Lycopene intervention reduces inflammation and improves HDL functionality in moderately overweight middle-aged individuals

The management of overweight subjects by interventions aimed at reducing inflammation is highly desirable. To date, observational studies have identified a link between increased dietary antioxidant intake and reduced cardiovascular morbidity. However, direct trial evidence regarding the ability of antioxidants to influence inflammation is lacking. Therefore, this study examined lycopene's ability to lower systemic and high-density lipoprotein (HDL)-associated inflammation in moderately overweight middle-aged subjects. Serum was collected before and after a 12-week intervention from 54 moderately overweight, middle-aged individuals. Subjects were randomised to one of three groups: control diet (

The effects of Chronic Lycopene Supplementation on Exercise-Induced Oxidative Stress and Glucose Homeostasis.

Lycopene can exert antioxidant effects against peripheral and cellular oxidative stress and may be associated with reduced diabetic risk. Conversely, exercise-induced free radicals are thought to underpin many of the desirable whole-body adaptations following training and the use of antioxidants within the exercise model remains debatable. PURPOSE: To investigate the effect of lycopene supplementation on oxidative stress and glucose homeostasis following acute aerobic exercise. METHOD: Twenty-eight (n=28) apparently healthy male volunteers were recruited (age 24 ± 4 years; weight 78 ± 10 kg; height 178 ± 8 cm; 2max 40 ± 7 ml·kg-1 ·min-1 ) in a randomised, single blind, placebo-controlled study. Participants were required to attend the Laboratory on two occasions: prior to and following 6 weeks of supplementation of either 10mg lycopene (LG; n=15) or placebo (PG; n=13) followed by a bout of acute exercise for one hour at 65% 2max. Exogenous glucose oxidation was then measured on an isotope ratio mass spectrometer in a sub-group of participants (n=14) following exercise, by administration of a standard oral glucose tolerance test (OGTT; 75g glucose). Venous blood samples were drawn for measurement of oxidative stress parameters, plasma glucose and insulin. RESULTS: Plasma lycopene increased in LG only (0.01 ± 0.004 vs.0.02 ± 0.007 µmol/L; P <0.05) following supplementation and remained elevated post exercise compared to PG (0.01 ± 0.004 vs. 0.02 ± 0.009 µmol/L; P <0.05). There were no changes in other markers of oxidative stress (SOD, LOOHs, F2 ISP and Alkoxyl radical) either between or within the trials, (P >0.05, respectively). A main effect for an increase in insulin was observed two hours post OGTT in the sub-groups (Pooled data, P <0.05) but trends in the HOMA scores were evident with a 57% increase for LG (2.20 ± 1.84 vs. 5.14 ± 2.5; P >0.05) and an 11% decrease for PG (2.17 ± 1.06 vs. 1.94 ± 1.53; P >0.05). No change in plasma glucose was detected at any point, or after the OGTT (P >0.05). CONCLUSION: In healthy males, lycopene supplementation had no effect on post exercise levels of ROS or markers of lipid peroxidation, despite an increase in plasma lycopene. However, lycopene supplementation may affect post exercise insulin sensitivity in response to glucose consumption, but further parallel research is required.

Chp8, a diguanylate cyclase from Pseudomonas syringae pv tomato DC3000 suppresses the PAMP flagellin, increases EPS and promotes plant immune evasion.

The bacterial plant pathogen Pseudomonas syringae causes disease in a wide range of plants. The associated decrease in crop yields results in economic losses and threatens global food security. Competition exists between the plant immune system and the pathogen, the basic principles of which can be applied to animal infection pathways. P. syringae uses a type III secretion system (T3SS) to deliver virulence factors into the plant that promote survival of the bacterium. The P. syringae T3SS is a product of the hypersensitive response and pathogenicity (hrp) and hypersensitive response and conserved (hrc) gene cluster, which is strictly controlled by the codependent enhancer-binding proteins HrpR and HrpS. Through a combination of bacterial gene regulation and phenotypic studies, plant infection assays, and plant hormone quantifications, we now report that Chp8 (i) is embedded in the Hrp regulon and expressed in response to plant signals and HrpRS, (ii) is a functional diguanylate cyclase, (iii) decreases the expression of the major pathogen-associated molecular pattern (PAMP) flagellin and increases extracellular polysaccharides (EPS), and (iv) impacts the salicylic acid/jasmonic acid hormonal immune response and disease progression. We propose that Chp8 expression dampens PAMP-triggered immunity during early plant infection.

