Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 60.400
Filtrar
1.
Braz. j. biol ; 83: e246312, 2023. tab, graf
Artigo em Inglês | MEDLINE, LILACS, VETINDEX | ID: biblio-1339357

RESUMO

Abstract The antioxidant, photoprotective and antinociceptive Marcetia macrophylla active extract was investigated as an active ingredient in a sunscreen cream formulation. Thus, the M. macrophylla extract showed IC50 of 3.43 mg/ml of the antioxidant (DPPH∙ scavenging test) and Sun Protection Factor of 20.25 (SPF/UV-B, at 250 µg/ml) and UV-A of 78.09% (photobleaching trans-resveratrol test). The antinociceptive activity was superior to all standards tested using the in vivo acetic acid-induced writhing test (99.14% at the dose of 200 mg/kg) and the high-performance liquid chromatography coupled with diode array detector and mass spectroscopy multi-stage (HPLC-DAD-MS/MS) enabled the structural characterization of the quercetin-3-O-hexoside, quercetin-3-O-pentoside and quercetin-3-O-desoxihexoside. The pharmaceutical formulation containing the Marcetia macrophylla crude active extract was prepared and the physicochemical tests (organoleptic characteristics, pH analysis and centrifugation), the in vitro UVB (sun protection factor, SPF) and UVA (β-carotene) using the spectroscopic method were investigated. The formulation showed satisfactory results concerning the physicochemical parameters evaluated and active against the UV test. Thus, M. macrophylla showed biological activities with potential use in pharmaceutical preparations.


Resumo O extrato bruto de Marcetia macrophylla mostrou atividade antioxidante, fotoprotetora e antinociceptiva, sendo em seguida investigado como ingrediente ativo em uma formulação fotoprotetora. Assim, o extrato de M. macrophylla apresentou atividade antioxidante com IC50 de 3,43 mg/mL (teste de sequestro do DPPH∙) e Fator de Proteção Solar de 20,25 (FPS/UV-B, 250 µg/mL) e UV-A de 78,09% (teste de fotobranqueamento do trans-resveratrol). A atividade antinociceptiva usando o teste in vivo de contorções abdominais induzidas por ácido acético foi superior a todos os padrões testados (99,14% na dose de 200 mg/Kg). A análise por cromatografia líquida de alta eficiência acoplada a detector de fotodiodos e espectroscopia de massas multi-estágio (CLAE-DAD-EM/EM) possibilitou a caracterização dos flavonoides quercetina-3-O-hexosídeo, quercetina-3-O-pentosídeo e quercetina-3-O-desoxihexosídeo. A formulação farmacêutica contendo o extrato ativo bruto de Marcetia macrophylla foi preparada e os testes físico-químicos (características organolépticas, análise de pH e centrifugação), o UVB in vitro (fator de proteção solar, FPS) e UVA (β-caroteno) foram investigados. A formulação apresentou resultados satisfatórios frente aos parâmetros físico-químicos avaliados e ativos contra UV. Assim, M. macrophylla apresentou atividades biológicas com potencial uso em preparações fitofarmacêuticas.


Assuntos
Protetores Solares/farmacologia , Antioxidantes/farmacologia , Extratos Vegetais/farmacologia , Espectrometria de Massas em Tandem , Analgésicos/farmacologia
2.
Braz. j. biol ; 83: e248083, 2023. tab, graf
Artigo em Inglês | MEDLINE, LILACS, VETINDEX | ID: biblio-1278546

RESUMO

Abstract Species of the genus Cordia have shown biological activities, such as anti-inflammatory, analgesic, antioxidant, antiviral, and antifungal activities. The species Cordia glabrata (MART) A.DC. Has no information concerning its phytochemical profile and possible biological activities. Thus, this study aimed to evaluate this profile in ethanolic extracts of young, adult and senescent leaves, as well as their antioxidant, photoprotective, antimicrobial, and virucidal potentials. Phytochemical analysis was performed by TLC (thin-layer chromatography) and showed the presence of flavonoids, tannins, and terpenes. The evaluation by UPLC-MS/MS (Ultra performance liquid chromatography - tandem mass spectrometer) evidenced the presence of caffeic (3.89 mgL-1), p-cumaric (6.13 mgL-1), and ferulic (0.58 mgL-1) acids, whilst, in GC/MS (Gas chromatography-mass spectrometry) analysis there was a greater amount of palmitic (51.17%), stearic (20.34%), linoleic (9.62%), and miristic (8.16%) fatty acids. The DPPH (2,2-Diphenyl-1-picrylhydrazyl) and ABTS+ (2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radicals were used to verify the potential antioxidant activity, observing a better activity for the leaf extract in the adult phenological stage: 54.63 ± 1.06 µgmL-1 (DPPH) and 44.21 ± 1.69 mM (ABTS). The potential photoprotective activity of the extracts was determined by spectrophotometry and the in vitro values of SPF (Sun Protection Factor) in young and adult leaves (5.47 and 5.41, respectively) showed values close to the minimum SPF of 6.0 required by ANVISA (Brazilian Health Regulatory Agency). It was not observed an antimicrobial activity for Staphylococcus aureus with a minimum inhibitory concentration of 2000 μgmL-1, however the anti-herpetic assay against the Herpes simplex virus type 2 (HSV-2) showed a potent virucidal activity at the tested concentrations with CV50 value <0.195 μgmL-1 and a Selectivity Index (SI = CC50 / CV50) greater than 448. The results obtained in this study suggest that extracts of leaves of C. glabrata in their adult phenological stage have potential antioxidant, photoprotective and virucidal activity, considering in vitro test results.


