Radiation Exposure in the Acute Phase after Aneurysmal Subarachnoid Hemorrhage in the Era of CT Perfusion.

PURPOSE: Aneurysmal subarachnoid hemorrhage (aSAH) is associated with a high risk of developing multiple complications requiring further diagnostics including imaging associated with radiation exposure (RE). Since aSAH often affects younger patients, the obtained cumulative RE may have serious long-term health consequences. The aim of this study was to calculate the cumulative RE in the acute phase after aSAH and to identify contributors to RE. Additionally, we investigated whether there is a correlation of RE with outcome. METHODS: A retrospective analysis of patients with aSAH treated at our department from 2012 to 2018 was performed. The radiation dose of every single cranial radiological examination was calculated for every patient. The outcome was assessed according to the modified Rankin scale (mRS) 3 months after ictus. Factors associated with high RE were evaluated and the correlation of RE with outcome was assessed. RESULTS: In 268 included consecutive patients, the mean cumulative RE per patient was 39.95 mSv, ranging from 2 to 265.5 mSv. A higher RE correlated with delayed cerebral ischemia (r = 0.52, p < 0.0001), delayed infarction (r = 0.25, p < 0.0001), delayed ischemic neurological deficits (r = 0.29, p < 0.0001) and transcranial Doppler (TCD)-vasospasm (r = 0.34, p < 0.0001). Independent predictors of outcome were age (p = 0.0001), World Federation of Neurosurgical Societies (WFNS) grade (p < 0.0001) and delayed infarction (p = 0.0004), while RE did not correlate with outcome. CONCLUSION: There is a considerable imaging-related RE in aSAH patients. A meticulous decision-making process and imaging protocols with lower RE for the deployment of CT-based and fluoroscopy-based imaging is indicated in order to minimize the risk for radiation-mediated heath consequences in this patient population.

An intact keratin network is crucial for mechanical integrity and barrier function in keratinocyte cell sheets.

The isotype-specific composition of the keratin cytoskeleton is important for strong adhesion, force resilience, and barrier function of the epidermis. However, the mechanisms by which keratins regulate these functions are still incompletely understood. In this study, the role and significance of the keratin network for mechanical integrity, force transmission, and barrier formation were analyzed in murine keratinocytes. Following the time-course of single-cell wound closure, wild-type (WT) cells slowly closed the gap in a collective fashion involving tightly connected neighboring cells. In contrast, the mechanical response of neighboring cells was compromised in keratin-deficient cells, causing an increased wound area initially and an inefficient overall wound closure. Furthermore, the loss of the keratin network led to impaired, fragmented cell-cell junctions, and triggered a profound change in the overall cellular actomyosin architecture. Electric cell-substrate impedance sensing of cell junctions revealed a dysfunctional barrier in knockout (Kty-/-) cells compared to WT cells. These findings demonstrate that Kty-/- cells display a novel phenotype characterized by loss of mechanocoupling and failure to form a functional barrier. Re-expression of K5/K14 rescued the barrier defect to a significant extent and reestablished the mechanocoupling with remaining discrepancies likely due to the low abundance of keratins in that setting. Our study reveals the major role of the keratin network for mechanical homeostasis and barrier functionality in keratinocyte layers.

Stiffness of MDCK II Cells Depends on Confluency and Cell Size.

Mechanical phenotyping of adherent cells has become a serious tool in cell biology to understand how cells respond to their environment and eventually to identify disease patterns such as the malignancy of cancer cells. In the steady state, homeostasis is of pivotal importance, and cells strive to maintain their internal stresses even in challenging environments and in response to external chemical and mechanical stimuli. However, a major problem exists in determining mechanical properties because many techniques, such as atomic force microscopy, that assess these properties of adherent cells locally can only address a limited number of cells and provide elastic moduli that vary substantially from cell to cell. The origin of this spread in stiffness values is largely unknown and might limit the significance of measurements. Possible reasons for the disparity are variations in cell shape and size, as well as biological reasons such as the cell cycle or polarization state of the cell. Here, we show that stiffness of adherent epithelial cells rises with increasing projected apical cell area in a nonlinear fashion. This size stiffening not only occurs as a consequence of varying cell-seeding densities, it can also be observed within a small area of a particular cell culture. Experiments with single adherent cells attached to defined areas via microcontact printing show that size stiffening is limited to cells of a confluent monolayer. This leads to the conclusion that cells possibly regulate their size distribution through cortical stress, which is enhanced in larger cells and reduced in smaller cells.

Single-Cell Defects Cause a Long-Range Mechanical Response in a Confluent Epithelial Cell Layer.

Epithelial cells are responsible for tissue homeostasis and form a barrier to maintain chemical gradients and mechanical integrity. Therefore, rapid wound closure is crucial for proper tissue function and restoring homeostasis. In this study, the mechanical properties of cells surrounding a single-cell wound are investigated during closure of the defect. The single-cell wound is induced in an intact layer using micropipette action and responses in neighboring cells are monitored with atomic force microscopy. Direct neighbors reveal a rise in the apparent pretension, which is dominated by cortical tension. The same effect was observed for a single-cell wound induced by laser ablation and during closure of a not fully confluent layer. Moreover, changes in the apparent pretension are far reaching and persist even in cells separated by three cell widths from the defect. This shows that epithelial cells respond to minimal wounds in a collective fashion by increased contractility with substantial reach.