Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 46
Filtrar
1.
Cell Mol Life Sci ; 80(6): 167, 2023 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-37249637

RESUMO

Monosialoganglioside GM3 is the simplest ganglioside involved in various cellular signaling. Cell surface distribution of GM3 is thought to be crucial for the function of GM3, but little is known about the cell surface GM3 distribution. It was shown that anti-GM3 monoclonal antibody binds to GM3 in sparse but not in confluent melanoma cells. Our model membrane study evidenced that monoclonal anti-GM3 antibodies showed stronger binding when GM3 was in less fluid membrane environment. Studies using fluorescent GM3 analogs suggested that GM3 was clustered in less fluid membrane. Moreover, fluorescent lifetime measurement showed that cell surface of high density melanoma cells is more fluid than that of low density cells. Lipidomics and fatty acid supplementation experiment suggested that monounsaturated fatty acid-containing phosphatidylcholine contributed to the cell density-dependent membrane fluidity. Our results indicate that anti-GM3 antibody senses GM3 clustering and the number and/or size of GM3 cluster differ between sparse and confluent melanoma cells.


Assuntos
Gangliosídeo G(M3) , Melanoma , Humanos , Gangliosídeo G(M3)/metabolismo , Membrana Celular/metabolismo , Anticorpos Monoclonais , Melanoma/metabolismo , Contagem de Células
2.
Cardiovasc Diabetol ; 19(1): 19, 2020 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-32070346

RESUMO

BACKGROUND: Empagliflozin (empa), a selective sodium-glucose cotransporter (SGLT)2 inhibitor, reduced cardiovascular mortality and hospitalization for heart failure in patients with type 2 diabetes at high cardiovascular risk independent of glycemic control. The cardiovascular protective effect of empa was evaluated in an experimental model of metabolic syndrome, the obese ZSF1 rat, and its' lean control. METHODS: Lean and obese ZSF1 rats were either non-treated or treated with empa (30 mg/kg/day) for 6 weeks. Vascular reactivity was assessed using mesenteric artery rings, systolic blood pressure by tail-cuff sphygmomanometry, heart function and structural changes by echocardiography, and protein expression levels by Western blot analysis. RESULTS: Empa treatment reduced blood glucose levels from 275 to 196 mg/dl in obese ZSF1 rats whereas normoglycemia (134 mg/dl) was present in control lean ZSF1 rats and was unaffected by empa. Obese ZSF1 rats showed increased systolic blood pressure, and blunted endothelium-dependent relaxations associated with the appearance of endothelium-dependent contractile responses (EDCFs) compared to control lean rats. These effects were prevented by the empa treatment. Obese ZSF1 rats showed increased weight of the heart and of the left ventricle volume without the presence of diastolic or systolic dysfunction, which were improved by the empa treatment. An increased expression level of senescence markers (p53, p21, p16), tissue factor, VCAM-1, SGLT1 and SGLT2 and a down-regulation of eNOS were observed in the aortic inner curvature compared to the outer one in the control lean rats, which were prevented by the empa treatment. In the obese ZSF1 rats, no such effects were observed. The empa treatment reduced the increased body weight and weight of lungs, spleen, liver and perirenal fat, hyperglycemia and the increased levels of total cholesterol and triglycerides in obese ZSF1 rats, and increased blood ketone levels and urinary glucose excretion in control lean and obese ZSF1 rats. CONCLUSION: Empa reduced glucose levels by 28% and improved both endothelial function and cardiac remodeling in the obese ZSF1 rat. Empa also reduced the increased expression level of senescence, and atherothrombotic markers at arterial sites at risk in the control lean, but not obese, ZSF1 rat.


