Freeze-Drying of Mononuclear Cells Derived from Umbilical Cord Blood Followed by Colony Formation

BACKGROUND: We recently showed that freeze-dried cells stored for 3 years at room temperature can direct embryonic development following cloning. However, viability, as evaluated by membrane integrity of the cells after freeze-drying, was very low; and it was mainly the DNA integrity that was preserved. In the present study, we improved the cells' viability and functionality after freeze-drying. METHODOLOGY/PRINCIPAL FINDINGS: We optimized the conditions of directional freezing, i.e. interface velocity and cell concentration, and we added the antioxidant EGCG to the freezing solution. The study was performed on mononuclear cells (MNCs) derived from human umbilical cord blood. After freeze-drying, we tested the viability, number of CD34(+)-presenting cells and ability of the rehydrated hematopoietic stem cells to differentiate into different blood cells in culture. The viability of the MNCs after freeze-drying and rehydration with pure water was 88%-91%. The total number of CD34(+)-presenting cells and the number of colonies did not change significantly when evaluated before freezing, after freeze-thawing, and after freeze-drying (5.4 x 10(4)+/-4.7, 3.49 x 10(4)+/-6 and 6.31 x 10(4)+/-12.27 cells, respectively, and 31+/-25.15, 47+/-45.8 and 23.44+/-13.3 colonies, respectively). CONCLUSIONS: This is the first report of nucleated cells which have been dried and then rehydrated with double-distilled water remaining viable, and of hematopoietic stem cells retaining their ability to differentiate into different blood cells

Topoisomer Differentiation of Molecular Knots by FTICR MS: Lessons from Class II Lasso Peptides

Lasso peptides constitute a class of bioactive peptides sharing a knotted structure where the C-terminal tail of the peptide is threaded through and trapped within an N-terminalmacrolactamring. The structural characterization of lasso structures and differentiation from their unthreaded topoisomers is not trivial and generally requires the use of complementary biochemical and spectroscopic methods. Here we investigated two antimicrobial peptides belonging to the class II lasso peptide family and their corresponding unthreaded topoisomers: microcin J25 (MccJ25), which is known to yield two-peptide product ions specific of the lasso structure under collisioninduced dissociation (CID), and capistruin, for which CID does not permit to unambiguously assign the lasso structure. The two pairs of topoisomers were analyzed by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS) upon CID, infrared multiple photon dissociation (IRMPD), and electron capture dissociation (ECD). CID and ECDspectra clearly permitted to differentiate MccJ25 from its non-lasso topoisomer MccJ25-Icm, while for capistruin, only ECD was informative and showed different extent of hydrogen migration (formation of c\bullet/z from c/z\bullet) for the threaded and unthreaded topoisomers. The ECD spectra of the triply-charged MccJ25 and MccJ25-lcm showed a series of radical b-type product ions {\eth}b0In{\TH}. We proposed that these ions are specific of cyclic-branched peptides and result from a dual c/z\bullet and y/b dissociation, in the ring and in the tail, respectively. This work shows the potentiality of ECD for structural characterization of peptide topoisomers, as well as the effect of conformation on hydrogen migration subsequent to electron capture

Computable analysis of linear rearrangement optimization

Optimization problems over rearrangement classes arise in various areas such as mathematics, fluid mechanics, biology, and finance. When the generator of the rearrangement class is two-valued, they reduce to shape optimization and free boundary problems which can exhibit intriguing symmetry breaking phenomena. A robust framework is required for computable analysis of these problems. In this paper, as a first step towards such a robust framework, we provide oracle Turing machines that compute the distribution function, decreasing rearrangement, and linear rearrangement optimizers, with respect to functions that are continuous and have no significant flat zones. This assumption on the reference function is necessary, as otherwise, the aforementioned operations may not be computable. We prove that the results can be computed to within any degree of accuracy, conforming to the framework of Type-II Theory of Effectivity

Practicing convict criminology: lessons learned from British academic activism

Joanne Belknap’s recent ASC presidential address included a critique of Convict Criminology’s activism. A number of concerns were provided, although of particular importance here are, first, Belknap’s concerns regarding the absence of ‘marginalized voices’ in the Convict Criminology network. Second, the issue of defining how non-con academics function as Convict Criminology group members. This paper responds to these criticisms. Specifically, we discuss the question of ‘representation’ in BCC and our attempts to remedy this issue. We also draw attention to the academic activism that British Convict Criminology is conducting in Europe. This includes a detailed discussion of the collaborative research-activist activities that involve non-con as well as ex-con academic network members. We demonstrate how these collaborations explain the vital group membership role that non-con academics assume in the activism of Convict Criminology

