The hypoxia sensitive metal transcription factor MTF-1 activates NCX1 brain promoter and participates in remote postconditioning neuroprotection in stroke

Remote limb ischemic postconditioning (RLIP) is an experimental strategy in which short femoral artery ischemia reduces brain damage induced by a previous harmful ischemic insult. Ionic homeostasis maintenance in the CNS seems to play a relevant role in mediating RLIP neuroprotection and among the effectors, the sodium-calcium exchanger 1 (NCX1) may give an important contribution, being expressed in all CNS cells involved in brain ischemic pathophysiology. The aim of this work was to investigate whether the metal responsive transcription factor 1 (MTF-1), an important hypoxia sensitive transcription factor, may (i) interact and regulate NCX1, and (ii) play a role in the neuroprotective effect mediated by RLIP through NCX1 activation. Here we demonstrated that in brain ischemia induced by transient middle cerebral occlusion (tMCAO), MTF-1 is triggered by a subsequent temporary femoral artery occlusion (FAO) and represents a mediator of endogenous neuroprotection. More importantly, we showed that MTF-1 translocates to the nucleus where it binds the metal responsive element (MRE) located at −23/−17 bp of Ncx1 brain promoter thus activating its transcription and inducing an upregulation of NCX1 that has been demonstrated to be neuroprotective. Furthermore, RLIP restored MTF-1 and NCX1 protein levels in the ischemic rat brain cortex and the silencing of MTF-1 prevented the increase of NCX1 observed in RLIP protected rats, thus demonstrating a direct regulation of NCX1 by MTF-1 in the ischemic cortex of rat exposed to tMCAO followed by FAO. Moreover, silencing of MTF-1 significantly reduced the neuroprotective effect elicited by RLIP as demonstrated by the enlargement of brain infarct volume observed in rats subjected to RLIP and treated with MTF-1 silencing. Overall, MTF-dependent activation of NCX1 and their upregulation elicited by RLIP, besides unraveling a new molecular pathway of neuroprotection during brain ischemia, might represent an additional mechanism to intervene in stroke pathophysiology

The evolution of the orbit distance in the double averaged restricted 3-body problem with crossing singularities

We study the long term evolution of the distance between two Keplerian confocal trajectories in the framework of the averaged restricted 3-body problem. The bodies may represent the Sun, a solar system planet and an asteroid. The secular evolution of the orbital elements of the asteroid is computed by averaging the equations of motion over the mean anomalies of the asteroid and the planet. When an orbit crossing with the planet occurs the averaged equations become singular. However, it is possible to define piecewise differentiable solutions by extending the averaged vector field beyond the singularity from both sides of the orbit crossing set. In this paper we improve the previous results, concerning in particular the singularity extraction technique, and show that the extended vector fields are Lipschitz-continuous. Moreover, we consider the distance between the Keplerian trajectories of the small body and of the planet. Apart from exceptional cases, we can select a sign for this distance so that it becomes an analytic map of the orbital elements near to crossing configurations. We prove that the evolution of the 'signed' distance along the averaged vector field is more regular than that of the elements in a neighborhood of crossing times. A comparison between averaged and non-averaged evolutions and an application of these results are shown using orbits of near-Earth asteroids.Comment: 29 pages, 8 figure

Simbol-X Hard X-ray Focusing Mirrors: Results Obtained During the Phase A Study

Simbol-X will push grazing incidence imaging up to 80 keV, providing a strong improvement both in sensitivity and angular resolution compared to all instruments that have operated so far above 10 keV. The superb hard X-ray imaging capability will be guaranteed by a mirror module of 100 electroformed Nickel shells with a multilayer reflecting coating. Here we will describe the technogical development and solutions adopted for the fabrication of the mirror module, that must guarantee an Half Energy Width (HEW) better than 20 arcsec from 0.5 up to 30 keV and a goal of 40 arcsec at 60 keV. During the phase A, terminated at the end of 2008, we have developed three engineering models with two, two and three shells, respectively. The most critical aspects in the development of the Simbol-X mirrors are i) the production of the 100 mandrels with very good surface quality within the timeline of the mission; ii) the replication of shells that must be very thin (a factor of 2 thinner than those of XMM-Newton) and still have very good image quality up to 80 keV; iii) the development of an integration process that allows us to integrate these very thin mirrors maintaining their intrinsic good image quality. The Phase A study has shown that we can fabricate the mandrels with the needed quality and that we have developed a valid integration process. The shells that we have produced so far have a quite good image quality, e.g. HEW <~30 arcsec at 30 keV, and effective area. However, we still need to make some improvements to reach the requirements. We will briefly present these results and discuss the possible improvements that we will investigate during phase B.Comment: 6 pages, 3 figures, invited talk at the conference "2nd International Simbol-X Symposium", Paris, 2-5 december, 200