Development of a sustainable method for the extraction of lycopene and β-carotene from food wastes

The main objective of the present work is the study of a profitable process not only in the extraction and selective separation of lycopene and β-carotene, two compounds present in tomato, but also in its potential application to food industry wastes. This is one of the industries that produce larger amounts of wastes, which are rich in high value biomolecules with great economic interest. However, the conventional methods used to extract this kind of compounds are expensive which limits their application at large scale. Lycopene and βcarotene are carotenoids with high commercial value, known for their antioxidant activity and benefits to human health. Their biggest source is tomato, one of the world’s most consumed fruits, reason for which large quantities of waste is produced. This work focuses on the study of diverse solvents with a high potential to extract carotenoids from tomato, as well as the search for more environmentally benign solvents than those currently used to extract lycopene and β-carotene from biomass. Additionally, special attention was paid to the creation of a continuous process that would allow the fractionation of the compounds for further purification. Thus, the present work started with the extraction of both carotenoids using a wide range of solvents, namely, organic solvents, conventional salts, ionic liquids, polymers and surfactants. In this stage, each solvent was evaluated in what regards their capacity of extraction as well as their penetration ability in biomass. The results collected showed that an adequate selection of the solvents may lead to the complete extraction of both carotenoids in one single step, particularly acetone and tetrahydrofuran were the most effective ones. However, the general low penetration capacity of salts, ionic liquids, polymers and surfactants makes these solvents ineffective in the solid-liquid extraction process. As the organic solvents showed the highest capacity to extract lycopene and βcarotene, in particular tetrahydrofuran and acetone, the latter solvent used in the development process of fractionation, using to this by strategic use of solvents. This step was only successfully developed through the manipulation of the solubility of each compound in ethanol and n-hexane. The results confirmed the possibility of fractionating the target compounds using the correct addition order of the solvents. Approximately, 39 % of the β-carotene was dissolved in ethanol and about 64 % of lycopene was dissolved in n-hexane, thus indicating their separation for two different solvents which shows the selective character of the developed process without any prior stage optimization. This study revealed that the use of organic solvents leads to selective extraction of lycopene and β-carotene, allowing diminishing the numerous stages involved in conventional methods. At the end, it was possible to idealize a sustainable and of high industrial relevance integrated process, nevertheless existing the need for additional optimization studies in the future.

GREBP, un nouveau facteur de transcription contrôlant l’expression de la guanylate cyclase A, récepteur de l’ANP, via l’élément de réponse au cGMP

