Northern Ohio Recovery Association

Histone modifications play essential roles in regulating chromatin accessibility and downstream transcription, serving as critical determinants of cell identity and function. However, the diversity of histone post-translational modifications (PTMs) and their tendency to be studied in isolation limits our understanding of their coordinated roles in shaping cellular states. Conventional flow cytometry methods for histone PTM assessment suffer from low multiplexing capacity and typically require nuclear isolation, resulting in significant sample loss and an incomplete picture of the overall cell state. Here, we present EpiFlow, a spectral flow cytometry protocol designed for multiparametric analysis of histone PTMs within whole cells. By utilizing a 96-well plate format and preserving the cell entirely, EpiFlow improves throughput and efficiency while retaining sample integrity. This method resolves subtle variations in histone PTMs within neural progenitor cells, capturing distinct chromatin states across the cell cycle and correlating them with several markers. Furthermore, we provide an open-access GitHub repository containing detailed protocols and analysis workflows, ensuring reproducibility and accessibility of this approach. EpiFlow offers a robust framework for exploring chromatin dynamics, with broad implications for advancing fundamental research and therapeutic research.

ABSTRACTCortical spreading depression (CSD) is a wave of pronounced depolarization of brain tissue accompanied by substantial shifts in ionic concentrations and cellular swelling. Here, we validate a computational framework for modelling electrical potentials, ionic movement, and cellular swelling in brain tissue during CSD. We consider different model variations representing wild type or knock-out/knock-down mice and systematically compare the numerical results with reports from a selection of experimental studies. We find that the data for several CSD hallmarks obtained computationally, including wave propagation speed, direct current shift duration, peak in extracellular K+ concentration as well as a pronounced shrinkage of extracellular space, are well in line with what has previously been observed experimentally. Further, we assess how key model parameters including cellular diffusivity, structural ratios, membrane water and/or K+ permeabilities affect the set of CSD characteristics.Significance StatementMovement of ions and molecules in and between cellular compartments is fundamental for brain function. Cortical spreading depression (CSD) is associated with dramatic failure of brain ion homeostasis. Better understanding the sequence of events in CSD could thus provide new insight into physiological processes in the brain. Despite extensive experimental research over the last decades, even basic questions related to mechanisms underlying CSD remain unanswered. Computational modelling can play an important role going forward, since simulation studies can address hypotheses that are difficult to target experimentally. Here, we assess the physiological validity of a novel mathematical framework for detailed modelling of brain electrodiffusion and osmosis – and provide a platform for in silico studies of CSD and other cerebral electro-mechanical phenomena.

Introduction: Literature on the effectiveness of simulation based medical education programs used in acute ischemic stroke (AIS) care is scant. In an effort to improve coordination and door to needle time (DNT) for AIS care, a stroke simulation education training program for neurology nursing staff and neurology residents was implemented in a comprehensive stroke center. Methods: Hospital stroke registry was used for retrospective analysis. The study population was defined as all patients treated with IV-tPA for AIS in the emergency room from October 2008 to September 2014. Simulation training was implemented yearly, for a three month period starting from July 2011. All neurology residents and a group of nurses trained to respond to all AIS cases participated. Simulations were standardized, using deliberate practice with a trained live actor portraying stroke vignettes in the presence of a board certified vascular neurologist. During the period of study, there were no changes in Emergency Department stroke triage protocol, or changes in first provider response to AIS. The data was analyzed using IBM SPSS24 software. Results: We identified 448 patients admitted with AIS who were treated with IV-tPA. The average DNT on univariate analysis before and after intervention was 67.9 and 58.3 minutes [p <0.001]. A multivariate linear regression analysis was performed controlling for age, night/day shift, weekday/weekend, and blood pressure at presentation (>185/110). After controlling for confounders we found that simulation training independently reduced the DNT by 9.64 minutes [95% confidence interval (CI) 4.01 - 15.28, p=0.001]. Amongst other co-variates, only the systolic blood pressure >185 was associated with 14.27 minutes of delay in DNT [95% CI 3.36 - 25.191, p=0.011]. Conclusion: Time to thrombolysis from symptom onset is a critical factor in AIS management and evidence shows improving the DNT could improve patient outcomes. In our six year study, integration of simulation based medical education for AIS reduced the average DNT by 9.64 minutes in multivariate analysis. Simulation based medical education therefore should be considered as a standard process for providers involved in the care of AIS patients receiving thrombolytic treatment.

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The risk of recurrence of a congenital cardiovascular malformation in a child having one parent with congenital heart disease has been determined for each of the seven most common anomalies presently compatible with survival to reproductive age. The range of risk is 2.5% to 4.3% depending on the lesion. This is within the range of expectation for the model of multifactorial inheritance previously used to predict recurrence in other first-degree relatives of probands (siblings and parents) with congenital heart disease. The cardiovascular abnormality occurring in the child was most often the same as in the parent or was a closely related variant of it.