Investigating The Grey Field Slug

High-throughput sequencing was used to analyse cDNA generated from tissues of the grey field slug, Deroceras reticulatum, a significant invertebrate pest of agricultural and horticultural crops. Almost no sequence data is available for this organism. In this project, we performed de novo transcriptome sequencing to produce sequence dataset for the Deroceras reticulatum. A total of 132,597 and 161,419 sequencing reads between 50-600bp from the digestive gland and neural tissue were obtained through Roche 454 pyrosequencing. These reads were assembled into contiguous sequences and annotated using sequence homology search tools. Multiple sequence assemblies and annotation data was amalgamated into a biological database using BioSQL. Analysis of the dataset with predictions of probable protein function were made based on annotation data. InterPro (IPR) terms generated with InterProScan software were mapped to read counts and used to identify more frequently sequenced gene families. Digestive hydrolases were major transcripts in the digestive gland, with cysteine proteinases and cellulases being the most abundant functional classes. A Cathepsin L homologue is likely to be responsible for the proteinase activity of the digestive gland which was previously detected by biochemical analysis. Cathepsin L and several other predicted proteins were used to design RNAi experiments to assess potential for crop pest defence strategy. Further work on protein expression of a native tumour necrosis factor (TNF) ligand homologue was also conducted as an exemplar study

Establishing modular ligation through transglutaminase engineering

The ability to precisely direct proteins into ordered complexes is highly valuable for the synthetic biologist. Tools that can direct formation of covalent bonds facilitate particularly robust assemblies, which may enable new properties in areas such as optical control or programming of cell behaviour. Decades of research have produced an extensive range of tools for covalent conjugation of proteins both in vitro and in vivo, each with specific requirements and characteristics that determine the scope of their application. However, there is still a need for systems that can conjugate endogenous proteins in the extracellular space or precisely assemble endogenous proteins in vitro. Transglutaminases are a family of enzymes characterized by their ability to catalyse formation of a covalent ε-(γ-glutamyl) lysine isopeptide bond between protein substrates. Transglutaminase 2 (TG2) is a well-studied human transglutaminase with potential to provide unique functionality to the protein-protein coupling toolbox. The moderate specificity of TG2 for glutamine donors and low specificity for lysine donors presents the opportunity for covalent decoration of endogenous protein surfaces. This thesis aimed to explore the engineering of TG2 to enhance protein-protein conjugation. In Chapter 3 I demonstrated that TG2 is a substrate for itself and forms high molecular weight covalent multimers when activated. A truncated variant of TG2 (TG2465) retained transamidase activity while self-aggregation was abolished. I also demonstrated that TG2 interacts robustly with commonly used scaffold proteins and the cell surface. I determined kinetics of transamidase catalysis in different activation states and generated a variant with prolonged activity in oxidising conditions. I introduced glutamine- and lysine bearing peptide tags to model substrates and demonstrated that these tags impart TG2-reactivity. In Chapter 4 I investigated SnoopLigase, a protein-protein conjugation system derived from RrgA of Streptococcus pneumoniae. SnoopLigase requires very stringent reaction conditions, which limits application. I demonstrated considerable improvement in reaction efficiency for the SnoopLigase2 system in several common buffers and determined the contribution of each component to the overall increase in performance. I also found that SnoopLigase2 has a strong tolerance to temperature and pH, achieving conjugation in conditions that have not been demonstrated with any competing technologies. These findings open up new possibilities for protein-protein conjugation. In Chapter 5 I established protocols for efficient assembly of TG2:cargo conjugates using SnoopLigase2. I demonstrated TG2-mediated retention of TG2:cargo constructs to protein surfaces via non-covalent interactions. I generated a TG2:TGFα construct and achieved enhanced duration of TGFα activity due to TG2-mediated localisation at the cell surface. In this thesis, I have revealed properties of TG2 and SnoopLigase2 that offer interesting functionalities for the synthetic biologist and may be useful for molecular assembly in tissue engineering and bio-photonic applications

Democracy experts are divided on Lord Stevens’ proposals to reform police accountability

Last week the report of the Independent Police Commission, led by the former Metropolitan Police Commissioner John Stevens and commissioned by the Labour Party, was published. Its recommendations included the abolition of the post of Police and Crime Commissioner – introduced in 2012 to establish a new form of democratic oversight of the police – and the replacement of the Independent Police Complaints Commission. In this post we ask experts in democracy and police accountability to respond to the proposals and consider the implications of further reform

DOES PRE-EXERCISE STRETCHING AFFECT MUSCLE TENDON UNIT COMPLIANCE?

