Translational and Regulatory Challenges for Exon Skipping Therapies

Several translational challenges are currently impeding the therapeutic development of antisense-mediated exon skipping approaches for rare diseases. Some of these are inherent to developing therapies for rare diseases, such as small patient numbers and limited information on natural history and interpretation of appropriate clinical outcome measures. Others are inherent to the antisense oligonucleotide (AON)-mediated exon skipping approach, which employs small modified DNA or RNA molecules to manipulate the splicing process. This is a new approach and only limited information is available on long-term safety and toxicity for most AON chemistries. Furthermore, AONs often act in a mutation-specific manner, in which case multiple AONs have to be developed for a single disease. A workshop focusing on preclinical development, trial design, outcome measures, and different forms of marketing authorization was organized by the regulatory models and biochemical outcome measures working groups of Cooperation of Science and Technology Action: "Networking towards clinical application of antisense-mediated exon skipping for rare diseases." The workshop included participants from patient organizations, academia, and members of staff from the European Medicine Agency and Medicine Evaluation Board (the Netherlands). This statement article contains the key outcomes of this meeting.status: publishe

On automated sequential steady-state simulation.

The credibility of the final results from stochastic simulation has had limited discussion in the simulation literature so far. However, it is important that the final results from any simulations be credible. To achieve this, validation, which determines whether the conceptual simulation model is an accurate representation of the system under study, has to be done carefully. Additionally, a proper statistical analysis of simulation output data, including a confidence interval or other assessment of statistical errors, has to be conducted before any valid inferences or conclusions about the performance of simulated dynamic systems, such as for example telecommunication networks, are made. There are many other issues, such as choice of a good pseudo-random number generator, elimination of initialisation bias in steady-state simulations, and consideration of auto correlations in collected observations, which have to be appropriately addressed for the final results to be credible. However, many of these issues are not trivial, particularly for simulation users who may not be experts in these areas. As a consequence, a fully-automated simulation package, which can control all important aspects of stochastic simulation, is needed. This dissertation focuses on the following contributions to such a package for steady-state simulation: properties of confidence intervals (CIs) used in coverage analysis, heuristic rules for improving the coverage of the final CIs in practical applications, automated sequential analysis of mean values by the method of regenerative cycles, automatic detection of the initial transient period for steady-state quantile estimation, and sequential steady-state quantile estimation with the automated detection of the length of initial transient period. One difficulty in obtaining precise estimates of a system using stochastic simulation can be the cost of the computing time needed to collect the large amount of output data required. Indeed there are situations, such as estimation of rare events, where, even assuming an appropriate statistical analysis procedure is available, the cost of collecting the number of observations needed by the analysis procedure can be prohibitively large. Fortunately, inexpensive computer network resources enable computationally intensive simulations by allowing us to run parallel and distributed simulations. Therefore, where possible, we extend the contributions to the distributed stochastic simulation scenario known as the Multiple Replications In Parallel (MRIP), in which multiple processors run their own independent replications of the simulated system but cooperate with central analysers that collect data to estimate the final results

Optimization of a Do-It-Yourself Air Cleaner Design to Reduce Residential Air Pollution Exposure for a Community Experiencing Environmental Injustices

&nbsp; The large-scale deployment of Do-it-yourself (DIY) air cleaners, especially in communities that historically bear the brunt of air pollution exposure-related injustices, provides communities a cost-effective option to reduce personal indoor exposure to particulate matter. In this study, we developed nine air cleaner prototypes, altering filter depth and the number and type of filters, and compared their PM2.5 removal effectiveness and maintenance-related parameters prior to deployment in North Denver, Colorado homes. Prototypes containing multiple high efficiency particulate air filters with a minimum reporting value of 13 (MERV13) had higher clean air delivery rates (CADR, &gt;300 m3 h&minus;1) compared to prototypes using a single filter (100&ndash;200 m3 h&minus;1), but single-filter designs had comparable values of CADR normalized by initial and annual operating costs. Based on performance, cost, build time, and feedback from the community regarding concerns related to volatile organic compound exposure, the selected prototype (P9) used a combination of an activated carbon filter and single MERV13 filter with a 10.16 cm (4-inch) depth. Following this assessment, 120 of the selected air cleaner prototypes were built and deployed in homes around the communities in North Denver for two separate cohorts; feedback regarding their usage over the course of the deployment showed that in addition to the increased noise levels perceived by the participants, factors such as cold air flow from the air cleaner impacting the thermal comfort and aesthetics of the design reduced their usage time in homes. Future designs of DIY air cleaners could incorporate this feedback to help design improved features such as quieter air cleaners and real-time pollutant monitoring feedback to prompt users to keep them operational at all times of the day.</div

