Search

Abstract The order Orthoptera includes insects such as grasshoppers, katydids, and crickets, among which there are important species for ecosystem stability and pollination, as well as research organisms in different fields such as neurobiology, ecology, and evolution. Crickets, with more than 2,400 described species, are emerging as novel model research organisms, for their diversity, worldwide distribution, regeneration capacity, and their characteristic acoustic communication. Here we report the assembly and annotation of the first New World cricket, that of Gryllus longicercus Weissman & Gray 2019. The genome assembly, generated by combining 44.54 Gb of long reads from PacBio and 120.44 Gb of short Illumina reads, has a length of 1.85 Gb. The genome annotation yielded 19,715 transcripts from 14,789 gene models.

This paper considers housing activity transmission across space using local indicators of spatial association. Activity does not diffuse like a ripple across England and Wales. Elevated activity appears to be associated with a south east-north west divide. The south east grows more quickly than the north west for a period of time, after which there is a switch in rank order. It is argued that changes in activity is associated with changing credit conditions to which the south east is more sensitive. Furthermore, it is proposed that activity features a type of participant induced on to the market by favourable price changes and credit conditions. It is argued that the reduced activity associated with 'missing mortgage movers' post-2007 reflects the lack of mortgage mover participation, which could have negative consequences for the macroeconomy.

A high-throughput screening pipeline tests the feasibility of targeting the sporozoite stage of P. falciparum through chemical interventions and identifies gramicidin as a potential sporontocidal agent.

B cells are key pathogenic drivers of chronic inflammation in rheumatoid arthritis (RA). There is limited understanding of the relationship between synovial B cell subsets and pathogenic antibody secreting cells (ASCs). This knowledge is crucial for the development of more targeted B-cell depleting therapies. While CD11c+ double-negative 2 (DN2) B cells have been suggested as an ASC precursor in lupus, to date there is no proven link between the two subsets in RA. We have used both single-cell gene expression and BCR sequencing to study synovial B cells from patients with established RA, in addition to flow cytometry of circulating B cells. To better understand the differentiation patterns within the diseased tissue, a combination of RNA-based trajectory inference and clonal lineage analysis of BCR relationships were used. Both forms of analysis indicated that DN2 B cells serve as a major precursors to synovial ASCs. This study advances our understanding of B cells in RA and reveals the origin of pathogenic ASCs in the RA synovium. Given the significant role of DN2 B cells as a progenitor to pathogenic B cells in RA, it is important to conduct additional research to investigate the origins of DN2 B cells in RA and explore their potential as therapeutic targets in place of the less specific pan-B cells depletion therapies currently in use.

It is likely that Autonomous Vehicles will have significant social, cultural, spatial and environmental implications and that the interaction between humans, automated vehicles and physical environment will provide an array of challenges. This paper aims to explore the use of innovative visualisation approaches, to foster discussion on possible scenarios involving AVs. It is argued that such an approach might be used to help conceptualise human experiences with the potential to enhance understanding of the complex human-machine associations. Presenting journeys from different perspectives and reconceptualising the context through the eyes of AVs emphasized the nuances of experience between the machines, urban space and human bodies. Unexpected user-technology interactions will emerge as humans are not always passive followers and can be apprehensive when it comes to accepting such a novel technology as self-driving vehicles. The focus applied in the methodology and data capture was on inclusivity of data, showing not only movement but also noise and human experience of a space. The integration of AVs on public roads will rely on technical innovation to ensure that vehicles can operate safely yet, the study of the perceptual and ethical effects of technology and potential influences on society via engaging the public will help to manage expectations and create platforms for mutual learning.

