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"I AM A WOMAN OF GOD" WOMAN'S CONFERENCE Group

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Emmanuel Konovalov
Emmanuel Konovalov

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We measured the levels of miR-574-5p, a microRNA involved in neuronal death, in the serum of patients who suffered cardiac arrest and were resuscitated. We compared the levels of miR-574-5p between patients who had a good neurological outcome (CPC 1-2) and those who had a poor neurological outcome (CPC 3-5) at 72 hours after cardiac arrest. We found that patients with poor neurological outcome had significantly higher levels of miR-574-5p than patients with good neurological outcome (Figure 2(a)). This difference was not affected by the sex of the patients (Figures 2(b) and 2(c)), or by the targeted temperature management regimen they received (33ÂC vs. 36ÂC, Supplementary Figure 2a-f). However, we observed that patients who were treated at 33ÂC had higher levels of miR-574-5p than patients who were treated at 36ÂC (Supplementary Figure 2g), regardless of their sex (Supplementary Figure 2h-i). This suggests that miR-574-5p may be influenced by the cooling therapy and may have a role in the neuroprotective effects of hypothermia.


To further investigate the role of miR-574-5p in neuronal injury after cardiac arrest, we performed in vitro experiments using primary cultures of rat cortical neurons. We exposed the neurons to oxygen-glucose deprivation (OGD), a model of ischemia-reperfusion injury, and measured the levels of miR-574-5p and its target genes. We found that OGD induced a significant increase in miR-574-5p expression and a decrease in the expression of its target genes, which are involved in neuronal survival and plasticity (Figure 3(a) and 3(b)). We also found that overexpression of miR-574-5p by transfection of a miR-574-5p mimic enhanced OGD-induced neuronal death, while inhibition of miR-574-5p by transfection of a miR-574-5p inhibitor attenuated OGD-induced neuronal death (Figure 3(c) and 3(d)). These results indicate that miR-574-5p is a mediator of neuronal death after ischemia-reperfusion injury.


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To evaluate the effect of hypothermia on miR-574-5p expression and neuronal survival, we exposed the neurons to OGD at either 37ÂC or 33ÂC. We found that hypothermia reduced the levels of miR-574-5p and increased the levels of its target genes after OGD (Figure 4(a) and 4(b)). Moreover, hypothermia prevented OGD-induced neuronal death, as shown by reduced lactate dehydrogenase (LDH) release and increased cell viability (Figure 4(c) and 4(d)). These effects were reversed by overexpression of miR-574-5p, suggesting that hypothermia exerts its neuroprotective effects by downregulating miR-574-5p.


In order to confirm the involvement of miR-574-5p in the neuroprotective effects of hypothermia in vivo, we performed animal experiments using a rat model of cardiac arrest and resuscitation. We randomly assigned the rats to four groups: normothermia (37ÂC) with saline injection, normothermia with miR-574-5p inhibitor injection, hypothermia (33ÂC) with saline injection, and hypothermia with miR-574-5p mimic injection. We induced cardiac arrest by asphyxia and resuscitated the rats by chest compression and ventilation. We then maintained the rats at either 37ÂC or 33ÂC for 6 hours and injected either saline, miR-574-5p inhibitor, or miR-574-5p mimic into the lateral ventricle. We measured the levels of miR-574-5p and its target genes in the brain tissue at 24 hours after resuscitation. We also assessed the neurological outcome of the rats at 72 hours after resuscitation using a neurological deficit score (NDS).


We found that cardiac arrest and resuscitation induced a significant increase in miR-574-5p levels and a decrease in its target gene levels in the brain tissue (Figure 5(a) and 5(b)). Hypothermia reduced the levels of miR-574-5p and increased the levels of its target genes compared to normothermia (Figure 5(a) and 5(b)). Injection of miR-574-5p inhibitor further reduced the levels of miR-574-5p and increased the levels of its target genes in both normothermic and hypothermic rats (Figure 5(a) and 5(b)). Injection of miR-574-5p mimic reversed the effects of hypothermia on miR-574-5p and its target genes (Figure 5(a) and 5(b)). These results indicate that hypothermia modulates miR-574-5p expression in the brain after cardiac arrest and resuscitation. 0efd9a6b88


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