Mapeamento do Potencial da Superfície Corporal durante a Despolarização Ventricular em Atletas com Intervalo PQ Prolongado após Exercício / Body Surface Potential Mapping during Ventricular Depolarization in Athletes with Prolonged PQ Interval after Exercise

Resumo Fundamento: O prolongamento do intervalo PQ, geralmente associado a um atraso na condução atrioventricular, pode estar relacionado a alterações na propagação do impulso intraventricular. Objetivo: Avaliar, por meio do mapeamento do potencial de superfície corporal (BSPM), o processo de despolarização ventricular em atletas com intervalos PQ prolongados em repouso e após o exercício. Métodos: O estudo incluiu 7 esquiadores cross-country com intervalo PQ superior a 200 ms (grupo PQ Prolongado) e 7 com intervalo PQ inferior a 200 ms (grupo PQ Normal). O BSPM de 64 derivações unipolares do tronco foi realizado antes (Pré-Ex) e após o teste ergométrico de bicicleta (Pós-Ex). Mapas equipotenciais da superfície corporal foram analisados durante a despolarização ventricular. O nível de significância foi de 5%. Resultados: Comparado com atletas com PQ Normal, o primeiro e o segundo períodos de posição estável dos potenciais cardíacos na superfície do tronco foram mais longos, e a formação da distribuição de potencial "sela" ocorreu mais tarde, no Pré-Ex, nos atletas com PQ Prolongado. No Pós-Ex, o grupo PQ Prolongado apresentou um encurtamento do primeiro e segundo períodos de distribuições de potencial estáveis e uma diminuição no tempo de aparecimento do fenômeno "sela" em relação ao Pré-Ex (para valores próximos aos do Normal -Grupo PQ). Além disso, no Pós-Ex, a primeira inversão das distribuições de potencial e a duração total da despolarização ventricular em atletas com PQ Prolongado diminuíram em comparação com o Pré-Ex e com valores semelhantes em atletas com PQ Normal. Em comparação com atletas com PQ Normal, a segunda inversão foi mais longa no Pré-Ex e Pós-Ex em atletas com PQ Prolongado. Conclusão: Atletas com PQ prolongado apresentaram diferenças significativas nas características temporais do BSPM durante a despolarização ventricular, tanto em repouso quanto após o exercício, em comparação com atletas com PQ normal.

Abstract Background: Prolongation of the PQ interval, generally associated with an atrioventricular conduction delay, may be related to changes in intraventricular impulse spreading. Objective: To assess, using body surface potential mapping (BSPM), the process of ventricular depolarization in athletes with prolonged PQ intervals at rest and after exercise. Methods: The study included 7 cross-country skiers with a PQ interval of more than 200 ms (Prolonged-PQ group) and 7 with a PQ interval of less than 200 ms (Normal-PQ group). The BSPM from 64 unipolar torso leads was performed before (Pre-Ex) and after the bicycle exercise test (Post-Ex). Body surface equipotential maps were analyzed during ventricular depolarization. The significance level was 5%. Results: Compared to Normal-PQ athletes, the first and second periods of the stable position of cardiac potentials on the torso surface were longer, and the formation of the "saddle" potential distribution occurred later, at Pre-Ex, in Prolonged-PQ athletes. At Post-Ex, the Prolonged-PQ group showed a shortening of the first and second periods of stable potential distributions and a decrease in appearance time of the "saddle" phenomenon relative to Pre-Ex (to the values near to those of the Normal-PQ group). Additionally, at Post-Ex, the first inversion of potential distributions and the total duration of ventricular depolarization in Prolonged-PQ athletes decreased compared to Pre-Ex and with similar values in Normal-PQ athletes. Compared to Normal-PQ athletes, the second inversion was longer at Pre-Ex and Post-Ex in Prolonged-PQ athletes. Conclusion: Prolonged-PQ athletes had significant differences in the temporal characteristics of BSPM during ventricular depolarization both at rest and after exercise as compared to Normal-PQ athletes.

Glycosylation and charge distribution orchestrates the conformational ensembles of a biotechnologically promissory phytase in different pHs - a computational study.

