Role of the type VI secretion systems during disease interactions of Erwinia amylovora with its plant host

Background: Type VI secretion systems (T6SS) are widespread among Gram-negative bacteria and have a potential role as essential virulence factors or to maintain symbiotic interactions. Three T6SS gene clusters were identified in the genome of E. amylovora CFBP 1430, of which T6SS-1 and T6SS-3 represent complete T6SS machineries, while T6SS-2 is reduced in its gene content. Results: To assess the contribution of T6SSs to virulence and potential transcriptomic changes of E. amylovora CFBP 1430, single and double mutants in two structural genes were generated for T6SS-1 and T6SS-3. Plant assays showed that mutants in T6SS-3 were slightly more virulent in apple shoots while inducing less disease symptoms on apple flowers, indicating that T6SSs have only a minor effect on virulence of E. amylovora CFBP 1430. The mutations led under in vitro conditions to the differential expression of type III secretion systems, iron acquisition, chemotaxis, flagellar, and fimbrial genes. Comparison of the in planta and in vitro transcriptome data sets revealed a common differential expression of three processes and a set of chemotaxis and motility genes. Additional experiments proved that T6SS mutants are impaired in their motility. Conclusion: These results suggest that the deletion of T6SSs alters metabolic and motility processes. Nevertheless, the difference in lesion development in apple shoots and flower necrosis of T6SS mutants was indicative that T6SSs influences the disease progression and the establishment of the pathogen on host plants

Transcriptional profile of Salmonella enterica subsp. enterica serovar Weltevreden during alfalfa sprout colonization

Sprouted seeds represent a great risk for infection by human enteric pathogens because of favourable growth conditions for pathogens during their germination. The aim of this study was to identify mechanisms of interactions of Salmonella enterica subsp. enterica Weltevreden with alfalfa sprouts. RNA-seq analysis of S. Weltevreden grown with sprouts in comparison with M9-glucose medium showed that among a total of 4158 annotated coding sequences, 177 genes (4.3%) and 345 genes (8.3%) were transcribed at higher levels with sprouts and in minimal medium respectively. Genes that were higher transcribed with sprouts are coding for proteins involved in mechanisms known to be important for attachment, motility and biofilm formation. Besides gene expression required for phenotypic adaption, genes involved in sulphate acquisition were higher transcribed, suggesting that the surface on alfalfa sprouts may be poor in sulphate. Genes encoding structural and effector proteins of Salmonella pathogenicity island 2, involved in survival within macrophages during infection of animal tissue, were higher transcribed with sprouts possibly as a response to environmental conditions. This study provides insight on additional mechanisms that may be important for pathogen interactions with sprouts

The construct validity of the Dutch personality inventory for DSM-5 personality disorders (PID-5) in a clinical sample

The factor structure and the convergent validity of the Personality Inventory for DSM-5 (PID-5), a self-report questionnaire designed to measure personality pathology as advocated in the fifth edition, Section III of Diagnostic and Statistical Manual of Mental Disorders (DSM-5), are already demonstrated in general population samples, but need replication in clinical samples. In 240 Flemish inpatients, we examined the factor structure of the PID-5 by means of exploratory structural equation modeling. Additionally, we investigated differences in PID-5 higher order domain scores according to gender, age and educational level, and explored convergent and discriminant validity by relating the PID-5 with the Dimensional Assessment of Personality PathologyBasic Questionnaire and by comparing PID-5 scores of inpatients with and without a DSM-IV categorical personality disorder diagnosis. Our results confirmed the original five-factor structure of the PID-5. The reliability and the convergent and discriminant validity of the PID-5 proved to be adequate. Implications for future research are discussed

The design and construction of a PhET-based inquiry learning worksheet to develop understanding of the particle-in-a-box model

We report on the design and construction of a worksheet to develop upper secondary school students' understanding of the particle-in-a-box model. We designed a worksheet that guided students' structured-inquiry learning through peer discussion using the PhET simulation 'Quantum Bound States'. The worksheet was improved in three iterative cycles of (re)designing, testing and evaluating, leading to a validated design and tentative design principles. Students' discourse was recorded whilst they were using the worksheet and the PhET simulation in the test phase of each cycle. Analyses of students' discourse informed the redesign of the worksheet for each subsequent cycle, until the design was finalised. The results showed the potential of the simulation to introduce upper secondary school students to the particle-in-a-box model, provided care is taken to accompany student inquiry with a well-developed worksheet as learning support during the lesson

Complete genome sequence of the fire blight pathogen Erwinia pyrifoliae DSM 12163T and comparative genomic insights into plant pathogenicity

Smits THM, Jaenicke S, Rezzonico F, et al. Complete genome sequence of the fire blight pathogen Erwinia pyrifoliae DSM 12163(T) and comparative genomic insights into plant pathogenicity. BMC Genomics. 2010;11(1): 2.Background: Erwinia pyrifoliae is a newly described necrotrophic pathogen, which causes fire blight on Asian (Nashi) pear and is geographically restricted to Eastern Asia. Relatively little is known about its genetics compared to the closely related main fire blight pathogen E. amylovora. Results: The genome of the type strain of E. pyrifoliae strain DSM 12163(T), was sequenced using both 454 and Solexa pyrosequencing and annotated. The genome contains a circular chromosome of 4.026 Mb and four small plasmids. Based on their respective role in virulence in E. amylovora or related organisms, we identified several putative virulence factors, including type III and type VI secretion systems and their effectors, flagellar genes, sorbitol metabolism, iron uptake determinants, and quorum-sensing components. A deletion in the rpoS gene covering the most conserved region of the protein was identified which may contribute to the difference in virulence/host-range compared to E. amylovora. Comparative genomics with the pome fruit epiphyte Erwinia tasmaniensis Et1/99 showed that both species are overall highly similar, although specific differences were identified, for example the presence of some phage gene-containing regions and a high number of putative genomic islands containing transposases in the E. pyrifoliae DSM 12163T genome. Conclusions: The E. pyrifoliae genome is an important addition to the published genome of E. tasmaniensis and the unfinished genome of E. amylovora providing a foundation for re-sequencing additional strains that may shed light on the evolution of the host-range and virulence/pathogenicity of this important group of plant-associated bacteria

Bilaterale osteochondritis dissecans van de laterale taluskam bij een jonge rottweiler

In this case report, a young Rottweiler is described that suffered from bilateral osteochondritis dissecans of the talus. The signalment, history and clinical signs were typical for this condition, but atypically, the lateral trochlear ridges were affected. Due to the large size of the fragments, the prognosis was reserved. However, one year after surgery, the clinical outcome was surprisingly good. The left and right trochlear ridges seemed to be filled up with osseous material

Genomics and current genetic understanding of Erwinia amylovora and the fire blight antagonist Pantoea vagans

Abstract The bacterial plant pathogen Erwinia amylovora causes fire blight, a major disease threat to pome fruit production worldwide with further impact on a wide-range of Rosaceae species. Important factors contributing to the development of the disease were discovered in the last decades. Comparative genomics of the genera Erwinia and Pantoea is coming into focus with the recent availability of complete genome sequences. Insights from comparative genomics now position us to answer fundamental questions regarding the evolution of E. amylovora as a successful pathogen and the critical elements for biocontrol activity of Pantoea spp. This trove of new data promises to reveal novel determinants and to understand interactive pathways for virulence, host range and ecological fitness. The ultimate aim is now to apply genomics and identify the pathogen Achilles heels and antagonist mechanisms of action as targets for designing innovative control strategies for fire blight

Quantifying maternal investment in mammals using allometry

Maternal investment influences the survival and reproduction of both mothers and their progeny and plays a crucial role in understanding individuals’ life-history and population ecology. To reveal the complex mechanisms associated with reproduction and investment, it is necessary to examine variations in maternal investment across species. Comparisons across species call for a standardised method to quantify maternal investment, which remained to be developed. This paper addresses this limitation by introducing the maternal investment metric – MI – for mammalian species, established through the allometric scaling of the litter mass at weaning age by the adult mass and investment duration (i.e. gestation + lactation duration) of a species. Using a database encompassing hundreds of mammalian species, we show that the metric is not highly sensitive to the regression method used to fit the allometric relationship or to the proxy used for adult body mass. The comparison of the maternal investment metric between mammalian subclasses and orders reveals strong differences across taxa. For example, our metric confirms that Eutheria have a higher maternal investment than Metatheria. We discuss how further research could use the maternal investment metric as a valuable tool to understand variation in reproductive strategies

Application of the pMHC array to characterise tumour antigen specific T cell populations in leukaemia patients at disease diagnosis

Immunotherapy treatments for cancer are becoming increasingly successful, however to further improve our understanding of the T-cell recognition involved in effective responses and to encourage moves towards the development of personalised treatments for leukaemia immunotherapy, precise antigenic targets in individual patients have been identified. Cellular arrays using peptide-MHC (pMHC) tetramers allow the simultaneous detection of different antigen specific T-cell populations naturally circulating in patients and normal donors. We have developed the pMHC array to detect CD8+ T-cell populations in leukaemia patients that recognise epitopes within viral antigens (cytomegalovirus (CMV) and influenza (Flu)) and leukaemia antigens (including Per Arnt Sim domain 1 (PASD1), MelanA, Wilms’ Tumour (WT1) and tyrosinase). We show that the pMHC array is at least as sensitive as flow cytometry and has the potential to rapidly identify more than 40 specific T-cell populations in a small sample of T-cells (0.8–1.4 x 106). Fourteen of the twenty-six acute myeloid leukaemia (AML) patients analysed had T cells that recognised tumour antigen epitopes, and eight of these recognised PASD1 epitopes. Other tumour epitopes recognised were MelanA (n = 3), tyrosinase (n = 3) and WT1126-134 (n = 1). One of the seven acute lymphocytic leukaemia (ALL) patients analysed had T cells that recognised the MUC1950-958 epitope. In the future the pMHC array may be used provide point of care T-cell analyses, predict patient response to conventional therapy and direct personalised immunotherapy for patients

First geodetic observations using new VLBI stations ASKAP-29 and WARK12M

We report the results of a successful 7 hour 1.4 GHz VLBI experiment using two new stations, ASKAP-29 located in Western Australia and WARK12M located on the North Island of New Zealand. This was the first geodetic VLBI observing session with the participation of these new stations. We have determined the positions of ASKAP-29 and WARK12M. Random errors on position estimates are 150-200 mm for the vertical component and 40-50 mm for the horizontal component. Systematic errors caused by the unmodeled ionosphere path delay may reach 1.3 m for the vertical component.Comment: 11 pages, 6 flgures, 4 table