Rodrigo Jácome

Structural Analysis of Monomeric RNA-Dependent Polymerases: Evolutionary and Therapeutic Implications.

The crystal structures of monomeric RNA-dependent RNA polymerases and reverse transcriptases of more than 20 different viruses are available in the Protein Data Bank. They all share the characteristic right-hand shape of DNA- and RNA polymerases formed by the fingers, palm and thumb subdomains, and, in many cases, “fingertips” that extend from the fingers towards the thumb subdomain, giving the viral enzyme a closed right-hand appearance. Six conserved structural motifs that contain key residues for the proper functioning of the enzyme have been identified in all these RNA-dependent polymerases. These enzymes share a two divalent metal-ion mechanism of polymerization in which two conserved aspartate residues coordinate the interactions with the metal ions to catalyze the nucleotidyl transfer reaction. The recent availability of crystal structures of polymerases of the Orthomyxoviridae and Bunyaviridae families allowed us to make pairwise comparisons of the tertiary structures of polymerases belonging to the four main RNA viral groups, which has led to a phylogenetic tree in which single-stranded negative RNA viral polymerases have been included for the first time. This has also allowed us to use a homology-based structural prediction approach to develop a general three-dimensional model of the Ebola virus RNA-dependent RNA polymerase. Our model includes several of the conserved structural motifs and residues described in other viral RNA-dependent RNA polymerases that define the catalytic and highly conserved palm subdomain, as well as portions of the fingers and thumb subdomains. The results presented here help to understand the current use and apparent success of antivirals, i.e. Brincidofovir, Lamivudine and Favipiravir, originally aimed at other types of polymerases, to counteract the Ebola virus infection.

Endothelial Dysfunction in Systemic Lupus Erythematosus: Evaluation with 13N-Ammonia PET

Systemic lupus erythematosus (SLE) affects multiple organs and systems, severely involving the cardiovascular system. The aim of this study was to evaluate the presence of endothelial dysfunction with 13N-ammonia PET in asymptomatic SLE patients. Methods: We enrolled 16 women with SLE and 16 healthy women. Myocardial blood flow (MBF) was quantified in a 64-slice PET/CT scanner at rest, during a cold pressor test (CPT), and during stress. Endothelium-dependent vasodilation index, %ΔMBF, and myocardial flow reserve (MFR) were calculated. Results: There were 16 women in the SLE group (mean age ± SD, 31.4 ± 8.3 y) and 16 women in the healthy control group (31.5 ± 11.1 y). Mean endothelium-dependent vasodilatation index and %ΔMBF were significantly lower in SLE patients (1.18 ± 0.55 vs. 1.63 ± 0.65, P = 0.04, and 18 ± 55 vs. 63 ± 65, P = 0.04, respectively). MFR was also lower in the SLE group (2.41 ± 0.59 vs. 2.73 ± 0.77, P = 0.20). Conclusion: SLE patients who are free of active disease present abnormal coronary flow and endothelial dysfunction. It is necessary to develop and intensify treatment strategies directed to CAD in SLE patients.

Coenzymes, viruses and the RNA world.

The results of a detailed bioinformatic search for ribonucleotidyl coenzyme biosynthetic sequences in DNA- and RNA viral genomes are presented. No RNA viral genome sequence available as of April 2011 appears to encode for sequences involved in coenzyme biosynthesis. In both single- and double-stranded DNA viruses a diverse array of coenzyme biosynthetic genes has been identified, but none of the viral genomes examined here encodes for a complete pathway. Although our conclusions may be constrained by the unexplored diversity of viral genomes and the biases in the construction of viral genome databases, our results do not support the possibility that RNA viruses are direct holdovers from an ancient RNA/protein world. Extrapolation of our results to evolutionary epochs prior to the emergence of DNA genomes suggest that during those early stages living entities may have depended on discontinuous genetic systems consisting of multiple small-size RNA sequences.

Functional Impact of Coronary Stenosis Observed on Coronary Computed Tomography Angiography: Comparison with 13N-Ammonia PET

BACKGROUND AND AIMS We undertook this study to evaluate the functional impact of coronary abnormalities in patients with suspected coronary artery disease (CAD) by means of integrated positron emission tomography (PET) and coronary computed tomography angiography (CCTA) scan obtained on a hybrid state-of-the-art PET/CT scanner. METHODS We studied 29 consecutive, patients with a clinically suspected intermediate risk for CAD, using a hybrid PET/CT 64 slice scanner. During a single scanning session, CCTA was performed for coronary anatomy evaluation, and a rest/adenosine stress (13)N-ammonia PET was performed for myocardial perfusion assessment in 3D mode with CT attenuation correction. RESULTS Twenty four (82.7%) patients had atherosclerosis detected by CCTA; 15 patients had significant (≥50%) coronary stenoses and all 15 patients showed ischemia by PET; moreover, 10/15 patients had a Summed Stress Score >12.20/24 and 83.3% patients with atherosclerosis detected by CCTA showed ischemia by PET. Two of five patients with normal coronary arteries showed ischemia by PET. CCTA agreement in positive identification of PET ischemia was 91% and agreement in ruling out ischemia was 43%; PET agreement in detecting CCTA atherosclerosis was 83%, and agreement in ruling it out was 60%. CONCLUSIONS We found a strong relation between significant coronary stenosis identified by CCTA and ischemia by PET. However, in cases with low-grade stenosis, PET scan can assess the functional significance of atherosclerotic abnormalities.

Are RNA Viruses Candidate Agents for the Next Global Pandemic? A Review

Abstract Pathogenic RNA viruses are potentially the most important group involved in zoonotic disease transmission, and they represent a challenge for global disease control. Their biological diversity and rapid adaptive rates have proved to be difficult to overcome and to anticipate by modern medical technology. Also, the anthropogenic change of natural ecosystems and the continuous population growth are driving increased rates of interspecies contacts and the interchange of pathogens that can develop into global pandemics. The combination of molecular, epidemiological, and ecological knowledge of RNA viruses is therefore essential towards the proper control of these emergent pathogens. This review outlines, throughout different levels of complexity, the problems posed by RNA viral diseases, covering some of the molecular mechanisms allowing them to adapt to new host species—and to novel pharmaceutical developments—up to the known ecological processes involved in zoonotic transmission.