Membrane potential (or voltage) imaging in acute brain slices is widely used in fundamental and applied neuroscience even in an age characterised by the surge of in-vivo recording. This optical method remarkably increased its ability in providing key information both at the level of single cell and at the level of populations and networks. But, more importantly, technical progresses occurred in the last two decades enlarged the potential pool of voltage imaging users from relatively few specialists to scientists in many fields including those working in industry. This review aims at offering an updated guideline for anyone wishing to perform voltage imaging in brain slices. We analyse the technical and practical aspects to consider in the choice between using an organic voltage sensitive dye (VSD) or a genetically encoded voltage indicator (GEVI). We then examine advantages and disadvantages of using the straightforward approach of wide-field illumination, versus other methods providing higher spatial resolution and optical sectioning. Other choices are analysed in particular for users adopting VSDs who are confronted with the decision of recording from single neurons or from neuronal populations. The review is concluded by exploring the prospective of applications in the near future.

Endovascular Coarctoplasty (CoAP) with stenting is the treatment of choice in Coarctation of Aorta (CoA). Literature on the harms or benefits of CoAP in Eisenmenger's syndrome (ES) is scarce. There is the hypothetical possibility of an increase in right-to-left intracardiac shunting once the coarctation is relieved. We report a 17-year-old male adolescent who presented with secondary hypertension, cyanosis, and lower limb claudication. He was diagnosed as a case of ventricular septal defect (VSD), patent ductus arteriosus (PDA), and CoA with associated ES. He underwent successful balloon CoAP with no clinical or invasive hemodynamic deterioration. At one-year follow-up, there was no clinical or echocardiographic evidence of complications from ES or restenosis. This case highlights that CoA with ES can be safely managed by CoAP with good long-term results. The proposed pathophysiology for the uneventful procedure is an extension of our understanding of the 'shunt dynamics' in cyanotic spells in patients with congenital cyanotic heart diseases.

Exposure to environmental chemicals such as bisphenol A (BPA) may affect neural circuit function in subtle and spatially distributed ways that are difficult to attribute to a single circuit element. Here, we developed a voltage-sensitive dye (VSD) imaging analysis pipeline for acute male mouse hippocampal slices. This approach measures stimulus-evoked fractional changes in VSD fluorescence (ΔF/F 0) to generate pixel-wise normalized maps, averages response magnitude within anatomically defined regions of interest (ROIs), and visualizes control-versus-compound differences using pixel-wise statistical maps before and after false discovery rate (FDR) adjustment. We applied this pipeline to compare the acute effects of BPA with those of two structurally related phenolic analogs. Evoked responses were quantified by their peak amplitude and time integral across hippocampal trisynaptic pathways at two stimulation levels determined from input-output relationships, with or without the GABAA receptor antagonist gabazine (SR-95531; 10 μM). The phenolic analogs reduced circuit responses across multiple ROI-defined conditions, whereas BPA produced comparatively limited effects. These alterations varied with pathway, subfield, age, response metric, and inhibitory state. These findings demonstrate the utility of integrating ROI averages with pixel-wise statistical maps for detecting condition-dependent circuit-level differences between structurally related compounds and indicate that BPA-related phenolic analogs can differ from BPA itself in their acute effects on hippocampal circuitry.Significance Statement Exposure to environmental chemicals may alter brain circuit activity in subtle, spatially distributed ways. We developed a voltage-sensitive dye imaging pipeline for acute mouse hippocampal slices that combines regional summaries with pixel-wise statistical maps, quantifying regional response magnitude while localizing where statistically supported differences are expressed within evoked response fields. Using this framework, we compared the acute hippocampal circuit effects of bisphenol A (BPA) with those of two structurally related phenolic analogs. Under acute slice conditions, statistically supported response reductions were detected more consistently for the phenolic analogs than for BPA, depending on age, inhibitory state, hippocampal subfield, and response metric. These results show how integrated regional and pixel-wise analysis improves circuit-level phenotyping of structurally related compounds.