Resumo Espécies do gênero Cordia apresentam atividades biológicas, como anti-inflamatória, analgésica, antioxidante, antiviral e antifúngica. Para a espécie Cordia glabrata (MART) A.DC., ainda não existem informações sobre seu perfil fitoquímico e possíveis atividades biológicas, deste modo, o presente estudo teve como objetivo avaliar este perfil em extratos etanólicos de folhas jovens, adultas e senescentes, bem como o potencial antioxidante, fotoprotetor, antimicrobiano e virucida. A análise fitoquímica foi realizada por CCD (Cromatografia em Camada Delgada), mostrando a presença de flavonóides, taninos e terpenos. Na avaliação por CLAE EM/EM (Cromatografia Líquida de Ultra Eficiência acoplada a Espectrometria de Massas) foi evidenciado a presença dos ácidos caféico (3,89 mgL-1), p-cumárico (6,13 mgL-1) e ferúlico (0,58 mgL-1), paralelamente, na CG/EM (Cromatografia Gasosa acoplada a Espectrometria de Massas) verificou-se maior quantidade dos ácidos graxos palmítico (51,17%), esteárico (20,34%), linoléico (9,62%) e mirístico (8,16%). Os radicais DPPH (2,2-Difenil-1-picrilhidrazil) e ABTS+ (2′-Azino-bis (ácido 3-etilbenzotiazolina-6-sulfônico)) foram utilizados para verificar o potencial antioxidante, observando-se uma atividade superior para o extrato da folha em sua fase fenológica adulta: 54,63 ± 1,06 µgmL-1 (DPPH) e 44,21 ± 1,69 mM (ABTS+). A potencial atividade fotoprotetora dos extratos foi determinada espectrofotometricamente e os valores in vitro de FPS (Fator de Proteção Solar) em folhas jovens e adultas (5,47 e 5,41 respectivamente) apresentaram valores próximos ao FPS mínimo de 6,0 exigido pela ANVISA (Agência Nacional de Vigilância Sanitária). Não foi observada atividade antimicrobiana para Staphylococcus aureus sendo a concentração inibitória mínima de 2000 μgmL-1, no entanto o ensaio anti-herpético contra o vírus Herpes simplex tipo 2 (HSV-2) mostrou uma potente atividade virucida nas concentrações testadas com um valor de CV50 <0,195 μgmL-1 e um Índice de Seletividade (IS = CC50 / CV50) maior que 448. Os resultados obtidos neste estudo sugerem que extratos de folhas de C. glabrata em seu estágio fenológico adulto apresentam potencial antioxidante, fotoprotetora e virucida, considerando os resultados de testes in vitro.


Assuntos
Cordia , Anti-Infecciosos , Antivirais/farmacologia , Brasil , Extratos Vegetais/farmacologia , Cromatografia Líquida , Folhas de Planta , Espectrometria de Massas em Tandem , Antioxidantes/farmacologia
3.
J Ethnopharmacol ; 291: 115145, 2022 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-35219821

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Shenling Baizhu San (SBS) is commonly employed to improve gastrointestinal dysfunction in patients with ulcerative colitis (UC) in China. SBS combined with mesalamine has been demonstrated to result in improve its curative effects without increasing any adverse reactions, but the underlying mechanism remains unclarified. AIM OF THE STUDY: Our study aimed to illuminate the potential therapeutic effects and mechanisms of SBS, which is a medicine complementary to mesalamine, in the treatment of UC. MATERIALS AND METHODS: A prospective cohort study was conducted to evaluate the efficacy of SBS as a complementary medicine to mesalamine for patients with UC (n = 48). The patients in the control group (n = 24) were given mesalamine alone, whereas those in the experimental group were administered mesalamine combined with SBS. The therapeutic outcome was assessed at 8 weeks. The structures of the gut microbiota (GMB) were characterized by 16S rRNA sequencing, and the microbial tryptophan metabolites were analyzed by UPLC-MS/MS to investigate the mechanism through which SBS achieves its effects. RESULTS: Our results showed that the combination of SBS and mesalamine could significantly improve the clinical signs of UC by achieving mucosal healing and relieving colon damage. Interestingly, the combination of SBS and mesalamine could alter the GMB structures and increase the microbial levels of tryptophan metabolites, including indole-3-propionic acid and indole-3-acetic acid. CONCLUSION: SBS combined with mesalamine is effective in improving the clinical and endoscopic outcomes of patients with UC. SBS, as a complementary therapy to conventional treatment, alleviates UC via the GMB-tryptophan metabolite axis.


Assuntos
Colite Ulcerativa , Terapias Complementares , Microbioma Gastrointestinal , Cromatografia Líquida , Colite Ulcerativa/tratamento farmacológico , Colite Ulcerativa/metabolismo , Medicamentos de Ervas Chinesas , Humanos , Mesalamina/farmacologia , Mesalamina/uso terapêutico , Estudos Prospectivos , RNA Ribossômico 16S , Espectrometria de Massas em Tandem , Triptofano
4.
Bioanalysis ; 14(5): 267-278, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35195037

RESUMO

Background: The degree of human hepatocyte replacement in chimeric mice with humanized liver has previously been shown to correlate with human plasma albumin measurements. However, there are no reports that directly compare the remaining endogenous mouse albumin with the newly expressed human albumin following engraftment. To better understand the disposition of serum albumin in PXB-mice, we developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to simultaneously quantitate both human and mouse albumin from plasma. Results: A robust correlation was observed between the serum human albumin levels measured by LC-MS/MS and the estimated replacement index of PXB-mice. Conclusion: All data were shown to be specific and suitable to accurately quantify both human and mouse albumin from plasma of chimeric mice with humanized livers.


