Attorneys\u27 Fees and the Conflict Between Rule 68 and the Clean Water Act\u27s Citizen Suit Provision

Environmental “citizen suit” statutes provide incentives for citizens to bring enforcement actions by awarding successful plaintiffs reasonable attorneys’ fees. Defendants have attempted to use Federal Rule of Civil Procedure 68 to block a successful plaintiff’s recovery of attorneys’ fees. Under Rule 68, defendants may offer to allow a judgment to be issued against them for a fixed dollar amount. Plaintiffs may either accept this judgment offer or proceed to trial. If plaintiffs proceed to trial, however, they must receive a judgment more favorable than the offer or pay the defendants’ litigation costs. Defendants argue that the word “costs” as used in Rule 68 applies to attorneys’ fees in addition to other litigation costs. If so, the use of Rule 68 can have a great influence on the economics of citizen suit litigation. This Note explores whether or not Rule 68 should be read to apply to attorneys’ fees in citizen suits under the Clean Water Act and other environmental statutes

Gold Particle Analyser: Detection and quantitative assessment of electron microscopy gold probes

Gold particle probes are an essential electron microscopy tool to examine protein localisation, as well as protein trafficking. They can be introduced into living cells when conjugated to a protein that is endocytosed or to an antibody against a cell surface protein. Alternatively, gold particles can be introduced into fixed cells or tissue when conjugated to antibodies, immunoglobulin binding molecules or chemical probes applied to permeabilised samples or electron microscopy sections. Colloidal gold particles that have not been enlarged through chemical (gold or silver) enhancement are typically spherical and can be prepared in a range of specific sizes, allowing multiple proteins to be localised within a single sample. The typically homogeneous shape and size of the colloidal gold makes them ideal for computer assisted detection and analysis. Here we demonstrate a program developed to automatically identify two sizes of gold particle and perform a range of analyses that includes (i) distribution and cluster analysis; (ii) selection and analysis of gold particles allocated close to or either side of a membrane; (iii) measurement of organelle size; (iv) estimation of the number of gold particles within an aggregate and (v) the detection of chemically enhanced irregular sized and shaped gold particles. We show this easy-to-use program can greatly assist electron microscopists, to reliably and efficiently analyse gold particles within their images

Observation of Classroom Performance Using Therapy Balls as a Substitute for Chairs in Elementary School Children

Many classrooms are beginning to substitute standard chairs with therapy balls, which help to improve students’ focus and classroom performance, according to teacher and student reports. Researchers conducted an observational study in a classroom at a local elementary school that implemented therapy balls. For each hour-long observation, three independent researchers marked seated behaviors of students (N=19). The first observation was conducted when students were using standard chairs, and two more were conducted when students used therapy balls during classroom activities. Researchers observed the behaviors of students on the given seating device, including academic task (on/off task), effort level, attitude, interactions, seated behavior (bouncing/rocking/stationary/other), and intensity level of participation in classroom activities. Analysis of the collected data demonstrates that 50% of observations when students were seated on standard chairs indicated on task behavior, while 85% of observations when students were seated on therapy balls were recorded as on task behavior. In addition, movement while seated increased on the therapy balls (35% of observations indicated stationary seated behavior) compared to the standard chairs (47% observations indicated stationary seated behavior). The increased rocking and bouncing seated behavior on the therapy balls suggests that the seating device helps to engage the vestibular system. In addition, stationary behavior on therapy balls was often classified as proprioceptive. The results from the observational study will allow researchers to further investigate whether or not engaging the vestibular system can help to improve performance on functional school skills

Cohabitation and the law: myths, money and the media

This chapter version has been deposited with the kind permission of Sage. The definitive version of this chapter was published by Sage in the 24th British Social Attitudes Report 2008, by Alison Park, John Curtice, Katarina Thomson, Miranda Phillips, Mark Johnson, Elizabeth Clery. ISBN: 9781412947749. Available at: http://www.uk.sagepub.com/books/Book231969The 2000 British Social Attitudes survey confirmed a growing social acceptance of heterosexual cohabitation as a partnering and parenting choice and identified strong public support for reform of cohabitation law. It also established the existence of a ‘common law marriage’ myth whereby the majority of the public, and cohabitants in particular, falsely believe that cohabiting couples who have lived together for some time have the same legal rights as married couples. Even among those cohabitants who were aware of their vulnerable legal position, it was found that very few had taken appropriate steps to gain or provide legal protection despite, as we found in subsequent research, often having good intentions to do so. These findings prompted widespread media interest and government concern. This led the Department for Constitutional Affairs (now the Ministry of Justice) to fund their 2004 Living Together Campaign, aimed at advising cohabitants about their legal rights and indicating practical steps they could take to gain marriage-like protection where possible. Subsequently, in 2005, with Scotland having already decided to reform the law relating to cohabiting couples (see Family Law (Scotland) Act 2006), the government decided to refer the issue of whether cohabitation law should be reformed to the Law Commission for England and Wales, a decision which sparked further media interest. The attention paid to cohabitation over the last few years makes it worth revisiting the subject, to see whether, and how, behaviour and attitudes are changing. So our first aim in this chapter is to examine the evolving prevalence and role of cohabitation as a relationship form in British society. Our second aim is to establish whether attitudes to cohabitation have changed, and whether legal knowledge and actions have increased following a period of sustained government and media focus. Finally, we consider public beliefs about cohabitants’ legal rights and financial practices. This latter issue is vital, as the courts may now take into account how cohabiting couples manage their money in deciding appropriate legal remedies on separation. Some of our key findings have already informed the deliberations of the Law Commission, due to report to Parliament in 2007, but this is the first full account of our research

