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Genetic testing is crucial in inherited arrhythmogenic channelopathies; however, the clinical interpretation of genetic variants remains challenging. Incomplete penetrance, oligogenic, polygenic or multifactorial forms of channelopathies further complicate variant interpretation. We identified the KCNQ1/p.D446E variant in 2/63 patients with long QT syndrome, 30-fold more frequent than in public databases. We thus characterized the biophysical phenotypes of wildtype and mutant IKs co-expressing these alleles with the ß-subunit minK in HEK293 cells. KCNQ1 p.446E homozygosity significantly shifted IKs voltage dependence to hyperpolarizing potentials in basal conditions (gain of function) but failed to shift voltage dependence to hyperpolarizing potentials (loss of function) in the presence of 8Br-cAMP, a protein kinase A activator. Basal IKs activation kinetics did not differ among genotypes, but in response to 8Br-cAMP, IKs 446 E/E (homozygous) activation kinetics were slower at the most positive potentials. Protein modeling predicted a slower transition of the 446E Kv7.1 tetrameric channel to the stabilized open state. In conclusion, biophysical and modelling evidence shows that the KCNQ1 p.D446E variant has complex functional consequences including both gain and loss of function, suggesting a contribution to the pathogenesis of arrhythmogenic phenotypes as a functional risk allele.

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Experimental studies of TiO2 nanotubes have been conducted for nearly three decades and have revealed the remarkable advantages of this material. Research based on computer simulations is much rarer, with research using density functional theory (DFT) being the most significant in this field. It should be noted, however, that this approach has significant limitations when studying the macroscopic properties of nanostructures such as nanosheets and nanotubes. An alternative with great potential has emerged: classical molecular dynamics simulations (MD). MD Simulations offer the possibility to study macroscopic properties such as the density of phonon states (PDOS), power spectra, infrared spectrum, water absorption and others. From this point of view, the present study focuses on the distinction between the phases of anatase and rutile TiO2. The LAMMPS package is used to study both the structural properties by applying the radial distribution function (RDF) and the electromagnetic properties of these phases. Our efforts are focused on exploring the effect of temperature on the vibrational properties of TiO2 anatase nanotubes and an in-depth analysis of how the phononic softening phenomenon affects TiO2 nanostructures to improve the fundamental understanding in different dimensions and morphological configurations. A careful evaluation of the stability of TiO2 nanolamines and nanotubes at different temperatures is performed, as well as the adsorption of water on the nanosurface of TiO2, using three different water models.

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Low-frequency mutations associated with drug resistance have been related to virologic failure in subjects with no history of pre-treatment and recent HIV diagnosis. In total, 78 antiretroviral treatment (ART)-naïve subjects with a recent HIV diagnosis were selected and followed by CD4+ T lymphocytes and viral load tests to detect virologic failure. We sequenced the basal samples retrospectively using next-generation sequencing (NGS), looking for low-frequency mutations that had not been detected before using the Sanger sequencing method (SSM) and describing the response to ART. Twenty-two subjects developed virologic failure (VF), and thirteen of them had at least one drug-resistance mutation associated with Reverse Transcriptase Inhibitors (RTI) and Protease Inhibitors (PIs) at frequency levels ≤ 1%, not detected previously in their basal genotyping test. No resistance mutations were observed to Integrase Strand Transfer Inhibitors (INSTIs). We identified a possible cause of VF in ART-naïve subjects with low-frequency mutations detected. To our knowledge, this is the first evaluation of pre-existing drug resistance for HIV-1 minority variants carried out on ART-naïve people living with HIV/AIDS (PLWHA) by analyzing the HIV-1 pol gene using NGS in the country.