Assuntos
Albumina Sérica Humana , Espectrometria de Massas em Tandem , Quimera , Cromatografia Líquida , Hepatócitos , Humanos , Fígado , Espectrometria de Massas em Tandem/métodos
5.
Bioanalysis ; 14(5): 307-316, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35195039

RESUMO

Aim: HCP002, a phosphate-modified derivative of voriconazole, can improve solubility without using the nephrotoxic solubilizer, sulfobutylether-ß-cyclodextrin. To study pharmacokinetics in humans, LC-MS/MS methods to quantify HCP002 in human plasma and urine were developed and validated. Method: After protein precipitation by acetonitrile containing voriconazole-d3, HCP002 was separated on a ZORBAX SB-Aq column, and LCMS/MS analysis was performed in multi-response monitoring mode. Results: The analytical run time was 3 min. Linearity was observed over the ranges of 0.100-40.0 and 0.400-200 µg/ml in plasma and urine, respectively. Precision and accuracy were within acceptable limits. Sample stability was confirmed. Conclusion: Rapid and reproducible methods quantified HCP002 in urine, and plasma samples were established.


Assuntos
Plasma , Espectrometria de Massas em Tandem , Cromatografia Líquida/métodos , Humanos , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem/métodos
6.
Bioanalysis ; 14(5): 279-287, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35188423

RESUMO

Background: Currently, no regulatory guidelines are available for parallelism assessment for LC-MS biomarker quantification. Spike recovery, standard addition and dilutional linearity are recommended with no mention of the implications of applying these approaches. Results: Here, using human urine creatinine, the authors compared spike recovery and standard addition in LC-MS biomarker quantification, and evaluated a new hybrid approach: parallelism QCs. The authors drew different conclusions based on which approach was used (<15% cutoff). Conclusion: Current recommended approaches may lead to different conclusions and are not equivalent and interchangeable. The authors recommend that standard addition should be the universal 'go-to' method for LC-MS biomarker parallelism assessment; parallelism QCs, which consider the total concentration as the theoretical value, can be used if the authentic matrix is limited.


Assuntos
Espectrometria de Massas em Tandem , Biomarcadores , Cromatografia Líquida/métodos , Creatinina , Humanos , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem/métodos
7.
Cell Mol Life Sci ; 79(6): 336, 2022 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-35657417

RESUMO

BACKGROUND: Multiple system atrophy (MSA) is a rare, progressive, neurodegenerative disorder presenting glia pathology. Still, disease etiology and pathophysiology are unknown, but neuro-inflammation and vascular disruption may be contributing factors to the disease progression. Here, we performed an ex vivo deep proteome profiling of the prefrontal cortex of MSA patients to reveal disease-relevant molecular neuropathological processes. Observations were validated in plasma and cerebrospinal fluid (CSF) of novel cross-sectional patient cohorts. METHODS: Brains from 45 MSA patients and 30 normal controls (CTRLs) were included. Brain samples were homogenized and trypsinized for peptide formation and analyzed by high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS). Results were supplemented by western blotting, immuno-capture, tissue clearing and 3D imaging, immunohistochemistry and immunofluorescence. Subsequent measurements of glial fibrillary acid protein (GFAP) and neuro-filament light chain (NFL) levels were performed by immunoblotting in plasma of 20 MSA patients and 20 CTRLs. Finally, we performed a proteome profiling of 144 CSF samples from MSA and CTRLs, as well as other parkinsonian disorders. Data were analyzed using relevant parametric and non-parametric two-sample tests or linear regression tests followed by post hoc tests corrected for multiple testing. Additionally, high-throughput bioinformatic analyses were applied. RESULTS: We quantified more than 4,000 proteins across samples and identified 49 differentially expressed proteins with significantly different abundances in MSA patients compared with CTRLs. Pathway analyses showed enrichment of processes related to fibrinolysis and complement cascade activation. Increased fibrinogen subunit ß (FGB) protein levels were further verified, and we identified an enriched recognition of FGB by IgGs as well as intra-parenchymal accumulation around blood vessels. We corroborated blood-brain barrier leakage by a significant increase in GFAP and NFL plasma levels in MSA patients that correlated to disease severity and/or duration. Proteome profiling of CSF samples acquired during the disease course, confirmed increased total fibrinogen levels and immune-related components in the soluble fraction of MSA patients. This was also true for the other atypical parkinsonian disorders, dementia with Lewy bodies and progressive supra-nuclear palsy, but not for Parkinson's disease patients. CONCLUSION: Our results implicate activation of the fibrinolytic cascade and immune system in the brain as contributing factors in MSA associated with a more severe disease course.