INTRODUCTION: Pre-exercise stretching is carried out by the majority of recreational and professional athletes, yet there is conflicting evidence with regard to the benefits of such programmes to either performance or injury risk reduction. It is unclear whether pre-exercise stretching can affect compliance, and at what level of the muscle-tendon unit (MTU) this effect may occur. Therefore the aim of this study was to determine whether an active preexercise stretching protocol increased the compliance of the triceps surae muscle-tendon complex and whether changes in compliance occurred in the muscle body or free tendon

Graduate police officers: releasing the potential for pre-employment university programmes for aspiring officers

The introduction of IPLDP (Initial Police Learning and Development Programme) provided Chief Constables with the opportunity to deliver initial police training through established police training approaches or involve other organisations. Since the 1970s there have been attempts by universities in the UK to engage with police services in the design and delivery of educational programmes for police officers. A variety of curriculums and new partnerships have evolved but there is little evidence of the contribution of these developments or whether these ‘new’ approaches differ significantly from traditional training regimes. There remains resistance from some quarters towards involving universities in the learning and development of police officers manifested by the Government's lead that situates the essential learning requirements of a police officer below higher education level. This article suggests that policing is at a crossroads and needs to decide now if it wants to be seen as a profession

Gambaran Jumlah Trombosit pada Pasien Kanker Serviks di RSU Dokter Soedarso Tahun 2011-2012

Latar Belakang: Rata Rata satu koma empat juta wanita di-seluruhdunia hidup dengan kanker serviks. Kanker serviks termasuk dari sepuluhbesar kanker penyebab kematian pada wanita di Indonesia. Hasilpenelitian terkini mengindikasikan adanya hubungan antara tingginyajumlah trombosit dengan prognosis yang buruk dari kanker sistemginekologi. Tujuan: Penelitian ini bertujuan mengetahui bagaimanagambaran jumlah trombosit pada pasien kanker serviks. Metodologi:Penelitian adalah penelitian deskriptif. Pengumpulan data dilakukan dibagian rekam medis RSU DOKTER SOEDARSO pada tanggal 25 Maretsampai 3 Mei 2013. Data dikumpulkan dari buku registrasi rawat inap danrawat jalan di poli obstetri dan ginekologi di RSU DOKTER SOEDARSO,sampel diambil secara consecutive sampling. Data dianalisis denganstatistik sederhana. Hasil: Ditemukan 39 sampel yang sesuai kriteriainklusi dan eksklusi dengan karakteristik sampel ; usia dengan 42 48tahun (35.9%), pekerjaan ibu rumah tangga (87.2%), dan stadium III/a(43.6%) adalah frekuensi yang tertinggi. Jumlah trombosit yang ditemukanpaling banyak adalah trombositosis (>400.000/ul) (64.1%). Rata-rata nilaitrombosit tertinggi yang ditemukan ada di stadium IV (592.000/ul).Kesimpulan: Distribusi jumlah trombosit terbanyak adalah trombositosis.Keadaan trombositosis mulai ditemukan pada stadium III/a. Nilai trombositmeningkat secara linier dengan stadium kanker serviks

Evaluation of a coastal acoustic buoy for cetacean detections, bearing accuracy and exclusion zone monitoring

The Maryland Department of Natural Resources and the Maryland Offshore Wind Development Fund at the Maryland Energy Administration cosponsored this work. This report was prepared as an account of work sponsored by an agency of the United States Government.There is strong socio-political support for offshore wind development in US territorial waters and construction is planned off several east coast states. Some of the planned development sites coincide with important habitat for critically endangered North Atlantic right whales. Both exclusion zones and passive acoustic monitoring are important tools for managing interactions between marine mammals and human activities. Understanding where animals are with respect to exclusion zones is important to avoid costly construction delays while minimizing the potential for negative impacts. Impact piling from construction of hundreds of offshore wind turbines likely require exclusion zones as large as 10 km. We have developed a three-hydrophone passive acoustic monitoring system that provides bearing information along with marine mammal detections to allow for informed management decisions in real-time. Multiple units form a monitoring system designed to determine whether marine mammal calls originate from inside or outside of an exclusion zone. In October 2021, we undertook a full system validation, with a focus on evaluating the detection range and bearing accuracy of the system with respect to right whale upcalls. Five units were deployed in Mid-Atlantic waters and we played more than 3500 simulated right whale upcalls at known locations to characterize the detection function and bearing accuracy of each unit. The modelled results of the detection function error were then used to compare the effectiveness of a bearing-based system to a single sensor that can only detect a signal but not ascertain directivity. Field trials indicated maximum detection ranges from 4-7.3 km depending on source and ambient noise levels. Simulations showed that incorporating bearing detections provide a substantial improvement in false alarm rates (6 to 12 times depending on number of units, placement and signal to noise conditions) for a small increase in the risk of missed detections inside of an exclusion zone (1%-3%). We show that the system can be used for monitoring exclusion zones and clearly highlight the value of including bearing estimation into exclusion zone monitoring plans while noting that placement and configuration of units should reflect anticipated ambient noise conditions.Publisher PDFPeer reviewe

