RESUMO
Medium-chain acyl-CoA dehydrogenase deficiency (MCADD) is the most prevalent mitochondrial fatty acid ß-oxidation disorder. In this study, we assessed the variability of the lipid profile in MCADD by analysing plasma samples obtained from 25 children with metabolically controlled MCADD (following a normal diet with frequent feeding and under l-carnitine supplementation) and 21 paediatric control subjects (CT). Gas chromatography-mass spectrometry was employed for the analysis of esterified fatty acids, while high-resolution C18-liquid chromatography-mass spectrometry was used to analyse lipid species. We identified a total of 251 lipid species belonging to 15 distinct lipid classes. Principal component analysis revealed a clear distinction between the MCADD and CT groups. Univariate analysis demonstrated that 126 lipid species exhibited significant differences between the two groups. The lipid species that displayed the most pronounced variations included triacylglycerols and phosphatidylcholines containing saturated and monounsaturated fatty acids, specifically C14:0 and C16:0, which were found to be more abundant in MCADD. The observed changes in the plasma lipidome of children with non-decompensated MCADD suggest an underlying alteration in lipid metabolism. Therefore, longitudinal monitoring and further in-depth investigations are warranted to better understand whether such alterations are specific to MCADD children and their potential long-term impacts.
Assuntos
Acil-CoA Desidrogenase , Erros Inatos do Metabolismo Lipídico , Lipidômica , Fosfolipídeos , Triglicerídeos , Humanos , Erros Inatos do Metabolismo Lipídico/sangue , Lipidômica/métodos , Criança , Masculino , Feminino , Triglicerídeos/sangue , Fosfolipídeos/sangue , Pré-Escolar , Acil-CoA Desidrogenase/deficiência , Lactente , Adolescente , Metabolismo dos Lipídeos , Estudos de Casos e Controles , Ácidos Graxos/sangue , Ácidos Graxos/metabolismo , Cromatografia Gasosa-Espectrometria de Massas/métodos , Carnitina/sangueRESUMO
Extracellular vesicles (EVs) (exossomes, microvesicles and apoptotic bodies) have been well acknowledged as mediators of intercellular communications in prokaryotes and eukaryotes. Lipids are essential molecular components of EVs but at the moment the knowledge about the lipid composition and the function of lipids in EVs is limited and as for now none lipidomic studies in Giardia EVs was described. Therefore, the focus of the current study was to conduct, for the first time, the characterization of the polar lipidome, namely phospholipid and sphingolipid profiles of G. lamblia trophozoites, microvesicles (MVs) and exosomes, using C18-Liquid Chromatography-Mass Spectrometry (C18-LC-MS) and Tandem Mass Spectrometry (MS/MS). A total of 162 lipid species were identified and semi-quantified, in the trophozoites, or in the MVs and exosomes belonging to 8 lipid classes, including the phospholipid classes phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI), cardiolipins (CL), the sphingolipid classes sphingomyelin (SM) and ceramides (Cer), and cholesterol (ST), and 3 lipid subclasses that include lyso PC (LPC), lyso PE (LPE) and lyso PG (LPG), but showing different abundances. This work also identified, for the first time, in G. lamblia trophozoites, the lipid classes CL, Cer and ST and subclasses of LPC, LPE and LPG. Univariate and multivariate analysis showed clear discrimination of lipid profiles between trophozoite, exosomes and MVs. The principal component analysis (PCA) plot of the lipidomics dataset showed clear discrimination between the three groups. Future studies focused on the composition and functional properties of Giardia EVs may prove crucial to understand the role of lipids in host-parasite communication, and to identify new targets that could be exploited to develop novel classes of drugs to treat giardiasis.
