Establishing Clonal Cell Lines with Endothelial-Like Potential from CD9(hi), SSEA-1(-) Cells in Embryonic Stem Cell-Derived Embryoid Bodies

Background. Differentiation of embryonic stem cells (ESCs) into specific cell types with minimal risk of teratoma formation could be efficiently directed by first reducing the differentiation potential of ESCs through the generation of clonal, self-renewing lineage-restricted stem cell lines. Efforts to isolate these stem cells are, however, mired in an impasse where the lack of purified lineage-restricted stem cells has hindered the identification of defining markers for these rare stem cells and, in turn, their isolation. Methodology/Principal Findings. We describe here a method for the isolation of clonal lineage-restricted cell lines with endothelial potential from ESCs through a combination of empirical and rational evidence-based methods. Using an empirical protocol that we have previously developed to generate embryo-derived RoSH lines with endothelial potential, we first generated E-RoSH lines from mouse ESC-derived embryoid bodies (EBs). Despite originating from different mouse strains, RoSH and E-RoSH lines have similar gene expression profiles (r(2) = 0.93) while that between E-RoSH and ESCs was 0.83. In silico gene expression analysis predicted that like RoSH cells, E-RoSH cells have an increased propensity to differentiate into vasculature. Unlike their parental ESCs, E-RoSH cells did not form teratomas and differentiate efficiently into endothelial-like cells in vivo and in vitro. Gene expression and FACS analysis revealed that RoSH and E-RoSH cells are CD9(hi), SSEA-1(-) while ESCs are CD9(lo), SSEA-1(+). Isolation of CD9(hi), SSEA-1(-) cells that constituted 1%-10% of EB-derived cultures generated an E-RoSH-like culture with an identical E-RoSH-like gene expression profile (r(2) = 0.95) and a propensity to differentiate into endothelial-like cells. Conclusions. By combining empirical and rational evidence-based methods, we identified definitive selectable surface antigens for the isolation and propagation of lineage-restricted stem cells with endothelial-like potential from mouse ESCs.

Towards accurate estimation of the proportion of true null hypotheses in multiple testing

Background

Biomedical researchers are now often faced with situations where it is necessary to test a large number of hypotheses simultaneously, eg, in comparative gene expression studies using high-throughput microarray technology. To properly control false positive errors the FDR (false discovery rate) approach has become widely used in multiple testing. The accurate estimation of FDR requires the proportion of true null hypotheses being accurately estimated. To date many methods for estimating this quantity have been proposed. Typically when a new method is introduced, some simulations are carried out to show the improved accuracy of the new method. However, the simulations are often very limited to covering only a few points in the parameter space.

Results

Here I have carried out extensive in silico experiments to compare some commonly used methods for estimating the proportion of true null hypotheses. The coverage of these simulations is unprecedented thorough over the parameter space compared to typical simulation studies in the literature. Thus this work enables us to draw conclusions globally as to the performance of these different methods. It was found that a very simple method gives the most accurate estimation in a dominantly large area of the parameter space. Given its simplicity and its overall superior accuracy I recommend its use as the first choice for estimating the proportion of true null hypotheses in multiple testing.

The Role of Self-Help Efforts in the Reintegration of ‘Politically Motivated’ Former Prisoners: Implications from the Northern Irish experience

Self-help (or mutual aid) processes play a substantial role in the reintegration of stigmatized individuals, in particular, a substantial self-help movement has developed around addiction recovery. Prisoners and ex-prisoners have also established self-help groups around the world. This paper focuses in particular on the role of self-help principles and practices among “politically motivated” former prisoners from all sides of the Northern Irish conflict. The concept of self-help and its application to former prisoners are analysed theoretically, then applied to the Northern Irish case study through a series of interviews with ex-prisoners whose incarceration has been related to the conflict in Northern Ireland. We draw on the implications of this case study for wider issues of reintegration for politically motivated and ordinary prisoners.

Genetic Evidence Implicates the Immune System and Cholesterol Metabolism in the Aetiology of Alzheimer's Disease

Background: Late Onset Alzheimer's disease (LOAD) is the leading cause of dementia. Recent large genome-wide association studies (GWAS) identified the first strongly supported LOAD susceptibility genes since the discovery of the involvement of APOE in the early 1990s. We have now exploited these GWAS datasets to uncover key LOAD pathophysiological processes. Methodology: We applied a recently developed tool for mining GWAS data for biologically meaningful information to a LOAD GWAS dataset. The principal findings were then tested in an independent GWAS dataset.

