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1.
Ren Fail ; 45(1): 2171886, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36715439

RESUMO

OBJECTIVES: Subfornical organ (SFO) is vital in chronic kidney disease (CKD) progression caused by high salt levels. The current study investigated the effects of high salt on phosphoproteomic changes in SFO in CKD rats. METHODS: 5/6 nephrectomized rats were fed a normal-salt diet (0.4%) (NC group) or a high-salt diet (4%) (HC group) for three weeks, while sham-operated rats were fed a normal-salt diet (0.4%) (NS group). For phosphoproteomic analysis of SFO in different groups, TiO2 enrichment, isobaric tags for relative and absolute quantification (iTRAQ) labeling, and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used. RESULTS: There were 6808 distinct phosphopeptides found, which corresponded to 2661 phosphoproteins. NC group had 168 upregulated and 250 downregulated phosphopeptides compared to NS group. Comparison to NC group, HC group had 154 upregulated and 124 downregulated phosphopeptides. Growth associated protein 43 (GAP43) and heat shock protein 27 (Hsp27) were significantly upregulated phosphoproteins and may protect against high-salt damage. Differential phosphoproteins with tight functional connection were synapse proteins and microtubule-associated proteins, implying that high-salt diet disrupted brain's structure and function. Furthermore, differential phosphoproteins in HC/NC comparison group were annotated to participate in GABAergic synapse signaling pathway and aldosterone synthesis and secretion, which attenuated inhibitory neurotransmitter effects and increased sympathetic nerve activity (SNA). DISCUSSION: This large scale phosphoproteomic profiling of SFO sheds light on how salt aggravates CKD via the central nervous system.


Assuntos
Insuficiência Renal Crônica , Órgão Subfornical , Ratos , Animais , Ratos Sprague-Dawley , Cromatografia Líquida , Órgão Subfornical/fisiologia , Fosfopeptídeos/farmacologia , Espectrometria de Massas em Tandem , Cloreto de Sódio na Dieta/farmacologia , Fosfoproteínas/metabolismo , Fosfoproteínas/farmacologia
2.
J Hazard Mater ; 446: 130665, 2023 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-36592559

RESUMO

Exploring the behavior of pesticide residues in fruits is important for effectively applying pesticides and minimizing the risk of pesticide exposure to humans. However, most studies do not consider in situ visual analysis of residues and migration patterns in fresh fruit samples. We investigated the migration patterns of thiram, propamocarb, imidacloprid and pyraclostrobin in fresh bananas based on ambient mass spectrometry imaging, metabolome and transcriptome analysis. The systemic pesticides entered via lateral penetration and vertical migration over time, which began to internally migrate to the inner core after 6 h. The non-systemic pesticide thiram did not enter the interior of the bananas, and remained only in the peel. The transportation rate of the pesticides increased with the decrease of water-octanol partition coefficient and the relative molecular mass. Moreover, the pesticide migrated fast with the increase of banana ripeness. The pesticides significantly enhanced pyruvate kinase, NADP-dependent malic enzyme, and malate synthase activities in the banana peels through carbohydrate metabolism. The banana pulp was also protected against the external toxicity of pesticides by the ascorbate-glutathione cycle. These results can provide guidelines for the appropriate application of pesticides and their safety evaluation.


Assuntos
Musa , Resíduos de Praguicidas , Praguicidas , Humanos , Resíduos de Praguicidas/análise , Musa/química , Tiram/análise , Praguicidas/análise , Frutas/química , Metaboloma , Espectrometria de Massas em Tandem/métodos
3.
Sci Rep ; 13(1): 245, 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36604469

RESUMO

To identify biomarkers of hormonal contraceptive (HC) use in urine and saliva, we conducted a pilot study with 30 women initiating levonorgestrel (LNG) containing combined oral contraceptives (COCs) or depot medroxyprogesterone acetate (DMPA) (15/group). Based on established COC pharmacokinetics, we collected serum and urine samples before COC ingestion and during Days one and three of use, or before DMPA injection and on Days 21 and 60 post-injection. We used liquid chromatography-tandem mass spectrometry (LC-MS/MS) to measure serum/urine LNG and MPA. LNG was undetectable at baseline (specificity 100%); post ingestion, most urine samples had detectable LNG levels (sensitivity: 80% 6 h post Dose one, 93% 6 h post Dose three). We used a DetectX LNG immunoassay kit and showed 100% sensitivity measuring urine LNG. Urine MPA levels were undetectable in 14/15 women at baseline (specificity 91%); post-injection all urine samples had detectable MPA levels (sensitivity: 100% days 21 and 60). Results suggest urine sampling can be used to identify a biomarker of LNG and MPA use. Based on evidence from other steroidal hormonal studies showing changes affecting the transcriptome profile of saliva at 24 h, we used the same (COC, DMPA) timepoints to collect saliva. We performed transcriptome analysis and detected several differentially expressed genes in DMPA users' saliva on Days 21 and 60 compared to baseline; none among COC users. We plan further research of differential gene expression in saliva as a HC biomarker of DMPA use, and will explore longer periods of COC use and saliva collection times, and application of microRNA sequencing to support using saliva as a COC biomarker.


