Citations

BACKGROUND: Type 2 long-QT syndrome results from KCNH2 -encoded Kv11.1 loss-of-function, but whether secondary inward-current dysfunction amplifies dysfunction remains unclear. METHODS: We characterized sodium ( SCN5A , I Na ) and L-type calcium channel ( CACNA1C , I Ca ) function in KCNH2-G604S and variant-corrected isogenic control induced pluripotent stem cell–derived cardiomyocytes (iPSC-CMs). Whole-cell patch-clamp measured action potential duration at 90% repolarization (APD90), and potassium (I Kr ), sodium (I Na ), and calcium (I Ca, L ) currents. Fluo-4 imaging quantified calcium transients. A modified Kernik-Clancy iPSC-CM model incorporated experimentally measured persistent I Na,L and I Ca,L fractions and simulated isolated and combined current defects. RESULTS: KCNH2-G604S caused severe I Kr loss-of-function (72.5% reduction). APD90 was prolonged in G604S-iPSC-CMs (765±58 ms) compared with isogenic controls (368±37 ms; P <0.005), with early afterdepolarizations in 50% of G604S cells. Nifedipine, verapamil, and mexiletine successfully shortened APD90 and abolished early afterdepolarizations. Compared with isogenic control, I Na,L (0.60±0.1% versus 0.26±0.06%; P <0.01), persistent I Ca,L (2.88±0.55% versus 1.26±0.35%; P <0.05), and calcium transient decay times ( P <0.0001) were greater in G604S iPSC-CMs. Increased persistent I Ca,L was phenocopied by E-4031 in isogenic controls and reduced by nifedipine in G604S iPSC-CMs. In silico modeling revealed that the I Kr defect alone prolonged APD90 to only 620 ms. The addition of the observed I Na,L defect had a small additive effect (+20 ms); however, incorporating persistent I Ca,L synergistically extended APD90 to 758 ms, replicating the experimental phenotype. Flux dynamics analysis indicated that this persistent Ca 2+ entry suppresses the forward-mode sodium-calcium exchanger, forcing a compensatory reliance on sarcoplasmic-endoplasmic reticulum Ca-ATPase that primes for calcium instability. CONCLUSIONS: The KCNH2-G604S variant drives a severe multihit phenotype via primary I Kr loss-of-function and secondary state-dependent gain-of-function in persistent I Ca,L . These comprehensive validations demonstrate that the remodeled persistent I Ca,L acts as the essential amplifier driving extreme action potential duration prolongation and Ca 2+ mishandling.

TAR DNA-binding protein 43 (TDP-43) pathology is a major contributor to cognitive decline in Alzheimer’s disease (AD), yet the cellular substrates and circuit-level consequences of its limbic accumulation remain poorly defined. Here, we combined multiplex immunofluorescence with 3D Artificial Intelligence (AI)-based object detection and quantitative colocalization to map phosphorylated TDP-43 (pTDP-43) across neuronal subtypes in the amygdala, hippocampal subfields, and entorhinal cortex (ERC) in human AD, AD with Lewy body disease (AD-LBD), and primary age-related tauopathy (PART). Across 15 deeply phenotyped cases, pTDP-43 showed preferential accumulation in neuromodulatory noradrenergic neurons, with more limited involvement of glutamatergic terminals and cholinergic populations. Noradrenergic neurons marked by dopamine β-hydroxylase (DβH) demonstrated extensive colocalization with pTDP-43 across regions, correlating with APOE genotype, amyloid burden, and tau staging in this cross-sectional cohort, suggesting that neuromodulatory circuits might represent a major convergence point for AD-related molecular stressors. Dopaminergic (tyrosine hydroxylase (TH)) and cholinergic (choline acetyltransferase (ChAT)) neurons showed more restricted but pathology-dependent involvement, with LBD consistently showing lower pTDP-43 burden across multiple transmitter systems. Glutamatergic vesicular glutamate transporter-1 (VGLUT1)-positive neurons and terminals exhibited minimal colocalization except in PART. Principal component (PCA) and hierarchical clustering analyses (HCA) integrating pTDP-43 colocalization with neuropathologic staging metrics (A, B, C, Thal, Braak, LBD) reliably separated PART, AD-TDP, and AD-TDP-LBD groups, with LBD emerging as a dominant modifier of cellular vulnerability. Together, these findings point out that TDP-43 pathology in AD preferentially targets neuromodulatory circuits, is strongly shaped by coexisting synucleinopathy, and provides a cell-type-resolved framework for understanding how TDP-43 contributes to limbic network dysfunction in neurodegeneration.

