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1.
Clin Oral Investig ; 29(2): 102, 2025 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-39890717

RESUMO

OBJECTIVE: Saliva is a rich-bodily fluid with recognized clinical diagnosis roles. This research aimed to investigate the salivary proteomic signatures for MIH in children with distinct degrees of severity. MATERIALS AND METHODS: In this cross-sectional study, 50 schoolers (8-13 years) were equally assigned into the following groups: G1 (Control group-Healthy first permanent molars), G2 (Mild MIH with white/creamy opacity and free of caries), G3 (Mild MIH with yellow/brown opacity and free of caries), G4 (Severe MIH with white/creamy, yellow/brown opacities including post-eruptive fracture and free of caries), G5 (Severe MIH with white/creamy, yellow/brown opacities, post-eruptive fracture, and caries). Unstimulated saliva samples were collected and later explored using mass spectrometry analysis. RESULTS: In total, 6,471 proteins were found, 5,073 exclusively from MIH children, and 778 overlapping among the different degrees of the disturb. The biological pathways displayed distinct patterns among the groups, which differed according to the MIH degrees. Gene-Ontology differences might not be verified regarding the biological processes and cellular components. Conversely, concerning molecular function, alterations among groups were evident, with proteins that would contribute to MIH in children with the severe condition (i.e., calcium ion binding, microtubule binding, platelet-derived growth factor binding). CONCLUSION: The results of this study support important salivary proteomics changes in MIH children according to distinct degrees of severity, reinforcing the interplay between the clinical characteristics and changes in the salivary proteome. CLINICAL RELEVANCE: Changes occurring in the salivary proteomics of children with distinct degrees of severity of Molar Incisor Hypomineralization (MIH) might be promising biomarkers and valuable information in clinical care, helping professionals make better clinical decisions and helping patients to understand their disturbance.


Assuntos
Hipoplasia do Esmalte Dentário , Proteômica , Saliva , Humanos , Saliva/química , Saliva/metabolismo , Feminino , Criança , Adolescente , Masculino , Estudos Transversais , Hipoplasia do Esmalte Dentário/metabolismo , Biomarcadores , Espectrometria de Massas , Dente Molar , Índice de Gravidade de Doença , Hipomineralização Molar
2.
Parasit Vectors ; 18(1): 27, 2025 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-39863914

RESUMO

BACKGROUND: Nippostrongylus brasiliensis-a nematode of rodents-is commonly used as a model to study the immunobiology of parasitic nematodes. It is a member of the Strongylida-a large order of socioeconomically important parasitic nematodes of animals. Lipids are known to play essential roles in nematode biology, influencing cellular membranes, energy storage and/or signalling. METHODS: The present investigation provides a comprehensive, untargeted lipidomic analysis of four developmental stages/sexes (i.e. egg, L3, adult female and adult male stages) of N. brasiliensis utilising liquid chromatography coupled to mass spectrometry. RESULTS: We identified 464 lipid species representing 18 lipid classes and revealed distinct stage-specific changes in lipid composition throughout nematode development. Triacylglycerols (TGs) dominated the lipid profile in the egg stage, suggesting a key role for them in energy storage at this early developmental stage. As N. brasiliensis develops, there was a conspicuous transition toward membrane-associated lipids, including glycerophospholipids (e.g. PE and PC) and ether-linked lipids, particularly in adult stages, indicating a shift toward host adaptation and membrane stabilisation. CONCLUSIONS: We provide a comprehensive insight into the lipid composition and abundance of key free-living and parasitic stages of N. brasiliensis. This study provides lipidomic resources to underpin the detailed exploration of lipid biology in this model parasitic nematode.


Assuntos
Lipidômica , Lipídeos , Nippostrongylus , Animais , Feminino , Masculino , Lipídeos/análise , Metabolismo dos Lipídeos , Infecções por Strongylida/parasitologia , Infecções por Strongylida/metabolismo , Estágios do Ciclo de Vida , Cromatografia Líquida , Triglicerídeos/metabolismo , Espectrometria de Massas
3.
J Chromatogr A ; 1743: 465715, 2025 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-39864224