BackgroundSustained total all-region (STAR) perfusion is an emerging technique that provides continuous perfusion to the brain, lower body, and heart with mild hypothermia. We hypothesized that using STAR perfusion would result in improved postoperative outcomes.MethodsThe 4 cohorts evaluated (total n = 105) included 1) STAR perfusion for aortic arch reconstruction (Arch w/ STAR), 2) STAR perfusion for aortic arch reconstruction with subsequent cardiac arrest for ventricular septal defect/atrial septal defect (VSD/ASD) (Arch/ASD/VSD w/ STAR), 3) hypothermic circulatory arrest (HCA) with cerebral perfusion (CP) for aortic arch reconstruction (Arch w/ HCA/CP), and 4) hypothermic circulatory arrest with cerebral perfusion for aortic arch reconstruction and VSD/ASD closure (Arch/ASD/VSD w/ HCA/CP). Primary outcomes were 1-year mortality and postoperative kidney, neurologic, and cardiac dysfunction.ResultsOperative times differed between the cohorts, with Arch w/ STAR patients having the lowest operative times. Peak postoperative lactate was lower in patients who underwent Arch w/ STAR compared with Arch w/ HCA/CP. On multivariable analysis, patients in the Arch w/ HCA/CP demonstrated higher peak postoperative lactate. Postoperative creatinine was primarily predicted by baseline renal function on multivariable analysis. Arch w/ STAR and Arch/ASD/VSD w/ STAR patients had a shorter time to chest closure, and Arch/ASD/VSD w/ STAR patients produced more urine at 24 h postoperative compared with patients who underwent Arch/ASD/VSD w/ HCA/CP. There were no differences in neurologic outcomes. No mortality occurred in our STAR cohorts.ConclusionSTAR perfusion is an emerging technique that is associated with lower peak lactate levels and shorter operative times compared with patients undergoing traditional methods of aortic reconstruction.

Voltage-gated sodium (NaV) channels are critical transmembrane proteins responsible for the initiation and propagation of electrical signals in excitable tissues. They consist of a large α subunit and auxiliary β subunits. Together, the voltage sensing, ion selectivity, and rapid gating transitions regulate neuronal, muscular, and cardiac excitability. Recent cryo-electron microscopy breakthroughs have transformed the understanding of NaV structure-function relationships, revealing the asymmetric organization of voltage-sensing domains (VSD), the architecture of the DEKA selectivity filter, the mechanical coupling between S4-S5 linkers and S6 gating helices, and the allosteric mechanism by which the IFMT motif mediates fast inactivation. Across the nine human NaV isoforms, subtle structural divergences underlie distinct tissue distributions, biophysical properties, and disease susceptibilities, explaining the diverse channelopathies that range from epilepsy and cardiac arrhythmias to skeletal muscle disorders and painful neuropathies. These high-resolution structures also illuminate isoform-specific pharmacological pockets, including membrane-dependent cavities in VSD4 and dynamic fenestrations that govern state-dependent drug access, enabling the rational design of small molecules, peptide toxins, and hybrid therapeutics with improved selectivity. Advances in structural biology, biochemistry, and computation have established an integrated mechanistic model that connects NaV channels architecture to gating, inactivation, isoform-specific function, and human disease. This growing molecular atlas now supports next-generation precision therapies aimed at selectively modulating NaV channels activity in neurological, muscular, cardiac, and pain disorders.

Precise extracellular matrix (ECM) coordination is essential for cardiac septation, yet the molecular etiology of human isolated perimembranous ventricular septal defects (VSD) remains elusive. Here, we constructed a high-resolution single-cell transcriptomic atlas of human VSD, benchmarking the pathological state against a 7-22 week developmental trajectory. Integrating these single-cell profiles with a reference-based spatial transcriptomic map, we deconvolved the distinct molecular signatures of membranous and muscular septal regions. We found that aberrant cardiomyocyte hypertrophy operates alongside a systemic failure in the non-myocyte microenvironment. Crucially, the ECM regulatory network is dysregulated, driven by attenuated endothelial-fibroblast crosstalk and downregulated THBS1-integrin signaling. Supported by in vitro assays, this dysregulation impairs key endothelial-to-mesenchymal transition (EndoMT) programs and transcriptionally suppresses MMP2. Spatial reconstruction confirmed this maturation block compromises both septal regions, indicating a global remodeling arrest that precludes physical closure. Validated by in situ immunofluorescence and a robust correlation between reduced maternal serum MMP2 levels and defect size, our study portrays VSD as a disease driven by microenvironmental insufficiency and arrested remodeling, providing an integrative framework for understanding human cardiac dysmorphogenesis.