An ecosystem-based management approach (EBM) is suggested as one solution to help to tackle environmental challenges facing worldwide farming systems whilst ensuring socio-economic demands are met. Despite its usefulness, the application of this approach at the farm-scale presents several implementation problems, including the difficulty of (a) incorporating the concept of ecosystem services (ES) into agricultural land use decision-making and (b) involving the farmer in the planning process. This study aims to propose a solution to overcome these challenges by utilising a geodesign framework and EBM approach to plan and design a sustainable multifunctional agricultural landscape at the farm scale. We demonstrate how the proposed approach can be applied to plan and design multifunctional agricultural landscapes that offer improved sustainability, using a New Zealand hill country farm as a case study. A geodesign framework is employed to generate future land use and management scenarios for the study area, visualize changes, and assess the impacts of future land use on landscape multifunctionality and the provision of associated ES and economic outcomes. In this framework, collaboration with the farmer was carried out to obtain farm information and co-design the farmed landscapes. The results from our study demonstrate that farmed landscapes where multiple land use/ land cover types co-exist can provide a wide range of ES and therefore, meet both economic and environmental demands. The assessment of impacts for different land use change scenarios demonstrates that land use change towards increasing landscape diversity and complexity is a key to achieving more sustainable multifunctional farmed landscapes. The integration of EBM and geodesign, is a transdisciplinary approach that can help farmers target land use and management decisions by considering the major ES that are, and could be, provided by the landscapes in which these farm systems are situated, therefore maximising the potential for beneficial outcomes.

Dopamine agonists (DAs) have demonstrated efficacy for the treatment of Parkinson’s disease (PD) but are limited by adverse effects (AEs). DAs can vary considerably in their receptor subtype selectivity and affinity, chemical composition, receptor occupancy, and intrinsic activity on the receptor. Most currently approved DAs for PD treatment primarily target D2/D3 (D2-like) dopamine receptors. However, selective activation of D1/D5 (D1-like) dopamine receptors may enable robust activation of motor function while avoiding AEs related to D2/D3 receptor agonism. Full D1/D5 receptor-selective agonists have been explored in small, early-phase clinical studies, and although their efficacy for motor symptoms was robust, challenges with pharmacokinetics, bioavailability, cardiovascular AEs, and dyskinesia rates similar to levodopa prevented clinical advancement. Generally, repeated dopaminergic stimulation with full DAs is associated with frontostriatal dysfunction and sensitization that may induce plastic changes in the motor system, and neuroadaptations that produce long-term motor and nonmotor complications, respectively. Recent preclinical and clinical studies suggest that a D1/D5 receptor-selective partial agonist may hold promise for providing sustained, predictable, and robust motor control, while reducing risk for motor complications (e.g., levodopa-induced dyskinesia) and nonmotor AEs (e.g., impulse control disorders and excessive daytime sleepiness). Clinical trials are ongoing to evaluate this hypothesis. The potential emerging availability of novel dopamine receptor agonists with selective dopamine receptor pharmacology suggests that the older terminology “dopamine agonist” may need revision to distinguish older-generation D2/D3–selective agonists from D1/D5-selective agonists with distinct efficacy and tolerability characteristics.

Responsible artificial intelligence guidelines ask engineers to consider how their systems might harm. However, contemporary artificial intelligence systems are built by composing many preexisting software modules that pass through many hands before becoming a finished product or service. How does this shape responsible artificial intelligence practice? In interviews with 27 artificial intelligence engineers across industry, open source, and academia, our participants often did not see the questions posed in responsible artificial intelligence guidelines to be within their agency, capability, or responsibility to address. We use Suchman's “located accountability” to show how responsible artificial intelligence labor is currently organized and to explore how it could be done differently. We identify cross-cutting social logics, like modularizability, scale, reputation, and customer orientation, that organize which responsible artificial intelligence actions do take place and which are relegated to low status staff or believed to be the work of the next or previous person in the imagined “supply chain.” We argue that current responsible artificial intelligence interventions, like ethics checklists and guidelines that assume panoptical knowledge and control over systems, could be improved by taking a located accountability approach, recognizing where relations and obligations might intertwine inside and outside of this supply chain.

