Equal but Different: Gender Discourses in the Social Relations of Irish Peacekeepers & Possibilities for Transformation

Equal but Different: Discourses in the Social Relations of Irish Peacekeepers & Possibilities for Transformation Motivated by the lack of action and transformation of gender relations within peacekeeping since the adoption of UNSCR 1325 in 2000, this study explores the gendering processes within the Irish Defence Forces that position women and men in particular roles informally and which act to support or inhibit women’s access to peacekeeping missions. Through discourse analysis this study reveals ‘equal but different’ as the dominant discourse on gender relations within the Irish Defence Forces and uses this discourse as a lens through which to assess the overarching question: ‘How does the “equal but different” discourse distribute power in different contexts and what impact does that have on women’s inclusion in PSOs?’ By drawing out the empirical data from the accounts of 28 women and men participants the findings reveal what women bring to a mission; inhibitors to their participation in missions; and transformative possibilities. The study’s major contribution is that it reveals multiple contradictory discourses depending on the context. Of particular importance to the feminist agenda is this study’s new empirical data on Irish peacekeepers and the development of critical alternative discourses on gender as a result of women’s presence in missions. These alternative discourses have the potential to transform gender relations by positioning women and men in the ‘third space’ which holds ‘equal, ‘different’ and ‘multiplicity’ of subjectivities simultaneously. This ‘third space’ creates a bridge between the liberal and critical feminist debates on women’s participation in peacekeeping, through its development of a new concept ‘add women and transform’. The ‘add women and transform’ concept is borne out of the empirical findings revealing how the presence of women in the military is leading to the creation of new critical discourses, and although they are muted, they have the potential to challenge unequal power dynamics within the military if they are supported by gender mainstreaming policies and a shift in peacekeeping practices

Molecular and Genetic Analysis of the vha16 Gene in Drosophila melanogaster

Vacuolar H+-ATPases (V-ATPases) are multi-subunit enzymes which pump protons across cellular membranes via hydrolysis of ATP. They are related, both in structure and function, to FI-FQ ATP synthases and much about the V-ATPase mode of action has been inferred from FI-FQ studies. V-ATPases have been isolated from a broad phylogenetic range of eukaryotes, from single celled organisms to higher plants and animals. In these higher organisms they can play both endomembrane and plasma membrane roles in a large array of cell types. Several genes encoding subunits of the V-ATPase holoenzyme have been cloned from the model genetic organism Drosophila melanogaster. One of the subunits cloned from D. melanogaster, vhal6, encodes the 16 kDa pore forming protein, through which the hydrogen ions pass. Firstly, this project focuses on this proteolipid gene in Drosophila melanogaster, describing the isolation and characterisation of a lethal P-element insert within the originally described gene, and discussing how the insertion of this element disrupts the normal function of the gene. Also discussed is how this P-element can be used to show the spatial and temporal expression of the gene in vivo. Secondly it describes the isolation of four new homologues of this gene in D. melanogaster. It show's their sequence relationship to the original Drosophila protein, and the protein isolated from other organisms and discusses if they are expressed and whether they would function as V-ATPase pores. This study also describes the attempted rescue of the lethal P-element phenotype with a range of proteolipid sequences cloned from Drosophila and also proteolipids of other organisms. Lastly, this project describes the isolation of two other Drosophila V-ATPase membrane associated subunits, one homologous to PPA1 in yeast and the other homologous to M9.7 in Manduca sexta

Structure and mechanism of the type I-G CRISPR effector

Funding: This work was supported by the Biotechnology and Biological Sciences Research Council (REF: BB/S000313/1 to MFW), Medical Research Council (REF: MR/S021647/1 to RS) and the China Scholarship Council (REF: 202008060345 to QS).Type I CRISPR systems are the most common CRISPR type found in bacteria. They use a multisubunit effector, guided by crRNA, to detect and bind dsDNA targets, forming an R-loop and recruiting the Cas3 enzyme to facilitate target DNA destruction, thus providing immunity against mobile genetic elements. Subtypes have been classified into families A-G, with type I-G being the least well understood. Here, we report the composition, structure and function of the type I-G Cascade CRISPR effector from Thioalkalivibrio sulfidiphilus, revealing key new molecular details. The unique Csb2 subunit processes pre-crRNA, remaining bound to the 3′ end of the mature crRNA, and seven Cas7 subunits form the backbone of the effector. Cas3 associates stably with the effector complex via the Cas8g subunit and is important for target DNA recognition. Structural analysis by cryo-Electron Microscopy reveals a strikingly curved backbone conformation with Cas8g spanning the belly of the structure. These biochemical and structural insights shed new light on the diversity of type I systems and open the way to applications in genome engineering.Publisher PDFPeer reviewe