Background Adenoid cystic carcinoma (ACC) is a rare malignancy characterized by indolent growth, frequent perineural invasion, and a propensity for locally advanced disease involving the skull base. High-dose radiation to complex target volumes must be balanced with minimizing exposure to adjacent organs at risk, making particle therapy attractive. Proton beam therapy (PBT) and carbon ion radiotherapy (CIRT) provide favorable dose distributions compared with photon-based radiotherapy, while CIRT offers additional theoretical radiobiologic advantages through its higher linear energy transfer and relative biological effectiveness. However, whether these properties translate into superior clinical outcomes remains uncertain. Methods This perspective reviews physical and radiobiologic characteristics of PBT and CIRT and evaluates contemporary clinical evidence supporting their use in ACC. Available studies demonstrate favorable local control, overall survival, and toxicity outcomes with both modalities but have not established a clear clinical advantage of CIRT over modern PBT. Interpretation is limited by heterogeneous patient populations, treatment techniques, fractionation schedules, institutional practices, follow-up, and the absence of prospective data. Additionally, differences in infrastructure, cost, geographic availability, and reimbursement influence treatment selection and patient access. Results Current evidence supports PBT as an efficacious and broadly applicable treatment option for ACC, particularly for complex skull base and perineural target volumes. Future research should prioritize comparative dosimetry, refinement of biologic dose modeling, multicenter clinical outcomes, contemporary cost-effectiveness analyses, and patient selection. Expanding equitable access to particle therapy should remain a parallel objective. Conclusion Current literature supports PBT in the treatment of ACC. Reported treatment outcomes are comparable to CIRT.

Abstract Background The rapid growth of FDA‐licensed and clinical trial cell and gene therapy products is causing significant growing pains for laboratories onboarding and processing these products for clinical use. While clinical programs have traditional and supplemental reimbursement models, many services provided by the laboratories are not directly reimbursed. Compounding this issue is the increasing diversity and complexity of products with unique processing requirements leading to resource‐intensive workflows. Study Design and Methods Our institution developed a three‐tiered cost structure for services provided by the cellular therapy laboratory billed to industry‐sponsored, externally manufactured clinical trials based on the level of start‐up and per product handling complexity. A list of tasks and time required to complete the tasks was recorded to account for the complexity of the specific trial. This data was used to determine the total cost billed to onboard and carry out cell processing and handling tasks by the cellular therapy laboratory to the principal investigator of the clinical trial. Results Our tiered fee structure enabled a more accurate accounting of laboratory personnel time based on the complexity of the clinical trial products, enabling our laboratory to recoup necessary costs. Conclusion A tiered fee structure corresponding to higher levels of onboarding and administrative complexities can more accurately estimate costs to enable cell processing laboratories to remain financially sustainable and continue to deliver cell and gene therapy products to patients.

Introduction Human cytomegalovirus (CMV) is the leading infectious cause of congenital birth defects worldwide. The placenta is a major route of vertical transmission, and both immune and trophoblast cells are susceptible to CMV infection. This study aimed to spatially characterize gene and protein expression changes associated with CMV infection in the human placenta. Methods In this exploratory study, CMV+ and control placentae were analyzed using the NanoString GeoMx Digital Spatial Profiler to assess compartment-specific transcriptomic and proteomic profiles in trophoblast and immune cell populations. Results CMV+ placentae showed increased immune activation in both trophoblast and immune cells, including pathways related to immune-cell proliferation, differentiation, lymphopoiesis, and T-cell development. Infected placentae also showed altered tissue repair and remodeling pathways, along with changes in proapoptotic proteins and cell death-associated transcripts. Discussion Given the limited sample size, these findings are descriptive and hypothesis-generating. They provide preliminary insight into CMV-associated placental immune dysregulation and identify candidate pathways for investigation in larger, prospective studies.

Background: Cefazolin and antistaphylococcal penicillins (ASP) are first-line agents for methicillin-susceptible Staphylococcus aureus (MSSA) bacteremia. These agents are substantially protein-bound; however, scant data exist evaluating the impact of hypoalbuminemia on microbiologic and patient outcomes for this indication. Objective: The purpose of this study was to assess whether hypoalbuminemia is associated with delayed blood culture clearance in patients with MSSA bacteremia being treated with cefazolin or ASPs as definitive therapy. Methods: This retrospective cohort study included patients admitted for MSSA bacteremia between January 1, 2006 and December 31, 2019, with a serum albumin obtained between the date of an initial positive blood culture result and 72 hours following initiation of definitive therapy with cefazolin or an ASP. The primary outcome was the rate of persistently positive blood cultures, defined as failure to clear blood cultures within 72 hours of definitive therapy initiation. Patients with a serum albumin ≤2.5 g/dL were compared with those with an albumin >2.5 g/dL. Propensity score weighting was used to account for differences in cohort characteristics. Results: A total of 123 patients were included. After propensity score weighting, patients with hypoalbuminemia had a higher rate of persistently positive blood cultures, both in the overall cohort (45.0% vs 13.2%, P < .001) and in the subgroups of cefazolin (33.0% vs 9.1%, P = .015) and ASP (60.1% vs 18.3%, P = .001) when analyzed separately. After propensity weighting in the combined cohort, the length of stay was longer in the hypoalbuminemic cohort, but no differences in mortality were observed. Conclusions and Relevance: Hypoalbuminemia was associated with a delayed 72-hour blood culture clearance in patients with MSSA bacteremia treated with either cefazolin or ASPs.

ABSTRACT Objective To determine whether patients with hypermobile Ehlers‐Danlos syndrome (hEDS) or hypermobility spectrum disorders (HSD) and inducible laryngeal obstruction (ILO) have more voice, cough, swallowing, GI issues, and MC activation than patients without ILO. Methods This retrospective study of 2162 patients meeting 2017 diagnostic criteria for hEDS ( n = 507) or HSD ( n = 1655) who were seen at the EDS Clinic in Mayo Clinic Florida from November 2019 to June 2025 compared patients who screened positive for ILO to those who screened negative. Patients were examined for self‐reported symptoms of voice, cough, swallow, and GI issues as well as MC activation. Results Among patients with hEDS or HSD, 23.1% and 17.1% respectively screened positive for ILO. Patients with ILO had significantly more voice, cough, and swallow issues than patients without ILO (all p ≤ 0.007). Both diagnoses had more gastroesophageal reflux disease (GERD) (hEDS: 55.6% vs. 40.9%, p = 0.004; HSD: 55.8% vs. 42.2%, p < 0.0001). Additionally, both hEDS and HSD patients with ILO self‐reported 18 allergy‐related symptoms more often than those without ILO and had higher MC scores indicating widespread MC activation (all p < 0.0001). Cough, dysphagia, and elevated MC score were statistically significant predictors of ILO in both hEDS and HSD within a logistic regression analysis. Conclusions Chronic cough, dysphagia, and elevated MC symptom burden were the strongest independent predictors of ILO across both hEDS and HSD, underscoring the need for routine multidisciplinary laryngeal assessment in this population. Level of Evidence 3.