Abstract Radioactive iodine (RAI) is safe and effective in most patients with hyperthyroidism but not all individuals are cured by the first dose, and most develop post-RAI hypothyroidism. Postoperative RAI therapy for remnant ablation is successful in 80–90% of thyroid cancer patients and sometimes induces remission of nonresectable cervical and/or distant metastatic disease but the effective tumor dose is usually not precisely known and must be moderated to avoid short- and long-term adverse effects on other tissues. The Collar Therapy Indicator (COTI) is a radiation detection device embedded in a cloth collar secured around the patient’s neck and connected to a recording and data transmission box. In previously published experience, the data can be collected at multiple time points, reflecting local cervical RAI exposure and correlating well with conventional methods. We evaluated the real-time uptake of RAI in patients with hyperthyroid Graves’ disease and thyroid cancer. We performed a pilot feasibility prospective study. Data were analyzed using R© (version 4.0.3, The R Foundation for Statistical Computing, 2020), and Python (version 3.6, Matplotlib version 3.0.3). The COTI was able to provide a quantitative temporal pattern of uptake within the thyroid in persons with Graves’ disease and lateralized the remnant tissue in persons with thyroid cancer. The study has demonstrated that the portable collar radiation detection device outside of a healthcare facility is accurate and feasible for use after administration of RAI for diagnostic studies and therapy to provide a complete collection of fractional target radioactivity data compared to that traditionally acquired with clinic-based measurements at one or two time-points. Clinical Trials Registration NCT03517579, DOR 5/7/2018.

Using the Christiano-Fitzgerald filter to extract business and medium-term cycles, this paper considers house price, gross domestic product per head and the ratio of the two for the UK over 1955–2020. It shown that, although the synchronisation of business and medium-term cycles is associated with the largest price and ratio events, there a shift of volatility from the former to the latter cycle range, which begins in the 1980s. The medium-term cycles are closely aligned. Indeed, the phase-leading role of income over the others is at odds with current stabilisation policy thinking. The trend in income growth is steady but then, around 2002 peters into a stagnant period. The trend in the house price-income ratio is remarkably stable, but in the era of finance liberalisation, that stability is disrupted both in trend and cycle. This appears to be altered by the adjustment to greater financial accessibility. In effect, this trend traces the amount of debt that an agent's income is expected to service when purchasing a dwelling, or the real interest rate. Any return to the "normal" cost of capital could have a severe impact on borrowers.

Biologically composites offer inspiration for the design of next generation structural materials due to their hierarchical microstructure that allows for, low density, and combination of high stiffness and toughness, currently unmatched by engineering technologies. Here, we investigate biomimetic composite materials that feature the laminated helicoidal architecture of fibers characteristic of the arthropod cuticle, specific to the mantis shrimp, but utilize either glass or Kevlar fibers, with epoxy or urethane matrix materials, to examine their role in impact energy dissipation. Drop weight impact test revealed epoxy-matrix composites exhibited the highest peak load on impact and showed larger degrees of external damage such as delamination and fiber breakage as compared to the urethane-based composites. Moreover, the helicoidal panels, showed a significant reduction in dent depth and better residual compressive strength as compared to the quasi-isotropic design. These findings provide useful insight into the variation of reinforcement and matrix materials for the helicoidal architecture and how it affects the delocalization of damage and improves residual strength.

The COVID-19 pandemic and its handling in Sri Lanka has affected vegetable farmers in numerous ways and these impacts will constrain the country's move towards sustainable development. A field level study with vegetable farmers and key informants was carried out using exploratory research to understand, describe and analyze the impact of COVID-19 on the livelihoods of vegetable farmers and its relevance in achieving SDG 1. Data were supplemented by an extensive literature review. The analysis showed that the pandemic's impact on vegetable farmers in Sri Lanka is multidimensional and will increase vulnerability among vegetable farmers, for the long run. Adapting alternative inputs and marketing strategies, provision of immediate financial support, promoting innovative technology and service provision, and implementing intervention strategies tailored to farmer heterogeneity will improve farmer livelihoods and the prosperity of the sector.

A novel dual cure photopolymerizable system was developed by combining two plant-derived acrylic monomers, acrylated epoxidized soybean oil and vanillin dimethacrylate, as well as the thiol monomer pentaerythritol tetrakis (3-mercaptopropionate). Carefully selected resin composition allowed the researchers to overcome earlier stability/premature polymerization problems and to obtain stable (up to six months at 4 °C) and selectively-polymerizable resin. The resin demonstrated rapid photocuring without an induction period and reached a rigidity of 317.66 MPa, which was more than 20 times higher than that of the other vanillin-based polymers. Improved mechanical properties and thermal stability of the resulting cross-linked photopolymer were obtained compared to similar homo- and copolymers: Young’s modulus reached 4753 MPa, the compression modulus reached 1634 MPa, and the temperature of 10% weight loss was 373 °C. The developed photocurable system was successfully applied in stereolithography and characterized with femtosecond pulsed two-beam initiation threshold measurement for the first time. The polymerization threshold of the investigated polymer was determined to be controlled by the sample temperature, making the footprint of the workstations cheaper, faster, and more reliable.