AMP-activated protein kinase is a key regulator of acute neurovascular permeability

Many neuronal and retinal disorders are associated with pathological hyperpermeability of the microvasculature. We have used explants of rodent retinae to study acute neurovascular permeability and signal transduction and the role of AMP-activated protein kinase (AMPK). Following stimulation with either vascular endothelial growth factor (VEGF-A) or bradykinin (BK), AMPK was rapidly and strongly phosphorylated and acted as a key mediator of permeability downstream of Ca2+ Accordingly, AMPK agonists potently induced acute retinal vascular leakage. AMPK activation led to phosphorylation of endothelial nitric oxide synthase (eNOS), which in turn increased VE-cadherin phosphorylation on Y685. In parallel, AMPK also mediated phosphorylation of p38 MAP kinase and HSP27, indicating that it regulated paracellular junctions and cellular contractility, both previously associated with endothelial permeability. Endothelial AMPK provided a missing link in neurovascular permeability, connecting Ca2+ transients to the activation of eNOS and p38, irrespective of the permeability-inducing factor used. Collectively, we find that, due to its compatibility with small molecule antagonists/agonists and siRNA, the ex-vivo retina model constitutes a reliable tool to identify and study regulators and mechanism of acute neurovascular permeability

Chronically shortened rod outer segments accompany photoreceptor cell death in Choroideremia

X-linked choroideremia (CHM) is a disease characterized by gradual retinal degeneration caused by loss of the Rab Escort Protein, REP1. Despite partial compensation by REP2 the disease is characterized by prenylation defects in multiple members of the Rab protein family that are master regulators of membrane traffic. Remarkably, the eye is the only organ affected in CHM patients, possibly because of the huge membrane traffic burden of the post mitotic photoreceptors, which synthesise outer segments, and the adjacent retinal pigment epithelium that degrades the spent portions each day. In this study, we aimed to identify defects in membrane traffic that might lead to photoreceptor cell death in CHM. In a heterozygous null female mouse model of CHM (Chmnull/WT), degeneration of the photoreceptor layer was clearly evident from increased numbers of TUNEL positive cells compared to age matched controls, small numbers of cells exhibiting signs of mitochondrial stress and greatly increased microglial infiltration. However, most rod photoreceptors exhibited remarkably normal morphology with well-formed outer segments and no discernible accumulation of transport vesicles in the inner segment. The major evidence of membrane trafficking defects was a shortening of rod outer segments that was evident at 2 months of age but remained constant over the period during which the cells die. A decrease in rhodopsin density found in the outer segment may underlie the outer segment shortening but does not lead to rhodopsin accumulation in the inner segment. Our data argue against defects in rhodopsin transport or outer segment renewal as triggers of cell death in CHM.publishersversionpublishe

Changes in Mitochondrial Size and Morphology in the RPE and Photoreceptors of the Developing and Ageing Zebrafish

Mitochondria are essential adenosine triphosphate (ATP)-generating cellular organelles. In the retina, they are highly numerous in the photoreceptors and retinal pigment epithelium (RPE) due to their high energetic requirements. Fission and fusion of the mitochondria within these cells allow them to adapt to changing demands over the lifespan of the organism. Using transmission electron microscopy, we examined the mitochondrial ultrastructure of zebrafish photoreceptors and RPE from 5 days post fertilisation (dpf) through to late adulthood (3 years). Notably, mitochondria in the youngest animals were large and irregular shaped with a loose cristae architecture, but by 8 dpf they had reduced in size and expanded in number with more defined cristae. Investigation of temporal gene expression of several mitochondrial-related markers indicated fission as the dominant mechanism contributing to the changes observed over time. This is likely to be due to continued mitochondrial stress resulting from the oxidative environment of the retina and prolonged light exposure. We have characterised retinal mitochondrial ageing in a key vertebrate model organism, that provides a basis for future studies of retinal diseases that are linked to mitochondrial dysfunction

Symmetric arrangement of mitochondria:plasma membrane contacts between adjacent photoreceptor cells regulated by Opa1

Mitochondria are known to play an essential role in photoreceptor function and survival that enables normal vision. Within photoreceptors, mitochondria are elongated and extend most of the inner-segment length, where they supply energy for protein synthesis and the phototransduction machinery in the outer segment, as well as acting as a calcium store. Here, we examined the arrangement of the mitochondria within the inner segment in detail using three-dimensional (3D) electron microscopy techniques and show they are tethered to the plasma membrane in a highly specialized arrangement. Remarkably, mitochondria and their cristae openings align with those of neighboring inner segments. The pathway by which photoreceptors meet their high energy demands is not fully understood. We propose this to be a mechanism to share metabolites and assist in maintaining homeostasis across the photoreceptor cell layer. In the extracellular space between photoreceptors, Müller glial processes were identified. Due to the often close proximity to the inner-segment mitochondria, they may, too, play a role in the inner-segment mitochondrial arrangement as well as metabolite shuttling. OPA1 is an important factor in mitochondrial homeostasis, including cristae remodeling; therefore, we examined the photoreceptors of a heterozygous Opa1 knockout mouse model. The cristae structure in the Opa1+/− photoreceptors was not greatly affected, but the mitochondria were enlarged and had reduced alignment to neighboring inner-segment mitochondria. This indicates the importance of key regulators in maintaining this specialized photoreceptor mitochondrial arrangement