Electroencephalography (EEG) has been investigated as a noninvasive approach for characterizing brain activity in neurodegenerative conditions. This study evaluated whether multidomain EEG biomarkers could distinguish Alzheimer's disease (AD), frontotemporal dementia (FTD), and healthy controls (HC). The publicly available dataset contained resting-state, eyes-closed EEG recordings from 88 participants, including 36 with AD, 23 with FTD, and 29 healthy controls. Multidomain EEG biomarkers and demographic features were extracted for machine learning classification. The EEG-derived features included root-mean-square (RMS), power spectral density (PSD), and entropy measures. A Random Forest classifier was trained using the combined feature set and interpreted using explainable artificial intelligence. Feature-stability and Cohen's d effect-size analyses were performed to evaluate feature contributions, followed by an ablation analysis. Entropy-based features showed the lowest contribution to classification. The complete multidomain model achieved 96% training accuracy and 92% test accuracy. After the removal of the entropy features, test accuracy increased to 94%. These results describe the classification performance of the selected multidomain EEG and demographic features within the Random Forest framework.

Eisenmenger syndrome (ES) represents the advanced form of pulmonary arterial hypertension (PAH) associated with a reversed or bidirectional shunt at the atrioventricular or aortopulmonary level. Pregnant women have a higher risk of developing volume overload due to this syndrome, compounded by the physiological changes of pregnancy, which can result in rapidly progressive cardiopulmonary decompensation and thrombotic complications. Due to the substantial risk to both the mother and fetus, women are advised not to conceive or to terminate the pregnancy in the first trimester. However, with appropriate management and careful consideration of all potential risks, positive neonatal and maternal outcomes can be achieved. We report a case of an 18-year-old primigravida with PAH and preeclampsia at 35+5 weeks' period of gestation (POG), who presented to our center for the first time with lower limb and vulval swelling.

Feline calicivirus (FCV) shows extensive genetic and antigenic diversity and typically causes oral-respiratory disease (ORD), although some isolates are associated with virulent systemic disease (VSD). The molecular basis of FCV-associated VSD remains unclear and cannot be attributed to a single VP1 residue, phylogenetic lineage, or recombination event. Here, we comparatively characterized three VSD-associated isolates (XA13_VSD, GZ_VSD, and AH1_VSD) and one ORD-associated isolate (SH2022_ORD) by integrating phylogenetic background, modular genomic variation, in vitro replication, experimental pathogenicity, tissue viral RNA distribution, and serological cross-neutralization. All four isolates belonged to genogroup GI, but the VSD-associated isolates were distributed across different GI sublineages. RDP5 analysis of 253 complete genomes identified 90 recombination events, including 27 spanning the Region A/Region B boundary; AH1_VSD showed a representative modular recombination pattern near the ORF1/ORF2 junction. Within this four-isolate panel, the three VSD-associated isolates showed higher in vitro replication levels and caused more severe disease than SH2022_ORD in experimentally infected cats, accompanied by greater viral RNA shedding, whole-blood FCV RNA detection at 3-9 dpi, broader tissue viral RNA distribution, and more severe histopathological lesions. After homologous neutralizing titers were standardized to 2 [1] [0], AH1_VSD antiserum displayed the broadest cross-neutralizing profile within the tested panel. Collectively, these findings support a multifactorial view of FCV-associated systemic disease involving genomic background, replication and pathogenicity traits, viral dissemination, tissue involvement, and capsid antigenic variation. The comparatively broad cross-neutralization profile of AH1_VSD provides a rationale for its further evaluation as a potential parental candidate for attenuation and vaccine development.