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1.
Food Chem ; 398: 133885, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-35973299

RESUMO

This work studied the difference in pulp breakdown between two cultivars of longan cv. 'Dongbi' and 'Fuyan' from the aspect of metabolisms of lipid and energy. The results reflected that, compared to 'Fuyan' longan, 'Dongbi' longan had higher levels of energy charge, U/S and IUFA, and higher amounts of USFA, PC, PI, ATP and ADP. Moreover, 'Dongbi' longan exhibited lower levels of SFA, PA, AMP and cell membrane permeability. Also, lower PLD, LOX and lipase activities, but higher ATPase activity, lower pulp breakdown index, and better pulp appearance were exhibited in 'Dongbi' longan. These data revealed that the mitigated pulp breakdown in 'Dongbi' longan was due to the comprehensive coordination of metabolisms in lipid and energy through maintaining a higher level of energy, a higher unsaturation degree of fatty acids, delaying the degradation of phospholipids, and better retaining the membrane structural integrity of microsome and entire cell.


Assuntos
Frutas , Sapindaceae , Adenosina Trifosfatases/metabolismo , Frutas/química , Fosfolipídeos/análise , Sapindaceae/metabolismo
2.
Food Chem ; 398: 133953, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-35998486

RESUMO

The aim of this study was to evaluate the influence of l-ascorbyl palmitate (LAP) as an additive to liposome formulations by self-assembling with soy lecithin to form hybrid liposomes, in order to enhance the physical stability and bioactivator-loaded retention ratio of the LAP incorporated liposomes (LAP-LP). The addition of LAP significantly increased its surface negative charge and strong hydrophobic interactions occurred between the hydrophobic tails of LAP and phospholipids resulting in more compactly ordered, rigid and hydrophobic phospholipid bilayers as indicated by surface tension, fluorescence probes and DSC. These changes enhanced the stability of hydrophobic polyphenol loaded LAP-LP during storage. Particularly, after four weeks storage at 37 °C for naringenin loaded liposomes, the retention ratio of pure liposome decreased dramatically to 12.5 %, while the LAP-LP remained above 74.5 %. This study opens up the potential for the LAP-LP to be developed as a food-grade multifunctional formulation for encapsulating and delivering bioactivators.


Assuntos
Lipossomos , Fosfolipídeos , Ácido Ascórbico/análogos & derivados , Estabilidade de Medicamentos , Interações Hidrofóbicas e Hidrofílicas , Lipossomos/química , Fosfolipídeos/química , Polifenóis
3.
Food Chem ; 399: 133977, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-35994857

RESUMO

In this work, ultra-performance liquid chromatography-electrospray ionization mass spectrometry (UPLC-ESI-MS) was applied to compare the lipidomics profile of chicken breast meat heated to various temperatures. A total of 445 lipids including phospholipids, diacylglycerols, triacylglycerols, glycoglycerolipids, and sphingolipids were detected in chicken muscle. Significant decreases in a list of characteristic unsaturated phospholipid species including phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol were identified after heating, suggesting the occurrence of lipid oxidation, while significant increases in characteristic lysophospholipid species were found with increasing heating temperature, which were mainly derived from hydrolysis of phospholipids. Moreover, ether phospholipid and neutral lipid species exhibited remarkable changes during the heating process, and monoalk(en)yl diacylglycerol was first identified in chicken meat. The findings could contribute to the improved understanding of key lipids and biochemical reactions engaged in the heating of meat products.