Assuntos
Atrofia de Múltiplos Sistemas , Doença de Parkinson , Transtornos Parkinsonianos , Encéfalo/metabolismo , Cromatografia Líquida , Estudos Transversais , Progressão da Doença , Fibrinogênio/metabolismo , Proteína Glial Fibrilar Ácida/metabolismo , Humanos , Atrofia de Múltiplos Sistemas/metabolismo , Doença de Parkinson/metabolismo , Transtornos Parkinsonianos/metabolismo , Transtornos Parkinsonianos/patologia , Proteoma/metabolismo , Espectrometria de Massas em Tandem
8.
Sci Total Environ ; 838(Pt 3): 156391, 2022 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-35654199

RESUMO

With the ever-increasing plastic pollution, the nano-sized plastic particles that are constantly released from the main materials have a greater potential threat. Studies continue on how to eliminate plastic waste, which has become a global problem, from nature. We are aware that complete elimination is not easy at all, but it is not known clearly that even if it is successful, its effects on organisms will also disappear completely. In this study, zebrafish injected with 20 nm-sized polystyrene particles (PS) only during the embryonic period were grown in an environment without plastic exposure. The effects of PS on their offspring embryo/larvae were examined at morphological, molecular and metabolomic levels. Results showed that parental PNP exposure caused significant malformations, decreased survival rate, increased heart rate and blood flow rate, as well as decreased eye size, height and locomotor activity, which were attributed to growth retardation in the offspring. According to the results of whole-mount immunofluorescence larval staining, cell death and reactive oxygen species were significantly increased, while lipid accumulation was decreased in new generation larvae from zebrafish injected with PNP. In order to elucidate the mechanisms underlying these morphological, physiological and molecular damages, the metabolome analyses were performed by evaluating the Q-TOF MS/MS spectra with chemometric analyses in the offspring larvae. According to the metabolomics results, 28 annotated metabolomes suggested by the OPLS-DA analysis that may vary significantly through a variable in projection scores were detected. In addition, it was detected that the significantly increased histopathological findings and immunopositivity of JNK, H2A.X, PI3 and NOP10 in new generation larvae. In conclusion, it has been shown that exposure to PS, even only during the embryonic period, may affect many cancer-related biological processes in the next generation.


Assuntos
Fenômenos Biológicos , Nanopartículas , Neoplasias , Poluentes Químicos da Água , Animais , Larva , Nanopartículas/metabolismo , Plásticos/metabolismo , Poliestirenos/metabolismo , Poliestirenos/toxicidade , Espectrometria de Massas em Tandem , Poluentes Químicos da Água/metabolismo , Peixe-Zebra/metabolismo
9.
J Ethnopharmacol ; 295: 115427, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-35654350

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Danhe granule (DHG) is used by Chinese doctors to treat blood stasis, phlegm and dampness. Its lipid-lowering ability has been investigated in our previous research. However, the anti-liver inflammatory and fibrotic effects and mechanism of action of DHG in non-alcoholic steatohepatitis (NASH) have not been explored. AIM OF THE STUDY: To evaluate the ameliorative effects of DHG on liver inflammation and fibrosis in a methionine/choline-deficient (MCD) diet-induced NASH rat model, and its underlying mechanism. MATERIALS AND METHODS: Sprague-Dawley rats were fed an MCD diet for two weeks and then treated with or without DHG by oral gavage for eight weeks. Their body weight and liver index were measured. The serum alanine aminotransferase (ALT) and aspartate transaminase (AST) activities as well as the liver triglyceride (TG) and free fatty acid (FFA) levels were tested using reagent kits. Inflammatory cytokines, including Tnf-α, Il-ß and Il-6, and fibrosis genes, including Acta2, Col1a1, Col1a2 and Tgf-ß were examined by real-time quantitative PCR (RT-qPCR). Hematoxylin-eosin (H&E), Oil Red O, Masson's and Sirius Red staining were used to observe liver changes. The plasma and liver ceramide levels were analyzed using HPLC-QQQ-MS/MS. The expression of serine palmitoyl-CoA transferase (Spt), ceramide synthase 6 (Cers6), dihydroceramide desaturase 1 (Des1), glucosylceramide synthase (Gcs), and ceramide kinase (Cerk) mRNA was assayed by RT-qPCR, while the protein expression of CerS6, DES1, GCS, CerK, and casein kinase 2α (CK2α) was tested by western blotting (WB). CerS6 degradation was evaluated using a cycloheximide (CHX) assay in vitro. RESULTS: The liver index decreased by 20% in DHG groups and the serum ALT and AST decreased by approximately 50% and 30%, respectively in the DHG-H group. The liver Oil Red O staining, TG, and FFA changes showed that DHG reduced hepatic lipid accumulation by approximately 30% in NASH rats. H&E, Masson's and Sirius Red staining and the mRNA levels of Tnf-α, Il-ß, Il-6, Acta2, Col1a1, Col1a2 and Tgf-ß revealed that DHG alleviated liver inflammation and fibrosis in NASH rats. The ceramide (Cer 16:0), and hexosylceramide (HexCer 16:0, HexCer 18:0, HexCer 22:0, HexCer 24:0 and HexCer 24:1) levels decreased by approximately 17-56% in the plasma of the DHG-M and H rats. The Cer 16:0 content in the liver decreased by 20%, 50%, and 70% with the DHG-L, M, and H treatments; additionally, the dhCer 16:0, Cer 18:0, HexCer 18:0, HexCer 20:0 Cer 22:0-1P, Cer 24:0-1p, Cer 24:1-1p, and Cer 26:1-1p levels decreased in the DHG groups. The mRNA and protein expression levels of DES1, GCS, Cerk, CerS6, and CHX assay indicated that DHG decreased the mRNA and protein expression levels of CerK and reduced CerS6 protein expression by promoting its degradation. Additionally, DHG attenuated the protein expression of CK2α which could increase CerS6 enzymatic activity by phosphorylating its C-terminal region. CONCLUSION: DHG ameliorated the levels of liver FFA and TG and inflammation and fibrosis in MCD-induced rats, which were associated with decreasing ceramide species in the plasma and liver by reducing the expression levels of CerS6 and CerK.