RESUMO

Although proteins in snake venoms have been extensively studied and characterized, low-mass molecules remain relatively unexplored, mainly due to their low abundance, secondary role in envenomation, and some analytical technique limitations. However, these small molecules can provide new important data related to venom toxins' molecular structure, functions, and evolutionary relationships. This research aimed to characterize molecules below 10 kDa in the venoms of snakes from the Viperidae families (Bothrops, Agkistrodon, and Bitis) and compare two chromatographic approaches: reverse-phase chromatography (RP), a classic technique, and hydrophilic interaction liquid chromatography (HILIC), an alternative technique, both coupled with high-resolution mass spectrometry (HRMS). The results showed that the separation of the HILIC column provided a more efficient evenly distributed ion profile than RP, contributing to a 25.6% increase in the sequences identified. Homologous sequences for Bradykinin-potentiating peptides (BPPs) and fragments of major venom proteins, possibly cryptids, were found. In addition, BPP 13a, peptides rich in histidine and glycine (pHpG), and spacer sequences were identified in all snakes analyzed, especially with HILIC separation, suggesting that these sequences may be conserved within Viperidae. These findings indicate that the use of the HILIC column, compared to RP, is a promising approach for characterizing peptides in snake venom obtained by the ultrafiltration process. It contributes to the study of these still poorly understood molecules and is also a good option for studying other complex protein/peptide mixtures.


Assuntos
Interações Hidrofóbicas e Hidrofílicas , Espectrometria de Massas , Peptídeos , Viperidae , Animais , Espectrometria de Massas/métodos , Peptídeos/análise , Peptídeos/química , Cromatografia Líquida/métodos , Cromatografia de Fase Reversa/métodos , Venenos de Víboras/química , Sequência de Aminoácidos
4.
Clin Oral Investig ; 29(1): 77, 2025 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-39847108

RESUMO

OBJECTIVE: This study aimed to compare the salivary protein profile in individuals with Type 2 Diabetes Mellitus (DM2) and periodontitis and their respective controls. METHODS: Eighty participants were included in the study. The four groups were formed by individuals with DM2 and periodontitis (DM2 + P, n = 20), DM2 without periodontitis (DM2, n = 20), periodontitis without DM2 (P, n = 20) and individuals without periodontitis and without DM2 (H, n = 20). Periodontal clinical examinations were performed and unstimulated saliva was collected. Proteomic analysis was performed by shotgun mass spectrometry. The results were obtained by searching the Homo sapiens database of the UniProt catalog. RESULTS: A total of 220 proteins were identified in saliva samples. In the comparison between DM2 + P and DM2 groups, 27 proteins were up-regulated [e.g. S100-A8 was 6 times up-regulated (humoral immune response pathway)]. The DM2 + P and P groups had 26 up-regulated proteins [e.g. Immunoglobulin lambda constant 7 more than 2 times up-regulated (complement activation pathway)]. The non-DM2 groups (P and H) presented 22 up-regulated proteins [e.g. Glyceraldehyde-3-phosphate dehydrogenase more than 2 times up-regulated (Peptidyl-cysteine S-nitrosylation pathway)]. The groups without periodontitis (DM2 and H) showed 23 were up-regulated proteins [e.g. Hemoglobin subunit alpha that was more than 10 times up-regulated (cellular oxidant detoxification pathway)]. CONCLUSION: The presence of DM2 and periodontitis significantly impacts the salivary proteome. Our proteomic analysis demonstrated that changes in the S100 family proteins (S100A8 and S100 A9) are highly related to the presence of DM2 and periodontitis. CLINICAL RELEVANCE: Diabetes Mellitus (DM) and periodontitis are highly prevalent chronic diseases that present a wide variety of signs and symptoms. They present a bidirectional relationship, where patients with DM have a higher prevalence and severity of periodontitis, and patients with periodontitis have a higher prevalence of DM, worse glycemic control, and more diabetic complications. Diagnosing periodontitis requires specific clinical examinations, which require a highly trained operator. In this study, we used high throughput proteomics in order to evaluate non-invasive biomarkers for periodontitis in type 2 DM subjects. The results can contribute to earlier, more accurate, and less costly diagnosis of periodontitis in diabetic subjects, enabling better diabetes control.


