An NADPH-Dependent Genetic Switch Regulates Plant Infection by the Rice Blast Fungus

To cause rice blast disease, the fungus Magnaporthe oryzae breaches the tough outer cuticle of the rice leaf by using specialized infection structures called appressoria. These cells allow the fungus to invade the host plant and proliferate rapidly within leaf tissue. Here, we show that a unique NADPH-dependent genetic switch regulates plant infection in response to the changing nutritional and redox conditions encountered by the pathogen. The biosynthetic enzyme trehalose-6-phosphate synthase (Tps1) integrates control of glucose-6-phosphate metabolism and nitrogen source utilization by regulating the oxidative pentose phosphate pathway, the generation of NADPH, and the activity of nitrate reductase. We report that Tps1 directly binds to NADPH and, thereby, regulates a set of related transcriptional corepressors, comprising three proteins, Nmr1, Nmr2, and Nmr3, which can each bind NADP. Targeted deletion of any of the Nmr-encoding genes partially suppresses the nonpathogenic phenotype of a Δtps1 mutant. Tps1-dependentNmr corepressors control the expression of a set of virulence-associated genes that are derepressed during appressorium-mediated plant infection. When considered together, these results suggest that initiation of rice blast disease by M. oryzae requires a regulatory mechanism involving an NADPH sensor protein, Tps1, a set of NADP-dependent transcriptional corepressors, and the nonconsuming interconversion ofNADPHandNADPacting as signal transducer

Reflections and Experiences of a Co-Researcher involved in a Renal Research Study

Background Patient and Public Involvement (PPI) is seen as a prerequisite for health research. However, current Patient and public involvement literature has noted a paucity of recording of patient and public involvement within research studies. There have been calls for more recordings and reflections, specifically on impact. Renal medicine has also had similar criticisms and any reflections on patient and public involvement has usually been from the viewpoint of the researcher. Roles of patient and public involvement can vary greatly from sitting on an Advisory Group to analysing data. Different PPI roles have been described within studies; one being a co-researcher. However, the role of the co-researcher is largely undefined and appears to vary from study to study. Methods The aims of this paper are to share one first time co-researcher's reflections on the impact of PPI within a mixed methods (non-clinical trial) renal research study. A retrospective, reflective approach was taken using data available to the co-researcher as part of the day-to-day research activity. Electronic correspondence and documents such as meeting notes, minutes, interview thematic analysis and comments on documents were re-examined. The co-researcher led on writing this paper. Results This paper offers a broad definition of the role of the co-researcher. The co-researcher reflects on undertaking and leading on the thematic analysis of interview transcripts, something she had not previously done before. The co-researcher identified a number of key themes; the differences in time and responsibility between being a coresearcher and an Advisory Group member; how the role evolved and involvement activities could match the co-researchers strengths (and the need for flexibility); the need for training and support and lastly, the time commitment. It was also noted that it is preferable that a co-researcher needs to be involved from the very beginning of the grant application. Conclusions The reflections, voices and views of those undertaking PPI has been largely underrepresented in the literature. The role of co-researcher was seen to be rewarding but demanding, requiring a large time commitment. It is hoped that the learning from sharing this experience will encourage others to undertake this role, and encourage researchers to reflect on the needs of those involved.Peer reviewedFinal Published versio

The place of strategic environmental assessment in the privatised electricity industry

The private sector has given relatively little attention to the emergence of strategic environmental assessment (SEA); even recently privatised utilities, where SEA might be deemed particularly appropriate, and whose activities are likely to fall within the scope of the European Union SEA Directive, have shown less interest than might be expected. However, the global trend towards the privatisation of state-owned enterprises makes the adaptation of SEA towards these industries all the more pressing. This paper addresses the place that SEA might take within the electricity sector, taking the privatised UK electricity industry as an example. Particular challenges are posed by the radical restructuring of the industry, designed to introduce competitive behaviour, making the development of comprehensive SEA processes problematic, and requiring SEA to be placed in the context of corporate environmental policy and objectives.</p

Coccolithophores: Functional Biodiversity, Enzymes and Bioprospecting

Emiliania huxleyi is a single celled, marine phytoplankton with global distribution. As a key species for global biogeochemical cycling, a variety of strains have been amassed in various culture collections. Using a library consisting of 52 strains of E. huxleyi and an ‘in house’ enzyme screening program, we have assessed the functional biodiversity within this species of fundamental importance to global biogeochemical cycling, whilst at the same time determining their potential for exploitation in biocatalytic applications. Here, we describe the screening of E. huxleyi strains, as well as a coccolithovirus infected strain, for commercially relevant biocatalytic enzymes such as acid/alkali phosphodiesterase, acid/alkali phosphomonoesterase, EC1.1.1-type dehydrogenase, EC1.3.1-type dehydrogenase and carboxylesterase

A high-sensitivity electrochemiluminescence-based ELISA for the measurement of the oxidative stress biomarker, 3-nitrotyrosine, in human blood serum and cells