Assuntos
Galinhas , Lipidômica , Animais , Carne , Fosfolipídeos , Espectrometria de Massas por Ionização por Electrospray
4.
Food Chem ; 399: 133982, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36027811

RESUMO

Lipase-catalyzed transesterification of structured phospholipids (sPLs) is a hot topic, but the structural variation of the fatty acyl chains in intact phospholipids at the molecular level remains unclear to date. The present study explored the detailed characteristics of synthesized phospholipids through high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in precursor ion scan mode. The optimal conditions were in-depth inspected and determined for the reaction system, including phospholipase A1 as catalyst, 15% lipase loading, and 1% water content. The sPLs enriched with EPA/DHA were structurally and quantitatively characterized by focusing on the fragments of m/z 301.6 (eicosapentaenoic acid, EPA) and m/z 327.6 (docosahexaenoic acid, DHA), and the results were statistically analyzed using partial least squares discriminant analysis and clustered heatmap hierarchical clustering analysis. PC 38:6 (18:1/20:5), PC 38:7 (18:2/20:5), PC o-40:6 (o-18:0/22:6), and PE 40:8 (18:2/22:6) etc. were revealed as the main variables that were active in the reaction.


Assuntos
Ácidos Graxos Ômega-3 , Fosfolipídeos , Catálise , Cromatografia Líquida de Alta Pressão , Ácidos Docosa-Hexaenoicos/química , Ácido Eicosapentaenoico/química , Ácidos Graxos Ômega-3/química , Lipase/genética , Fosfolipídeos/química , Espectrometria de Massas em Tandem
5.
Int J Lab Hematol ; 44 Suppl 1: 89-100, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36074709

RESUMO

Platelet procoagulant mechanisms are emerging to be complex and important to achieving haemostasis. The mechanisms include the release of procoagulant molecules from platelet storage granules, and strong agonist-induced expression of procoagulant phospholipids on the outer platelet membrane for tenase and prothrombinase assembly. The release of dense granule polyphosphate is important to platelet procoagulant function as it promotes the activation of factors XII, XI and V, inhibits tissue factor pathway inhibitor and fibrinolysis, and strengthens fibrin clots. Platelet procoagulant function also involves the release of partially activated factor V from platelets. Scott syndrome has provided important insights on the mechanisms that regulate procoagulant phospholipids expression on the external platelet membrane, which require strong agonist stimulation that increase cystolic calcium levels, mitochondrial calcium uptake, the loss of flippase function and activation of the transmembrane scramblase protein anoctamin 6. There have been advances in the methods used to directly and indirectly assess platelet procoagulant function in health and disease. Assessments of thrombin generation with platelet rich plasma samples has provided new insights on how platelet procoagulant function is altered in inherited platelet disorders, and how platelets influence the bleeding phenotype of a number of severe coagulation factor deficiencies. Several therapies, including desmopressin and recombinant factor VIIa, improve thrombin generation by platelets. There is growing interest in targeting platelet procoagulant function for therapeutic benefit. This review highlights recent advances in our understanding of platelet-dependent procoagulant mechanisms in health and in bleeding disorders.


Assuntos
Transtornos da Coagulação Sanguínea , Transtornos Hemorrágicos , Plaquetas/metabolismo , Cálcio/metabolismo , Humanos , Fosfolipídeos/metabolismo , Ativação Plaquetária , Trombina/metabolismo
6.
Arch Microbiol ; 204(10): 607, 2022 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-36076091

RESUMO

A novel Streptomyces strain, designated TRM 75549T, was isolated from a sample of sand in Pimo reclamation area, Taklimakan desert, Xinjiang, North-West China. Phylogenetic analyses of the 16S rRNA gene sequences placed strain TRM75549T within the genus Streptomyces with the highest similarities to Streptomyces pilosus NBRC 12772T (98.7%). Nonetheless, average nucleotide identity (ANI) value and the digital DNA-DNA hybridization (dDDH) value between strain TRM75549T and S. pilosus NBRC 12772T were, respectively, 88.2% and 44.1%, and well below 95-96% and 70% cutoff point recommended for recognizing genomospecies, respectively. A multi-locus sequence analysis of five house-keeping genes (atpD, gyrB, recA, rpoB and trpB) and phylogenomic analysis also illustrated that strain TRM75549T should also be assigned to the genus Streptomyces. Strain TRM75549T contained MK-9 (H6) and MK-9 (H8) as predominant menaquinones. The diagnostic diamino acid of cell walls were identified as LL-diaminopimelic acid and meso-diaminopimelic. The whole-cell sugar pattern of strain TRM 75549T consisted of mannose and glucose. The major fatty acids (> 5%) were iso-C14:0, iso-C15:0, anteiso-C15:0, iso-C16:1H, iso-C16:0. The polar lipids were diphosphatidylglycerol, lysophosphatidylglycerol, phosphatidylethanolamine, phospholipids, phosphatidylglycerol, phosphatidylinositol, phosphatiylinositol mannosides and unidentified phospholipids. Strain TRM75549T could be differentiated from S. pilosus NBRC 12772T, based on physiological and biochemical characteristics. Thus, strain TRM75549T is representative of a novel species of the genus Streptomyces, for which the name Streptomyces pimonensis sp. nov. is proposed. The type strain is TRM75549T (= CCTCC AA 2020054T = LMG 32221T).


