Redox remodelling in diaphragm muscle adaptation to chronic sustained hypoxia

Chronic sustained hypoxia (CH) induces functional weakness, atrophy, and mitochondrial remodelling in the diaphragm muscle. Animal models of CH present with changes similar to patients with respiratory-related disease, thus, elucidating the molecular mechanisms driving these adaptations is clinically important. We hypothesize that ROS are pivotal in diaphragm muscle adaptation to CH. C57BL6/J mice were exposed to CH (FiO2=0.1) for one, three, and six weeks. Sternohyoid (upper airway dilator), extensor digitorum longus (EDL), and soleus were studied as reference muscles as well as the diaphragm. The diaphragm was profiled using a redox proteomics approach followed by mass spectrometry. Following this, redox-modified metabolic enzyme activities and atrophy signalling were assessed using spectrophotometric assays and ELISA. Diaphragm isotonic performance was assessed after six weeks of CH ± chronic antioxidant supplementation. Protein carbonyl and free thiol content in the diaphragm were increased and decreased respectively after six weeks of CH – indicative of protein oxidation. These changes were temporally modulated and muscle specific. Extensive remodelling of metabolic proteins occurred and the stress reached the cross-bridge. Metabolic enzyme activities in the diaphragm were, for the most part, decreased by CH and differential muscle responses were observed. Redox sensitive chymotrypsin-like proteasome activity of the diaphragm was increased and atrophy signalling was observed through decreased phospho-FOXO3a and phospho-mTOR. Phospho-p38 MAPK content was increased and this was attenuated by antioxidant treatment. Hypoxia decreased power generating capacity of the diaphragm and this was restored by N-acetyl-cysteine (NAC) but not by tempol. Redox remodelling is pivotal for diaphragm adaptation to chronic sustained hypoxia. Muscle changes are dependent on duration of the hypoxia stimulus, activity profile of the muscle, and molecular composition of the muscle. The working respiratory muscles and slow oxidative fibres are particularly susceptible. NAC (antioxidant) may be useful as an adjunct therapy in respiratory-related diseases characterised by hypoxic stress.

Impacts of Mountaintop Removal Coal Mining on the Mud River, West Virginia: Selenium Accumulation, Trophic Transfer, and Toxicity in Fish

Selenium (Se) is a micronutrient necessary for the function of a variety of important enzymes; Se also exhibits a narrow range in concentrations between essentiality and toxicity. Oviparous vertebrates such as birds and fish are especially sensitive to Se toxicity, which causes reproductive impairment and defects in embryo development. Selenium occurs naturally in the Earth's crust, but it can be mobilized by a variety of anthropogenic activities, including agricultural practices, coal burning, and mining.

Mountaintop removal/valley fill (MTR/VF) coal mining is a form of surface mining found throughout central Appalachia in the United States that involves blasting off the tops of mountains to access underlying coal seams. Spoil rock from the mountain is placed into adjacent valleys, forming valley fills, which bury stream headwaters and negatively impact surface water quality. This research focused on the biological impacts of Se leached from MTR/VF coal mining operations located around the Mud River, West Virginia.

In order to assess the status of Se in a lotic (flowing) system such as the Mud River, surface water, insects, and fish samples including creek chub (Semotilus atromaculatus) and green sunfish (Lepomis cyanellus) were collected from a mining impacted site as well as from a reference site not impacted by mining. Analysis of samples from the mined site showed increased conductivity and Se in the surface waters compared to the reference site in addition to increased concentrations of Se in insects and fish. Histological analysis of mined site fish gills showed a lack of normal parasites, suggesting parasite populations may be disrupted due to poor water quality. X-ray absorption near edge spectroscopy techniques were used to determine the speciation of Se in insect and creek chub samples. Insects contained approximately 40-50% inorganic Se (selenate and selenite) and 50-60% organic Se (Se-methionine and Se-cystine) while fish tissues contained lower proportions of inorganic Se than insects, instead having higher proportions of organic Se in the forms of methyl-Se-cysteine, Se-cystine, and Se-methionine.

Otoliths, calcified inner ear structures, were also collected from Mud River creek chubs and green sunfish and analyzed for Se content using laser ablation inductively couple mass spectrometry (LA-ICP-MS). Significant differences were found between the two species of fish, based on the concentrations of otolith Se. Green sunfish otoliths from all sites contained background or low concentrations of otolith Se (< 1 µg/g) that were not significantly different between mined and unmined sites. In contrast creek chub otoliths from the historically mined site contained much higher (≥ 5 µg/g, up to approximately 68 µg/g) concentrations of Se than for the same species in the unmined site or for the green sunfish. Otolith Se concentrations were related to muscle Se concentrations for creek chubs (R2 = 0.54, p = 0.0002 for the last 20% of the otolith Se versus muscle Se) while no relationship was observed for green sunfish.

Additional experiments using biofilms grown in the Mud River showed increased Se in mined site biofilms compared to the reference site. When we fed fathead minnows (Pimephales promelas) on these biofilms in the laboratory they accumulated higher concentrations of Se in liver and ovary tissues compared to fathead minnows fed on reference site biofilms. No differences in Se accumulation were found in muscle from either treatment group. Biofilms were also centrifuged and separated into filamentous green algae and the remaining diatom fraction. The majority of Se was found in the diatom fraction with only about 1/3rd of total biofilm Se concentration present in the filamentous green algae fraction

Finally, zebrafish (Danio rerio) embryos were exposed to aqueous Se in the form of selenate, selenite, and L-selenomethionine in an attempt to determine if oxidative stress plays a role in selenium embryo toxicity. Selenate and selenite exposure did not induce embryo deformities (lordosis and craniofacial malformation). L-selenomethionine, however, induced significantly higher deformity rates at 100 µg/L compared to controls. Antioxidant rescue of L-selenomethionime induced deformities was attempted in embryos using N-acetylcysteine (NAC). Pretreatment with NAC significantly reduced deformities in the zebrafish embryos secondarily treated with L-selenomethionine, suggesting that oxidative stress may play a role in Se toxicity. Selenite exposure also induced a 6.6-fold increase in glutathione-S-transferase pi class 2 gene expression, which is involved in xenobiotic transformation. No changes in gene expression were observed for selenate or L-selenomethionine-exposed embryos.

The findings in this dissertation contribute to the understanding of how Se bioaccumulates in a lotic system and is transferred through a simulated foodweb in addition to further exploring oxidative stress as a potential mechanism for Se-induced embryo toxicity. Future studies should continue to pursue the role of oxidative stress and other mechanisms in Se toxicity and the biotransformation of Se in aquatic ecosystems.

Metabolic programming and PDHK1 control CD4+ T cell subsets and inflammation.

Activation of CD4+ T cells results in rapid proliferation and differentiation into effector and regulatory subsets. CD4+ effector T cell (Teff) (Th1 and Th17) and Treg subsets are metabolically distinct, yet the specific metabolic differences that modify T cell populations are uncertain. Here, we evaluated CD4+ T cell populations in murine models and determined that inflammatory Teffs maintain high expression of glycolytic genes and rely on high glycolytic rates, while Tregs are oxidative and require mitochondrial electron transport to proliferate, differentiate, and survive. Metabolic profiling revealed that pyruvate dehydrogenase (PDH) is a key bifurcation point between T cell glycolytic and oxidative metabolism. PDH function is inhibited by PDH kinases (PDHKs). PDHK1 was expressed in Th17 cells, but not Th1 cells, and at low levels in Tregs, and inhibition or knockdown of PDHK1 selectively suppressed Th17 cells and increased Tregs. This alteration in the CD4+ T cell populations was mediated in part through ROS, as N-acetyl cysteine (NAC) treatment restored Th17 cell generation. Moreover, inhibition of PDHK1 modulated immunity and protected animals against experimental autoimmune encephalomyelitis, decreasing Th17 cells and increasing Tregs. Together, these data show that CD4+ subsets utilize and require distinct metabolic programs that can be targeted to control specific T cell populations in autoimmune and inflammatory diseases.

Participación de la familia génica GPC sobre la concentración de proteínas y nutrientes en el grano y su relación con la senescencia foliar en plantas de trigo

La removilización de nutrientes al grano y la senescencia son procesos extremadamente interconectados en trigo que determinan la calidad nutricional y panadera. Mientras que el nitrógeno (N) afecta la concentración de proteínas en grano (CPG), en donde alta CPG generan productos panificables con una calidad superior, los micronutrientes como el hierro (Fe) y el Zinc (Zn) afectan la calidad nutricional. Por lo tanto, el mejoramiento de la calidad en trigo requiere un profundo entendimiento de las redes génicas que regulan y controlan la senescencia y la removilización de nutrientes al grano. El gen GPC-B1, proveniente del trigo silvestre Triticum turgidum var. diccocoides, pertenece a la familia de factores de transcripción NAC y es la primer fuente de variación para CPG y micronutrientes identificada en trigo. Los objetivos de esta tesis fueron: estudiar el efecto de la introgresión de GPC-B1 sobre la CPG, concentración de micro y macronutrientes y diferentes parámetros agronómicos en germoplasma argentino; profundizar, mediante análisis transcriptómicos y el uso de plantas mutantes para los genes GPC el entendimiento de la regulación génica que ocurre durante la senescencia y finalmente identificar nuevos genes de transporte de N, Fe y Zn al grano mediante genómica comparativa con arroz. Cuando GPC-B1 se introdujo en germoplasma de origen argentino la CPG se incrementó entre 3 a 8,11 g kg-1 a través de diferentes cultivares y ambientes. A pesar del efecto negativo del gen sobre el tamaño de granos y una aceleración en la senescencia, no se observaron diferencias significativas en rendimiento. El incremento en la CPG se explicó por una mayor proteólisis y eficiencia en el transporte de aminoácidos al grano. Cuando se analizó la concentración de nutrientes, se observaron incrementos consistentes en Fe. La combinación de análisis transcriptómicos y plantas mutantes permitió identificar 3888 genes diferencialmente expresados durante la senescencia y 340 genes modulados por la familia GPC. A su vez, se han identificado 21 genes relacionados con el transporte de N al grano y se han caracterizado nueve familias génicas relacionadas con el transporte de Fe y Zn al grano que pueden utilizarse en programas de mejoramiento para incrementar la calidad nutricional en trigo.

North Atlantic and North Sea climate change: curl up, shut down, NAO and ocean colour

The strength of the North Atlantic Current (NAC) (based on sea-surface elevation sloped derived from altimeter data) is correlated with westerly winds (based on North Atlantic Oscillation [NAO] Index data over a nine year period [1992-2002] with 108 monthly values). The data time window includes the major change in climate forcing over the last 100 years (1995 to 1996). It is shown that the NAO Index can be used for early earning of system failure for the NAC. The correlation response or early warning time scale for western Europe and south England is six months. The decay scale for the NAC and Subtropical Gyre circulation is estimated as three years. Longer period altimeter elevation/circulation changes are discussed. The sea-surface temperature (SST) response of the North Sea to negative and positive NAO conditions is examined. The overall temperature response for the central North Sea to NAO index forcing, reflecting wind induced inflow, shelf circulation and local climate forcing, is similar to 5 months. In years with strong North Atlantic winter wind induced inflow, under marked NAO positive conditions, mean temperatures ( similar to 10.5 degree C) are about 1 degree C warmer than under negative conditions. In 1996 under extreme negative winter NAO conditions, the North Sea circulation stopped, conditions near the Dogger Bank became more continentally influenced and the winter (March) temperature fell to 3.1 degree C whereas in 1995 under NAO positive winter conditions the minimum temperature was 6.4 degree C (February). Seasonal advance of North Atlantic and North Sea temperature is derived in relation to temperature change. Temperature change and monthly NAO Index are discussed with respect to phytoplankton blooms, chlorophyll-a measurements, ocean colour data and the anomalous north-eastern Atlantic 2002 spring/summer bloom SeaWiFS chlorophyll concentrations.

Thermal front variability along the North Atlantic Current observed using satellite microwave and infrared data

Thermal fronts detected using multiple satellite sensors have been integrated to provide new information on the spatial and seasonal distribution of oceanic fronts in the North Atlantic. The branching of the North Atlantic Current (NAC) as it encounters the Mid-Atlantic Ridge (MAR) is reflected in surface thermal fronts, which preferentially occur at the Charlie Gibbs Fracture Zone (CGFZ) and several smaller fracture zones. North of the CGFZ there are few thermal fronts, contrasting with the region to the south, where there are frequent surface thermal fronts that are persistent seasonally and interannually. The alignment of the fronts confirms that the shallower Reykjanes Ridge north of the CGFZ is more of a barrier to water movements than the ridge to the south. Comparison of front distributions with satellite altimetry data indicates that the MAR influence on deep ocean currents is also frequently exhibited in surface temperature. The improved spatial and temporal resolution of the front analysis has revealed consistent seasonality in the branching patterns. These results contribute to our understanding of the variability of the NAC, and the techniques for visualising oceanic fronts can be applied in other regions to reveal details of surface currents that cannot be resolved using satellite altimetry or in situ measurements.

Seasonality and interannual variability of the European Slope Current from 20 years of altimeter data compared with in situ measurements

The European Slope Current (SC) is a major section of the warm poleward flow from the Atlantic to the Arctic, which also moderates the exchange of heat, salt, nutrients and carbon between the deep ocean and the European shelf seas. The mean structure of the geostrophic flow, seasonality, interannual variability and long-term trend of SC are appraised with an unprecedented continuous 20-year satellite altimeter dataset. Comparisons with long term in situ data showed a maximum correlation of r2=0.51 between altimeter and Acoustic Doppler Current Profilers (ADCP), with similar results for drogued buoy data. Mean geostrophic currents were appraised more comprehensively than previous attempts, and the paths of 4 branches of the North Atlantic Current (NAC) and positions of 5 eddies in the region were derived quantitatively. A consistent seasonal cycle in the flow of the SC was found at all 8 sections along the European shelf slope, with maximum poleward flow in the winter and minimum in the summer. The seasonal difference in the altimetry current speed amounted to ~8-10 cm s-1 at the northern sections, but only ~5 cm s-1 on the Bay of Biscay slopes. This extended altimeter dataset indicates significant regional and seasonal variations, and has revealed new insights into the interannual variability of the SC. It is shown that there is a peak poleward flow at most positions along a ~2000 km stretch of the continental slope from Portugal to Scotland during 1995-1997, but this did not clearly relate to the extreme negative North Atlantic Oscillation (NAO) in the winter of 1995-1996. The speed of the SC exhibited a long term decreasing trend of ~1% per year. By contrast the NAC showed no significant trend over the 20-year period. Major changes in the NAC occurred three times, and these changes followed decreases in the NAO index.

Nomenclature and syntaxonomic notes on some high-rank syntaxa of the European grassland vegetation

We present descriptions of a new order (Ranunculo cortusifolii-Geranietalia reuteri and of a new alliance (Stachyo lusitanicae-Cheirolophion sempervirentis) for the herbaceous fringe communities of Macaronesia and of the southwestern Iberian Peninsula, respectively. A new alliance, the Polygalo mediterraneae-Bromion erecti (mesophilous post-cultural grasslands), was introduced for the Peninsular Italy. We further validate and typify the Armerietalia rumelicae (perennial grasslands supported by nutrient-poor on siliceous bedrocks at altitudes characterized by the submediterranean climate of central-southern Balkan Peninsula), the Securigero-Dasypyrion villosae (lawn and fallow-land tall-grass annual vegetation of Italy), and the Cirsio vallis-demoni-Nardion (acidophilous grasslands on siliceous substrates of the Southern Italy). Nomenclatural issues (validity, legitimacy, synonymy, formal corrections) have been discussed and clarified for the following names: Brachypodio-Brometalia, Bromo pannonici-Festucion csikhegyensis, Corynephoro-Plantaginion radicatae, Heleochloion, Hieracio-Plantaginion radicatae, Nardetea strictae, Nardetalia strictae, Nardo-Callunetea, Nardo-Galion saxatilis, Oligo-Bromion, Paspalo-Heleochloetalia, Plantagini-Corynephorion and Scorzoneret alia villosae. 

alpha-Synuclein aggregation

Alpha-synuclein is a major component of Lewy bodies in Parkinson's disease and is found associated with several other forms of dementia. As with other neurodegenerative diseases, the ability of alpha-synuclein to aggregate and form fibrillar deposits seems central to its pathology. We have defined a sequence within the NAC region of alpha-synuclein that is necessary for aggregation. Exploitation of chemically modified analogues of this peptide may produce inhibitors of aggregation.

Aggregation and neurotoxicity of alpha-synuclein and related peptides

Fibrillar deposits of alpha-synuclein occur in several neurodegenerative diseases. Two mutant forms of alpha-synuclein have been associated with early-onset Parkinson's disease, and a fragment has been identified as the non-amyloid-beta peptide component of Alzheimer's disease amyloid (NAC). Upon aging, solutions of alpha-synuclein and NAC change conformation to beta-sheet, detectable by CD spectroscopy, and form oligomers that deposit as amyloid-like fibrils, detectable by electron microscopy. These aged peptides are also neurotoxic. Experiments on fragments of NAC have enabled the region of NAC responsible for its aggregation and toxicity to be identified. NAC(8-18) is the smallest fragment that aggregates, as indicated by the concentration of peptide remaining in solution after 3 days, and forms fibrils, as determined by electron microscopy. Fragments NAC(8-18) and NAC(8-16) are toxic, whereas NAC(12-18), NAC(9-16) and NAC(8-15) are not. Hence residues 8-16 of NAC comprise the region crucial for toxicity. Toxicity induced by alpha-synuclein, NAC and NAC(1-18) oligomers occurs via an apoptotic mechanism, possibly initiated by oxidative damage, since these peptides liberate hydroxyl radicals in the presence of iron. Molecules with anti-aggregational and/or antioxidant properties may therefore be potential therapeutic agents.

Sensory neuropeptides induce histamine release from bronchoalveolar lavage cells in both nonasthmatic coughers and cough variant asthmatics

BACKGROUND: Sensory neuropeptides have been suggested to play a role in the pathogenesis of a number of respiratory diseases including asthma and chronic non-productive cough.

OBJECTIVES: To investigate the action of sensory neuropeptides on airway mast cells obtained by bronchoalveolar lavage (BAL).

METHODS: BAL was performed on 23 nonasthmatic patients with cough (NAC), 11 patients with cough variant asthma (CVA) and 10 nonatopic controls. Washed lavage cells were stimulated (20 min, 37 degrees C) with calcitonin gene-related peptide (CGRP), neurokinin A (NKA) and substance P (25 and 50 micromol/L).

RESULTS: The neuropeptides tested induced histamine release in all groups studied. Only CGRP (50 micromol/L) induced significantly more histamine release from both NAC and CVA patients compared with control subjects (P = 0.038 and 0.045, respectively).

CONCLUSION: Regardless of aetiology, mast cells from patients with chronic cough appear to have an increased responsiveness to CGRP compared with controls. The results of the present study suggest that the role of CGRP in chronic cough should be further investigated.

Optimizing The Nasal Allergen Challenge Model For The Study Of Allergic Rhinitis

Background The Allergic Rhinitis Clinical Investigator Collaborative (AR-CIC) uses a Nasal Allergen Challenge (NAC) model to study the pathophysiology of AR and provides proof of concept for novel therapeutics. The NAC model needs to ensure optimal participant qualification, allergen challenge, clinical symptoms capture and biological samples collection. Repeatability of the protocol is key to ensuring unbiased efficacy analysis of novel therapeutics. The effect of allergen challenge on IL-33 gene expression and its relation to IL1RL1 receptor and cytokine secretion was investigated. Methods Several iterations of the NAC protocol was tested, comparing variations of qualifying criteria based on the Total Nasal Symptom Score (TNSS) and Peak Nasal Inspiratory Flow (PNIF). The lowest allergen concentration was delivered and TNSS and PNIF recorded 15 minutes later. Participants qualified if the particular criteria for the protocol were met, otherwise the next higher allergen concentration (4-fold increase), was administered until the targets were reached. Participants returned for a NAC visit and received varying allergen challenge concentrations depending on the protocol, TNSS/PNIF were recorded at 15 minutes, 30 minutes, 1 hour, and hourly up to 12 hours, a 24 hour time point was added in later iterations. Repeatability was evaluated using a 3-4week interval between screening, NAC1, and NAC2 visits. Various biomarker samples were collected. Results A combined TNSS and PNIF criterion was more successful in qualifying participants. The cumulative allergen challenge (CAC) protocol proved more reliable in producing a robust clinical and biomarker response. Repeatability of the CAC protocol was achieved with a 3-week interval between visits, on a clinical and biological basis. IL-33 cytokine is an important biomarker in initiating the inflammatory response in AR in humans. IL-33 and IL1RL1 expression might employ a negative feedback mechanism in human nasal epithelial cells. Comparing the clinical and biological response to ragweed vs cat allergen challenge, proved the CAC protocol’s suitability for use employing different allergens. Conclusion The AR-CIC’s CAC protocol is an effective method of studying AR, capable of generating measurable and repeatable clinical and biomarker responses, enabling better understanding of AR pathophysiology and ensuring that any change would be purely due to medication under investigation in a clinical trial setting.

A test for mucus removal in the chiton Lepidochitona cinerea (Linnaeus, 1767) (Polyplacophora: Chitonida: Ischnochitonidae)

Several methods have been proposed to ‘clean’ the soft tissues of molluscs of mucus, so that the surface cilia can be examined microscopically. We report the first empirical test of the effectiveness of methods for removing mucus in the pallial cavity surface of chitons. Three methods were compared, at several time intervals: the enzyme hyaluronidase, the mucolytic agent N-acetyl cysteine (NAC), and seawater washing via the natural action of cilia in excised tissue. Treatment in NAC for 10 min produced the best results, and we recommend this protocol as a starting point for further investigation on mucus removal in a broader suite of taxa. We present the first description of the pallial surface cilia in the chiton Lepidochitona cinerea. During the course of this study, we also determined that these chitons were frequently infested with a ciliate protozoan parasite, Trichodina sp., which have been historically reported from chitons but never studied in detail. The parasites were absent where antimucus treatments were effective, but their abundance and large size (about 30-mm diameter) in less successful treatments obscured the view of the pallial cavity surface.

Assessment of a zoomed global model for the North Sea by comparison with a conventional nested regional model

Assessment of marine downscaling of global model simulations to the regional scale is a prerequisite for understanding ocean feedback to the atmosphere in regional climate downscaling. Major difficulties arise from the coarse grid resolution of global models, which cannot provide sufficiently accurate boundary values for the regional model. In this study, we first setup a stretched global model (MPIOM) to focus on the North Sea by shifting poles. Second, a regional model (HAMSOM) was performed with higher resolution, while the open boundary values were provided by the stretched global model. In general, the sea surface temperatures (SSTs) in the two experiments are similar. Major SST differences are found in coastal regions (root mean square difference of SST is reaching up to 2°C). The higher sea surface salinity in coastal regions in the global model indicates the general limitation of this global model and its configuration (surface layer thickness is 16 m). By comparison, the advantage of the absence of open lateral boundaries in the global model can be demonstrated, in particular for the transition region between the North Sea and Baltic Sea. On long timescales, the North Atlantic Current (NAC) inflow through the northern boundary correlates well between both model simulations (R~0.9). After downscaling with HAMSOM, the NAC inflow through the northern boundary decreases by ~10%, but the circulation in the Skagerrak is stronger in HAMSOM. The circulation patterns of both models are similar in the northern North Sea. The comparison suggests that the stretched global model system is a suitable tool for long-term free climate model simulations, and the only limitations occur in coastal regions. Regarding the regional studies focusing on the coastal zone, nested regional model can be a helpful alternative.