Over the past decade, what it means to be an academic teacher of English-language literature in Swedish institutions of higher education has changed. As a result of recent political reforms, many literature staff have come to assume the role as teacher educators. To better understand the implications of this development, the article maps the academic qualifications and research interests of English staff who teach on teacher education (TEd) literature courses nationally and their attitudes to TEd teaching. The article is based on data gathered via a semi-closed questionnaire and analysed using content and discourse analysis. It shows that a majority of the study participants are PhD holders in English with a specialisation in literature. Although few staff are qualified teachers and/or are engaged in literature teaching and learning scholarship, several have school teaching experience. Respondent attitudes to the teacher educator role vary, as do the conditions for TEd teaching at different institutions. The findings suggest that respondent expertise and self-identification and their previous TEd teaching experiences are consequential for their attitudes, as is the matter of whether the role requires that they address areas, such as school-oriented teaching and learning theories and practices, in which they lack competence. These findings, the article suggests, have bearing on future strategic discussions in English studies.

Many retrofit projects went wrong in the UK principally because of the application of inappropriate retrofit solutions, which resulted in damp issues, with some leaving houses in worse conditions than pre-retrofit. Various online tools were developed to inform homeowners about the benefits of retrofitting. Prior to this study, little was known about users’ evaluation of these tools and the effects of calculator use. Furthermore, no retrofit tool aims to raise the awareness of homeowners about moisture risks in a retrofit project. The originality of this study is to develop and evaluate an online moisture-safe retrofit decision-making tool for homeowners. The adopted methodology consisted of two phases. Phase one aimed to develop the tool. In phase two, semi-structured interviews were conducted to evaluate the tool. The results indicate that the tool has been well received by homeowners. The tool significantly increased participants’ awareness of moisture risks related to a retrofit project. Most participants considered the tool an eye-opener, while few of them found it scary. However, the tool did not result in an increased willingness to invest in energy efficiency measures. The discouragement was related to high investment costs and long payback periods of some retrofit measures. Based on our findings, we formulate a set of design recommendations to improve the proposed tool and help retrofit calculators, in general, overcome challenges.

Burkholderia pseudomallei is a soil-dwelling organism present throughout the tropics. It is the causative agent of melioidosis, a disease that is believed to kill 89,000 people per year. It is naturally resistant to many antibiotics, requiring at least two weeks of intravenous treatment with ceftazidime, imipenem or meropenem followed by 6 months of orally delivered co-trimoxazole. This places a large treatment burden on the predominantly middle-income nations where the majority of disease occurs. We have established a high-throughput assay for compounds that could be used as a co-therapy to potentiate the effect of ceftazidime, using the related non-pathogenic bacterium Burkholderia thailandensis as a surrogate. Optimization of the assay gave a Z' factor of 0.68. We screened a library of 61,250 compounds and identified 29 compounds with a pIC50 (-log10(IC50)) greater than five. Detailed investigation allowed us to down select to six "best in class" compounds, which included the licensed drug chloroxine. Co-treatment of B. thailandensis with ceftazidime and chloroxine reduced culturable cell numbers by two orders of magnitude over 48 hours, compared to treatment with ceftazidime alone. Hit expansion around chloroxine was performed using commercially available compounds. Minor modifications to the structure abolished activity, suggesting that chloroxine likely acts against a specific target. Finally, an initial study demonstrates the utility of chloroxine to act as a co-therapy to potentiate the effect of ceftazidime against B. pseudomallei. This approach successfully identified potential co-therapies for a recalcitrant Gram-negative bacterial species. Our assay could be used more widely to aid in chemotherapy to treat infections caused by these bacteria.

