PREDICTORS OF ATRIAL FIBRILLATION RECURRENCE AFTER RADIOFREQUENCY ABLATION IN PATIENTS WITH CHRONIC HEART FAILURE.

OBJECTIVE: The aim: To study the predictive power of demographic, hemodynamic and electrocardiographic factors for atrial fibrillation recurrence after radiofrequency ablation (RFA) in patients with chronic heart failure. PATIENTS AND METHODS: Materials and methods: Study included 120 patients, aged 59,80±10,08 years old with chronic heart failure with preserved left ventricular ejection fraction who were undergo RFA due to atrial fibrillation (AF). A standard 12-lead electrocardiogram (ECG) was registered. Before the procedure the standard echocardiographic parameters were obtained. After 12 months, patients were divided into 2 groups: non-recurrence group and recurrence group. RESULTS: Results: As a result of prospective follow-up within 12 months AF recurrences were observed in 32 patients (27%), 88 patients remained non-recurrent (73%). The left atrium (LA) diameter and aortic root diameter were lager in the recurrence group (4,59±0,45 vs. 4,08±0,61 cm, p<0,001; 3,37±0,60 vs. 2,80±0,67 cm, p<0,001). The AF recurrence group before RFA had a significantly longer QTc interval than the non-recurrence group (387,23±2,31 vs. 341,22±8,91 ms, p<0,010). ROC curve analysis revealed LA diameter the most sensitive factor for AF recurrence after RFA. CONCLUSION: Conclusions: QTc duration before radiofrequency ablation and its prolongation after intervention are independent predictors of atrial fibrillation recurrence; left atrium diameter before ablation is a highly sensitive predictor in patients with chronic heart failure with preserved left ventricular ejection fraction.

Bailout Deep Septal LV Pacing to Treat Inadvertent Complete AV Block During Complex Ablation Procedure.

We present a case of persistent complete atrioventricular block that occurred during the diagnostic portion of a premature ventricular contractions' radiofrequency ablation in a complex heart failure patient. The case was managed by bailout deep left ventricular septal pacing after bipolar radiofrequency elimination of premature ventricular contractions.

E. coli Expression and Purification of Microbial and Viral Rhodopsins.

Microbial rhodopsins have become an indispensable tool for neurobiology. Of thousands of identified microbial rhodopsins, a minute fraction has been studied so far and they have shown remarkable functional diversity suggesting more great promises that this large family holds. Effective production of recombinant microbial and viral rhodopsins is a prerequisite for the success of functional and structural studies of these proteins. Escherichia coli (E. coli) are suitable for high yield expression of many of microbial and viral rhodopsins and they facilitate rapid exploration of this large protein family.

Detection of cancerous cervical cells using physical adhesion of fluorescent silica particles and centripetal force.

Here we describe a non-traditional method to identify cancerous human cervical epithelial cells in a culture dish based on physical adhesion between silica beads and cells. It is a simple optical fluorescence-based technique which detects the relative difference in the amount of fluorescent silica beads physically adherent to surfaces of cancerous and normal cervical cells. The method utilizes the centripetal force gradient that occurs in a rotating culture dish. Due to the variation in the balance between adhesion and centripetal forces, cancerous and normal cells demonstrate clearly distinctive distributions of the fluorescent particles adherent to the cell surface over the culture dish. The method demonstrates higher adhesion of silica particles to normal cells compared to cancerous cells. The difference in adhesion was initially observed by atomic force microscopy (AFM). The AFM data were used to design the parameters of the rotational dish experiment. The optical method that we describe is much faster and technically simpler than AFM. This work provides proof of the concept that physical interactions can be used to accurately discriminate normal and cancer cells.

His-optimized cardiac resynchronization therapy in a patient with heart failure and right bundle branch block: a case report.

BACKGROUND: Cardiac resynchronization therapy (CRT) is an option for treatment for chronic heart failure (HF) associated with left bundle branch block (LBBB). Patients with HF and right bundle branch block (RBBB) have potentially worse outcomes in comparison to LBBB. Traditional CRT in RBBB can increase mortality and HF deterioration rates over native disease progression. His bundle pacing may improve the results of CRT in those patients. Furthermore, atrioventricular node ablation (AVNA) for rate control in atrial fibrillation (AF) can be challenging in patients with previously implanted leads in His region. CASE SUMMARY: We report the case of 74-year-old gentleman with a 5-year history of HF, permanent AF with a rapid ventricular response, and RBBB. He was admitted to the hospital with complaints of severe weakness and shortness of breath. Left ventricular ejection fraction (LVEF) was decreased (41%), right ventricle (RV) was dilated (41 mm), and QRS was prolonged (200 ms) with RBBB morphology. The patient underwent His-optimized CRT with further left-sided AVNA. As a result, LVEF increased to 51%, RV dimensions decreased to 35 mm with an improvement of the clinical status during a 6-month follow-up. DISCUSSION: Patients with AF, RBBB, and HF represent the least evaluated clinical subgroup of individuals with less beneficial clinical outcomes according to CRT studies. Achieving the most effective resynchronization could require pacing fusion from sites beyond traditional with the intention to recruit intrinsic conduction pathways. This approach can be favourable for reducing RV dilatation, improving LVEF, and maximizing electrical resynchronization.

