960 resultados para Biophysical requalification


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Le Programme des aides familiales résidantes (PAFR) a été en vigueur de 1992 jusqu’au 30 novembre 2014. Il s’agit d’un programme fédéral canadien par lequel des femmes originaires des Philippines en majorité et ayant obtenu un diplôme d’études postsecondaires migraient partout au Canada. Sous ce programme, celles-ci se retrouvaient avec un statut de migrante temporaire. Elles avaient la possibilité d’obtenir leur résidence permanente après avoir complété 24 mois de travail en tant qu’aides familiales. Elles devaient travailler au domicile privé d’un employeur dont le nom était inscrit sur leur permis de travail, chez qui elles avaient aussi l’obligation de résidence. En travaillant en tant qu’aides familiales résidantes, ces femmes étaient et sont toujours placées dans une situation de déqualification professionnelle inévitable qui semble se prolonger à long terme. Cette situation est notamment attribuable à la non-reconnaissance de leurs qualifications étrangères ainsi qu’à la structure du PAFR. Cette recherche porte donc sur le parcours de requalification professionnelle de ces migrantes, que certaines utilisent comme stratégie pour remédier à cette situation. En plus de s’intéresser à leur période de travail en tant qu’aides familiales résidantes, cette recherche étudie le parcours de requalification professionnelle en quatre étapes : la décision de participer à la formation ; le choix de l’orientation professionnelle ; le déroulement des études ; l’accès à un emploi correspondant à la formation. L’objectif est de connaître l’impact qu’aura le passage de celles-ci par le PAFR, sur leur parcours de requalification. Les résultats obtenus à l’aide de onze entretiens semi-dirigés, avec des femmes étant venues à Montréal par le programme, montrent qu’à long terme, le diplôme obtenu localement peut améliorer une situation professionnelle, mais ce n’est pas une garantie. Des obstacles comme l’isolement ainsi que l’impossibilité pour ces femmes de se construire un capital financier et social persistent et affectent toutes celles ayant migré sous le programme, qu’elles aient fait un retour aux études ou non.

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Le Programme des aides familiales résidantes (PAFR) a été en vigueur de 1992 jusqu’au 30 novembre 2014. Il s’agit d’un programme fédéral canadien par lequel des femmes originaires des Philippines en majorité et ayant obtenu un diplôme d’études postsecondaires migraient partout au Canada. Sous ce programme, celles-ci se retrouvaient avec un statut de migrante temporaire. Elles avaient la possibilité d’obtenir leur résidence permanente après avoir complété 24 mois de travail en tant qu’aides familiales. Elles devaient travailler au domicile privé d’un employeur dont le nom était inscrit sur leur permis de travail, chez qui elles avaient aussi l’obligation de résidence. En travaillant en tant qu’aides familiales résidantes, ces femmes étaient et sont toujours placées dans une situation de déqualification professionnelle inévitable qui semble se prolonger à long terme. Cette situation est notamment attribuable à la non-reconnaissance de leurs qualifications étrangères ainsi qu’à la structure du PAFR. Cette recherche porte donc sur le parcours de requalification professionnelle de ces migrantes, que certaines utilisent comme stratégie pour remédier à cette situation. En plus de s’intéresser à leur période de travail en tant qu’aides familiales résidantes, cette recherche étudie le parcours de requalification professionnelle en quatre étapes : la décision de participer à la formation ; le choix de l’orientation professionnelle ; le déroulement des études ; l’accès à un emploi correspondant à la formation. L’objectif est de connaître l’impact qu’aura le passage de celles-ci par le PAFR, sur leur parcours de requalification. Les résultats obtenus à l’aide de onze entretiens semi-dirigés, avec des femmes étant venues à Montréal par le programme, montrent qu’à long terme, le diplôme obtenu localement peut améliorer une situation professionnelle, mais ce n’est pas une garantie. Des obstacles comme l’isolement ainsi que l’impossibilité pour ces femmes de se construire un capital financier et social persistent et affectent toutes celles ayant migré sous le programme, qu’elles aient fait un retour aux études ou non.

