Phase variation of a Type IIG restriction-modification enzyme alters site-specific methylation patterns and gene expression in Campylobacter jejuni strain NCTC11168

Phase-variable restriction-modification systems are a feature of a diverse range of bacterial species. Stochastic, reversible switches in expression of the methyltransferase produces variation in methylation of specific sequences. Phase-variable methylation by both Type I and Type III methyltransferases is associated with altered gene expression and phenotypic variation. One phase-variable gene of Campylobacter jejuni encodes a homologue of an unusual Type IIG restriction-modification system in which the endonuclease and methyltransferase are encoded by a single gene. Using both inhibition of restriction and PacBio-derived methylome analyses of mutants and phase-variants, the cj0031c allele in C. jejuni strain NCTC11168 was demonstrated to specifically methylate adenine in 5′CCCGA and 5′CCTGA sequences. Alterations in the levels of specific transcripts were detected using RNA-Seq in phase-variants and mutants of cj0031c but these changes did not correlate with observed differences in phenotypic behaviour. Alterations in restriction of phage growth were also associated with phase variation (PV) of cj0031c and correlated with presence of sites in the genomes of these phages. We conclude that PV of a Type IIG restriction-modification system causes changes in site-specific methylation patterns and gene expression patterns that may indirectly change adaptive traits

Limited impact of adolescent meningococcal ACWY vaccination on Neisseria meningitides serogroup W carriage in university students

Background In the UK rising disease levels due to Neisseria meningitidis serogroup W clonal complex ST-11 (MenW:cc11) strains led to introduction of conjugate MenACWY vaccination for teenagers. We investigated the impact of immunization on carriage of targeted meningococci by whole genome sequencing of isolates recovered from a cohort of vaccinated university students. Methods Strain designation data were extracted from whole genome sequence data. Genomes from carried and invasive MenW:cc11 were compared using a gene-by-gene approach. Serogrouping identified isolates expressing capsule antigens targeted by the vaccine. Results Isolates with a W: P1.5,2: F1-1: ST-11 (cc11) designation, and belonging to the emerging ‘2013-strain’ of the South American-UK MenW:cc11 sub-lineage, were responsible for an increase in carried group W. A multifocal expansion was evident with close transmission networks extending beyond individual dormitories. Carried group Y isolates were predominantly from clonal complex 23, but showed significant heterogeneity and individual strain designations were only sporadically recovered. No shifts towards acapsulate phenotypes were detected in targeted meningococcal populations. Conclusions In a setting with high levels of conjugate MenACWY vaccination, expansion of capsule-expressing isolates from the 2013-strain of MenW:cc11, but not MenY:cc23 isolates, is indicative of differential susceptibilities to vaccine-induced immunity

Exploring cancer survivors’ views of health behavior change: "Where do you start, where do you stop with everything?"

International audienceObjective: Physical activity (PA) and a healthy diet can improve the well‐being of cancer survivors. However, cancer survivors often do not engage in these behaviours. This study aimed to explore barriers and facilitators to engaging in these behaviours following cancer treatment.Methods: During the development of a web‐based intervention to enhance health‐related quality of life in cancer survivors, 32 people who had completed treatment for breast, colon or prostate cancer were presented with an intervention for PA and healthy eating. In‐depth think‐aloud and semi‐structured interviewing techniques were used to elicit perceptions of both behaviours. Data were analysed using thematic analysis.Results: Some individuals reported implementing positive health behaviour changes to maintain health and prevent recurrence, or to help them to move forward after cancer. However, others reported feeling abandoned, and many did not report an intention to engage in lifestyle changes. Individuals discussed contextual and health‐related barriers that were specifically linked to their situation as post‐treatment cancer survivors: individuals described uncertainty about how to implement adaptive changes and perceived a lack of support from healthcare providers. Others viewed behaviour change as unnecessary or undesirable, with some arguing that non‐modifiable factors contributed more to their cancer diagnosis than lifestyle‐related factors.Conclusions: For many participants in this study, the period that follows treatment for cancer did not represent a ‘teachable moment’. A variety of complex and heterogeneous factors appeared to impact motivation, and may limit cancer survivors from engaging with diet and PA changes

TESSA: A toolkit for rapid assessment of ecosystem services at sites of biodiversity conservation importance

Sites that are important for biodiversity conservation can also provide significant benefits (i.e. ecosystem services) to people. Decision-makers need to know how change to a site, whether development or restoration, would affect the delivery of services and the distribution of any benefits among stakeholders. However, there are relatively few empirical studies that present this information. One reason is the lack of appropriate methods and tools for ecosystem service assessment that do not require substantial resources or specialist technical knowledge, or rely heavily upon existing data. Here we address this gap by describing the Toolkit for Ecosystem Service Site-based Assessment (TESSA). It guides local non-specialists through a selection of relatively accessible methods for identifying which ecosystem services may be important at a site, and for evaluating the magnitude of benefits that people obtain from them currently, compared with those expected under alternative land-uses. The toolkit recommends use of existing data where appropriate and places emphasis on enabling users to collect new field data at relatively low cost and effort. By using TESSA, the users could also gain valuable information about the alternative land-uses; and data collected in the field could be incorporated into regular monitoring programmes

Computer simulations of developmental change: The contributions of working memory capacity and long-term knowledge

Increasing working memory (WM) capacity is often cited as a major influence on children’s development and yet WM capacity is difficult to examine independently of long-term knowledge. A computational model of children’s nonword repetition (NWR) performance is presented that independently manipulates long-term knowledge and WM capacity to determine the relative contributions of each in explaining the developmental data. The simulations show that (1) both mechanisms independently cause the same overall developmental changes in NWR performance; (2) increase in long-term knowledge provides the better fit to the child data; and (3) varying both long-term knowledge and WM capacity adds no significant gains over varying long-term knowledge alone. Given that increases in long-term knowledge must occur during development, the results indicate that increases in WM capacity may not be required to explain developmental differences. An increase in WM capacity should only be cited as a mechanism of developmental change when there are clear empirical reasons for doing so

Agricultural intensification and the evolution of host specialism in the enteric pathogen Campylobacter jejuni.