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BACKGROUND: Patients with advanced hepatocellular carcinoma (HCC) and poor liver function lack effective systemic therapies. Low-energy electromagnetic fields (EMFs) can influence cell biological processes via non-thermal effects and may represent a new treatment option. METHODS: This single-site feasibility trial enrolled patients with advanced HCC, Child-Pugh A and B, Eastern Cooperative Oncology Group 0-2. Patients underwent 90-min amplitude-modulated EMF exposure procedures every 2-4 weeks, using the AutEMdev (Autem Therapeutics). Patients could also receive standard care. The primary endpoints were safety and the identification of hemodynamic variability patterns. Exploratory endpoints included health-related quality of life (HRQoL), overall survival (OS). and objective response rate (ORR) using RECIST v1.1. RESULTS: Sixty-six patients with advanced HCC received 539 AutEMdev procedures (median follow-up, 30 months). No serious adverse events occurred during procedures. Self-limiting grade 1 somnolence occurred in 78.7% of patients. Hemodynamic variability during EMF exposure was associated with specific amplitude-modulation frequencies. HRQoL was maintained or improved among patients remaining on treatment. Median OS was 11.3 months (95% confidence interval [CI]: 6.0, 16.6) overall (16.0 months [95% CI: 4.4, 27.6] and 12.0 months [6.4, 17.6] for combination therapy and monotherapy, respectively). ORR was 24.3% (32% and 17% for combination therapy and monotherapy, respectively). CONCLUSION: AutEMdev EMF exposure has an excellent safety profile in patients with advanced HCC. Hemodynamic alterations at personalized frequencies may represent a surrogate of anti-tumor efficacy. NCT01686412.

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Background: Low-frequency electrical stimulation (LFES) is an adjuvant method for heart failure (HF) patients with restrictions to start an exercise. However, the impact on molecular changes in circulating is unknown. We investigated the effects of 10 weeks of home-based LFES on plasma cytokines profile, redox biomarkers, metalloproteinases (MMPs) activity, and exercise performance in HF patients. Methods: Twenty-four HF patients (52.45 ± 9.15 years) with reduced ejection fraction (HFrEF) (EF < 40%), were randomly assigned to a home-based LFES or sham protocol. Plasma cytokines profile was assessed through interleukins, interferon-gamma, and tumor necrosis factor levels. Oxidative stress was evaluated through ferric reducing antioxidant power, thiobarbituric acid-reactive substances, and inducible nitric oxide synthase. The MMPs activity were analyzed by zymography. Cardiorespiratory capacity and muscle strength were evaluated by cardiopulmonary test and isokinetic. Results: LFES was able to increase the active-MMP2 activity post compared to pre-training (0.057 to 0.163, p = 0.0001), while it decreased the active-MMP9 (0.135 to 0.093, p = 0.02). However, it did not elicit changes in cytokines, redox biomarkers, or exercise performance (p > 0.05). Conclusion: LFES protocol is a promising intervention to modulate MMPs activity in HFrEF patients, although with limited functional effects. These preliminary responses may help the muscle to adapt to future mechanical demands dynamically.

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Recently, the sleep-wake states have been analysed using novel complexity measures, complementing the classical analysis of EEGs by frequency bands. This new approach consistently shows a decrease in EEG's complexity during slow-wave sleep, yet it is unclear how cortical oscillations shape these complexity variations. In this work, we analyse how the frequency content of brain signals affects the complexity estimates in freely moving rats. We find that the low-frequency spectrum - including the Delta, Theta, and Sigma frequency bands - drives the complexity changes during the sleep-wake states. This happens because low-frequency oscillations emerge from neuronal population patterns, as we show by recovering the complexity variations during the sleep-wake cycle from micro, meso, and macroscopic recordings. Moreover, we find that the lower frequencies reveal synchronisation patterns across the neocortex, such as a sensory-motor decoupling that happens during REM sleep. Overall, our works shows that EEG's low frequencies are critical in shaping the sleep-wake states' complexity across cortical scales.

