The histone deacetylase inhibitor, Trichostatin A, enhances radiation sensitivity and accumulation of gammaH2A.X

Histone deacetylase inhibitors have been shown to induce numerous biologic effects including, altering cell cycle distribution, cytostasis and in certain cases apoptosis. Given their ability to disrupt critical biological processes in cancer cells, these agents are emerging as potential therapeutics for cancer. Recently, it has been identified that histone deacetylase inhibitors can also efficiently enhance the radiation sensitivity of cells, both in vitro and in vivo. In this study, we investigated whether the potent histone deacetylase inhibitor, Trichostatin A, modulates the radiation sensitivity of human erythroleukemic K562 cells. The endpoints we used were clonogenic survival, apoptosis and gammaH2AX immunoprecipitations of soluble chromatin. The findings from clonogenic survival assays indicated that incubation with Trichostatin A 24 hours prior to irradiation enhances the radiation sensitivity of K562 cells. The dose modification factors ranged from 1.1 when cells were incubated with 0.1 microM Trichostatin A to 2.3 at 1 microM Trichostatin A. Similarly, caspase-3 and caspase-7 assays indicated that Trichostatin A potentiates radiation-induced apoptosis in K562 cells, in a concentration dependent manner. Our results suggest the modulation of radiation effects observed at the lower Trichostatin A concentrations was associated with histone hyperacetylation and changes in phosphorylated gammaH2A.X formation on euchromatin. In contrast, at the higher Trichostatin A concentrations mechanisms such as drug-mediated cytotoxicity and G1 cell cycle arrest, contributed to the sensitization effect. More generally, our findings are consistent with those from recent studies and support the development of histone deacetylase inhibitors for use as radiation sensitizers, particularly for targeting radioresistant cancers.     

Predicting COVID-19 Transmission to Inform the Management of Mass Events: Model-Based Approach

BackgroundModelling COVID-19 transmission at live events and public gatherings is essential to controlling the probability of subsequent outbreaks and communicating to participants their personalized risk. Yet, despite the fast-growing body of literature on COVID-19 transmission dynamics, current risk models either neglect contextual information including vaccination rates or disease prevalence or do not attempt to quantitatively model transmission.ObjectiveThis paper attempted to bridge this gap by providing informative risk metrics for live public events, along with a measure of their uncertainty.MethodsBuilding upon existing models, our approach ties together 3 main components: (1) reliable modelling of the number of infectious cases at the time of the event, (2) evaluation of the efficiency of pre-event screening, and (3) modelling of the event’s transmission dynamics and their uncertainty using Monte Carlo simulations.ResultsWe illustrated the application of our pipeline for a concert at the Royal Albert Hall and highlighted the risk’s dependency on factors such as prevalence, mask wearing, and event duration. We demonstrate how this event held on 3 different dates (August 20, 2020; January 20, 2021; and March 20, 2021) would likely lead to transmission events that are similar to community transmission rates (0.06 vs 0.07, 2.38 vs 2.39, and 0.67 vs 0.60, respectively). However, differences between event and background transmissions substantially widened in the upper tails of the distribution of the number of infections (as denoted by their respective 99th quantiles: 1 vs 1, 19 vs 8, and 6 vs 3, respectively, for our 3 dates), further demonstrating that sole reliance on vaccination and antigen testing to gain entry would likely significantly underestimate the tail risk of the event.ConclusionsDespite the unknowns surrounding COVID-19 transmission, our estimation pipeline opens the discussion on contextualized risk assessment by combining the best tools at hand to assess the order of magnitude of the risk. Our model can be applied to any future event and is presented in a user-friendly RShiny interface. Finally, we discussed our model’s limitations as well as avenues for model evaluation and improvement.     

Deep sequencing reveals increased DNA methylation in chronic rat epilepsy

Epilepsy is a frequent neurological disorder, although onset and progression of seizures remain difficult to predict in affected patients, irrespective of their epileptogenic condition. Previous studies in animal models as well as human epileptic brain tissue revealed a remarkably diverse pattern of gene expression implicating epigenetic changes to contribute to disease progression. Here we mapped for the first time global DNA methylation patterns in chronic epileptic rats and controls. Using methyl-CpG capture associated with massive parallel sequencing (Methyl-Seq) we report the genomic methylation signature of the chronic epileptic state. We observed a predominant increase, rather than loss of DNA methylation in chronic rat epilepsy. Aberrant methylation patterns were inversely correlated with gene expression changes using mRNA sequencing from same animals and tissue specimens. Administration of a ketogenic, high-fat, low-carbohydrate diet attenuated seizure progression and ameliorated DNA methylation mediated changes in gene expression. This is the first report of unsupervised clustering of an epigenetic mark being used in epilepsy research to separate epileptic from non-epileptic animals as well as from animals receiving anti-convulsive dietary treatment. We further discuss the potential impact of epigenetic changes as a pathogenic mechanism of epileptogenesis.     

In vitro model for the study of human posterior capsule opacification

PURPOSE: To develop and evaluate a model for the organ culture of human lens capsules that reduces problems inherent in preexisting models for the study of in vitro posterior capsule opacification (PCO). METHODS: Human lenses (N = 110) were isolated from donor eyes and supported externally within a lens holder system by medical-grade cyanoacrylate glue, allowing visualization of the entire capsular bag. After capsulorhexis and lens extraction were performed, the capsule specimens were maintained at physiological conditions for up to 4 weeks. The area of lens epithelial cell (LEC) coverage over the posterior capsule surface was determined objectively on a daily basis using a graticule. Lens epithelial cell behavior was correlated with clinical data and other in vitro PCO models. RESULTS: Cyanoacrylate glue did not appear to be toxic to LECs at the concentration used. The amount of viable epithelium after nuclear extraction was dependent on the age and postmortem time of the specimen. Viable LEC cultures were obtained from eyes up to 9 days postmortem. The time from death to culture or from enucleation to culture did not influence LEC viability if it was fewer than 5 days. The LEC proliferation rates and confluence times were age dependent and correlated closely between pairs of eyes. CONCLUSIONS: Results show that the lens holder model is a more physiological method for supporting the capsule and is a robust, reproducible system for the study of LEC migration and proliferation. It allows visualization within the entire capsular bag. Intraocular lenses can be implanted in this system in a way that more closely resembles the in vivo scenario. This model can be used to evaluate therapeutic measures to prevent PCO.     

Epigenetic phenomena linked to diabetic complications

Diabetes mellitus (type 1 and type 2) and the complications associated with this condition are an urgent public health problem, as the incidence of diabetes mellitus is steadily increasing. Environmental factors, such as diet and exposure to hyperglycemia, contribute to the etiology of diabetes mellitus and its associated microvascular and macrovascular complications. These vascular complications are the main cause of the morbidity and mortality burden of diabetes mellitus. The DCCT-EDIC and UKPDS epidemiological studies correlated poor glycemic control with the development of vascular complications in patients with type 1 or type 2 diabetes mellitus. The findings of these studies suggest that early exposure to hyperglycemia predisposes individuals to the development of diabetic complications, a phenomenon referred to as metabolic memory or the legacy effect. The first experimental evidence for metabolic memory was reported >20 years ago and the underlying molecular mechanisms are currently being characterized. Interestingly, transient exposure to hyperglycemia results in long-lasting epigenetic modifications that lead to changes in chromatin structure and gene expression, which mediate these persistent metabolic characteristics.     

Failure of a discontinuous bend to prevent lens epithelial cell migration in vitro

PURPOSE: To assess the effect of substrate geometry (discontinuous bend) on lens epithelial cell (LEC) growth in vitro. SETTING: Department of Ophthalmology, St. Thomas' Hospital, London, United Kingdom. METHODS: Culture wells with central depths of 0.4 mm, 1.0 mm, or 3.0 mm and a sharp square-edged profile (discontinuous bend) or a round-edged profile (continuous bend) were produced from a block of poly(methyl methacrylate). Freshly harvested bovine LECs were attached to the center of each well and cultured using standard techniques. Observations were made of whether LECs grew out of the wells and of the time required to do so. RESULTS: Lens epithelial cells migrated out of all the wells. There was no significant difference in the rate at which they migrated out of round-edged and square-edged wells. CONCLUSIONS: In vitro, a sharp discontinuous bend did not appear to induce contact inhibition of migrating LECs nor did it significantly hinder the rate at which LECs migrated. Therefore, a discontinuous bend in the lens capsule in isolation is unlikely to be responsible for the observed reduction in posterior capsule opacification associated with the use of square-edged IOLs.     

Mathematical modeling of the forces between an intraocular lens and the capsule

PURPOSE: To model the pressure relationship between an intraocular lens (IOL) and the capsular bag. SETTING: Department of Physics, Kings' College, London University, London, United Kingdom. METHODS: A mathematical model was made of the forces between the capsule and IOL showing that the pressure related to the local radius of curvature of the IOL at any given point. The local radius of curvature for round-edged and square-edged IOLs was measured from electron micrographs of the IOL profiles, and the corresponding pressure profiles were calculated and compared. RESULTS: The pressure between an IOL and the capsular bag was proportional to the quotient of the tension in the capsule divided by the local radius of curvature of the IOL, with a constant of proportionality that depended on the coefficient of friction between the capsule and IOL. Measuring the local radius of curvature of the 2 IOL types suggested a pressure increase of at least 69% +/- 6% at the optic edge with the square-edged IOL. CONCLUSIONS: The mathematical model predicted that IOLs with square-edged optic profiles exerts higher pressure on the posterior capsule than IOLs with round-edged optic profiles. The higher pressure may form a physical barrier to lens epithelial cell migration onto the posterior capsule.