Pruebas preliminares sobre toxicidad aguda del cobre en la Concha de Abanico (Argopecten purpuratus)

Se realizaron siete bioensayos estáticos con la Concha de Abanico (Argopecten purpuratus) con concentraciones de cobre que variaron de 0.007 a 0.74 ppm. El agua de mar de donde provinieron los individuos usados en el experimento tenía concentraciones de cobre entre 0.005-0.007 ppm. La bioacumulación inicial en los animales varó de 1.69 a 6.50 ppm. Como resultado preliminar se determinó que 0.13 ppm es la concentración letal media (LC 50) en 96 horas.

In vivo characterization of sunitinib eluting beads

Purpose: To study the pharmacokinetics and potential toxicity of sunitinib eluting beads in an animal modelMaterials: Healthy New-Zealand white rabbits were used. 8 animals received 0.2ml of DC Beads loaded with 6mg of sunitinb intra arterially in the hepatic artery (group 1) and 4 animals received 6mg of sunitinib administered orally (group 2). In group 1, animals were sacrificed 6 hours (n=4) and 24 hours (n=4) after embolization. In group 2, animals were sacrificed 6 hours (n=2) and 24 hours (n=2) after oral administration of sunitinib. Liver enzymes were measured at 0, 6 and 24 hours in both groups. Plasmatic sunitinib concentration was measured by LC MS/MS tandem mass spectroscopy at 0, 1, 2, 3, 4, 5, 6 and 24 hours. At sacrifice, the livers were harvested and sunitinib concentration in liver tissue was assessed by LC MS/MS tandem mass spectroscopy.Results: After embolization we observed an expected elevation of AST and ALT. Serial plasmatic measurments after embolization showed a very low sunitinib concentration (<50ng/ml). Measurment of sunitinib in the embolized liver tissue showed a very high concentration at 6 hours (3870ng/ml) and 24 hours (4741.7ng/ml).Conclusions: Sunitinib eluting beads are well tolerated by rabbits when administered intra-arterially in the hepatic artery. No unexpected toxicity was observed. Very high drug concentration canbe obtained at the site of embolization with minimal systemic passage.

Food's visually perceived fat content affects discrimination speed in an orthogonal spatial task.

Choosing what to eat is a complex activity for humans. Determining a food's pleasantness requires us to combine information about what is available at a given time with knowledge of the food's palatability, texture, fat content, and other nutritional information. It has been suggested that humans may have an implicit knowledge of a food's fat content based on its appearance; Toepel et al. (Neuroimage 44:967-974, 2009) reported visual-evoked potential modulations after participants viewed images of high-energy, high-fat food (HF), as compared to viewing low-fat food (LF). In the present study, we investigated whether there are any immediate behavioural consequences of these modulations for human performance. HF, LF, or non-food (NF) images were used to exogenously direct participants' attention to either the left or the right. Next, participants made speeded elevation discrimination responses (up vs. down) to visual targets presented either above or below the midline (and at one of three stimulus onset asynchronies: 150, 300, or 450 ms). Participants responded significantly more rapidly following the presentation of a HF image than following the presentation of either LF or NF images, despite the fact that the identity of the images was entirely task-irrelevant. Similar results were found when comparing response speeds following images of high-carbohydrate (HC) food items to low-carbohydrate (LC) food items. These results support the view that people rapidly process (i.e. within a few hundred milliseconds) the fat/carbohydrate/energy value or, perhaps more generally, the pleasantness of food. Potentially as a result of HF/HC food items being more pleasant and thus having a higher incentive value, it seems as though seeing these foods results in a response readiness, or an overall alerting effect, in the human brain.

Quantitative monitoring of tamoxifen in human plasma extended to 40 metabolites using liquid-chromatography high-resolution mass spectrometry: new investigation capabilities for clinical pharmacology.

Liquid-chromatography (LC) high-resolution (HR) mass spectrometry (MS) analysis can record HR full scans, a technique of detection that shows comparable selectivity and sensitivity to ion transitions (SRM) performed with triple-quadrupole (TQ)-MS but that allows de facto determination of "all" ions including drug metabolites. This could be of potential utility in in vivo drug metabolism and pharmacovigilance studies in order to have a more comprehensive insight in drug biotransformation profile differences in patients. This simultaneous quantitative and qualitative (Quan/Qual) approach has been tested with 20 patients chronically treated with tamoxifen (TAM). The absolute quantification of TAM and three metabolites in plasma was realized using HR- and TQ-MS and compared. The same LC-HR-MS analysis allowed the identification and relative quantification of 37 additional TAM metabolites. A number of new metabolites were detected in patients' plasma including metabolites identified as didemethyl-trihydroxy-TAM-glucoside and didemethyl-tetrahydroxy-TAM-glucoside conjugates corresponding to TAM with six and seven biotransformation steps, respectively. Multivariate analysis allowed relevant patterns of metabolites and ratios to be associated with TAM administration and CYP2D6 genotype. Two hydroxylated metabolites, α-OH-TAM and 4'-OH-TAM, were newly identified as putative CYP2D6 substrates. The relative quantification was precise (<20 %), and the semiquantitative estimation suggests that metabolite levels are non-negligible. Metabolites could play an important role in drug toxicity, but their impact on drug-related side effects has been partially neglected due to the tremendous effort needed with previous MS technologies. Using present HR-MS, this situation should evolve with the straightforward determination of drug metabolites, enlarging the possibilities in studying inter- and intra-patients drug metabolism variability and related effects.

Compressibilidade de três Latossolos em função da umidade e uso

Por meio do ensaio de compressibilidade, foram estudados os efeitos do manejo e da umidade na pressão de preconsolidação (σp) de três solos: Latossolo Vermelho-Amarelo, Latossolo Roxo e Latossolo Vermelho-Escuro sob cultura anual, mata natural e pastagem, na região de Lavras (MG), em duas profundidades (0-0,03 e 0,27-0,30 m). Para cada condição, foram coletadas cinco amostras indeformadas e uma deformada, com três repetições. As amostras, indeformadas e com diferentes umidades, foram utilizadas no ensaio de compressão uniaxial, obtendo-se as curvas de compressão, das quais foram extraídas as respectivas pressões de preconsolidação. Com as amostras deformadas, determinaram-se os limites de plasticidade e de contração, textura e matéria orgânica. Os modelos de compactação testados foram baseados na pressão de preconsolidação e na umidade do solo. Para uma mesma condição e profundidade, houve diferença significativa entre os valores dos teores de argila e areia nos três solos. Os valores da densidade do solo inicial (Ds i) foram estatisticamente diferentes para todas as condições na mesma profundidade, exceto na camada de 0-0,03 m para a cultura anual. À medida que a umidade do solo aumentou, a pressão de preconsolidação decresceu exponencialmente, indicando uma redução na capacidade de suporte de carga do solo. O LR apresentou, em geral, maior capacidade de suporte de carga do que o LE e LV. Essa maior capacidade de suporte de carga pode estar associada com o seu maior teor de argila e menor teor de areia. A capacidade de suporte de carga na zona de friabilidade variou de 154 a 167 kPa, para o LV; de 77 a 183 kPa, para o LR, e de 77 a 132 kPa, para o LE. As umidades 0,33 a 0,30 kg kg-1, 0,42 a 0,27 kg kg-1, e 0,35 a 0,33 kg kg-1 correspondem à faixa de friabilidade (LP - LC) do LV, LR e LE, respectivamente. O modelo baseado na história de tensão do solo evidenciou o efeito da compactação causada pelas máquinas de preparo do solo na camada de 0,27-0,30 m, para a cultura anual, enquanto, para a pastagem, ocorreu o efeito do pisoteio do gado na camada de 0-0,03 m.

