RESUMO
Salinity is one of the most serious threats to sustainable agriculture. The Salt Overly Sensitive (SOS) signaling pathway plays an important role in salinity tolerance in plants, and the SOS2 gene plays a critical role in this pathway. Mulberry not only has important economic value but also is an important ecological tree species; however, the roles of the SOS2 gene associated with salt stress have not been reported in mulberry. To gain insight into the response of mulberry to salt stress, SOS2 (designated MulSOS2) was cloned from mulberry (Morus atropurpurea Roxb), and sequence analysis of the amino acids of MulSOS2 showed that it shares some conserved domains with its homologs from other plant species. Our data showed that the MulSOS2 gene was expressed at different levels in different tissues of mulberry, and its expression was induced substantially not only by NaCl but also by ABA. In addition, MulSOS2 was exogenously expressed in Arabidopsis, and the results showed that under salt stress, transgenic MulSOS2 plants accumulated more proline and less malondialdehyde than the wild-type plants and exhibited increased tolerance to salt stress. Moreover, the MulSOS2 gene was transiently overexpressed in mulberry leaves and stably overexpressed in the hairy roots, and similar results were obtained for resistance to salt stress in transgenic mulberry plants. Taken together, the results of this study are helpful to further explore the function of the MulSOS2 gene, which provides a valuable gene for the genetic breeding of salt tolerance in mulberry.
Assuntos
Arabidopsis , Morus , Tolerância ao Sal/genética , Morus/genética , Melhoramento Vegetal , Estresse Salino , Agricultura , Plantas Geneticamente ModificadasRESUMO
Objective This consensus aims to provide evidence-based recommendations on common questions in the diagnosis and treatment of acute respiratory failure (ARF) for critically ill cancer patients.Methods We developed six clinical questions using the PICO (Population, Intervention, Comparison, and Outcome) principle in diagnosis and treatment for critical ill cancer patients with ARF. Based on literature searching and meta-analyses, recommendations were devised. The GRADE (Grading of Recommendation Assessment, Development and Evaluation) method was applied to each question to reach consensus in the expert panel. Results The panel makes strong recommendations in favor of (1) metagenomic next-generation sequencing (mNGS) tests may aid clinicians in rapid diagnosis in critically ill cancer patients suspected of pulmonary infections; (2) extracorporeal membrane oxygenation (ECMO) therapy should not be used as a routine rescue therapy for acute respiratory distress syndrome in critically ill cancer patients but may benefit highly selected patients after multi-disciplinary consultations; (3) cancer patients who have received immune checkpoint inhibitor therapy have an increased incidence of pneumonitis compared with standard chemotherapy; (4) critically ill cancer patients who are on invasive mechanical ventilation and estimated to be extubated after 14 days may benefit from early tracheotomy; and (5) high-flow nasal oxygen and noninvasive ventilation therapy can be used as a first-line oxygen strategy for critically ill cancer patients with ARFs. A weak recommendation is: (6) for critically ill cancer patients with ARF caused by tumor compression, urgent chemotherapy may be considered as a rescue therapy only in patients determined to be potentially sensitive to the anticancer therapy after multidisciplinary consultations. Conclusions The recommendations based on the available evidence can guide diagnosis and treatment in critically ill cancer patients with acute respiratory failure and improve outcomes.
Assuntos
Neoplasias , Pneumonia , Síndrome do Desconforto Respiratório , Insuficiência Respiratória , Humanos , Consenso , Estado Terminal/terapia , Neoplasias/complicações , Neoplasias/terapia , Oxigênio , Síndrome do Desconforto Respiratório/terapia , Síndrome do Desconforto Respiratório/tratamento farmacológico , Insuficiência Respiratória/etiologia , Insuficiência Respiratória/terapiaRESUMO
Ru-based catalysis results in highly unsaturated fatty acid (HUFA) ethyl esters (EE) deuterated to various extents. The products carry 2H (D) mainly at their bis-allylic positions, where they are resistant to autoxidation compared to natural HUFA and are promising as neurological and retinal drugs. We characterized the extent of deuteration at each allylic position of docosa-4,7,10,13,16,19-hexaenoic acid deuterated to completion at bis-allylic and allylic positions (D-DHA) by two-dimensional (2D) and high-field (600 and 950 MHz) NMR. In separate experiments, the kinetics of docosahexaenoic acid (DHA) EE deuteration was evaluated using Paternò-Büchi (PB) reaction tandem mass spectrometry (MS/MS) analysis, enabling deuteration to be quantitatively characterized for isotopologues (D0-D14 DHA) at each internal allylic position. NMR analysis shows that the net deuteration of the isotopologue mixture is about 94% at the bis-allylic positions, and less than 1% remained as the protiated -CH2-. MS analysis shows that deuteration kinetics follow an increasing curve at bis-allylic positions with higher rate for internal bis-allylic positions. Percent D of bis-allylic positions increases linearly from D1 to D9 in which all internal bis-allylic positions (C9, C12, C15) deuterate uniformly and more rapidly than external bis-allylic positions (C6, C18). The mono-allylic positions near the methyl end (C21) show a steep increase of D only after the D10 isotopologue has been deuterated to >90%, while the mono-allylic position near the carboxyl position, C3, deuterates last and least. These data establish detailed methods for the characterization of Ru-catalyzed deuteration of HUFA as well as the phenomenological reaction kinetics as net product is formed.
Assuntos
Ácidos Docosa-Hexaenoicos , Ácidos Graxos , Catálise , Ácidos Graxos Insaturados , Imidazóis , Sulfonamidas , Espectrometria de Massas em Tandem , TiofenosRESUMO
No general method currently is available for the quantitative determination of deuterium (D) at C positions along a hydrocarbon chain. Bis-allylic deuterated highly unsaturated fatty acids (D-HUFA) are a novel class of drugs stabilized against H-abstraction-mediated oxidation by deuteration at the most labile positions. Ru-based catalytic deuteration overcomes the limited scale of bis-allylic D-HUFA production by total organic synthesis; however, it produces a complex mixture of bis-allylic D isotopologues and isotopomers, requiring detailed sequencing for characterization. We report here adaptation and application of the Paternó-Büchi (PB) reaction of 2-acetylpyridine to a series of D-HUFA with analysis by shotgun lipidomics to determine position-specific quantitative D abundances. Sodiated PBD-HUFA result in diagnostic ions of high abundance upon collision-induced dissociation (CID) activation, enabling sensitive differentiation and quantification of D fraction at each bis- and mono-allylic position for each isotopologue. Catalytically deuterated isotopologues D5-7 linolenic acid (D5-7 LnA), D6-8 arachidonic acid (D6-8 ARA), D7-9 eicosapentaenoic acid (D7-9 EPA), and D9-11 docosahexaenoic acid (D9-11 DHA) incorporate 80-98, 95-100, 81-100, and 83-100% D at their bis-allylic positions, respectively. D-HUFA isotopologues having D number greater than or equal to bis-allylic sites (e.g., D10-DHA or D11-DHA) deuterated >95% at bis-allylic positions, except for D-LnA. The mono-allylic position near the methyl end deuterates to a much greater extent than the mono-allylic position near the carboxyl end, and both positions deuterate only when bis-allylic D is near-saturated. This method enables rapid, accurate characterization of position and isotopomer-specific D composition and enables sequencing along the chain.
Assuntos
Ácidos Graxos Insaturados , Ácidos Graxos , Deutério , Ácidos Docosa-Hexaenoicos , Hidrocarbonetos , OxirreduçãoRESUMO
Salvia miltiorrhiza is a renowned model medicinal plant species for which 15 SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) family genes have been identified; however, the specific functions of SmSPLs have not been well characterized as of yet. For this study, the expression patterns of SmSPL6 were determined through its responses to treatments of exogenous hormones, including indole acetic acid (IAA), gibberellic acid (GA3), methyl jasmonic acid (MeJA), and abscisic acid (ABA). To characterize its functionality, we obtained SmSPL6-ovexpressed transgenic S. miltiorrhiza plants and found that overexpressed SmSPL6 promoted the accumulation of phenolic acids and repressed the biosynthesis of anthocyanin. Meanwhile, the root lengths of the SmSPL6-overexpressed lines were significantly longer than the control; however, both the fresh weights and lateral root numbers decreased. Further investigations indicated that SmSPL6 regulated the biosynthesis of phenolic acid by directly binding to the promoter regions of the enzyme genes Sm4CL9 and SmCYP98A14 and activated their expression. We concluded that SmSPL6 regulates not only the biosynthesis of phenolic acids, but also the development of roots in S. miltiorrhiza.
Assuntos
Regulação da Expressão Gênica de Plantas , Hidroxibenzoatos/metabolismo , Organogênese Vegetal , Proteínas de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Salvia miltiorrhiza/crescimento & desenvolvimento , Proteínas de Plantas/genética , Raízes de Plantas/metabolismo , Salvia miltiorrhiza/metabolismoRESUMO
Pine nuts and other edible gymnosperm seeds contain unusual, bioactive polymethylene-interrupted polyunsaturated fatty acids (PMI-PUFAs), a subset of nonmethylene-interrupted PUFA with (-CH2-)n≥2 intervening between double bonds. Conventional methods for structure elucidation of PMI-PUFAs require special derivatization risking rearrangement artifacts. Here we introduce a facile solvent-mediated (SM) covalent adduct chemical ionization (CACI) system modified with a triple quadrupole MS, which distinguishes PMI-PUFAs from their analogues in direct methyl ester form. The prominent Δ5 desaturated PMI-PUFAs exhibit characteristic fragmentation at C6-7 to yield ω diagnostic ions and share their fragmentation pattern with normal methylene interrupted PUFAs for the α diagnostic ion. H⢠transfer upon CID dissociation of PMI-PUFAs was found to be dependent on the relative position of isolated lone double bonds and cleavage points. Ginkgo and five species of pine nuts were characterized for their unique Δ5 fatty acid profile, without the need for chemical standards.
Assuntos
Ácidos Graxos Insaturados/análise , Estrutura Molecular , Pinus/química , Sementes/química , Solventes/química , Espectrometria de Massas em TandemRESUMO
RATIONALE: Quantitative analysis of triacylglycerols (TAG) is impeded by a lack of standards and the huge number of potential TAG molecular species that may be present due to the combinatorial nature of glycerolipids. Randomization of acyl groups yields TAG mixtures with profiles predictable from fatty acid profiles; however, their use as calibration mixtures has been limited. METHODS: We introduce here the principle of fatty acid isotopic sentinels that are quantitatively added prior to randomization to enable verification that randomization is complete, and that can be used as internal standards. A mixture of two isotopically labeled fatty acid methyl esters (FAME) is prepared at a ratio of 2:1 and the exact ratio is carefully measured by gas chromatography flame ionization detection (GC-FID) and randomized covalently into the acyl groups of TAG mixtures. RESULTS: Reaction with catalytic amounts of NaOCH3 yields complete randomization, such that the product FAME and TAG have the same fatty acid profile. TAG mixture analysis reveals that the isotopic sentinels have been covalently incorporated into the TAG molecular species at <1% deviation from the expected proportions, thus verifying randomization within experimental error. CONCLUSIONS: The sentinel principle demonstrated here as covalently incorporated internal standards verifies that randomization chemistry went to completion. It applies in general to use of combinatorial chemistry for quantitative standards.
RESUMO
Monounsaturated normal fatty acids (n-MUFA) and saturated branched chain fatty acids (BCFA) are structurally characterized by separate tandem mass spectrometry methods for double bond localization and for chain branching in their respective fatty acid methyl ester (FAME) derivatives; however, these methods have never been applied to branched monounsaturated FAME. Here, we report application of electron ionization (EI)-MS/MS and solvent-mediated covalent adduct chemical ionization (CACI)-MS/MS of monounsaturated BCFA methyl esters (MUBCFAME) of a chain length of 15-20 carbons. A novel system was used to implement CI with low vapor pressure reagents in a tabletop triple quadrupole MS. Anteiso-MUBCFA EI-MS/MS of the molecular ion (M) yields a characteristic diagnostic ion [M-29]+. iso-MUBCFA can be distinguished from n-MUFA by an ion intensity ratio of [M-32]+/[M-43]+, with iso-MUBCFA yielding a ratio greater than 1.7, while n-MUFA yields a ratio less than 1.0. Chain branching at the iso and anteiso positions, terminal isopropyl and sec-butyl, respectively, do not alter CACI-MS/MS diagnostic ions compared to normal BCFA, enabling double bond positions of MUBCFA to be determined with the analogous α and ω diagnostic ions from cleavage on both sides of the erstwhile double bond. Taken together, this straightforward FAME-based technique via combination of EI-MS/MS and CACI-MS/MS enables fundamental structural identification of MUBCFA without standards.
Assuntos
Ésteres/química , Ácidos Graxos Monoinsaturados/química , Espectrometria de Massas em Tandem/métodos , Elétrons , Íons/química , Estrutura Molecular , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
The role of lipid metabolism in epithelial stem cell (SC) function and carcinogenesis is poorly understood. The transcription factor Runx1 is known to regulate proliferation in mouse epithelial hair follicle (HF) SCs in vivo and in several mouse and human epithelial cancers. We found a novel subset of in vivo Runx1 HFSC target genes related to lipid metabolism and demonstrated changes in distinct classes of lipids driven by Runx1. Inhibition of lipid-enzymes Scd1 and Soat1 activity synergistically reduces proliferation of mouse skin epithelial cells and of human skin and oral squamous cell carcinoma cultured lines. Varying Runx1 levels induces changes in skin monounsaturated fatty acids (e.g., oleate, a product of Scd1) as shown by our lipidome analysis. Furthermore, varying Runx1 levels, the inhibition of Scd1, or the addition of Scd1-product oleate, individually affects the plasma membrane organization (or fluidity) in mouse keratinocytes. These factors also affect the strength of signal transduction through the membranes for Wnt, a pathway that promotes epithelial (cancer) cell proliferation and HFSC activation. Our working model is that HFSC factor Runx1 modulates the fatty acid production, which affects membrane organization, facilitating signal transduction for rapid proliferation of normal and cancer epithelial cells. Stem Cells 2018;36:1603-1616.
Assuntos
Subunidade alfa 2 de Fator de Ligação ao Core/metabolismo , Células Epiteliais/metabolismo , Estearoil-CoA Dessaturase/metabolismo , Esterol O-Aciltransferase/metabolismo , Animais , Linhagem Celular Tumoral , Proliferação de Células/fisiologia , Subunidade alfa 2 de Fator de Ligação ao Core/biossíntese , Subunidade alfa 2 de Fator de Ligação ao Core/genética , Células Epiteliais/citologia , Humanos , Queratinócitos/citologia , Queratinócitos/metabolismo , Metabolismo dos Lipídeos/genética , Camundongos , Camundongos Knockout , Neoplasias Bucais/metabolismo , Neoplasias Bucais/patologia , Transdução de Sinais , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia , Carcinoma de Células Escamosas de Cabeça e Pescoço/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologia , Estearoil-CoA Dessaturase/genética , Células-Tronco/citologia , Células-Tronco/metabolismo , Esterol O-Aciltransferase/genética , TransfecçãoRESUMO
Normal odd-chain SFA (OCSFA), particularly tridecanoic acid (n-13 : 0), pentadecanoic acid (n-15 : 0) and heptadecanoic acid (n-17 : 0), are normal components of dairy products, beef and seafood. The ratio of n-15 : 0:n-17 : 0 in ruminant foods (dairy products and beef) is 2:1, while in seafood and human tissues it is 1:2, and their appearance in plasma is often used as a marker for ruminant fat intake. Human elongases encoded by elongation of very long-chain fatty acid (ELOVL)1, ELOVL3, ELOVL6 and ELOVL7 catalyse biosynthesis of the dominant even-chain SFA; however, there are no reports of elongase function on OCSFA. ELOVL transfected MCF7 cells were treated with n-13 : 0, n-15 : 0 or n-17 : 0 (80 µm) and products analysed. ELOVL6 catalysed elongation of n-13 : 0ân-15 : 0 and n-15 : 0ân-17 : 0; and ELOVL7 had modest activity toward n-15 : 0 (n-15 : 0ân-17 : 0). No elongation activity was detected for n-17 : 0ân-19 : 0. Our data expand ELOVL specificity to OCSFA, providing the first molecular evidence demonstrating ELOVL6 as the major elongase acting on OCSFA n-13 : 0 and n-15 : 0 fatty acids. Studies of food intake relying on OCSFA as a biomarker should consider endogenous human metabolism when relying on OCSFA ratios to indicate specific food intake.
RESUMO
BACKGROUND: Evidence suggests that excessive inflammation of the immature intestine may predispose premature infants to necrotizing enterocolitis (NEC). We investigated the anti-inflammatory effects of docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid (ARA) in human fetal and adult intestinal epithelial cells (IEC) in primary culture. METHODS: Human fetal IEC in culture were derived from a healthy fetal small intestine (H4) or resected small intestine of a neonate with NEC (NEC-IEC). Intestinal cell lines Caco2 and NCM460 in culture were used as models for mature IEC. IEC in culture were pretreated with 100 µmol/l palmitic acid (PAL), DHA, EPA, ARA, or ARA+DHA for 48 h and then stimulated with proinflammatory IL-1ß. RESULTS: DHA significantly attenuated IL-1ß induced proinflammatory IL-8 and IL-6 protein and mRNA in fetal H4, NEC-IEC, and mature Caco2, NCM460 IEC, compared to control and PAL treatment. DHA downregulated IL-1R1 (IL-1ß receptor) and NFk ß1 mRNA expression in fetal and adult IEC. ARA had potent anti-inflammatory effects with lower IL-8 and IL-6 (protein and mRNA) in fetal H4 but not in NEC-IEC or adult IEC. CONCLUSION: The present study provides evidence that DHA and ARA may have important anti-inflammatory functions for prevention of NEC in premature infants.
Assuntos
Anti-Inflamatórios/farmacologia , Ácido Araquidônico/farmacologia , Ácidos Docosa-Hexaenoicos/farmacologia , Enterocolite Necrosante/tratamento farmacológico , Células Epiteliais/efeitos dos fármacos , Íleo/efeitos dos fármacos , Mediadores da Inflamação/metabolismo , Interleucina-1beta/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Células CACO-2 , Citoproteção , Ácido Eicosapentaenoico/farmacologia , Enterocolite Necrosante/genética , Enterocolite Necrosante/metabolismo , Células Epiteliais/metabolismo , Regulação da Expressão Gênica , Humanos , Íleo/embriologia , Íleo/metabolismo , Recém-Nascido , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Mucosa Intestinal/embriologia , Mucosa Intestinal/metabolismo , Subunidade p50 de NF-kappa B/genética , Subunidade p50 de NF-kappa B/metabolismo , Ácido Palmítico/farmacologia , Receptores Tipo I de Interleucina-1/genética , Receptores Tipo I de Interleucina-1/metabolismoRESUMO
Branched-chain fatty acids (BCFA) are bioactive food components that constitute about 2 % of fatty acids in cows' milk fat. There are few systematic data available on the BCFA content of other foods to estimate dietary intakes. In the present study, we report BCFA distribution and content of fresh and processed foods representing the major foods in the American diet and estimate BCFA intake. BCFA are primarily components of dairy and ruminant food products, and are absent from chicken, pork and salmon. The mean BCFA intake of 500 mg/d was delivered primarily from dairy and beef food products; by comparison, average intake of the widely studied long-chain PUFA EPA and DHA has been estimated to be 100 mg/d. Common adjustments in the diet could double the daily intake of BCFA. The fermented foods sauerkraut and miso had appreciable fractions of BCFA, but, overall, they are low-fat foods providing very small amounts of BCFA in the diet, and other fermented foods did not contain BCFA as might have been expected from the influence of microbial exposure. These data support the quantitative importance of BCFA delivered primarily from dairy and beef food products and highlight the need for research into their effects on health.
Assuntos
Gorduras na Dieta/administração & dosagem , Ácidos Graxos/administração & dosagem , Análise de Alimentos , Adulto , Animais , Bovinos , Criança , Laticínios/análise , Gorduras na Dieta/análise , Gorduras na Dieta/metabolismo , Ácidos Graxos/análise , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Fermentação , Abastecimento de Alimentos , Alimentos em Conserva/análise , Alimentos em Conserva/microbiologia , Humanos , Isomerismo , Carne/análise , Produtos da Carne/análise , Estrutura Molecular , New York , Ruminantes , Alimentos de Soja/análise , Alimentos de Soja/microbiologia , Estados UnidosRESUMO
The biological functions of fatty acids and the lipids in which they are esterified are determined by their chain length, double bond position and geometry and other structural motifs such as the presence of methyl branches. Unusual isomeric features in fatty acids of human foods such as conjugated double bonds or chain branching found in dairy products, some seeds and nuts, and marine foods potentially have important effects on human health. Recent advancements in identifying fatty acids with unusual double bond positions and pinpointing the position of methyl branches have empowered the study of their biological functions. We present recent advances in fatty acid structural elucidation by mass spectrometry in comparison with the more traditional methods. The double bond position can be determined by purely instrumental methods, specifically solvent-mediated covalent adduct chemical ionization (SM-CACI) and ozone induced dissociation (OzID), with charge inversion methods showing promise. Prior derivatization using the Paternò-Büchi (PB) reaction to yield stable structures that, upon collisional activation, yield the double bond position has emerged. The chemical ionization (CI) based three ion monitoring (MRM) method has been developed to simultaneously identify and quantify low-level branched chain fatty acids (BCFAs), unattainable by electron ionization (EI) based methods. Accurate identification and quantification of unusual fatty acid isomers has led to research progress in the discovery of biomarkers for cancer, diabetes, nonalcoholic fatty liver disease (NAFLD) and atherosclerosis. Modulation of eicosanoids, weight loss and the health significance of BCFAs are also presented. This review clearly shows that the improvement of analytical capacity is critical in the study of fatty acid biological functions, and stronger coupling of the methods discussed here with fatty acid mechanistic research is promising in generating more refined outcomes.