Assuntos
Vesículas Extracelulares , Gastrópodes , Giardia lamblia , Giardíase , Animais , Lipidômica , Espectrometria de Massas em Tandem , Giardia , Ceramidas , Lecitinas , Fosfolipídeos , Esfingolipídeos , CardiolipinasRESUMO
Lipidomics represent a valid complementary tool to the biochemical analysis of plasma in humans. However, in cetaceans, these tools have been unexplored. Here, we evaluated how the plasma lipid composition of Tursiops truncatus is modulated by developmental stage and sex, aiming at a potential use of lipidomics in integrated strategies to monitor cetacean health. We characterized the fatty acid profile and detected a total of 26 fatty acids in T. truncatus plasma. The most abundant fatty acids were palmitic acid (C16:0), stearic acid (C18:0) and oleic acid (C18:1n-9). Interestingly, there are consistent differences between the fatty acid profile of mature female and mature male specimens. Phospholipidome analysis identified 320 different lipid species belonging to phosphatidylcholine (PC, 105 lipid species), lysophosphatidylcholine (42), phosphatidylethanolamine (PE, 67), lysophosphatidylethanolamine (18), phosphatidylglycerol (14), lysophosphatidylglycerol (8), phosphatidylinositol (14), lysophosphatidylinositol (2), phosphatidylserine (3), sphingomyelin (45) and ceramides (2) classes. The statistical analysis of the phospholipidome showed that its composition allows discriminating mature animals between sexes and mature males from immature males. Notably, discrimination between sexes is mainly determined by the contents of PE plasmalogens and lysophospholipids (LPC and LPE), while the differences between mature and immature male animals were mainly determined by the levels of PC lipids. This is the first time that a correlation between developmental stage and sex and the lipid composition of the plasma has been established in cetaceans. Being able to discern between age and sex-related changes is an encouraging step towards using these tools to also detect differences related to disease/dysfunction processes.
Assuntos
Golfinho Nariz-de-Garrafa , Humanos , Animais , Masculino , Feminino , Ácidos Graxos , Lisofosfatidilcolinas , CeramidasRESUMO
In recent years, several studies have demonstrated that polyunsaturated fatty acids have strong immunomodulatory properties, altering several functions of macrophages. In the present work, we sought to provide a multi-omic approach combining the analysis of the lipidome, the proteome, and the metabolome of RAW 264.7 macrophages supplemented with phospholipids containing omega-3 (PC 18:0/22:6; ω3-PC) or omega-6 (PC 18:0/20:4; ω6-PC) fatty acids, alone and in the presence of lipopolysaccharide (LPS). Supplementation of macrophages with ω3 and ω6 phospholipids plus LPS produced a significant reprogramming of the proteome of macrophages and amplified the immune response; it also promoted the expression of anti-inflammatory proteins (e.g., pleckstrin). Supplementation with the ω3-PC and ω6-PC induced significant changes in the lipidome, with a marked increase in lipid species linked to the inflammatory response, attributed to several pro-inflammatory signalling pathways (e.g., LPCs) but also to the pro-resolving effect of inflammation (e.g., PIs). Finally, the metabolomic analysis demonstrated that supplementation with ω3-PC and ω6-PC induced the expression of several metabolites with a pronounced inflammatory and anti-inflammatory effect (e.g., succinate). Overall, our data show that supplementation of macrophages with ω3-PC and ω6-PC effectively modulates the lipidome, proteome, and metabolome of these immune cells, affecting several metabolic pathways involved in the immune response that are triggered by inflammation.
Assuntos
Ácidos Graxos Ômega-3/metabolismo , Ácidos Graxos Ômega-6/metabolismo , Fatores Imunológicos/metabolismo , Lipídeos/fisiologia , Macrófagos/metabolismo , Fosfolipídeos/metabolismo , Proteínas/metabolismo , Animais , Imunidade/fisiologia , Inflamação/metabolismo , Lipidômica/métodos , Metaboloma/fisiologia , Camundongos , Proteoma/metabolismo , Células RAW 264.7 , Transdução de Sinais/fisiologiaRESUMO
OBJECTIVE: This study evaluated the effect of different glucose concentration on biofilm formation of Candida albicans and Candida glabrata strains isolated from diabetic and non-diabetic individuals. METHODS: The study was divided into two stages: (I) selection and identification of 48 C. albicans and C. glabrata strains by polymerase chain reaction followed by restriction fragment length polymorphism analysis (PCR/RFLP); (II) evaluation of biofilm formation by means of viability rates (colony-forming units), biofilm dry matter (mg) and biofilm-covered areas (µm2). Statistical comparisons were performed through nonparametric analysis of longitudinal data in factorial experiments with pairwise comparisons using Friedman Conover's test (α = 0.05). RESULTS: All the Candida spp. had their identifications confirmed by PCR/RFLP. C. albicans biofilm of strains from diabetic individuals cultivated in different glucose concentration showed higher viability rates than strains from non-diabetic individuals. No difference was observed on viability of C. glabrata biofilm. Regarding biofilm dry matter, C. albicans biofilm of strains from diabetic individuals cultivated in different glucose concentration showed lower amount in weight than strains from non-diabetic individuals. In C. glabrata strains, this result was only observed in biofilms cultivated with no glucose supplementation. With regard to biofilm-covered areas, only glucose supplementation and non-diabetic condition showed a positive effect on C. albicans biofilm development, and no condition affected C. glabrata biofilm formation. CONCLUSION: The strain type (C. albicans and C. glabrata) isolated from diabetic and non-diabetic individuals influenced on biofilm formation, but glucose supplementation did not.