La découverte du système des peptides natriurétiques (NP), au début des années 80, fut une découverte majeure qui révéla le rôle endocrinien du cœur. Les connaissances sur la relaxation vasculaire, la diurèse et la natriurèse provoquées par ce système ont évolué vers un niveau de complexité insoupçonné à cette époque. Nous savons à présent que les NP sont impliqués dans plusieurs autres mécanismes dont la prolifération cellulaire, l’apoptose, l’inhibition du système rénine-angiotensine-aldostérone (RAAS) et le métabolisme des adipocytes. Le métabolisme des lipides est maintenant devenu une cible de choix dans la lutte contre l’obésité. Cette condition aux proportions pandémiques est un facteur de risque majeur dans l’apparition de l’hypertension et du syndrome métabolique (MetS). La compréhension des mécanismes et des défauts de la voie des NP pourrait avoir un impact positif sur le contrôle du MetS et de l’hypertension. L’expression du récepteur des peptides natriuretiques de type 1 (NPR1/GCA) est contrôlée par plusieurs agents incluant son propre ligand, le peptide natriurétique de l’oreillette (ANP). La découverte d’une boucle de retro-inhibition, dans les années 90, a été un événement majeur dans le domaine des NP. En effet, suite à une stimulation à l’ANP, le NPR1/GCA peut inhiber l’activité transcriptionnelle de son propre gène par un mécanisme dépendant du cGMP. Notre groupe a identifié un élément cis-régulateur responsable de cette sensibilité au cGMP et mon projet consistait à identifier la ou les protéine(s) liant cet élément de réponse au cGMP (cGMP-RE). Nous avons identifié un clone liant le cGMP-RE en utilisant la technique du simple hybride chez la levure et une banque d’ADN complémentaire (ADNc) de rein humain. Ce clone provient d’un ADNc de 1083-bp dont le gène est localisé sur le chromosome 1 humain (1p33.36) et codant pour une protéine dont la fonction était inconnue jusqu’ici. Nous avons nommé cette nouvelle protéine GREBP en raison de sa fonction de cGMP Response Element Binding Protein. Des essais de liaison à l’ADN ont montré que cette protéine possède une affinité 18 fois plus élevée pour le cGMP-RE que le contrôle, tandis que des expériences de retard sur gel (EMSA) ont confirmé la spécificité des interactions protéine-ADN. De plus, l’immuno-précipitation de la chromatine (ChIP) a prouvé que GREBP lie le cGMP-RE dans des conditions physiologiques. La liaison de GREBP au cGMP-RE inhibe l’expression du gène rapporteur luciférase sous contrôle du promoteur de npr1/gca. L’inhibition de GREBP à l’aide d’ARN interférant active le promoteur de npr1/gca. Dans les cellules NCI-H295R, l’ANP stimule l’expression de grebp de 60% après seulement 3 heures et inhibe l’expression de npr1/gca de 30%. GREBP est une protéine nucléaire surtout exprimée dans le cœur et ayant le facteur eIF3F comme partenaire. Les variations nucléotidiques du gène sont plus fréquentes chez les patients hypertendus que chez des patients normotendus ou hypertendus souffrant de MetS. Nous rapportons ici l’existence d’un gène spécifique à l’humain qui agit comme répresseur transcriptionnel de npr1/gca et potentiellement impliqué dans le développement de l’hypertension.

Estudi dels carotenoides en espècies marrons de bacteris verds del sofre: diversitat, eco-fisiologia i regulació