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The Jaíba Irrigated Perimeter is a large irrigated agriculturearea, located in the region Forest Jaíba between the SãoFrancisco and Verde Grande rivers, in the Brazilian semi-arid region. In 2014, irrigators thisthe region face losses in theinterruption of new plantings in irrigated areas due to water scarcity. The objective ofthis study is combine the modelto estimate the Monteith BIO with the SAFER algorithm in the case of obtaining ET, to analyze the dynamics of naturalvegetation and irrigated crops in water scarcity period. For application of the model are necessary data frommeteorological stations and satellite images. Were used 23satellite images of MODIS withspatial resolution of 250mand temporal 16 days, of 2014 year. For analyze the results,we used central pivots irrigation mask of Minas Geraisstate, Brazil. In areas with irrigated agriculture with central pivot, the mean values of BIO over the year 2014 were88.96 kg.ha-1.d-1. The highest values occurred between April 23 and May 8, with BIO 139 kg.ha-1.d-1. For areas withnatural vegetation, the average BIO was 88.34 kg.ha-1.d-1with lower values in September. Estimates of ET varied withthe lowest values of ET observedin natural vegetation 1,91±1,22 mm.d-1and the highest values in irrigated area isobserved 3,51±0,97 mm.d-1. Results of this study can assist in monitoring of river basins, contributing to themanagement irrigated agriculture, with the trend of scarcity of water resources and increasing conflicts for the wateruse.

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Canopy and aerodynamic conductances (gC and gA) are two of the key land surface biophysical variables that control the land surface response of land surface schemes in climate models. Their representation is crucial for predicting transpiration (λET) and evaporation (λEE) flux components of the terrestrial latent heat flux (λE), which has important implications for global climate change and water resource management. By physical integration of radiometric surface temperature (TR) into an integrated framework of the Penman?Monteith and Shuttleworth?Wallace models, we present a novel approach to directly quantify the canopy-scale biophysical controls on λET and λEE over multiple plant functional types (PFTs) in the Amazon Basin. Combining data from six LBA (Large-scale Biosphere-Atmosphere Experiment in Amazonia) eddy covariance tower sites and a TR-driven physically based modeling approach, we identified the canopy-scale feedback-response mechanism between gC, λET, and atmospheric vapor pressure deficit (DA), without using any leaf-scale empirical parameterizations for the modeling. The TR-based model shows minor biophysical control on λET during the wet (rainy) seasons where λET becomes predominantly radiation driven and net radiation (RN) determines 75 to 80 % of the variances of λET. However, biophysical control on λET is dramatically increased during the dry seasons, and particularly the 2005 drought year, explaining 50 to 65 % of the variances of λET, and indicates λET to be substantially soil moisture driven during the rainfall deficit phase. Despite substantial differences in gA between forests and pastures, very similar canopy?atmosphere "coupling" was found in these two biomes due to soil moisture-induced decrease in gC in the pasture. This revealed the pragmatic aspect of the TR-driven model behavior that exhibits a high sensitivity of gC to per unit change in wetness as opposed to gA that is marginally sensitive to surface wetness variability. Our results reveal the occurrence of a significant hysteresis between λET and gC during the dry season for the pasture sites, which is attributed to relatively low soil water availability as compared to the rainforests, likely due to differences in rooting depth between the two systems. Evaporation was significantly influenced by gA for all the PFTs and across all wetness conditions. Our analytical framework logically captures the responses of gC and gA to changes in atmospheric radiation, DA, and surface radiometric temperature, and thus appears to be promising for the improvement of existing land?surface?atmosphere exchange parameterizations across a range of spatial scales.