Increased Wellbeing following Engagement in a Group Nature-Based Programme: The Green Gym Programme Delivered by the Conservation Volunteers

The wellbeing benefits of engaging in a nature-based programme, delivered by the Voluntary, Community and Social Enterprise sector, were examined in this study. Prior to attending The Conservation Volunteers’ Green Gym™, attendees (n = 892) completed demographics, health characteristics and the Warwick Edinburgh Mental Wellbeing Short-Form Scale. Attendees (n = 253, 28.4%) provided a measure on average 4.5 months later. There were significant increases in wellbeing after engaging in Green Gym, with the greatest increases in those who had the lowest starting levels of wellbeing. Wellbeing increases were sustained on average 8.5 months and 13 months later in those providing a follow up measure (n = 92, n = 40, respectively). Attendees who continued to engage in Green Gym but not provide follow up data (n = 318, 35.7%) tended to be more deprived, female and self-report a health condition. Attendees who did not continue to engage in Green Gym (n = 321, 36.0%) tended to be less deprived and younger. These findings provide evidence of the wellbeing benefits of community nature-based activities and social (‘green’) prescribing initiatives and indicate that Green Gym targets some groups most in need.</jats:p

Influenza Vaccine Effectiveness against Hospitalisation with Confirmed Influenza in the 2010-11 Seasons: A Test-negative Observational Study

Immunisation programs are designed to reduce serious morbidity and mortality from influenza, but most evidence supporting the effectiveness of this intervention has focused on disease in the community or in primary care settings. We aimed to examine the effectiveness of influenza vaccination against hospitalisation with confirmed influenza. We compared influenza vaccination status in patients hospitalised with PCR-confirmed influenza with patients hospitalised with influenza-negative respiratory infections in an Australian sentinel surveillance system. Vaccine effectiveness was estimated from the odds ratio of vaccination in cases and controls. We performed both simple multivariate regression and a stratified analysis based on propensity score of vaccination. Vaccination status was ascertained in 333 of 598 patients with confirmed influenza and 785 of 1384 test-negative patients. Overall estimated crude vaccine effectiveness was 57% (41%, 68%). After adjusting for age, chronic comorbidities and pregnancy status, the estimated vaccine effectiveness was 37% (95% CI: 12%, 55%). In an analysis accounting for a propensity score for vaccination, the estimated vaccine effectiveness was 48.3% (95% CI: 30.0, 61.8%). Influenza vaccination was moderately protective against hospitalisation with influenza in the 2010 and 2011 seasons

Commercial ship versus whale watch boat noise: relative effects on Southern Resident killer whales

Underwater noise may be impacting the population recovery of critically endangered Southern Resident Killer Whales (SRKW). This study used an SRKW-Noise exposure simulation model to compare noise effects from large (AIS-enabled) commercial vessels with whale watch boats during summer (May-September) within their principal Salish Sea habitat range use. It predicted moderate or low behavioural responses (BRs) using SRKW-specific dose-response relationships and, if no BRs were triggered, the extent of residual high frequency echolocation click masking. BRs were considered to result in lost foraging due to switches in behaviour or via strong masking effects. The Monte-Carlo simulation used a fine-scale acoustic model to predict broadband sound pressure levels (SPL, BR analysis) and power spectral density (PSD) at 50 kHz (click masking analysis) for AIS-enabled commercial vessels. To derive equivalent data from whale watch boats, we combined data from Holt et al. (2009), SoundWatch and Beam Reach Sustainable School. SRKW habitat used a 10-year synthesis of effort-correct observer sightings. Overall, noise from AIS-enabled vessels was estimated to contribute 93% of overall BR-related potential lost foraging time, with whale watch boats contributing the remaining 7%, despite mean estimates of 6.1hr of boats with each whale per day. Lower SPLs of slow moving boats had low probabilities of exceeding BR thresholds, while large commercial vessel often exceed these thresholds. Lost foraging time per whale was estimated as a median 3.2hr per day when whales were present (13.4% of day). Echolocation click masking effects accumulated an additional 1.7hr of lost foraging time and was strongly dominated by noise predicted from slow (2.5-8 knot) whale watch boats, noting high model uncertainty due to PSD, speed and proximity assumptions. Overall, lost foraging time totaled 20.3% of each whale day (4.9hr), with ~2/3 due to AIS-enabled commercial vessels, highlighting mitigation measures for both vessel types should be considered