Assuntos
Levanogestrel , Espectrometria de Massas em Tandem , Feminino , Humanos , Cromatografia Líquida , Projetos Piloto , Acetato de Medroxiprogesterona , Anticoncepcionais Orais Combinados
4.
Antivir Ther ; 28(1): 13596535221150746, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36609161

RESUMO

BACKGROUND: Limited data exist for dosing of zanamivir in the setting of CVVH in the intensive care unit (ICU). Our objective is to report the pharmacokinetics and sieving coefficient (Sv) of zanamivir in patients receiving continuous venovenous hemofiltration (CVVH). METHODS: In this prospective observational study, patients of ≥18 years admitted to the ICU with a life-threatening Influenza A or B infection, treated with zanamivir i.v. undergoing CVVH were included. Patients received a zanamivir loading dose of 600 mg i.v., 12 h later followed by maintenance dosages two times daily according to the treating physician. Per patient, nine CFT plasma and nine ultrafiltrate samples were drawn on day 2 of treatment and analysed with a validated HPLC-MS/MS method. RESULTS: Four patients were included in the study. The zanamivir elimination half-life was prolonged with 5.6-9.9 h, compared to patients with normal renal function. A Sv of approximately 1.0 was identified, with unrestricted transport of zanamivir to the ultrafiltrate. CONCLUSIONS: Zanamivir is well cleared by CVVH. In absence of the possibility for therapeutic drug monitoring, the ultrafiltration rate seems as a good surrogate parameter to estimate the CLCVVH and may help guide the dosing of zanamivir.


Assuntos
Terapia de Substituição Renal Contínua , Hemofiltração , Humanos , Zanamivir/uso terapêutico , Hemofiltração/métodos , Estado Terminal/terapia , Espectrometria de Massas em Tandem
5.
Microb Cell Fact ; 22(1): 4, 2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36609265

RESUMO

The metabolic potency of fungi as camptothecin producer elevates their prospective use as an industrial platform for commercial production, however, the loss of camptothecin productivity by fungi with the storage and subculturing are the major obstacle. Thus, screening for endophytic fungal isolates inhabiting ethnopharmacological plants with an obvious metabolic stability and sustainability for camptothecin biosynthesis could be one of the most feasible paradigms. Aspergillus terreus ON908494.1, an endophyte of Cestrum parqui was morphologically and molecularly verified, displaying the most potent camptothecin biosynthetic potency. The chemical identity of A. terreus camptothecin was confirmed from the HPLC, FTIR and LC-MS/MS analyses, gave the same molecular structure and mass fragmentation patterns of authentic one. The purified putative camptothecin displayed a strong anticancer activity towards HepG-2 and MCF-7 with IC50 values 0.96 and 1.4 µM, respectively, with no toxicity to OEC normal cells. As well as, the purified camptothecin displayed a significant antifungal activity towards fungal human pathogen Candida albicans, Aspergillus flavus, and A. parasiticus, ensuring the unique structural activity relationships of A. terreus camptothecin, as a powerful dually active anticancer and antimicrobial agent. The camptothecin productivity of A. terreus was maximized by bioprocessing with Plackett-Burman design, with an overall 1.5 folds increment (170.5 µg/L), comparing to control culture. So, the optimal medium components for maximum yield of camptothecin by A. terreus was acid why (2.0 mL/L), Diaion HP20 (2.0 g/L), Amberlite XAD (2.0 g/L), dextrin (5.0 g/L), glucose (10.0 g/L), salicylic acid (2.0 g/L), serine (4.0 g/L), cysteine (4.0 g/L) and glutamate (10.0 g/L), at pH 6 for 15 days incubation. By the 5th generation of A. terreus, the camptothecin yield was reduced by 60%, comparing to zero culture. Interestingly, the productivity of camptothecin by A. terreus has been completely restored and over increased (210 µg/L), comparing to the 3rd generation A. terreus (90 µg/L) upon addition of methanolic extracts of Citrus limonum peels, revealing the presence of some chemical signals that triggers the camptothecin biosynthetic machinery. The feasibility of complete restoring of camptothecin biosynthetic-machinery of A. terreus for stable and sustainable production of camptothecin, pave the way for using this fungal isolate as new platform for scaling-up the camptothecin production.