Assuntos
Streptomyces , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/análise , Fosfolipídeos/análise , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/química
7.
Drug Deliv ; 29(1): 2784-2795, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36047012

RESUMO

Levocetirizine hydrochloride (LVC) is an antihistaminic drug that is repurposed for the treatment of alopecia. This investigation is targeted for formulating LVC into cationic ceramide/phospholipid composite (CCPCs) for the management of alopecia. CCPCs were fabricated by ethanol-injection approach, through a central composite experiment. CCPCs were evaluated by inspecting their entrapment efficiency (EE%), polydispersity index (PDI), particle size (PS), and zeta potential (ZP). The optimum CCPCs were additionally studied by in-vitro, ex-vivo, in-silico, and in-vivo studies. The fabricated CCPCs had acceptable EE%, PS, PDI, and ZP values. The statistical optimization elected optimum CCPCs composed of 5 mg hyaluronic acid, 10 mg ceramide III, and 5 mg dimethyldidodecylammonium bromide employing phytantriol as a permeation enhancer. The optimum CCPCs had EE%, PS, PDI, and ZP of 88.36 ± 0.34%, 479.00 ± 50.34 nm, 0.377 ± 0.0035, and 20.20 ± 1.13 mV, respectively. The optimum CCPC maintained its stability for up to 90 days. It also viewed vesicles of tube shape via transmission electron microscope. The in-silico assessment resulted in better interaction and stability between LVC and vesicle components in water. The ex-vivo and in-vivo assessments showed satisfactory skin retention of LVC from optimum CCPCs. The histopathological assessment verified the safety of optimum CCPCs to be topically applied. Overall, the optimum CCPCs could be utilized as a potential system for the topical management of alopecia, with a prolonged period of activity, coupled with reduced LVC shortcomings.


Assuntos
Ceramidas , Fosfolipídeos , Alopecia/tratamento farmacológico , Cetirizina , Portadores de Fármacos , Reposicionamento de Medicamentos , Humanos , Tamanho da Partícula
8.
Sci Rep ; 12(1): 15127, 2022 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-36068249

RESUMO

Evidence is mounting that the nature of the lipid bound to the endothelial cell protein C receptor (EPCR) has an impact on its biological roles, as observed in anticoagulation and more recently, in autoimmune disease. Phosphatidylethanolamine and phosphatidylcholine species dominate the EPCR lipid cargo, yet, the extent of diversity in the EPCR-associated lipid repertoire is still unknown and remains to be uncovered. We undertook mass spectrometry analyses to decipher the EPCR lipidome, and identified species not yet described as EPCR ligands, such as phosphatidylinositols and phosphatidylserines. Remarkably, we found further, more structurally divergent lipids classes, represented by ceramides and sphingomyelins, both in less abundant quantities. In support of our mass spectrometry results and previous studies, high-resolution crystal structures of EPCR in three different space groups point to a prevalent diacyl phospholipid moiety in EPCR's pocket but a mobile and ambiguous lipid polar head group. In sum, these studies indicate that EPCR can associate with varied lipid classes, which might impact its properties in anticoagulation and the onset of autoimmune disease.