Assuntos
Ácidos Graxos , Alimentos , Humanos , Ácidos Graxos/química , Isomerismo , Espectrometria de Massas/métodos , SolventesRESUMO
Fatty acids (FAs) containing polymethylene-interrupted (PMI) double bonds are a component of human foods; however, they present a significant analytical challenge for de novo identification. Covalent adduct chemical ionization and ozone-induced dissociation mass spectrometry (MS) methods enable unambiguous assignment of PMI-FA double bond positions, however, no method has been reported with electrospray ionization (ESI) platform using off-the-shelf systems. In the current work, we studied the Paternò-Büchi (PB) fragmentation patterns of PMI-FA and triacylglycerol (TG) by analyzing several known food sources. PB-MS/MS and MS3 enabled complete double bond location assignments, including the isolated double bond in PMI-FA and triacylglycerols. Sea urchin ("uni"), oyster, pine nut, and ginkgo nut were characterized for their signature PMI-FA, 20:2(5Z,11Z), 22:2(7Z, 15Z), 18:3(5Z,9Z,12Z), and 20:3(5Z,11Z,14Z), respectively. Quantitative analyses of the relative abundance of these PMI-FA led to results similar to reference methods. 18:3(5Z,9Z,12Z) was enriched at the sn-1/sn-3 position in pine nut major TG.
Assuntos
Ácidos Graxos , Espectrometria de Massas em Tandem , Humanos , Espectrometria de Massas em Tandem/métodos , Ácidos Graxos/química , Triglicerídeos/química , Alimentos , Espectrometria de Massas por Ionização por Electrospray/métodosRESUMO
Omega-3 long-chain polyunsaturated fatty acids (LCPUFA) play critical roles in human development and health. Their intake is often effectively estimated solely based on seafood consumption, though the high intake of terrestrial animal-based foods with minor amounts of LCPUFA may be significant. Covalent adduct chemical ionization (CACI) tandem mass spectrometry is one approach for de novo structural and quantitative analysis of minor unsaturated fatty acids (FA), for which standards are unavailable. Here, CACI-MS and MS/MS are used to identify and quantify minor omega-3 LCPUFA of terrestrial animal foods based on the application of measured response factors (RFs) to various FA. American mean intakes of pork, beef, chicken, and eggs contribute 20, 27, 45, and 71 mg/day of docosahexaenoic acid (DHA), respectively. The estimated intake of omega-3 DHA, eicosapentaenoic acid, and docosapentaenoic acid from nonseafood sources is significant, at 164, 103, and 330 mg/day, greater than most existing estimates of omega-3 LCPUFA intake.
Assuntos
Galinhas , Ovos , Ácidos Graxos Ômega-3 , Animais , Ácidos Graxos Ômega-3/análise , Ácidos Graxos Ômega-3/química , Ovos/análise , Humanos , Estados Unidos , Bovinos , Suínos , Carne/análise , Espectrometria de Massas em Tandem/métodos , Espectrometria de Massas/métodos , Ácidos Docosa-Hexaenoicos/análise , Ácidos Docosa-Hexaenoicos/químicaRESUMO
The dried root of Codonopsis pilosula (Franch.) Nannf., referred to as Dangshen in Chinese, is a famous traditional Chinese medicine. Polysaccharides, lobetyolin, and atractylenolide III are the major bioactive components contributing to its medicinal properties. Here, we investigated the dynamic changes of the main substances in annual Dangshen harvested at 12 time points from 20 May to 20 November 2020 (from early summer to early winter). Although the root biomass increased continuously, the crude polysaccharides content increased and then declined as the temperature fell, and so did the content of soluble proteins. However, the content of total phenolics and flavonoids showed an opposite trend, indicating that the carbon flux was changed between primary metabolism and secondary metabolism as the temperature and growth stages changed. The changes in the contents of lobetyolin and atractylenolide III indicated that autumn might be a suitable harvest time for Dangshen. The antioxidant capacity in Dangshen might be correlated with vitamin C. Furthermore, we analyzed the expression profiles of a few enzyme genes involved in the polysaccharide biosynthesis pathways at different growth stages, showing that CpUGpase and CPPs exhibited a highly positive correlation. These results might lay a foundation for choosing cultivars using gene expression levels as markers.
RESUMO
Sea urchin (class Echinoidea) gonads are a prized delicacy in Japan and many other world cultures. The complexity of its fatty acid (FA) profile, particularly minor FA, presents a formidable analytical challenge. We applied solvent mediated (SM) covalent adduct chemical ionization (CACI) tandem mass spectrometry to comprehensive de novo structural and quantitative characterization of the FA profile of Gulf of Mexico Atlantic sea urchin (Arbacia punctulata). >100 FA were detected including many with unusual double bond structure. Gulf sea urchin gonad lipids are rich in Δ5 monounsaturated FA 20:1(5Z) at 2.7% and the polymethylene-interrupted (PMI) diene 20:2(5Z,11Z) at 4.9%, as well as common omega-3 eicosapentaenoic acid (EPA; 5Z, 8Z, 11Z, 14Z, 17Z) at 9.8%±3.1% and arachidonic acid (AA; 5Z, 8Z, 11Z, 14Z) at 6.1%±2.1%. We propose plausible desaturation/elongation-based biochemical pathways for the endogenous production of unusual unsaturates. Unusual unsaturates may modify mammalian signaling and present novel bioactivities.
Assuntos
Arbacia , Ácidos Graxos Ômega-3 , Animais , Ácidos Graxos , Golfo do México , Espectrometria de Massas , Ouriços-do-Mar , SolventesRESUMO
Laccase (LAC) is a blue multicopper oxidase that contains four copper ions, which is involved in lignin polymerization and flavonoid biosynthesis in plants. Although dozens of LAC genes have been identified in Salvia miltiorrhiza Bunge (a model medicinal plant), most have not been functionally characterized. Here, we explored the expression patterns and the functionality of SmLAC25 in S. miltiorrhiza. SmLAC25 has a higher expression level in roots and responds to methyl jasmonate, auxin, abscisic acid, and gibberellin stimuli. The SmLAC25 protein is localized in the cytoplasm and chloroplasts. Recombinant SmLAC25 protein could oxidize coniferyl alcohol and sinapyl alcohol, two monomers of G-lignin and S-lignin. To investigate its function, we generated SmLAC25-overexpressed S. miltiorrhiza plantlets and hairy roots. The lignin content increased significantly in all SmLAC25-overexpressed plantlets and hairy roots, compared with the controls. However, the concentrations of rosmarinic acid and salvianolic acid B decreased significantly in all the SmLAC25-overexpressed lines. Further studies revealed that the transcription levels of some key enzyme genes in the lignin synthesis pathway (e.g., SmCCR and SmCOMT) were significantly improved in the SmLAC25-overexpressed lines, while the expression levels of multiple enzyme genes in the salvianolic acid biosynthesis pathway were inhibited. We speculated that the overexpression of SmLAC25 promoted the metabolic flux of lignin synthesis, which resulted in a decreased metabolic flux to the salvianolic acid biosynthesis pathway.
Assuntos
Salvia miltiorrhiza , Salvia miltiorrhiza/genética , Salvia miltiorrhiza/metabolismo , Lignina/metabolismo , Alcenos/metabolismo , Polifenóis/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
The papillary thyroid carcinoma (PTC) metastasizes through lymphatic spread, but the follicular thyroid cancer (FTC) metastasis occurs by following hematogenous spread. To date, the molecular mechanism underlying different metastatic routes between PTC and FTC is still unclear. Here, we showed that specifically androgen-regulated gene (SARG) was significantly up-regulated in PTC, while obviously down-regulated in FTC through analyzing the Gene Expression Omnibus (GEO) database. Immunohistochemistry assay verified that the PTC lymph node metastasis was associated with higher levels of SARG protein in clinical PTC patient samples. SARG-knockdown decreased TPC-1 and CGTH-W3 cells viability and migration significantly. On the contrary, SARG-overexpressed PTC cells possessed more aggressive migratory ability and viability. In vivo, SARG overexpression dramatically promoted popliteal lymph node metastasis of xenografts from TPC-1 cells mouse footpad transplanting. Mechanistically, SARG overexpression and knockdown significantly increased and decreased the expression of vascular endothelial growth factor C (VEGF-C) and VEGF receptor 3 (VEGFR-3), respectively, thereby facilitating or inhibiting the tube formation in HUVECs. The tube formation experiment showed that SARG overexpression and knockdown promoted or inhibited the number of tube formations in HUVEC cells, respectively. Taken together, we showed for the first time the differential expression profile of SARG between PTC and FTC, and SARG promotes PTC lymphatic metastasis via VEGF-C/VEGFR-3 signal. It indicates that SARG may represent a target for clinical intervention in lymphatic metastasis of PTC.
RESUMO
OBJECTIVE: To evaluate the hemodynamic state of critically ill cancer patients using central venous-to-arterial carbon dioxide difference (Pcv-aCO(2)) and to direct the treatment. METHODS: Clinical data of 47 cancer critically ill patients with acute physiology and chronic health evaluation II (APACHE II) score> 15 and unstable hemodynamic state were enrolled from intensive care unit of Tianjin Medical University Cancer Hospital from October 1st 2010 to May 31th 2011, were analyzed retrospectively. The patients were receiving the standard treatment according to the guidelines for 24 hours, the end-point of therapy was the standard of early goal direct therapy (EGDT). According to difference of sequential organ failure scores (ΔSOFA) of that after treatment and before treatment, the patients were divided into two groups: ΔSOFA≤ 1 (n = 27) and ΔSOFA> 1 (n = 20). The mean arterial pressure (MAP), urine output per hour, central venous pressure (CVP), oxygen saturation of central venous blood (ScvO(2)), the clearance of lactic acid, and Pcv-aCO(2) before treatment were compared with those after treatment, and their correlation with ΔSOFA was analysed. RESULTS: There were no significant differences in MAP (mm Hg, 1 mm Hg = 0.133 kPa: 54.48 ± 4.95 vs. 54.45 ± 4.30), urine output per hour (ml:19.33 ± 4.53 vs. 20.55 ± 5.54), CVP(mm Hg: 3.48 ± 1.81 vs. 3.25 ± 1.16), ScvO(2) (0.571 ± 0.042 vs. 0.578 ± 0.047) of two groups before treatment (all P > 0.05), but in the group ΔSOFA≤1, the Pcv-aCO(2) (mm Hg: 7.80 ± 2.20 vs. 9.39 ± 0.97) and SOFA scores (6.33 ± 2.11 vs. 9.50 ± 1.24) were significantly lower than those of the group ΔSOFA>1 (all P < 0.01). There were no significant differences in MAP (mm Hg: 73.48 ± 6.12 vs. 71.30 ± 7.30), CVP (mm Hg: 6.85 ± 1.26 vs. 6.50 ± 1.28), ScvO(2) (0.693 ± 0.032 vs. 0.684 ± 0.039) between two groups after treatment (all P > 0.05), though their RESULTS: data were improved compared with that of before treatment. However, there were significant differences in Pcv-aCO(2) (mm Hg: 3.02 ± 1.59 vs. 8.21 ± 2.23), urine output per hour (ml: 71.41 ± 6.74 vs. 51.70 ± 7.50), SOFA score (6.03 ± 2.56 vs. 10.05 ± 1.61), the clearance of lactic acid [(27.71 ± 11.46)% vs. -(0.78 ± 13.29)%, all P < 0.01]. There was significant correlation between urine output per hour, Pcv-aCO(2), clearance of lactic acid and ΔSOFA (r values were -0.712, 0.745, -0.631, all P < 0.05). CONCLUSION: Pcv-aCO(2) could be used as an index of evaluating the cardiac index and the hemodynamic state, and it could be considered to be one of the indices of evaluating the therapeutic effect and prognosis.