Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions

Regardless of their tissue of origin, multipotent mesenchymal stromal cells (MSCs) are commonly expanded in vitro for several population doublings, in order to achieve a sufficient number of cells for therapy. Prolonged MSC expansion has shown to result in phenotypical, morphological and gene expression changes in MSCs, which ultimately lead to the state of senescence. The presence of senescent cells in therapeutic MSC batches is undesirable, as it reduces their viability, differentiation potential and trophic capabilities. Additionally, senescent cells acquire senescence-activated secretory phenotype, which may not only induce apoptosis in the neighbouring host cells following MSC transplantation, but also trigger local inflammatory reactions. This review outlines the current and promising new methodologies for the identification of senescent cells in MSC cultures, with a particular emphasis on non-destructive and label-free methodologies. Technologies allowing identification of individual senescent cells, based on new surface markers, offer potential advantage for targeted senescent cell removal using new-generation senolytic agents, and subsequent production of therapeutic MSC batches fully devoid of senescent cells. Methods or a combination of methods that are non-destructive and label-free, for example involving cell size and spectroscopic measurements, could be the best way forward as they do not modify the cells of interest thus maximising the final output of therapeutic-grade MSC cultures. The further incorporation of machine learning methods has also recently shown promise in facilitating, automating and enhancing the analysis of these measured data

Quasilocal energy for rotating charged black hole solutions in general relativity and string theory

We explore the (non)-universality of Martinez's conjecture, originally proposed for Kerr black holes, within and beyond general relativity. The conjecture states that the Brown-York quasilocal energy at the outer horizon of such a black hole reduces to twice its irreducible mass, or equivalently, to \sqrt{A} /(2\sqrt{pi}), where `A' is its area. We first consider the charged Kerr black hole. For such a spacetime, we calculate the quasilocal energy within a two-surface of constant Boyer-Lindquist radius embedded in a constant stationary-time slice. Keeping with Martinez's conjecture, at the outer horizon this energy equals the irreducible mass. The energy is positive and monotonically decreases to the ADM mass as the boundary-surface radius diverges. Next we perform an analogous calculation for the quasilocal energy for the Kerr-Sen spacetime, which corresponds to four-dimensional rotating charged black hole solutions in heterotic string theory. The behavior of this energy as a function of the boundary-surface radius is similar to the charged Kerr case. However, we show that in this case it does not approach the expression conjectured by Martinez at the horizon.Comment: 15 page

Single-vehicle collisions in Europe: analysis using real-world and crash-test data

A large proportion of European road casualties result when a vehicle leaves the main carriageway, often impacting roadside obstacles. As part of the EC-funded project, RISER (Roadside Infrastructure for Safer European Roads), a number of activities were undertaken to collate the type of data which is needed to understand the frequency and severity of real world crash situations and relate this to crash test data mandated in the EU. Accident data was collected and used to create a statistical database and a detailed database exclusively for single vehicle 'run-off the road' collisions on major rural (not urban) roads, simulation software was used to further understand impacts with roadside structures and an inventory of crash test data was collected for impacts with objects such as poles and safety barriers . The combination of real world accident data, simulations and crash test data has provided a unique insight into the characteristics of single vehicle collisions, helping us to understand them better and make recommendations for consideration when drafting design guidelines. This information is crucial for those involved in the design and evaluation of the roadside environment

Genomic Investigation of Two Acinetobacter baumannii Outbreaks in a Veterinary Intensive Care Unit in The Netherlands

Acinetobacter baumannii is a nosocomial pathogen that frequently causes healthcare-acquired infections. The global spread of multidrug-resistant (MDR) strains with its ability to survive in the environment for extended periods imposes a pressing public health threat. Two MDR A. baumannii outbreaks occurred in 2012 and 2014 in a companion animal intensive care unit (caICU) in the Netherlands. Whole-genome sequencing (WGS) was performed on dog clinical isolates (n = 6), environmental isolates (n = 5), and human reference strains (n = 3) to investigate if the isolates of the two outbreaks were related. All clinical isolates shared identical resistance phenotypes displaying multidrug resistance. Multi-locus Sequence Typing (MLST) revealed that all clinical isolates belonged to sequence type ST2. The core genome MLST (cgMLST) results confirmed that the isolates of the two outbreaks were not related. Comparative genome analysis showed that the outbreak isolates contained different gene contents, including mobile genetic elements associated with antimicrobial resistance genes (ARGs). The time-measured phylogenetic reconstruction revealed that the outbreak isolates diverged approximately 30 years before 2014. Our study shows the importance of WGS analyses combined with molecular clock investigations to reduce transmission of MDR A. baumannii infections in companion animal clinics

Trip related factors

WP3 of the European Project TRACE is concerned with Types of Factors to analyse the causation of road traffic accidents from a factors' point of view. In task 3.3 'Trip-related Factors' it was tried to characterise accidents that are caused by certain contributing factors found on a trip level. This was done by applying one statistical method to existing databases of the WP3 Partners on the one hand and on the other hand by performing an in-depth case analysis using the WP5 method. The analysed factors stem from the Human Component of the accident causation classification, namely "alcohol", "vigilance", and "experience", from the Vehicle Component, namely "vehicle condition/maintenance", and from the Environment component, namely "road layout" and "road condition". This selection resulted from the task 3.1 conclusions and feasibility reasons. Due to inhomogeneous results for the databases from Austria, France, Germany, Great Britain, and Spain the detailed results will be pictured in an Internal TRACE Report by Sub-reports of the WP3 Partners, in this task report the main results are discussed with respect to findings and data in other databases available to the TRACE partners as requested from WP8. Both methods applied show that trip-related factors are possible to prevent not on a trip level only, but also from a background level and as well on a level closer to the accident (driving task level). However, only some suggestions are possible to give by these results. A more detailed view for preventing the different accidents that result from trip-related factors is necessary, as shown by the results of BASt with the statistic method, as well as by INRETS with the ultra in-depth WP5 method

Summary report on work package 3 "Types of Factors"

This summary report presents the main results of Work Package 3 "Types of Factors" of the TRACE Project. The work as performed in the tasks 3.1 (accident related factors), 3.2 (sociological and cultural factors), 3.3 (trip-related factors), and 3.4 (driving-task associated factors) and presented in the Deliverables 3.1 to 3.4 and an additional internal TRACE Report (Collection of Sub-Reports for task 3.3) is summarized and discussed. The objective of defining relevant accident related factors first and the objective of analysing traffic accident causation - from a factor's point of view while taking traditional views into account - on different levels - by using statistic methods for existing databases as provided by the Work Package 3 Partners and - by using new (developed in Work Package 5 of the TRACE project) methods on new case analysis in order to gain new knowledge on accident causation was possible to reach. The scope of the identified key aspects as found by the Partners in their work for the relevance in EU27 is discussed. In accordance, even further, appropriate suggestions for prevention of traffic accidents can be derived