The generation of 3-nitrotyrosine, within proteins, is a post-translational modification resulting from oxidative or nitrative stress. It has been suggested that this modification could be used as a biomarker for inflammatory diseases. Despite the superiority of mass spectrometry-based determinations of nitrotyrosine, in a high-throughput clinical setting the measurement of nitrotyrosine by an enzyme-linked immunosorbent assay (ELISA) is likely to be more cost-effective. ELISAs offer an alternative means to detect nitrotyrosine, but many commercially available ELISAs are insufficiently sensitive to detect nitrotyrosine in healthy human serum. Here, we report the development, validation and clinical application of a novel electrochemiluminescence-based ELISA for nitrotyrosine which provides superior sensitivity (e.g. a 50-fold increase in sensitivity compared with one of the tested commercial colorimetric ELISAs). This nitrotyrosine ELISA has the following characteristics: a lower limit of quantitation of 0.04 nM nitrated albumin equivalents; intra- and inter-assay coefficients of variation of 6.5% and 11.3%, respectively; a mean recovery of 106 ± 3% and a mean linearity of 0.998 ± 0.001. Far higher nitration levels were measured in normal human blood cell populations when compared to plasma. Mass spectrometry was used to validate the new ELISA method. The analysis of the same set of chemically modified albumin samples using the ELISA method and mass spectrometry showed good agreement for the relative levels of nitration present in each sample. The assay was applied to serum samples from patients undergoing elective surgery which induces the human inflammatory response. Matched samples were collected before and one day after surgery. An increase in nitration was detected following surgery (median (IQR): 0.59 (0.00–1.34) and 0.97 (0.00–1.70) nitrotyrosine (fmol of nitrated albumin equivalents/mg protein) for pre- and post-surgery respectively. The reported assay is suitable for nitrotyrosine determination in patient serum samples, and may also be applicable as a means to determine oxidative stress in primary and cultured cell populations

Reconceptualizing conservation

Early definitions of conservation focused largely on the end goals of protection or restoration of nature, and the various disciplinary domains that contribute to these ends. Conservation science and practice has evolved beyond being focused on just issues of scarcity and biodiversity decline. To better recognize the inherent links between human behaviour and conservation, “success” in conservation is now being defined in terms that include human rights and needs. We also know that who engages in conservation, and how, dictates the likelihood that conservation science will be embraced and applied to yield conservation gains. Here we present ideas for reconceptualizing conservation. We emphasize the HOW in an attempt to reorient and repurpose the term in ways that better reflect what contemporary conservation is or might aspire to be. To do so, we developed an acrostic using the letters in the term “CONSERVATION” with each serving as an adjective where C = co-produced, O = open, N = nimble, S = solutions-oriented, E = empowering, R = relational, V = values-based, A = actionable, T = transdisciplinary, I = inclusive, O = optimistic, and N = nurturing. For each adjective, we briefly describe our reasoning for its selection and describe how it contributes to our vision of conservation. By reconceptualizing conservation we have the potential to center how we do conservation in ways that are more likely to result in outcomes that benefit biodiversity while also being just, equitable, inclusive, and respectful of diverse rights holders, knowledge holders, and other actors. We hope that this acrostic will be widely adopted in training to help the next generation of conservation researchers and practitioners keep in mind what it will take to make their contributions effective and salient

The ‘appropriate adult’: what they do and what they should do in police interviews with mentally disordered suspects

Background: In almost all countries worldwide, the first point of contact with the criminal justice system is with the police. A large proportion of these individuals may have vulnerabilities, such as mental health difficulties. Given the complexities associated with vulnerable suspects, such interviews may be compromised, which could lead to a miscarriage of justice. In England and Wales, the Police and Criminal Evidence Act (PACE) 1984 and its accompanying Codes of Practice lay down requirements for interviewing vulnerable suspects and provide for attendance of ‘appropriate adults’ to support communication between police and the vulnerable suspect. To date, however, their role has been under-researched. Aims/Hypotheses: To explore the role of appropriate adults in police interviews and test the hypotheses first that appropriate adults more commonly remain passive during interview than expected from guidance and, secondly, that any interventions are more likely than not to follow examples in current guidance. Methods: Transcripts of police interviews conducted with suspects with possible mental disorder and an appropriate adult present (N = 27) were analysed using a specially developed coding framework. Results: Appropriate adults were significantly more likely to remain passive than to intervene, even when current guidance would suggest intervention. When they did intervene, however, such interventions were significantly more likely than not to follow from guidance and the vulnerable suspect’s needs. Conclusions/Implications for practice: In our sample, appropriate adults were not fulfilling their role as outlined in the Police and Criminal Evidence Act (PACE) 1984 and accompanying Codes of Practice, specifically, they appeared to know what to do but not when to do it. There is a heightened risk of a miscarriage of justice in such circumstances without improvements