Identification, prevalence and impacts of viral diseases of UK winter wheat

The potential for viruses to be causing the plateau in the yield of UK wheat (Triticum aestivum) was investigated. Mechanical inoculation of Cynosurus mottle virus to wheat cv. Scout and cv. Gladiator caused 83% and 58% reduction in the number of grains produced, highlighting the potential of viruses to cause disease and yield loss. Viruses historically detected in cereals in the UK were not found to be prevalent following real time reverse transcriptase polymerase chain reaction testing of 1,356 UK wheat samples from 2009-2012 using eleven assasys developed in the project. This included an assay for Cynosurus mottle virus, which was based on its complete genome sequence which was obtained for the first time in this project. Viruses detected were Barley yellow dwarf virus-MAV (6 samples) (BYDV-MAV), Barley yellow dwarf virus-PAV (6 samples) (BYDV-PAV) and Soil-borne cereal mosaic virus (12 samples) (SBCMV). There was a higher prevalence of viruses in the south, thought to be due to warmer temperatures which benefitted insect vectors and the molecular processes of infection. Viruses were most commonly detected in the variety JB Diego, perhaps because this variety has no known resistance to viruses. The low prevalence of known viruses could also have been because they were outcompeted or replaced by previously unknown ones. Next generation sequencing was used to test 120 samples from an organic site, including wheat, weeds and insects, to search for novel viruses. Testing of twelve storage regimes for insect traps using BYDV-PAV infected Sitobion avenae for recovery of PCR amplifiable RNA using 18S rRNA and BYDV-PAV assays found that 0.5 M EDTA was the most successful regime which was therefore used in the collection of samples for sequencing. Known viruses such as BYDV-PAV were detected along with some additional potentially novel viruses (eight possibly novel viruses or strains of viruses with four in wheat). One such virus was apparently present in 25% of all wheat samples tested, making it potentially very significant. This could be important for unlocking the yield potential of wheat because it could be a cryptic virus which is highly prevalent. In order to control the spread of viruses their methods of transmission must be understood, therefore testing of seeds and resulting plants from Cynosurus mottle virus infected material was done. Tests did not detect the virus, therefore it was concluded that seed transmission does not occur. However, further tests are required. In conclusion this study indicates that known viruses are not currently a major problem for UK winter wheat. However, novel viruses that are a problem may be detected in the future perhaps by next generation sequencing. Additonal viruses from abroad would add to the threat. The impact of all viruses in wheat may be greater in the future due to climate change

Identification, prevalence and impacts of viral diseases of UK winter wheat

The potential for viruses to be causing the plateau in the yield of UK wheat (Triticum aestivum) was investigated. Mechanical inoculation of Cynosurus mottle virus to wheat cv. Scout and cv. Gladiator caused 83% and 58% reduction in the number of grains produced, highlighting the potential of viruses to cause disease and yield loss. Viruses historically detected in cereals in the UK were not found to be prevalent following real time reverse transcriptase polymerase chain reaction testing of 1,356 UK wheat samples from 2009-2012 using eleven assasys developed in the project. This included an assay for Cynosurus mottle virus, which was based on its complete genome sequence which was obtained for the first time in this project. Viruses detected were Barley yellow dwarf virus-MAV (6 samples) (BYDV-MAV), Barley yellow dwarf virus-PAV (6 samples) (BYDV-PAV) and Soil-borne cereal mosaic virus (12 samples) (SBCMV). There was a higher prevalence of viruses in the south, thought to be due to warmer temperatures which benefitted insect vectors and the molecular processes of infection. Viruses were most commonly detected in the variety JB Diego, perhaps because this variety has no known resistance to viruses. The low prevalence of known viruses could also have been because they were outcompeted or replaced by previously unknown ones. Next generation sequencing was used to test 120 samples from an organic site, including wheat, weeds and insects, to search for novel viruses. Testing of twelve storage regimes for insect traps using BYDV-PAV infected Sitobion avenae for recovery of PCR amplifiable RNA using 18S rRNA and BYDV-PAV assays found that 0.5 M EDTA was the most successful regime which was therefore used in the collection of samples for sequencing. Known viruses such as BYDV-PAV were detected along with some additional potentially novel viruses (eight possibly novel viruses or strains of viruses with four in wheat). One such virus was apparently present in 25% of all wheat samples tested, making it potentially very significant. This could be important for unlocking the yield potential of wheat because it could be a cryptic virus which is highly prevalent. In order to control the spread of viruses their methods of transmission must be understood, therefore testing of seeds and resulting plants from Cynosurus mottle virus infected material was done. Tests did not detect the virus, therefore it was concluded that seed transmission does not occur. However, further tests are required. In conclusion this study indicates that known viruses are not currently a major problem for UK winter wheat. However, novel viruses that are a problem may be detected in the future perhaps by next generation sequencing. Additonal viruses from abroad would add to the threat. The impact of all viruses in wheat may be greater in the future due to climate change