An introduction to biological nuclear magnetic resonance spectroscopy

ABSTRACT Nuclear magnetic resonance (NMR) spectroscopy is one of the most powerful analytical techniques available to biology. This review is an introduction to the potential of this method and is aimed at readers who have little or no experience in acquiring or analyzing NMR spectra. We focus on spectroscopic applications of the magnetic resonance effect, rather than imaging ones, and explain how various aspects of the NMR phenomenon make it a versatile tool with which to address a number of biological problems. Using detailed examples, we discuss the use of 1H NMR spectroscopy in mixture analysis and metabolomics, the use of 13C NMR spectroscopy in tracking isotopomers and determining the flux through metabolic pathways (‘fluxomics’) and the use of 31P NMR spectroscopy in monitoring ATP generation and intracellular pH homeotasis in vivo. Further examples demonstrate how NMR spectroscopy can be used to probe the physical environment of a cell by measuring diffusion and the tumbling rates of individual metabolites and how it can determine macromolecular structures by measuring the bonds and distances which separate individual atoms. We finish by outlining some of the key challenges which remain in NMR spectroscopy and we highlight how recent advances— such as increased magnet field strengths, cryogenic cooling, microprobes and hyperpolarisation—are opening new avenues for today’s biological NMR spectroscopists.

The effect of animals on human health and well-being

Substantial sums of money are invested annually in preventative medicine and therapeutic treatment for people with a wide range of physical and psychological health problems, sometimes to no avail. There is now mounting evidence to suggest that companion animals, such as dogs and cats, can enhance the health of their human owners and may thus contribute significantly to the health expenditure of our country. This paper explores the evidence that pets can contribute to human health and well-being. The article initially concentrates on the value of animals for short- and long-term physical health, before exploring the relationship between animals and psychological health, focusing on the ability of dogs, cats, and other species to aid the disabled and serve as a "therapist" to those in institutional settings. The paper also discusses the evidence for the ability of dogs to facilitate the diagnosis and treatment of specific chronic diseases, notably cancer, epilepsy, and diabetes. Mechanisms underlying the ability of animals to promote human health are discussed within a theoretical framework. Whereas the evidence for a direct causal association between human well-being and companion animals is not conclusive, the literature reviewed is largely supportive of the widely held, and long-standing, belief that "pets are good for us." © 2009 The Society for the Psychological Study of Social Issues.

Structure-Based Mutational Analysis of eIF4E in Relation to sbm1 Resistance to Pea Seed-Borne Mosaic Virus in Pea

Background: Pea encodes eukaryotic translation initiation factor eIF4E (eIF4E(S)), which supports the multiplication of Pea seed-borne mosaic virus (PSbMV). In common with hosts for other potyviruses, some pea lines contain a recessive allele (sbm1) encoding a mutant eIF4E (eIF4E(R)) that fails to interact functionally with the PSbMV avirulence protein, VPg, giving genetic resistance to infection.

Making Sense of Leaving Care: The contribution of Bridges Model of Transition to Understanding the Psycho-social process.

This paper is based on research into the transition of young people leaving public care in Romania. Using this specific country example, the paper aims to contribute to present understandings of the psycho-social transition of young people from care to independent living by introducing the use of Bridges (2002) to build on existing theories and literature. The research discussed involved mixed methods design and was implemented in three phases: semi-structured interviews with 34 care leavers, focus groups with 32 professionals, and a professional-service user working group. The overall findings confirmed that young people experience two different, but interconnected transitions - social and psychological - which take place at different paces. A number of theoretical perpectives are explored to make sense of this transition including attachment theory, focal theory and identity. In addition, a new model for understanding the complex process of transitions was adapted from Bridges’ (2002) to capture the clear complexity of transition which the findings demonstrated in terms of their psycho-social transition. The paper concludes with messages for leaving and after care services with an emphasis on managing the psycho-social transition from care to independent living.