The road to hell is paved with good intentions: the experience of applying for national data for linkage and suggestions for improvement

Background We can improve healthcare services by better understanding current provision. One way to understand this is by linking data sets from clinical and national audits, national registries and other National Health Service (NHS) encounter data. However, getting to the point of having linked national data sets is challenging. Objective We describe our experience of the data application and linkage process for our study ‘LAUNCHES QI’, and the time, processes and resource requirements involved. To help others planning similar projects, we highlight challenges encountered and advice for applications in the current system as well as suggestions for system improvements. Findings The study set up for LAUNCHES QI began in March 2018, and the process through to data acquisition took 2.5 years. Several challenges were encountered, including the amount of information required (often duplicate information in different formats across applications), lack of clarity on processes, resource constraints that limit an audit’s capacity to fulfil requests and the unexpected amount of time required from the study team. It is incredibly difficult to estimate the resources needed ahead of time, and yet necessary to do so as early on as funding applications. Early decisions can have a significant impact during latter stages and be hard to change, yet it is difficult to get specific information at the beginning of the process. Conclusions The current system is incredibly complex, arduous and slow, stifling innovation and delaying scientific progress. NHS data can inform and improve health services and we believe there is an ethical responsibility to use it to do so. Streamlining the number of applications required for accessing data for health services research and providing clarity to data controllers could facilitate the maintenance of stringent governance, while accelerating scientific studies and progress, leading to swifter application of findings and improvements in healthcare

A systematic review and meta-synthesis of the impact of low back pain on people's lives

Copyright @ 2014 Froud et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.Background - Low back pain (LBP) is a common and costly problem that many interpret within a biopsychosocial model. There is renewed concern that core-sets of outcome measures do not capture what is important. To inform debate about the coverage of back pain outcome measure core-sets, and to suggest areas worthy of exploration within healthcare consultations, we have synthesised the qualitative literature on the impact of low back pain on people’s lives. Methods - Two reviewers searched CINAHL, Embase, PsycINFO, PEDro, and Medline, identifying qualitative studies of people’s experiences of non-specific LBP. Abstracted data were thematic coded and synthesised using a meta-ethnographic, and a meta-narrative approach. Results - We included 49 papers describing 42 studies. Patients are concerned with engagement in meaningful activities; but they also want to be believed and have their experiences and identity, as someone ‘doing battle’ with pain, validated. Patients seek diagnosis, treatment, and cure, but also reassurance of the absence of pathology. Some struggle to meet social expectations and obligations. When these are achieved, the credibility of their pain/disability claims can be jeopardised. Others withdraw, fearful of disapproval, or unable or unwilling to accommodate social demands. Patients generally seek to regain their pre-pain levels of health, and physical and emotional stability. After time, this can be perceived to become unrealistic and some adjust their expectations accordingly. Conclusions - The social component of the biopsychosocial model is not well represented in current core-sets of outcome measures. Clinicians should appreciate that the broader impact of low back pain includes social factors; this may be crucial to improving patients’ experiences of health care. Researchers should consider social factors to help develop a portfolio of more relevant outcome measures.Arthritis Research U

Turbulence and aeration in hydraulic jumps: free-surface fluctuation and integral turbulent scale measurements

In an open channel, a change from a supercritical to subcritical flow is a strong dissipative process called a hydraulic jump. Herein some new measurements of free-surface fluctuations of the impingement perimeter and integral turbulent time and length scales in the roller are presented with a focus on turbulence in hydraulic jumps with a marked roller. The observations highlighted the fluctuating nature of the impingement perimeter in terms of both longitudinal and transverse locations. The results showed further the close link between the production and detachment of large eddies in jump shear layer, and the longitudinal fluctuations of the jump toe. They highlighted the importance of the impingement perimeter as the origin of the developing shear layer and a source of vorticity. The air–water flow measurements emphasised the intense flow aeration. The turbulent velocity distributions presented a shape similar to a wall jet solution with a marked shear layer downstream of the impingement point. The integral turbulent length scale distributions exhibited a monotonic increase with increasing vertical elevation within 0.2 < Lz/d1 < 0.8 in the shear layer, where Lz is the integral turbulent length scale and d1 the inflow depth, while the integral turbulent time scales were about two orders of magnitude smaller than the period of impingement position longitudinal oscillations