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Background: How the prefrontal cortex (PFC) recovers its functionality following lesions remains a conundrum. Recent work has uncovered the importance of transient low-frequency oscillatory activity (LFO; < 4 Hz) for the recovery of an injured brain. We aimed to determine whether persistent cortical oscillatory dynamics contribute to brain capability to support 'normal life' following injury. Methods: In this 9-year prospective longitudinal study (08/2012-2021), we collected data from the patient E.L., a modern-day Phineas Gage, who suffered from lesions, impacting 11% of his total brain mass, to his right PFC and supplementary motor area after his skull was transfixed by an iron rod. A systematic evaluation of clinical, electrophysiologic, brain imaging, neuropsychological and behavioural testing were used to clarify the clinical significance of relationship between LFO discharge and executive dysfunctions and compare E.L.´s disorders to that attributed to Gage (1848), a landmark in the history of neurology and neuroscience. Findings: Selective recruitment of the non-injured left hemisphere during execution of unimanual right-hand movements resulted in the emergence of robust LFO, an EEG-detected marker for disconnection of brain areas, in the damaged right hemisphere. In contrast, recruitment of the damaged right hemisphere during contralateral hand movement, resulted in the co-activation of the left hemisphere and decreased right hemisphere LFO to levels of controls enabling performance, suggesting a target for neuromodulation. Similarly, transcranial magnetic stimulation (TMS), used to create a temporary virtual-lesion over E.L.'s healthy hemisphere, disrupted the modulation of contralateral LFO, disturbing behaviour and impairing executive function tasks. In contrast to Gage, reasoning, planning, working memory, social, sexual and family behaviours eluded clinical inspection by decreasing LFO in the delta frequency range during motor and executive functioning. Interpretation: Our study suggests that modulation of LFO dynamics is an important mechanism by which PFC accommodates neurological injuries, supporting the reports of Gage´s recovery, and represents an attractive target for therapeutic interventions. Funding: Fundação de Amparo Pesquisa Rio de Janeiro (FAPERJ), Universidade Federal do Rio de Janeiro (intramural), and Fiocruz/Ministery of Health (INOVA Fiocruz).

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While theophylline has been extensively studied with multiple polymorphs discovered, there is still currently no conclusive structure for the metastable theophylline form III. In this present work, by combining more widely used techniques such as X-ray diffraction and thermogravimetric analysis with more emerging techniques like low-frequency Raman and terahertz time-domain spectroscopy, to analyze the structure and dynamics of a crystalline system, it was possible to provide further evidence that the form III structure has a theophylline monohydrate structure with the water molecules removed. Solid-state density functional theory simulations were paramount in proving that this proposed structure is correct and explain how vibrational modes within the crystal structures feature and govern polymorphic transitions and the metastable form III. Through the insight provided by both simulated and experimental results, it was possible to decisively conclude the elusive crystal structure of theophylline form III. It was also shown that the correct space group for theophylline monohydrate is not P21/n but, in fact, Pc.

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Low frequency noises are predominant in neonatal intensive care units (NICUs). Some studies affirm that neonates can perceive noises from 113 Hz, and can therefore be affected by sound sources with high spectral content at low frequencies (e.g., incubator engine, air fan). Other studies suggest that reverberation amplifies noise within incubators. In this paper, the reverberation time (T, T 30) within an incubator with standard dimensions was measured in one-third octave bands. To get reliable results, the T was measured in 15 positions at the neonate's ear height, in a room with low T values (to reduce the influence of the room in the results), using an impulsive sound method. Results show a heterogeneous T distribution at the neonate's ear height, with maximum average T differences between positions of 1.07 s. The highest average T of all microphone positions is 2.27 s at 125 Hz, an extremely high mean value for such a small space. As the frequency of electrical devices in America is 60 Hz, some harmonics lay within the one-third octave band of 125 Hz, and therefore may create a very reverberant and inappropriate acoustic environment within the audible spectrum of neonates. As the acoustic environment of the incubator and the room are coupled, it is expected that the results are higher in the NICUs than in the room where the measurements were conducted, as NICUs are more reverberant. Therefore, it is recommended that the T will be limited in the international standards, and that incubator designers take it into account.

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Resumen Objetivo: Caracterizar las reacciones adversas medicamentosas de baja frecuencia de aparición, recibidas en la Unidad Coordinadora Provincial de Farmacovigilancia, del 2014 al 2018. Materiales y Método: Estudio observacional, descriptivo, transversal, de serie de casos en Farmacovigilancia, a partir de la notificación espontánea de sospechas de reacciones adversas en el quinquenio 2014 - 2018, en Matanzas. La información se obtuvo de la base de datos provincial de farmacovigilancia. Se describen las reacciones adversas de baja frecuencia en cuanto a sexo, edad, tipo de efecto adverso, intensidad e imputabilidad. Resultados: Se evaluaron un total de 3,407 reacciones adversas por medicamentos de baja frecuencia. Prevalecieron las clasificadas como ocasionales con 1,741 notificaciones (51.1%). El sexo femenino y los adultos fueron los más reportados. Se afectó en mayor cuantía la piel y anejos; los grupos farmacológicos más reportados fueron los analgésicos, seguido de los antimicrobianos, siendo los fármacos con mayor implicación la dipirona (7.1%) y el ibuprofeno (5.8 %). Predominaron las reacciones adversas por medicamentos de baja frecuencia moderadas (93.3%) y las probables (72.2%) según imputabilidad. Conclusiones: En Matanzas, en el período 2014-2018 se reportaron reacciones adversas a medicamentos de baja frecuencia en su mayoría ocasionales y probables en piel y anejos, producida por analgésicos y antimicrobianos en el sexo femenino y adultos, de moderada intensidad.