Assuntos
Doenças Autoimunes , Fosfolipídeos , Anticoagulantes , Fatores de Coagulação Sanguínea , Receptor de Proteína C Endotelial , Humanos , Fosfolipídeos/química , Receptores de Superfície Celular
9.
Artigo em Inglês | MEDLINE | ID: mdl-36067072

RESUMO

A novel Gram-stain-negative, aerobic, coccus-shaped bacteria, designated ZY201115T, was isolated from the nasal cavity of a sheep with respiratory disease in Yunnan Province, south-west China, and its taxonomic affiliation was studied by applying a polyphasic approach. The strain grew at 18-41 °C (optimum, 37 °C), at pH 6.0-9.0 (optimum, pH 8.0) and in 0.5-3.0% (w/v) NaCl (optimum, 1.0 % NaCl). Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain is affiliated to the genus Moraxella with highest similarity to Moraxella bovis ATCC 10900T (96.6 %). Phylogenomic analysis based on 811 single-copy genes also indicated that the strain represents a novel species in the genus Moraxella and formed a deep and separated clade with Moraxella caviae NCTC 10293T. The highest genomic orthologous average nucleotide identity and digital DNA-DNA hybridization values between the strain and the type strains in the genus Moraxella were 73.7% (M. caviae NCTC 10293T) and 25.3% (Moraxella osloensis CCUG 350T), respectively. The G+C content of the complete genome sequence was 42.1 mol%. The predominant fatty acids (>5 %) were C18:1 ω9c, C17:1 ω8c, C12:03OH and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c). The major polar lipids were phosphatidylglycerol, cardiolipin, monolysocardiolipin, phosphatidylethanolamine and hemibismonoacylglycerophosphate. The major respiratory quinone was CoQ-8. On the basis of the results of phylogenetic, phenotypic and chemotaxonomic characterizations, strain ZY201115T clearly represents a novel species of the genus Moraxella, for which the name Moraxella nasovis sp. nov. is proposed. The type strain is ZY201115T (=CCTCC AB 2021473T=CCUG 75922T).


Assuntos
Ácidos Graxos , Cloreto de Sódio , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Moraxella/genética , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ovinos , Ubiquinona/química
10.
Arch Microbiol ; 204(9): 586, 2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-36048288

RESUMO

Members of the genus Marinomonas are known for their environmental adaptation and metabolically versatility, with abundant proteins associated with antifreeze, osmotic pressure resistance, carbohydrase and multiple secondary metabolites. Comparative genomic analysis focusing on secondary metabolites and orthologue proteins was conducted with 30 reference genome sequences in the genus Marinomonas. In this study, a Gram-stain-negative, rod-shaped, non-flagellated and strictly aerobic bacterium, designated as strain E8T, was isolated from the red algae (Gelidium amansii) in the coastal of Weihai, China. Optimal growth of the strain E8T was observed at temperatures 25-30 °C, pH 6.5-8.0 and 1-3% (w/v) NaCl. The DNA G + C content was 42.8 mol%. The predominant isoprenoid quinone was Q-8 and the major fatty acids were C16:0, summed feature 3 and summed feature 8. The major polar lipids were phosphatidylglycerol (PG) and phosphatidylethanolamine (PE). Based on data obtained from this polyphasic taxonomic study, strain E8T should be considered as a novel species of the genus Marinomonas, for which the name Marinomonas algarum is proposed. The type strain is E8T (= KCTC 92201T = MCCC 1K07070T).


Assuntos
Marinomonas , Rodófitas , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Ácidos Graxos/química , Genômica , Marinomonas/genética , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Rodófitas/genética , Rodófitas/microbiologia , Análise de Sequência de DNA , Ubiquinona/química
11.
Artigo em Inglês | MEDLINE | ID: mdl-36048630

RESUMO

Strain RHZ10T was isolated from an oak rhizosphere sampled in Reims, France, and characterized to assess its taxonomy. Based on 16S rRNA gene sequence similarity, strain RHZ10T was affiliated to the genus Streptomyces and the closest species were Streptomyces anulatus NRRL B-2000T and Streptomyces pratensis ch24T. Average nucleotide identity and digital DNA-DNA hybridization values were 77.3-92.4 % and 23.0-50.9 %, respectively, when compared to the type strains of fully sequenced related species having a 16S rRNA gene sequence similarity over 98 %. These data suggested that strain RHZ10T represented a novel species within the genus Streptomyces. The genome of RHZ10T was 8.0 Mbp long, had 7  894 predicted coding genes, and a G+C content of 71.7 mol%. Cultures of RHZ10T on ISP 2 medium mostly led to the production a green pigmentation of the core of its colonies in the vegetative mycelium, surrounded by white pigmentation of the aerial mycelium. The main fatty acids of RHZ10T were anteiso-C15 : 0, iso-C16 : 0, anteiso-C17 : 0 and C16 : 0. Polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, unidentified lipids, unidentified phospholipids, unidentified aminolipids and unidentified glycolipids. Its main quinones were MK-9(H6) (69.3 %), MK-9(H4) (17.3 %) and MK-9(H8) (17.0%). Phylogenetic, physiological and chemotaxonomic studies clearly supported that strain RHZ10T represents a novel species within the genus Streptomyces, for which the name Streptomyces durocortorensis sp. nov. is proposed and the type strain is RHZ10T (=DSM 112634T=LMG 32187T=CIP 111907T).


Assuntos
Quercus , Streptomyces , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos , Filogenia , RNA Ribossômico 16S/genética , Rizosfera , Análise de Sequência de DNA , Microbiologia do Solo
12.
J Chem Phys ; 157(8): 085103, 2022 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-36050012

RESUMO

Lipid peroxides result from a reaction between cis-unsaturated lipid chains and singlet oxygen molecules leading to the addition of a peroxide OOH side group next to the acyl-chain double bond. It is now established that HP-POPC (hydroperoxidized POPC) molecules form stable, thin, and laterally expanded bilayers. The difference in the structural organization arises from the hydrophilic character of the OOH side group that has a strong affinity with the water interface region, leading to significant reorganization of the bilayer. In this article, we describe a coarse-grained (CG) model of POPC and DOPC lipid peroxides within the framework of the Martini CG force-field (v2.2), derived from experimental data. We then discuss extensively the predicted structure and the influence of hydration and show how shifting the position of the unsaturated bonds along the chain changes the structure. Finally, we provide electron and neutron scattering length density profiles of the simulated bilayers.


Assuntos
Bicamadas Lipídicas , Fosfolipídeos , Interações Hidrofóbicas e Hidrofílicas , Bicamadas Lipídicas/química , Peróxidos Lipídicos , Fosfatidilcolinas/química , Fosfolipídeos/química , Água
13.
Arch Microbiol ; 204(9): 596, 2022 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-36056268

RESUMO

Strain DKSPLA3T, a novel Gram-negative, catalase-positive, oxidase-positive, non-spore-forming, aerobic, non-nitrogen-fixing, non-motile bacterium was isolated from Quercus variablis leaf, in Zunyi, Guizhou, China. Growth occurred at 4-37 °C (optimum 28 °C), pH 4.0-9.0 (optimum pH 7.0) and up to 4.0% (w/v) NaCl (optimum under 2.0%, w/v). Phylogeny based on 16S rRNA gene indicated that strain DKSPLA3T was a novel species in the genus Rhizobium, which was supported by average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values. The predominant fatty acids of strain DKSPLA3T were C16:0, C18:1 ω7c and/or C18:1 ω6c and C18:1 ω7c 11-methyl. The major respiratory quinone was Q-10. Major polar lipids were diphosphatidyl glycerol (DPG), phosphatidyl glycerol (PG), phosphatidylethanolamine (PE), phosphatidylmonomethylethanolamine (PME), phosphatidylcholine (PC), two unidentified phospholipids (PL) and nine unidentified lipids (L). The genomic G + C content was 64.47 mol%. Based on the phenotypic, phylogenetic and genotypic data, DKSPLA3T should be classified as a novel species in the genus Rhizobium, for which the name Rhizobium quercicola sp. nov. (KCTC 82843T = CFCC 16,707T) is proposed.


Assuntos
Quercus , Rhizobium , Técnicas de Tipagem Bacteriana , China , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Filogenia , Folhas de Planta , Quercus/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
14.
Ceska Slov Farm ; 71(3): 98-102, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36058638

RESUMO

The research was performed on 80 male rats of the Wistar line. Animals of two age groups were used: adults (10-12 months) and old (22-25 months). The obtained data show that the development of immobilization stress in adult and aged rats are accompanied by the formation of a characteristic complex of changes in the phospholipid composition of the myocardium. Intraperitoneal injection of the chokeberry extract (Aronia melanocarpa) at a dose of 0.2 g/kg 60 minutes before the immobilization has limited stress modulation of myocardial phospholipid composition in aged animals. Thus, the extract of Aronia melanocarpa increases the myocardial resistance to the injury effect of stress.


Assuntos
Photinia , Animais , Masculino , Miocárdio , Fosfolipídeos , Extratos Vegetais/farmacologia , Ratos , Ratos Wistar
15.
J Phys Chem B ; 126(36): 6936-6947, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-36066119

RESUMO

Hemocompatibility is one of the major criteria for the successful cardiovascular applicability of novel biomaterials. In this context, monolayers of certain biomolecules can be used to improve surface biocompatibility. To this end, biocoatings incorporating a phospholipid (1,2-dioleoyl-sn-glycero-3-phosphocholine, DOPC), an immunosuppressant (cyclosporine A, CsA), and an antioxidant material (lauryl gallate, LG) were fabricated by depositing Langmuir films onto gold or mica substrates using the Langmuir-Blodgett (LB) technique. These LB monolayers were thoroughly characterized by means of quartz crystal microbalance (QCM), atomic force microscopy (AFM), cyclic voltammetry (CV), and contact angle (CA) measurements. The obtained results indicate that the properties of these LB films are modulated by the monolayer composition. The presence of LG in the three-component systems (DOPC-CsA-LG) increases the molecular packing and the surface coverage of the substrate, which affects the wettability of the biocoating. From the different compositions studied here, we conclude that DOPC-CsA-LG monolayers with a DOPC/CsA ratio of 1:1 and LG molar fractions of 0.50 and 0.75 exhibit improved surface biocompatible characteristics. These results open up new perspectives on our knowledge and better understanding of phenomena at the biomaterial/host interface.


Assuntos
Antioxidantes , Fosfolipídeos , Imunossupressores , Microscopia de Força Atômica , Fosfolipídeos/química , Propriedades de Superfície , Molhabilidade
16.
Food Res Int ; 160: 111743, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36076425

RESUMO

Large yellow croaker roe phospholipids (LYCPLs) have various functions and high nutritional value, which are expected to be functional foods ingredients. The aim of this study is to investigate the oxidative stability of LYCPLs at different temperatures (100 °C, 140 °C and 180 °C) in terms of peroxide value (POV), thiobarbituric acid-reactive substances value (TBARS), acid value (AV) and fatty acids compositions of phospholipids. The volatile compounds were also identified and analyzed via headspace solid-phase microextraction-gas chromatography-mass spectroscopy (HS-SPME-GC-MS) and headspace gas chromatography-ion mobility spectrometry (HS-GC-IMS). The results showed that the highest POV was 192.92, 133.70 and 121.25 meq/kg at 100 °C, 140 °C and 180 °C within 10-20 min, respectively, then fell along with the increment of heating time, while the highest TBARS was 6.38, 7.66 and 8.19 mg/kg at 30 min, respectively. The AV was increased, but monounsaturated fatty acid and polyunsaturated fatty acid content were decreased with the increment of temperatures. A total of 94 volatile compounds were identified, and the content of alkanes, aldehydes, and heterocyclic compounds were reasonably increased, while the aromatic compound and amide compound content were decreased when LYCPLs heated at 180 °C. The present research improves the comprehensive understanding of oxidative stability of marine phospholipids.


Assuntos
Perciformes , Fosfolipídeos , Animais , Cromatografia Gasosa-Espectrometria de Massas/métodos , Estresse Oxidativo , Fosfolipídeos/química , Substâncias Reativas com Ácido Tiobarbitúrico
17.
J Pharm Biomed Anal ; 220: 115011, 2022 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-36054945

RESUMO

Lipid nanoparticles (LNPs) have shown great success as drug delivery systems, especially for mRNA vaccines, as those developed during the Covid-19 pandemics. Lipid analysis is critical to monitor the formulation process and control the quality of LNPs. The present study is focused on the development and validation of a high-performance liquid chromatography - diode array detector -evaporative light scattering detector (HPLC-DAD/ELSD) based method for the simultaneous quantification of 7 lipids, illustrating the main components of LNPs: ionizable lipids, the neutral co-lipid cholesterol, phospholipids, hydrophilic polymer-lipids for colloidal stability (e.g., a PEGylated lipid). In particular, this study focuses on two innovative synthetic lipids: a switchable cationic lipid (CSL3) which has demonstrated in vitro and in vivo siRNA transfection abilities, and the palmitic acid-grafted-poly(ethyloxazoline)5000 (PolyEtOx), used as an alternative polymer to address allergic reactions attributed to PEGylated lipids. The HPLC separation was achieved on a Poroshell C18 column at 50 °C using a step gradient of a mobile phase composed of water/methanol mixtures with 0.1% (v/v) trifluoroacetic acid (TFA). This method was validated following ICH Q2(R1) & (R2) guidelines in terms of linearity (R² ≥ 0.997), precision (relative standard deviation on peak areas < 5% for intermediate repeatability), accuracy (recoveries between 92.9% and 108.5%), and sensitivity. Indeed, low detection and quantitation limits were determined (between 0.02 and 0.04 µg and between 0.04 and 0.10 µg, respectively). Due to its high selectivity, this method allowed the analysis of lipid degradation products produced through degradation studies in basic, acidic, and oxidative conditions. Moreover, the method was successfully applied to the analysis of several liposome formulations at two key steps of the development process. Consequently, the reported HPLC method offers fast, versatile, selective and quantitative analysis of lipids, essential for development optimization, chemical characterization, and stability testing of LNP formulations.


Assuntos
COVID-19 , Nanopartículas , Colesterol , Cromatografia Líquida de Alta Pressão/métodos , Humanos , Lipossomos , Metanol , Nanopartículas/química , Ácido Palmítico , Fosfolipídeos , Polietilenoglicóis , Polímeros , RNA Interferente Pequeno , Ácido Trifluoracético , Água
18.
Cell Death Dis ; 13(9): 808, 2022 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-36130940

RESUMO

Triple-negative breast cancer (TNBC) is a heterogeneous subtype of breast cancer that displays highly aggressive with poor prognosis. Owing to the limited targets and drugs for TNBC clinical therapy, it is necessary to investigate the factors regulating cancer progression and develop novel therapies for cancer treatment. Ferroptosis, a nonapoptotic form of programmed cell death characterized by accumulation of iron-dependent peroxidation of phospholipids, is regulated by cellular metabolism, redox homeostasis, and various cancer-related signaling pathways. Recently, considerable progress has been made in demonstrating the critical role of lipid metabolism in regulating ferroptosis, indicating potential combinational therapeutic strategies for cancer treatment. In this study, by drug combination screen of lipid metabolism compounds with ferroptosis inducers in decreasing TNBC cell viability, we found potent synergy of the CB1 antagonist rimonabant with erastin/(1 S, 3 R)-RSL3 (RSL3) in inhibiting TNBC cell growth both in vitro and in vivo via promoting the levels of lipid peroxides, malondialdehyde (MDA), 4-hydroxynonenal (4-HNE) and cytosolic reactive oxygen species (ROS) production, enhancing intracellular glutathione (GSH) depletion and inducing G1 cell cycle arrest. We identified that inhibition of CB1 promoted the effect of erastin/RSL3 on inducing ferroptosis and enhanced their inhibitory effect on tumor growth. Using RNA-Seq, fatty acid analyses and functional assays, we found that CB1 regulated stearoyl-CoA desaturase 1 (SCD1)- and fatty acyl desaturase 2 (FADS2)-dependent fatty acid metabolism via phosphatidylinositol 3 kinase (PI3K)-AKT and mitogen-activated protein kinase (MAPK) signaling pathways to modulate ferroptosis sensitivity in TNBC cells. These data demonstrate that dual targeting of CB1 and ferroptosis could be a promising therapeutic strategy for TNBC.


Assuntos
Ferroptose , Neoplasias de Mama Triplo Negativas , Morte Celular , Ácidos Graxos/farmacologia , Glutationa/metabolismo , Humanos , Ferro/metabolismo , Metabolismo dos Lipídeos , Peróxidos Lipídicos , Malondialdeído , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosfolipídeos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Receptores de Canabinoides/metabolismo , Rimonabanto/farmacologia , Estearoil-CoA Dessaturase/metabolismo , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/metabolismo
19.
Anal Chem ; 94(36): 12292-12296, 2022 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-36048752

RESUMO

A key element of successful lipidomics analysis is a sufficient extraction of lipid molecules typically by two-phase systems such as chloroform-based Bligh and Dyer (B&D). However, numerous metabolomics and lipidomics studies today apply easy to use one-phase extractions. In this work, quantitative flow injection analysis high-resolution mass spectrometry was applied to benchmark the lipid recovery of popular one-phase extraction methods for human plasma samples. The following organic solvents were investigated: methanol (MeOH), ethanol (EtOH), 2-propanol (IPA), 1-butanol (BuOH), acetonitrile (ACN) and the solvent mixtures BuOH/MeOH (3:1) and MeOH/ACN (1:1). The recovery of polar lysophospholipids was sufficient for all tested solvents. However, nonpolar lipid classes such as triglycerides (TG) and cholesteryl esters (CE) revealed extraction efficiencies less than 5% due to precipitation in polar solvents EtOH, MeOH, MeOH/ACN, and ACN. Sample pellets also contained a substantial amount of phospholipids, for example, more than 75% of total phosphatidylcholine and sphingomyelin for ACN. The loss of lipids by precipitation was directly related to the polarity of solvents and lipid classes. Although, lipid recovery increased with the volume of organic solvent, recovery in polar MeOH remains incomplete also for less polar lipid classes such as ceramides. Addition of stable isotope-labeled internal standards prior to lipid extraction could compensate for insufficient lipid recovery for polar lipid classes including lysolipids and phospholipids but not for nonpolar CE and TG. In summary, application of one-phase extractions should be limited to polar lipid classes unless sufficient recovery/solubility of nonpolar lipids has been demonstrated. The presented data reveal that appropriate lipid extraction efficiency is fundamental to achieve accurate lipid quantification.


Assuntos
Benchmarking , Lipidômica , Humanos , Espectrometria de Massas/métodos , Metanol/química , Fosfolipídeos , Solventes/química , Triglicerídeos
20.
Cell Chem Biol ; 29(9): 1363-1365, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-36113403

RESUMO

ACSL4 is generally considered to be universally required for ferroptosis, a form of cell death induced by phospholipid peroxidation. In this issue of Cell Chemical Biology, Magtanong et al. (2022) report the surprising finding that ACSL4 is only required for ferroptosis in certain circumstances, raising interesting questions about the regulation of this cell death pathway.


Assuntos
Ferroptose , Morte Celular , Coenzima A Ligases/metabolismo , Fosfolipídeos
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