Microglia in Close Vicinity of Glioma Cells: Correlation Between Phenotype and Metabolic Alterations

Microglia are immune cells within the central nervous system. In brain-developing tumors, gliomas are able to silence the defense and immune functions of microglia, a phenomenon which strongly contributes to tumor progression and treatment resistance. Being activated and highly motile, microglia infiltrate tumors and secrete macrophagic chemoattractant factors. Thereafter, the tumor cells shut down their immune properties and stimulate the microglia to release tumor growth-promoting factors. The result of such modulation is that a kind of symbiosis occurs between microglia and tumor cells, in favor of tumor growth. However, little is known about microglial phenotype and metabolic modifications in a tumoral environment. Co-cultures were performed using CHME5 microglia cells grown on collagen beads or on coverslips and placed on monolayer of C6 cells, limiting cell/cell contacts. Phagocytic behavior and expression of macrophagic and cytoskeleton markers were monitored. Respiratory properties and energetic metabolism were also studied with regard to the activated phenotype of microglia. In co-cultures, transitory modifications of microglial morphology and metabolism were observed linked to a concomitant transitory increase of phagocytic properties. Therefore, after 1 h of co-culture, microglia were activated but when longer in contact with tumor cells, phagocytic properties appear silenced. Like the behavior of the phenotype, microglial respiration showed a transitory readjustment although the mitochondria maintained their perinuclear relocation. Nevertheless, the energetic metabolism of the microglia was altered, suggesting a new energetic steady state. The results clearly indicate that like the depressed immune properties, the macrophagic and metabolic status of the microglia is quickly driven by the glioma environment, despite short initial phagocytic activation. Such findings question the possible contribution of diffusible tumor factors to the microglial metabolism

Interlaboratory evaluation of rat hepatic gene expression changes induced by methapyrilene.

Several studies using microarrays have shown that changes in gene expression provide information about the mechanism of toxicity induced by xenobiotic agents. Nevertheless, the issue of whether gene expression profiles are reproducible across different laboratories remains to be determined. To address this question, several members of the Hepatotoxicity Working Group of the International Life Sciences Institute Health and Environmental Sciences Institute evaluated the liver gene expression profiles of rats treated with methapyrilene (MP). Animals were treated at one facility, and RNA was distributed to five different sites for gene expression analysis. A preliminary evaluation of the number of modulated genes uncovered striking differences between the five different sites. However, additional data analysis demonstrated that these differences had an effect on the absolute gene expression results but not on the outcome of the study. For all users, unsupervised algorithms showed that gene expression allows the distinction of the high dose of MP from controls and low dose. In addition, the use of a supervised analysis method (support vector machines) made it possible to correctly classify samples. In conclusion, the results show that, despite some variability, robust gene expression changes were consistent between sites. In addition, key expression changes related to the mechanism of MP-induced hepatotoxicity were identified. These results provide critical information regarding the consistency of microarray results across different laboratories and shed light on the strengths and limitations of expression profiling in drug safety analysis

Affinity maturation generates greatly improved xyloglucan-specific carbohydrate binding modules

<p>Abstract</p> <p>Background</p> <p>Molecular evolution of carbohydrate binding modules (CBM) is a new approach for the generation of glycan-specific molecular probes. To date, the possibility of performing affinity maturation on CBM has not been investigated. In this study we show that binding characteristics such as affinity can be improved for CBM generated from the CBM4-2 scaffold by using random mutagenesis in combination with phage display technology.</p> <p>Results</p> <p>Two modified proteins with greatly improved affinity for xyloglucan, a key polysaccharide abundant in the plant kingdom crucial for providing plant support, were generated. Both improved modules differ from other existing xyloglucan probes by binding to galactose-decorated subunits of xyloglucan. The usefulness of the evolved binders was verified by staining of plant sections, where they performed better than the xyloglucan-binding module from which they had been derived. They discriminated non-fucosylated from fucosylated xyloglucan as shown by their ability to stain only the endosperm, rich in non-fucosylated xyloglucan, but not the integument rich in fucosylated xyloglucan, on tamarind seed sections.</p> <p>Conclusion</p> <p>We conclude that affinity maturation of CBM selected from molecular libraries based on the CBM4-2 scaffold is possible and has the potential to generate new analytical tools for detection of plant carbohydrates.</p