Prespacer processing and specific integration in a Type I-A CRISPR system

This work was supported by a grant from the Biotechnology and Biological Sciences Research Council (REF: BB/M021017/1 to MFW).The CRISPR–Cas system for prokaryotic adaptive immunity provides RNA-mediated protection from viruses and mobile genetic elements. Adaptation is dependent on the Cas1 and Cas2 proteins along with varying accessory proteins. Here we analyse the process in Sulfolobus solfataricus, showing that while Cas1 and Cas2 catalyze spacer integration in vitro, host factors are required for specificity. Specific integration also requires at least 400 bp of the leader sequence, and is dependent on the presence of hydrolysable ATP, suggestive of an active process that may involve DNA remodelling. Specific spacer integration is associated with processing of prespacer 3′ ends in a PAM-dependent manner. This is reflected in PAM-dependent processing of prespacer 3′ ends in vitro in the presence of cell lysate or the Cas4 nuclease, in a reaction consistent with PAM-directed binding and protection of prespacer DNA. These results highlight the diverse interplay between CRISPR–Cas elements and host proteins across CRISPR types.Publisher PDFPeer reviewe

The Exchange of the Actin-bound Nucleotide

Radioactive adenine is readily taken up by muscle and is incorporated into nucleotides as an intact unit. The rate of uptake is independent of the duration of incubation and increases with temperature. About 1/6 of the actin-bound ADP is rapidly exchangeable with free nucleotide in resting muscle. In isometric contraction there is a slight loss of tritium label from the 2 position of the actin-bound ADP. In loaded contracture there is a slight exchange of the actin-bound nucleotide over and above the resting exchange. The extent of this additional exchange seems to parallel the peak power output of the muscle. For a muscle contracting at maximum power, the probability that a bound ADP will exchange during a single interaction with myosin is about 0.0005. Experiments in vitro show that, at high temperatures, the F-actin-bound ADP is labile and that the ATPase of the actin is dependent on the preparation of the protein. It is possible to calculate the length (along the thin filament) between the points at which any one cross bridge makes successive attachments. For the purposes of this calculation it is only necessary to know certain mechanical, energetic and geometric properties of muscle; no particular model of muscular contraction need be assumed. The length in question is about 58 nm, the length of the half turn of the thin filament.<p

The Exchange of the Actin-bound Nucleotide

Radioactive adenine is readily taken up by muscle and is incorporated into nucleotides as an intact unit. The rate of uptake is independent of the duration of incubation and increases with temperature. About 1/6 of the actin-bound ADP is rapidly exchangeable with free nucleotide in resting muscle. In isometric contraction there is a slight loss of tritium label from the 2 position of the actin-bound ADP. In loaded contracture there is a slight exchange of the actin-bound nucleotide over and above the resting exchange. The extent of this additional exchange seems to parallel the peak power output of the muscle. For a muscle contracting at maximum power, the probability that a bound ADP will exchange during a single interaction with myosin is about 0.0005. Experiments in vitro show that, at high temperatures, the F-actin-bound ADP is labile and that the ATPase of the actin is dependent on the preparation of the protein. It is possible to calculate the length (along the thin filament) between the points at which any one cross bridge makes successive attachments. For the purposes of this calculation it is only necessary to know certain mechanical, energetic and geometric properties of muscle; no particular model of muscular contraction need be assumed. The length in question is about 58 nm, the length of the half turn of the thin filament.<p

Fuse to defuse : a self-limiting ribonuclease-ring nuclease fusion for type III CRISPR defence

