Transcriptomic Analysis of Extracellular Vesicles in the Search for Novel Plasma and Thrombus Biomarkers of Ischemic Stroke Etiologies.

Accurate etiologic diagnosis provides an appropriate secondary prevention and better prognosis in ischemic stroke (IS) patients; still, 45% of IS are cryptogenic, urging us to enhance diagnostic precision. We have studied the transcriptomic content of plasma extracellular vesicles (EVs) (n = 21) to identify potential biomarkers of IS etiologies. The proteins encoded by the selected genes were measured in the sera of IS patients (n = 114) and in hypertensive patients with (n = 78) and without atrial fibrillation (AF) (n = 20). IGFBP-2, the most promising candidate, was studied using immunohistochemistry in the IS thrombi (n = 23) and atrium of AF patients (n = 13). In vitro, the IGFBP-2 blockade was analyzed using thromboelastometry and endothelial cell cultures. We identified 745 differentially expressed genes among EVs of cardioembolic, atherothrombotic, and ESUS groups. From these, IGFBP-2 (cutoff > 247.6 ng/mL) emerged as a potential circulating biomarker of embolic IS [OR = 8.70 (1.84-41.13) p = 0.003], which was increased in patients with AF vs. controls (p < 0.001) and was augmented in cardioembolic vs. atherothrombotic thrombi (p < 0.01). Ex vivo, the blockage of IGFBP-2 reduced clot firmness (p < 0.01) and lysis time (p < 0.001) and in vitro, diminished endothelial permeability (p < 0.05) and transmigration (p = 0.06). IGFBP-2 could be a biomarker of embolic IS and a new therapeutic target involved in clot formation and endothelial dysfunction.

Role of factor XIII in ischemic stroke: a key molecule promoting thrombus stabilization and resistance to lysis.

BACKGROUND: Active coagulation factor XIII (FXIII) catalyzing crosslinking of fibrin and other hemostatic factors plays a key role in clot stability and lysis. OBJECTIVES: To evaluate the effect of FXIII inhibition in a mouse model of ischemic stroke (IS) and the role of activated FXIII (FXIIIa) in clot formation and lysis in patients with IS. METHODS: A ferric chloride IS murine model was performed before and after administration of a FXIIIa inhibitor (FXIIIinh). Thromboelastometry in human and mice blood was used to evaluate thrombus stiffness and lysis with FXIIIinh. FXIIIa-dependent fibrin crosslinking and lysis with fibrinolytic drugs (tissue plasminogen activator and tenecteplase) were studied on fibrin plates and on thrombi and clotted plasma of patients with IS. Finally, circulating and thrombus FXIIIa were measured in 85 patients with IS. RESULTS: FXIIIinh administration before stroke induction reduced infarct size, α2-antiplasmin (α2AP) crosslinking, and local microthrombosis, improving motor coordination and fibrinolysis without intracranial bleeds (24 hours). Interestingly, FXIII blockade after stroke also reduced brain damage and neurologic deficit. Thromboelastometry in human/mice blood with FXIIIinh showed delayed clot formation, reduced clot firmness, and shortened tissue plasminogen activator lysis time. FXIIIa fibrin crosslinking increased fibrin density and lysis resistance, which increased further after α2AP addition. FXIIIinh enhanced ex vivo lysis in stroke thrombi and fibrin plates. In patients with IS, thrombus FXIII and α2AP were associated with inflammatory and hemostatic components, and plasma FXIIIa correlated with thrombus α2AP and fibrin. CONCLUSION: Our results suggest a key role of FXIIIa in thrombus stabilization, α2AP crosslinking, and lysis resistance, with a protective effect of FXIIIinh in an IS experimental model.

Loss of the matrix metalloproteinase-10 causes premature features of aging in satellite cells.

Aged muscles accumulate satellite cells with a striking decline response to damage. Although intrinsic defects in satellite cells themselves are the major contributors to aging-associated stem cell dysfunction, increasing evidence suggests that changes in the muscle-stem cell local microenvironment also contribute to aging. Here, we demonstrate that loss of the matrix metalloproteinase-10 (MMP-10) in young mice alters the composition of the muscle extracellular matrix (ECM), and specifically disrupts the extracellular matrix of the satellite cell niche. This situation causes premature features of aging in the satellite cells, contributing to their functional decline and a predisposition to enter senescence under proliferative pressure. Similarly, reduction of MMP-10 levels in young satellite cells from wild type animals induces a senescence response, while addition of the protease delays this program. Significantly, the effect of MMP-10 on satellite cell aging can be extended to another context of muscle wasting, muscular dystrophy. Systemic treatment of mdx dystrophic mice with MMP-10 prevents the muscle deterioration phenotype and reduces cellular damage in the satellite cells, which are normally under replicative pressure. Most importantly, MMP-10 conserves its protective effect in the satellite cell-derived myoblasts isolated from a Duchenne muscular dystrophy patient by decreasing the accumulation of damaged DNA. Hence, MMP-10 provides a previously unrecognized therapeutic opportunity to delay satellite cell aging and overcome satellite cell dysfunction in dystrophic muscles.

Deficiency of MMP-10 Aggravates the Diseased Phenotype of Aged Dystrophic Mice.

Matrix metalloproteinases (MMPs) have been implicated in the progression of muscular dystrophy, and recent studies have reported the role of MMP-10 in skeletal muscle pathology of young dystrophic mice. Nevertheless, its involvement in dystrophin-deficient hearts remains unexplored. Here, we aimed to investigate the involvement of MMP-10 in the progression of severe muscular dystrophy and to characterize MMP-10 loss in skeletal and cardiac muscles of aged dystrophic mice. We examined the histopathological effect of MMP-10 ablation in aged mdx mice, both in the hind limb muscles and heart tissues. We found that MMP-10 loss compromises survival rates of aged mdx mice, with skeletal and cardiac muscles developing a chronic inflammatory response. Our findings indicate that MMP-10 is implicated in severe muscular dystrophy progression, thus identifying a new area of research that could lead to future therapies for dystrophic muscles.

Association of SDF1 and MMP12 with Atherosclerosis and Inflammation: Clinical and Experimental Study.

BACKGROUND: Atherosclerosis is the main etiology of cardiovascular diseases (CVD), associated to systemic inflammation. Matrix metalloproteinases (MMPs) are related to atherosclerosis progression through the SDF1/CXCR4 axis promoting macrophages recruitment within the vascular wall. The goal was to assess new circulatory inflammatory markers in relation to atherosclerosis. METHODS: Measurement of SDF1, MMP12 and CRP in blood samples of 298 prospective patients with cardiovascular risk. To explore atherosclerosis progression, CXCR4/SDF1 axis and MMP12 expression were determined by RT-qPCR and by immunohistochemistry in the aorta of accelerated and delayed atherosclerosis mice models (Apoe-/- and Apoe-/-Mmp10-/-). RESULTS: SDF1, MMP12 and CRP were elevated in patients with clinical atherosclerosis, but after controlling by confounding factors, only SDF1 and CRP remained increased. Having high levels of both biomarkers showed 2.8-fold increased risk of presenting clinical atherosclerosis (p = 0.022). Patients with elevated SDF1, MMP12 and CRP showed increased risk of death in follow-up (HR = 3.2, 95%CI: 1.5-7.0, p = 0.004). Gene and protein expression of CXCR4 and MMP12 were increased in aortas from Apoe-/- mice. CONCLUSIONS: The combination of high circulating SDF1, MMP12 and CRP identified patients with particular inflammatory cardiovascular risk and increased mortality. SDF1/CXCR4 axis and MMP12 involvement in atherosclerosis development suggests that they could be possible atherosclerotic targets.

Reply to "Comment on 'Quantum fidelity approach to the ground-state properties of the one-dimensional axial next-nearest-neighbor Ising model in a transverse field' ".

We disagree with the objections raised by Nemati et al. regarding the phase transitions reported in our paper, where we used the fidelity method. Contrary to their claims, our fidelity calculations do not depend on energy level crossing between excited states. We obtain the same results just by analyzing the second derivative of the ground-state energy with respect to the interaction energy coupling J_{2}.

Ground-state properties of the one-dimensional transverse Ising model in a longitudinal magnetic field.

The critical properties of the one-dimensional spin-1/2 transverse Ising model in the presence of a longitudinal magnetic field were studied by the quantum fidelity method. We used exact diagonalization to obtain the ground-state energies and corresponding eigenvectors for lattice sizes up to 24 spins. The maximum of the fidelity susceptibility was used to locate the various phase boundaries present in the system. The type of dominant spin ordering for each phase was identified by examining the corresponding ground-state eigenvector. For a given antiferromagnetic nearest-neighbor interaction J_{2}, we calculated the fidelity susceptibility as a function of the transverse field B_{x} and the longitudinal field B_{z}. The phase diagram in the (B_{x},B_{z})-plane shows three phases. These findings are in contrast with the published literature that claims that the system has only two phases. For B_{x}<1, we observed an antiferromagnetic phase for small values of B_{z} and a paramagnetic phase for large values of B_{z}. For B_{x}>1 and low B_{z}, we found a disordered phase that undergoes a second-order phase transition to a paramagnetic phase for large values of B_{z}.

Quantum fidelity approach to the ground-state properties of the one-dimensional axial next-nearest-neighbor Ising model in a transverse field.

In this work we analyze the ground-state properties of the s=1/2 one-dimensional axial next-nearest-neighbor Ising model in a transverse field using the quantum fidelity approach. We numerically determined the fidelity susceptibility as a function of the transverse field B_{x} and the strength of the next-nearest-neighbor interaction J_{2}, for systems of up to 24 spins. We also examine the ground-state vector with respect to the spatial ordering of the spins. The ground-state phase diagram shows ferromagnetic, floating, and 〈2,2〉 phases, and we predict an infinite number of modulated phases in the thermodynamic limit (L→∞). Paramagnetism only occurs for larger magnetic fields. The transition lines separating the modulated phases seem to be of second order, whereas the line between the floating and the 〈2,2〉 phases is possibly of first order.

Dynamics of the transverse Ising model with next-nearest-neighbor interactions.

We study the effects of next-nearest-neighbor (NNN) interactions on the dynamics of the one-dimensional spin-1/2 transverse Ising model in the high-temperature limit. We use exact diagonalization to obtain the time-dependent transverse correlation function and the corresponding spectral density for a tagged spin. Our results for chains of 13 spins with periodic boundary conditions produce results which are valid in the infinite-size limit. In general we find that the NNN coupling produces slower dynamics accompanied by an enhancement of the central mode behavior. Even in the case of a strong transverse field, if the NNN coupling is sufficiently large, then there is a crossover from collective mode to central mode behavior. We also obtain several recurrants for the continued fraction representation of the relaxation function.

Quantum phase transitions in a chain with two- and four-spin interactions in a transverse field.

We use entanglement entropy and finite-size scaling methods to investigate the ground-state properties of a spin-1/2 Ising chain with two-spin (J(2)) and four-spin (J(4)) interactions in a transverse magnetic field (B). We concentrate our study on the unexplored critical region B=1 and obtain the phase diagram of the model in the (J(4)-J(2)) plane. The phases found include ferromagnetic (F), antiferromagnetic (AF), as well as more complex phases involving spin configurations with multiple periodicity. The system presents both first- and second-order transitions separated by tricritical points. We find an unusual phase boundary on the semi-infinite segment (J(4)<-1,J(2)=0) separating the F and AF phases.

Critical behavior of a quantum chain with four-spin interactions in the presence of longitudinal and transverse magnetic fields.

We study the ground-state properties of a spin-1/2 model on a chain containing four-spin Ising-like interactions in the presence of both transverse and longitudinal magnetic fields. We use entanglement entropy and finite-size scaling methods to obtain the phase diagrams of the model. Our numerical calculations reveal a rich variety of phases and the existence of multicritical points in the system. We identify phases with both ferromagnetic and antiferromagnetic orderings. We also find periodically modulated orderings formed by a cluster of like spins followed by another cluster of opposite like spins. The quantum phases in the model are found to be separated by either first- or second-order transition lines.

Phase transitions in the two-dimensional superantiferromagnetic Ising model with next-nearest-neighbor interactions.

We use Monte Carlo and transfer matrix methods in combination with extrapolation schemes to determine the phase diagram of the two-dimensional superantiferromagnetic (SAF) Ising model with next-nearest-neighbor (NNN) interactions in a magnetic field. The interactions between nearest-neighbor (NN) spins are ferromagnetic along x, and antiferromagnetic along Y. We find that for sufficiently low temperatures and fields, there exists a region limited by a critical line of second-order transitions separating a SAF phase from a magnetically induced paramagnetic phase. We did not find any region with either first-order transition or with reentrant behavior. The NNN couplings produce either an expansion or a contraction of the SAF phase. Expansion occurs when the interactions are antiferromagnetic, and contraction when they are ferromagnetic. There is a critical ratio R(c)=1/2 between NNN and NN couplings, beyond which the SAF phase no longer exists.

Quantum chaotic trajectories in integrable right triangular billiards.

Right triangular billiards are very simple systems that are completely integrable in classical mechanics for acute angle pairs (45 degrees,45 degrees ) and (30 degrees,60 degrees ). In quantum mechanics, the energy level spacing distribution of these billiards are neither Poisson-like nor Wigner-like. We use Bohm's formalism to calculate the trajectories, by numerical methods, for a particle inside these billiards. We use a linear combination of the first three energy states as the initial wave function. We show that a particle can have quasiperiodic or chaotic behavior, depending on its initial position in the billiards.

Lactobacillus plantarum amylase acting on crude starch granules. Native isoforms and activity changes after limited proteolysis.

The microheterogeneous native amylolytic complex secreted by the isolate A6 of Lactobacillus plantarum revealed a selective enzyme specificity loss when submitted to a limited proteolysis under a suboptimum pH condition. A clear electrophoretic profile change toward just one shorter, more acidic, and equally active polypeptide fragment resulted from the pronase E pretreatment. Although the whole enzyme activity remained apparently unaffected for soluble starch, the native parallel activity on intact and non-gelatinized starch granules either from cereals or tubers was dramatically reduced. This phenomenon was more clearly documented by scanning electron microscopy using the easiest accessible native substrate: wheat starch granules. The anion-exchange-purified native enzymes from L. plantarum displayed a different optimum pH curve when compared with the thermotolerant alpha-amylase from Bacillus licheniformis. The alpha-amylases from the lactic-acid-producing A6 isolate presented an electrophoretic profile easily distinguishable from those from B. liqueniformis and B. subtilis species.

Astaxanthinogenesis in the yeast Phaffia rhodozyma : optimization of low-cost culture media and yeast cell-wall lysis.

Astaxanthin is a diketo-dihydroxy-carotenoid produced by Phaffia rhodozyma, a basidiomicetous yeast. A low-cost fermentation medium consisting of raw sugarcane juice and urea was developed to exploit the active sucrolytic/urelolytic enzyme apparatus inherent to the yeast. As compared to the beneficial effect of 0.1 g% urea, a ready nitrogen source, mild phosphoric pre inversion of juice sucrose to glucose and fructose, promptly fermentable carbon sources, resulted in smaller benefits. Corn steep liquor (CSL) was found to be a valuable supplement for both yeast biomass yield (9.2 g dry cells/L) and astaxanthin production (1.3 mg/g cells). Distillery effluent (vinace), despite only a slightly positive effect on yeast growth, allowed for the highest pigment productivity (1.9 mg/g cells). Trace amounts of Ni2 (1 mg/L, as a cofactor for urease) resulted in controversial effects, namely, biomass decrease and astaxanthin increase, with no effect on the release (and uptake) of ammonium ion from urea. Since the synthesized astaxanthin is associated with the yeast cell and the pigment requires facilitated release for aquaculture uses (farmed fish meat staining), an investigation of the yeast cell wall was undertaken using detergent-treated cells. The composition of the rigid yeast envelope was found to be heterogeneous. Its partial acid or enzymatic depolymerization revealed glucose and xylose as common monomeric units of the cell-wall glycopolymers. Yeast cell-wall partial depolymerization with appropriate hydrolases may improve the pigment bioavailability for captive aquatic species and poultry.