Modern agriculture has dramatically changed the distribution of animal species on Earth. Changes to host ecology have a major impact on the microbiota, potentially increasing the risk of zoonotic pathogens being transmitted to humans, but the impact of intensive livestock production on host-associated bacteria has rarely been studied. Here, we use large isolate collections and comparative genomics techniques, linked to phenotype studies, to understand the timescale and genomic adaptations associated with the proliferation of the most common food-born bacterial pathogen (Campylobacter jejuni) in the most prolific agricultural mammal (cattle). Our findings reveal the emergence of cattle specialist C. jejuni lineages from a background of host generalist strains that coincided with the dramatic rise in cattle numbers in the 20th century. Cattle adaptation was associated with horizontal gene transfer and significant gene gain and loss. This may be related to differences in host diet, anatomy, and physiology, leading to the proliferation of globally disseminated cattle specialists of major public health importance. This work highlights how genomic plasticity can allow important zoonotic pathogens to exploit altered niches in the face of anthropogenic change and provides information for mitigating some of the risks posed by modern agricultural systems

Land cover change and carbon emissions over 100 years in an African biodiversity hotspot

Agricultural expansion has resulted in both land use and land cover change (LULCC) across the tropics. However, the spatial and temporal patterns of such change and their resulting impacts are poorly understood, particularly for the pre-satellite era. Here we quantify the LULCC history across the 33.9 million ha watershed of Tanzania's Eastern Arc Mountains, using geo-referenced and digitised historical land cover maps (dated 1908, 1923, 1949 and 2000). Our time series from this biodiversity hotspot shows that forest and savanna area both declined, by 74% (2.8 million ha) and 10% (2.9 million ha), respectively, between 1908 and 2000. This vegetation was replaced by a five-fold increase in cropland, from 1.2 million ha to 6.7 million ha. This LULCC implies a committed release of 0.9 Pg C (95% CI: 0.4-1.5) across the watershed for the same period, equivalent to 0.3 Mg C ha(-1) yr(-1) . This is at least three-fold higher than previous estimates from global models for the same study area. We then used the LULCC data from before and after protected area creation, as well as from areas where no protection was established, to analyse the effectiveness of legal protection on land cover change despite the underlying spatial variation in protected areas. We found that, between 1949 and 2000, forest expanded within legally protected areas, resulting in carbon uptake of 4.8 (3.8-5.7) Mg C ha(-1) , compared to a committed loss of 11.9 (7.2-16.6) Mg C ha(-1) within areas lacking such protection. Furthermore, for nine protected areas where LULCC data is available prior to and following establishment, we show that protection reduces deforestation rates by 150% relative to unprotected portions of the watershed. Our results highlight that considerable LULCC occurred prior to the satellite era, thus other data sources are required to better understand long-term land cover trends in the tropics. This article is protected by copyright. All rights reserved

Restructuring and repositioning of private accomodation in Primorsko-Goranska county: problems and solutions

The paper presents the results of a research conducted on private accommodation as most common form of accommodation within the tourism offer of Primorsko-Goranska County. The study was conducted in 2008, and the obtained results of the research are analysed in the paper, conclusions made, key problems identified and appropriate measures proposed, with the aim of enhancing the quality of this segment of the tourism industry. The goal of the above mentioned measures’ implementation are quality improvement, repositioning and restructuring of private accommodation and its transformation into small family hotels, boarding houses or apart-hotels

Orally bioavailable CDK9/2 inhibitor shows mechanism-based therapeutic potential in MYCN-driven neuroblastoma

The undruggable nature of oncogenic Myc transcription factors poses a therapeutic challenge in neuroblastoma, a pediatric cancer in which MYCN amplification is strongly associated with unfavorable outcome. Here, we show that CYC065 (fadraciclib), a clinical inhibitor of CDK9 and CDK2, selectively targeted MYCN-amplified neuroblastoma via multiple mechanisms. CDK9 — a component of the transcription elongation complex P-TEFb — bound to the MYCN-amplicon superenhancer, and its inhibition resulted in selective loss of nascent MYCN transcription. MYCN loss led to growth arrest, sensitizing cells for apoptosis following CDK2 inhibition. In MYCN-amplified neuroblastoma, MYCN invaded active enhancers, driving a transcriptionally encoded adrenergic gene expression program that was selectively reversed by CYC065. MYCN overexpression in mesenchymal neuroblastoma was sufficient to induce adrenergic identity and sensitize cells to CYC065. CYC065, used together with temozolomide, a reference therapy for relapsed neuroblastoma, caused long-term suppression of neuroblastoma growth in vivo, highlighting the clinical potential of CDK9/2 inhibition in the treatment of MYCN-amplified neuroblastoma