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Canopy and aerodynamic conductances (gC and gA) are two of the key land surface biophysical variables that control the land surface response of land surface schemes in climate models. Their representation is crucial for predicting transpiration (?ET) and evaporation (?EE) flux components of the terrestrial latent heat flux (?E), which has important implications for global climate change and water resource management. By physical integration of radiometric surface temperature (TR) into an integrated framework of the Penman?Monteith and Shuttleworth?Wallace models, we present a novel approach to directly quantify the canopy-scale biophysical controls on ?ET and ?EE over multiple plant functional types (PFTs) in the Amazon Basin. Combining data from six LBA (Large-scale Biosphere-Atmosphere Experiment in Amazonia) eddy covariance tower sites and a TR-driven physically based modeling approach, we identified the canopy-scale feedback-response mechanism between gC, ?ET, and atmospheric vapor pressure deficit (DA), without using any leaf-scale empirical parameterizations for the modeling. The TR-based model shows minor biophysical control on ?ET during the wet (rainy) seasons where ?ET becomes predominantly radiation driven and net radiation (RN) determines 75 to 80?% of the variances of ?ET. However, biophysical control on ?ET is dramatically increased during the dry seasons, and particularly the 2005 drought year, explaining 50 to 65?% of the variances of ?ET, and indicates ?ET to be substantially soil moisture driven during the rainfall deficit phase. Despite substantial differences in gA between forests and pastures, very similar canopy?atmosphere "coupling" was found in these two biomes due to soil moisture-induced decrease in gC in the pasture. This revealed the pragmatic aspect of the TR-driven model behavior that exhibits a high sensitivity of gC to per unit change in wetness as opposed to gA that is marginally sensitive to surface wetness variability. Our results reveal the occurrence of a significant hysteresis between ?ET and gC during the dry season for the pasture sites, which is attributed to relatively low soil water availability as compared to the rainforests, likely due to differences in rooting depth between the two systems. Evaporation was significantly influenced by gA for all the PFTs and across all wetness conditions. Our analytical framework logically captures the responses of gC and gA to changes in atmospheric radiation, DA, and surface radiometric temperature, and thus appears to be promising for the improvement of existing land?surface?atmosphere exchange parameterizations across a range of spatial scales.

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In prokaryotic organisms, lower eukaryotes and plants, some important biological reactions are catalyzed by nickel-dependent enzymes, making this metal ion essential microelement for their life. On the other hand, excessive concentration of nickel into the cell, or prolonged exposure to nickel compounds, has toxic effects in living organisms. In addition, nickel has been classified by IARC as Group I human carcinogen, because of the correlation between its inhalation and increased incidence of nasal and lung cancers. The aim of this work was to investigate the nickel impact on human health, considering both its direct role on human cells and its indirect effect as essential element for human important bacteria. In humans, nickel induces N-myc downstream regulated gene 1 (NDRG1) expression, recently proposed as new target in cancer therapy. CD, light scattering and ITC were applied on the recombinant full-length protein and its C-terminal intrinsically disordered domain, for studying the NDRG1 structural and functional properties. In particular, the fold and dynamics of the C-terminal region were examined by NMR spectroscopy and site-directed spin labeling coupled to EPR, showing the features of an intrinsically disordered region. In nickel-dependent bacteria, nickel metabolism is strictly regulated, through the activity of different transcription factors. In Streptomyces griseus the expression of two superoxide dismutases (SODs) is antagonistically regulated by nickel thanks to the transcriptional complex SgSrnR/SgSrnQ. The SgSrnR protein was heterologously expressed and its activity as possible nickel sensor studied. DNaseI footprinting and β-galactosidase gene reporter assays revealed that SgSrnR functions as transcriptional activator, prompting the hypothesis of a new model to describe the activity of this complex. In addition, ITC, NMR and X-ray crystallography demonstrated that SgSrnR presents the fold typical of ArsR/SmtB transcription factors and low metal binding affinity, non compatible with a role as a nickel-sensor, function probably played by its partner SgSrnQ.

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RAD52 is a protein involved in various DNA reparation mechanisms. In the last few years, RAD52 has been proposed as a novel pharmacological target for cancer synthetic lethality strategies. Hence, this work has the purpose to investigate RAD52 protein, with biophysical and structural tools to shed light on proteins features and mechanistic details that are, up to now poorly described, and to design novel strategies for its inhibition. My PhD work had two goals: the structural and functional characterization of RAD52 and the identification of novel RAD52 inhibitors. For the first part, RAD52 was characterized both for its DNA interaction and oligomerization state together with its propensity to form high molecular weight superstructures. Moreover, using EM and Cryo-EM techniques, additional RAD52 structural hallmarks were obtained, valuable both for understanding protein mechanism of action and for drug discovery purpose. The second part of my PhD project focused on the design and characterization of novel RAD52 inhibitors to be potentially used in combination therapies with PARPi to achieve cancer cells synthetic lethality, avoiding resistance occurrence and side effects. With this aim we selected and characterized promising RAD52 inhibitors through three different approaches: 19F NMR fragment-based screening; virtual screening campaign; aptamers computational design. Selected hits (fragments, molecules and aptamers) were investigated for their binding to RAD52 and for their mechanism of inhibition. Collected data highlighted the identification of hits worthy to be developed into more potent and selective RAD52 inhibitors. Finally, a side project carried out during my PhD is reported. GSK-3β protein, an already validated pharmacological target was investigated using biophysical and structural biology tools. Here, an innovative and adaptable drug discovery screening pipeline able to directly identify selective compounds with binding affinities not higher than a reference binder was developed.

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Recent experiments have revealed the fundamental importance of neuromodulatory action on activity-dependent synaptic plasticity underlying behavioral learning and spatial memory formation. Neuromodulators affect synaptic plasticity through the modification of the dynamics of receptors on the synaptic membrane. However, chemical substances other than neuromodulators, such as receptors co-agonists, can influence the receptors' dynamics and thus participate in determining plasticity. Here we focus on D-serine, which has been observed to affect the activity thresholds of synaptic plasticity by co-activating NMDA receptors. We use a computational model for spatial value learning with plasticity between two place cell layers. The D-serine release is CB1R mediated and the model reproduces the impairment of spatial memory due to the astrocytic CB1R knockout for a mouse navigating in the Morris water maze. The addition of path-constraining obstacles shows how performance impairment depends on the environment's topology. The model can explain the experimental evidence and produce useful testable predictions to increase our understanding of the complex mechanisms underlying learning.

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Il lavoro di questa tesi ha riguardato lo studio e l'analisi acustica del sito della Scuola di Ingegneria di Bologna in Via Terracini con l’obiettivo di trovare una soluzione per mitigare il disturbo sonoro rilevato nella zona. Dalle diverse misurazioni fonometriche si è stabilita l’entità numerica di partenza che ha richiesto un’implentazione attraverso delle misurazioni di tipo qualitativo con delle registrazioni in formato wave ed annotando al contempo il numero ed il tipo di veicoli che circolavano. Attraverso l'implementazione dei vari dati ricavati dai rilievi delle diverse sezioni, è stata convalidata la consapevolezza del principale problema dell'area: l'inquinamento acustico causato dal traffico stradale. Lo studio è proseguito monitorando le bande di frequenza che interessavano questo tipo di rumore e quantificando il livello in dB che superava i limiti imposti dalla normativa. Non essendo possibile ridurre il traffico acustico direttamente alla fonte, si è optato per misure di protezione del recettore che si sono configurate nell'inserimento dell'asfalto fonoassorbente e nella progettazione di una barriera acustica. Dopo aver ricercato i diversi tipi di barriere, si è deciso per la barriera con cristalli sonici. Si è indagato quali fattori influissero sulle proprietà dei cristalli sonici: la distribuzione geometrica, la frazione di riempimento e la dimensione degli elementi periodici influenzano la formazione delle bande proibite, la loro larghezza e quindi la capacità fonoisolante del cristallo sonico. In base all'altezza degli edifici e alla loro distanza dalla strada, Via Terracini è stata suddivisa in quattro sezioni. Infine, per le varie sezioni, è stato riprodotto il grafico frequenza/perdita di inserzione che ha confermato una riduzione del livello sonoro di quasi 15 dB. Il progetto ha previsto anche il dimensionamento e la verifica delle azioni statiche (carichi e azione del vento) e dell'azione sismica del sistema strutturale della barriera.

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The raft hypothesis proposes that microdomains enriched in sphingolipids, cholesterol, and specific proteins are transiently formed to accomplish important cellular tasks. Equivocally, detergent-resistant membranes were initially assumed to be identical to membrane rafts, because of similarities between their compositions. In fact, the impact of detergents in membrane organization is still controversial. Here, we use phase contrast and fluorescence microscopy to observe giant unilamellar vesicles (GUVs) made of erythrocyte membrane lipids (erythro-GUVs) when exposed to the detergent Triton X-100 (TX-100). We clearly show that TX-100 has a restructuring action on biomembranes. Contact with TX-100 readily induces domain formation on the previously homogeneous membrane of erythro-GUVs at physiological and room temperatures. The shape and dynamics of the formed domains point to liquid-ordered/liquid-disordered (Lo/Ld) phase separation, typically found in raft-like ternary lipid mixtures. The Ld domains are then separated from the original vesicle and completely solubilized by TX-100. The insoluble vesicle left, in the Lo phase, represents around 2/3 of the original vesicle surface at room temperature and decreases to almost 1/2 at physiological temperature. This chain of events could be entirely reproduced with biomimetic GUVs of a simple ternary lipid mixture, 2:1:2 POPC/SM/chol (phosphatidylcholine/sphyngomyelin/cholesterol), showing that this behavior will arise because of fundamental physicochemical properties of simple lipid mixtures. This work provides direct visualization of TX-100-induced domain formation followed by selective (Ld phase) solubilization in a model system with a complex biological lipid composition.

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Hsp90 is a molecular chaperone essential for cell viability in eukaryotes that is associated with the maturation of proteins involved in important cell functions and implicated in the stabilization of the tumor phenotype of various cancers, making this chaperone a notably interesting therapeutic target. Celastrol is a plant-derived pentacyclic triterpenoid compound with potent antioxidant, anti-inflammatory and anticancer activities; however, celastrol's action mode is still elusive. In this work, we investigated the effect of celastrol on the conformational and functional aspects of Hsp90α. Interestingly, celastrol appeared to target Hsp90α directly as the compound induced the oligomerization of the chaperone via the C-terminal domain as demonstrated by experiments using a deletion mutant. The nature of the oligomers was investigated by biophysical tools demonstrating that a two-fold excess of celastrol induced the formation of a decameric Hsp90α bound throughout the C-terminal domain. When bound, celastrol destabilized the C-terminal domain. Surprisingly, standard chaperone functional investigations demonstrated that neither the in vitro chaperone activity of protecting against aggregation nor the ability to bind a TPR co-chaperone, which binds to the C-terminus of Hsp90α, were affected by celastrol. Celastrol interferes with specific biological functions of Hsp90α. Our results suggest a model in which celastrol binds directly to the C-terminal domain of Hsp90α causing oligomerization. However, the ability to protect against protein aggregation (supported by our results) and to bind to TPR co-chaperones are not affected by celastrol. Therefore celastrol may act primarily by inducing specific oligomerization that affects some, but not all, of the functions of Hsp90α. To the best of our knowledge, this study is the first work to use multiple probes to investigate the effect that celastrol has on the stability and oligomerization of Hsp90α and on the binding of this chaperone to Tom70. This work provides a novel mechanism by which celastrol binds Hsp90α.

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Remote sensing data are each time more available and can be used to monitor the vegetal development of main agricultural crops, such as the Arabic coffee in Brazil, since that the relationship between spectral and agronomical data be well known. Therefore, this work had the main objective to assess the use of Quickbird satellite images to estimate biophysical parameters of coffee crop. Test area was composed by 25 coffee fields located between the cities of Ribeirão Corrente, Franca and Cristais Paulista (SP), Brazil, and the biophysical parameters used were row and between plants spacing, plant height, LAI, canopy diameter, percentage of vegetation cover, roughness and biomass. Spectral data were the reflectance of four bands of QUICKBIRD and values of four vegetations indexes (NDVI, GVI, SAVI and RVI) based on the same satellite. All these data were analyzed using linear and nonlinear regression methods to generate estimation models of biophysical parameters. The use of regression models based on nonlinear equations was more appropriate to estimate parameters such as the LAI and the percentage of biomass, important to indicate the productivity of coffee crop.