Abstract Objective. To characterize the adverse drug reactions of low frequency of appearance, received in the Provincial Coordinating Unit of Pharmacovigilance from 2014 to 2018. Method: Observational, descriptive, cross-sectional study of a series of cases in Pharmacovigilance, based on the spontaneous notification of suspected adverse reactions from 2014 to 2018, in Matanzas. The necessary information was obtained from the pharmacovigilance provincial database. Low-frequency adverse reactions are described in terms of sex, age, type of adverse effect, intensity and imputability, among other variables. Results: There were evaluated a total of 3407 adverse reactions due to low frequency medications. Those classified as occasional prevailed with 1,741 notifications (51.1%). Female sex and adults were the most reported. Analgesics followed by antimicrobials, and the drugs with the greatest involvement were dipyrone (7.1%) and ibuprofen (5.8%). Adverse reactions to moderate low frequency drugs (93.3%) and probable (72.2%) prevailed according to imputability. Conclusions: adverse reactions to low-frequency medications were reported in Matanzas, in the period 2014-2018, mostly occasional and probable in skin and appendages, produced by analgesics and antimicrobials in females and adults, of moderate intensity.

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The zinc transporter 8 (ZnT8) plays an essential role in zinc homeostasis inside pancreatic ß cells, its function is related to the stabilization of insulin hexameric form. Genome-wide association studies (GWAS) have established a positive and negative relationship of ZnT8 variants with type 2 diabetes mellitus (T2DM), exposing a dual and controversial role. The first hypotheses about its role in T2DM indicated a higher risk of developing T2DM for loss of function; nevertheless, recent GWAS of ZnT8 loss-of-function mutations in humans have shown protection against T2DM. With regard to the ZnT8 role in T2DM, most studies have focused on rodent models and common high-risk variants; however, considerable differences between human and rodent models have been found and the new approaches have included lower-frequency variants as a tool to clarify gene functions, allowing a better understanding of the disease and offering possible therapeutic targets. Therefore, this review will discuss the physiological effects of the ZnT8 variants associated with a major and lower risk of T2DM, emphasizing the low- and rare-frequency variants.

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This study proposes an update for the jabuticaba processing chain to obtain valuable coproducts from jabuticaba peels. High-intensity ultrasound (HIUS) technology was evaluated as a more efficient extraction process to obtain two high added-value coproducts: pectin and an anthocyanins-rich extract. The HIUS-assisted extraction of bioactive compounds like anthocyanins from the jabuticaba peels was evaluated. The effects of ultrasound intensity (1.1, 3.7, 7.3, and 13.0 W/cm2) and solvent composition concerning water/ethanol ratio (0, 25, 50, 75, and 100 g water/100 g) were examined. One-step HIUS processing promoted the best recovery of bioactive compounds at an ultrasound intensity of 3.7 W/cm2 and 50 g water/100 g, thus proofing the interaction between ultrasound intensity and the solvent composition has a strong influence on the extraction efficiency of the groups of compounds studied and in the jabuticaba peel antioxidant potential. The confocal laser scanning microscopy confirmed bioactive compounds' exhaustion in the dried jabuticaba peel after the HIUS processing, proving its best recovery. The jabuticaba peel extract exhibited an intense reddish color typical of anthocyanin-rich products at acid pH (4.5). The HIUS technology turned out a promising way to recover these valuable phenolic compounds as a quick, relatively inexpensive, and simple technology that improves the yields and decreases the costs and environmental impacts compared to conventional extraction processes.

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The neonatal intensive care unit (NICU) is a very noisy place as compared to the intrauterine environment. To protect the neonate's health, international guidelines suggest avoiding noise levels above 45 dB in NICUs, but this recommendation is not normally met. The incubator acoustic isolation and the acoustic features of the NICU play important roles in determining the noise measured inside the incubator. In this study, the influence of two types of rooms, one with sound-absorbent covering and the other with reverberant surfaces, on the acoustic isolation of a neonatal incubator was evaluated using three acoustic isolation indexes: the level difference, the apparent sound reduction index, and the standardized level difference. Results show that the acoustic isolation of the incubator is very poor, with a level difference below 11 dBA at all frequencies. At 62.5 Hz, the level difference measured in both rooms exhibits a negative value, indicating that the incubator amplifies the noise coming from the NICU. Isolation of the incubator is poor, and the reverberation time (RT) of the containing room influences RT of the incubator, which is consequently higher when the containing room is reverberant; for example, the incubator RT in the reverberant NICU is 0.72 s higher at 500 Hz than that in a room with sound-absorbent covering.

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Parkinson's disease (PD), the second most frequent neurodegenerative disease, affects significantly life quality by a combination of motor and cognitive disturbances. Although it is traditionally associated with basal ganglia dysfunction, cortical alterations are also involved in disease symptoms. Our objective is to evaluate the alterations in brain dynamics in de novo and recently treated PD subjects using a nonlinear method known as Active Information Storage. In the current research, Active Information Storage (AIS) was used to study the complex dynamics in motor cortex spontaneous activity captured using resting state functional Magnetic Resonance Imaging (rs-fMRI) at early-stage in non-medicated and recently medicated PD subjects. Supplementary to AIS, the fractional Amplitude of Low Frequency Fluctuation (fALFF), which is a better-established technique of analysis of rs-fMRI signals, was also evaluated. Compared to healthy subjects, the AIS values were significantly reduced in PD patients over the analyzed motor cortex regions; differences were also found at less extent using the fALFF measure. Correlations between AIS and fALFF values showed that the measures seem to capture similar neuronal phenomena in rs-fMRI data. The highest sensitivity when detecting group differences revealed by AIS, and not captured by traditional linear approaches, suggests that this measure is a promising tool for the analysis of rs-fMRI neural data in PD.

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The planning process consists of pre-determining an ordered series of actions to accomplish a goal. Previous research showed that the left prefrontal cortex (PFC) is likely to create the strategy for a plan, while the right PFC could be relevant for its update. These roles for the two PFCs need to be ascertained for visuospatial planning, whether communalities or differences exist with other planning tasks. Moreover, the contribution of the posterior parietal cortex (PPC) to planning still lacks evidence. Online repetitive transcranial magnetic stimulation (1 Hz) was used, and 32 participants were involved in the visuospatial planning task in a within-subject design to inhibit either the frontal or the parietal cortex of either the left or the right hemisphere. The goal consisted of evaluating the contribution of these cortical regions, also controlling for gender, in a computerized version of the travelling salesman problem (TSP), the "Maps" task. The results showed that all the stimulated sites produced significant differences in their involvement, reflected in several parameters (such as initial planning and execution times, strategies and heuristics used), with respect to the control group. The roles for the two PFCs were generally confirmed in all measures except path length, while the contribution of the PPC emerged throughout the measures related to the ongoing execution. We concluded that the results obtained with the TSP paradigm were consistent with results obtained using other tasks used to study the planning process (such as the Tower of London) for the evaluation of PFC contribution. In addition, we showed that the contribution of the PPC to the planning process has probably been underestimated.

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Resting-state functional magnetic resonance imaging has been playing an important role in the study of amyotrophic lateral sclerosis (ALS). Although functional connectivity is widely studied, the patterns of spontaneous neural activity of the resting brain are important mechanisms that have been used recently to study a variety of conditions but remain less explored in ALS. Here we have used fractional amplitude of low-frequency fluctuation (fALFF) and regional homogeneity (ReHo) to study the regional dynamics of the resting brain of nondemented ALS patients compared with healthy controls. As expected, we found the sensorimotor network with changes in fALFF and ReHo, and also found the default mode network (DMN), frontoparietal network (FPN), and salience network (SN) altered and the cerebellum, although no structural changes between ALS patients and controls were reported in the regions with fALFF and ReHo changes. We show an altered pattern in the spontaneous low-frequency oscillations that is not confined to the motor areas and reveal a more widespread involvement of nonmotor regions, including those responsible for cognition.

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RATIONALE: The use of electrical stimulation therapy to treat epilepsy is currently being studied in experimental animals and patients. Our study was designed to evaluate the effects of electrical stimulation applied in the thalamic reticular nucleus (TRN) on the development of pentylentetrazole-induced seizures. MATERIALS AND METHODS: Experiments were performed using male Wistar rats with electrodes stereotaxically implanted in the left TRN. Epidural EEG recording screws were implanted in the motor cortex for EEG recording. The rats were classified in seven groups: one sham group, four groups receiving either high- or low-frequency preemptive stimulation for either 10 or 60 minutes, and two groups receiving either high- or low-frequency responsive stimulation for ten minutes. All animals received a single dose of pentylentetrazole throughout five days. EEG recordings were obtained from the cortex and were evaluated to assess ictal behavior more than 45 to 90 minutes. RESULTS: Ten minutes of preemptive high-frequency stimulation in the TRN induced a significant decrease in seizure severity compared to 60 minutes of preemptive stimulation and ten minutes of responsive stimulation. Additionally, ten minutes of preemptive high-frequency stimulation protected against death as aftereffect of status epilepticus. The spike-wave complex frequency was not modified. CONCLUSIONS: These data could contribute to the characterization of the TRN in mediating the initiation and spreading of seizure activity and provide preclinical support for optimal parameters to use to obtain beneficial effects against convulsive activity.

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The induction of both long-term potentiation (LTP) and long-term depression (LTD) of synaptic transmission entails pre- and postsynaptic Ca2+ signals, which represent transient increments in cytoplasmic free Ca2+ concentration. In diverse synapse types, Ca2+ release from intracellular stores contributes to amplify the Ca2+ signals initially generated by activation of neuronal Ca2+ entry pathways. Here, we used hippocampal slices from young male rats to evaluate whether pharmacological activation or inhibition of Ca2+ release from the endoplasmic reticulum (ER) mediated by ryanodine receptor (RyR) channels modifies LTD induction at Schaffer collateral-CA1 synapses. Pre-incubation of slices with ryanodine (1 µM, 1 h) or caffeine (1 mM, 30 min) to promote RyR-mediated Ca2+ release facilitated LTD induction by low frequency stimulation (LFS), but did not affect the amplitude of synaptic transmission, the profiles of field excitatory postsynaptic potentials (fEPSP) or the paired-pulse (PP) responses. Conversely, treatment with inhibitory ryanodine (20 µM, 1 h) to suppress RyR-mediated Ca2+ release prevented LTD induction, but did not affect baseline synaptic transmission or PP responses. Previous literature reports indicate that LTD induction requires presynaptic CaMKII activity. We found that 1 h after applying the LTD induction protocol, slices displayed a significant increase in CaMKII phosphorylation relative to the levels exhibited by un-stimulated (naïve) slices. In addition, LTD induction (1 h) enhanced the phosphorylation of the presynaptic protein Synapsin I at a CaMKII-dependent phosphorylation site, indicating that LTD induction stimulates presynaptic CaMKII activity. Pre-incubation of slices with 20 µM ryanodine abolished the increased CaMKII and Synapsin I phosphorylation induced by LTD, whereas naïve slices pre-incubated with inhibitory ryanodine displayed similar CaMKII and Synapsin I phosphorylation levels as naïve control slices. We posit that inhibitory ryanodine suppressed LTD-induced presynaptic CaMKII activity, as evidenced by the suppression of Synapsin I phosphorylation induced by LTD. Accordingly, we propose that presynaptic RyR-mediated Ca2+ signals contribute to LTD induction at Schaffer collateral-CA1 synapses.

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In this work, an ultrasound experimental setup was designed to investigate the feasibility of using low-frequency ultrasonic waves as a substitute to reduce the consumption of chemical demulsifiers in the pretreatment of crude oil. The experiments were planned to study the effects of irradiation time, ultrasonic field intensity and initial water content on the efficiency of separation. The results of experiments showed that by selecting a proper irradiation time and field intensity, it is possible to decrease the usage of demulsifiers by 50%. Moreover, a population balance model was proposed to explicate the experimental data. A hybrid coalescence model was developed to determine the frequency of aggregation. The parameters of the model were estimated by linear regression. The parameter estimation was performed using a parallel execution of the particle swarm optimization algorithm. The results of the model showed a decent agreement with the experimental data.