Assuntos
Candida albicans , Diabetes Mellitus , Antifúngicos , Biofilmes , Candida glabrata , Suplementos Nutricionais , Glucose , HumanosRESUMO
Noncommunicable diseases (NCD) and age-associated diseases (AAD) are some of the gravest health concerns worldwide, accounting for up to 70% of total deaths globally. NCD and AAD, such as diabetes, obesity, cardiovascular disease, and cancer, are associated with low-grade chronic inflammation and poor dietary habits. Modulation of the inflammatory status through dietary components is a very appellative approach to fight these diseases and is supported by increasing evidence of natural and dietary components with strong anti-inflammatory activities. The consumption of bioactive lipids has a positive impact on preventing chronic inflammation and consequently NCD and AAD. Thus, new sources of bioactive lipids have been sought out. Microalgae are rich sources of bioactive lipids such as omega-6 and -3 polyunsaturated fatty acids (PUFA) and polar lipids with associated anti-inflammatory activity. PUFAs are enzymatically and non-enzymatically catalyzed to oxylipins and have a significant role in anti and pro-resolving inflammatory responses. Therefore, a large and rapidly growing body of research has been conducted in vivo and in vitro, investigating the potential anti-inflammatory activities of microalgae lipids. This review sought to summarize and critically analyze recent evidence of the anti-inflammatory potential of microalgae lipids and their possible use to prevent or mitigate chronic inflammation.
Assuntos
Envelhecimento/efeitos dos fármacos , Misturas Complexas/uso terapêutico , Ácidos Graxos Ômega-3/uso terapêutico , Ácidos Graxos Ômega-6/uso terapêutico , Microalgas/química , Doenças não Transmissíveis/tratamento farmacológico , Misturas Complexas/química , Ácidos Graxos Ômega-3/química , Ácidos Graxos Ômega-6/química , Humanos , Inflamação/tratamento farmacológicoRESUMO
Grateloupia turuturu Yamada, 1941, is a red seaweed widely used for food in Japan and Korea which was recorded on the Atlantic Coast of Europe about twenty years ago. This seaweed presents eicosapentaenoic acid (EPA) and other polyunsaturated fatty acids (PUFAs) in its lipid fraction, a feature that sparked the interest on its potential applications. In seaweeds, PUFAs are mostly esterified to polar lipids, emerging as healthy phytochemicals. However, to date, these biomolecules are still unknown for G. turuturu. The present work aimed to identify the polar lipid profile of G. turuturu, using modern lipidomics approaches based on high performance liquid chromatography coupled to high resolution mass spectrometry (LC-MS) and gas chromatography coupled to mass spectrometry (GC-MS). The health benefits of polar lipids were identified by health lipid indices and the assessment of antioxidant and anti-inflammatory activities. The polar lipids profile identified from G. turuturu included 205 lipid species distributed over glycolipids, phospholipids, betaine lipids and phosphosphingolipids, which featured a high number of lipid species with EPA and PUFAs. The nutritional value of G. turuturu has been shown by its protein content, fatty acyl composition and health lipid indices, thus confirming G. turuturu as an alternative source of protein and lipids. Some of the lipid species assigned were associated to biological activity, as polar lipid extracts showed antioxidant activity evidenced by free radical scavenging potential for the 2,2'-azino-bis-3-ethyl benzothiazoline-6-sulfonic acid (ABTSâ+) radical (IC50 ca. 130.4 µg mL-1) and for the 2,2-diphenyl-1-picrylhydrazyl (DPPHâ) radical (IC25 ca. 129.1 µg mL-1) and anti-inflammatory activity by inhibition of the COX-2 enzyme (IC50 ca. 33 µg mL-1). Both antioxidant and anti-inflammatory activities were detected using a low concentration of extracts. This integrative approach contributes to increase the knowledge of G. turuturu as a species capable of providing nutrients and bioactive molecules with potential applications in the nutraceutical, pharmaceutical and cosmeceutical industries.
