A MicroRNA Perspective on Cardiovascular Development and Diseases: An Update.

In this review, we summarize the latest research pertaining to MicroRNAs (miRs) related to cardiovascular diseases. In today's molecular age, the key clinical aspects of diagnosing and treating these type of diseases are crucial, and miRs play an important role. Therefore, we have made a thorough analysis discussing the most important candidate protagonists of many pathways relating to such conditions as atherosclerosis, heart failure, myocardial infarction, and congenital heart disorders. We approach miRs initially from the fundamental molecular aspects and look at their role in developmental pathways, as well as regulatory mechanisms dysregulated under specific cardiovascular conditions. By doing so, we can better understand their functional roles. Next, we look at therapeutic aspects, including delivery and inhibition techniques. We conclude that a personal approach for treatment is paramount, and so understanding miRs is strategic for cardiovascular health.

Internal ribosome entry site (IRES) from Encephalomyocarditis virus (EMCV) as a tool for shuttle expression plasmids.

In eukaryotes, IRES sequences aid the recruitment of factors needed for translation to occur, enabling protein production independent of 5' capped mRNA. Many patents and commercially available plasmids exploit their properties for polycistronic expression of recombinant proteins. However, these applications have been restricted to eukaryotic organisms, since it was thought that elements of this origin were essential for their activity. Here, using two tricistronic vectors designed for expression in mammalian hosts, we present evidence of EMCV IRES activity in prokaryotes. This finding enables the development of new and more versatile plasmid vectors for the production of recombinant proteins in multiple hosts from a single construct. Additionally, it provides new hints for the elaboration of alternative models describing the molecular mechanism of EMCV IRES mediated translation, in the absence of eukaryotic elements that were considered indispensable for its function.

Diagnostic accuracy of buffy coat culture compared to total blood culture in late-onset sepsis of the newborn.

OBJECTIVES: To study the potential of buffy coat culture as a diagnostic tool for neonatal late-onset sepsis. METHODS: This was a study of diagnostic accuracy in newborn infants born at 28-41 weeks of gestation, weighing >800g, with ≥8 points on the NOSEP-1 scale. Paired samples for total blood culture (TBC) and buffy coat culture were drawn. We established the positivity rate, sensitivity, specificity, predictive values, and likelihood ratios, and compared time to positivity and contamination rates. RESULTS: Fifty-two newborns were included in the study. Twenty-one TBC and 22 buffy coat cultures were positive. The positivity rate for TBC was 40.4% and for buffy coat culture was 42.3% (p=not significant). Three TBC were positive with negative buffy coat culture. Four buffy coat cultures were positive with negative TBC; Kappa agreement was 0.723, p <0.001. Buffy coat culture sensitivity was 86% (95% confidence interval (CI) 68.5-95.4%), specificity 87% (75.4-93.7%), positive predictive value 82% (65.4-91.1%), negative predictive value 90% (77.9-96.8%), positive likelihood ratio 6.64 (2.79-15.05), and negative likelihood ratio 0.16 (0.05-0.42). We found no difference in time to positivity in hours; Wilcoxon Z=1224, p=0.22. The contamination rate was 1.9% for both methods. CONCLUSIONS: Buffy coat culture is as good as TBC for the microbiological diagnosis of late-onset sepsis of the newborn. Buffy coat culture allows the use of remaining plasma for further analysis.