Assuntos
Compostos Benzidrílicos/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Glucosídeos/farmacologia , Síndrome Metabólica/tratamento farmacológico , Inibidores do Transportador 2 de Sódio-Glicose/farmacologia , Função Ventricular Esquerda/efeitos dos fármacos , Remodelação Ventricular/efeitos dos fármacos , Animais , Biomarcadores/sangue , Glicemia/efeitos dos fármacos , Glicemia/metabolismo , Senescência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Endotélio Vascular/metabolismo , Endotélio Vascular/fisiopatologia , Síndrome Metabólica/etiologia , Síndrome Metabólica/metabolismo , Síndrome Metabólica/fisiopatologia , Obesidade/complicações , Ratos Zucker , Sístole
3.
Int J Mol Sci ; 21(3)2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32019237

RESUMO

In humans, aging is associated with endothelial dysfunction and an increased risk of venous thromboembolism. Although intake of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) at a ratio of 6:1 by old rats improved the endothelial dysfunction in arteries, the impact on veins remains unclear. Eight-month-old male Wistar rats were either untreated or orally administered corn oil, EPA:DHA 1:1, or EPA:DHA 6:1 (500 mg/kg/d) for seven days. Vascular reactivity was studied by myography. In middle-aged femoral artery rings, acetylcholine caused a partial relaxation at low concentrations and a contractile response at high concentrations, whereas in the old femoral vein only a partial relaxation was observed. The EPA:DHA 6:1 treatment blunted the contractile response to acetylcholine in the middle-aged femoral artery and both EPA:DHA 6:1 and 1:1 increased the relaxation to acetylcholine in the old femoral vein. No such effects were observed with corn oil. Both the non-selective cyclooxygenase inhibitor indomethacin and the COX-1 inhibitor SC-560 increased the relaxation to acetylcholine in the middle-aged femoral artery whereas the COX-2 inhibitor NS-398 increased that in the middle-aged femoral vein. In conclusion, our results indicate that aging is associated with an endothelial dysfunction in the femoral artery and vein, which can be improved by EPA:DHA 6:1 treatment-most likely via a cyclooxygenase-dependent mechanism.


Assuntos
Envelhecimento/patologia , Ácidos Docosa-Hexaenoicos/administração & dosagem , Ácido Eicosapentaenoico/administração & dosagem , Endotélio Vascular/efeitos dos fármacos , Artéria Femoral/efeitos dos fármacos , Veia Femoral/efeitos dos fármacos , Prostaglandina-Endoperóxido Sintases/química , Doenças Vasculares/tratamento farmacológico , Administração Oral , Animais , Inibidores de Ciclo-Oxigenase/farmacologia , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Artéria Femoral/metabolismo , Artéria Femoral/patologia , Veia Femoral/metabolismo , Veia Femoral/patologia , Masculino , Ratos , Ratos Wistar , Doenças Vasculares/metabolismo , Doenças Vasculares/patologia
4.
Plant Cell ; 23(3): 1124-37, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21447792

RESUMO

Peroxidases have been shown to be involved in the polymerization of lignin precursors, but it remains unclear whether laccases (EC 1.10.3.2) participate in constitutive lignification. We addressed this issue by studying laccase T-DNA insertion mutants in Arabidopsis thaliana. We identified two genes, LAC4 and LAC17, which are strongly expressed in stems. LAC17 was mainly expressed in the interfascicular fibers, whereas LAC4 was expressed in vascular bundles and interfascicular fibers. We produced two double mutants by crossing the LAC17 (lac17) mutant with two LAC4 mutants (lac4-1 and lac4-2). The single and double mutants grew normally in greenhouse conditions. The single mutants had moderately low lignin levels, whereas the stems of lac4-1 lac17 and lac4-2 lac17 mutants had lignin contents that were 20 and 40% lower than those of the control, respectively. These lower lignin levels resulted in higher saccharification yields. Thioacidolysis revealed that disrupting LAC17 principally affected the deposition of G lignin units in the interfascicular fibers and that complementation of lac17 with LAC17 restored a normal lignin profile. This study provides evidence that both LAC4 and LAC17 contribute to the constitutive lignification of Arabidopsis stems and that LAC17 is involved in the deposition of G lignin units in fibers.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Lacase/genética , Lignina/biossíntese , Caules de Planta/metabolismo , Feixe Vascular de Plantas/genética , Arabidopsis/enzimologia , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Teste de Complementação Genética , Inflorescência/genética , Inflorescência/metabolismo , Lacase/isolamento & purificação , Lacase/metabolismo , Lignina/análise , Lignina/genética , Mutação , Caules de Planta/anatomia & histologia , Caules de Planta/crescimento & desenvolvimento , Feixe Vascular de Plantas/enzimologia , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas
5.
Methods Enzymol ; 700: 217-234, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38971601

RESUMO

Sphingomyelin is postulated to form clusters with glycosphingolipids, cholesterol and other sphingomyelin molecules in biomembranes through hydrophobic interaction and hydrogen bonds. These clusters form submicron size lipid domains. Proteins that selectively binds sphingomyelin and/or cholesterol are useful to visualize the lipid domains. Due to their small size, visualization of lipid domains requires advanced microscopy techniques in addition to lipid binding proteins. This Chapter describes the method to characterize plasma membrane sphingomyelin-rich and cholesterol-rich lipid domains by quantitative microscopy. This Chapter also compares different permeabilization methods to visualize intracellular lipid domains.