Phytases [myo-inositol(1,2,3,4,5,6) hexakisphosphate phosphohydrolases] are phytate-specific phosphatases not present in monogastric animals. Nevertheless, they are an essential supplement to feeding such animals and for human special diets. It is crucial, hence, the biotechnological use of phytases with intrinsic stability and activity at the acid pHs from gastric environments. Here we use Metadynamics (METADY) simulations to probe the conformational space of the Aspergillus nidulans phytase and the differential effects of pH and glycosylation in this same space. The results suggest that strategic combinations of pH and glycosylation affect the stability of native-like conformations and alternate these structures from a metastable to a stable profile. Furthermore, the protein segments previously reported as more thermosensitive in phytases from this family present a pivotal role in the conformational changes at different conditions, especially H2, H5-7, L8, L10, L12, and L17. Also, the glycosylations and the pH-dependent charge balance modulate the mobility and interactions at these same regions, with consequences for the surface solvation and active site exposition. Finally, although the glycosylations have stabilized the native structure and improved the substrate docking at all the studied pHs, the data suggest a higher phytate receptivity at catalytic poses for the unglycosylated structure at pH 6.5 and the glycosylated one at pH 4.5. This behavior agrees with the exact change in optimum pH reported for this enzyme, expressed on low or high glycosylating systems. We hope the results and insights presented here will be helpful in future approaches for rational engineering of technologically promising phytases and intelligent planning of their heterologous expression systems and conditions for use.Communicated by Ramaswamy H. Sarma.

Mapeamento Potencial de Superfície Corporal durante a Despolarização Ventricular em Ratos Após Exercício Exaustivo Agudo / Body Surface Potential Mapping during Ventricular Depolarization in Rats after Acute Exhaustive Exercise

Resumo Fundamento O exercício físico exaustivo pode causar alterações significantes nas propriedades elétricas do miocárdio. Objetivo Avaliar, através do mapeamento potencial de superfície corporal, a atividade elétrica do coração de ratos durante a despolarização ventricular após exercício exaustivo agudo. Métodos Ratos machos com doze semanas de idade foram submetidos a exercício agudo em esteira a 36 m/min até a exaustão. Eletrocardiogramas unipolares (ECGs) da superfície do tronco foram registrados em ratos anestesiados com zoletil três a cinco dias antes (Pré-Ex), 5 e 10 minutos após exercício exaustivo (Pós-Ex 5 e Pós-Ex 10, respectivamente) simultaneamente com ECGs nas derivações dos membros. Os mapas potenciais de superfície corporal instantâneos (BSPMs, body surface potential maps ) foram analisados durante a despolarização ventricular. Os valores de p <0,05 foram considerados estatisticamente significantes. Resultados Comparado com o Pré-Ex, uma conclusão precoce da segunda inversão de distribuições de potencial, uma conclusão precoce da despolarização ventricular, bem como uma diminuição na duração da fase média e a duração total da despolarização ventricular nos BSPMs foram reveladas no Pós-Ex5. Além disso, em comparação com o Pré-Ex, um aumento na amplitude do extremo negativo do BSPM no pico da onda R no ECG na derivação II (pico RII) e uma diminuição na amplitude do extremo negativo do BSPM a 3 e 4 ms após o pico RII foram demonstrados no Pós-Ex 5. No Pós-Ex 10, os parâmetros dos BSPMs não diferiram daqueles do Pré-Ex. Conclusão Em ratos, o exercício exaustivo agudo causa alterações reversíveis nas características temporais e de amplitude dos BSPMs durante a despolarização ventricular, provavelmente relacionadas a alterações na excitação da massa principal do miocárdio ventricular.

Abstract Background Exhaustive physical exercise can cause substantial changes in the electrical properties of the myocardium. Objective To evaluate, using body surface potential mapping, the electrical activity of the heart in rats during ventricular depolarization after acute exhaustive exercise. Methods Twelve-week-old male rats were submitted to acute treadmill exercise at 36 m/min until exhaustion. Unipolar electrocardiograms (ECGs) from the torso surface were recorded in zoletil-anesthetized rats three to five days before (Pre-Ex), 5 and 10 minutes after exhaustive exercise (Post-Ex 5 and Post-Ex 10, respectively) simultaneously with ECGs in limb leads. The instantaneous body surface potential maps (BSPMs) were analyzed during ventricular depolarization. P values <0.05 were considered statistically significant. Results Compared with Pre-Ex, an early completion of the second inversion of potential distributions, an early completion of ventricular depolarization, as well as a decrease in the duration of the middle phase and the total duration of ventricular depolarization on BSPMs were revealed at Post-Ex 5. Also, compared with Pre-Ex, an increase in the amplitude of negative BSPM extremum at the R-wave peak on the ECG in lead II (RII-peak) and a decrease in the amplitude of negative BSPM extremum at 3 and 4 ms after RII-peak were showed at Post-Ex 5. At Post-Ex 10, parameters of BSPMs did not differ from those at Pre-Ex. Conclusion In rats, acute exhaustive exercise causes reversible changes in the temporal and amplitude characteristics of BSPMs during ventricular depolarization, most likely related to alterations in the excitation of the main mass of the ventricular myocardium.

Characterization of atrial arrhythmias in body surface potential mapping: A computational study.

PURPOSE: Atrial tachycardia (AT), flutter (AFL) and fibrillation (AF) are very common cardiac arrhythmias and are driven by localized sources that can be ablation targets. Non-invasive body surface potential mapping (BSPM) can be useful for early diagnosis and ablation planning. We aimed to characterize and differentiate the arrhythmic mechanisms behind AT, AFL and AF from the BSPM perspective using basic features reflecting their electrophysiology. METHODS: 19 simulations of 567-lead BSPMs were used to obtain dominant frequency (DF) maps and estimate the atrial driving frequencies using the highest DF (HDF). Regions with |DF-HDF|≤1Hz were segmented and characterized (size, area); the spatial distribution of the differences |DF-atrialHDFestimate| was qualitatively analyzed. Phase singularity points (SPs) were detected on maps generated with Hilbert transform after band-pass filtering around the HDF (±1Hz). Connected SPs along time (filaments) and their histogram (heatmaps) were used for rotational activity characterization (duration, spatiotemporal stability). Results were reproduced in clinical layouts (252 to 12 leads) and with different rotations and translations of the atria within the torso, and compared with the original 567-lead outcomes using structural similarity index (SSIM) between maps, sensitivity and precision in SP detection and direct feature comparison. Random forest and least-square based algorithms were used to classify the arrhythmias and their mechanisms' location, respectively, based on the obtained features. RESULTS: Frequency and phase analyses revealed distinct behavior between arrhythmias. AT and AFL presented uniform DF maps with low variance, while AF maps were more heterogeneous. Lower differences from the atrial HDF regions correlated with the driver location. Rotational activity was most stable in AFL, followed by AT and AF. Features were robust to lower spatial resolution layouts and modifications in the atrial geometry; DF and heatmaps presented decreasing SSIM along the layouts. The classification of the arrhythmias and their mechanisms' location achieved balanced accuracy of 72.0% and 73.9%, respectively. CONCLUSION: Non-invasive characterization of AT, AFL and AF based on realistic models highlights intrinsic differences between the arrhythmias, enhancing the BSPM utility as an auxiliary clinical tool.

Interaction of cysteine and its derivatives with monolayers of dipalmitoylphosphatidylcholine.

Molecular interactions between l-cysteine (Cys) and its ester derivatives (Cysx); l-cysteine ethyl ester (CE), l-cysteine methyl ester (CM) and N-acetyl l-cysteine (NAC) with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) monolayers were investigated using Langmuir film balance technique. The effect of charge on monolayers made of cysteine and three ester derivatives with DPPC was investigated by working with un-buffered and buffered subphases. Also, the effects of cysteine derivatives interaction with DPPC monolayers were studied measuring the change in the surface tension upon aminoacid injection in the subphase whilst keeping lipid molecular density and lateral packing controlled. Cysteine and its ester derivatives showed interfacial activity reducing the air/water surface tension (πi) by 4 mN m-1. However, ester derivatives were able to insert into preformed DPPC monolayers at much higher surface pressures (Δπ), indicating a preferential interaction of Cysx with DPPC. The results indicate that, although the different derivatives of cysteine presented low surface activity, they were able to favourably interact with DPPC monolayers. Also, compression isotherms experiments in binary mixtures indicate that the more surface active compounds stabilized the gel phase of DPPC. The charge on cysteine and its derivatives did not increase the observed effects.

Calculation of VS,max and Its Use as a Descriptor for the Theoretical Calculation of pKa Values for Carboxylic Acids.

The theoretical calculation of pKa values for Brønsted acids is a challenging task that involves sophisticated and time-consuming methods. Therefore, heuristic approaches are efficient and appealing methodologies to approximate these values. Herein, we used the maximum surface electrostatic potential (VS,max) on the acidic hydrogen atoms of carboxylic acids to describe the H-bond interaction with water (the same descriptor that is used to characterize σ-bonded complexes) and correlate the results with experimental pKa values to obtain a predictive model for other carboxylic acids. We benchmarked six different methods, all including an implicit solvation model (water): Five density functionals and the Møller⁻Plesset second order perturbation theory in combination with six different basis sets for a total of thirty-six levels of theory. The ωB97X-D/cc-pVDZ level of theory stood out as the best one for consistently reproducing the reported pKa values, with a predictive power of 98% correlation in a test set of ten other carboxylic acids.

The many faces (and phases) of ceramide and sphingomyelin I - single lipids.

Ceramides, the simplest kind of two-chained sphingolipids, contain a single hydroxyl group in position 1 of the sphingoid base. Sphingomyelins further contain a phosphocholine group at the OH of position 1 of ceramide. Ceramides and sphingomyelins show a variety of species depending on the fatty acyl chain length, hydroxylation, and unsaturation. Because of the relatively high transition temperature of sphingomyelin compared to lecithin and, particularly, of ceramides with 16:0-18:0 saturated chains, a widespread idea on their functional importance refers to formation of rather solid domains enriched in sphingomyelin and ceramide. Frequently, and especially in the cell biology field, these are generally (and erroneously) assumed to occur irrespective on the type of N-acyl chain in these lipids. This is because most studies indicating such condensed ordered domains employed sphingolipids with acyl chains with 16 carbons while scarce attention has been focused on the influence of the N-acyl chain on their surface properties. However, abundant evidence has shown that variations of the N-acyl chain length in ceramides and sphingomyelins markedly affect their phase state, interfacial elasticity, surface topography, electrostatics and miscibility and that, even the usually conceived "condensed" sphingolipids and many of their mixtures, may exhibit liquid-like expanded states. This review is a summarized overview of our work and of related others on some facts regarding membranes composed of single molecular species of ceramide and sphingomyelin. A second part is dedicated to discuss the miscibility properties between species of sphingolipids that differ in N-acyl and oligosaccharide chains.

Computational models for monitoring the trans-membrane potential with fluorescent probes: the DiSC3(5) case.

Fluorescent permeant charged probes are commonly used for monitoring the trans-membrane potential in lipid vesicles and biological membranes, which has been earlier described by various mathematical models. In the present study, we developed a more complex model based on the computational step-by-step analysis of the influence of various factors, such as the membrane surface potential, ionic strength, and the aggregation properties of cationic cyanine probe DiSC3(5) in the membrane and aqueous phases, in addition to the Nernstian distribution of the probe across the membrane and the hydrophobic interaction with the lipid bilayer. The final full model allows prediction of the optimal experimental conditions for monitoring the trans-membrane potential, such as the probe/lipid ratio and the concentration of liposomes, with a given percentage of negatively charged phospholipids in the membrane, the ionic strength of the aqueous media, the "membrane-water" partition coefficient and the aggregation properties of the probe, as well as the most adequate mode of fluorescence measurement. In agreement with many experimental studies, this model showed high voltage sensitivity of the quantity of the aqueous phase DiSC3(5) monomers, showing its almost exponential decrease with an increase in the trans-membrane potential value. The model also demonstrated the highest voltage sensitivity of the ratio of the quantity of DiSC3(5) monomers in the aqueous phases to that in the membrane phase. A new combined parameter, the logarithmic function of this ratio, demonstrated almost linear changes within a wide range of the trans-membrane potential changes.

Atypical surface behavior of ceramides with nonhydroxy and 2-hydroxy very long-chain (C28-C32) PUFAs.