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For the diagnosis and prognosis of the problems of quality of life, a multidisciplinary ecosystemic approach encompasses four dimensions of being-in-the-world, as donors and recipients: intimate, interactive, social and biophysical. Social, cultural and environmental vulnerabilities are understood and dealt with, in different circumstances of space and time, as the conjugated effect of all dimensions of being-in-the-world, as they induce the events (deficits and assets), cope with consequences (desired or undesired) and contribute for change. Instead of fragmented and reduced representations of reality, diagnosis and prognosis of cultural, educational, environmental and health problems considers the connections (assets) and ruptures (deficits) between the different dimensions, providing a planning model to develop and evaluate research, teaching programmes, public policies and field projects. The methodology is participatory, experiential and reflexive; heuristic-hermeneutic processes unveil cultural and epistemic paradigms that orient subject-object relationships; giving people the opportunity to reflect on their own realities, engage in new experiences and find new ways to live better in a better world. The proposal is a creative model for thought and practice, providing many opportunities for discussion, debate and development of holistic projects integrating different scientific domains (social sciences, psychology, education, philosophy, etc.)

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Interleukin-22 (IL-22) is a class 2 cytokine whose primary structure is similar to that of interleukin 10 (IL-10) and interferon-gamma (IFN-gamma). IL-22 induction during acute phase immune response indicates its involvement in mechanisms of inflammation. Structurally different from IL-10 and a number of other members of IL-10 family, which form intertwined inseparable V-shaped dimers of two identical polypeptide chains, a single polypeptide chain of IL-22 folds on itself in a relatively globular structure. Here we present evidence, based on native gel electrophoresis, glutaraldehyde cross-linking, dynamic light scattering, and small angle x-ray scattering experiments, that human IL-22 forms dimers and tetramers in solution under protein concentrations assessable by these experiments. Unexpectedly, low-resolution molecular shape of IL-22 dimers is strikingly similar to that of IL-10 and other intertwined cytokine dimeric forms. Furthermore, we determine an ab initio molecular shape of the IL-22/IL-22R1 complex which reveals the V-shaped IL-22 dimer interacting with two cognate IL-22R1 molecules. Based on this collective evidence, we argue that dimerization might be a common mechanism of all class 2 cytokines for the molecular recognition with their respective membrane receptor. We also speculate that the IL-22 tetramer formation could represent a way to store the cytokine in nonactive form at high concentrations that could be readily converted into functionally active monomers and dimers upon interaction with the cognate cellular receptors.

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Dihydroorotate dehydrogenase (DHODH) catalyzes the oxidation of dihydroorotate to orotate during the fourth step of the de novo pyrimidine synthesis pathway. In rapidly proliferating mammalian cells, pyrimidine salvage pathway is insufficient to overcome deficiencies in that pathway for nucleotide synthesis. Moreover, as certain parasites lack salvage enzymes, relying solely on the de novo pathway, DHODH inhibition has turned out as an efficient way to block pyrimidine biosynthesis. Escherichia coli DHODH (EcDHODH) is a class 2 DHODH, found associated to cytosolic membranes through an N-terminal extension. We used electronic spin resonance (ESR) to study the interaction of EcDHODH with vesicles of 1,2-dioleoyl-sn-glycero-phosphatidylcholine/detergent. Changes in vesicle dynamic structure induced by the enzyme were monitored via spin labels located at different positions of phospholipid derivatives. Two-component ESR spectra are obtained for labels 5- and 1 0-phosphatidylcholine in presence of EcDHODH, whereas other probes show a single-component spectrum. The appearance of an additional spectral component with features related to fast-motion regime of the probe is attributed to the formation of a defect-like structure in the membrane hydrophobic region. This is probably the mechanism used by the protein to capture quinones used as electron acceptors during catalysis. The use of specific spectral simulation routines allows us to characterize the ESR spectra in terms of changes in polarity and mobility around the spin-labeled phospholipids. We believe this is the first report of direct evidences concerning the binding of class 2 DHODH to membrane systems.