These data suggest that furthermore for the direct protein regulation, microRNAs may possibly also modulate qualitative traits of chosen proteins by an indirect handle of post-translational processes. Search phrases: miRNAs, Sialytransferases, Antithrombin, Post-translational modifications, Microarray, Post-natal developmentBackground MicroRNAs (miRNAs) are modest non-coding RNAs implicated within the modulation of a large number of physiological and pathological processes [1,2] by way of a mechanism primarily based on the repression of protein translation or degradation of messenger RNAs [3]. MiRNAs happen to be lately involved inside the modulation of various haemostatic factors such as fibrinogen, tissue factor, and proteins implicated in platelet function [4-6]. Actually, miRNAs can also be involved in the quantitative variations of components on the haemostatic system observed through development [7]. In humans, levels of haemostatic aspects go in constant raise soon after birth until reaching levels related to these located in adults within the first year [8-10]. In specific, antithrombin, an anticoagulant serpin crucial in the manage of your hemostatic method [11], is substantially lowered (50 ) in plasma of neonates in comparison with adults* Correspondence: [email protected] Centro Regional de Hemodonaci , University of Murcia, IMIB, Spain, C/Ronda de Garay S/N, 30003, Murcia, Spain[8]. Despite these variations, neonates sustain a perfect haemostatic equilibrium. Accordingly, a developmental model involving neonatal and adult period is ideal to study the mechanisms that regulate haemostatic protein levels and also the adaptation of this technique to distinct circumstances. Additionally to these quantitative alterations, few functions have shown that neonatal antithrombin has reduced levels of sialic acid than its adult counterpart but the molecular mechanism of this regulation is unknown [12,13]. Hence, the developmental model may perhaps also let to investigating the regulation of N-glycosylation by miRNAs and hence to further enlarge the effects of miRNAs in gene regulation.MethodsMouse samplesNon-inbred Swiss CD1 mice from distinctive litters had been sacrificed by cervical dislocation or decapitation at various ages, from day one particular right after birth to adult age.Salvianolic acid A MMP Livers finely dissected had been right away snap-frozen in liquid nitrogen.Tetrapropylammonium perruthenate web Blood was anticoagulated with trisodium citrate,2013 Teruel et al.PMID:35345980 ; licensee BioMed Central Ltd. This is an Open Access write-up distributed below the terms from the Inventive Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, offered the original operate is appropriately cited.Teruel et al. Journal of Biomedical Science 2013, 20:29 http://www.jbiomedsci/content/20/1/Page 2 ofcentrifuged at 1,500 g for 5 minutes to receive platelet poor plasma and promptly stored in aliquots at -80 . All experimental procedures strictly followed the University of Murcia authorized Institutional Animal Care guidelines and had been approved by the local ethical committee (#C131002043; 15/02/2010).Antithrombin levels and activityMicroRNA microarrayAntithrombin activity was determined by chromogenic methods, as previously described [14]. Anti-factor Xa (antiFXa) assay was performed with pentasaccharide, bovine FXa, and S-2765 chromogenic substrate (Chromogenix, IZASA, Spain). Antithrombin levels were determined by enzymelinked immunosorbent assay and electro-immunodiffusion (Laurell), as previously reported [1.