Funding: Biotechnology and Biological Sciences Research Council [REF BB/S000313/1 to M.F.W.]; Funding for open access charge: RCUK block grant.Type III CRISPR systems synthesise cyclic oligoadenylate (cOA) second messengers in response to viral infection of bacteria and archaea, potentiating an immune response by binding and activating ancillary effector nucleases such as Csx1. As these effectors are not specific for invading nucleic acids, a prolonged activation can result in cell dormancy or death. Some archaeal species encode a specialised ring nuclease enzyme (Crn1) to degrade cyclic tetra-adenylate (cA4) and deactivate the ancillary nucleases. Some archaeal viruses and bacteriophage encode a potent ring nuclease anti-CRISPR, AcrIII-1, to rapidly degrade cA4 and neutralise immunity. Homologues of this enzyme (named Crn2) exist in type III CRISPR systems but are uncharacterised. Here we describe an unusual fusion between cA4-activated CRISPR ribonuclease (Csx1) and a cA4-degrading ring nuclease (Crn2) from Marinitoga piezophila. The protein has two binding sites that compete for the cA4 ligand, a canonical cA4-activated ribonuclease activity in the Csx1 domain and a potent cA4 ring nuclease activity in the C-terminal Crn2 domain. The cA4 binding affinities and activities of the two constituent enzymes in the fusion protein may have evolved to ensure a robust but time-limited cOA-activated ribonuclease activity that is finely tuned to cA4 levels as a second messenger of infection.Publisher PDFPeer reviewe

Competition for Pollination Between an Invasive Species (Purple Loosestrife) and a Native Congener

Invasive species are frequently regarded as Superlative competitors that can vegetatively crowd Out natives, but little is known about whether invasives call compete for pollination services with native plants. We hypothesized that, when the showy invasive species Lythrum salicaria (purple loosestrife) was present, pollinator visitation and seed set would be reduced in a native congener, L. alatum (winged loosestrife). To test this hypothesis, we constructed mixed and monospecific plots of the two species. Over two years of study, we found that L. salicaria significantly reduced both pollinator visitation and seed set in L. alatum. Furthermore, pollinators moved frequently between the two plant species, which may cause heterospecific pollen transfer. Thus, reductions in both pollen quantity and pollen quality may reduce L. alatum seed set. If similar patterns occur in the field, invasive plants may be an even greater threat to natives than previously thought

Structure of the Saccharolobus solfataricus type III-D CRISPR effector

We acknowledge funding from BBSRC BB/J005673/1 project grant to LS and MFW and ERC funding to MFW (grant ref 101018608). DK was funded by a Darwin Trust of Edinburgh grant. We acknowledge Diamond Light Source for access and support of the cryo-EM facilities at the UK's national Electron Bio-imaging Centre (eBIC) under proposal EM16637-14, funded by the Wellcome Trust, MRC and BBRSC. The Scottish Centre for Macromolecular Imaging (SCMI) is funded by the MRC (MC_PC_17135) and SFC (H17007).CRISPR-Cas is a prokaryotic adaptive immune system, classified into six different types, each characterised by a signature protein. Type III systems, classified based on the presence of a Cas10 subunit, are rather diverse multi- subunit assemblies with a range of enzymatic activities and downstream ancillary effectors. The broad array of current biotechnological CRISPR applications is mainly based on proteins classified as Type II, however recent developments established the feasibility and efficacy of multi-protein Type III CRISPR-Cas effector complexes as RNA-targeting tools in eukaryotes. The crenarchaeon Saccharolobus solfataricus has two type III system subtypes (III- B and III-D). Here, we report the cryo-EM structure of the Csm Type III-D complex from S. solfataricus (SsoCsm), which uses CRISPR RNA to bind target RNA molecules, activating the Cas10 subunit for antiviral defence. The structure reveals the complex organisation, subunit/subunit connectivity and protein/guide RNA interactions of the SsoCsm complex, one of the largest CRISPR effectors known.Publisher PDFPeer reviewe

TennisSense: a multi-sensory approach to performance analysis in tennis

The TennisSense Project, that is run in collaboration with Tennis Ireland, aims to create the infrastructure required to digitally capture physical, tactical and physiological data from tennis players in order to assist in their coaching and improved performance. This study examined the potential for using Wireless Inertial Monitoring Units (WIMU) to model the biomechanical aspects of the tennis stroke and for developing coaching tools that utilise this information. There is significant evidence in the current literature that the ability to accurately capture and model the accelerations, angular velocities and orientations involved in the tennis stroke could facilitate a major step forward in the application of biomechanics to tennis coachin