Molecular model of a sensor of two-component signaling system.

Two-component systems (TCS) are widespread signaling systems present in all domains of life. TCS typically consist of a signal receptor/transducer and a response regulator. The receptors (histidine kinases, chemoreceptors and photoreceptors) are often embedded in the membrane and have a similar modular structure. Chemoreceptors were shown to function in highly ordered arrays, with trimers of dimers being the smallest functional unit. However, much less is known about photoreceptors. Here, we use small-angle scattering (SAS) to show that detergent-solubilized sensory rhodopsin II in complex with its cognate transducer forms dimers at low salt concentration, which associate into trimers of dimers at higher buffer molarities. We then fit an atomistic model of the whole complex into the SAS data. The obtained results suggest that the trimer of dimers is "tripod"-shaped and that the contacts between the dimers occur only through their cytoplasmic regions, whereas the transmembrane regions remain unconnected.

Ultrabright fluorescent mesoporous silica nanoparticles.

The first successful approach to synthesizing ultrabright fluorescent mesoporous silica nanoparticles is reported. Fluorescent dye is physically entrapped inside nanochannels of a silica matrix created during templated sol-gel self-assembly. The problem of dye leakage from open channels is solved by incorporation of hydrophobic groups in the silica matrix. This makes the approach compatible with virtually any dye that can withstand the synthesis. The method is demonstrated using the dye Rhodamine 6G. The obtained 40-nm silica particles are about 30 times brighter than 30-nm coated water-soluble quantum dots. The particles are substantially more photostable than the encapsulated organic dye itself.

Towards understanding of shape formation mechanism of mesoporous silica particles.

Growth of even simple crystals is a rather hard problem to describe because of the non-equilibrium nature of the process. Meso(nano)porous silica particles, which are self-assembled in a sol-gel template synthesis, demonstrate an example of shapes of high complexity, similar to those observed in the biological world. Despite such complexity, here we present the evidence that at least a part of the formation of these shapes is an equilibrium process. We demonstrate it for an example of mesoporous fibers, one of the abundant shapes. We present a quantitative proof that the fiber free energy is described by the Boltzmann distribution, which is predicted by the equilibrium thermodynamics. This finding may open up new ground for a quantitative description of the morphogenesis of complex self-assembled shapes, including biological hierarchy.

Molecular mechanism of light-driven sodium pumping.

The light-driven sodium-pumping rhodopsin KR2 from Krokinobacter eikastus is the only non-proton cation active transporter with demonstrated potential for optogenetics. However, the existing structural data on KR2 correspond exclusively to its ground state, and show no sodium inside the protein, which hampers the understanding of sodium-pumping mechanism. Here we present crystal structure of the O-intermediate of the physiologically relevant pentameric form of KR2 at the resolution of 2.1 Å, revealing a sodium ion near the retinal Schiff base, coordinated by N112 and D116 of the characteristic NDQ triad. We also obtained crystal structures of D116N and H30A variants, conducted metadynamics simulations and measured pumping activities of putative pathway mutants to demonstrate that sodium release likely proceeds alongside Q78 towards the structural sodium ion bound between KR2 protomers. Our findings highlight the importance of pentameric assembly for sodium pump function, and may be used for rational engineering of enhanced optogenetic tools.

Viral rhodopsins 1 are an unique family of light-gated cation channels.

Phytoplankton is the base of the marine food chain as well as oxygen and carbon cycles and thus plays a global role in climate and ecology. Nucleocytoplasmic Large DNA Viruses that infect phytoplankton organisms and regulate the phytoplankton dynamics encompass genes of rhodopsins of two distinct families. Here, we present a functional and structural characterization of two proteins of viral rhodopsin group 1, OLPVR1 and VirChR1. Functional analysis of VirChR1 shows that it is a highly selective, Na+/K+-conducting channel and, in contrast to known cation channelrhodopsins, it is impermeable to Ca2+ ions. We show that, upon illumination, VirChR1 is able to drive neural firing. The 1.4 Å resolution structure of OLPVR1 reveals remarkable differences from the known channelrhodopsins and a unique ion-conducting pathway. Thus, viral rhodopsins 1 represent a unique, large group of light-gated channels (viral channelrhodopsins, VirChR1s). In nature, VirChR1s likely mediate phototaxis of algae enhancing the host anabolic processes to support virus reproduction, and therefore, might play a major role in global phytoplankton dynamics. Moreover, VirChR1s have unique potential for optogenetics as they lack possibly noxious Ca2+ permeability.

Logic networks based on immunorecognition processes.

The biochemical system logically processing biochemical signals using immune-specific and biocatalytic reactions was designed, and the generated output signals were analyzed by AFM and optical means. Different patterns of immune signals resulted in the formation of various interfacial structures followed by biocatalytic reactions activated by the next set of biochemical inputs. The developed approach to multisignal biosensing allows qualitative evaluation of the biochemical information in terms of YES-NO, providing the base for novel molecular-level logic analysis of complex patterns of biochemical signals. Application of AFM to read out the structures generated on the interface could potentially lead to substantial miniaturization of the immune logic systems.

Cellular energetics and mitochondrial uncoupling in canine aging.

The first domesticated companion animal, the dog, is currently represented by over 190 unique breeds. Across these numerous breeds, dogs have exceptional variation in lifespan (inversely correlated with body size), presenting an opportunity to discover longevity-determining traits. We performed a genome-wide association study on 4169 canines representing 110 breeds and identified novel candidate regulators of longevity. Interestingly, known functions within the identified genes included control of coat phenotypes such as hair length, as well as mitochondrial properties, suggesting that thermoregulation and mitochondrial bioenergetics play a role in lifespan variation. Using primary dermal fibroblasts, we investigated mitochondrial properties of short-lived (large) and long-lived (small) dog breeds. We found that cells from long-lived breeds have more uncoupled mitochondria, less electron escape, greater respiration, and capacity for respiration. Moreover, our data suggest that long-lived breeds have higher rates of catabolism and ß-oxidation, likely to meet elevated respiration and electron demand of their uncoupled mitochondria. Conversely, cells of short-lived (large) breeds may accumulate amino acids and fatty acid derivatives, which are likely used for biosynthesis and growth. We hypothesize that the uncoupled metabolic profile of long-lived breeds likely stems from their smaller size, reduced volume-to-surface area ratio, and therefore a greater need for thermogenesis. The uncoupled energetics of long-lived breeds lowers reactive oxygen species levels, promotes cellular stress tolerance, and may even prevent stiffening of the actin cytoskeleton. We propose that these cellular characteristics delay tissue dysfunction, disease, and death in long-lived dog breeds, contributing to canine aging diversity.

Unique structure and function of viral rhodopsins.

Recently, two groups of rhodopsin genes were identified in large double-stranded DNA viruses. The structure and function of viral rhodopsins are unknown. We present functional characterization and high-resolution structure of an Organic Lake Phycodnavirus rhodopsin II (OLPVRII) of group 2. It forms a pentamer, with a symmetrical, bottle-like central channel with the narrow vestibule in the cytoplasmic part covered by a ring of 5 arginines, whereas 5 phenylalanines form a hydrophobic barrier in its exit. The proton donor E42 is placed in the helix B. The structure is unique among the known rhodopsins. Structural and functional data and molecular dynamics suggest that OLPVRII might be a light-gated pentameric ion channel analogous to pentameric ligand-gated ion channels, however, future patch clamp experiments should prove this directly. The data shed light on a fundamentally distinct branch of rhodopsins and may contribute to the understanding of virus-host interactions in ecologically important marine protists.

Synthesis of ultrabright nanoporous fluorescent silica discoids using an inorganic silica precursor.

The templated sol-gel synthesis of ultrabright fluorescent nanoporous silica particles based on the use of organic silica sources has previously been reported. The use of organosilanes as the main silica precursors has a number of issues, in particular, the low robustness of the synthesis due to instability of the organic silica source. Here we report on a novel synthesis of ultrabright fluorescent nanoporous silica discoids (a specific shape in-between the sphere and disk) of 3.1 ± 0.7 microns in size, which were prepared using a stable inorganic sodium silicate silica source. Organic fluorescent dye Rhodamine 6G (R6G) was physically (non-covalently) entrapped inside cylindrical nanochannels of ∼4-5 nm in diameter. In contrast to the synthesis with organic silica precursors, the obtained particles showed an excessive leakage of dye. To prevent this leakage, we modified the synthesis by adding a small amount of a secondary silica source. The synthesized particles show virtually no leakage, high photostability, and a brightness equivalent to the fluorescence of up to 7 × 10(7) free R6G molecules. This is about 7 times higher than the fluorescent brightness of particles of the same size made of CdSe/ZnS quantum dots, and 420 times higher than the brightness of the same volume of aqueous solution of free R6G dye.