Assuntos
Diabetes Mellitus Tipo 2 , Periodontite , Proteômica , Saliva , Humanos , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/complicações , Feminino , Saliva/metabolismo , Saliva/química , Masculino , Periodontite/metabolismo , Pessoa de Meia-Idade , Estudos de Casos e Controles , Proteínas e Peptídeos Salivares/metabolismo , Proteínas e Peptídeos Salivares/análise , Espectrometria de Massas , Adulto , Biomarcadores/metabolismo , Biomarcadores/análise
5.
J Agric Food Chem ; 72(50): 27876-27883, 2024 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-39630117

RESUMO

Cafestol is an ent-kaurene skeleton diterpene that is present in coffee beans and brews. Although several biological activities have been described in the literature for cafestol, such as hypercholesterolemic, anti-inflammatory, anticerous, and antidiabetic effects, its metabolism within the human body remains poorly understood. Therefore, this study aimed to quantify cafestol in boiled coffee brew, assess its bioaccessibility using a static in vitro digestion model, and investigate the metabolites formed during the digestion process using liquid chromatography coupled to high-resolution mass spectrometry. Cafestol content in the boiled coffee brew ranged from 127.47 to 132.65 mg L-1. The bioaccessibility of cafestol from boiled coffee brew using the in vitro digestion model was 93.65%; additionally, in the intestinal phase, cafestol was mainly found in its alcohol form. Additionally, a novel carboxylic acid derivative metabolite from cafestol with m/z 331.1909 [M + H]+ formed in the oral digestion phase is proposed. This metabolite was also detected in other digestion phases. Thus, this is the first article to investigate the metabolism of cafestol during digestion using an in vitro digestion model. The results indicate that cafestol is bioaccessible, is available to absorption, in its alcohol form, and suffers an oxidation reaction during the oral phase of digestion.


Assuntos
Coffea , Café , Digestão , Espectrometria de Massas , Modelos Biológicos , Humanos , Coffea/química , Coffea/metabolismo , Café/química , Café/metabolismo , Espectrometria de Massas/métodos , Sementes/química , Sementes/metabolismo , Diterpenos/metabolismo , Diterpenos/química , Diterpenos/análise , Diterpenos/farmacocinética , Disponibilidade Biológica , Cromatografia Líquida de Alta Pressão , Culinária
6.
Dalton Trans ; 53(47): 18880-18889, 2024 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-39508517

RESUMO

For years, cancer has been the second cause of death worldwide, preceded by cardiovascular diseases only. The number of research groups focusing on the discovery of new drugs to treat cancer is growing and the aim is to look for more effective compounds that cause less severe side effects and do not suffer from therapeutic resistance. The metal complexes cisplatin and carboplatin are widely used in the chemotherapeutic treatment of various types of cancer, including triple-negative breast cancer (TNBC). Both compounds are essential in modern chemotherapy and continue to be the subject of research to optimize their therapeutic properties and minimize adverse effects. Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) allows obtaining both quantitative data and information on the spatial distribution of elements in biological tissues and populations of single cells. In this work, the content of Pt and its distribution in TNBC MDA-MB-231 cells were determined via LA-ICP-MS analysis after incubation with different Pt-containing drugs. The quantitative analysis of single cells and the elemental maps revealing the distribution of Pt over and within the cells provide an enhanced insight into the potential mechanism of action of these Pt-containing drugs and their efficacy against TNBC.


Assuntos
Antineoplásicos , Terapia a Laser , Espectrometria de Massas , Humanos , Antineoplásicos/farmacologia , Antineoplásicos/química , Linhagem Celular Tumoral , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/patologia , Feminino , Carboplatina/farmacologia , Carboplatina/química , Cisplatino/farmacologia , Platina/química , Análise de Célula Única , Neoplasias da Mama/patologia , Neoplasias da Mama/tratamento farmacológico
7.
Sci Rep ; 14(1): 29570, 2024 11 28.
Artigo em Inglês | MEDLINE | ID: mdl-39609505

RESUMO

Mass spectrometry (MS)-based metabolomics analysis is a powerful tool, but it comes with its own set of challenges. The MS workflow involves multiple steps before its interpretation in what is denominate data mining. Data mining consists of a two-step process. First, the MS data is ordered, arranged, and presented for filtering before being analyzed. Second, the filtered and reduced data are analyzed using statistics to remove further variability. This holds true particularly for MS-based untargeted metabolomics studies, which focused on understanding fold changes in metabolic networks. Since the task of filtering and identifying changes from a large dataset is challenging, automated techniques for mining untargeted MS-based metabolomic data are needed. The traditional statistics-based approach tends to overfilter raw data, which may result in the removal of relevant data and lead to the identification of fewer metabolomic changes. This limitation of the traditional approach underscores the need for a new method. In this work, we present a novel deep learning approach using node embeddings (powered by GNNs), edge embeddings, and anomaly detection algorithm to analyze the data generated by mass spectrometry (MS)-based metabolomics called GEMNA (Graph Embedding-based Metabolomics Network Analysis), for example for an untargeted volatile study on Mentos candy, the data clusters produced by GEMNA were better than the ones used traditional tools, i.e., GEMNA has [Formula: see text], vs. the traditional approach has [Formula: see text].


Assuntos
Algoritmos , Mineração de Dados , Espectrometria de Massas , Metabolômica , Metabolômica/métodos , Espectrometria de Massas/métodos , Mineração de Dados/métodos , Humanos , Aprendizado Profundo , Redes e Vias Metabólicas , Metaboloma
8.
Int J Mol Sci ; 25(22)2024 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-39595969

RESUMO

Omics approaches were extensively applied during the coronavirus disease 2019 (COVID-19) pandemic to understand the disease, identify biomarkers with diagnostic and prognostic value, and discover new molecular targets for medications. COVID-19 continues to challenge the healthcare system as the virus mutates, becoming more transmissible or adept at evading the immune system, causing resurgent epidemic waves over the last few years. In this study, we used saliva from volunteers who were negative and positive for COVID-19 when Omicron and its variants became dominant. We applied a direct solid-phase extraction approach followed by non-target metabolomics analysis to identify potential salivary signatures of hospital-recruited volunteers to establish a model for COVID-19 screening. Our model, which aimed to differentiate COVID-19-positive individuals from controls in a hospital setting, was based on 39 compounds and achieved high sensitivity (85%/100%), specificity (82%/84%), and accuracy (84%/92%) in training and validation sets, respectively. The salivary diagnostic signatures were mainly composed of amino acids and lipids and were related to a heightened innate immune antiviral response and an attenuated inflammatory profile. The higher abundance of thyrotropin-releasing hormone in the COVID-19 positive group highlighted the endocrine imbalance in low-severity disease, as first reported here, underscoring the need for further studies in this area.


Assuntos
Biomarcadores , COVID-19 , Metabolômica , SARS-CoV-2 , Saliva , Humanos , COVID-19/metabolismo , COVID-19/diagnóstico , COVID-19/virologia , Saliva/metabolismo , Saliva/virologia , Metabolômica/métodos , Masculino , Feminino , SARS-CoV-2/isolamento & purificação , Adulto , Pessoa de Meia-Idade , Biomarcadores/metabolismo , Espectrometria de Massas/métodos , Idoso
9.
Nat Methods ; 21(12): 2327-2335, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-39482464

RESUMO

Advancing data analysis tools for proteome-wide cross-linking mass spectrometry (XL-MS) requires ground-truth standards that mimic biological complexity. Here we develop well-controlled XL-MS standards comprising hundreds of recombinant proteins that are systematically mixed for cross-linking. We use one standard dataset to guide the development of Scout, a search engine for XL-MS with MS-cleavable cross-linkers. Using other, independent standard datasets and published datasets, we benchmark the performance of Scout and existing XL-MS software. We find that Scout offers an excellent combination of speed, sensitivity and false discovery rate control. The results illustrate how our large recombinant standard can support the development of XL-MS analysis tools and evaluation of XL-MS results.


Assuntos
Espectrometria de Massas , Proteoma , Proteômica , Proteínas Recombinantes , Ferramenta de Busca , Proteínas Recombinantes/química , Espectrometria de Massas/métodos , Proteômica/métodos , Reagentes de Ligações Cruzadas/química , Humanos , Software
10.
J Food Sci ; 89(11): 7452-7463, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39390639

RESUMO

Ginger, which is the rhizome of Zingiber officinale Roscoe, is widely distributed and consumed. The taste and aroma of ginger differ depending on its geographical origin. To distinguish the origin of ginger, ginger extracts from Korea, Peru, and China were analyzed using ultra-performance liquid chromatography (UPLC) coupled to quadrupole time-of-flight mass spectrometry and nuclear magnetic resonance spectroscopy for metabolomics. Korean ginger contained more 10-gingerol, and Peruvian ginger contained more 6-gingerol and 8-gingerol. Several amino acids negatively correlated with gingerols, suggesting that amino acids are related to the biosynthesis of gingerols. Sugars, which are the main energy source, positively correlated with gingerols. Organic acids and gingerols were also positively correlated, indicating that both organic acids and gingerols are used for adaptation to the environment surrounding the root. We confirmed the features of the primary and secondary metabolites by verifying the correlation between metabolites and differences in metabolites according to ginger origin. We additionally optimized a simultaneous UPLC analytical method of marker compounds for the simple and rapid quality control of ginger. This method exhibits excellent linearity, sensitivity, and reproducibility. Using metabolomics, differences in origin were observed, and a low-end equipment analysis method for quality control can be used in the ginger industry.


