Biophysical and atomic force microscopy characterization of the RNA from satellite tobacco mosaic virus

Agarose gel electrophoresis, circular dichroism and differential scanning calorimetry showed that single-stranded RNA from satellite tobacco mosaic virus transforms from a conformationally ‘closed state’ at 4°C to a more conformationally ‘open state’ at 65°C. The transition is reversible and shows no hysteresis. Atomic force microscopy (AFM) allowed visualization of the two states and indicated that the conformationally ‘closed state’ probably corresponds to the native encapsidated conformation, and that the ‘open state’ represents a conformation, characterized as short, thick chains of domains, as a consequence of the loss of tertiary interactions. Heating from 75°C to 85°C in the presence of EDTA was necessary to further unravel the ‘open’ conformation RNA into extended chains of lengths >280 nm. Virus exposed to low concentrations of phenol at 65°C, extruded RNA as distinctive ‘pigtails’ in a synchronous fashion, and these ‘pigtails’ then elongated, as the RNA was further discharged by the particles. Moderate concentrations of phenol at 65°C produced complete disruption of virions and only remains of decomposed particles and disordered RNA were evident. AFM images of RNA emerging from disrupted virions appear most consistent with linear arrangements of structural domains

Cardiac resynchronization therapy and valvular cardiomyopathy after corrective surgery

Cardiac resynchronization therapy (CRT) has been shown to have clinical benefits in certain groups of patients with advanced heart failure (HF). However, patients with valvular cardiomyopathy are underrepresented in randomized clinical studies. The aim of this study was to assess the medium-term (i.e., at 6 months) effects of CRT in patients with HF exclusively due to valvular disease. The study included 40 consecutive patients who underwent CRT device implantation. At 6 months, there were improvements in functional class, left ventricular remodeling, and intraventricular dyssynchrony parameters in treated patients. In this particular subgroup of patients, the benefits of CRT were similar to those observed in patients with HF due to other etiologies

Protein crystallization in short-peptide supramolecular hydrogels: A versatile strategy towards biotechnological composite materials

Protein crystallization in hydrogels has been explored with the main purpose of facilitating the growth of high quality crystals while increasing their size to enhance their manipulation. New avenues are currently being built for the use of protein crystals as source materials to create sensors and drug delivery vehicles, to name just a few. In this sense, short-peptide supramolecular hydrogels may play a crucial role in integrating protein crystals within a wider range of applications. In this article, we show that protein crystallization in short-peptide supramolecular hydrogels is feasible and independent of the type of peptide that forms the hydrogel and/or the protein, although the output is not always the same. As a general trend, it is confirmed that hydrogel fibers are always incorporated within crystals so that novel composite materials for biotechnological applications with enhanced properties are produced.This research was funded by the MICINN (Spain) projects BIO2010-6800 (JAG), CTQ2012-34778 (JJDM), and “Factoría Española de Cristalización” Consolider-Ingenio 2010 (JAG & MCM), and by Junta de Andalucía (Spain) project P12-FQM- 2721 (LAC). EDRF funds JAG, LAC & JMC. JJDM thanks MICINN for a Ramon y Cajal Fellowship and MCM thanks CSIC for her JAE Fellowshi

Efficacy of aldose reductase inhibitors is affected by oxidative stress induced under X-ray irradiation

Human aldose reductase (hAR, AKR1B1) has been explored as drug target since the 1980s for its implication in diabetic complications. An activated form of hAR was found in cells from diabetic patients, showing a reduced sensitivity to inhibitors in clinical trials, which may prevent its pharmacological use. Here we report the conversion of native hAR to its activated form by X-ray irradiation simulating oxidative stress conditions. Upon irradiation, the enzyme activity increases moderately and the potency of several hAR inhibitors decay before global protein radiation damage appears. The catalytic behavior of activated hAR is also reproduced as the KM increases dramatically while the kcat is not much affected. Consistently, the catalytic tetrad is not showing any modification. The only catalytically-relevant structural difference observed is the conversion of residue Cys298 to serine and alanine. A mechanism involving electron capture is suggested for the hAR activation. We propose that hAR inhibitors should not be designed against the native protein but against the activated form as obtained from X-ray irradiation. Furthermore, since the reactive species produced under irradiation conditions are the same as those produced under oxidative stress, the described irradiation method can be applied to other relevant proteins under oxidative stress environments.This work was started, and partly supported by a grant from the Spanish Nuclear Council (CSN)

Safety, feasibility, and hemodynamic response of regadenoson for stress perfusion CMR

Owing to its pharmacodynamics and posology, the use of regadenoson for stress cardiac magnetic resonance (CMR) has potential advantages over other vasodilators. We sought to evaluate the safety, hemodynamic response and diagnostic performance of regadenoson stress-CMR in routine clinical practice. All regadenoson stress-CMR examinations performed between May 2017 and July 2020 at our institution were retrospectively reviewed. A total of 698 studies were included for the final analysis. A conventional stress/rest protocol was performed using a 1.5T MRI scanner (Magnetom Aera, Siemens Healthineers, Erlangen, Germany). Adverse events, clinical symptoms, and hemodynamic response were assessed. Diagnostic accuracy of the test was evaluated in patients who underwent invasive coronary angiography. Nearly half of patients (48.5%) remained asymptomatic. Most common clinical symptoms included dyspnea (137, 19.6%), chest pain (116, 16.6%) and flushing (44, 6.3%). Two patients (0.28%) could not complete the examination due to severe hypotension or unbearable chest pain. Overall, an increase in heart rate (HR) response (36.2% [IQR: 22.5–50.9]) and a decrease in systolic and diastolic blood pressure (BP) (median systolic BP response of -5% [IQR: -11.5-0.6]; median diastolic BP response of -6.3 mmHg [IQR: -13.4-0]) was observed. Patients with symptoms induced by regadenoson showed higher HR response (40.3%, IQR: 26.4–56.1 vs. 32.4%, IQR: 19-45.6, p<0.001), whereas a blunted HR response was observed in diabetic (29.6%, IQR: 18.4–42 p<0.001), obese (31.7%, IQR: 20.7–46.2 p=0.005) and patients aged 70 years or older (32.9%, IQR: 22.6–43.1 p<0.001). Overall, regadenoson stress-CMR showed 95.65% (IQ 91.49–99.81) sensitivity, 54.84% (IQ 35.71–73.97) specificity, 86.99% (IQ 82.74–94.68) positive predictive value, and 77.27% (IQ 57.49–97.06) negative predictive value for detecting significant coronary stenosis as compared with invasive coronary angiography. Regadenoson is a well-tolerated vasodilator that can be safely employed for stress perfusion CMR, with high diagnostic performanc

Tratamiento de la insuficiencia cardíaca avanzada mediante estimulación biventricular. Experiencia inicial en una serie de 22 casos consecutivos

Recent data suggest that biventricular pacing may play an important role in treating advanced heart failure in the presence of a significant interventricular and/or intraventricular conduction disorder by correcting cardiac dysynchrony. In this article, we review the initial technical and clinical experience with cardiac resynchronization therapy in an electrophysiology laboratory. METHODS: The first 22 consecutive patients with severe congestive heart failure, ejection fraction < 0.35, NYHA functional class III or IV, and QRS duration > 120 ms who were implanted biventricular pacemakers were studied. Clinical, electrocardiographic, and echocardiographic evaluations were made before and three months after pacemaker implantation. Acute functional capacity testing with peak oxygen uptake was measured during biventricular pacing and during intrinsic rhythm or right ventricular pacing three months after the implantation procedure. RESULTS: The success rate of pacemaker implantation was 95%. Pre-discharge left ventricular pacing was achieved in 91%, with an average pacing threshold of 1.53 (1.04) volts. NYHA functional class improved (p = 0.039) from 3.4 (0.7) to 2.3 (0.78). The rate of hospitalization for heart failure decreased from an average of 3.12 (0.58) three months before the procedure to 1.38 (0.34) three months after the procedure. Peak oxygen uptake was significantly greater (p = 0.028) during biventricular pacing: 14.89 (2.1) ml/min/kg, than during intrinsic rhythm or right ventricular pacing: 12.65 (2.3) ml/min/kg. CONCLUSIONS: Cardiac resynchronization therapy can be performed safely and with a high success rate in the electrophysiology laboratory. Biventricular pacing seems to improve the symptoms of congestive heart failure in patients with evidence of atrioventricular and/or interventricular/intraventricular dysynchrony. An acute benefit in peak oxygen uptake was associated with biventricular pacing after the implantation procedure

Efecto de la localización del electrodo ventricular izquierdo sobre los parámetros ecocardiográficos de asincronía en pacientes sometidos a terapia de resincronización cardíaca

Introduction and objectives. Cardiac resynchronization therapy has been shown to be an option in the treatment of patients with congestive heart failure. The current indication for this treatment is based on clinical and electrocardiographic criteria, although echocardiography has also been shown to be a useful tool for the diagnosis of ventricular dyssynchrony. The aim of this study was to assess left ventricular dyssynchrony by echocardiography and to evaluate the effect of the stimulation site on the magnitude of resynchronization. Patients and method. We studied 25 patients with biventricular stimulation (left ventricular lead located in a lateral position in 13 patients, and in an anterior position in 12). A complete echo-Doppler evaluation, including left ventricular ejection fraction, ventricular diameters and parameters of inter- and intraventricular dyssynchrony, was performed before implantation and 3 months after the procedure, with the device connected and disconnected. Results. Left ventricular ejection fraction increased significantly from 23.7 (6.5) to 27.8 (5.5) (P=.007) at 3 months. In the group as a whole, biventricular pacing was associated with a significant decrease in all intraventricular dyssynchrony parameters (septal-to-lateral wall motion delay and septal-to-posterior wall motion delay). This decrease in septal-to-posterior wall motion delay and septalto- lateral wall motion delay was significantly greater in patients with the electrode implanted in the lateral position (58.1 ms vs 118 ms; P=.02) than with the lead in the anterior position (39.5 ms vs 86.5 ms; P=.04). Three patients, all with the electrode in an anterior location, were considered non-responders. Conclusions. Left lateral free wall stimulation provided significantly better intraventricular resynchronization compared to stimulation at an anterior site. Echocardiography is a useful tool to evaluate changes in intra- and interventricular synchrony related to the pacing site

Supramolecular gels: a versatile crystallization toolbox

Supramolecular gels are unique materials formed through the self-assembly of molecular building blocks, typically low molecular weight gelators (LMWGs), driven by non-covalent interactions. The process of crystallization within supramolecular gels has broadened the scope of the traditional gel-phase crystallization technique offering the possibility of obtaining crystals of higher quality and size. The broad structural diversity of LMWGs allows crystallization in multiple organic and aqueous solvents, favouring screening and optimization processes and the possibility to search for novel polymorphic forms. These supramolecular gels have been used for the crystallization of inorganic, small organic compounds of pharmaceutical interest, and proteins. Results have shown that these gels are not only able to produce crystals of high quality but also to influence polymorphism and physicochemical properties of the crystals, giving rise to crystals with potential new bio- and technological applications. Thus, understanding the principles of crystallization in supramolecular gels is essential for tailoring their properties and applications, ranging from drug delivery systems to composite crystals with tunable stability properties. In this review, we summarize the use of LMWG-based supramolecular gels as media to grow single crystals of a broad range of compounds

Non-conservation of folding rates in the thioredoxin family reveals degradation of ancestral unassisted-folding

Evolution involves not only adaptation, but also the degradation of superfluous features. Many examples of degradation at the morphological level are known (vestigial organs, for instance). However, the impact of degradation on molecular evolution has been rarely addressed. Thioredoxins serve as general oxidoreductases in all cells. Here, we report extensive mutational analyses on the folding of modern and resurrected ancestral bacterial thioredoxins. Contrary to claims from recent literature, in vitro folding rates in the thioredoxin family are not evolutionarily conserved, but span at least a ∼100-fold range. Furthermore, modern thioredoxin folding is often substantially slower than ancestral thioredoxin folding. Unassisted folding, as probed in vitro, thus emerges as an ancestral vestigial feature that underwent degradation, plausibly upon the evolutionary emergence of efficient cellular folding assistance. More generally, our results provide evidence that degradation of ancestral features shapes, not only morphological evolution, but also the evolution of individual proteins.This research was supported by FEDER Funds, grant BIO2015-66426-R from the Spanish Ministry of Economy and Competitiveness ( J.M.S.-R.), grant RGP0041/2017 from the Human Frontier Science Program ( J.M.S.-R. and E.A.G.) and National Institutes of Health 1R01AR069137 (E.A.G.), Department of Defence MURI W911NF-16-1-0372 (E.A.G.)