Unique species of ceramide (Cer) with very-long-chain polyunsaturated fatty acid (VLCPUFA), mainly 28-32 carbon atoms, 4-5 double bonds, in nonhydroxy and 2-hydroxy forms (n-V Cer and h-V Cer, respectively), are generated in rat spermatozoa from the corresponding sphingomyelins during the acrosomal reaction. The aim of this study was to determine the properties of these sperm-distinctive ceramides in Langmuir monolayers. Individual Cer species were isolated by HPLC and subjected to analysis of surface pressure, surface potential, and Brewster angle microscopy (BAM) as a function of molecular packing. In comparison with known species of Cer, n-V Cer and h-V Cer species showed much larger mean molecular areas and increased molecular dipole moments in liquid expanded phases, which suggest bending and partial hydration of the double bonded portion of the VLCPUFA. The presence of the 2-hydoxyl group induced a closer molecular packing in h-V Cer than in their chain-matched n-V Cer. In addition, all these Cer species showed liquid-expanded to liquid-condensed transitions at room temperature. Existence of domain segregation was confirmed by BAM. Additionally, thermodynamic analysis suggests a phase transition close to the physiological temperature for VLCPUFA-Cers if organized as bulk dispersions.

Improved relationship between left and right ventricular electrical activation after cardiac resynchronization therapy in heart failure patients can be quantified by body surface potential mapping

OBJECTIVES: Few studies have evaluated cardiac electrical activation dynamics after cardiac resynchronization therapy. Although this procedure reduces morbidity and mortality in heart failure patients, many approaches attempting to identify the responders have shown that 30% of patients do not attain clinical or functional improvement. This study sought to quantify and characterize the effect of resynchronization therapy on the ventricular electrical activation of patients using body surface potential mapping, a noninvasive tool. METHODS: This retrospective study included 91 resynchronization patients with a mean age of 61 years, left ventricle ejection fraction of 28%, mean QRS duration of 182 ms, and functional class III/IV (78%/22%); the patients underwent 87-lead body surface mapping with the resynchronization device on and off. Thirty-six patients were excluded. Body surface isochronal maps produced 87 maximal/mean global ventricular activation times with three regions identified. The regional activation times for right and left ventricles and their inter-regional right-to-left ventricle gradients were calculated from these results and analyzed. The Mann-Whitney U-test and Kruskall-Wallis test were used for comparisons, with the level of significance set at p≤0.05. RESULTS: During intrinsic rhythms, regional ventricular activation times were significantly different (54.5 ms vs. 95.9 ms in the right and left ventricle regions, respectively). Regarding cardiac resynchronization, the maximal global value was significantly reduced (138 ms to 131 ms), and a downward variation of 19.4% in regional-left and an upward variation of 44.8% in regional-right ventricular activation times resulted in a significantly reduced inter-regional gradient (43.8 ms to 17 ms). CONCLUSIONS: Body surface potential mapping in resynchronization patients yielded electrical ventricular ...

Ascorbyl palmitate interaction with phospholipid monolayers: electrostatic and rheological preponderancy.

Ascorbyl palmitate (ASC16) is an anionic amphiphilic molecule of pharmacological interest due to its antioxidant properties. We found that ASC16 strongly interacted with model membranes. ASC16 penetrated phospholipid monolayers, with a cutoff near the theoretical surface pressure limit. The presence of a lipid film at the interface favored ASC16 insertion compared with a bare air/water surface. The adsorption and penetration time curves showed a biphasic behavior: the first rapid peak evidenced a fast adsorption of charged ASC16 molecules to the interface that promoted a lowering of surface pH, thus partially neutralizing and compacting the film. The second rise represented an approach to the equilibrium between the ASC16 molecules in the subphase and the surface monolayer, whose kinetics depended on the ionization state of the film. Based on the Langmuir dimiristoylphosphatidylcholine+ASC16 monolayer data, we estimated an ASC16 partition coefficient to dimiristoylphosphatidylcholine monolayers of 1.5×10(5) and a ΔGp=-6.7kcal·mol(-1). The rheological properties of the host membrane were determinant for ASC16 penetration kinetics: a fluid membrane, as provided by cholesterol, disrupted the liquid-condensed ASC16-enriched domains and favored ASC16 penetration. Subphase pH conditions affected ASC16 aggregation in bulk: the smaller structures at acidic pHs showed a faster equilibrium with the surface film than large lamellar ones. Our results revealed that the ASC16 interaction with model membranes has a highly complex regulation. The polymorphism in the ASC16 bulk aggregation added complexity to the equilibrium between the surface and subphase form of ASC16, whose understanding may shed light on the pharmacological function of this drug.