Assuntos
Catecóis , Álcoois Graxos , Metabolômica , Fenóis , Extratos Vegetais , Zingiber officinale , Zingiber officinale/química , Metabolômica/métodos , Catecóis/metabolismo , Catecóis/análise , Álcoois Graxos/metabolismo , Álcoois Graxos/análise , Cromatografia Líquida de Alta Pressão/métodos , Fenóis/análise , Fenóis/metabolismo , Extratos Vegetais/química , China , Peru , Rizoma/química , República da Coreia , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas/métodos , Reprodutibilidade dos Testes
11.
Sci Data ; 11(1): 1105, 2024 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-39384817

RESUMO

This data descriptor presents a curated dataset for pathogen detection and identification (Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans) directly from whole-blood samples. The dataset was created using differential cell lysis combined with rapid extraction, digestion, and mass spectrometry-based proteomics. Our method offers a rapid diagnostic alternative to traditional culture, enabling timely disease management, such as sepsis. Highlighting our dataset's uniqueness, it features a three-tier structure: Spectral Libraries of Pathogens for identifying peptide peaks for putative biomarkers; Spiked pathogen in blood MS data for biomarker panel optimization through varied concentration samples; and Parallel Reaction Monitoring (PRM) data from sepsis patients for validating our biomarker panel, achieving 83.3% sensitivity within seven hours without microbial enrichment culture. This dataset serves as a comprehensive reference for bioinformatic tool development and biomarker panel proposals, advancing microbial detection, antimicrobial resistance, and epidemiological studies.


Assuntos
Biomarcadores , Candida albicans , Proteômica , Pseudomonas aeruginosa , Sepse , Staphylococcus aureus , Humanos , Proteômica/métodos , Biomarcadores/sangue , Sepse/sangue , Sepse/diagnóstico , Sepse/microbiologia , Espectrometria de Massas
12.
Braz J Biol ; 84: e283740, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39230082

RESUMO

More catechins are found in green tea than in any other type of tea, with its predominant production taking place in Asian nations. Consumption of green tea has been strongly correlated with a reduced risk of many diseases. This study introduces a new, efficient, and reliable method for extracting total catechins using ultra-high-performance liquid chromatography coupled with an ID-X-Orbitrap Mass spectrometer (UHPLC-IDX-Orbitrap-MS). The method was then applied to quantify the catechin content in green tea, yielding results comparable to previously published studies. Among the various sources of green tea analyzed, the lowest average catechin content was observed in Vietnam, Japan (2: Matcha), and Morocco, ranging between 346 and 322 mg/L. Conversely, the highest average catechin content (between 424 and 422 mg/L) was found in Sri Lanka and Japan (1: Sencha). For the remaining green tea extracts, the catechin levels ranged from 367 to 410 mg/L, exhibiting similar values. These findings demonstrate the high reproducibility of the proposed extraction procedure, with a relative standard deviation (RSD) error of less than 15% for the catechin standard. Additionally, the limit of detection for catechins was determined to be 1 ng mL-1. This study serves as a pilot investigation for extracting catechins from various green tea sources. Future research will focus on identifying all active compounds present. Furthermore, it is worth noting that this study aligns with the goals set forth in Saudi Vision 2030, which aims to diversify the country's economy and promote scientific advancements in various fields, including healthcare and agriculture.


Assuntos
Catequina , Espectrometria de Massas , Folhas de Planta , Chá , Catequina/análise , Catequina/isolamento & purificação , Cromatografia Líquida de Alta Pressão/métodos , Chá/química , Espectrometria de Massas/métodos , Reprodutibilidade dos Testes , Folhas de Planta/química , Extratos Vegetais/química , Extratos Vegetais/análise
13.
Mikrochim Acta ; 191(10): 620, 2024 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-39320493

RESUMO

Magnetic particle spray mass spectrometry (MPS-MS), an innovative ambient ionization technique proposed by our research group, was employed to determine beta-blockers in human plasma samples. A dispersive solid phase extraction of atenolol, metoprolol, labetalol, propranolol, nadolol, and pindolol was carried out using magnetic molecularly imprinted polymer (M-MIP) particles that were attached to the tip of a metal probe, which was placed in the mass spectrometer inlet. A solvent (1% formic acid in methanol) was dispensed on the particles, and the Taylor cone was formed around them (in high voltage). The analytes were desorbed/ionized and determined by a triple quadrupole mass spectrometer. M-MIP was synthesized with oxprenolol as a pseudo-template, demonstrating good selectivity to beta-blockers compared with no-analog molecules, with an adsorption process occurring in monolayers, according to isotherm studies. Kinetic experiments indicated chemisorption as the predominant M-MIP/analyte interaction. The analytical curves were linear (R2 > 0.98), and the limit of quantification was 3 µg L-1 for all the analytes. Limits of detection ranged from 0.64 to 2.41 µg L-1. Precisions (relative standard deviation) and accuracies (relative error) ranged from 3.95 to 21.20% and -17.05 to 18.93%, respectively. MPS-MS proved to be a simple, sensitive, and advantageous technique compared with conventional approaches. The analyses were fast, requiring no chromatographic separation and without ionic suppression. The method is aligned with green chemistry principles, requiring minimal sample, solvent, and sorbent amounts. MPS-MS successfully integrates sample preparation and ambient ionization mass spectrometry and holds great potential for application with other sorbents, samples, and analytes.


Assuntos
Antagonistas Adrenérgicos beta , Antagonistas Adrenérgicos beta/sangue , Antagonistas Adrenérgicos beta/química , Humanos , Limite de Detecção , Polímeros Molecularmente Impressos/química , Extração em Fase Sólida/métodos , Espectrometria de Massas/métodos , Adsorção
14.
Arq Bras Oftalmol ; 88(1): e20230037, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39109736

RESUMO

PURPOSE: To characterize the extracellular vesicle protein cargo in the aqueous humor and plasma of patients with ocular toxoplasmosis. METHODS: Aqueous humor and plasma were collected from six patients with active ocular toxoplasmosis and six patients with cataract. Extracellular vesicles were isolated, and western blotting and mass spectrometry were performed for protein analysis. RESULTS: All plasma samples from patients with ocular toxoplasmosis and cataract were positive for the tetraspanins CD63 and TSG101. However, the aqueous humor from patients with ocular toxoplasmosis was positive only for CD63. Sixty-seven new unreported proteins were identified in the aqueous humor and plasma of patients with the ocular toxoplasmosis and cataract. Of the 67 proteins, 10 and 7 were found only in the cataract and ocular toxoplasmosis groups, respectively. In general, these proteins were involved in immune system activation and retina homeostasis and were related to infections and retina-associated diseases. CONCLUSION: The distinct protein signatures between ocular toxoplasmosis and cataract may be helpful in the differential diagnosis of ocular toxoplasmosis. However, more studies are needed to better understand the role of these proteins in the pathogenesis of ocular toxoplasmosis.


Assuntos
Humor Aquoso , Western Blotting , Catarata , Vesículas Extracelulares , Toxoplasmose Ocular , Humanos , Humor Aquoso/metabolismo , Humor Aquoso/química , Humor Aquoso/parasitologia , Vesículas Extracelulares/metabolismo , Masculino , Feminino , Catarata/metabolismo , Pessoa de Meia-Idade , Adulto , Tetraspanina 30/análise , Tetraspanina 30/metabolismo , Espectrometria de Massas , Idoso , Proteínas de Ligação a DNA , Fatores de Transcrição , Complexos Endossomais de Distribuição Requeridos para Transporte
15.
Anal Bioanal Chem ; 416(24): 5233-5253, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39158631

RESUMO

Liquid chromatography-mass spectrometry (LC-MS) has emerged as a powerful analytical technique for analyzing complex biological samples. Among various chromatographic stationary phases, porous graphitic carbon (PGC) columns have attracted significant attention due to their unique properties-such as the ability to separate both polar and non-polar compounds and their stability through all pH ranges and to high temperatures-besides the compatibility with LC-MS. This review discusses the applicability of PGC for SPE and separation in LC-MS-based analyses of human biological samples, highlighting the diverse applications of PGC-LC-MS in analyzing endogenous metabolites, pharmaceuticals, and biomarkers, such as glycans, proteins, oligosaccharides, sugar phosphates, and nucleotides. Additionally, the fundamental principles underlying PGC column chemistry and its advantages, challenges, and advances in method development are explored. This comprehensive review aims to provide researchers and practitioners with a valuable resource for understanding the capabilities and limitations of PGC columns in LC-MS-based analysis of human biological samples, thereby facilitating advancements in analytical methodologies and biomedical research.


Assuntos
Grafite , Espectrometria de Massas , Humanos , Grafite/química , Cromatografia Líquida/métodos , Porosidade , Espectrometria de Massas/métodos , Extração em Fase Sólida/métodos , Biomarcadores/análise , Proteínas/análise , Polissacarídeos/análise , Espectrometria de Massa com Cromatografia Líquida
16.
Clinics (Sao Paulo) ; 79: 100459, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39098147

RESUMO

OBJECTIVE: Sjögren's Syndrome (SS) is a chronic inflammatory autoimmune exocrinopathy, and although, the role of metabolism in the autoimmune responses has been discussed in diseases such as lupus erythematosus, rheumatoid arthritis, psoriasis and scleroderma. There is a lack of information regarding the metabolic implications of SS. Considering that the disease affects primarily salivary glands; the aim of this study is to evaluate the metabolic changes in the salivary glands' microenvironment using a targeted metabolomics approach. METHODS: The saliva from 10 patients diagnosed with SS by the American-European consensus and 10 healthy volunteers was analyzed in an Ultra-high Performance Liquid Chromatograph Coupled Mass Spectrometry (UPLC-MS). RESULTS: The results showed an increased concentration in SS of metabolites involved in oxidative stress such as lactate, alanine and malate, and amino acids involved in the growth and proliferation of T-cells, such as arginine, leucine valine and isoleucine. CONCLUSIONS: These results revealed that is possible to differentiate the metabolic profile of SS and healthy individuals using a small amount of saliva, which in its turn may reflect the cellular changes observed in the microenvironments of damaged salivary glands from these patients.


Assuntos
Metabolômica , Saliva , Síndrome de Sjogren , Humanos , Síndrome de Sjogren/metabolismo , Saliva/química , Saliva/metabolismo , Metabolômica/métodos , Feminino , Pessoa de Meia-Idade , Estudos de Casos e Controles , Adulto , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas , Masculino , Estresse Oxidativo/fisiologia , Aminoácidos/análise , Aminoácidos/metabolismo , Idoso
17.
An Acad Bras Cienc ; 96(suppl 2): e20231140, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39194007

RESUMO

The aim of this study is to analyze the waterborne metal levels in four lakes (one endorheic and three exorheic) of Harmony Point, Nelson Island, Antarctica. Water samples were analyzed by using a quadrupole type inductively coupled plasma mass spectrometer and an inductively coupled plasma optical emission spectrometer. The levels of As, Cu, Mn, Mo, and V were significantly lower and those of Cr, Mg, Na, and Sr were significantly higher in the endorheic lake than in the other lakes. Most water samples presented levels of Ag, Be, Cd, Pb, Se, Tl, and U below the limit of quantification, while for Ba, Co, and Ni around half of the samples were below this limit. The waterborne metal levels were not significantly different between the exorheic lakes. Waterborne metal levels in the freshwater lakes from Harmony Point did not show any clear relationship with their levels in the soil of the region or with bird guano, and overall, their levels indicate an environment without anthropogenic influence. Apparently, the Na levels are influenced by salt spray from the ocean, as they are related to the distance of the lakes from the ocean.


Assuntos
Monitoramento Ambiental , Lagos , Poluentes Químicos da Água , Regiões Antárticas , Lagos/química , Lagos/análise , Monitoramento Ambiental/métodos , Poluentes Químicos da Água/análise , Metais/análise , Metais Pesados/análise , Espectrometria de Massas
18.
J Mass Spectrom ; 59(7): e5063, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38953332

RESUMO

An unprecedented and direct PS-MS (paper spray ionization mass spectrometry) method was proposed for the detection of native peptides, that is, glutathiones (GSHs), homoglutathiones (hGSHs), and phytochelatins (PCs), in basil (Ocimum basilicum L.) roots before and after cadmium exposure. The roots were submitted to cold maceration followed by sonication with formic acid as the extractor solvent for sample preparation. PS-MS was used to analyze such extracts in the positive mode, and the results allowed for the detection of several GSHs, hGSHs, and PCs. Some of these PCs were not distinguished in the control samples, that is, basil roots not exposed to cadmium. Other PCs were noticed in both types of roots, uncontaminated and cadmium-contaminated, but the intensities were higher in the former samples. Moreover, long-time exposure to cadmium stimulated the formation of some of these PCs and their cadmium complexes. The results, therefore, provided some crucial insights into the defense mechanism of plants against an external stress condition due to exposure to a toxic heavy metal. The present study represents a promising alternative to investigate other crucial physiological processes in plants submitted to assorted stress conditions.