Assuntos
Colesterol , Esfingomielinas , Esfingomielinas/química , Esfingomielinas/metabolismo , Colesterol/química , Colesterol/metabolismo , Humanos , Animais , Microdomínios da Membrana/metabolismo , Microdomínios da Membrana/química , Microscopia/métodos , Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Membrana Celular/metabolismo , Membrana Celular/química
6.
Nat Commun ; 14(1): 7353, 2023 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-37990014

RESUMO

Although the human immunodeficiency virus type 1 lipid envelope has been reported to be enriched with host cell sphingomyelin and cholesterol, the molecular mechanism of the enrichment is not well understood. Viral Gag protein plays a central role in virus budding. Here, we report the interaction between Gag and host cell lipids using different quantitative and super-resolution microscopy techniques in combination with specific probes that bind endogenous sphingomyelin and cholesterol. Our results indicate that Gag in the inner leaflet of the plasma membrane colocalizes with the outer leaflet sphingomyelin-rich domains and cholesterol-rich domains, enlarges sphingomyelin-rich domains, and strongly restricts the mobility of sphingomyelin-rich domains. Moreover, Gag multimerization induces sphingomyelin-rich and cholesterol-rich lipid domains to be in close proximity in a curvature-dependent manner. Our study suggests that Gag binds, coalesces, and reorganizes pre-existing lipid domains during assembly.


Assuntos
HIV-1 , Humanos , HIV-1/metabolismo , Esfingomielinas/metabolismo , Membrana Celular/metabolismo , Produtos do Gene gag/metabolismo , Colesterol/metabolismo , Microdomínios da Membrana/metabolismo
7.
Plant Biotechnol J ; 10(5): 609-20, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22458713

RESUMO

Lignocellulosic biomass is utilized as a renewable feedstock in various agro-industrial activities. Lignin is an aromatic, hydrophobic and mildly branched polymer integrally associated with polysaccharides within the biomass, which negatively affects their extraction and hydrolysis during industrial processing. Engineering the monomer composition of lignins offers an attractive option towards new lignins with reduced recalcitrance. The presented work describes a new strategy developed in Arabidopsis for the overproduction of rare lignin monomers to reduce lignin polymerization degree (DP). Biosynthesis of these 'DP reducers' is achieved by expressing a bacterial hydroxycinnamoyl-CoA hydratase-lyase (HCHL) in lignifying tissues of Arabidopsis inflorescence stems. HCHL cleaves the propanoid side-chain of hydroxycinnamoyl-CoA lignin precursors to produce the corresponding hydroxybenzaldehydes so that plant stems expressing HCHL accumulate in their cell wall higher amounts of hydroxybenzaldehyde and hydroxybenzoate derivatives. Engineered plants with intermediate HCHL activity levels show no reduction in total lignin, sugar content or biomass yield compared with wild-type plants. However, cell wall characterization of extract-free stems by thioacidolysis and by 2D-NMR revealed an increased amount of unusual C6C1 lignin monomers most likely linked with lignin as end-groups. Moreover the analysis of lignin isolated from these plants using size-exclusion chromatography revealed a reduced molecular weight. Furthermore, these engineered lines show saccharification improvement of pretreated stem cell walls. Therefore, we conclude that enhancing the biosynthesis and incorporation of C6C1 monomers ('DP reducers') into lignin polymers represents a promising strategy to reduce lignin DP and to decrease cell wall recalcitrance to enzymatic hydrolysis.