Assuntos
Antioxidantes/farmacologia , Extratos Vegetais/farmacologia , Alga Marinha , Compostos de Bifenilo , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Ácidos Graxos Insaturados/farmacologia , Humanos , Lipidômica , Espectrometria de Massas , Fosfolipídeos/farmacologia , Picratos , Relação Estrutura-AtividadeRESUMO
Phenylketonuria (PKU) is a disease of the catabolism of phenylalanine (Phe), caused by an impaired function of the enzyme phenylalanine hydroxylase. Therapeutics is based on the restriction of Phe intake, which mostly requires a modification of the diet. Dietary restrictions can lead to imbalances in specific nutrients, including lipids. In the present study, the plasma phospholipidome of PKU and healthy children (CT) was analyzed by hydrophilic interaction liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry. Using this approach, 187 lipid species belonging to nine different phospholipid classes and three ceramides were identified. Principal component analysis of the lipid species data set showed a distinction between PKU and CT groups. Univariate analysis revealed that 146 species of phospholipids were significantly different between both groups. Lipid species showing significant variation included phosphatidylcholines, containing polyunsaturated fatty acids (PUFA), which were more abundant in PKU. The high level of PUFA-containing lipid species in children with PKU may be related to a diet supplemented with PUFA. This study was the first report comparing the plasma polar lipidome of PKU and healthy children, highlighting that the phospholipidome of PKU children is significantly altered compared to CT. However, further studies with larger cohorts are needed to clarify whether these changes are specific to phenylketonuric children.
Assuntos
Fenilcetonúrias , Criança , Dieta , Suplementos Nutricionais , Ácidos Graxos Insaturados , Humanos , Fenilalanina , Fenilcetonúrias/diagnósticoRESUMO
UVB phototherapy is treatment for psoriasis, which increases phospholipid oxidative modifications in the cell membrane of the skin. Therefore, we carried out lipidomic analysis on the keratinocytes of healthy individuals and patients with psoriasis irradiated with UVB and treated with cannabidiol (CBD), phytocannabinoid with antioxidant and anti-inflammatory properties. Our results showed that, in psoriatic keratinocytes phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidylserine (PS), and ether-linked phosphoethanolamine (PEo), were downregulated, while SM (d41:2) was upregulated. These changes were accompanied by an increase in negative zeta potential, which indicates translocation of PS to the outer layer of the membrane. CBD treatment of psoriatic keratinocytes led to downregulation of PC, PS, and upregulation of certain PEo and an SM species, SM (d42:2), and the zeta potential. However, UVB irradiation of psoriatic keratinocytes resulted in upregulation of PC, PC plasmalogens (PCp), PEo, and a decrease in the negative zeta potential. The exposure of UVB-irradiated cells to CBD led to a decrease in the level of SM (d42:2). Our results suggest that CBD induces pro-apoptotic mechanisms in psoriatic keratinocytes while simultaneously improving the antioxidant properties and preventing the loss of transepidermal water of keratinocytes of patients irradiated with UVB. Thus, CBD has potential therapeutic value in the treatment of psoriasis.
Assuntos
Canabidiol/uso terapêutico , Queratinócitos/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Fosfolipídeos/metabolismo , Psoríase/tratamento farmacológico , Adulto , Canabidiol/farmacologia , Estudos de Casos e Controles , Feminino , Humanos , Queratinócitos/metabolismo , Queratinócitos/efeitos da radiação , Metabolismo dos Lipídeos/efeitos da radiação , Masculino , Pessoa de Meia-Idade , Cultura Primária de Células , Psoríase/metabolismo , Psoríase/radioterapia , Raios Ultravioleta , Terapia Ultravioleta , Adulto JovemRESUMO
In the last decades, the use of algae in biotechnology and food industries has experienced an exponential growth. Codium tomentosum is a green macroalgae with high biotechnological potential, due to its rich lipidome, although few studies have addressed it. This study aimed to investigate the seasonal changes in lipid and pigment profiles of C. tomentosum, as well as to screen its antioxidant activity, in order to evaluate its natural plasticity. Samples of C. tomentosum were collected in two different seasons, early-autumn (September/October) and spring (May), in the Portuguese coast (wild samples), and in a land-based integrated multitrophic aquaculture (IMTA) system (IMTA samples). Total lipid extracts were analysed by LC-MS, GC-MS, and HPLC, and antioxidant activity was screened through free radical scavenging potential against DPPH and 2,20-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals. Wild samples showed a high seasonal variability, modifying their lipidome and pigment profiles according to environmental shifts, while IMTA samples showed a relatively stable composition due to early-stage culturing in controlled conditions. The lipids that contributed the most to seasonal discrimination were glycolipids (monogalactosyl diacylglycerol - MGDG and digalactosyl diacylglycerol - DGDG) and the lyso forms of phospholipids and glycolipids. Lipid extracts showed antioxidant activity ranging from 61 ± 2 to 115 ± 35 µmol Trolox g-1 of lipid extract in DPPH assay and from 532 ± 73 to 927 ± 92 µmol Trolox g-1 of lipid extract in ABTS assay, with a more intense antioxidant activity in wild spring samples. This study revealed that wild specimens of C. tomentosum presented a higher plasticity to cope with seasonal environmental changes, adjusting their lipid, pigment, and bioactivity profiles, while IMTA samples, cultured under controlled conditions, displayed more stable lipidome and pigment compositions.
