pulmonary arterial hypertension. The clinical relevance of this study constitutes the selection of a metabolic panel that can potentially detect and properly diagnose the disease.


Asunto(s)
Hipertensión Pulmonar , Hipertensión Arterial Pulmonar , Humanos , Endopeptidasa K , Hipertensión Pulmonar Primaria Familiar , Metabolómica , Arteria Pulmonar
15.
J Biophotonics ; 16(9): e202300095, 2023 09.
Artículo en Inglés | MEDLINE | ID: mdl-37285226

RESUMEN

The study presents an optical method supported by machine learning for discriminating urinary tract infections from an infection capable of causing urosepsis. The method comprises spectra of spectroscopy measurement of artificial urine samples with bacteria from solid cultures of clinical E. coli strains. To provide a reliable classification of results assistance of 27 algorithms was tested. We proved that is possible to obtain up to 97% accuracy of the measurement method with the use of use of machine learning. The method was validated on urine samples from 241 patients. The advantages of the proposed solution are the simplicity of the sensor, mobility, versatility, and low cost of the test.


Asunto(s)
Sepsis , Infecciones Urinarias , Humanos , Escherichia coli , Infecciones Urinarias/diagnóstico , Sepsis/diagnóstico , Sepsis/etiología , Aprendizaje Automático , Medición de Riesgo
16.
J Pharm Biomed Anal ; 235: 115602, 2023 Oct 25.
Artículo en Inglés | MEDLINE | ID: mdl-37536113

RESUMEN

Despite a large number of studies, the pathogenesis of polycystic ovary syndrome (PCOS) still remains unexplained. In light of ambiguous observations reported in metabolomics, there is a need to carry out studies focusing on confirming the discriminating power of the proposed metabolomics biomarkers. Our research aimed to perform a validation study of metabolites detected in our previous study from serum samples, on the new set of samples obtained from PCOS women and healthy controls to confirm previously selected compounds. Additionally, the second biological matrix - urine - was used to get a more comprehensive insight into metabolic alterations. We applied two analytical techniques - gas chromatography and liquid chromatography coupled with mass spectrometry to analyze both serum and urine samples obtained from 35 PCOS patients and 35 healthy women. Thank to our approach, we identified and described a comprehensive set of metabolites altered in PCOS patients. Results of our study indicate increased steroid hormone synthesis, alteration in sphingo- and phospholipids metabolism, and disturbed fatty acids metabolism. Moreover, the citric acid cycle, γ-glutamyl cycle, vitamin B metabolism, and a few primary amino acids like tryptophan, phenylalanine, histidine, and alanine are altered.


Asunto(s)
Síndrome del Ovario Poliquístico , Humanos , Femenino , Cromatografía de Gases y Espectrometría de Masas/métodos , Metabolómica/métodos , Espectrometría de Masas/métodos , Cromatografía Liquida/métodos
17.
Transl Oncol ; 30: 101632, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-36774883

RESUMEN

BACKGROUND: Although imatinib is a well-established first-line drug for treating a vast majority of gastrointestinal stromal tumours (GIST), GISTs acquire secondary resistance during therapy. Multi-omics approaches provide an integrated perspective to empower the development of personalised therapies through a better understanding of functional biology underlying the disease and molecular-driven selection of the best-targeted individualised therapy. In this study, we applied integrative metabolomic and transcriptomic analyses to elucidate tumour biochemical processes affected by imatinib treatment. MATERIALS AND METHODS: A GIST xenograft mouse model was used in the study, including 10 mice treated with imatinib and 10 non-treated controls. Metabolites in tumour extracts were analysed using gas chromatography coupled with mass spectrometry (GC-MS). RNA sequencing was also performed on the samples subset (n=6). RESULTS: Metabolomic analysis revealed 21 differentiating metabolites, whereas next-generation RNA sequencing data analysis resulted in 531 differentially expressed genes. Imatinib significantly changed the profile of metabolites associated mainly with purine and pyrimidine metabolism, butanoate metabolism, as well as alanine, aspartate, and glutamate metabolism. The related changes in transcriptomic profiles included genes involved in kinase activity and immune responses, as well as supported its impact on the purine biosynthesis pathway. CONCLUSIONS: Our multi-omics study confirmed previously known pathways involved in imatinib anticancer activity as well as correlated imatinib-relevant downregulation of expression of purine biosynthesis pathway genes with the reduction of respectful metabolites. Furthermore, considering the importance of the purine biosynthesis pathway for cancer proliferation, we identified a potentially novel mechanism for the anti-tumour activity of imatinib. Based on the results, we hypothesise metabolic modulations aiming at the reduction in purine and pyrimidine pool may ensure higher imatinib efficacy or re-sensitise imatinib-resistant tumours.

18.
Pharmaceuticals (Basel) ; 16(12)2023 Dec 11.
Artículo en Inglés | MEDLINE | ID: mdl-38139839

RESUMEN

Among the currently available commercial eye drops with cyclosporine A (Cs) there is a lack of long-acting dosage forms and products with a concentration of the drug substance higher than 0.1%, although Cs is widely used in ophthalmology. The aim of the research was to conduct the microscopic and biopharmaceutical evaluation of two formulations, an emulsion (EM) and a self-emulsifying oil (SEO), both with 0.5% of Cs, proposed for use in eye drops, and the comparison of both. SEO eye drops with Cs or any other drug substance are currently not available as marketed products, and the highest concentration of Cs in the ocular emulsion is only 0.1%. The microscopic evaluation of the emulsion and the SEO after emulsification with water was carried out using a high-resolution digital microscopy. The properties of both preparations were compared using the high dynamic range function or optical shadow effect mode. Images in the 3D composition mode were also recorded. The in vivo study of the Cs formulations was performed on male albino rabbits. The eye tolerance of the preparations was assessed using the ocular irritation test, which is a modified Draize test. Placebo carriers (without the drug substance) were also subjected to irritation testing. The concentration of Cs in the tissues (cornea and conjunctiva) and fluids (tear fluid and aqueous humor) of the rabbit eye was determined after multiple instillations of Cs-EM or Cs-SEO. The tested preparations were compared using the digital microscopy technique, which highlights the features of the formulations and eliminates the risk of unnoticeable properties that are difficult to observe in classical optical microscopy. Both tested Cs-loaded formulations are classified as practically non-irritating. There were also no significant differences when testing the placebo carriers. After a topical administration, Cs was widely distributed in all tissues (e.g., in cornea 1.3 ng/mg and 1.0 ng/mg) and fluids of the eye (e.g., in tear fluid 11.6 µg/mL and 4.3 µg/mL), after the administration of Cs-SEO and Cs-EM, respectively. The obtained results allow us to recognize both tested formulations, the emulsion and the self-emulsifying oil with 0.5% Cs content, as carriers safe for ophthalmic use and effective in delivering the drug substance to the structures of the eye.

19.
Cancers (Basel) ; 14(5)2022 Feb 25.
Artículo en Inglés | MEDLINE | ID: mdl-35267519

RESUMEN

The incidence of bladder cancer (BCa) has remained high for many years. Nevertheless, its pathomechanism has not yet been fully understood and is still being studied. Therefore, multiplatform untargeted urinary metabolomics analysis has been performed in order to study differences in the metabolic profiles of urine samples collected at three time points: before transurethral resection of bladder tumor (TURBT), the day after the procedure and two weeks after TURBT. Collected samples were analyzed with the use of high-performance liquid chromatography hyphenated with time-of-flight mass spectrometry detection (HPLC-TOF/MS) and gas chromatography coupled with triple quadrupole mass spectrometry detection (GC-QqQ/MS, in a scan mode). Levels of metabolites selected in our previous study were assessed in order to confirm their potential to differentiate the healthy and diseased samples, regardless of the risk factors and individual characteristics. Hippuric acid, pentanedioic acid and uridine confirmed their potential for sample differentiation. Based on the results of statistical analysis for the paired samples (comparison of metabolic profiles of samples collected before TURBT and two weeks after), a set of metabolites belonging to nucleotide metabolism and methylation processes was also selected. Longitudinal studies proved to be useful for the evaluation of metabolic changes in bladder cancer.

20.
Materials (Basel) ; 15(15)2022 Jul 27.
Artículo en Inglés | MEDLINE | ID: mdl-35955145

RESUMEN

Diatoms represent, in terms of species number, one of the largest groups of microalgae that have the ability to synthesize phenomenal mineral composites characterized by complex hierarchical structures. Their shells, called frustules, create intricately ornamented structures, reminiscent of the most sophisticated, natural mosaics. Ordinated pore systems perforate siliceous walls of the frustules with diameters ranging from nano to micro-scale, forming openwork three-dimensional silica structures. The use of these features is one of the main challenges in developing new technological solutions. In this study we assess the ability of selected diatom species (Pseudostaurosira trainorii) for metabolic insertion of soluble titanium from the culture medium into the structure of amorphous silica cell walls by its cultivation in laboratory conditions. The study is aimed at obtaining new and strengthening the already existing optical properties of diatomaceous biosilica. The physicochemical properties of the obtained materials have been studied using a series of instrumental methods.

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