Of the 16 non-structural proteins (Nsps) encoded by SARS CoV-2, Nsp3 is the largest and plays important roles in the viral life cycle. Being a large, multidomain, transmembrane protein, Nsp3 has been the most challenging Nsp to characterize. Encoded within Nsp3 is the papain-like protease domain (PLpro) that cleaves not only the viral polypeptide but also K48-linked polyubiquitin and the ubiquitin-like modifier, ISG15, from host cell proteins. We here compare the interactors of PLpro and Nsp3 and find a largely overlapping interactome. Intriguingly, we find that near full length Nsp3 is a more active protease compared to the minimal catalytic domain of PLpro. Using a MALDI-TOF based assay, we screen 1971 approved clinical compounds and identify five compounds that inhibit PLpro with IC50s in the low micromolar range but showed cross reactivity with other human deubiquitinases and had no significant antiviral activity in cellular SARS-CoV-2 infection assays. We therefore looked for alternative methods to block PLpro activity and engineered competitive nanobodies that bind to PLpro at the substrate binding site with nanomolar affinity thus inhibiting the enzyme. Our work highlights the importance of studying Nsp3 and provides tools and valuable insights to investigate Nsp3 biology during the viral infection cycle.

The UK has one of the least energy-efficient housing stocks in Europe. By 2030, the emissions from UK homes need to fall by at least 24% from 1990 levels to meet the UK’s ambitious goal, which is reaching net-zero emissions. The originality of this paper is to apply the building typology approach to predict energy savings of the UK housing stock under a step-by-step energy retrofit scenario, targeting the Passive House Standard for refurbishments of existing buildings, namely the EnerPHit “Quality-Approved Energy Retrofit with Passive House Components.” The typologies consist of twenty reference buildings, representative of five construction ages and four building sizes. The energy balance of the UK residential buildings was created and validated against statistical data. A building stock retrofit plan specifying the order in which to apply energy efficiency measures was elaborated, and energy savings were calculated. The predicted total energy demand for the UK residential building stock for the year 2022 is 37.7 MTOE, and the carbon emissions estimation is 65.33 MtCO2e. The energy-saving potential is 87%, and carbon reductions are about 76%, considering all the steps of renovation applied. It has been demonstrated that the step that provides the biggest savings across the housing stock is the one that involves replacing windows, draught-proofing, and installing mechanical ventilation with heat recovery.

Background: PF-06649751 is a novel, oral, non-catechol-based, D1/D5 dopamine receptor partial agonist under investigation for the treatment of motor symptoms associated with Parkinson’s disease. Methods: A 15-week, phase II, double-blind, placebo-controlled clinical trial was conducted to assess the efficacy and safety of flexible-dose PF-06649751 in subjects with early stage Parkinson’s disease (ClinicalTrials.gov identifier: NCT02847650). Results: Enrollment was terminated early for reasons unrelated to the trial. Overall, 57 subjects received study medication (PF-06649751 = 29; placebo = 28) and 47 completed the study (PF-06649751 = 25; placebo = 22). Despite early termination, the study met its primary endpoint with the PF-06649751 group showing statistically significant improvement from baseline in the Movement Disorder Society-Unified Parkinson’s Disease Rating Scale (MDS-UPDRS) Part III score at week 15 compared with placebo. Mean (SE) change in MDS-UPDRS Part III score was −9.0 (1.54) for PF-06649751 and −4.3 (1.65) for placebo. This corresponds to an improvement versus placebo of 4.8 for the PF-06649751 group (two-sided p = 0.0407; 90% CI = 1.0, 8.6). Statistically significant improvement in MDS-UPDRS-III score was also observed at all assessment time points prior to week 15. The safety profile of PF-06649751 was similar to that observed in prior studies, with the majority of adverse events (AEs) reported as mild or moderate. The most common AEs in the PF-06649751 group were nausea, headache, dry mouth, somnolence, and tremor. Conclusions: Once-daily dosing of oral PF-06649751 resulted in significant improvement of motor symptoms and was generally well tolerated in subjects with early stage Parkinson’s disease.