Aspectos biológico - pesqueros del Camaroncito Rojo Pleuroncodes monodon (M Edwards, 1837) en el litoral peruano

Se analizaron algunos parámetros poblacionales de 11 583 ejemplares del Pleuroncodes monodon (CRUSTACEA: ANOMURA: GALAYHEIDAE) distribuidos en 5 860 (52%) ejemplares machos y 5723 (48%) ejemplares hembras, las muestras se obtuvieron de los lances de comprobación de los cruceros de evaluación tanto para recursos pelágicos y demersales ejecutados por el Instituto del Mar del Perú (IMARPE) entre febrero y noviembre del 2001, la zona de estudio estuvo comprendida entre Puerto Pizarro (03°29,1’ S; 80°23,0’ W) hasta Los Palos (18°20,18’ S; 70°22,5’ W). La presencia del Pleuroncodes monodon respecto a su distribución batimétrica abarcó entre 1,5 y 300 m. De profundidad. La distribución total de tallas fluctuó entre 6,0 y 42,4 mm de longitud cefalotoráxica (Lc) y entre 6,0 y 41,1 mm de Lc para machos y hembras respectivamente. Por estaciones la estructura por tallas fue variable denotándose dos marcados grupos de talla, predominando un primer grupo los denominados ”juveniles” de (de 6 hasta 20 mm de Lc), sin embargo aquellos ejemplares con longitud cefalotoráxica menor a los 13 mm se registraron mayormente en las estaciones de verano y primavera, mientras un segundo grupo perteneciente a los de talla intermedia denominados “adultos jóvenes” registrados en todas las estaciones. Su distribución horizontal fue mayormente costera; aunque su presencia fue hasta más de las 50 millas náuticas (mn) de la costa; sin embargo los mayores registros de este recurso fue dentro de las 25 mn de la costa, en cuanto a los ejemplares “juveniles” su presencia fue marcada dentro de las 10 mn de la costa. La composición sexual de la población muestreada varió durante el periodo de estudio obteniéndose una razón promedio de 1,02 (machos/hembras). Además se encontraron hembras con huevos en todo el periodo de estudio, sin embargo los altos porcentajes de hembras ovígeras se registraron entre las estaciones de invierno (70%) y primavera (68%) del total. La talla de madurez sexual en las hembras fue de 18,8 mm Lc, mientras que el primer desove en 17,4 mm de Lc. Al registrarse hembras ovígeras en todo el año, existe un periodo anual de postura (liberación de los huevos eclosionados), detectándose éste en la estacón de primavera. La relación longitud cefalotoráxica vs Fecundidad, está determinada por la ecuación: NHT 0,1899 Lc2,9883 Además se analizaron los siguientes parámetros: largo cefalotoráxico, peso total, peso del abdomen, características de los huevos. Complementándose tales evaluaciones con determinaciones de áreas de distribución y concentración, así como la relación de los factores: abióticos (temperaturas, salinidad y oxígeno) y bióticos en relación a predadores y competidores dentro de la cadena trófica.

Therapeutic Drug Monitoring (TDM) of Imatinib: Effectiveness of Bayesian dose adjustment for CML patients enrolled in the Imatinib Concentration Monitoring (I-COME) study

Introduction: As imatinib pharmacokinetics are highly variable, plasma levels differ largely between patients under the same dosage. Retrospective studies in chronic myeloid leukemia (CML) patients showed significant correlations between low levels and suboptimal response, as well as between high levels and poor tolerability. Monitoring of trough plasma levels, targeting 1000 μg/L and above, is thus increasingly advised. Our study was launched to assess prospectively the clinical usefulness of systematic imatinib TDM in CML patients. This preliminary analysis addresses the appropriateness of the dosage adjustment approach applied in this study, which targets the recommended trough level and allows an interval of 4-24 h after last drug intake for blood sampling. Methods: Blood samples from the first 15 patients undergoing 1st TDM were obtained 1.5-25 h after last dose. Imatinib plasma levels were measured by LC-MS/MS and the concentrations were extrapolated to trough based on a Bayesian approach using a population pharmacokinetic model. Trough levels were predicted to differ significantly from the target in 12 patients (10 <750 μg/L; 2 >1500 μg/L along with poor tolerance) and individual dose adjustments were proposed. 8 patients underwent a 2nd TDM cycle. Trough levels of 1st and 2nd TDM were compared, the sample drawn 1.5 h after last dose (during distribution phase) was excluded from the analysis. Results: Individual dose adjustments were applied in 6 patients. Observed concentrations extrapolated to trough ranged from 360 to 1832 μg/L (median 725; mean 810, CV 52%) on 1st TDM and from 720 to 1187 μg/L (median 950; mean 940, CV 18%) on 2nd TDM cycle. Conclusions: These preliminary results suggest that TDM of imatinib using a Bayesian interpretation is able to target the recommended trough level of 1000 μg/L and to reduce the considerable differences in trough level exposure between patients (with CV decreasing from 52% to 18%). While this may simplify blood collection in daily practice, as samples do not have to be drawn exactly at trough, the largest possible interval to last drug intake yet remains preferable to avoid sampling during distribution phase leading to biased extrapolation. This encourages the evaluation of the clinical benefit of a routine TDM intervention in CML patients, which the randomized Swiss I-COME trial aims to.

Quantification of glucuronidated and sulfated steroids in human urine by ultra-high pressure liquid chromatography quadrupole time-of-flight mass spectrometry.

The urinary steroid profile is constituted by anabolic androgenic steroids, including testosterone and its relatives, that are extensively metabolized into phase II sulfated or glucuronidated steroids. The use of liquid chromatography coupled to mass spectrometry (LC-MS) is an issue for the direct analysis of conjugated steroids, which can be used as urinary markers of exogenous steroid administration in doping analysis, without hydrolysis of the conjugated moiety. In this study, a sensitive and selective ultra high-pressure liquid chromatography coupled to quadrupole time-of-flight mass spectrometer (UHPLC-QTOF-MS) method was developed to quantify major urinary metabolites simultaneously after testosterone intake. The sample preparation of the urine (1 mL) was performed by solid-phase extraction on Oasis HLB sorbent using a 96-well plate format. The conjugated steroids were analyzed by UHPLC-QTOF-MS(E) with a single-gradient elution of 36 min (including re-equilibration time) in the negative electrospray ionization mode. MS(E) analysis involved parallel alternating acquisitions of both low- and high-collision energy functions. The method was validated and applied to samples collected from a clinical study performed with a group of healthy human volunteers who had taken testosterone, which were compared with samples from a placebo group. Quantitative results were also compared to GC-MS and LC-MS/MS measurements, and the correlations between data were found appropriate. The acquisition of full mass spectra over the entire mass range with QTOF mass analyzers gives promise of the opportunity to extend the steroid profile to a higher number of conjugated steroids.

In vitro biotransformation of imatinib by the tumor expressed CYP1A1 and CYP1B1.

The main objective of the study was to examine the biotransformation of the anticancer drug imatinib in target cells by incubating it with oxidoreductases expressed in tumor cells. The second objective was to obtain an in silico prediction of the potential activity of imatinib metabolites. An in vitro enzyme kinetic study was performed with cDNA expressed human oxidoreductases and LC-MS/MS analysis. The kinetic parameters (Km and Vmax) were determined for six metabolites. A molecular modeling approach was used to dock these metabolites to the target Abl or Bcr-Abl kinases. CYP3A4 isozyme showed the broadest metabolic capacity, whereas CYP1A1, CYP1B1 and FMO3 isozymes biotransformed imatinib with a high intrinsic clearance. The predicted binding modes for the metabolites to Abl were comparable to that of the parent drug, suggesting potential activity. These findings indicate that CYP1A1 and CYP1B1, which are known to be overexpressed in a wide range of tumors, are involved in the biotransformation of imatinib. They could play a role in imatinib disposition in the targeted stem, progenitor and differentiated cancer cells, with a possible contribution of the metabolites toward the activity of the drug.

Alterações de atributos químicos e biológicos de solo e rendimento de milho e soja pela utilização de resíduos de curtume e carbonífero

Resíduos gerados por curtumes e pela exploração de carvão mineral são produtos potencialmente poluentes em várias regiões do Brasil, principalmente nos estados do Rio Grande do Sul e de Santa Catarina. O presente trabalho foi realizado no campo com o objetivo de avaliar o efeito da adição de resíduos de curtume e rejeito carbonífero sobre o solo e sobre as plantas cultivadas em um Argissolo Vermelho distrófico típico da Estação Experimental Agronômica da UFRGS, localizada no município de Eldorado do Sul (RS). O experimento foi realizado no ano agrícola de 1996/97, em parcelas de 70 m², cultivando-se soja (Glycine max L. Merrill) e milho (Zea mays L.). Tratamentos com a adição de 21,3 e 42,5 t ha-1 de lodo de curtume (LC) com adubação fosfatada e potássica na forma mineral foram comparados com o tratamento com adubação mineral completa (NPK) mais correção da acidez do solo e com a testemunha absoluta. Foram estudados também: (a) adição de resíduo carbonífero (106 t ha-1) mais adubação mineral; (b) resíduo carbonífero (106 t ha-1) mais lodo de curtume (21,3 t ha-1) com adubação fosfatada e potássica; (c) serragem cromada (29 t ha-1) mais adubação mineral (NPK), e (d) Cr mineral (125 kg ha-1) mais lodo de curtume (21,3 t ha-1) com adubação fosfatada e potássica. Foram avaliadas as alterações químicas e biológicas do solo, bem como os efeitos da aplicação dos resíduos sobre o rendimento e absorção de metais pelas plantas. A adição de LC aumentou o valor de pH e o teor de Ca trocável do solo. Não foram constatadas alterações nas concentrações dos metais Cu, Cd, Pb e Ni no solo, aumentando, entretanto, significativamente os teores de Cr. A atividade microbiana avaliada pela produção de CO2 foi estimulada pela adição dos resíduos, mas a população de bactérias, fungos e ctinomicetos não foi afetada. A adição de LC propiciou rendimentos de soja e milho semelhantes aos com adição de fertilizante nitrogenado. A adição de serragem cromada não alterou o rendimento das culturas, enquanto o rendimento de grãos de milho aumentou com a adição de resíduo carbonífero.

Eficiência de absorção, translocação e uso de cálcio, magnésio e enxofre por mudas enxertadas de Coffea arabica

O estudo da eficiência nutricional de plantas enxertadas de cafeeiro é importante para a seleção de combinações enxerto/porta-enxerto, visando ao desenvolvimento e produção máximos. O presente trabalho teve como objetivo avaliar, em cultivo hidropônico, a eficiência de absorção, translocação e uso de Ca, Mg e S por mudas enxertadas de Coffea arabica L., influenciada pelo porta-enxerto. O experimento foi realizado em casa de vegetação por um período de 170 dias, em vasos que continham areia, como substrato, e solução nutritiva circulante. Utilizaram-se, como enxerto, quatro genótipos de Coffea arabica L.: as variedades Catuaí Vermelho IAC 15 e Oeiras MG 6851 e as linhagens H 419-10-3-1-5 e H 514-5-5-3, e, como porta-enxerto, três genótipos de C. canephora Pierre et Froenher: Apoatã LC 2258, Conillon Muriaé-1, Robustão Capixaba (EMCAPA 8141) e um genótipo de Coffea arabica L.: Mundo Novo IAC 376-4, além da utilização de quatro pés-francos. Utilizou-se o delineamento experimental de blocos casualizados com quatro repetições. Os contrastes entre médias compararam as mudas de pé-franco com as associações enxerto/porta-enxerto. A eficiência de absorção, translocação e uso de Ca, Mg e S por mudas enxertadas de cafeeiro variou de acordo com a combinação enxerto/porta-enxerto. Somente a eficiência de translocação de Ca não foi alterada pela combinação enxerto/porta-enxerto. A linhagem H 514-5-5-3 foi beneficiada na eficiência de uso de Mg e produção de matéria seca pelos porta-enxertos Mundo Novo IAC 376-4 e Apoatã LC 2258, e na eficiência de uso de Ca e S apenas pelo Mundo Novo IAC 376-4.

Comparative protein profiling identifies elongation factor-1beta and tryparedoxin peroxidase as factors associated with metastasis in Leishmania guyanensis.

Parasites of the Leishmania Viannia subgenus are major causative agents of mucocutaneous leishmaniasis (MCL), a disease characterised by parasite dissemination (metastasis) from the original cutaneous lesion to form debilitating secondary lesions in the nasopharyngeal mucosa. We employed a protein profiling approach to identify potential metastasis factors in laboratory clones of L. (V.) guyanensis with stable phenotypes ranging from highly metastatic (M+) through infrequently metastatic (M+/M-) to non-metastatic (M-). Comparison of the soluble proteomes of promastigotes by two-dimensional electrophoresis revealed two abundant protein spots specifically associated with M+ and M+/M- clones (Met2 and Met3) and two others exclusively expressed in M- parasites (Met1 and Met4). The association between clinical disease phenotype and differential expression of Met1-Met4 was less clear in L. Viannia strains from mucosal (M+) or cutaneous (M-) lesions of patients. Identification of Met1-Met4 by biological mass spectrometry (LC-ES-MS/MS) and bioinformatics revealed that M+ and M- clones express distinct acidic and neutral isoforms of both elongation factor-1 subunit beta (EF-1beta) and cytosolic tryparedoxin peroxidase (TXNPx). This interchange of isoforms may relate to the mechanisms by which the activities of EF-1beta and TXNPx are modulated, and/or differential post-translational modification of the gene product(s). The multiple metabolic functions of EF-1 and TXNPx support the plausibility of their participation in parasite survival and persistence and thereby, metastatic disease. Both polypeptides are active in resistance to chemical and oxidant stress, providing a basis for further elucidation of the importance of antioxidant defence in the pathogenesis underlying MCL.

An improved trap design for decoupling multinuclear RF coils.

PURPOSE: Multinuclear magnetic resonance spectroscopy and imaging require a radiofrequency probe capable of transmitting and receiving at the proton and non-proton frequencies. To minimize coupling between probe elements tuned to different frequencies, LC (inductor-capacitor) traps blocking current at the (1) H frequency can be inserted in non-proton elements. This work compares LC traps with LCC traps, a modified design incorporating an additional capacitor, enabling control of the trap reactance at the low frequency while maintaining (1) H blocking. METHODS: Losses introduced by both types of trap were analysed using circuit models. Radiofrequency coils incorporating a series of LC and LCC traps were then built and evaluated at the bench. LCC trap performance was then confirmed using (1) H and (13) C measurements in a 7T human scanner. RESULTS: LC and LCC traps both effectively block interaction between non-proton and proton coils at the proton frequency. LCC traps were found to introduce a sensitivity reduction of 5±2%, which was less than half of that caused by LC traps. CONCLUSION: Sensitivity of non-proton coils is critical. The improved trap design, incorporating one extra capacitor, significantly reduces losses introduced by the trap in the non-proton coil. Magn Reson Med 72:584-590, 2014. © 2013 Wiley Periodicals, Inc.

Absorção, translocação e utilização de zinco, cobre e manganês por mudas enxertadas de Coffea arabica

Comparativamente ao pé-franco, a combinação enxerto/porta-enxerto altera os complexos mecanismos de "feedback" entre parte aérea e raízes, afetando de maneira positiva ou negativa a eficiência nutricional da planta. Este trabalho teve como objetivo avaliar, em cultivo hidropônico, a eficiência da absorção, translocação e utilização de Zn, Cu e Mn por mudas de Coffea arabica L., de acordo com o porta-enxerto utilizado. O experimento foi realizado em casa de vegetação, por um período de 170 dias, em vasos que continham areia como substrato, recebendo solução nutritiva circulante. Utilizaram-se, como enxerto, quatro genótipos de C. arabica: os cultivares Catuaí Vermelho IAC 15 e Oeiras MG 6851 e os híbridos 'H 419-10-3-1-5' e 'H 514-5-5-3' , e, como porta-enxerto, quatro genótipos, sendo três de Coffea canephora Pierre ex Froenher: Apoatã LC 2258, Conilon Muriaé-1 e RC EMCAPA 8141 (recombinação entre clones da variedade Robustão Capixaba - EMCAPA 8141) e uma linhagem de Coffea arabica L.: Mundo Novo IAC 376-4, além de quatro pés-francos. O delineamento experimental utilizado foi em blocos casualizados com 20 tratamentos, quatro repetições e uma planta por parcela. A eficiência nutricional das mudas quanto ao Zn, Cu e Mn variou de acordo com a combinação enxerto/porta-enxerto. A progênie 'H 514-5-5-3' foi mais eficiente quanto à utilização de Zn, Cu e Mn e produção de matéria seca, quando combinada com os porta-enxertos Apoatã LC 2258 e Mundo Novo IAC 376-4. O Catuaí Vermelho IAC 15 foi mais eficiente na utilização de Cu e Mn quando combinado com Apoatã LC 2258.

PROTEOMICS OF OXIDATIVE LESIONS OCCURRING DURING TRANSFUSION-PURPOSED STORAGE OF ERYTHROCYTE CONCENTRATES

Erythrocyte concentrates (ECs) are the major labile blood product being transfused worldwide, aiming at curing anemia of diverse origins. In Switzerland, ECs are stored at 4 °C up to 42 days in saline-adenine-glucose-mannitol (SAGM). Such storage induces cellular lesions, altering red blood cells (RBCs) metabolism, protein content and rheological properties. A hot debate exists regarding the impact of the storage lesions, thus the age of ECs on transfusion-related clinical adverse outcomes. Several studies tend to show that poorer outcomes occur in patients receiving older blood products. However, no clear association was demonstrated up to date. While metabolism and early rheological changes are reversible through transfusion of the blood units, oxidized proteins cannot be repaired, and it is likely such irreversible damages would affect the quality of the blood product and the efficiency of the transfusion. In vivo, RBCs are constantly exposed to oxygen fluxes, and are thus well equipped to deal with oxidative challenges. Moreover, functional 20S proteasome complexes allow for recognition and proteolysis of fairly oxidized protein, and some proteins can be eliminated from RBCs by the release of microvesicles. The present PhD thesis is involved in a global research project which goal is to characterize the effect of processing and storage on the quality of ECs. Assessing protein oxidative damages during RBC storage is of major importance to understand the mechanisms of aging of stored RBCs. To this purpose, redox proteomic-based investigations were conducted here. In a first part, cysteine oxidation and protein carbonylation were addressed via 2D-DIGE and derivatization-driven immunodetection approaches, respectively. Then, the oxidized sub- proteomes were characterized through LC-MS/MS identification of proteins in spots of interest (cysteine oxidation) or affinity-purified carbonylated proteins. Gene ontology annotation allowed classifying targets of oxidation according to their molecular functions. In a third part, the P20S activity was evaluated throughout the storage period of ECs, and its susceptibility to highly oxidized environment was investigated. The potential defensive role of microvesiculation was also addressed through the quantification of eliminated carbonylated proteins. We highlighted distinct protein groups differentially affected by cysteine oxidation, either reversibly or irreversibly. In addition, soluble extracts showed a decrease in carbonylation at the beginning of the storage and membrane extracts revealed increasing carbonylation after 4 weeks of storage. Engaged molecular functions revealed that antioxidant (AO) are rather reversibly oxidized at their cysteine residue(s), but are irreversibly oxidized through carbonylation. In the meantime, the 20S proteasome activity is decreased by around 40 % at the end of the storage period. Incubation of fresh RBCs extracts with exogenous oxidized proteins showed a dose-dependent and protein-dependent inhibitory effect. Finally, we proved that the release of microvesicles allows the elimination of increasing quantities of carbonylated proteins. Taken together, these results revealed an oxidative pathway model of RBCs storage, on which further investigation towards improved storage conditions will be based. -- Les concentrés érythrocytaires (CE) sont le produit sanguin le plus délivré au monde, permettant de traiter différentes formes d'anémies. En Suisse, les CE sont stocké à 4 °C pendant 42 jours dans une solution saline d'adénine, glucose et mannitol (SAGM). Une telle conservation induit des lésions de stockage qui altèrent le métabolisme, les protéines et les propriétés rhéologique du globule rouge (GR). Un débat important concerne l'impact du temps de stockage des CE sur les risques de réaction transfusionnelles, certaines études tentant de démontrer que des transfusions de sang vieux réduiraient l'espérance de vie des patients. Cependant, aucune association concrète n'a été prouvée à ce jour. Alors que les modifications du métabolisme et changement précoces des propriétés rhéologiques sont réversibles suite à la transfusion du CE, les protéines oxydées ne peuvent être réparées, et il est probable que de telles lésions affectent la qualité et l'efficacité des produits sanguins. In vivo, les GR sont constamment exposés à l'oxygène, et sont donc bien équipés pour résister aux lésions oxydatives. De plus, les complexes fonctionnels de proteasome 20S reconnaissent et dégradent les protéines modérément oxydées, et certaines protéines peuvent être éliminées par les microparticules. Cette thèse de doctorat est imbriquée dans un projet de recherche global ayant pour objectif la caractérisation des effets de la préparation et du stockage sur la qualité des GR. Evaluer les dommages oxydatifs du GR pendant le stockage est primordial pour comprendre les mécanismes de vieillissement des produits sanguin. Dans ce but, des recherches orientées redoxomique ont été conduites. Dans une première partie, l'oxydation des cystéines et la carbonylation des protéines sont évaluées par électrophorèse bidimensionnelle différentielle et par immunodétection de protéines dérivatisées. Ensuite, les protéines d'intérêt ainsi que les protéines carbonylées, purifiées par affinité, sont identifiées par spectrométrie de masse en tandem. Les protéines cibles de l'oxydation sont classées selon leur fonction moléculaire. Dans une troisième partie, l'activité protéolytique du protéasome 20S est suivie durant la période de stockage. L'impact du stress oxydant sur cette activité a été évalué en utilisant des protéines exogènes oxydées in vitro. Le potentiel rôle défensif de la microvesiculation a également été étudié par la quantification des protéines carbonylées éliminées. Dans ce travail, nous avons observé que différents groupes de protéines sont affectés par l'oxydation réversible ou irréversible de leurs cystéines. De plus, une diminution de la carbonylation en début de stockage dans les extraits solubles et une augmentation de la carbonylation après 4 semaines dans les extraits membranaires ont été montrées. Les fonctions moléculaires engagées par les protéines altérées montrent que les défenses antioxydantes sont oxydées de façon réversible sur leurs résidus cystéines, mais sont également irréversiblement carbonylées. Pendant ce temps, l'activité protéolytique du protéasome 20S décroit de 40 % en fin de stockage. L'incubation d'extraits de GR en début de stockage avec des protéines oxydées exogènes montre un effet inhibiteur « dose-dépendant » et « protéine-dépendant ». Enfin, les microvésicules s'avèrent éliminer des quantités croissantes de protéines carbonylées. La synthèse de ces résultats permet de modéliser une voie oxydative du stockage des GRs, à partir de laquelle de futures recherches seront menées avec pour but l'amélioration des conditions de stockage.