El present treball es centra en l'estudi a diferents nivells dels carotenoides de les espècies marrons de Bacteris Verds del Sofre (GSB, de l'anglès Green Sulfur Bacteria). L'objectiu global ha estat el d'esbrinar quina és la funció d'aquests pigments dins l'aparell fotosintètic d'aquests microorganismes i aprofundir en el coneixement de la seva estructura i interaccions amb els altres pigments de l'aparell fotosintètic. En primer lloc es va dissenyar un nou mètode de cromatografia líquida d'alta resolució (HPLC) per analitzar de manera més ràpida i precisa els carotenoides de diferents soques de GSB (Capítol 3). Aquest mètode es basa en una purificació prèvia dels extractes pigmentaris amb columnes d'alúmina per eliminar les bacterioclorofil·les (BCls). Això va permetre analitzar amb una elevada resolució i en tan sols 45 min de carrera cromatogràfica els diferents carotenoides i els seus precursors, així com les configuracions trans i cis dels seus isòmers. El segon mètode utilitzat va consistir en una modificació del mètode de Borrego i Garcia-Gil (1994) i va permetre la separació precisa de tot tipus de pigments, procedents tant de cultius purs com de mostres de caràcter complex. Un exemple concret foren uns paleosediments de la zona lacustre de Banyoles. En aquests sediments (0,7-1,5 milions d'anys d'antiguitat) es van detectar, entre d'altres pigments, carotenoides específics de les espècies marrons de GSB, la qual cosa va permetre confirmar la presència d'aquests bacteris a la zona lacustre de Banyoles ja des del Pleistocè inferior. En aquest primer capítol també es van analitzar els carotenoides de Chlorobium (Chl.) phaeobacteroides CL1401 mitjançant cromatografia líquida acoblada a espectrometria de masses (LC-MS/MS), amb l'objectiu de confirmar la seva identificació i el seu pes molecular. A més, també es va avaluar l'efecte de la temperatura, la llum i diferents agents oxidants i reductors en la composició quantitativa i qualitativa dels carotenoides i les BCls d'aquesta espècie. Això va permetre confirmar el caràcter fotosensible de les BCls i que els isòmers trans/cis dels diferents carotenoides no són artefactes produïts durant la manipulació de les mostres, sinó que són constitutius de l'aparell fotosintètic d'aquests microorganismes. El Capítol 4 inclou els experiments de fisiologia duts a terme amb algunes espècies de GSB, a partir dels quals es va intentar esbrinar la dinàmica de síntesi dels diferents pigments de l'aparell fotosintètic (BCl antena, BCl a i carotenoides) durant el creixement d'aquestes espècies. Aquestes investigacions van permetre monitoritzar també els canvis en el nombre de centres de reacció (CR) durant el procés d'adaptació lumínica. La determinació experimental del nombre de CR es va realitzar a partir de la quantificació de la BCl663, l'acceptor primari en la cadena de transport d'electrons dels GSB. L'estimació del nombre de CR/clorosoma es va realitzar tant a partir de dades estequiomètriques i biomètriques presents a la bibliografia, com a partir de les dades experimentals obtingudes en el present treball. El bon ajust obtingut entre les diferents estimacions va donar solidesa al valor estequiomètric calculat, que fou, com a promig, d'uns 70 CR per clorosoma. En aquest capítol de fisiologia també es van estudiar les variacions en les relacions trans/cis pels principals carotenoides de les espècies marrons de GSB. Aquestes es van determinar a partir de cultius purs de laboratori i de poblacions naturals de GSB. Pel que fa als valors trobats en cultius de laboratori no es van observar diferències destacades entre el valor calculat a alta intensitat de llum i el calculat a baixa intensitat, essent en ambdós casos proper a 2. En els clorosomes aïllats de diferents soques marrons aquest quocient prengué un valor similar tant pels isòmers de l'isorenieratè (Isr) com pels del -isorenieratè (-Isr). En poblacions naturals de Chl. phaeobacteroides aquesta relació va ser també de 2 isòmers trans per cada isòmer cis, mantenint-se constant tant en fondària com al llarg del temps. Finalment, en el Capítol 5 es presenta un marcador molecular que permet la identificació específica d'espècies marrons de GSB. Malgrat que inicialment aquest marcador fou dissenyat a partir d'un gen implicat en la síntesi de carotenoides (crtY, el qual codifica per a una licopè ciclasa) la seqüència final a partir de la qual s'han aconseguit els encebadors selectius està relacionada amb la família de proteïnes de les Policètid-ceto-sintases (PKT). Tot i així, l'eina dissenyada pot ser de gran utilitat per a la discriminació d'espècies marrons de GSB respecte les verdes en poblacions mixtes com les que es troben en ambients naturals i obre la porta a futurs experiments d'ecologia microbiana utilitzant tècniques com la PCR en temps real, que permetria la monitorització selectiva de les poblacions d'espècies marrons de GSB en ecosistemes naturals.

Nitrosopersulfide (SSNO-) targets soluble guanylyl cyclase and induces vasodilation in vivo