B cells are critical for promoting autoimmunity and the success of B cell depletion therapy in rheumatoid arthritis (RA) confirms their importance in driving chronic inflammation. Whilst disease specific autoantibodies are useful diagnostically, our understanding of the pathogenic B cell repertoire remains unclear. Defining it would lead to novel insights and curative treatments. To address this, we have undertaken the largest study to date of over 150 RA patients, utilizing next generation sequencing (NGS) to analyze up to 200,000 BCR sequences per patient. The full-length antigen-binding variable region of the heavy chain (IgGHV) of the IgG B cell receptor (BCR) were sequenced. Surprisingly, RA patients do not express particular clonal expansions of B cells at diagnosis. Rather they express a polyclonal IgG repertoire with a significant increase in BCRs that have barely mutated away from the germline sequence. This pattern remains even after commencing disease modifying therapy. These hypomutated BCRs are expressed by TNF-alpha secreting IgG+veCD27−ve B cells, that are expanded in RA peripheral blood and enriched in the rheumatoid synovium. A similar B cell repertoire is expressed by patients with Sjögren's syndrome. A rate limiting step in the initiation of autoimmunity is the activation of B cells and this data reveals that a sizeable component of the human autoimmune B cell repertoire consists of polyclonal, hypomutated IgG+ve B cells, that may play a critical role in driving chronic inflammation.

The 2.02 ha site containing the Category B listed Walled Garden at Benmore is currently the subject of a major redesign proposal and active fundraising programme. The purpose of this article is to raise the profile of the project by investigating and highlighting the historical development of the site. This retrospective study is also intended as a support to contemporary redevelopment plans and as a demonstration of how the past underpins and informs the future. I am frankly and absolutely for a formal garden … It is a small piece of ground enclosed by walls … There is not the least attempt to imitate natural scenery (Phillpotts, 1906, p. 54).

Landscape modification associated with agricultural intensification has brought considerable challenges for the sustainable development of New Zealand hill country farms. Addressing these challenges requires an appropriate approach to support farmers and design a better landscape that can have beneficial environmental outcomes whilst ensuring continued profitability. In this paper we suggest using geodesign and theories drawn from landscape ecology to plan and design multifunctional landscapes that offer improved sustainability for hill country farm systems and landscapes in New Zealand. This approach suggests that better decisions can be made by considering the major landscape services that are, and could be, provided by the landscapes in which these farm systems are situated. These important services should be included in future landscape design of hill country by creating a patterning and configuration of landscape features that actively maintains or restores important landscape functioning. This will help to improve landscape health and promote landscape resilience in the face of climate change. Through illustrating the potential of this type of approach for wider adoption we believe that the proposed conceptual framework offers a valuable reference for sustainable farm system design that can make an important contribution to advancing environmental management globally as well as in New Zealand.

The chronic autoimmune inflammatory diseases, systemic lupus erythematosus and Sjogren’s syndrome, develop when tolerance to apoptotic cells (ACs) is lost. We have previously reported that this tolerance is maintained by innate-like, IL-10 secreting regulatory B cells. Two questions remained. First, do these regulatory B cells belong predominantly to a single subset of steady-state B cells and second, what is their specificity? We report here that innate-like B cells with markers characteristic for B1a cells (CD43+veCD19hiCD5+veIgMhiIgDlo) constitute 80% of splenic and 96% of peritoneal B cells that respond to ACs by secreting IL-10. AC responsive B1a cells secrete self-reactive natural antibodies (NAbs) and IL-10, which is augmented by toll-like receptor (TLR) 7 or TLR9 stimulation. In so doing, they both accelerate the clearance of dying cells by macrophages and inhibit their potential to mount proinflammatory immune responses. While B1a cells make prolonged contact with ACs, they do not require TIM1 or complement to mediate their regulatory function. In an animal model of neural inflammation (experimental autoimmune encephalomyelitis), just 105 activated B1a B cells was sufficient to restrain inflammation. Activated B1a B cells also induced antigen-specific T cells to secrete IL-10. Hence, regulatory B1a cells specifically recognize and augment tolerance to apoptotic self via IL-10 and NAbs; but once activated, can also prevent autoimmune mediated inflammation.

Deer damage to plants within gardens is a common and widespread problem. Gardens adjacent to habitats with natural deer populations are particularly at risk. This paper provides a brief history and description of various types of deer damage frequently encountered at Benmore Botanic Garden on the west coast of Scotland. Thereafter follows a summary of a localised plant protection system, which has been developed through much experimentation over the last few years, to reduce deer damage to woody plants throughout the Garden.