Assuntos
Hepatopatia Gordurosa não Alcoólica , Animais , Anti-Inflamatórios/farmacologia , Ceramidas/metabolismo , Ceramidas/farmacologia , Ceramidas/uso terapêutico , Fibrose , Interleucina-6/metabolismo , Fígado , Cirrose Hepática/metabolismo , Metionina/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/metabolismo , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Esfingosina N-Aciltransferase/metabolismo , Espectrometria de Massas em Tandem , Fator de Crescimento Transformador beta/metabolismo , Triglicerídeos , Fator de Necrose Tumoral alfa/metabolismo
10.
Food Chem ; 393: 133322, 2022 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-35661467

RESUMO

It is necessary to obtain more recent data on the prevalence and co-occurrence of mycotoxins in feed and food to minimize risks. This study examined the recent presence, co-occurrence, and correlation patterns of six major ergot alkaloids (EAs; i.e. ergocornine, ergocristine, ergocryptine, ergometrine, ergosine, and ergotamine) in cool-season adapted barley (n = 57) and wheat (n = 80) submitted by livestock producers and industries for testing ergot alkaloids/mycotoxins by liquid chromatography - tandem mass spectrometry method. Overall, 91% industry-submitted barley samples and 84% industry-submitted wheat samples tested positive for at least one ergot alkaloid and 33% industry-submitted barley and 38% industry-submitted wheat samples were found to be co-contaminated with all six major EAs. The content of total EAs in 9 industry-submitted barley (16%) and 18 industry-submitted wheat (23%) samples exceeded the recommended maximum allowable level for lactating or pregnant animals (250 ppb). All the barley and wheat samples that contained detectable ergosine were found to co-occur with other EAs. Overall, the content of individual EAs was positively correlated with each other and strong correlations (r > 0.8, P < 0.01) were detected between the content of individual EAs and total EAs. These results implied that the industry and producers submitted cool-season adapted barley and wheat samples contaminated with a single EA is likely to contain high levels of other major EAs. The patterns of individual EAs in this study were distinct from previous studies that focus on samples from European countries. Ergocristine was remained as the predominant EAs in the industry-submitted cool-season adapted barley and wheat samples at levels up to 9438.8 and 12416.2 ppb, respectively. While the mean contents of ergosine were the lowest (68.5 and 50.6 ppb for the industry-submitted cool-season adapted barley and wheat samples, respectively). The high prevalence and co-occurrence of EAs indicated that ergot contamination is still posing a significant threat to food and feed industry and more research is expected to reduce the contamination level and explore the toxicological significance of various co-occurrence profiles. A full scale of investigation for all barley and wheat samples is needed to obtain a full picture of mycotoxin contamination.


Assuntos
Alcaloides de Claviceps , Hordeum , Micotoxinas , Animais , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Grão Comestível/química , Alcaloides de Claviceps/análise , Contaminação de Alimentos/análise , Lactação , Micotoxinas/análise , Prevalência , Estações do Ano , Análise Espectral , Espectrometria de Massas em Tandem/métodos , Triticum
11.
Crit Rev Eukaryot Gene Expr ; 32(4): 1-9, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35695660

RESUMO

Purpose - The present study aimed to identify differently expressed peptides involved in BC as potential biomarkers. Experimental Design - The serum proteomic profiling of 128 serum samples from 64 BC patients and 64 healthy controls (HC), using magnetic beads based immobilized metal ion affinity chromatography (MB-IMAC-Cu) separation followed by MALDI-TOF MS. ClinProTools software identified a number of distinct markers. Then, we performed liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) to identify the candidate serum biomarker based on serum proteomics analysis. Finally, enzyme-linked immunosorbent assays (ELISAs) were used to verify the expression of the candidate serum biomarker in BC patients. Results - BC patients could be identified with sensitivity of 87.32% and specificity of 89.46%. Of 41 m/z peaks that differed between BC and HC, six peaks were significantly different between BC and HC (P < 0.01, fold change > 1.5), with peak 1 upregulated and peaks 2-6 downregulated in the BC group. The upregulated peak 1 (m/z: 6638.63) is identified as a region of apolipoprotein C1 (APOC1), and validation showed that APOC1 expression increased from healthy controls to those with FA as well as mastopathy, and finally BC patients. Conclusions and Clinical Relevance - The present study indicates that APOC1 could serve as a candidate serum diagnostic biomarker for BC.


Assuntos
Apolipoproteína C-I/sangue , Neoplasias da Mama , Proteômica , Biomarcadores , Biomarcadores Tumorais , Neoplasias da Mama/diagnóstico , Feminino , Humanos , Proteômica/métodos , Espectrometria de Massas em Tandem
12.
J Ethnopharmacol ; 295: 115439, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-35667581