Harnessing the PRECISE network as a platform to strengthen global capacity for maternal and child health research in sub-Saharan Africa

It is widely acknowledged across the global health sector that research programmes need to be designed and implemented in a way that maximise opportunities for strengthening local capacity. This paper examines how the United Kingdom Research and Innovation (UKRI) Grand Challenges Research Fund (GCRF) funded PRECISE (PREgnancy Care Integrating translational Science, Everywhere) Network has been established as a platform to strengthen global capacity for research focused on the improvement of maternal, fetal and newborn health in subSaharan Africa. Best practice principles outlined in an ESSENCE on Health Research report have been considered in relation to the PRECISE Network capacity-building activities described in this paper. These activities are described at the individual, programmatic and institutional levels, and successes, challenges and recommendations for future work are outlined. The paper concludes that the PRECISE leadership have an opportunity to review and refresh activity plans for capacity building at this stage in the project to build on achievements to date

Role of MALDI-MSI in combination with 3D tissue models for early stage efficacy and safety testing of drugs and toxicants

Introduction: Three-dimensional (3D) cell cultures have become increasingly important materials to investigate biological processes and drug efficacy and toxicity. The ability of 3D cultures to mimic the physiology of primary tissues and organs in the human body enables further insight into cellular behavior and is hence highly desirable in early-stage drug development. Analyzing the spatial distribution of drug compounds and endogenous molecules provides an insight into the efficacy of a drug whilst simultaneously giving information on biological responses. Areas Covered: In this review we will examine the main 3D cell culture systems employed and applications, which describe their integration with mass spectrometry imaging (MSI). Expert Opinion: MSI is a powerful technique that can map a vast range of molecules simultaneously in tissues without the addition of labels that can provide insights into the efficacy and safety of a new drug. The combination of MSI and 3D cell cultures has emerged as a promising tool in early-stage drug analysis. However, the most common administration route for pharmaceutical drugs is via oral delivery. The use of MSI in combination with models of the GI tract is an area that has been little explored to date, the reasons for this are discussed

Emerging applications in mass spectrometry imaging; enablers and roadblocks

© 2020 The Authors. Mass spectrometry imaging (MSI) is a powerful and versatile technique able to investigate the spatial distribution of multiple non-labelled endogenous and exogenous analytes simultaneously, within a wide range of samples. Over the last two decades, MSI has found widespread application for an extensive range of disciplines including pre-clinical drug discovery, clinical applications and human identification for forensic purposes. Technical advances in both instrumentation and software capabilities have led to a continual increase in the interest in MSI; however, there are still some limitations. In this review, we discuss the emerging applications in MSI that significantly impact three key areas of mass spectrometry (MS) research—clinical, pre-clinical and forensics—and roadblocks to the expansion of use of MSI in these areas

Mutation of the Diamond-Blackfan Anemia Gene Rps7 in Mouse Results in Morphological and Neuroanatomical Phenotypes

The ribosome is an evolutionarily conserved organelle essential for cellular function. Ribosome construction requires assembly of approximately 80 different ribosomal proteins (RPs) and four different species of rRNA. As RPs co-assemble into one multi-subunit complex, mutation of the genes that encode RPs might be expected to give rise to phenocopies, in which the same phenotype is associated with loss-of-function of each individual gene. However, a more complex picture is emerging in which, in addition to a group of shared phenotypes, diverse RP gene-specific phenotypes are observed. Here we report the first two mouse mutations (Rps7(Mtu) and Rps7(Zma)) of ribosomal protein S7 (Rps7), a gene that has been implicated in Diamond-Blackfan anemia. Rps7 disruption results in decreased body size, abnormal skeletal morphology, mid-ventral white spotting, and eye malformations. These phenotypes are reported in other murine RP mutants and, as demonstrated for some other RP mutations, are ameliorated by Trp53 deficiency. Interestingly, Rps7 mutants have additional overt malformations of the developing central nervous system and deficits in working memory, phenotypes that are not reported in murine or human RP gene mutants. Conversely, Rps7 mouse mutants show no anemia or hyperpigmentation, phenotypes associated with mutation of human RPS7 and other murine RPs, respectively. We provide two novel RP mouse models and expand the repertoire of potential phenotypes that should be examined in RP mutants to further explore the concept of RP gene-specific phenotypes.This research was supported in part by the Intramural Research Program of NHGRI, NIH, and the Wellcome Trust and by NHMRC Australia grant 366746. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript