Assessment of thimerosal effects in sublethal concentrations on zebrafish (Danio rerio) embryos exploring NMR-based metabolomics profile.

Thimerosal, a preservative commonly used in the pharmaceutical and cosmetic industry, has raised concerns regarding its potentially toxic effects as an organic mercury compound. Within this context, using an NMR-based metabolomics profile and chemometric analysis, zebrafish embryos were used as an in vivo model to study the effects of thimerosal in metabolic profiles after exposure to sublethal concentrations of the mercury compound. The thimerosal concentrations of 40 and 80 nM were employed, corresponding to 40% and 80% of the LC50, respectively, for zebrafish embryos. The most significant alterations in the metabolic profile included changes in carbohydrates, amino acids, nucleotides, trimethylamine-N-oxide, ethanolamine, betaine, and ethanol. Furthermore, thimerosal exposure affects various metabolic pathways, impairing the nervous system, disrupting protein metabolism, and potentially causing oxidative damage. Therefore, adopting a metabolomics approach in this investigation provided insights into the potentially implicated metabolic pathways contributing to the deleterious effects of thimerosal in biological systems.

NMR-based metabolomics applied to ecotoxicology with zebrafish (Danio rerio) as a prominent model for metabolic profiling and biomarker discovery: Overviewing the most recent approaches.

Metabolomics is an innovative approach used in the medical, toxicological, and biological sciences. As an interdisciplinary topic, metabolomics and its relation with the environment and toxicological research are extensive. The use of substances, such as drugs and pesticides, contributes to the continuous releasing of xenobiotics into the environment, harming organisms and their habitats. In this context, fish are important bioindicators of the environmental condition and have often been used as model species. Among them, zebrafish (Danio rerio) presents itself as a versatile and straightforward option due to its unique attributes for research. Zebrafish proves to be a valuable model for toxicity assays and also for metabolomics profiling by analytical tools. Thus, NMR-based metabolomics associated with statistical analysis can reasonably assist researchers in critical factors related to discovering and validating biomarkers through accurate diagnosis. Therefore, this review aimed to report the studies that applied zebrafish as a model for (eco)toxicological assays and essentially utilized NMR-based metabolomics analysis to assess the biochemical profile and thus suggest the potential biological marker.

Exosome-Enriched Plasma Analysis as a Tool for the Early Detection of Hypertensive Gestations.

Hypertensive disorders of pregnancy are closely associated with prematurity, stillbirth, and maternal morbidity and mortality. The onset of hypertensive disorders of pregnancy (HDP) is generally noticed after the 20th week of gestation, limiting earlier intervention. The placenta is directly responsible for modulating local and systemic physiology by communicating using mechanisms such as the release of extracellular vesicles, especially exosomes. In this study, we postulated that an analysis of exosome-enriched maternal plasma could provide a more focused and applicable approach for diagnosing HDP earlier in pregnancy. Therefore, the peripheral blood plasma of 24 pregnant women (11 controls, 13 HDP) was collected between 20th and 24th gestational weeks and centrifuged for exosome enrichment. Exosome-enriched plasma samples were analyzed by Raman spectroscopy and by proton nuclear magnetic resonance metabolomics (1H NMR). Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to analyze the Raman data, from the spectral region of 600-1,800 cm-1, to determine its potential to discriminate between groups. Using principal component analysis, we were able to differentiate the two groups, with 89% of all variances found in the first three principal components. In patients with HDP, most significant differences in Raman bands intensity were found for sphingomyelin, acetyl CoA, methionine, DNA, RNA, phenylalanine, tryptophan, carotenoids, tyrosine, arginine, leucine, amide I and III, and phospholipids. The 1H NMR analysis showed reduced levels of D-glucose, L-proline, L-tyrosine, glycine, and anserine in HDP, while levels of 2-hydroxyvalerate, polyunsaturated fatty acids, and very-low-density lipoprotein (VLDL) were increased. 1H NMR results were able to assign an unknown sample to either the control or HDP groups at a precision of 88.3% using orthogonal partial least squares discriminant analysis and 87% using logistic regression analysis. Our results suggested that an analysis of exosome-enriched plasma could provide an initial assessment of placental function at the maternal-fetal interface and aid HDP diagnosis, prognosis, and treatment, as well as to detect novel, early biomarkers for HDP.

USE OF METABOLOMICS TO THE DIAGNOSIS OF INFLAMMATORY BOWEL DISEASE.