T follicular helper cells (Tfh) provide crucial signals for germinal center (GC) formation, but Tfh populations are heterogeneous. While PD1hi Tfh are important in the GC response, the function of the PD1lo Tfh-like subset is unknown. We show that these cells, like the PD1hi GC–Tfh, depend upon B cells; however, their entry to follicles is independent of CXCR5 or cognate interactions with B cells. The differentiation into PD1hi Tfh is dependent on MHC class II interactions with B cells and requires CXCR5. Our data suggest a Tfh differentiation pathway that is initially B cell-independent, then dependent on non-cognate B cell interactions, and finally following cognate interaction with B cells and CXCR5-ligands allows the formation of GC–Tfh. The PD1lo Tfh-like cells make early cytokine responses and may represent precursors of CD4 memory cells.

A supply of pure, intact oil bodies is essential for carrying out morphological and biochemical studies of these plant organelles and exploring their application. Preparation requires a carefully controlled breakage of plant cells, followed by separation of the oil bodies from cytoplasm and cell debris. This paper focuses on the recovery and characterisation of oil bodies from pecan nuts where no work has been published to date. The results showed that soaking softens the nut tissue and appears to reduce the damage to oil bodies during grinding and centrifugal force must be carefully selected to minimise oil bodies damage on recovery. A 24 h soaking time coupled with a 5500 RCF recovery force allows for the recovery of intact pecan nut oil bodies.

Background. The myeloid enzyme 12/15-lipoxygenase (LOX), which generates bioactive oxidized lipids, has been implicated in numerous inflammatory diseases, with several studies demonstrating an improvement in pathology in mice lacking the enzyme. However, the ability of 12/15-LOX to directly regulate B cell function has not been studied. Methods. The influence of 12/15-LOX on B cell phenotype and function, and IgM generation, was compared using wildtype (WT) and 12/15-LOX (Alox15-/-) deficient mice. The proliferative and functional capacity of splenic CD19+ B cells was measured in vitro in response to various toll-like receptor agonists. Results. WT and Alox15-/- displayed comparable responses. However in vivo, splenic B cell numbers were significantly elevated in Alox15-/- mice with a corresponding elevation in titres of total IgM in lung, gut and serum, and lower serum IgM directed against the 12/15-LOX product, 12-hydroxyeicosatetraenoic acid-phosphatidylethanolamine (HETE-PE). Discussion. Myeloid 12/15-LOX can regulate B cell numbers and innate immune antibody levels in vivo, potentially contributing to its ability to regulate inflammatory disease. Furthermore, the alterations seen in 12/15-LOX deficiency likely result from changes in the equilibrium of the immune system that develop from birth. Further studies in disease models are warranted to elucidate the contribution of 12/15-LOX mediated alterations in B cell numbers and innate immune antibody generation to driving inflammation in vivo.

Leishmaniasis is a neglected disease that is caused by different species of the protozoan parasite Leishmania, and it currently affects 12 million people worldwide. The antileishmanial therapeutic arsenal remains very limited in number and efficacy, and there is no vaccine for this parasitic disease. One pathway that has been genetically validated as an antileishmanial drug target is the biosynthesis of uridine diphosphate-glucose (UDP-Glc), and its direct derivative UDP-galactose (UDP-Gal). De novo biosynthesis of these two nucleotide sugars is controlled by the specific UDP-glucose pyrophosphorylase (UGP). Leishmania parasites additionally express a UDP-sugar pyrophosphorylase (USP) responsible for monosaccharides salvage that is able to generate both UDP-Gal and UDP-Glc. The inactivation of the two parasite pyrophosphorylases UGP and USP, results in parasite death. The present study reports on the identification of structurally diverse scaffolds for the development of USP inhibitors by fragment library screening. Based on this screening, we selected a small set of commercially available compounds, and identified molecules that inhibit both Leishmania major USP and UGP, with a half-maximal inhibitory concentration in the 100 µM range. The inhibitors were predicted to bind at allosteric regulation sites, which were validated by mutagenesis studies. This study sets the stage for the development of potent USP inhibitors.