Prevalence of diabetic retinopathy in Tehran province: a population-based study

<p>Abstract</p> <p>Background</p> <p>To determine the prevalence and characteristics of diabetic retinopathy (DR) among Iranian patients with diabetes.</p> <p>Methods</p> <p>Design: population-based cross-sectional study.</p> <p>Participants: patients with diabetes aged 25 to 64 years in Tehran province, Iran. This survey was conducted from April to October 2007. The study sample was derived from the first national survey of risk factors for non-communicable disease. Diabetes mellitus was defined as a fasting plasma glucose of ≥ 7.0 mmol/l (126 mg/dl) or more, use of diabetic medications, or a physician's diagnosis of diabetes. All patients known to have diabetes underwent an eye examination by bio-microscope and indirect ophthalmoscope to check for any signs of DR through dilated pupils by + 78 lens. Participants were also interviewed and examined to determine their demographic characteristics, medical conditions and the regularity of their eye visits.</p> <p>Results</p> <p>Among 7989 screened patients, 759 (9.5%) had diabetes. Of them, 639 patients (84.2%) underwent eye examination. Five patients (0.7%) with media opacity were excluded. Of 634 examined patients with diabetes, 240 had some degree of diabetic retinopathy, and the overall standardized prevalence of any retinopathy was 37.0% (95% CI: 33.2-40.8), including 27.3% (95% CI: 23.7-30.8) (n = 175) with non-proliferative and 9.6% (95% CI: 7.3-11.9) (n = 65) with proliferative diabetic retinopathy. Clinically significant macular edema and vision-threatening retinopathy were detected in 5.8% (95% CI: 4.0-7.7) (n = 38) and 14.0% (95% CI: 11.3-16.7) (n = 95) of patients, respectively. Only 143 patients (22.6%) with diabetes had a history of regular eye examination.</p> <p>Conclusion</p> <p>This study demonstrated a high prevalence and poor control of DR in Tehran province. This suggests the need for adequate prevention and treatment in patients with diabetes.</p

Ectopic T Cell Receptor-α Locus Control Region Activity in B Cells Is Suppressed by Direct Linkage to Two Flanking Genes at Once

The molecular mechanisms regulating the activity of the TCRα gene are required for the production of the circulating T cell repertoire. Elements of the mouse TCRα locus control region (LCR) play a role in these processes. We previously reported that TCRα LCR DNA supports a gene expression pattern that mimics proper thymus-stage, TCRα gene-like developmental regulation. It also produces transcription of linked reporter genes in peripheral T cells. However, TCRα LCR-driven transgenes display ectopic transcription in B cells in multiple reporter gene systems. The reasons for this important deviation from the normal TCRα gene regulation pattern are unclear. In its natural locus, two genes flank the TCRα LCR, TCRα (upstream) and Dad1 (downstream). We investigated the significance of this gene arrangement to TCRα LCR activity by examining transgenic mice bearing a construct where the LCR was flanked by two separate reporter genes. Surprisingly, the presence of a second, distinct, reporter gene downstream of the LCR virtually eliminated the ectopic B cell expression of the upstream reporter observed in earlier studies. Downstream reporter gene activity was unaffected by the presence of a second gene upstream of the LCR. Our findings indicate that a gene arrangement in which the TCRα LCR is flanked by two distinct transcription units helps to restrict its activity, selectively, on its 5′-flanking gene, the natural TCRα gene position with respect to the LCR. Consistent with these findings, a TCRα/Dad1 locus bacterial artificial chromosome dual-reporter construct did not display the ectopic upstream (TCRα) reporter expression in B cells previously reported for single TCRα transgenes

VizieR Online Data Catalog: 3C388 145, 392, 614, 1400 and 4850MHz images (Brienza+, 2020)

We used a recent dataset obtained on June 26th, 2019, as part of the LOFAR Two-metre Sky Survey (LoTSS, see Shimwell et al., 2019A&A...622A...1S, Cat. J/A+A/622/A1). We observed the source with the Very Large Array (VLA) in A configuration on July 28th 2015 using the P-band receiver centered at 350MHz. We reprocessed the data used by Roettiger et al. (1994ApJ...421L..23R) at 1400MHz and 4850MHz. The data consists of observations in B array at 1400MHz and in C array at 4850MHz. The target was observed for 7 hours at 1400MHz and for 5 hours at 4850MHz. The target was observed with the legacy Giant Metrewave Radio Telescope (GMRT) at 614MHz and 240MHz in dual frequency mode and data were published in Lal et al. (2008MNRAS.390.1105L). The observations were performed on July 29th and 30th, 2005. 3C388 was observed by Chandra on February 9th and 29th, 2004 with the ACIS-I detector (obs ID 4756 and 5295, respectively) and the data were published by Kraft et al. (2006ApJ...639..753K). (2 data files)

Transcriptional regulation of Elf-1: locus-wide analysis reveals four distinct promoters, a tissue-specific enhancer, control by PU.1 and the importance of Elf-1 downregulation for erythroid maturation

Ets transcription factors play important roles during the development and maintenance of the haematopoietic system. One such factor, Elf-1 (E74-like factor 1) controls the expression of multiple essential haematopoietic regulators including Scl/Tal1, Lmo2 and PU.1. However, to integrate Elf-1 into the wider regulatory hierarchies controlling haematopoietic development and differentiation, regulatory elements as well as upstream regulators of Elf-1 need to be identified. Here, we have used locus-wide comparative genomic analysis coupled with chromatin immunoprecipitation (ChIP-chip) assays which resulted in the identification of five distinct regulatory regions directing expression of Elf-1. Further, ChIP-chip assays followed by functional validation demonstrated that the key haematopoietic transcription factor PU.1 is a major upstream regulator of Elf-1. Finally, overexpression studies in a well-characterized erythroid differentiation assay from primary murine fetal liver cells demonstrated that Elf-1 downregulation is necessary for terminal erythroid differentiation. Given the known activation of PU.1 by Elf-1 and our newly identified reciprocal activation of Elf-1 by PU.1, identification of an inhibitory role for Elf-1 has significant implications for our understanding of how PU.1 controls myeloid–erythroid differentiation. Our findings therefore not only represent the first report of Elf-1 regulation but also enhance our understanding of the wider regulatory networks that control haematopoiesis

Imminent brain death: point of departure for potential heart-beating organ donor recognition

Contains fulltext : 88186.pdf (publisher's version ) (Closed access)PURPOSE: There is, in European countries that conduct medical chart review of intensive care unit (ICU) deaths, no consensus on uniform criteria for defining a potential organ donor. Although the term is increasingly being used in recent literature, it is seldom defined in detail. We searched for criteria for determination of imminent brain death, which can be seen as a precursor for organ donation. METHODS: We organized meetings with representatives from the field of clinical neurology, neurotraumatology, intensive care medicine, transplantation medicine, clinical intensive care ethics, and organ procurement management. During these meetings, all possible criteria were discussed to identify a patient with a reasonable probability to become brain dead (imminent brain death). We focused on the practical usefulness of two validated coma scales (Glasgow Coma Scale and the FOUR Score), brain stem reflexes and respiration to define imminent brain death. Further we discussed criteria to determine irreversibility and futility in acute neurological conditions. RESULTS: A patient who fulfills the definition of imminent brain death is a mechanically ventilated deeply comatose patient, admitted to an ICU, with irreversible catastrophic brain damage of known origin. A condition of imminent brain death requires either a Glasgow Coma Score of 3 and the progressive absence of at least three out of six brain stem reflexes or a FOUR score of E(0)M(0)B(0)R(0). CONCLUSION: The definition of imminent brain death can be used as a point of departure for potential heart-beating organ donor recognition on the intensive care unit or retrospective medical chart analysis.1 september 201