BACKGROUND: Crohn's disease and ulcerative colitis are the primary inflammatory bowel diseases (IBD), and its pathogenesis is related to genetic and environmental factors. Currently, the diagnosis of IBD results in a multidisciplinary approach with significant disadvantages, such as its invasive nature, time spent, and the fact that 10% of patients remain without diagnostic classification. However, new methodologies of analysis have emerged that allowed the expansion of knowledge about IBD, as the metabolomics, the study of metabolites. The presence and prevalence of such metabolites may prove to be useful as biomarkers in the diagnosis of IBD. OBJECTIVE: Analyze fecal samples for metabolic analysis in the diagnosis of inflammatory bowel diseases (IBD), providing differentiation between Crohn's disease and ulcerative colitis. METHODS: This is an observational study with 21 patients diagnosed with IBD (ulcerative colitis 11 and Crohn's disease 10) and 15 healthy controls, all with the consent and clarification. The fecal extracts of all patients are submitted to a high-resolution Nuclear Magnetic Resonance Hydrogen (1H-NMR) spectroscopy combined with multivariate and univariate pattern recognition techniques. Through the metabolomics of fecal extracts, gives us a characterization of employing a noninvasive approach. RESULTS: We identify some metabolites, such as lactate, succinate, alanine, and tyrosine, in the Crohn's disease fecal samples, and leucine, alanine, and tyrosine in the ulcerative colitis fecal samples. All the amino acids presented positive covariance for disease correlation. CONCLUSION: The results showed different metabolic profiles between IBD patients and healthy volunteers based on 1H-NMR analysis of fecal extracts. Moreover, the approach discriminated patients with Crohn's disease and ulcerative colitis. The metabolomics analysis is promising as a novel diagnostic technique for further IBD recognition and surveillance. New studies are necessary to validate these findings.

Uso da metabolômica no diagnóstico de doenças inflamatórias intestinais / Use of metabolomics to the diagnosis of inflammatory bowel disease

ABSTRACT BACKGROUND: Crohn's disease and ulcerative colitis are the primary inflammatory bowel diseases (IBD), and its pathogenesis is related to genetic and environmental factors. Currently, the diagnosis of IBD results in a multidisciplinary approach with significant disadvantages, such as its invasive nature, time spent, and the fact that 10% of patients remain without diagnostic classification. However, new methodologies of analysis have emerged that allowed the expansion of knowledge about IBD, as the metabolomics, the study of metabolites. The presence and prevalence of such metabolites may prove to be useful as biomarkers in the diagnosis of IBD. OBJECTIVE: Analyze fecal samples for metabolic analysis in the diagnosis of inflammatory bowel diseases (IBD), providing differentiation between Crohn's disease and ulcerative colitis. METHODS: This is an observational study with 21 patients diagnosed with IBD (ulcerative colitis 11 and Crohn's disease 10) and 15 healthy controls, all with the consent and clarification. The fecal extracts of all patients are submitted to a high-resolution Nuclear Magnetic Resonance Hydrogen (1H-NMR) spectroscopy combined with multivariate and univariate pattern recognition techniques. Through the metabolomics of fecal extracts, gives us a characterization of employing a noninvasive approach. RESULTS: We identify some metabolites, such as lactate, succinate, alanine, and tyrosine, in the Crohn's disease fecal samples, and leucine, alanine, and tyrosine in the ulcerative colitis fecal samples. All the amino acids presented positive covariance for disease correlation. CONCLUSION: The results showed different metabolic profiles between IBD patients and healthy volunteers based on 1H-NMR analysis of fecal extracts. Moreover, the approach discriminated patients with Crohn's disease and ulcerative colitis. The metabolomics analysis is promising as a novel diagnostic technique for further IBD recognition and surveillance. New studies are necessary to validate these findings.

RESUMO CONTEXTO: A doença de Crohn e retocolite ulcerativa são as principais doenças inflamatórias intestinais (DII), e sua patogênese está relacionada a fatores genéticos e ambientais. Atualmente, o diagnóstico de DII resulta em uma abordagem multidisciplinar e apresenta desvantagens significativas, como sua natureza invasiva, tempo gasto e o fato de 10% dos pacientes permanecerem sem classificação diagnóstica. No entanto, surgiram novas metodologias de análise que permitiram ampliar o conhecimento sobre a DII, como a metabolômica, o estudo dos metabólitos. A presença e a prevalência desses metabólitos podem ser úteis como biomarcadores no diagnóstico da DII. OBJETIVO: Avaliar as amostras fecais por análise metabolômica no diagnóstico de DII, diferenciando os perfis metabólicos entre doença de Crohn e retocolite ulcerativa. MÉTODOS: Estudo observacional com 36 indivíduos (doença de Crohn 11, retocolite ulcerativa 10 e 15 controles saudáveis), todos com consentimento esclarecido. Os extratos fecais de todos os pacientes são submetidos a uma espectroscopia de alta resolução por ressonância magnética nuclear de hidrogênio (1H-RMN) combinada com técnicas de reconhecimento de padrões multivariados e univariados. Por meio da metabolômica utilizando extratos fecais, foi possível obter uma caracterização adequada das doenças inflamatórias intestinais através de uma abordagem não invasiva. RESULTADOS: Foi possível identificar os seguintes metabólitos nos pacientes com doen­ça de Crohn: lactato, succinato, alanina e tirosina e, no grupo retocolite ulcerativa encontrou-se leucina, alanina e tirosina. Todos os aminoácidos apresentaram covariância positiva para a doença. CONCLUSÃO: Os resultados demonstraram diferentes perfis metabólicos entre pacientes com DII e voluntários saudáveis, com base na análise por 1H-RMN dos extratos fecais. Além disso, pacientes com doença de Crohn e retocolite ulcerativa também podem ser discriminados usando essa abordagem. A análise metabolômica é promissora como uma nova técnica não invasiva de diagnóstico para melhor reconhecimento das DII. Novos estudos são necessários para validar esses achados.