Assuntos
Clorófitas/química , Sequestradores de Radicais Livres/farmacologia , Lipídeos/farmacologia , Extratos Vegetais/farmacologia , Alga Marinha/química , Aquicultura , Benzotiazóis/antagonistas & inibidores , Biotecnologia/métodos , Compostos de Bifenilo/antagonistas & inibidores , Cromatografia Líquida de Alta Pressão , Sequestradores de Radicais Livres/isolamento & purificação , Lipídeos/isolamento & purificação , Espectrometria de Massas , Picratos/antagonistas & inibidores , Extratos Vegetais/isolamento & purificação , Estações do Ano , Ácidos Sulfônicos/antagonistas & inibidoresRESUMO
Palmaria palmata is an edible red macroalga widely used for human consumption and valued for its high protein value. Despite its low total lipid content, it is rich in eicosapentaenoic acid (EPA). This seaweed has been scarcely explored with regard to its lipid composition. The polar lipids of seaweeds are nowadays recognized as important phytochemicals contributing to their add value valorization and providing support for claims of potential health benefits. The present study aimed to disclose the polar lipid profile of P. palmata, farmed in an integrated multi-trophic aquaculture (IMTA) through modern lipidomic approaches using high-resolution LC-MS and MS/MS and to screen for the antioxidant properties of this red macroalga. A total of 143 molecular species of lipids were identified, belonging to several classes of polar lipids, such as glycolipids, phospholipids, and betaine lipids. It is noteworthy that the most abundant lipid species in each class were esterified with eicosapentaenoic acid (EPA), accounting for more than 50% of the lipid content. The polar lipid extract rich in EPA showed antioxidant activity with an inhibition concentration (IC) of IC30 = 171 ± 19.8 µg/mL for α,α-diphenyl-ß-picrylhydrazyl radical (DPPHâ) and IC50 = 26.2 ± 0.1 µg/mL for 2,20-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical cation (ABTSâ+). Overall, this study highlights that P. palmata farmed in an IMTA framework can be a sustainable source of beneficial lipids with antioxidant activity. Moreover, this red macroalga can be exploited for future applications as a source of lipids rich in EPA for food and feed, nutraceuticals, and cosmetics.
Assuntos
Ácido Eicosapentaenoico/análise , Sequestradores de Radicais Livres/farmacocinética , Extratos Vegetais/farmacologia , Rodófitas/química , Alga Marinha/química , Aquicultura , Cromatografia Líquida de Alta Pressão , Cosméticos , Suplementos Nutricionais , Sequestradores de Radicais Livres/análise , Sequestradores de Radicais Livres/química , Alimento Funcional , Concentração Inibidora 50 , Lipidômica , Extratos Vegetais/análise , Extratos Vegetais/química , Espectrometria de Massas em TandemRESUMO
Under roasting conditions, polysaccharides depolymerize and also are able to polymerize, forming new polymers through non-enzymatic transglycosylation reactions (TGRs). TGRs can also occur between carbohydrates and aglycones, such as the phenolic compounds present in daily consumed foods like coffee. In this study, glycosidically-linked phenolic compounds were quantified in coffee melanoidins, the polymeric nitrogenous brown-colored compounds formed during roasting, defined as end-products of Maillard reaction. One third of the phenolics present were in glycosidically-linked form. In addition, the roasting of solid-state mixtures mimicking coffee beans composition allowed the conclusion that proteins play a regulatory role in TGRs extension and, consequently, modulate melanoidins composition. Overall, the results obtained showed that TGRs are a main mechanism of phenolics incorporation in melanoidins and are inhibited by amino groups through Maillard reaction.
Assuntos
Coffea/química , Fenóis/química , Polímeros/química , Carboidratos/química , Café/química , Glicosilação , Temperatura Alta , Reação de MaillardRESUMO
Three mixtures containing different molar proportions of (ß1â4)-D-mannotriose and (α1â5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. The identification of hybrid oligosaccharides allowed us to infer the occurrence of nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures. Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting.