Добірка наукової літератури з теми "Clot structure"

Abstract Introduction: Inorganic polyphosphate (polyP) is a negatively charged polymer of phosphate units linked by high energy phosphoanhydride bonds. Dense granules of human platelets contain polyP which is released in response to thrombin stimulation. We recently reported that polyphosphate is a potent hemostatic regulator, accelerating blood clotting by activating the contact pathway and promoting the activation of factor V. Our previous studies found that polyP did not affect the time to clot formation when plasma was clotted with thrombin, however, suggesting that polyP exerts its procoagulant actions upstream of thrombin. We now report that polyP enhances fibrin clot structure. Methods: Purified fibrinogen and polyP were preincubated for 15 min in multiwell plates in buffer containing CaCl2, after which clotting was initiated by adding 0.1 to 8 nM thrombin and fibrin clot formation was evaluated by quantifying the change in turbidity (A405). Mass-length ratios were calculated from scans of A400 to A800. The effect of polyP on fibrinolysis was examined by adding 8 nM plasmin to the reaction mixtures immediately prior to thrombin. Scanning electron microscopy (SEM) was employed to visualize clot structure, and time courses of covalent fibrin cross-linking were assessed by SDS-PAGE. Results: PolyP had no effect on time to clot formation, but clots formed in the presence of polyP had markedly (up to threefold) higher turbidity than clots formed in the absence of polyP (see figure), irrespective of thrombin concentration. The increased turbidity in the presence of polyP was calcium-dependent and was enhanced when fibrinogen, CaCl2, and polyP were preincubated for up to 15 min prior to initiation of clotting with thrombin. PolyP increased the mass-length ratio of fibrin, and SEM confirmed that fibers formed with polyP were thicker than those formed without polyP. The ability of polyP to enhance fibrin clot turbidity was independent of factor XIIIa activity, and polyP did not alter the rate or extent of covalent fibrin cross-linking by factor XIIIa. When plasmin was included in clotting reactions containing polyP, mean times to 50% clot lysis were 28.5 ± 0.8 min for clots without polyP but 120.4 ± 5.6 min for clots with polyP. Conclusions: PolyP alters polymerization of fibrin, resulting in fibers of higher mass-length ratio that are lysed more slowly. This effect is calcium-dependent and is enhanced by preincubation of fibrinogen with calcium and polyP. Release of polyP from activated platelets or infectious microorganisms may therefore enhance fibrin clot structure. Figure Figure

AbstractThe formation of a fibrin clot matrix plays a critical role in promoting hemostasis and wound healing. Fibrin dynamics can become disadvantageous in the formation of aberrant thrombus development. Structural characteristics of clots, such as fiber diameter, clot density, stiffness, or permeability, can determine overall clot integrity and functional characteristics that have implications on coagulation and fibrinolysis. This review examines known factors that contribute to changes in clot structure and the presence of structural clot changes in various disease states. These insights provide valuable information in forming therapeutic strategies for disease states where alterations in clot structure are observed. Additionally, the implications of structural changes in clot networks on bleeding and thrombus development in terms of disease states and clinical outcomes are also examined in this review.

AbstractIndividuals with elevated prothrombin levels are at increased risk of venous thrombosis. To understand the mechanism behind this observation, we studied the effect of prothrombin concentration on thrombin generation and fibrin clot structure. The pattern of thrombin generation was directly related to the prothrombin level at all concentrations tested. From 0% to 300% of normal plasma levels of prothrombin, increasing the prothrombin concentration increased the initial rate, peak, and total amount of thrombin generated. Importantly, fibrin clot structure was also affected by the prothrombin concentration. Fibrin clots made from prothrombin concentrations less than 10% of plasma levels were weak and poorly formed. Fibrin clots made at 10% to 100% of plasma levels of prothrombin had similar fiber structures (mass-to-length ratio; μ). However, the fiber mass-to-length ratio decreased with increasing prothrombin levels more than 100% of plasma levels, in a dose-dependent manner. These results suggest that increased levels of prothrombin alter thrombin generation and clot structure. Specifically, elevated prothrombin levels produce clots with reduced fibrin mass-to-length ratios compared with normal clots. We hypothesize that this alteration in fibrin clot structure is an important determinant of the risk of thrombosis.

Abstract Polyphosphate, a linear polymer of inorganic phosphate, is present in platelet dense granules and is secreted on platelet activation. We recently reported that polyphosphate is a potent hemostatic regulator, serving to activate the contact pathway of blood clotting and accelerate factor V activation. Because polyphosphate did not alter thrombin clotting times, it appeared to exert all its procoagulant actions upstream of thrombin. We now report that polyphosphate enhances fibrin clot structure in a calcium-dependent manner. Fibrin clots formed in the presence of polyphosphate had up to 3-fold higher turbidity, had higher mass-length ratios, and exhibited thicker fibers in scanning electron micrographs. The ability of polyphosphate to enhance fibrin clot turbidity was independent of factor XIIIa activity. When plasmin or a combination of plasminogen and tissue plasminogen activators were included in clotting reactions, fibrin clots formed in the presence of polyphosphate exhibited prolonged clot lysis times. Release of polyphosphate from activated platelets or infectious microorganisms may play an important role in modulating fibrin clot structure and increasing its resistance to fibrinolysis. Polyphosphate may also be useful in enhancing the structure of surgical fibrin sealants.

SummaryWe tested the hypothesis that plasma fibrin clot structure/function is unfavourably altered in patients with antiphospholipid syndrome (APS). Ex vivo plasma clot permeability, turbidity and susceptibility to lysis were determined in 126 consecutive patients with APS enrolled five months or more since thrombotic event vs 105 controls. Patients with both primary and secondary APS were characterised by 11% lower clot permeability (p<0.001), 4.8% shorter lag phase (p<0.001), 10% longer clot lysis time (p<0.001), and 4.7% higher maximum level of D-dimer released from clots (p=0.02) as compared to the controls. Scanning electron microscopy images confirmed denser fibrin networks composed of thinner fibres in APS. Clots from patients with “triple-antibody positivity” were formed after shorter lag phase (p=0.019) and were lysed at a slower rate (p=0.004) than in the remainder. Clots from APS patients who experienced stroke and/or myocardial infarction were 8% less permeable (p=0.01) and susceptible to lysis (10.4% longer clot lysis time [p=0.006] and 4.5% slower release of D-dimer from clots [p=0.01]) compared with those following venous thromboembolism alone. Multivariate analysis adjusted for potential confounders showed that in APS patients, lupus anticoagulant and “triple-positivity” were the independent predictors of clot permeability, while “triple-positivity” predicted lysis time. We conclude that APS is associated with prothrombotic plasma fibrin clot phenotype, with more pronounced abnormalities in arterial thrombosis. Molecular background for this novel prothrombotic mechanism in APS remains to be established.

Clot retraction, measured by serum expression, is absent in some cases of multiple myeloma. Decreased clot retraction has been attributed to platelet dysfunction. A new instrument allows simultaneous measurement of platelet-mediated force development during clot retraction and of clot elastic modulus. We report 10 patients with immunoglobulin (Ig) G myeloma in whom the abnormalities of fibrin structure were quantitatively defined and platelet-fibrin interactions were assessed. Fiber mass-to-length ratios were calculated from gel turbidity. Platelet force development and clot elastic modula were measured in platelet-rich plasma gels. Fiber mass-to-length ratios for IgG myeloma patients were smaller (means +/- SE) (0.98 +/- 0.19 x 10(13) Da/cm) than for normal controls (1.36 +/- 0.06 x 10(13) Da/cm), indicating thinner fiber formation. Elastic modula of myeloma clots (51,013 +/- 14,660 dyn/cm2) were strikingly larger than modula for normal controls (23,355 +/- 1,887 dyn/cm2), indicating that such clots are mechanically less flexible. Platelet force development 1,200 s after thrombin addition was not diminished in myeloma patients (8,315 +/- 1,155 dyn) vs. controls (6,906 +/- 606 dyn). Abnormal clot retraction in myeloma appears to be primarily due to altered clot structure rather than platelet dysfunction.

SummaryAlthough many in vitro fibrin studies are performed with plasma, in vivo clots and thrombi contain erythrocytes, or red blood cells (RBCs).To determine the effects of RBCs on fibrin clot structure and mechanical properties, we compared plasma clots without RBCs to those prepared with low (2 vol%), intermediate (5-10 vol%), or high (≥20 vol%) numbers of RBCs. By confocal microscopy, we found that low RBC concentrations had little effect on clot structure. Intermediate RBC concentrations caused heterogeneity in the fiber network with pockets of densely packed fibers alongside regions with few fibers. With high levels of RBCs, fibers arranged more uniformly but loosely around the cells. Scanning electron micrographs demonstrated an uneven distribution of RBCs throughout the clot and a significant increase in fiber diameter upon RBC incorporation. While permeability was not affected by RBC addition, at 20% or higher RBCs, the ratio of viscous modulus (G′′) to elastic modulus (G′) increased significantly over that of a clot without any RBCs. RBCs triggered variability in the fibrin network structure, individual fiber characteristics, and overall clot viscoelasticity compared to the absence of cells. These results are important for understanding in vivo clots and thrombi.

SummaryZinc released from activated platelets binds fibrin(ogen) and attenuates fibrinolysis. Although zinc also affects clot formation, the mechanism and consequences are poorly understood. To address these gaps, the effect of zinc on clot formation and structure was examined in the absence or presence of factor (F) XIII. Zinc accelerated a) plasma clotting by 1.4-fold, b) fibrinogen clotting by 3.5- and 2.3-fold in the absence or presence of FXIII, respectively, c) fragment X clotting by 1.3-fold, and d) polymerisation of fibrin monomers generated with thrombin or batroxobin by 2.5– and 1.8-fold, respectively. Whereas absorbance increased up to 3.3-fold when fibrinogen was clotted in the presence of zinc, absorbance of fragment X clots was unaffected by zinc, consistent with reports that zinc binds to the αC-domain of fibrin(ogen). Scanning electron microscopic analysis revealed a twofold increase in fibre diameter in the presence of zinc and in permeability studies, zinc increased clot porosity by 30-fold with or without FXIII. Whereas FXIII increased clot stiffness from 128 ± 19 Pa to 415 ± 27 Pa in rheological analyses, zinc reduced clot stiffness by 10– and 8.5-fold in the absence and presence of FXIII, respectively. Clots formed in the presence of zinc were more stable and resisted rupture with or without FXIII. Therefore, zinc accelerates clotting and reduces fibrin clot stiffness in a FXIII-independent manner, suggesting that zinc may work in concert with FXIII to modulate clot strength and stability.

The factors that contribute to pulmonary embolism (PE), a potentially fatal complication of deep vein thrombosis (DVT), remain poorly understood. Whereas fibrin clot structure and functional properties have been implicated in the pathology of venous thromboembolism and the risk for cardiovascular complications, their significance in PE remains uncertain. Therefore, we systematically compared and quantified clot formation and lysis time, plasminogen levels, viscoelastic properties, activated factor XIII cross-linking, and fibrin clot structure in isolated DVT and PE subjects. Clots made from plasma of PE subjects showed faster clot lysis times with no differences in lag time, rate of clot formation, or maximum absorbance of turbidity compared with DVT. Differences in lysis times were not due to alterations in plasminogen levels. Compared with DVT, clots derived from PE subjects showed accelerated establishment of viscoelastic properties, documented by a decrease in lag time and an increase in the rate of viscoelastic property formation. The rate and extent of fibrin cross-linking by activated factor XIII were similar between clots from DVT and PE subjects. Electron microscopy revealed that plasma fibrin clots from PE subjects exhibited lower fiber density compared with those from DVT subjects. These data suggest that clot structure and functional properties differ between DVT and PE subjects and provide insights into mechanisms that may regulate embolization.

Abstract The muscle and cytoskeletal protein actin is released from cells as a consequence of cell death and interacts with components of the hemostatic and fibrinolytic systems, including platelets, plasmin, and fibrin. We report here that incorporation of actin filaments into fibrin clots changes their viscoelastic properties by increasing their shear modulus at low deforming stresses and by nearly eliminating their tendency to become more rigid with increasing deformation (ie, exhibit strain-hardening). The viscoelastic effects depended on the length of the actin filaments as shown by the effects of the plasma filament- severing protein, gelsolin. Binding of actin to fibrin clots also varied with actin filament length. The plasma actin-binding proteins gelsolin and vitamin D-binding protein reduced, but did not eliminate, the incorporation of actin in the clot. Fluorescence microscopy showed a direct association of rhodamine-labeled actin filaments with the fibrin network. Incubation of clots containing long actin filaments in solutions containing physiologic concentrations of gelsolin (2 mumol/L) released 60% of the actin trapped in the clot. Reduction of the actin content of a fibrin clot by incubation in a gelsolin-containing solution resulted in an increased rate of clot lysis. The ability of plasma gelsolin to shorten actin filaments may therefore be of physiologic and potentially of therapeutic importance insofar as gelsolin-mediated diffusion of actin from the clot may restore the clot's rheologic properties and render it more sensitive to the lytic action of plasmin.

Particulate matter (PM) as an important part of ambient air pollution has been associated with increased risks of cardiovascular diseases. Fibrin clot structure alteration is an emerging risk factor of many cardiovascular diseases, especially thrombosis. Therefore, the aim of this study was to investigate whether and how air particulate matter affects fibrin clot structure and endothelial cell behaviour. Turbidity assay, turbidity lysis assay and laser scanning confocal microscopy were used to analyse clots formed from normal pooled plasma or purified fibrinogen, in the presence of varying concentrations of PM. It was found that clots formed from plasma with higher concentrations of particles led to prolonged lysis time compared to control. No differences were seen for clots formed from fibrinogen. In a study of clots formed from plasma samples collected as part of a previous study on the effects of air pollution on deep vein thrombosis (DVT), alterations were observed in clots formed from plasma of DVT patients exposed to high levels of PM compared to those exposed to low levels, but the same differences were not observed in clots formed from plasma of control subjects. To investigate the potential role of venous endothelial cells in moderating clot structure following exposure to PM, human umbilical vein endothelial cells (HUVEC) were treated with PM for 24 hours and clots subsequently formed on the cells. Clots formed from plasma on the treated cells were altered compared to controls. RT-PCR and ELISA results showed increased gene expression of tissue factor (TF), protein expression of von Willebrand Factor (VWF) and plasminogen activation inhibitor-1 (PAI-1) and decreased thrombomodulin mRNA expression which were consistent with changes observed in clot structure. Engineered SiO2 nanoparticles caused denser clot structure in clots formed from normal pooled plasma. The gene expression of thrombomodulin was inhibited by SiO2 nanoparticles, but there were no significant difference in the TF mRNA expression between control and treated cells. Silica NPs caused increased concentrations of VWF, but not PAI-1 produced by endothelial cells. The results presented here show that PM can induce changes to clot structure and function, and that changes in gene expression induced in endothelial cells may be a mechanism by which a prothrombotic state is induced in response to PM exposure. Furthermore, some, but not all, similar changes were observed in clots and cells exposed to SiO2 nanoparticles, raising the possibility that such engineered nanoparticles may also have the potential to contribute to cardiovascular toxicity.

Statins are 3-hydroxy, 3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, which block the cholesterol biosynthetic pathway to lower total serum levels and LDL-cholesterol. The cholesterol pathway also provides a supply of isoprenoids (farnesyl and geranylgeranyl) for the prenylation of signaling molecules, which include the families of Ras and Rho small GTPases. Prenyl groups provide a membrane anchor that is essential for the correct membrane localisation and function of these proteins. Statins deplete cells of lipid geranylgeranyl diphosphate (GGPP) thereby inhibiting progression of the mevalonate pathway and prenylation of proteins. Two such proteins are Rab27b and Rap1, small GTPase proteins that are involved in the secretion of platelet granule and integrin activation. We hypothesise that statins can impair prenylation of Rab27b and Rap1a in platelets and thereby attenuate platelet function. The specific aims of the project were to analyse the impact of statins on the prenylation status of Rab27b and Rap1a in platelets. As Rab27b and Rap1a are known to be involved in secretion of platelet granules a secondary aim was to analyse the downstream effects of statins on this process following activation. Finally, we assessed the impact of treatment of platelets with statins on thrombus formation, stability and resistance to fibrinolysis. Platelets incubated with statins overnight were separated into cytosolic (aqueous) and membrane (detergent) components and visualised by Western blot. An accumulation of Rab27b and Rap1a was observed in the cytosolic compartments of statins treated platelets compared to untreated platelets, thus indicating indirect evidence that statins attenuate prenylation of Rab27b and Rap1a in platelets. The most effective statin in attenuating prenylation of Rab27b and Rap1a was atorvastatin (ATV). The inhibitory effect of statins on prenylation was recovered by GGPP, indicating that the mechanism of inhibition involved the mevalonate pathway. Release of ADP from platelet dense granules was significantly impeded following overnight treatment with ATV. In line with the inhibition of prenylation of Rab27b and Rap1a by ATV, addition of GGPP rescued the release of ADP from platelet dense granules. This suggests that attenuation of dense granules release by ATV occurs via interference in the mevalonate pathway and the inhibition of Rab27b prenylation. Furthermore, ATV significantly attenuates α-granules release in thrombin stimulated platelets, which was visualised as impaired accumulation of endogenous P-selectin, PAI-1 and fibrinogen on the activated membrane. Changes in the activation of α₁₁bβ₃ integrin on the stimulated platelet surface, observed as defective binding of exogenous fibrinogen and PAC-1, were also evident following treatment of platelets with ATV. In addition, ATV treatment of platelets reduced binding of CD41a, indicating that the copy number and activation of α₁₁bβ₃ integrin on stimulated platelets was significantly reduced. Statins were also found to significantly inhibit thrombin-induced platelet aggregation following incubation of platelets overnight with therapeutic concentrations of statins. Surprisingly GGPP did not rescue platelet aggregation indicating that different mechanisms are involved in inhibition of platelet responses by statins. Incubation of whole blood with ATV overnight significantly altered several haemostatic parameters. Using thromboelastography we demonstrated a delay in the coagulation time and clot formation time. Maximum clot firmness was also significantly reduced in the presence of statins compared to the control. The effect on clot firmness generally arises from platelet dysfunction and/or a change in fibrinogen concentration and function; the latter was ruled out using a Fibtem test, which shows no difference between treated and untreated whole blood. Similarly, formation of platelet-rich plasma clots was significantly delayed following pre-treatment with ATV overnight. These clots also exhibited lower maximal absorbances, which could represent differences in the fibrin network structure. In line with the reduction in fibrinogen binding defective clot retraction was also observed in platelet-rich plasma pre-treated with ATV overnight. Similar clot retraction results were observed with tirofiban and CytoD, suggesting that the inhibitory effect of ATV may involve modulation of α₁₁bβ₃ integrin activation. Platelet-rich plasma clots formed post-treatment with statins were visualised by confocal microscopy and revealed significant alterations in clot structure; observed as thinner fibrin fibres and fewer platelet aggregates. Additionally, we demonstrated that statins modulate clot stability and shorten time to lysis. Clots formed from platelet rich plasma that was subjected to incubation with ATV overnight revealed faster lysis by tPA compared to the absence of statin. These findings are also in agreement with the lysis of Chandler model thrombi formed from overnight incubated whole blood with ATV, which demonstrated faster lysis rate mediated by tPA. Furthermore, statins were shown to change the clot thrombodynamics as assessed by HemaCore analyser, which shows that stains implicate both clot growth in response to TF-coated comb and spontaneous clot lysis by tPA. In conclusion, statins directly inhibit Rab27b and Rap1a prenylation in platelets and down-regulated dense granules release. Inhibition of Rab27b and Rap1a prenylation, and dense granules release was recovered by GGPP, indicating that these effects are mediated through the mevalonate pathway. Impairment of platelet aggregation by statins resulted via multiple mechanisms as GGPP did not recovered the inhibition of aggregation by ATV. Statins also modulate fibrinogen binding, α-granules release, clot retraction and clot formation and stability in vitro. Together these results suggest that statins may directly attenuate the platelet response in vivo. The pleotropic effect of statins on platelets may contribute to the protective function of these class of drugs in cardiovascular diseases.

Hyperhomocysteinemia(Hhcys) is a condition that several epidemiological studies have shown to be associated with atherosclerosis and thrombosis, due to an elevation of plasma homocysteine levels. Plasma homocysteine(hcys) levels have a tendency to rise with age and changes in nutrition. Hcys can affect coagulation proteins, altering the formation of blood clots. The mechanism(s) by which hcys might cause modification of coagulation proteins "in vivo" is not understood. My hypothesis is that adult and juvenile animals could respond differently to chronic administration of hcys, and elevated plasma levels of hcys might lead to modification of fibrinogen "in vivo". Methodology: Six months old (juvenile, n=6) and 12 month old (adult, n=6) New Zealand White rabbits were divided into control (n=3) and homocysteine-treated (hcys-trt) (n=3) groups and injected for seven weeks; afterwards, they were given a bolus injection of hcys. Blood was drawn to evaluate plasma clearance of hcys. At the end, rabbits were exsanguinated by cardiac puncture and blood was collected for coagulation studies. Results: Juvenile hcys-trt rabbits adapted to chronic administration of hcys, however, adult hcys-trt rabbits developed Hhcys. Adult hcys-trt rabbits had higher levels of malonaldehyde in liver tissue, which is evidence of oxidative stress. Juvenile hcys-trt rabbits had similar malonaldehyde levels as juvenile control rabbits. Plasma elimination of hcys was impaired in adult hcys-trt rabbits. Adult hcys-trt rabbits had increased fibrinogen levels, longer reptilase times, and shorter thrombin clotting times versus adult control rabbits. Clots formed from purified fibrinogen obtained from hcys-trt rabbits lysed slower than comparable clots formed from control rabbits purified fibrinogen. Some congenital dysfibrinogenemias have clots that are abnormally resistant to fibrinolysis due to alterations in fibrinogen structure, and lead to recurrent thrombosis. Clotting results for adult hcys-trt rabbits suggest that hyperhomocysteinemia leads to a similar acquired dysfibrinogenmia. Therefore, the prolonged reptilase times and formation of clots that are abnormally resistant to fibrinolysis could directly contribute to increased risk of thrombosis in hyperhomocysteinemia.

Hematomas (blood clots) are known important factors in early stages of fracture repair. This project characterized the differences in hematomas between normal healing and delayed healing in bone defects. It was found that manipulation of the hematoma structure can significantly improve bone healing and result in significantly increased bone formation. This project will lead to the development of innovative treatment of large bone defects, fractures, and bone non-union.

La formation du caillot de fibrine, processus clé de la coagulation sanguine, implique la polymérisation des monomères de fibrine en un réseau de fibres. Ce réseau contrôle les propriétés mécaniques du caillot et constitue le squelette sur lequel se base la cicatrisation. Si l’influence des conditions de réaction (pH, concentration, …) est bien connue, le rôle de la composition du fibrinogène sur la structure de la fibrine est inexploré. Cet aspect pourrait être important pour les pathologies cardiovasculaires qui présentent toutes une structure de fibrine anormale.Nous avons étudié la relation entre la composition de plusieurs fibrinogènes et les propriétés structurelles nano- et micro-métriques ainsi que la mécanique des caillots de fibrine. La composition en protéines co-purifiées de ces fibrinogènes a peu d’influence, alors que le profil de polydispersité contrôle la structure multi-échelle de la fibrine. Des mesures de diffusion des rayons x, de spectrophotométrie multi-longueur d’ondes et de microscopie confocale ont mis en évidence que les fibres provenant des fibrinogènes monodisperses sont quasi-cristallines, droites et rigides. Les fibres provenant de fibrinogènes polydisperses sont, elles, beaucoup moins organisées, courbées, avec un module de rigidité faible. Enfin, les propriétés mécaniques de la fibrine ont montré que la réponse des caillots aux déformations, aussi que les scenarios de rupture, sont directement liés à sa structure et donc significativement dépendants du profil de polydispersité des fibrinogènes. Ces résultats ouvrent de nouvelles perspectives dans plusieurs domaines, que ce soit pour l’utilisation optimale des fibrinogènes pour les dysfibrinogénémies et hémorragies, mais également pour la reconstruction tissulaire, ainsi que la compréhension du lien entre la structure anormale des caillots et les maladies cardiovasculaires
Fibrin clot formation is one of the major processes leading to blood clotting. It involves the polymerization of fibrin monomers into a network of fibrin fibres. This network controls the mechanical properties of the clot and serves as a skeleton for wound healing. Environmental factors (pH, concentration, …) have been proved to influence polymerization, however the role of fibrinogen composition on the structure of fibrin remains unexplored. This aspect might be important for the case of cardiovascular pathologies, which present abnormal fibrin structures.We have determined the relation between different sources of fibrinogen with the nano- and micro-metric structural and mechanical properties of fibrin clots. The composition in co-purified proteins of the fibrinogens has no significant importance, however the polydispersity profile controls the multiscale properties of fibrin. Indeed, x-ray scattering, multi-wavelength spectrophotometry and confocal microscopy measurements have proved that fibres from monodisperse fibrinogens are quasi-crystalline, straight and rigid. Fibres from polydisperse fibrinogens are less organised, curbed and less rigid. Finally, the mechanical properties of fibrin showed that the response of clots to deformation, as well as the scenarios of rupture are closely related to the structure, and consequently related to the profiles of polydispersity. This opens outstanding perspectives in many fields such the optimisation of fibrinogen’s use on dysfibrinogenemias or haemorrhages, tissue regeneration or the understanding between the abnormal structure of clots and cardiovascular diseases

Atrial fibrillation (AF) is the most common arrhythmia and is closely associated with chronic kidney disease. The mainstay pharmacological agent to prevent AF-related stroke and thromboembolism is the use of oral anticoagulants, but may result in an increased risk of haemorrhage. Therefore, this MD research thesis is a comprehensive study of the changes in thrombogenesis and fibrin clot structure related to AF and CKD, as well as the potential impact of exposure to different classes of oral anticoagulant.

Le rôle physiologique du caillot est d’éviter un épanchement excessif de sang en présence d’une brèche vasculaire. Une fois cette fonction remplie, il doit pouvoir être facilement détruit, afin qu’il ne passe pas dans le système veineux et ne gêne la circulation sanguine. La formation d’un caillot de fibrine et sa lyse, processus clés de l’hémostase, impliquent à la fois la polymérisation des monomères de fibrinogène en un réseau de fibres de fibrine, et la résorption du réseau de fibres de fibrine constitué. Bien que ce réseau contrôle l’ensemble des propriétés physiques et mécaniques du caillot, sa structure aux échelles inférieures au micron est très mal caractérisée. Le principal verrou à la caractérisation physique du caillot en environnement clinique est l’absence de méthode de mesure quantitative, fiable, sensible et reproductible. Il est donc nécessaire de produire une méthode de mesure adéquate, couplée à un système de mesure sensible. Nous avons démontré dans ce travail, grâce à notre méthode utilisant plusieurs longueurs d’onde, que l’analyse du spectre de lumière visible transmis à travers un caillot permet de déterminer simultanément, quantitativement et en conditions quasi-physiologiques, plusieurs paramètres essentiels de structure du caillot de fibrine, à savoir le nombre de protofibrilles par fibre de fibrine, le rayon et la densité de ces fibres, ainsi que les temps de formation et de lyse du caillot. Cette technique a été validée via les résultats avec des CV inférieurs dans l’ensemble à 6% sous plusieurs conditions de tests et différents profils plasmatiques : normaux, hypo/hyper coagulants et hypo/hyper fibrinolytiques, attestant de la robustesse et de la fiabilité de la technique de mesure aussi bien pour le suivi de la coagulation que de la lyse. Cette méthode de spectrophotométrie a pu être implantée sur un automate modifié à des fins de diagnostic et à vocation hospitalière pour des plasmas de patients présentant des troubles de l’hémostase. Les informations cliniques et intérêts attendus de ce nouveau test, concernent à la fois la qualité du réseau de fibrine, sa lyse accélérée ou sa résistance à la fibrinolyse ainsi que la résultante de la balance coagulo-lytique
The physiological role of the clot is to avoid excessive bleeding in the presence of a vascular breach. Once this function is filled, the clot must be able to be easily destroyed, so that it is not transported in the venous system and does not hamper blood circulation. The formation of a fibrin clot and its lysis are key processes of hemostasis, implying simultaneously the polymerization of the fibrinogen monomers in a fibrin fibers network, and the destruction of this constituted network.Although this network controls the physical and mechanical properties of the clot, its structure at scales smaller than the micron is poorly characterized. The main problem in the physical characterization of clot in clinical settings is the current absence of a quantitative, sensitive and reproducible measurement method.We demonstrated in this work, thanks to our method using several wavelengths, that the analysis of the visible spectra of light transmitted through a clot allows to determine simultaneously, quantitatively and in quasi-physiological conditions, several essential parameters of structure of the fibrin clot, namely the number of protofibrils per fibrin fibers, the radius and the density of fibers, and various times of clotting and lysis of the clot. This method was validated by the results with CV inferior to 6 % under all test conditions and various plasmatic profiles: normal, hypo / hyper coagulant and hypo / hyper fibrinolytic. This demonstrates the robustness and reliability of the measurement method when measuring both clotting and clot lysis.This spectrophotometric method was implemented on a modified automaton dedicated to diagnosis of patients presenting hemostatic disorders. The clinical information and the interests expected from this new test concern at the same time the quality of the fibrin network, its accelerated lysis or its resistance to fibrinolysis, and the resultant of the coagulo-lytic balance

Physiologiquement, le caillot sanguin a pour fonction l’arrêt d’un saignement consécutif à une brèche vasculaire. Dans un premier temps, ce sont les plaquettes sanguines qui stoppent l’épanchement sanguin, rapidement soutenues par la formation d’un réseau de fibres de fibrine qui consolide et confère au caillot les propriétés nécessaires pour résister à la pression sanguine et à la fibrinolyse. Le fibrinogène est l’élément de base du réseau de fibrine. Lors d’une brèche vasculaire, la libération de facteur tissulaire entraine le déclenchement de la cascade de coagulation qui aboutit à la transformation du fibrinogène en monomères de fibrine par l’action de la thrombine. Ceux-ci s’agrègent longitudinalement pour former des protofibrilles, puis latéralement pour former un réseau de fibres de fibrine.A ce jour, de nombreuses étapes de formation du caillot ont été décrites en détail dans la littérature, cependant les mécanismes et les forces motrices de l’agrégation latérale des protofibrilles sont encore mal compris.Lors de ce travail, nous avons étudié différents profils de coagulation : de l’hypo-coagulant à l’hypercoagulant, en passant par le profil normal et en utilisant un panel varié de techniques : génération de thrombine, génération de plasmine, Fibrinographie, Fibrinographie en mode « fibrinolyse », microscopie confocale, thromboélastométrie et diffraction des rayons X aux petits angles.Nous avons mis en évidence la relation entre la quantité de thrombine présente lors de la formation d’un caillot et la structure de celui-ci. En effet, Plus il y a de thrombine, plus le nombre de protofibrilles par fibre est faible et plus le nombre de fibres est important. De plus, nous avons corrélé le temps d’initiation de l’agrégation latérale des fibres en Fibrinographie avec l’initiation de la génération de plasmine. Nous avons ainsi mis en évidence la production d’une structure du caillot de fibrine anormale en présence de dabigatran, grâce à l’utilisation combinée de la microscopie confocale et de la Fibrinographie.Cette analyse multimodale de la structure du caillot dans différentes conditions apporte des informations supplémentaires à la communauté scientifique, pour permettre de mieux comprendre les mécanismes de formation des caillots de fibrine
Physiologically, the blood function of the clot is to stop bleeding following a vascular breach. Initially, platelets stop blood flow, quickly supported by the formation of a fibrin fibers network that strengthens and gives properties to resist the blood pressure and fibrinolysis. Fibrinogen is the basic element of the fibrin network. During a vascular breach, the release of tissue factor triggers the coagulation cascade that results in the conversion of fibrinogen to fibrin monomers by the action of thrombin. These aggregate longitudinally to form protofibrils, then laterally to form a network of fibrin fibers.To date, many stages of the clot formation have been described in detail in the literature, however the mechanisms and driving forces of the lateral aggregation of protofibrils are still poorly understood.During this work, we studied different coagulation profiles: from hypo-coagulant to hyper-coagulant, through the normal profile and using a varied range of techniques: thrombin generation, plasmin generation, Fibrinography, Fibrinography in "fibrinolysis" mode, confocal microscopy, thromboelastometry and X-ray diffraction at small angles.We have highlighted the relationship between the amount of thrombin present during clot formation and the clot structure. Indeed, the more thrombin there is, the lower the protofibrils number per fiber and the greater the number of fibers. In addition, we correlated the initiation time of lateral fibers aggregation in Fibrinography with the initiation of plasmin generation. We have thus demonstrated the production of an abnormal fibrin clot structure in the presence of dabigatran, thanks to the combined use of confocal microscopy and Fibrinography.This multimodal analysis of the clot structure under different conditions provides additional information to the scientific community to better understand the mechanisms of fibrin clot formation

The platelet is a small (2–4 µm in diameter), discoid, anucleate cell that circulates in the blood. In health, it plays a vital role in haemostasis, and in disease it contributes to disorders of bleeding and thrombosis. Platelets are produced from the surface of megakaryocytes in the bone marrow, under tight homeostatic control regulated by the cytokine thrombopoietin. Platelets have a lifespan of approximately 7–10 days, and usually circulate in the blood stream in a quiescent state. Intact, undamaged vessel walls help to maintain platelets in this inactive state by releasing nitric oxide, which acts both to dilate the vessel wall and to inhibit platelet adhesion, activation, and aggregation. After trauma to the blood vessel wall, platelets are activated and, acting in concert with the endothelium and coagulation factors, form a stable clot. This chapter addresses platelet structure and function, and the response of platelets to vessel injury.

Abstract This chapter focuses on greenhouses and protected cropping structures. Topics covered are glasshouses and plastic greenhouses, closed and semi-closed greenhouse structures, passive solar greenhouses, sustainable greenhouse design, cladding materials, screen houses, net houses, shade houses, rain covers and other structures, screen and shade nets, low tunnels and high tunnels, hot beds and cold frames greenhouses, floating mulches, row covers, cloche covers, direct covers and frost cloth, greenhouse site planning, windbreaks, outdoor hydroponic systems, and controlled-environment agriculture.

Abstract This chapter focuses on greenhouses and protected cropping structures. Topics covered are glasshouses and plastic greenhouses, closed and semi-closed greenhouse structures, passive solar greenhouses, sustainable greenhouse design, cladding materials, screen houses, net houses, shade houses, rain covers and other structures, screen and shade nets, low tunnels and high tunnels, hot beds and cold frames greenhouses, floating mulches, row covers, cloche covers, direct covers and frost cloth, greenhouse site planning, windbreaks, outdoor hydroponic systems, and controlled-environment agriculture.

Mechanical thrombectomy devices, such as retrievers or aspiration catheters, have recently received approval from the FDA for the treatment of acute ischemic stroke. There is growing interest in endovascular recanalization procedures due to mounting evidence of favorable clinical outcomes. Several attempts have been made to establish dedicated clot models for in-vitro or in-vivo simulation of thromboembolism [1,2]. However, little is known about the mechanical and structural similarities between experimental clots and human sources of emboli that cause stroke. The goal of this study is to compare the structure and compression behavior of the possible sources of the cerebral emboli extracted from patients and model clots produced in-vitro using human, porcine and bovine donors.

When a blood vessel is damaged, platelets aggregate together to form a thrombus that seals vascular leakage and promotes healing. The process of thrombosis involves activation of platelets by soluble ligands, adhesion to the wound site, change in shape from a discoidal to a stellate structure, and the attachment of additional platelets onto the nascent clot through adhesive ligand bridges. Once formed, a clot will undergo a retraction in volume that packs platelets together and allows blood flow to recommence. Much attention has been paid to the early phases of thrombosis — adhesion and aggregation — but the process of clot retraction is vital to stabilizing the clot.

Aα 16 Arg→His substitutions are common forms of congenital dysfibrinogenemias. Clinical manifestations range from asymptomatic to moderate hemorrhagic tendencies. Biochemical characterization of one such heterozygotic individual (Fibrinogen Louisville, Galanakis, etal. Ann NY Acad Sci 408:644,1983) indicated that only homodimeric fibrinogen molecules (i.e., containing either normal or abnormal Aα chains) were present. We isolated fibrinogen from the plasma of a 23 year old patient with a history of easy bruising and several recent moderate to severe bleeding episodes. Coagulability with reptilase was 677 (65-70%; n=5) whereas with thrombin (Ha) it approached 100%, depending directly upon the time of incubation with enzyme. HPLC analysis of Ila-induced fibrinopeptide release demonstrated the presence of an abnormal A-peptide (A*), amounting to 50% of the total, which was released more slowly than the normal A-peptide (A). Amino acid analysis of A* demonstrated the absence of Arg and the presence of His. Carboxypeptidase digestion confirmed the structure of A* as Aα 16 Arg-→ His. The clot and the soluble clot liquor resulting from reptilase treatment were separated and each was then further treated with Ilato release A*. HPLC analysis indicated that 31% of the total A* present in the sample was associated with the reptilase clot and 697 remained in the clot liquor. This distribution of A* suggests that Fibrinogen Birmingham, unlike Fibrinogen Louisville, contains heterodimeric molecules that are incorporated into the reptilase clottable fraction. This finding is consistent with a process of random hepatic assembly of dimeric fibrinogen molecules in a heterozygotic individual.

α chain polymerisation during clot formation is accelerated in the presence of erythrocytes. This effect is abrogated if the erythrocytes are obtained from patients with various haemoglo-binopathies. Enzyme digests of the α polymers produced in the presence or absence of erythrocytes were prepared to further define any differences between them.Clots were produced from citrate/EACA plasma samples or plasma/erythrocyte mixtures by the addition of thrombin and calcium. After five hours, clots were washed iii 8 M urea until traces of haemoglobin were removed. After reduction and alkylation clots were dissolved in 0.5% SDS. a polymers were purified on sephacryl S-300 and protein concentrations were adjusted to 0.5 mg/ml. These were digested with S. Aureus V8 protease (150 mg/ml), papain (50 mg/ml) or chymotrypsin (100 mg/ml) at 37°C at sequential time intervals.After the addition of 2% SDS samples were analysed on 15% SDS polyacrylamide gels.In all cases digestion of a polymers from clots formed in the absence of erythrocytes took place more rapidly and contained peptide bands not apparent in other digests.The observations suggest that α polymers formed in the presence or absence of erythrocytes exhibit differing kinetics of response to proteolytic cleavage and indicate that erythrocytes may influence the primary and/or quartenary structure of the polymers studied.

Cette étude porte sur une des innovations contemporaines liées à l’économie de la publication scientifique : les accords dits transformants, un objet relativement circonscrit au sein des relations entre consortiums de bibliothèques et éditeurs scientifiques, et temporellement situé entre 2015 et 2020. Ce type d’accords a pour objectif affiché d’organiser la transition du modèle traditionnel de l’abonnement à des revues (souvent proposées par regroupements thématiques ou collections) vers celui de l’accès ouvert en opérant une réaffectation des budgets qui y sont consacrés. Notre travail d’analyse sociologique constitue une première étude systématique de cet objet, fondée sur la recension de 197 accords. Le corpus ainsi constitué inclut des accords caractérisés par la coprésence d’une composante d’abonnement et d’une composante de publication en accès ouvert, même minimale (« jetons » de publication offerts, réduction sur les APC...). En conséquence, ont été exclus de l’analyse les accords portant uniquement sur du financement centralisé de publication en accès ouvert, que ce soit avec des éditeurs ne proposant que des revues avec paiement par l’auteur (PLOS, Frontiers, MDPI...) ou des éditeurs dont une partie du catalogue est constitué de revues en accès ouvert. L’accord le plus ancien de notre corpus a été signé en 2010, les plus récents en 2020 – les accords ne commençant qu’en 2021, même annoncés au cours de l’étude, n’ont pas été retenus. Plusieurs résultats se dégagent de notre analyse. Tout d’abord, on note une grande diversité des acteurs impliqués avec 22 pays et 39 éditeurs, même si certains consortiums (Pays-Bas, Suède, Autriche, Allemagne) et éditeurs (CUP, Elsevier, RSC, Springer) en ont signé beaucoup plus que d’autres. Ensuite, la durée des accords, comprise entre une et six années, révèle une distribution très inégalitaire, avec plus de la moitié des accords (103) signés pour 3 ans, ainsi qu’une faible proportion pour 4 ans ou plus (22 accords). Enfin, en dépit d’appels répétés à la transparence, moins de la moitié des accords (96) ont un texte accessible au moment de cette étude, sans qu’on puisse observer une tendance récente à une plus grande disponibilité. L’analyse montre également des degrés d’ouverture très variables, allant d’une simple information sur le répertoire ESAC en passant par la mise à disposition d’un format annotable jusqu’à l’attribution d’un DOI et d’une licence de réutilisation (CC-BY), en incluant le détail des sommes monétaires. Parmi les 96 accords disponibles, dont 47 signés en 2020, 62 ont fait l’objet d’une analyse en profondeur. C’est à notre connaissance la première analyse à cette échelle, sur un type de matériel non seulement inédit, mais qui était auparavant soumis à des clauses de confidentialité. Fondée sur une lecture minutieuse, l’étude décrit de manière fine leurs propriétés, depuis la matérialité du document jusqu’aux formules financières, en passant par leur morphologie et l’ensemble des droits et devoirs des parties. Les contenus des accords sont donc analysés comme une collection dont nous cherchons à déterminer les points communs et les variations, à travers des codages explicites sur certaines de leurs caractéristiques. L’étude pointe également des incertitudes, et notamment leur caractère « transitionnel », qui demeure fortement discuté. D’un point de vue morphologique, les accords montrent une grande diversité en matière de taille (de 7 à 488 pages) et de structure. Néanmoins, par définition, ils articulent tous deux objets essentiels : d’une part, les conditions de réalisation d’une lecture d’articles de revues, sous forme d’abonnement, mêlant des préoccupations d’accès et de sécurité ; d’autre part, les modalités de publication en accès ouvert, articulant la gestion d’un nouveau type de workflow à toute une série d’options possibles. Parmi ces options, mentionnons notamment le périmètre des revues considérées (hybrides et/ou accès ouvert), les licences disponibles, le degré d’obligation de cette publication, les auteurs éligibles ou le volume d’articles publiables. L’un des résultats les plus importants de cette analyse approfondie est la mise au jour d’un découplage presque complet, au sein même des accords, entre l’objet abonnement et l’objet publication. Bien entendu, l’abonnement est systématiquement configuré dans un monde fermé, soumis à paiement qui déclenche des séries d’identification des circulations légitimes tant du contenu informationnel que des usagers. Il insiste notamment sur les interdictions de réutilisation ou même de copie des articles scientifiques. À l’opposé, la publication en accès ouvert est attachée à un monde régi par l’accès gratuit au contenu, ce qui induit des préoccupations de gestion du workflow et des modalités d’accessibilité. De plus, les différents éléments constitutifs de ces objets contractuels ne sont pas couplés : d’un côté, les lecteurs sont constitués de l’ensemble des membres des institutions abonnées, de l’autre, seuls les auteurs correspondants (« corresponding authors ») sont concernés ; les listes de revues accessibles à la lecture et celles réservées à la publication en accès ouvert sont le plus souvent distinctes ; les workflows ont des objectifs et des organisations matérielles totalement différentes, etc. L’articulation entre les deux objets contractuels relève uniquement d’une formule de distribution financière qui, outre des combinaisons particulières entre l’un et l’autre, permet d’attribuer des étiquettes distinctes aux accords (offset agreement, publish & read, read & publish, read & free articles, read & discount). Au-delà de cette distribution, l’étude des arrangements financiers montre une gamme de dispositions allant d’une prévisibilité budgétaire totale, donc identique aux accords d’abonnement antérieurs, à une incertitude sur le volume de publication ou sur le montant définitif des sommes échangées. Les modalités concrètes de calcul des montants associés à la publication en accès ouvert sont relativement variées. S’il existe effectivement des formules récurrentes (volume d’articles multiplié par un prix individuel, reprise de la moyenne des sommes totales d’APC des années précédentes...), le calcul des sommes en jeu est toujours le résultat d’une négociation singulière entre un consortium et un éditeur scientifique, et aboutit parfois à des formules originales et complexes. À ce titre, l’espace des possibles en matière de formules financières n’est jamais totalement clos. Par ailleurs, la volonté des consortiums d’opérer une « transformation » de leurs accords vers la publication à coût constant renvoie à des définitions diversifiées du « coût » (inclusion ou non des dépenses d’APC préexistantes) et de la constance (admission ou pas d’une « inflation » à 2 ou 3%). De plus, nous n’avons observé aucune disposition contractuelle permettant d’anticiper les sommes en jeu au-delà de l’horizon temporel de l’accord courant. La grande diversité des accords provient d’une part des conditions initiales des relations entre consortiums et éditeurs scientifiques – les sommes dépensées en abonnement étant le point de départ des nouveaux accords –, d’autre part des objectifs de chaque partie. Même si cette étude excluait volontairement les négociations, les accords portent des traces de ces objectifs. Ainsi, de nombreux accords sont de nature explicitement expérimentale, quand certains visent un contrôle budgétaire strict, ou d’autres ambitionnent, dans la période plus récente, la publication du plus grand nombre possible d’articles en accès ouvert. C’est dans ce dernier cas qu’on touche à l’ambiguïté des attentes générales sur les accords transformants. En effet, pour les consortiums, la dimension « transformante » consiste essentiellement à transférer les sommes traditionnellement allouées à l’abonnement vers la publication en accès ouvert. Mais l’objectif n’est jamais de transformer le modèle économique des revues, c'est-à-dire de faire basculer des revues sous abonnement ou hybrides en revues entièrement en accès ouvert. D’ailleurs, aucune clause ne vise une telle fin – à l’exception du modèle d’accord proposé par l’éditeur ACM. Du côté des éditeurs, et notamment de Springer, le caractère cumulatif des accords nationaux passés vise à projeter un monde de la publication où l’accès ouvert devient de fait quantitativement très dominant, sans pour autant modifier de manière pérenne le modèle économique de leurs revues. Notre étude montre que les accords transformants actuels ne permettent pas d’assurer de manière durable une transition de l’économie de la publication vers l’accès ouvert, dans la mesure où ils n’offrent pas de garantie sur le contrôle des dépenses ni sur la pérennité de l’ouverture des contenus. L’avenir des relations entre consortium et éditeur demeure largement indéterminé.Cette étude porte sur une des innovations contemporaines liées à l’économie de la publication scientifique : les accords dits transformants, un objet relativement circonscrit au sein des relations entre consortiums de bibliothèques et éditeurs scientifiques, et temporellement situé entre 2015 et 2020. Ce type d’accords a pour objectif affiché d’organiser la transition du modèle traditionnel de l’abonnement à des revues (souvent proposées par regroupements thématiques ou collections) vers celui de l’accès ouvert en opérant une réaffectation des budgets qui y sont consacrés. Notre travail d’analyse sociologique constitue une première étude systématique de cet objet, fondée sur la recension de 197 accords. Le corpus ainsi constitué inclut des accords caractérisés par la coprésence d’une composante d’abonnement et d’une composante de publication en accès ouvert, même minimale (« jetons » de publication offerts, réduction sur les APC...). En conséquence, ont été exclus de l’analyse les accords portant uniquement sur du financement centralisé de publication en accès ouvert, que ce soit avec des éditeurs ne proposant que des revues avec paiement par l’auteur (PLOS, Frontiers, MDPI...) ou des éditeurs dont une partie du catalogue est constitué de revues en accès ouvert. L’accord le plus ancien de notre corpus a été signé en 2010, les plus récents en 2020 – les accords ne commençant qu’en 2021, même annoncés au cours de l’étude, n’ont pas été retenus. Plusieurs résultats se dégagent de notre analyse. Tout d’abord, on note une grande diversité des acteurs impliqués avec 22 pays et 39 éditeurs, même si certains consortiums (Pays-Bas, Suède, Autriche, Allemagne) et éditeurs (CUP, Elsevier, RSC, Springer) en ont signé beaucoup plus que d’autres. Ensuite, la durée des accords, comprise entre une et six années, révèle une distribution très inégalitaire, avec plus de la moitié des accords (103) signés pour 3 ans, ainsi qu’une faible proportion pour 4 ans ou plus (22 accords). Enfin, en dépit d’appels répétés à la transparence, moins de la moitié des accords (96) ont un texte accessible au moment de cette étude, sans qu’on puisse observer une tendance récente à une plus grande disponibilité. L’analyse montre également des degrés d’ouverture très variables, allant d’une simple information sur le répertoire ESAC en passant par la mise à disposition d’un format annotable jusqu’à l’attribution d’un DOI et d’une licence de réutilisation (CC-BY), en incluant le détail des sommes monétaires. Parmi les 96 accords disponibles, dont 47 signés en 2020, 62 ont fait l’objet d’une analyse en profondeur. C’est à notre connaissance la première analyse à cette échelle, sur un type de matériel non seulement inédit, mais qui était auparavant soumis à des clauses de confidentialité. Fondée sur une lecture minutieuse, l’étude décrit de manière fine leurs propriétés, depuis la matérialité du document jusqu’aux formules financières, en passant par leur morphologie et l’ensemble des droits et devoirs des parties. Les contenus des accords sont donc analysés comme une collection dont nous cherchons à déterminer les points communs et les variations, à travers des codages explicites sur certaines de leurs caractéristiques. L’étude pointe également des incertitudes, et notamment leur caractère « transitionnel », qui demeure fortement discuté. D’un point de vue morphologique, les accords montrent une grande diversité en matière de taille (de 7 à 488 pages) et de structure. Néanmoins, par définition, ils articulent tous deux objets essentiels : d’une part, les conditions de réalisation d’une lecture d’articles de revues, sous forme d’abonnement, mêlant des préoccupations d’accès et de sécurité ; d’autre part, les modalités de publication en accès ouvert, articulant la gestion d’un nouveau type de workflow à toute une série d’options possibles. Parmi ces options, mentionnons notamment le périmètre des revues considérées (hybrides et/ou accès ouvert), les licences disponibles, le degré d’obligation de cette publication, les auteurs éligibles ou le volume d’articles publiables. L’un des résultats les plus importants de cette analyse approfondie est la mise au jour d’un découplage presque complet, au sein même des accords, entre l’objet abonnement et l’objet publication. Bien entendu, l’abonnement est systématiquement configuré dans un monde fermé, soumis à paiement qui déclenche des séries d’identification des circulations légitimes tant du contenu informationnel que des usagers. Il insiste notamment sur les interdictions de réutilisation ou même de copie des articles scientifiques. À l’opposé, la publication en accès ouvert est attachée à un monde régi par l’accès gratuit au contenu, ce qui induit des préoccupations de gestion du workflow et des modalités d’accessibilité. De plus, les différents éléments constitutifs de ces objets contractuels ne sont pas couplés : d’un côté, les lecteurs sont constitués de l’ensemble des membres des institutions abonnées, de l’autre, seuls les auteurs correspondants (« corresponding authors ») sont concernés ; les listes de revues accessibles à la lecture et celles réservées à la publication en accès ouvert sont le plus souvent distinctes ; les workflows ont des objectifs et des organisations matérielles totalement différentes, etc. L’articulation entre les deux objets contractuels relève uniquement d’une formule de distribution financière qui, outre des combinaisons particulières entre l’un et l’autre, permet d’attribuer des étiquettes distinctes aux accords (offset agreement, publish & read, read & publish, read & free articles, read & discount). Au-delà de cette distribution, l’étude des arrangements financiers montre une gamme de dispositions allant d’une prévisibilité budgétaire totale, donc identique aux accords d’abonnement antérieurs, à une incertitude sur le volume de publication ou sur le montant définitif des sommes échangées. Les modalités concrètes de calcul des montants associés à la publication en accès ouvert sont relativement variées. S’il existe effectivement des formules récurrentes (volume d’articles multiplié par un prix individuel, reprise de la moyenne des sommes totales d’APC des années précédentes...), le calcul des sommes en jeu est toujours le résultat d’une négociation singulière entre un consortium et un éditeur scientifique, et aboutit parfois à des formules originales et complexes. À ce titre, l’espace des possibles en matière de formules financières n’est jamais totalement clos. Par ailleurs, la volonté des consortiums d’opérer une « transformation » de leurs accords vers la publication à coût constant renvoie à des définitions diversifiées du « coût » (inclusion ou non des dépenses d’APC préexistantes) et de la constance (admission ou pas d’une « inflation » à 2 ou 3%). De plus, nous n’avons observé aucune disposition contractuelle permettant d’anticiper les sommes en jeu au-delà de l’horizon temporel de l’accord courant. La grande diversité des accords provient d’une part des conditions initiales des relations entre consortiums et éditeurs scientifiques – les sommes dépensées en abonnement étant le point de départ des nouveaux accords –, d’autre part des objectifs de chaque partie. Même si cette étude excluait volontairement les négociations, les accords portent des traces de ces objectifs. Ainsi, de nombreux accords sont de nature explicitement expérimentale, quand certains visent un contrôle budgétaire strict, ou d’autres ambitionnent, dans la période plus récente, la publication du plus grand nombre possible d’articles en accès ouvert. C’est dans ce dernier cas qu’on touche à l’ambiguïté des attentes générales sur les accords transformants. En effet, pour les consortiums, la dimension « transformante » consiste essentiellement à transférer les sommes traditionnellement allouées à l’abonnement vers la publication en accès ouvert. Mais l’objectif n’est jamais de transformer le modèle économique des revues, c'est-à-dire de faire basculer des revues sous abonnement ou hybrides en revues entièrement en accès ouvert. D’ailleurs, aucune clause ne vise une telle fin – à l’exception du modèle d’accord proposé par l’éditeur ACM. Du côté des éditeurs, et notamment de Springer, le caractère cumulatif des accords nationaux passés vise à projeter un monde de la publication où l’accès ouvert devient de fait quantitativement très dominant, sans pour autant modifier de manière pérenne le modèle économique de leurs revues. Notre étude montre que les accords transformants actuels ne permettent pas d’assurer de manière durable une transition de l’économie de la publication vers l’accès ouvert, dans la mesure où ils n’offrent pas de garantie sur le contrôle des dépenses ni sur la pérennité de l’ouverture des contenus. L’avenir des relations entre consortium et éditeur demeure largement indéterminé.