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Viscum cruciatum Sieb is a well-known medicinal plant in Jordan containing various secondary metabolites. It has traditionally been used to treat many ailments, most notably cancer. However, there is a significant gap between scientific research and its value in traditional medicine. AIM OF THE WORK: To evaluate the antiproliferative activity of different V. cruciatum extracts against MCF-7 breast cancer cell lines and recognize the affected cell cycle phase. Besides, identifying the bioactive components present in the active extract using LC/MS technique. Also, to determine the possible mechanism of action by in silico and in-vitro study. MATERIALS AND METHODS: V. cruciatum was extracted using solvents with increasing polarity. The antiproliferative effects of the extracts against MCF-7 cell lines were evaluated using SRB assay. Further, flow cytometry was used to identify the inhibited phase of the cell cycle, while LC/MS-MS technique was used to analyze the chemical composition of the most active extract. After that, the putative mechanism of action was investigated through in-silico docking, molecular dynamic simulation for compounds with the highest docking scores, and Western blot analysis of cyclin-dependent kinases (CDK2/4/6). RESULTS: The chloroform/methanol 90/10 (ChMe) extract showed the most potent antiproliferative effect against MCF-7 cells (IC50 = 23.8 µg/mL), and cell cycle arrest at the G0/G1phase. Furthermore, LC-MS/MS analysis revealed the presence of several polyphenolics belonging to the flavonoids and phenolic acids classes. Additionally, quercetin-4'-glucoside, 3, 5, 7-trihydroxy-4'-methoxy flavone, and hesperetin-7-O-neohesperidoside demonstrated the highest docking binding scores and stable complexes against CDK2 and CDK4/6. Moreover, RMSD (root-mean-square deviation), RMSF (root-mean-square fluctuation), Rg (radius of gyration), and energy analysis during molecular dynamic simulation indicated the stable binding of the studied complexes. These results were supported by Western blot analysis, which revealed the downregulation of CDK2, CDK4, and CDK6 protein expression in MCF-7 cell lines. CONCLUSION: These findings emphasized the potential breast anticancer activity of the V. cruciatum ChMe extract by arresting the G0/G1 phase of the cell cycle, which could be related to its flavonoid content. Moreover, the results provided experimental support for the traditional anticancer activity of V. cruciatum, and its ChMe extract might be a source of chemoprotective or chemotherapeutic isolates.


Assuntos
Antineoplásicos Fitogênicos , Viscum , Antineoplásicos Fitogênicos/uso terapêutico , Apoptose , Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , Cromatografia Líquida , Flavonoides/farmacologia , Pontos de Checagem da Fase G1 do Ciclo Celular , Humanos , Células MCF-7 , Extratos Vegetais/uso terapêutico , Espectrometria de Massas em Tandem
13.
Biomed Pharmacother ; 152: 113222, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35671581

RESUMO

Empagliflozin (EMPA) is the first sodium-glucose co-transporter 2 inhibitor to significantly reduce cardiovascular and kidney complications in type 2 diabetes mellitus. Given this, we speculate that EMPA may have the potential to intervene in diabetic retinopathy (DR), which is another diabetes-specific microvascular complication. Db/db mice were treated with EMPA for different periods to observe the retinas and related mechanisms. EMPA effectively balanced body weight and blood glucose levels, mitigated ocular edema and microaneurysm in db/db mice. EMPA significantly inhibited oxidative stress, apoptosis and recovered tight junction in diabetic retinas. MS/MS analyses showed that EMPA suppressed aberrant branched-chain amino acid (BCAAs) accumulation in db/db retinas, which led to the inhibition of the mammalian target of rapamycin activation, downregulation of inflammation, and angiogenic factors, including TNF-ɑ, IL-6, VCAM-1, and VEGF induced by diabetes. Furthermore, branched-chain α-keto acids (BCKAs), which are catabolites of BCAAs, were increased in diabetic retinas and decreased with EMPA application. Moreover, branched-chain ketoacid dehydrogenase kinase (BCKDK) was enhanced, BCKDHA and BCKDHB were decreased in diabetic retinas. This could be reversed by EMPA treatment, thus promoting BCAAs catabolism to decrease BCAAs and BCKAs accumulation in diabetic retinas. The high levels of BCAAs in the plasma and enhanced L-type amino acid transporter 1 (LAT1) were responsible for the high levels of BCAAs in diabetic retinas, which could be inhibited by EMPA. Overall, EMPA could ameliorate DR manifestations. The normalization of BCAAs catabolism and intake may play a role in this process. This study supports EMPA as a protective drug against DR.


Assuntos
Diabetes Mellitus Tipo 2 , Retinopatia Diabética , Inibidores do Transportador 2 de Sódio-Glicose , Aminoácidos de Cadeia Ramificada , Animais , Compostos Benzidrílicos/farmacologia , Compostos Benzidrílicos/uso terapêutico , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/tratamento farmacológico , Retinopatia Diabética/tratamento farmacológico , Glucosídeos , Mamíferos , Camundongos , Inibidores do Transportador 2 de Sódio-Glicose/farmacologia , Inibidores do Transportador 2 de Sódio-Glicose/uso terapêutico , Espectrometria de Massas em Tandem
14.
Anal Chem ; 94(24): 8580-8587, 2022 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-35678765

RESUMO

Additive manufacturing (3D printing) has greatly revolutionized the way researchers approach certain technical challenges. Despite its outstanding print quality and resolution, stereolithography (SLA) printing is cost-effective and relatively accessible. However, applications involving mass spectrometry (MS) are few due to residual oligomers and additives leaching from SLA-printed devices that interfere with MS analyses. We identified the crosslinking agent urethane dimethacrylate as the main contaminant derived from SLA prints. A stringent washing and post-curing protocol mitigated sample contamination and rendered SLA prints suitable for MS hyphenation. Thereafter, SLA printing was used to produce 360 µm I.D. microcolumn chips with excellent structural properties. By packing the column with polystyrene microspheres and covalently immobilizing pepsin, an exceptionally effective microscale immobilized enzyme reactor (µIMER) was created. Implemented in an online liquid chromatography-MS/MS setup, the protease microcolumn enabled reproducible protein digestion and peptide mapping with 100% sequence coverage obtained for three different recombinant proteins. Additionally, when assessing the µIMER digestion efficiency for complex proteome samples, it delivered a 144-fold faster and significantly more efficient protein digestion compared to 24 h for bulk digestion. The 3D-printed µIMER withstands remarkably high pressures above 130 bar and retains its activity for several weeks. This versatile platform will enable researchers to produce tailored polymer-based enzyme reactors for various applications in analytical chemistry and beyond.