Assuntos
Arabidopsis/metabolismo , Hidroliases/metabolismo , Lignina/biossíntese , Caules de Planta/metabolismo , Arabidopsis/genética , Biomassa , Parede Celular/metabolismo , Regulação da Expressão Gênica de Plantas , Hidroliases/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Polimerização , Regiões Promotoras Genéticas , Transformação Genética
8.
J Exp Bot ; 63(11): 4291-301, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22553285

RESUMO

Trees will have to cope with increasing levels of CO(2) and ozone in the atmosphere. The purpose of this work was to assess whether the lignification process could be altered in the wood of poplars under elevated CO(2) and/or ozone. Young poplars were exposed either to charcoal-filtered air (control), to elevated CO(2) (800 µl l(-1)), to ozone (200 nl l(-1)) or to a combination of elevated CO(2) and ozone in controlled chambers. Lignification was analysed at different levels: biosynthesis pathway activities (enzyme and transcript), lignin content, and capacity to incorporate new assimilates by using (13)C labelling. Elevated CO(2) and ozone had opposite effects on many parameters (growth, biomass, cambial activity, wood cell wall thickness) except on lignin content which was increased by elevated CO(2) and/or ozone. However, this increased lignification was due to different response mechanisms. Under elevated CO(2), carbon supply to the stem and effective lignin synthesis were enhanced, leading to increased lignin content, although there was a reduction in the level of some enzyme and transcript involved in the lignin pathway. Ozone treatment induced a reduction in carbon supply and effective lignin synthesis as well as transcripts from all steps of the lignin pathway and some corresponding enzyme activities. However, lignin content was increased under ozone probably due to variations in other major components of the cell wall. Both mechanisms seemed to coexist under combined treatment and resulted in a high increase in lignin content.


Assuntos
Dióxido de Carbono/metabolismo , Lignina/metabolismo , Ozônio/metabolismo , Populus/metabolismo , Madeira/metabolismo , Populus/crescimento & desenvolvimento , Madeira/crescimento & desenvolvimento
9.
J Exp Bot ; 62(10): 3575-86, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21357770

RESUMO

Wood formation in trees is a dynamic process that is strongly affected by environmental factors. However, the impact of ozone on wood is poorly documented. The objective of this study was to assess the effects of ozone on wood formation by focusing on the two major wood components, cellulose and lignin, and analysing any anatomical modifications. Young hybrid poplars (Populus tremula × alba) were cultivated under different ozone concentrations (50, 100, 200, and 300 l l(-1)). As upright poplars usually develop tension wood in a non-set pattern, the trees were bent in order to induce tension wood formation on the upper side of the stem and normal or opposite wood on the lower side. Biosynthesis of cellulose and lignin (enzymes and RNA levels), together with cambial growth, decreased in response to ozone exposure. The cellulose to lignin ratio was reduced, suggesting that cellulose biosynthesis was more affected than that of lignin. Tension wood was generally more altered than opposite wood, especially at the anatomical level. Tension wood may be more susceptible to reduced carbon allocation to the stems under ozone exposure. These results suggested a coordinated regulation of cellulose and lignin deposition to sustain mechanical strength under ozone. The modifications of the cellulose to lignin ratio and wood anatomy could allow the tree to maintain radial growth while minimizing carbon cost.


Assuntos
Celulose/metabolismo , Lignina/metabolismo , Ozônio/farmacologia , Populus/metabolismo , Madeira/metabolismo , Populus/efeitos dos fármacos
10.
Plant J ; 58(2): 246-59, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19077165

RESUMO

BAHD acyltransferases catalyze the acylation of many plant secondary metabolites. We characterized the function of At2g19070, a member of the BAHD gene family of Arabidopsis thaliana. The acyltransferase gene was shown to be specifically expressed in anther tapetum cells in the early stages of flower development. The impact of gene repression was studied in RNAi plants and in a knockout (KO) mutant line. Immunoblotting with a specific antiserum raised against the recombinant protein was used to evaluate the accumulation of At2g19070 gene product in flowers of various Arabidopsis genotypes including the KO and RNAi lines, the male sterile mutant ms1 and transformants overexpressing the acyltransferase gene. Metabolic profiling of flower bud tissues from these genetic backgrounds demonstrated a positive correlation between the accumulation of acyltransferase protein and the quantities of metabolites that were putatively identified by tandem mass spectrometry as N(1),N(5),N(10)-trihydroxyferuloyl spermidine and N(1),N(5)-dihydroxyferuloyl-N(10)-sinapoyl spermidine. These products, deposited in pollen coat, can be readily extracted by pollen wash and were shown to be responsible for pollen autofluorescence. The activity of the recombinant enzyme produced in bacteria was assayed with various hydroxycinnamoyl-CoA esters and polyamines as donor and acceptor substrates, respectively. Feruloyl-CoA and spermidine proved the best substrates, and the enzyme has therefore been named spermidine hydroxycinnamoyl transferase (SHT). A methyltransferase gene (At1g67990) which co-regulated with SHT during flower development, was shown to be involved in the O-methylation of spermidine conjugates by analyzing the consequences of its repression in RNAi plants and by characterizing the methylation activity of the recombinant enzyme.


