Assessment of the Therapeutic Potential of Metallothionein-II Application in Focal Cerebral Ischemia In Vitro and In Vivo

Metallothionein-II (MT-II) is an ubiquitously expressed small-molecular-weight protein and highly induced in various species and tissues upon stress, inflammation, and ischemia. MT-deficiency exacerbates ischemic injury in rodent stroke models in vitro and in vivo. However, there is conflicting data on the potential neuroprotective effect of exogenously applied metallothionein. Thus, we applied MT-II in an in vitro stroke model and intraperitoneally (i.p.) in two in vivo standard models of transient middle cerebral artery occlusion (MCAO) (a ‘stringent’ one [60min MCAO/48h reperfusion] and a ‘mild’ one [30min MCAO/72h reperfusion]), as well as i.v. together with recombinant tissue plasminogen activator (rtPA) to evaluate if exogenous MT-II-application protects against ischemic stroke. Whereas MT-II did not protect against 60min MCAO, there was a significant reduction of direct and indirect infarct volumes and neurological deficit in the MT-II (i.p.) treated animals in the ‘mild’ model at 3d after MCAO. Furthermore, MT- II also improved survival of the mice after MCAO, suppressed TNF-α mRNA induction in ischemic brain tissue, and protected primary neuronal cells against oxygen-glucose-deprivation in vitro. Thus, exogenous application of MT-II protects against ischemic injury in vitro and in vivo. However, long- term studies with different species and larger sampling sizes are required before a clinical use can be envisaged

Membrane attack complex inhibitor CD59a protects against focal cerebral ischemia in mice

<p>Abstract</p> <p>Background</p> <p>The complement system is a crucial mediator of inflammation and cell lysis after cerebral ischemia. However, there is little information about the exact contribution of the membrane attack complex (MAC) and its inhibitor-protein CD59.</p> <p>Methods</p> <p>Transient focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) in young male and female CD59a knockout and wild-type mice. Two models of MCAO were applied: 60 min MCAO and 48 h reperfusion, as well as 30 min MCAO and 72 h reperfusion. CD59a knockout animals were compared to wild-type animals in terms of infarct size, edema, neurological deficit, and cell death.</p> <p>Results and Discussion</p> <p>CD59a-deficiency in male mice caused significantly increased infarct volumes and brain swelling when compared to wild-type mice at 72 h after 30 min-occlusion time, whereas no significant difference was observed after 1 h-MCAO. Moreover, CD59a-deficient mice had impaired neurological function when compared to wild-type mice after 30 min MCAO.</p> <p>Conclusion</p> <p>We conclude that CD59a protects against ischemic brain damage, but depending on the gender and the stroke model used.</p

Serum calcification propensity is independently associated with disease activity in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is associated with severe cardiovascular complications. The T50 score is a novel functional blood test quantifying calcification propensity in serum. High calcification propensity (or low T50) is a strong and independent determinant of all-cause mortality in various patient populations. A total of 168 patients with ≥ 4 American College of Rheumatology (ACR) diagnostic criteria from the Swiss Systemic lupus erythematosus Cohort Study (SSCS) were included in this analysis. Serum calcification propensity was assessed using time-resolved nephelometry. The cohort mainly consisted of female (85%), middle-aged (43±14 years) Caucasians (77%). The major determinants of T50 levels included hemoglobin, serum creatinine and serum protein levels explaining 43% of the variation at baseline. Integrating disease activity (SELENA-SLEDAI) into this multivariate model revealed a significant association between disease activity and T50 levels. In a subgroup analysis considering only patients with active disease (SELENA-SLEDAI score ≥4) we found a negative association between T50 and SELENA-SLEDAI score at baseline (Spearman's rho -0.233, P = 0.02). Disease activity and T50 are closely associated. Moreover, T50 levels identify a subgroup of SLE patients with ongoing systemic inflammation as mirrored by increased disease activity. T50 could be a promising biomarker reflecting SLE disease activity and might offer an earlier detection tool for high-risk patients

The role of myostatin in muscle wasting: an overview

Myostatin is an extracellular cytokine mostly expressed in skeletal muscles and known to play a crucial role in the negative regulation of muscle mass. Upon the binding to activin type IIB receptor, myostatin can initiate several different signalling cascades resulting in the upregulation of the atrogenes and downregulation of the important for myogenesis genes. Muscle size is regulated via a complex interplay of myostatin signalling with the insulin-like growth factor 1/phosphatidylinositol 3-kinase/Akt pathway responsible for increase in protein synthesis in muscle. Therefore, the regulation of muscle weight is a process in which myostatin plays a central role but the mechanism of its action and signalling cascades are not fully understood. Myostatin upregulation was observed in the pathogenesis of muscle wasting during cachexia associated with different diseases (i.e. cancer, heart failure, HIV). Characterisation of myostatin signalling is therefore a perspective direction in the treatment development for cachexia. The current review covers the present knowledge about myostatin signalling pathways leading to muscle wasting and the state of therapy approaches via the regulation of myostatin and/or its downstream targets in cachexia

Effect of metallothionein-II <i>i</i>.<i>p</i>. treatment <i>in vivo</i> 48h after MCAO for 60 min.

<p>Infarct volumes of vehicle (0.9% NaCl) (n = 7) or MT-II (<i>i</i>.<i>p</i>.) treated (n = 8) male adult wildtype C57BL/6N mice after 60min MCAO and 48h reperfusion. <b>A</b>: direct infarct volumes. <b>B</b>: inflammatory cell count in the ischemic hemisphere as determined by counting Iba1-positive cells in whole ischemic brain hemispheres of mice at 48h of reperfusion after 60min MCAO. Inflammatory cell accumulation (macrophages and activated microglia) was determined by counting Iba1-positive cells at interaural position No.III (distance to bregma 3.9mm) in the whole ischemic/ipsilateral hemisphere. <b>C</b>: Brain swelling, as calculated by the difference between direct and indirect infarct volumes. Whereas indirect infarct volumes were calculated as the volume of the contralateral hemisphere minus the non-infarcted volume of the ipsilateral/ischemic hemisphere (p-values between groups were >0.05 as calculated by Mann Whitney U-Test, one-tailed). <b>D:</b> neurological deficit of mice at 48h after induction of MCAO as determined by a modified Bederson score [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144035#pone.0144035.ref041" target="_blank">41</a>]: 0 represents no deficits, 1 represents an extension deficit in the contralateral leg, 2 a hemiparesis with circling, 3 loss of postural reflexes, and 4 death.</p