Background Recent experimental evidence suggests that nitric oxide (NO) and hydrogen sulfide signaling pathways are intimately intertwined particularly in the vasculature, with mutual attenuation or potentiation of biological responses under control of the soluble guanylyl cyclase (sGC) / phopshodiesterase (PDE) pathway. There is now compelling evidence that part of the NO/sulfide cross talk has a chemical foundation via the formation of S/N-hybrid molecules including thionitrous acid (HSNO) and nitrosopersulfde (SSNO-). The aim of this study was to characterize the bioactive products of the interaction between sulfide and NO metabolites targeting sGC that may potentially regulate vasodilation. Results We found that the chemical interaction of sulfide with NO or nitrosothiols leads to formation of S/N-hybrid metabolites including SSNO- via intermediate formation of HSNO. Contrary to a recent report in the literature but consistent with the transient nature of HSNO, its formation was not detectable by high-resolution mass spectrometry under physiologically relevant conditions. SSNO- is also formed in non-aqueous media by the reaction of nitrite with oxidized sulfur species including colloidal sulfur and polysulfides. SSNO- is stable in the presence of high concentrations of thiols, release NO, and activates sGC in RFL-6 cells in an NO-dependent fashion. Moreover, SSNO- is a potent vasodilator in aortic rings in vitro and lowers blood pressure in rats in vivo. The presence of high concentrations of SOD or thiols does not affect SSNO- mediated sGC activation, while it potentiates and inhibits the effects of the nitroxyl (HNO) donor Angeli's salt, suggesting that HNO release from SSNO- is not involved in sGC activation. Conclusion The reaction between NO and sulfide leads to fomation of S/N-hybrid molecules including SSNO-, releasing NO, activating sGC and inducing vasodilation. SSNO- is considerably more stable than HSNO at pH 7.4 and thus a more likely biological mediator that can account for the chemical cross-talk between NO and sulfide.

Comparative effects of acute and subacute lycopene administration on chromosomal aberrations induced by cisplatin in male rats

Lycopene is a natural carotenoid, free radical scavenger, and presents protective effects by inhibiting oxidative DNA damage. The objective of the current study was to investigate the cytogenetic effects of a single acute and four daily gavage administrations of lycopene, and to examine possible protective effects on chromosomal damage induced by the antitumor drug cisplatin (cDDP) in rat bone marrow cells. The animals were divided into treatment groups, with three lycopene doses in the acute treatment (2, 4, and 6 mg/kg b.w.), three lycopene doses in the subacute treatment (0.5, 1.0, and 1.5 mg/kg b.w.) with and without cDDP (5 mg/kg b.w. i.p.), and respective controls. The results indicated that lycopene is neither cytotoxic nor clastogenic when compared with the negative controls (P > 0.01). cDDP-treated animals submitted to acute and subacute treatments with different lycopene doses showed a significant reduction (p < 0.01) in the number of abnormal metaphases when compared with the animals treated only with cDDP. The protective effects of lycopene on cDDP-induced chromosomal damage may be attributed to its antioxidant activity. These results suggest that this carotenoid may prove useful in reducing some of the toxic effects associated with certain classes of chemotherapeutic agents. (c) 2006 Elsevier Ltd. All rights reserved.

Antigenotoxicity and antimutagenicity of lycopene in HepG2 cell line evaluated by the comet assay and micronucleus test

Epidemiological studies have provided evidence that high consumption of tomatoes effectively reduces the risk of reactive oxygen species (ROS)-mediated diseases such as cancer. Tomatoes are rich sources of lycopene, a potent singlet oxygen-quenching carotenoid. In addition to its antioxidant properties, lycopene shows an array of biological effects including antimutagenic and anticarcinogenic activities. In the present study, the chemopreventive action of lycopene was examined on DNA damage and clastogenic or aneugenic effects of H2O2 and n-nitrosodiethylamine (DEN) in the metabolically competent human hepatoma cell line (HepG2 cells). Lycopene at concentrations of 10. 25, and 50 mu M, was tested under three protocols: before, simultaneously, and after treatment with the mutagen, using the comet and micronucleus assays. Lycopene significantly reduced the genotoxicity and mutagenicity of H2O2 in all of the conditions tested. For DEN, significant reductions of primary DNA damage (comet assay) were detected when the carotenoid (all of the doses) was added in the cell culture medium before or simultaneously with the mutagen. In the micronucleus test, the protective effect of lycopene was observed only when added prior to DEN treatment. In conclusion, our results suggest that lycopene is a suitable agent for preventing chemically-induced DNA and chromosome damage. (C) 2007 Elsevier Ltd. All rights reserved.