Assuntos
Enzimas Imobilizadas , Espectrometria de Massas em Tandem , Cromatografia Líquida , Enzimas Imobilizadas/química , Mapeamento de Peptídeos/métodos , Impressão Tridimensional
15.
Anal Chem ; 94(24): 8625-8632, 2022 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-35679579

RESUMO

Polysorbates are nonionic surfactants that have been widely used in biotherapeutic formulations to prevent protein aggregation and denaturation. However, polysorbates are subject to degradation after prolonged storage if certain lipases are present in the biotherapeutic product. Because the degradation of polysorbates compromises the shelf life of biotherapeutics and leads to the formation of undesirable products such as protein aggregates and subvisible particles, it is important to identify the active enzymes that catalyze polysorbate hydrolysis. In this study, we developed a novel fluorophosphonate activity-based protein profiling (ABPP) probe (termed the REGN probe), which mimics the structure of polysorbate and targets lipases catalyzing polysorbate degradation. We demonstrated that the REGN probe could enrich certain lipases from Chinese hamster ovary (CHO) cell lysate by more than 100-fold compared with direct tryptic digestion. Furthermore, we found that the REGN probe had higher lipase enrichment efficiency than commercially available ABPP probes including fluorophosphonate-biotin (FP-biotin) and FP-desthiobiotin. Remarkably, the REGN probe can enrich several lipases that cannot be labeled by commercial probes, such as lysosomal acid lipase and cytosolic phospholipase A2. Additionally, we showed that lipases with abundances as low as 0.08 ppm in drug substances were detected by the REGN probe enrichment and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Collectively, we have developed a novel ABPP probe with higher enrichment efficiency and broader coverage for lipases compared with commercial probes, and this probe can be used to detect the trace level of lipases in biotherapeutic products and to facilitate their development and manufacturing.


Assuntos
Polissorbatos , Espectrometria de Massas em Tandem , Animais , Células CHO , Cromatografia Líquida , Cricetinae , Cricetulus , Lipase , Polissorbatos/química , Tensoativos/química
16.
Sci Total Environ ; 839: 156260, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-35644406

RESUMO

The miniaturization of a full workflow for identification and monitoring of contaminants of emerging concern (CECs) is presented. Firstly, successful development of a low-cost small 3D-printed passive sampler device (3D-PSD), based on a two-piece methacrylate housing that held up to five separate 9 mm disk sorbents, is discussed. Secondly, a highly sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method reduced the need for large scale in-laboratory apparatus, solvent, reagents and reference material quantities for in-laboratory passive sampler device (PSD) calibration and extraction. Using hydrophilic-lipophilic balanced sorbents, sampling rates (Rs) were determined after a low 50 ng L-1 exposure over seven days for 39 pesticides, pharmaceuticals, drug metabolites and illicit drugs over the range 0.3 to 12.3 mL day-1. The high sensitivity LC-MS/MS method enabled rapid analysis of river water using only 10 µL of directly injected sample filtrate to measure occurrence of 164 CECs and sources along 19 sites on the River Wandle, (London, UK). The new 3D-PSD was then field-tested over seven days at the site with the highest number and concentration of CECs, which was down-river from a wastewater treatment plant. Almost double the number of CECs were identified in 3D-PSD extracts across sites in comparison to water samples (80 versus 42 CECs, respectively). Time-weighted average CEC concentrations ranged from 8.2 to 845 ng L-1, which were generally comparable to measured concentrations in grab samples. Lastly, high resolution mass spectrometry-based suspect screening of 3D-PSD extracts enabled 113 additional compounds to be tentatively identified via library matching, many of which are currently or are under consideration for the EU Watch List. This miniaturized workflow represents a new, cost-effective, and more practically efficient means to perform passive sampling chemical monitoring at a large scale. SYNOPSIS: Miniaturized, low cost, multi-disk passive samplers enabled more efficient multi-residue chemical contaminant characterization, potentially for large-scale monitoring programs.


Assuntos
Monitoramento Ambiental , Poluentes Químicos da Água , Cromatografia Líquida , Monitoramento Ambiental/métodos , Espectrometria de Massas em Tandem , Águas Residuárias/análise , Água/análise , Poluentes Químicos da Água/análise , Fluxo de Trabalho
17.
Int J Cardiol ; 362: 118-125, 2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-35662556

RESUMO

BACKGROUND: Although epicardial adipose tissue (EAT) is known to be a major contributor to the pathogenesis of heart failure with preserved ejection fraction (HFpEF), the underlying mechanisms remain incompletely understood. This study aimed to compare the proteomic profiles of EAT from HFpEF patients and patients without HF (non-HF) and to explore candidate molecules characteristic of EAT in HFpEF. METHODS: EAT samples were collected from patients who underwent cardiac surgery. Proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and protein-protein interaction network analysis were conducted. The gene expression of one significant differentially expressed protein was examined by quantitative reverse transcription polymerase chain reaction. RESULTS: A total of 2416 proteins were detected by LC-MS/MS experiments, and expression levels were quantified for 2349 proteins. Among them, 96 proteins (including 71 upregulated proteins and 25 downregulated proteins) were significantly differentially expressed between the HFpEF (n = 5) and non-HF groups (n = 5). GO enrichment and KEGG pathway analyses revealed that these differentially expressed proteins were predominantly involved in HFpEF-related processes, including lipid metabolic disorder, inflammation, and mitochondrial dysfunction. CONCLUSIONS: The results of this comprehensive analysis of the EAT proteome in HFpEF patients offer new insights into the pathogenesis of HFpEF and potential molecular targets in EAT.


Assuntos
Insuficiência Cardíaca , Tecido Adiposo/metabolismo , Cromatografia Líquida , Humanos , Proteômica , Volume Sistólico , Espectrometria de Massas em Tandem
18.
J Agric Food Chem ; 70(24): 7500-7514, 2022 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-35674110

RESUMO

Sensory-guided fractionation by means of ultrafiltration and gel permeation chromatography followed by high-performance liquid chromatography, synthesis, liquid chromatography-tandem mass spectrometry (LC-MS/MS) quantitation, and taste re-engineering experiments revealed taste-active and taste-enhancing compounds contributing to the umami, mouthful and complex taste profile of the fermented Korean dish, kimchi. Besides basic taste-active compounds, in particular, various N-acylated amino acids deriving from succinic acid and lactic acid imparted taste-modulating properties in food matrices. Taste threshold concentrations were determined to evaluate intrinsic and modulating effects. Quantitation of N-acylated amino acids in kimchi following synthesis revealed the presence of numerous derivatives showing taste-active properties. Sensory evaluation including recombination and partial addition experiments highlighted that both the N-lactoyl- and the N-succinoyl amino acid derivatives contribute to increasing the fullness, volume, and complexity of food matrices, whereas the latter directly contributes to the overall taste of kimchi in natural concentrations.


Assuntos
Paladar , Aminoácidos/química , Cromatografia Líquida , República da Coreia , Espectrometria de Massas em Tandem
19.
J Agric Food Chem ; 70(24): 7594-7606, 2022 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-35674269

RESUMO

Bakery products containing poppy seeds are increasingly being commercialized. These seeds may be contaminated with latex from the Papaver somniferum L. plant rich in opium alkaloids (OAs). Therefore, health authorities demand the development of analytical methods to control them. In this study, an efficient and simple method was developed and validated for the first time to analyze six OAs in bakery products by high-performance liquid chromatography-tandem mass spectrometry. For this purpose, a solid-liquid extraction was optimized, and then a magnetic material [magnetite surface-modified with Fe(III) terephthalate, denoted as Fe3O4@TPA-Fe] was used for a fast magnetic solid-phase extraction. The method has been validated with adequate recoveries (70-110%) and relative standard deviations (<20%) and without matrix effects. Nine bakery samples (five breadsticks and four sliced bread) were analyzed; breadsticks showed low amounts of OAs, but two sliced bread showed higher amounts of OAs than the new amount (1.5 mg/kg) set by the Commission Regulation (EU) 2021/2142.


Assuntos
Alcaloides Opiáceos , Papaver , Cromatografia Líquida de Alta Pressão/métodos , Compostos Férricos , Fenômenos Magnéticos , Ópio/análise , Papaver/química , Sementes/química , Extração em Fase Sólida , Espectrometria de Massas em Tandem/métodos
20.
Environ Pollut ; 307: 119577, 2022 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-35688393

RESUMO

Inhalation exposure to fine particulate matter (PM2.5) represents a global concern due to the adverse effects in human health. In the last years, scientific community has been adopted the assessment of the PM2.5-bound pollutant fraction that could be released (bioaccessible fraction) in simulated lung fluids (SLFs) to achieve a better understanding of PM risk assessment and toxicological studies. Thus, bioaccessibility of 49 organic pollutants, including 18 polycyclic aromatic hydrocarbons (PAHs), 12 phthalate esters (PAEs), 11 organophosphorus flame retardants (OPFRs), 6 synthetic musk compounds (SMCs) and 2 bisphenols in PM2.5 samples was evaluated. The proposed method consists of a physiologically based extraction test (PBET) by using artificial lysosomal fluid (ALF) to obtain bioaccessible fractions, followed by a vortex-assisted liquid-liquid microextraction (VALLME) and a final analysis by programmed temperature vaporization-gas chromatography-tandem mass spectrometry (PTV-GC-MS/MS). The highest inhalation bioaccessibility ratio was found for bisphenol A (BPA) with an average of 83%, followed by OPFRs, PAEs and PAHs (with average bioaccessibilities of 68%, 41% and 34%, respectively). Correlations between PM2.5 composition (major ions, trace metals, equivalent black carbon (eBC) and UV-absorbing particulate matter (UVPM)) and bioaccessibility ratios were also assessed. Principal Component Analysis (PCA) suggested that PAHs, PAES and OPFRs bioaccessibility ratios could be positively correlated with PM2.5 carbonaceous content. Furthermore, both inverse and positive correlations on PAHs, PAEs and OPFRs bioaccessibilites could be accounted for some major ions and metal (oid)s associated to PM2.5, whereas no correlations comprising considered PM2.5 major ions and metal (oid)s contents and BPA bioaccessibility was observed. In addition, health risk assessment of target PM2.5-associated PAHs via inhalation was assessed in the study area considering both total and bioaccessible concentrations, being averaged human health risks within the safe carcinogenic and non-carcinogenic levels.


Assuntos
Poluentes Atmosféricos , Poluentes Ambientais , Retardadores de Chama , Hidrocarbonetos Policíclicos Aromáticos , Poluentes Atmosféricos/análise , Monitoramento Ambiental , Poluentes Ambientais/análise , Retardadores de Chama/análise , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Material Particulado/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Medição de Risco , Espectrometria de Massas em Tandem
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...