sodium calcium exchanger as main effector of endogenous neuroprotection elicited by ischemic tolerance

Abstract The ischemic tolerance (IT) paradigm represents a fundamental cell response to certain types or injury able to render an organ more "tolerant" to a subsequent, stronger, insult. During the 16th century, the toxicologist Paracelsus described for the first time the possibility that a noxious event might determine a state of tolerance. This finding was summarized in one of his most important mentions: "The dose makes the poison". In more recent years, ischemic tolerance in the brain was first described in 1991, when it was demonstrated by Kirino and collaborators that two minutes of subthreshold brain ischemia in gerbils produced tolerance against global brain ischemia. Based on the time in which the conditioning stimulus is applied, it is possible to define preconditioning, perconditioning and postconditioning, when the subthreshold insult is applied before, during or after the ischemic event, respectively. Furthermore, depending on the temporal delay from the ischemic event, two different modalities are distinguished: rapid or delayed preconditioning and postconditioning. Finally, the circumstance in which the conditioning stimulus is applied on an organ distant from the brain is referred as remote conditioning. Over the years the "conditioning" paradigm has been applied to several brain disorders and a number of molecular mechanisms taking part to these protective processes have been described. The mechanisms are usually classified in three distinct categories identified as triggers, mediators and effectors. As concerns the putative effectors, it has been hypothesized that brain cells appear to have the ability to adapt to hypoxia by reducing their energy demand through modulation of ion channels and transporters, which delays anoxic depolarization. The purpose of the present review is to summarize the role played by plasmamembrane proteins able to control ionic homeostasis in mediating protection elicited by brain conditioning, particular attention will be deserved to the role played by Na+/Ca2+ exchanger

miR135a administration ameliorates brain ischemic damage by preventing TRPM7 activation during brain ischemia

Background: miRNA-based strategies have recently emerged as a promising therapeutic approach in several neurodegenerative diseases. Unregulated cation influx is implicated in several cellular mechanisms underlying neural cell death during ischemia. The brain constitutively active isoform of transient receptor potential melastatin 7 (TRPM7) represents a glutamate excitotoxicity-independent pathway that significantly contributes to the pathological Ca2+ overload during ischemia. Aims: In the light of these premises, inhibition of TRPM7 may be a reasonable strategy to reduce ischemic injury. Since TRPM7 is a putative target of miRNA135a, the aim of the present paper was to evaluate the role played by miRNA135a in cerebral ischemia. Therefore, the specific objectives of the present paper were: (1) to evaluate miR135a expression in temporoparietal cortex of ischemic rats; (2) to investigate the effect of the intracerebroventricular (icv) infusion of miR135a on ischemic damage and neurological functions; and (3) to verify whether miR135a effects may be mediated by an alteration of TRPM7 expression. Methods: miR135a expression was evaluated by RT- PCR and FISH assay in temporoparietal cortex of ischemic rats. Ischemic volume and neurological functions were determined in rats subjected to transient middle cerebral artery occlusion (tMCAo) after miR135a intracerebroventricular perfusion. Target analysis was performed by Western blot. Results: Our results demonstrated that, in brain cortex, 72 h after ischemia, miR135a expression increased, while TRPM7 expression was parallelly downregulated. Interestingly, miR135a icv perfusion strongly ameliorated the ischemic damage and improved neurological functions, and downregulated TRPM7 protein levels. Conclusions: The early prevention of TRPM7 activation is protective during brain ischemia

Prolonged NCX activation prevents SOD1 accumulation, reduces neuroinflammation, ameliorates motor behavior and prolongs survival in a ALS mouse model

Imbalance in cellular ionic homeostasis is a hallmark of several neurodegenerative diseases including Amyotrophic Lateral Sclerosis (ALS). Sodium-calcium exchanger (NCX) is a membrane antiporter that, operating in a bidirectional way, couples the exchange of Ca2+ and Na + ions in neurons and glial cells, thus controlling the intracellular homeostasis of these ions. Among the three NCX genes, NCX1 and NCX2 are widely expressed within the CNS, while NCX3 is present only in skeletal muscles and at lower levels of expression in selected brain regions. ALS mice showed a reduction in the expression and activity of NCX1 and NCX2 consistent with disease progression, therefore we aimed to investigate their role in ALS pathophysiology. Notably, we demonstrated that the pharmacological activation of NCX1 and NCX2 by the prolonged treatment of SOD1G93A mice with the newly synthesized compound neurounina: (1) prevented the reduction in NCX activity observed in spinal cord; (2) preserved motor neurons survival in the ventral spinal horn of SOD1G93A mice; (3) prevented the spinal cord accumulation of misfolded SOD1; (4) reduced astroglia and microglia activation and spared the resident microglia cells in the spinal cord; (5) improved the lifespan and mitigated motor symptoms of ALS mice. The present study highlights the significant role of NCX1 and NCX2 in the pathophysiology of this neurodegenerative disorder and paves the way for the design of a new pharmacological approach for ALS

In-stent restenosis in the drug eluting stent assayed by optical coherence tomography

Hospital of Holy Pope Giovanni XXIII, Bergamo, Italy, Department of Interventional Cardiology, Institute of Cardiology, Chisinau, the Republic of MoldovaBackground: In order to study the mechanisms and morphological aspects of the in-stent restenosis (ISR) have been evaluated restenosis lesions referred to drug eluting stent (DES) of the first generation using optical coherence tomography technique (OCT). Material and methods: The study underwent 39 patients with ISR induced recurrent angina or acute coronary syndrome including 66 stents from which were: 44 stents (28 patients) of the first generation of DES (19 – Cypher TM and 23 – Taxus®), and 22 stents (11 patients) of the second DES generation (9 Xience TM 2 Promus TM), 10 ZES (Resolute TM), and one stent Nobori TM. Has been made quantitative and morphological analysis of tissue pattern of ISR using the following OCT criteria: 1) morphologically homogenous neointima; 2)morphologically heterogeneous neointima; and 3) neoatherosclerosis. Results: It has been established that in the first generation of DES morphological homogenous pattern was present in both ISR developed after 1 year and later. However, the optical aspect with heterogeneous presentation had a prevalence decline in dynamics. The heterogeneous model had a higher prevalence in actual generation of DES in both incipient (< 1 year) and late presentation. Conclusions: The phenomenon of neoatherosclerosis has presented a significantly less frequency in the late restenosis of actual generation of DES. Our results suggest that restenosis phenomenon in actual generation of DES has a different morphological and evolution pattern in time in comparison with ISR of the first generation of DES

Intracoronary optical coherence tomography

The cardiovascular departments of the hospital of Holy Pope Giovanni XXIII in Bergamo, Italy and of the Institute of Cardiology in Chisinau, the Republic of Moldova have a fruitful history of collaboration in the field of interventional cardiology and intracoronary imaging in particular. We have recently expanded our collaboration by adding the method of Optical Coherence Tomography (OCT) into the armamentarium of imaging in the catheterization laboratory. OCT is an innovative, real time, tomographic imaging modality able to visualize tissues at microstructure level. It delivers the rays of near-infrared light through the wall of the coronary artery using small diameter optical fibres. The light that illuminates the vessel is absorbed and backscattered or reflected by the structures of the tissues with different degrees of density, thus creating an image with an axial resolution of 10-20 µm. This technology allows acquiring high definition images of long segments of coronaries for a few seconds. For the time being, OCT is mainly used in the researches, providing insights into the pathophysiology of the atherosclerotic plaque and the vascular response to stenting. It also has a potential for clinical application, such as pre-interventional evaluation of coronary arteries, procedures guidance and follow-up assessment of vascular healing after the stent implantation. A joined database has been created by the two institutions in an effort to study in vivo the morphology of the coronary arteries in different pathologies. This review is focused on the potential fields of application of OCT in different clinical and scientific institutions