Vieillissement vasculaire chez des patients athérosclérotiques: Sénescence prématurée des cellules endothéliales?

La dysfonction de l’endothélium vasculaire, associée à une diminution de ses propriétés vasorelaxantes et anti-thrombogéniques, survient avec le vieillissement mais également chez de plus jeunes patients athérosclérotiques présentant plusieurs facteurs de risque cardiovasculaire. Au niveau cellulaire, le vieillissement des cellules endothéliales (CE) mène à un état irréversible de non division cellulaire appelé sénescence. Ces cellules sénescentes présentent des changements spécifiques au niveau de leur morphologie et de l’expression génique, menant à leur dysfonction. La sénescence dite réplicative est déclenchée par le raccourcissement des télomères survenant à chaque division cellulaire, mais peut également être induite prématurément par le stress oxydant (SIPS). L’objectif principal de cette étude est de caractériser la sénescence de CE vasculaires isolées à partir de patients athérosclérotiques, et d’observer l’impact des facteurs de risque sur cette sénescence. Afin de confirmer la contribution des deux principales voies de la sénescence, nous avons par la suite étudié conjointement ou séparément, l’impact d’un traitement chronique avec un antioxydant sur la sénescence de CE, et d’une surexpression de la sous-unité catalytique de la télomérase (hTERT), une enzyme responsable de l’allongement des télomères. Nous avons isolé et cultivé des CE provenant d’artères mammaires internes prélevées lors de pontages coronariens. Selon les études, les cellules ont été infectées ou non avec un lentivirus surexprimant la hTERT, et cultivées in vitro jusqu’à sénescence, en présence ou en absence de l’antioxydant N-acétyl-L-cystéine (NAC). Différents marqueurs des deux principales voies de la sénescence (réplicative ou SIPS) ont été quantifiés. La sénescence cellulaire se développe exponentiellement avec le temps et est associée à une réduction de la viabilité et de la prolifération cellulaires. Chez les patients athérosclérotiques, le vieillissement des CE passe par les deux principales voies de la sénescence : des télomères courts initialement en culture et la durée d’exposition in vivo aux facteurs de risque cardiovasculaire prédisent une apparition prématurée de la sénescence. Toutefois, chez les fumeurs, la sénescence est exclusivement du type SIPS. Ces facteurs de risque cardiovasculaire et principalement l’hypertension, semblent accélèrer le vieillissement biologique et favoriser la dysfonction des CE. Lorsque traitées chroniquement avec le NAC, les CE présentant initialement de moindres dommages cellulaires et moléculaires ainsi qu’une meilleure défense antioxydante développent une sénescence retardée. Lorsque le NAC est combiné à une surexpression de la hTERT, les deux voies de la sénescence sont bloquées et une immortalisation cellulaire est observée. À l’inverse, dans les CE les plus endommagées par les ROS in vivo, le NAC n’a aucun effet sur le développement de la sénescence, la hTERT, seule ou en combinaison avec le NAC, retarde légèrement la sénescence mais aucune immortalisation n’est observée lorsque ces traitements sont combinés. En conclusion, nos études démontrent que l’exposition chronique au stress oxydant associé aux facteurs de risque cardiovasculaire accélère le développement de la sénescence de CE vasculaires, contribuant potentiellement à l’athérogénèse. Dans les cellules de patients athérosclérotiques, il semble exister un seuil de dommages cellulaires et moléculaires subis in vivo au-delà duquel, aucun traitement (antioxydant ou hTERT) ne peut être bénéfique.