Assuntos
Aciltransferases/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Flores/enzimologia , Espermidina/biossíntese , Aciltransferases/genética , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Flores/genética , Regulação da Expressão Gênica de Plantas , Técnicas de Inativação de Genes , Metaboloma , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Pólen/metabolismo , Interferência de RNA , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
11.
New Phytol ; 184(1): 99-113, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19674336

RESUMO

A cinnamoyl-CoA reductase 1 knockout mutant in Arabidopsis thaliana was investigated for the consequences of lignin synthesis perturbation on the assembly of the cell walls. The mutant displayed a dwarf phenotype and a strong collapse of its xylem vessels corresponding to lower lignin content and a loss of lignin units of the noncondensed type. Transmission electron microscopy revealed that the transformation considerably impaired the capacity of interfascicular fibers and vascular bundles to complete the assembly of cellulose microfibrils in the S(2) layer, the S(1) layer remaining unaltered. Such disorder in cellulose was correlated with X-ray diffraction showing altered organization. Semi-quantitative immunolabeling of lignins showed that the patterns of distribution were differentially affected in interfascicular fibers and vascular bundles, pointing to the importance of noncondensed lignin structures for the assembly of a coherent secondary wall. The use of laser capture microdissection combined with the microanalysis of lignins and polysaccharides allowed these polymers to be characterized into specific cell types. Wild-type A. thaliana displayed a two-fold higher syringyl to guaiacyl ratio in interfascicular fibers compared with vascular bundles, whereas this difference was less marked in the cinnamoyl-CoA reductase 1 knockout mutant.


Assuntos
Aldeído Oxirredutases/genética , Arabidopsis/enzimologia , Parede Celular/enzimologia , Inativação Gênica , Lignina/metabolismo , Arabidopsis/ultraestrutura , Metabolismo dos Carboidratos , Parede Celular/ultraestrutura , Microanálise por Sonda Eletrônica , Flores/química , Flores/citologia , Flores/enzimologia , Flores/ultraestrutura , Cromatografia Gasosa-Espectrometria de Massas , Glucose/metabolismo , Imuno-Histoquímica , Lignina/química , Espectroscopia de Ressonância Magnética , Microdissecção , Mutação/genética , Extratos Vegetais/química , Caules de Planta/química , Caules de Planta/citologia , Caules de Planta/enzimologia , Caules de Planta/ultraestrutura , Coloração e Rotulagem , Difração de Raios X , Xilose/metabolismo
12.
Sci Rep ; 9(1): 5812, 2019 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-30967612

RESUMO

Ceramide phosphoethanolamine (CPE), a major sphingolipid in invertebrates, is crucial for axonal ensheathment in Drosophila. Darkfield microscopy revealed that an equimolar mixture of bovine buttermilk CPE (milk CPE) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (diC18:1 PC) tends to form tubules and helical ribbons, while pure milk CPE mainly exhibits amorphous aggregates and, at low frequency, straight needles. Negative staining electron microscopy indicated that helices and tubules were composed of multilayered 5-10 nm thick slab-like structures. Using different molecular species of PC and CPE, we demonstrated that the acyl chain length of CPE but not of PC is crucial for the formation of tubules and helices in equimolar mixtures. Incubation of the lipid suspensions at the respective phase transition temperature of CPE facilitated the formation of both tubules and helices, suggesting a dynamic lipid rearrangement during formation. Substituting diC18:1 PC with diC18:1 PE or diC18:1 PS failed to form tubules and helices. As hydrated galactosylceramide (GalCer), a major lipid in mammalian myelin, has been reported to spontaneously form tubules and helices, it is believed that the ensheathment of axons in mammals and Drosophila is based on similar physical processes with different lipids.