Assuntos
Camptotecina , Cestrum , Humanos , Camptotecina/farmacologia , Camptotecina/metabolismo , Endófitos/metabolismo , Cromatografia Líquida , Espectrometria de Massas em Tandem
6.
Environ Monit Assess ; 195(2): 277, 2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36609771

RESUMO

Pesticides are the potent agrochemicals used to successfully manage, repel, or stop pests and weeds in agricultural production. This study analyzed 222 pesticide active substances in 90 samples of seven different vegetables and fruits acquired from producers through liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) and gas chromatography-mass spectrometry/mass spectrometry (GC-MS/MS) technology. The validation parameters of each pesticide's active substances were determined. The LOD, LOQ values, and recovery studies of the 222 active substances were 3.00, 10.00 ng/g, and between 76.07 and 108.08%, respectively. The correlation coefficients and measurement uncertainty were determined to be between 0.990-0.999 and 8.91-31.46%, respectively. There were active substances of chlorpyrifos, acetamiprid, azoxystrobin, difenoconazole, malathion, dieldrin, boscalid, triticonazole, tebuconazole, triadimenol, trifloxystrobin, pirimicarb, and dodine among the vegetable and fruit samples used in the study. There were no active substances in 55 (61%) samples. Among the 35 samples (39%), 31 samples (34%) contained only one active substance, whereas four (5%) contained two active substances. However, the amount of active substances in six (7%) samples was above the maximum residue levels (MRL) limits. Various processes used in the study revealed that peeling was the most effective pesticide residue removal strategy. The washing procedure also proved that it reduced some pesticide residues but failed to eliminate all pesticides. The peeling process successfully reduced a significant amount of the active substances from the products, however, residues remained. Washing the fruits with hot water was also effective in removing residues. As a result, analyses of the peeled sections yielded higher pesticide residue concentrations than those of the entire product.


Assuntos
Resíduos de Praguicidas , Praguicidas , Resíduos de Praguicidas/análise , Espectrometria de Massas em Tandem/métodos , Frutas/química , Verduras/química , Cromatografia Líquida/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Monitoramento Ambiental , Praguicidas/análise , Contaminação de Alimentos/análise
7.
Curr Protoc ; 3(1): e644, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36622836

RESUMO

Accurate identification and quantification of drugs and their metabolites (analytes) in biological matrices is an analytical foundation of clinical and forensic toxicology. For decades, liquid chromatography interfaced by electrospray ionization with tandem mass spectrometry (LC-ESI-MS/MS) has been a widely used technology for analysis in the field of toxicology, as well as in many other fields of bioscience. It is also known that ion response in LC-ESI-MS/MS analysis is influenced by coeluting biological compounds and that preanalytical sample clean-up is often insufficient in removing these interferences. As a result, a normalization technique is commonly used for assessment and compensation of matrix effects encountered in routine analysis. Internal standardization with a stable isotope analog of the analyte is the predominant normalization technique used in LC-ESI-MS/MS analysis. The technique, however, requires commercial availability or costly custom synthesis of an isotopic analog specific for each analyte. Here we describe an alternative technique for matrix normalization for use in high-volume, multianalyte testing without the need for isotope analogs. The technique involves analysis of the original sample (neat analysis) followed by analysis of a second sample aliquot (spike analysis) that has been fortified with a controlled amount of reference analyte. A calibration procedure similar to internal standardization is employed, using an ion response ratio of neat to fortified analyte. As a demonstration of the technique in multianalyte testing, we provide a detailed protocol for simultaneous detection and quantification of 102 drugs and drug metabolites in human urine. We also provide a support protocol for addition of new analytes to the multianalyte panel, allowing convenient collection of the validation data during routine testing. The matrix normalization technique and testing principles may be applicable to a wide range of analytes and biological matrices, not only those encountered in toxicology but also in other fields of bioscience. © 2023 Wiley Periodicals LLC. Basic Protocol: Detection and quantification of 102 toxicology analytes in urine by LC-ESI-MS/MS analysis using the threshold accurate calibration technique Support Protocol: Method for addition and validation of new analytes to expand the Basic Protocol.


Assuntos
Isótopos , Espectrometria de Massas em Tandem , Humanos , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida/métodos , Padrões de Referência , Toxicologia Forense/métodos
8.
Food Res Int ; 163: 112041, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36596086

RESUMO

The purpose was to investigate the contents of heat-induced hazards by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in 44 commercial nuts. Results showed that content ranges of Acrylamide (AA), 5-hydroxymethylfurfural (5-HMF), Nε-carboxymethyl-lysine (CML), Nε-carboxyethyl-lysine (CEL), 3-Deoxyglucosone (3-DG), Glyoxal (GO), and Methylglyoxal (MGO) were ND-123.57 µg/kg, 0.57-213.42 mg/kg, 3.18-18.67 mg/kg, 3.98-57.85 mg/kg, 1.5-133.86 mg/kg, 0.45-1.59 mg/kg and 0.29-13.84 mg/kg, respectively. Sunflower seeds contained more heat-induced hazards followed by pistachios, cashews, almonds, walnuts and hazelnuts. The content of 5-HMF was positively correlated with the content of 3-DG. CML exhibited positive correlation with content of GO while no correlation between CEL and MGO. Higher levels of 3-DG and 5-HMF were observed in nuts produced with sugar and honey. Deep processing had a stronger promoting effect on CML and CEL formation. These data could provide a crucial guide for consumers to select nut products which might reduce heat-induced hazards intake.


Assuntos
Nozes , Espectrometria de Massas em Tandem , Cromatografia Líquida/métodos , Nozes/química , Temperatura Alta , Óxido de Magnésio , Aldeído Pirúvico/análise , Glioxal
9.
Food Res Int ; 163: 112120, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36596088

RESUMO

The freeze-dried (FD) edible roses with high content of bioactive substances and superior flavor have been favored by consumers. Nevertheless, the development of freeze-dried rose industry has been plagued by a long drying time and low efficiency. This study investigated the effects of ultrasonic pretreatment (UP) in multi-frequency modes and electro-infrared pretreatment (EIP) prior to FD on polyphenol accumulation and drying characteristics of roses. The mechanism was explored by the changes in microstructure, equivalent circuit parameters, and phenol identifications of rose. The results showed that the FD time of roses decreased by 26 % after ultrasonic-infrared sequential synergistic pretreatment (UP + EIP) due to the damage of cell membrane permeability from UP. The quality attributes of UP + EIP products including color, phenols, and antioxidant activity (DPPH and ABTS radical scavenging rates) remarkably improved. UP + EIP significantly (p < 0.05) increased the content of polyphenols, namely quercetin-3ß-d-glucoside, phlorizin, procyanidin B2, gallicacid, and rutin in the FD roses quantified by ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-qTOF-MS/MS). Therefore, UP + EIP is an effective pretreatment method for shortening FD time and producing high-quality FD rose products with enhanced polyphenol content.


Assuntos
Polifenóis , Rosa , Espectrometria de Massas em Tandem , Antioxidantes/análise , Dessecação/métodos , Fenóis/análise
10.
Food Res Int ; 163: 112122, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36596090

RESUMO

Durian is a nutritious tropical fruit with potent antioxidant, anti-inflammatory, antibacterial and anti-cancer effects. However, the durian shell was mainly discarded as waste, while there were few studies on the characterization of its phenolic profiles, antioxidant activities, and in vivo metabolites. In the present study, a total of 17 compounds were identified in durian shell extract (DSE) by using an ultra-high-performance liquid chromatography coupled with linear ion trap quadrupole Orbitrap mass spectrometry (UHPLC-LTQ-Orbitrap-MS/MS), while 33 metabolites were found in rats' plasma, urine and organ. Moreover, DSE could effectively reduce H2O2-induced oxidative damage in HepG2 cells, reduce the expression of Reactive Oxygen Species (ROS), Malondialdehyde (MDA) and Lactate Dehydrogenase (LDH) and inhibit apoptosis by regulating the expression of Bcl-2-Associated X (BAX), B-Cell Lymphoma 2 (BCL-2), Caspase-3 and Caspase-9 genes and proteins related to mitochondrial pathway apoptosis. This is the first comprehensive report on Durian shell phenolics, their metabolic profiles and underlying mechanisms of the in vitro antioxidant activities.


Assuntos
Antioxidantes , Bombacaceae , Ratos , Animais , Humanos , Antioxidantes/análise , Bombacaceae/química , Peróxido de Hidrogênio/metabolismo , Espectrometria de Massas em Tandem , Células Hep G2 , Fenóis/química , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo
11.
Food Res Int ; 163: 112170, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36596116

RESUMO

α-dicarbonyl compounds (α-DCs) serve as potential biomarkers for oxidative stress-related diseases but are difficult to detect.To study the metabolism of carbonyl compounds, we developed a new mass spectrometry probe, 3-benzyl-2-oxo-4λ3-thiazolidine-4-carbohydrazide (BOTC), containing hydrazyl groups for the targeted detection of carbonyl functional groups.In a novel approach, we used BOTC pre-column derivatization with ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to simultaneously detect four kinds of α-DCs in red wine as well as in urine after drinking. The α-DCs were completely separated (R2 ≥ 0.9995), detection was sensitive (detection limit was 12.5-50 fmol), consistent (intraday and interday precision was 0.1-5.7 %), and efficient (average recoveries were 103.3-110.2 %). The method was applied to the analysis of α-DCs in different wines and the dynamic monitoring of transit and excretion in vivo after drinking. Our novel method provides a new strategy for the detection of α-dicarbonyl compounds in red wine and dicarbonyl compounds produced in oxidative stress-related diseases.


Assuntos
Espectrometria de Massas em Tandem , Vinho , Humanos , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas em Tandem/métodos , Vinho/análise
12.
Food Res Int ; 163: 112187, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36596128

RESUMO

Quinones are electrophilic compounds that can undergo Michael addition or Schiff base reaction with nucleophilic amines, but the effect of temperature has not been systematically studied. The aim of this study was to characterize how temperature affects the reaction mechanism and kinetics of 4-methylbenzoquinone (4MBQ) with lysine (Lys), Nα-acetyl Lys or Nε-acetyl Lys. The products were identified and characterized by LC-MS/MS, which revealed formation of Michael addition products, Schiff base, and a di-adduct in Lys and Nα-acetyl Lys-containing reaction mixtures. The product profiles were not affected by temperature in the range of 15-100 °C. NMR analysis proved that Michael addition of Nα-acetyl Lys occurred on the C5 position of 4MBQ. Rate constants for the reactions studied by stopped-flow UV-vis spectrophotometry under pseudo-first-order conditions where the amines were present in excess in the range 15 °C to 45 °C showed the α-amino groups of Lys are more reactive than the ε-groups. The kinetics results revealed that the temperature dependence of reaction rates followed the Arrhenius law, with activation energies in the order: Lys < Nε-acetyl Lys < Nα-acetyl Lys. Our results provide detailed knowledge about the temperature dependence of the reaction between Lys residues and quinones under conditions relevant for storage of foods.


Assuntos
Lisina , Bases de Schiff , Lisina/química , Cromatografia Líquida , Temperatura , Espectrometria de Massas em Tandem , Aminas , Quinonas
13.
Food Res Int ; 163: 112262, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36596173

RESUMO

Sea cucumber tentacles (aquapharyngeal bulb) are a rich source of biologically active compounds, including phenolics, however they are mainly discarded as processing waste. This study evaluated free, esterified, and insoluble-bound phenolics of Atlantic sea cucumber (Cucumaria frondosa) tentacles and their antioxidant activity for the first time. Biological properties such as inhibitory activities against α-glucosidase, tyrosinase, and the formation of AGEs as well as LDL-cholesterol and DNA oxidation were investigated. The antioxidant activity of the phenolic extracts was also evaluated in a fish model system. In the UHPLC-QTOF-MS/MS analysis, 31 phenolic compounds, mainly phenolic acids and flavonoids, were identified and quantified. Among them, eight compounds were detected for the first time in any species of sea cucumber. The free phenolic fraction was the major form of phenolics, mainly protocatechuic acid, p-coumaric acid, gallic acid, ellagic acid, catechin, and quercetin, exhibiting strong antioxidant and biological activities. Fresh Atlantic salmon treated with sea cucumber phenolics delayed lipid oxidation as measured by the thiobarbituric acid reactive substances (TBARS) assay. Therefore, Atlantic sea cucumber tentacles may serve as a viable source of functional food ingredients with protective antioxidant properties.


Assuntos
Cucumaria , Pepinos-do-Mar , Animais , Antioxidantes/farmacologia , Antioxidantes/análise , Espectrometria de Massas em Tandem , Fenóis/análise
14.
Food Res Int ; 163: 112290, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36596195

RESUMO

Ambelania duckei Markgr is a species of the Apocynaceae family, native to the Amazon region that is unexplored from a nutritional point of view and studied in relation to its chemical constituents. This work presents an unprecedented study of the proximate composition, lipid profile, a chromatographic analysis, and the antioxidant activity of extracts obtained from the pulp, peel and seeds of the fruit. The results showed that potassium, calcium, and magnesium stood out as the most abundant key minerals in the fruit peel and pulp, with an emphasis on the potassium present in the fruit pulp at 1750.0 mg/100 g. The peel had the highest content of total phenolics (374.86 mg/g), flavonoids (15.54 mg/g), tannins (27.45 mg/g) and O-diphenols (379.36 mg/g; 645.71 mg/g). The antioxidant activity (AA) was highest in the peel compared to the pulp in the DPPH, ABTS, and ORAC tests showing: IC50 of 29.82; 43.67; and 407.13 µg/mL, respectively but a lower activity for the Fe2+ chelator. The analysis of the lipid fractions from the peel, pulp, and seeds of the A. duckei fruit resulted in 14 types of fatty acids. The major fatty acids found in the three parts of the fruit were oleic acid (peel, 22.52 %), palmitic acid (pulp, 17.34 %), and linoleic acid (seeds, 47.99 %). The lipid profile and nutritional aspects had a PUFA/SFA ratio (0.4-1.8) in the different parts of the A. duckei fruit; the atherogenic and thrombogenic indexes were higher in the peel (1.23) and pulp (0.62), respectively. The ratio between the hypocholesterolemic and hypercholesterolemic fatty acids (0.5 - 3.8) calculated for the fruit are within the desirable range for a nutritious food. The chromatographic analysis of the volatile organic compounds (VOCs) from the peel and pulp of the fruit, identified 74 VOCs, of which 60.9 % are related to terpenes, and emit notes such as cucumber, green, fatty, floral, and mint, due to the presence of substances with OAVs > 10, especially α-ionone, 1,8-cineole, 2,4-decadienal, and dodecanal. The analysis of the MS and MS/MS spectra of the chromatograms obtained by LC- QTOF-HRMS led to the identification of 26 compounds in the peel, seeds and pulp of A. duckei, such as fatty acids, phenolic acid, flavonoids, proanthocyanidins, alkaloids, and terpenoids. The results show that the pulp of A. duckei has potential as nourishing food and the nutritional and chemical aspects of the peel can be applied to commercial applications.


Assuntos
Apocynaceae , Frutas , Frutas/química , Antioxidantes/análise , Espectrometria de Massas em Tandem , Flavonoides/análise , Ácidos Graxos/análise , Terpenos/análise , Potássio/análise
15.
Food Res Int ; 163: 112315, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36596206

RESUMO

LC-HR-MS/MS is the predominant analytical technique in phenolic compound (PC) research. However, the manual interpretation of mass spectra is a heavy nontrivial time-consuming task and depends on mass spectrometry and phenolic compounds fragmentation deep knowledge. We think this manual approach should be partially translated into a practical software that allows users to perform such complicated analyses. In silico fragmentation software have been tested for small molecule identification, MS-FINDER and SIRIUS stood out at identification contests and challenges. We evaluated both software to identify PC from two data categories: 1st MS/MS spectra from 18 phenolic compound standards (PCS) and 2nd phenolic compounds from 8 food samples (FPC) (coffee, green tea, cranberry juice, grape juice, orange juice, apple juice, soy extract and parsley extract). MS-FINDER and SIRIUS were able to correctly identifymore than 90% of the PCS by LC-HR-MS/MS. The main FPC were also correctly identified by MS-FINDER (70%) and SIRIUS (38%). We highlight that these software were unable to differentiate PC isomers. This task is only possible by using additional information, such as chromatographic behavior and manual analysis of the relative intensity of fragments in the MS/MS spectra. Therefore, the combination of initial screening by using MS-FINDER and SIRIUS with manual analyses of additional information is a powerful and efficient approach for identifying phenolic compounds.


Assuntos
Fenóis , Espectrometria de Massas em Tandem , Espectrometria de Massas em Tandem/métodos , Fenóis/análise , Cromatografia Líquida/métodos , Café , Extratos Vegetais
16.
Chem Pharm Bull (Tokyo) ; 71(1): 31-40, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36596510

RESUMO

Intercellular lipids fill the interstices of corneocytes and serve a barrier function. The amount of transdermal water evaporation varies depending on the packing structure of intercellular lipids, as this structure is important for maintaining barrier efficacy. This packing structure consists of a mixture of crystals (orthorhombic and hexagonal) and liquid crystals (fluid phase), and the proportion of these phases is thought to affect barrier function. However, there have been no methods to visualize the actual distribution of the domains formed by packing structure in intercellular lipids. In this study, the planar distribution of intercellular lipid structures was determined using focal plane array (FPA)-based Fourier transform (FT) IR imaging analysis of stratum corneum cell units obtained by grid stripping. The lipid composition of ceramides was revealed by electrospray ionization tandem mass spectrometry (ESI-MS/MS)-based shotgun lipidomics. The distribution of domains formed by packing structures and the lipid composition of ceramides was compared in skin with high- or low-transepidermal water loss (TEWL). The orthorhombic proportion was lower in high-TEWL skin than in low-TEWL skin. ESI-MS/MS-based shotgun lipidomics analysis showed that the alpha-hydroxyceramide content in the low- and high-TEWL groups differed regarding the distribution of fatty acid chain lengths. The evaluation of stratum corneum cell units using FPA-based FTIR imaging is an innovative technology that can visualize the distribution of domains formed by intercellular lipid-packing structures. Increased proportions of alpha-hydroxyceramide subclasses such as alpha-hydroxy-sphingosine ceramide and alpha-hydroxy-phytosphingosine ceramide were associated with a reduced proportion of the orthorhombic packing structure domain.


Assuntos
Epiderme , Espectrometria de Massas em Tandem , Epiderme/química , Pele/química , Ácidos Graxos , Ceramidas/química , Água/química
17.
Phytomedicine ; 109: 154578, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36610146

RESUMO

BACKGROUND AND PURPOSE: As a complex and challenging complication for the patients with diabetes mellitus, diabetic ulcers are difficult to heal and current strategies cannot fulfill the patients' requirements. Pien Tze Huang (PZH) is a standardized medicine approved for various wounds treatments, and this study systematically investigated the effect and mechanism of intragastric administration of PZH (I-PZH) on diabetic wound healing. METHODS AND RESULTS: The effect of I-PZH on the healing of full-thickness wounds in rats with diabetes mellitus which was induced by high fat diet followed by streptozotocin injection was evaluated, and RNA sequencing (RNA-seq) and targeted central carbon metabolism metabolomics were combined to explore the underlying mechanism. I-PZH promoted wound healing, facilitated extracellular matrix synthesis, and maintained body weight of rats, but did not affect fasting blood glucose levels. Additionally, I-PZH significantly decreased 8-OHdG, cleaved caspase 3 and MMP9 levels, and increased TGF-ß1 expression. RNA-seq analysis showed that I-PZH inhibited inflammation and that the vital common targets were TLR2, IL-17A and IL-1ß; specifically affected "energy derivation by oxidation of organic compounds" with UQCRC1, NDUFS3 and SDHA as vital specific targets. Further experiments confirmed that I-PZH reduced TLR2, IL-17A and IL-1ß, increased UQCRC1, SDHA, NDUFS3, promoted ATP synthesis and restored activity of mitochondrial respiratory chain complexes I and III in diabetic wounds. Metabolomics by HPLC-MS/MS analysis showed that I-PZH reversed multiple energy metabolism-related metabolites such as glucuronic acid, GMP, d-gluconic acid, cis-aconitic acid, ribose 5-phosphate and pantothenate. CONCLUSION: This study highlights the important role of inflammation and energy generation in diabetic wound healing, reveals wound repair mechanism of PZH and promotes its clinical application in diabetic wound treatment.


Assuntos
Diabetes Mellitus , Interleucina-17 , Ratos , Animais , Espectrometria de Massas em Tandem , Receptor 2 Toll-Like , Inflamação , Cicatrização
18.
Phytomedicine ; 109: 154544, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36610155

RESUMO

BACKGROUND: Pinnatifolone A is a typical sesquiterpenoid and the primary active ingredient of Syringa oblata Lindl., has potent anti-inflammatory activity. However, Pinnatifolone A pharmacokinetic and metabolites analysis investigations in male and female rats, as well as its in vitro stability in male and female rat liver microsomes, have not been evaluated and compared. PURPOSE: To investigate preclinical pharmacokinetic and metabolite in both genders, confirm gender differences, and provide usable information for the development of clinical applications. METHODS: A quick, precise, and sensitive LC-MS/MS method was created and effectively used to determine the pharmacokinetics of oral (140 mg/kg) and intravenous (6.3 mg/kg) Pinnatifolone A in male and female rats, in vitro Pinnatifolone A elimination studies in male and female rat liver microsomes. Following that, a UHPLC-Q-TOF-MS/MS technique was established to identify the metabolic profiles of Pinnatifolone A obtained from rat plasma and excreta. RESULTS: In the current study, we established for the first time an LC-MS/MS method for the quantitation of Pinnatifolone A with acceptable linearity and selectivity, recovery and matrix effect, accuracy and precision. The absolute oral bioavailability of Pinnatifolone A was approximately 30.36% in female rats, the clearance (CL) was 20.99±3.33 l/h/kg in female rats and 472.37±437.31 l/h/kg in male rats. This difference in rat genders may pertain to the sex-specific expression of hepatic enzymes as demonstrated in the metabolic stability evaluation in the present research; the male rats exhibited higher CLint(mic) (158.83±9.57 µl/min/mg protein) than female rats (76.47±7.90 µl/min/mg protein) liver microsomes, indicating higher Pinnatifolone A clearance in male rats. Twenty-four metabolites were detected and identified in female and male rats; N-acetylcysteine conjugation metabolite was the most abundant metabolites in both rat feces and urine. Furthermore, male and female rats had significantly different levels of the N-acetylcysteine conjugation metabolite. Hydrogenation metabolite was particular to female rats both in rat fecal and urine. Glucuronide conjugation metabolite was the predominant metabolite in rat plasma, and its amount in female rats was double that of male rats. CONCLUSIONS: The present research is the first to report the preclinical pharmacokinetics and metabolites of Pinnatifolone A in male and female rats, confirming the gender-based differences. The findings provide a comprehensive overview for further understanding of the pharmacokinetic and metabolic characteristics of Pinnatifolone A and serve as a guide for its future development and utilization.


Assuntos
Acetilcisteína , Espectrometria de Massas em Tandem , Ratos , Feminino , Masculino , Animais , Espectrometria de Massas em Tandem/métodos , Disponibilidade Biológica , Cromatografia Líquida , Cromatografia Líquida de Alta Pressão/métodos , Fatores Sexuais , Administração Oral
19.
BMC Genomics ; 24(1): 8, 2023 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-36624393

RESUMO

BACKGROUND: Exosomes are nanosized membranous vesicles secreted by various types of cells, which facilitate intercellular communication by transporting bioactive compounds. Exosomes are abundant in biological fluids including semen, and their protein composition and the potential of seminal plasma exosomes (SPEs) as fertility biomarkers were elucidated in humans, however, little information is available regarding buffalo (Bubalus bubalis). Here, we examined protein correlation between spermatozoa, seminal plasma (SP), and SPEs, and we compared and analyzed protein differences between high-motility (H-motility) and low-motility (L-motility) SPEs in buffalo. RESULTS: SPEs were concentrated and purified by ultracentrifugation combined with sucrose density gradient centrifugation, followed by verification using western blotting, nanoparticle tracking analysis, and transmission electron microscopy. Protein composition in spermatozoa, SP and SPEs, and protein difference in H- and L-motility SPEs were identified by LC-MS/MS proteomic analysis and were functionally analyzed through comprehensive bioinformatics. Many SPEs proteins originated from spermatozoa and SP, and nearly one third were also present in spermatozoa and SP. A series of proteins associated with reproductive processes including sperm capacitation, spermatid differentiation, fertilization, sperm-egg recognition, membrane fusion, and acrosome reaction were integrated in a functional network. Comparative proteomic analyses showed 119 down-regulated and 41 up-regulated proteins in L-motility SPEs, compared with H-motility SPEs. Gene Ontology (GO) enrichment of differentially expressed proteins (DEPs) showed that most differential proteins were located in sperm and vesicles, with activities of hydrolase and metalloproteinase, and were involved in sperm-egg recognition, fertilization, single fertilization, and sperm-zona pellucida binding processes, etc. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that differential proteins were mainly involved in the PPRP signaling pathway, calcium signaling pathway, and cAMP signaling pathway, among others. Furthermore, 6 proteins associated with reproduction were validated by parallel reaction monitoring analysis. CONCLUSION: This study provides a comprehensive description of the seminal plasma exosome proteome and may be of use for further screening of biomarkers associated with male infertility.


Assuntos
Exossomos , Sêmen , Animais , Masculino , Humanos , Sêmen/metabolismo , Búfalos , Motilidade Espermática , Cromatografia Líquida , Exossomos/metabolismo , Proteômica , Espectrometria de Massas em Tandem , Espermatozoides/metabolismo , Proteoma/metabolismo
20.
AAPS PharmSciTech ; 24(1): 32, 2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36627414

RESUMO

Migraine headaches are usually intolerable, and a quick-relief treatment remains an unmet medical need. Almotriptan malate is a serotonin (5-HT1B/1D) receptor agonist approved for the treatment of acute migraine in adults. It is currently available in an oral tablet dosage form and has a Tmax of 1-3 h, and therefore, there is a medical need to develop a non-invasive rapidly acting formulation. We have developed an intranasal formulation of almotriptan malate using the quality-by-design (QbD) approach. A 2-factor 3-level full factorial design was selected to build up the experimental setting. The developed formulation was characterized for pH, viscosity, in vitro permeation, ex vivo permeation, and histopathological tolerance. To assess the potential of the developed formulation to produce a rapid onset of action following intranasal delivery, a pharmacokinetic study was performed in the Sprague-Dawley rat model and compared to the currently available marketed oral tablet formulation. For this, the LC-MS/MS bioanalytical method was developed and used for the determination of plasma almotriptan malate concentrations. Results of a pharmacokinetic study revealed that intranasal administration of optimized almotriptan malate formulation enabled an almost five-fold reduction in Tmax and about seven-fold increase in bioavailability in comparison to the currently available oral tablet formulation, suggesting the potential of developed almotriptan malate intranasal formulation in producing a rapid onset of action as well as enhanced bioavailability.


Assuntos
Transtornos de Enxaqueca , Agonistas do Receptor de Serotonina , Animais , Ratos , Administração Intranasal , Cromatografia Líquida , Agonistas do Receptor de Serotonina/farmacocinética , Ratos Sprague-Dawley , Espectrometria de Massas em Tandem , Triptaminas/farmacocinética , Transtornos de Enxaqueca/tratamento farmacológico , Serotonina/uso terapêutico , Comprimidos
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