Calbindin 2 (CALB2) Regulates 5-Fluorouracil Sensitivity in Colorectal Cancer by Modulating the Intrinsic Apoptotic Pathway

The role of the calcium binding protein, Calbindin 2 (CALB2), in regulating the response of colorectal cancer (CRC) cells to 5-Fluorouracil (5-FU) was investigated. Real-time RT-PCR and Western blot analysis revealed that CALB2 mRNA and protein expression were down-regulated in p53 wild-type and p53 null isogenic HCT116 CRC cell lines following 48 h and 72 h 5-FU treatment. Moreover, 5-FU-induced apoptosis was significantly reduced in HCT116 and LS174T CRC cell lines in which CALB2 expression had been silenced. Further investigation revealed that CALB2 translocated to the mitochondria following 5-FU treatment and that 5-FU-induced loss of mitochondrial membrane potential (Delta psi(m)) was abrogated in CALB2-silenced cells. Furthermore, CALB2 silencing decreased 5-FU-induced cytochrome c and smac release from the mitochondria and also decreased 5-FU-induced activation of caspases 9 and 3/7. Of note, co-silencing of XIAP overcame 5-FU resistance in CALB2-silenced cells. Collectively, these results suggest that following 5-FU treatment in CRC cell lines, CALB2 is involved in apoptosis induction through the intrinsic mitochondrial pathway. This indicates that CALB2 may be an important mediator of 5-FU-induced cell death. Moreover, down-regulation of CALB2 in response to 5-FU may represent an intrinsic mechanism of resistance to this anti-cancer drug.

Dysregulation in retinal para-inflammation and age-related retinal degeneration in CCL2 or CCR2 deficient mice

We have shown previously that a para-inflammatory response exists at the retinal/choroidal interface in the aging eye; and this response plays an important role in maintaining retinal homeostasis under chronic stress conditions. We hypothesized that dysregulation of the para-inflammatory response may result in an overt pro-inflammatory response inducing retinal degeneration. In this study, we examined this hypothesis in mice deficient in chemokine CCL2 or its cognate receptor CCR2. CCL2- or CCR2-deficient mice developed retinal degenerative changes with age, characterized as retinal pigment epithelial (RPE) cell and photoreceptor cell death. Retinal cell death was associated with significantly more subretinal microglial accumulation and increased complement activation. In addition, monocytes from CCL2- or CCR2-deficient mice had reduced capacity for phagocytosis and chemotaxis, expressed less IL-10 but more iNOS, IL-12 and TNF-a when compared to monocytes from WT mice. Complement activation at the site of RPE cell death resulted in C3b/C3d but not C5b-9 deposition, indicating only partial activation of the complement pathway. Our results suggest that altered monocyte functions may convert the protective para-inflammatory response into an overtly harmful inflammation at the retina/choroidal interface in CCL2- or CCR2-deficient mice, leading to RPE and photoreceptor degeneration. These data support a concept whereby a protective para-inflammatory response relies upon a normally functioning innate immune system. If the innate immune system is deficient chronic stress may tip the balance towards an overt inflammatory response causing cell/tissue damage.

Labor unions and Nash bargaining using coalition formation games

In this paper we examine the properties of stable coalitions under sequential and simultaneous bargaining by competing labor unions. We do this using the Nash bargaining solution and various notions of stability, namely, Nash, coalitional, contractual and core stability. (C) 2011 Elsevier B.V. All rights reserved,

Archiving Qualitative Data in the Context of a Society Coming out of Conflict: Some Lessons from Northern Ireland.

ARK (‘Access Research Knowledge’) was set up with a single goal: to make social science information on Northern Ireland available to the widest possible audience. The most well-known and widely used part of the ARK resource is CAIN (Conflict Archive on the INternet), which is one of the largest on-line collections of source material and information and about the Northern Ireland conflict. The compilation of CAIN's new Remembering: Victims, Survivors and Commemoration section raised issues related to the sensitivity of the material, as it feeds into the fundamental debate on the legacy of the Northern Ireland conflict. It also fundamentally raises the question to what extent archiving is a neutral or political activity and necessitates a discourse on responsibility and ethics among social researchers. Experiences from the establishment of the Northern Ireland Qualitative Archive (NIQA) shed light on future possibilities with regard to qualitative archives on the Northern Ireland conflict.

The Representation of the Female Body in the Multimedia Works of Regina José Galindo

The female body is central to the performance art, poetry and blog site interventions of Guatemalan Regina José Galindo. While Galindo is best known for her performance work, this article compares the hereto overlooked, distinctive and often shocking representations of the female body across her multimedia outputs. We first consider the ways in which, in all three media, Galindo presents an ‘excessive’, carnivalised, grotesque and abject female body. Second, we analyse representations of the female body that has been subjected to violence at a private and public level. In so doing, we show how Galindo not only contests hegemonic visions of gender and (national) identity but also challenges the viewer/reader to engage with, rather than look away from, the violence to which women are subjected in patriarchal society.