Assuntos
Drosophila/metabolismo , Galactosilceramidas/metabolismo , Membranas/química , Fosfatidilcolinas/metabolismo , Esfingomielinas/metabolismo , Animais , Fasciculação Axônica/fisiologia , Bicamadas Lipídicas/química , Conformação Molecular , Sistema Nervoso/metabolismo , Transição de Fase
13.
Proc Biol Sci ; 275(1648): 2221-9, 2008 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-18595839

RESUMO

Palms can grow at sites exposed to high winds experiencing large dynamic wind and gust loads. Their stems represent a system of stiff fibrous elements embedded in the soft parenchymatous tissue. The proper design of the interface of the stiffening elements and the parenchyma is crucial for the functioning of the stem. The strategy of the palm to compromise between stiff fibre caps and the soft parenchymatous tissue may serve as a model system for avoiding stress discontinuities in inhomogeneous and anisotropic fibre-reinforced composite materials. We investigated the mechanical, structural and biochemical properties of the fibre caps of the palm Washingtonia robusta at different levels of hierarchy with high spatial resolution. A gradual decrease in stiffness across the fibre cap towards the surrounding parenchymatous tissue was observed. Structural adaptations at the tissue level were found in terms of changes in cell cross sections and cell wall thickness. At the cell wall level, gradients across the fibre cap were found in the degree of orientation of the microfibrils and in the lignin level and composition. The impact of these structural variations in the local material stiffness distribution is discussed.


Assuntos
Arecaceae/fisiologia , Tecido Conjuntivo/fisiologia , Cromatografia Gasosa-Espectrometria de Massas , Espectrofotometria Ultravioleta , Resistência à Tração , Difração de Raios X
14.
Appl Microbiol Biotechnol ; 80(4): 719-33, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18654772

RESUMO

Proteomic analysis was performed to determine and differentiate the composition of the secretomes of Phanerochaete chrysosporium CIRM-BRFM41, a peroxidase hypersecretory strain grown under ligninolytic conditions and on softwood chips under biopulping conditions. Extracellular proteins from both cultures were analyzed by bidimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry. A total of 37 spots were identified. The secretome in liquid synthetic medium comprised mainly peroxidases, while several wood-degrading enzymes and enzymes involved in fungal metabolism were detected in biopulping cultures on softwood. This prompted an analysis of the impact of secretome modulation in the presence of softwood chips. Biotreated wood was submitted to kraft cooking and chemical bleaching using chlorine dioxide. The fungal pre-treatment led to a significant increase in pulp yield and a better bleachability of the pulp. This bleachability improvement could be explained by the production of specific lignocellulose-degrading enzymes.


Assuntos
Espaço Extracelular/metabolismo , Proteínas Fúngicas/metabolismo , Microbiologia Industrial , Lignina/metabolismo , Phanerochaete/metabolismo , Proteômica , Madeira/metabolismo , Sequência de Aminoácidos , Eletroforese em Gel Bidimensional , Espaço Extracelular/química , Espaço Extracelular/genética , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Lignina/química , Dados de Sequência Molecular , Phanerochaete/química , Phanerochaete/genética , Transporte Proteico , Madeira/química
15.
Tree Physiol ; 28(11): 1619-28, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18765367