Assuntos
Cádmio , Ocimum basilicum , Fitoquelatinas , Raízes de Plantas , Fitoquelatinas/química , Fitoquelatinas/metabolismo , Raízes de Plantas/química , Cádmio/análise , Ocimum basilicum/química , Espectrometria de Massas/métodos , Glutationa/análise , Glutationa/metabolismo , Glutationa/química
19.
PeerJ ; 12: e17757, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39076775

RESUMO

Soldiers of the Mexican Army with obesity were subjected to an intense 60-day weight-loss course consisting of a controlled diet, daily physical training, and psychological sessions. The nutritional treatment followed the European Society of Cardiology (ESC) recommendations, incorporating elements of the traditional milpa diet in the nutritional intervention. The total energy intake was reduced by 200 kcal every 20 days, starting with 1,800 kcal and ending with 1,400 kcal daily. On average, the participants reduced their body weight by 18 kg. We employed an innovative approach to monitor the progress of the twelve soldiers who completed the entire program. We compared the untargeted metabolomics profiles of their urine samples, taken before and after the course. The data obtained through liquid chromatography and high-resolution mass spectrometry (LC-MS) provided insightful results. Classification models perfectly separated the profiles pre and post-course, indicating a significant reprogramming of the participants' metabolism. The changes were observed in the C1-, vitamin, amino acid, and energy metabolism pathways, primarily affecting the liver, biliary system, and mitochondria. This study not only demonstrates the potential of rapid weight loss and metabolic pathway modification but also introduces a non-invasive method for monitoring the metabolic state of individuals through urine mass spectrometry data.


Assuntos
Militares , Obesidade , Redução de Peso , Humanos , Masculino , Obesidade/metabolismo , Obesidade/dietoterapia , Obesidade/terapia , Redução de Peso/fisiologia , Adulto , Metabolômica , Adulto Jovem , Metabolismo Energético/fisiologia , Espectrometria de Massas , Dieta Redutora , Programas de Redução de Peso/métodos , Reprogramação Metabólica
20.
Arch Toxicol ; 98(10): 3491-3502, 2024 10.
Artigo em Inglês | MEDLINE | ID: mdl-38951190

RESUMO

Snake venoms are complex mixtures majorly composed of proteins with well-studied biological effects. However, the exploration of non-protein components, especially lipids, remains limited despite their potential for discovering bioactive molecules. This study compares three liquid-liquid lipid extraction methods for both chemical and biological analyses of Bothrops moojeni snake venom. The methods evaluated include the Bligh and Dyer method (methanol, chloroform, water), considered standard; the Acunha method, a modification of the Bligh and Dyer protocol; and the Matyash method (MTBE/methanol/water), featuring an organic phase less dense than the aqueous phase. Lipidomic analysis using liquid chromatography with high-resolution mass spectrometry (LC-HRMS) system revealed comparable values of lipid constituents' peak intensity across different extraction methods. Our results show that all methods effectively extracted a similar quantity of lipid species, yielding approximately 17-18 subclasses per method. However, the Matyash and Acunha methods exhibited notably higher proportions of biologically active lipids compared to the Bligh and Dyer method, particularly in extracting lipid species crucial for cellular structure and function, such as sphingomyelins and phosphatidylinositol-phosphate. In conclusion, when selecting a lipid extraction method, it is essential to consider the study's objectives. For a biological approach, it is crucial to evaluate not only the total quantity of extracted lipids but also their quality and biological activity. The Matyash and Acunha methods show promise in this regard, potentially offering a superior option for extracting biologically active lipids compared to the Bligh and Dyer method.


Assuntos
Bothrops , Lipidômica , Lipídeos , Animais , Lipídeos/análise , Lipídeos/química , Lipidômica/métodos , Venenos de Crotalídeos/química , Cromatografia Líquida/métodos , Extração Líquido-Líquido/métodos , Espectrometria de Massas
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