RESUMO

From June to December, we determined the effects of variations in biochemical composition on delta(13)C of tree rings of 2-year-old oaks (Quercus petraea (Matt.) Liebl.) growing under semi-natural conditions, and the dependence of these effects of water stress during the growth season. Percent abundance, carbon concentration and delta(13)C were measured in holocellulose, lignin, extractive-free wood, starch and a water-soluble fraction. Relative concentrations of lignin and holocellulose in the extractive-free wood varied little during the season or in response to water stress, indicating that these compounds could not quantitatively explain the variations in whole-wood delta(13)C. Among all sampled tree rings, the relative concentration of each structural compound (holocellulose and lignin) accounted for less than 5% of the delta(13)C variability in whole wood. Variations in holocellulose and extractive-free wood delta(13)C between tree rings were almost identical (r > 0.95), whereas variations in lignin delta(13)C were less well correlated to these compounds. Whole-wood delta(13)C had a slightly altered isotopic signal compared with that of the structural compounds because of the presence of the extractive component. These results showed that variations in lignin delta(13)C and lignin concentration have little influence on extractive-free wood delta(13)C and whole-wood delta(13)C. Rather, holocellulose influences delta(13)C the most. Thus, we confirmed that, for climatic reconstruction from tree rings, removal of extractives by soxhlet is generally sufficient and sometimes unnecessary. Our findings also indicate that, in the case of rapid and severe water stress, the structural component did not accurately record the associated increase in delta(13)C because of dilution with previously formed organic matter and cessation of trunk growth. The effect of drought on carbon isotope ratios was more pronounced in the extractive compounds, making them good water stress indicators but only on a scale of days to months.


Assuntos
Carbono/química , Quercus/crescimento & desenvolvimento , Quercus/metabolismo , Carbono/metabolismo , Isótopos de Carbono , Lignina/metabolismo , Estações do Ano , Água/metabolismo , Madeira/química , Madeira/metabolismo
16.
Nutrition ; 51-52: 53-59, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29605764

RESUMO

OBJECTIVE: Polyphenol-rich products such as fruit juices have been found to have strong antioxidant capacities and to induce potent endothelium-dependent relaxation. We evaluated whether the commercial blackcurrant juices induced endothelium-dependent relaxation of isolated coronary arteries can be related to their antioxidant capacity and/or phenolic content. METHODS: Six different commercial blackcurrant juices were selected. Their main phenolic compounds were measured by ultra-performance liquid chromatography and antioxidant capacity was evaluated by spectrometric methods. Vascular reactivity studies with these juices were done using isolated porcine coronary arteries. RESULTS: The six different commercial blackcurrant juices induced relaxation ranging from 21% to 100% at the concentration of 0.5% volume per volume (v/v). The relaxation induced at 0.5% v/v was not correlated to their antioxidant capacity measured by either oxygen radical antioxidant capacity or DPPH (2,2-diphenyl-1-picrylhydrazyl) assays and also not to the ascorbic acid, total polyphenols, total flavanols, and total phenolic acid contents. In contrast, the amplitude of the relaxation was correlated to the total anthocyanins content and the individual anthocyanin concentration. CONCLUSIONS: Correlations between relaxation amplitude and total anthocyanin or individual anthocyanin contents are of interest for the development of functional blackcurrant beverages with the potential to promote vascular protection.


Assuntos
Antocianinas/farmacologia , Antioxidantes , Vasos Coronários/efeitos dos fármacos , Sucos de Frutas e Vegetais , Grossulariaceae , Vasodilatação/efeitos dos fármacos , Animais , Vasos Coronários/fisiologia , Frutas , Suínos
17.
Phytochemistry ; 68(4): 571-9, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17187834

RESUMO

The objective of this study was to assess the influence of the peroxidase/coniferyl alcohol (CA) ratio on the dehydrogenation polymer (DHP) synthesis. The soluble and unsoluble fractions of horseradish peroxidase (HRP)-catalyzed CA dehydrogenation mixtures were recovered in various proportions, depending on the polymerization mode (Zutropf ZT/Zulauf ZL) and HRP/CA ratio (1.6-1100purpurogallin U mmol(-1)). The ZL mode yielded 0-57%/initial CA of unsoluble condensed DHPs (thioacidolysis yields <200micromolg(-1)) with a proportion of uncondensed CA end groups increasing with the HRP/CA ratio (7.2-55.5%/total uncondensed CA). Systematically lower polymer yields (0-49%/initial CA) were obtained for the ZT mode. In that mode, a negative correlation was established between the beta-O-4 content (thioacidolysis yields: 222-660micromolg(-1)) and the HRP/CA ratio. In both modes, decreasing the HRP/CA ratio below 18Ummol(-1) favoured an end-wise polymerization process evidenced by the occurrence of tri-, tetra- and pentamers involving at least one beta-O-4 bond. At low ratio, the unsoluble ZT DHP was found to better approximate natural lignins than DHPs previously synthesized with traditional methods. Besides its possible implication in lignin biosynthesis, peroxidase activity is a crucial parameter accounting for the structural variations of in vitro DHPs.


Assuntos
Peroxidase do Rábano Silvestre/metabolismo , Lignina/química , Lignina/metabolismo , Benzocicloeptenos/metabolismo , Cromatografia Líquida , Cinética , Espectrometria de Massas , Oxirredução
18.
Nat Biotechnol ; 20(6): 607-12, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12042866

RESUMO

The agronomic and pulping performance of transgenic trees with altered lignin has been evaluated in duplicated, long-term field trials. Poplars expressing cinnamyl alcohol dehydrogenase (CAD) or caffeate/5-hydroxy-ferulate O-methyltransferase (COMT) antisense transgenes were grown for four years at two sites, in France and England. The trees remained healthy throughout the trial. Growth indicators and interactions with insects were normal. No changes in soil microbial communities were detected beneath the transgenic trees. The expected modifications to lignin were maintained in the transgenics over four years, at both sites. Kraft pulping of tree trunks showed that the reduced-CAD lines had improved characteristics, allowing easier delignification, using smaller amounts of chemicals, while yielding more high-quality pulp. This work highlights the potential of engineering wood quality for more environmentally benign papermaking without interfering with tree growth or fitness.


Assuntos
Lignina/genética , Lignina/metabolismo , Plantas Geneticamente Modificadas , Populus/genética , Populus/metabolismo , Madeira , Oxirredutases do Álcool/genética , Oxirredutases do Álcool/metabolismo , Elementos Antissenso (Genética) , Ecossistema , Poluição Ambiental/prevenção & controle , França , Expressão Gênica , Resíduos Industriais/prevenção & controle , Metiltransferases/genética , Metiltransferases/metabolismo , Modelos Químicos , Papel , Raízes de Plantas/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Microbiologia do Solo , Transgenes , Reino Unido
19.
J Agric Food Chem ; 55(25): 10306-14, 2007 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-17988087

RESUMO

The short-term influence of adequate and high nitrogen fertilization on poplar lignification was investigated. The high nitrogen supply decreased lignin staining in the newly formed secondary xylem, indicating that lignin deposition was affected. Acetyl bromide determinations gave a 9-10% decrease in lignin content; however, Klason lignin content was unchanged. Thioacidolysis showed that elevated N supply affected lignin structure such that there was a reduced frequency of lignin units involved in beta-O-4 bonds, a reduced syringyl/guaiacyl ratio, an increased frequency of p-hydroxyphenyl lignin units, more guaiacyl units with free phenolic groups, and more p-hydroxybenzoic acid ester-linked to poplar lignins. These features suggest that lignins from poplars grown under high N bear structural similarities to lignins formed during early stages of wood development. The findings also indicate that a gravitational stimulus inducing the formation of tension wood and high N availability lead to similar and additive effects on lignin content and structure.


Assuntos
Lignina/análise , Lignina/química , Nitrogênio/administração & dosagem , Populus/química , Populus/crescimento & desenvolvimento , Madeira/química , Fertilizantes
20.
J Agric Food Chem ; 53(15): 5872-81, 2005 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-16028968

RESUMO

Grass cell wall degradability is conventionally related to the lignin content and to the ferulic-mediated cross-linking of lignins to polysaccharides. To better understand the variations in degradability, 22 maize inbred lines were subjected to image analyses of Fasga- and Mäule-stained stem sections and to chemical analyses of lignins and p-hydroxycinnamic acids. For the first time, the nearness of biochemical and histological estimates of lignin levels was established. Combination of histological and biochemical traits could explain 89% of the variations for cell wall degradability and define a maize ideotype for cell wall degradability. In addition to a reduced lignin level, such an ideotype would contain lignins richer in syringyl than in guaiacyl units and preferentially localized in the cortical region rather than in the pith. Such enrichment in syringyl units would favor wall degradability in grasses, contrary to dicots, and could be related to the fact that grass syringyl units are noticeably p-coumaroylated. This might affect the interaction capabilities of lignins and polysaccharides.


Assuntos
Parede Celular/química , Parede Celular/metabolismo , Lignina/análise , Zea mays/ultraestrutura , Cruzamento , Variação Genética , Zea mays/química , Zea mays/genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA