The role of the phosphopantetheinyltransferase enzyme, PswP, in the biosynthesis of antimicrobial secondary metabolites by <em>Serratia marcescens </em>Db10

Phosphopantetheinyltransferase (PPTase) enzymes fulfil essential roles in primary and secondary metabolism in prokaryotes, archaea and eukaryotes. PPTase enzymes catalyse the essential modification of the carrier protein domain of fatty acid synthases, polyketide synthases (PKSs) and non-ribosomal peptide synthetases (NRPSs). In bacteria and fungi, NRPS and PKS enzymes are often responsible for the biosynthesis of secondary metabolites with clinically relevant properties; these secondary metabolites include a variety of antimicrobial peptides. We have previously shown that in the Gram-negative bacterium Serratia marcescens Db10, the PPTase enzyme PswP is essential for the biosynthesis of an NRPS-PKS dependent antibiotic called althiomycin. In this work we utilize bioinformatic analyses to classify PswP as belonging to the F/KES subfamily of Sfp type PPTases and to putatively identify additional NRPS substrates of PswP, in addition to the althiomycin NRPS-PKS, in Ser. marcescens Db10. We show that PswP is required for the production of three diffusible metabolites by this organism, each possessing antimicrobial activity against Staphylococcus aureus. Genetic analyses identify the three metabolites as althiomycin, serrawettin W2 and an as-yet-uncharacterized siderophore, which may be related to enterobactin. Our results highlight the use of an individual PPTase enzyme in multiple biosynthetic pathways, each contributing to the ability of Ser. marcescens to inhibit competitor bacteria by the production of antimicrobial secondary metabolites

Intersectional inequalities and the U.S. opioid crisis:Challenging dominant narratives and revealing heterogeneities

Dominant narratives of prescription opioid misuse (POM) in the U.S. have portrayed it as an issue primarily affecting White communities. In this study we explore POM as reported in data from the 2015 National Survey on Drug Use and Health, using an intersectional multilevel analysis of individual heterogeneity and discriminatory accuracy (MAIHDA). We map the risk of POM through a series of multilevel models with individuals (N = 43,409) nested within strata formed by the intersections of gender, race/ethnicity, income, and age. We find meaningful heterogeneity between and within strata. The ten strata with the greatest risk for POM were comprised of individuals identifying as White, African American, and non-White Hispanic, and included individuals of low, medium, and high income. We uncover intersections of social position with high risk for POM that are often excluded from dominant narratives, including young high-income African American women. Intersectional approaches are essential for advancing our understanding of health inequalities and unfolding epidemics such as that of POM in the U.S

Dynamiska förmågor bland fyra företag i byggindustrin

Syfte: Syftet med uppsatsen är att undersöka om det i vissa situationer är fördelaktigare för ett företag att förändra sin resursbas med ad hoc-lösningar än genom att utveckla dynamiska förmågor. Metod: Uppsatsen genomförs genom en multipel fallstudie på fyra företag. Analysen av det insamlade materialet utförs med så kallad ”pattern-matching”. Teoretiska perspektiv: Studien använder sig allmänt av det dynamiska perspektivet, och mer specifikt av Teeces (2007) gruppering av dynamiska förmågor i kategorierna förståelse, formning och transformering. Empiri: Insamlingen av data genomfördes med hjälp av semistrukturerade intervjuer med personer på högt uppsatta positioner inom de fyra olika företagen. Resultat: (1) Företagen i undersökningen använder sig av ad hoc-lösningar i högre utsträckning än vad tidigare forskare har antagit är vanligt ute på företag. (2) Det verkar finnas situationer där utvecklandet av dynamiska förmågor är negativt för ett företags lönsamhet. (3) Vår undersökning visar att ett företag eventuellt kan skapa hållbara konkurrensfördelar genom att kontinuerligt optimera sin resursbas med hjälp av ad hoc-lösningar

Image analysis of palm oil crystallisation as observed by hot stage microscopy

This paper was accepted for publication in the journal Journal of Crystal Growth and the definitive published version is available at http://dx.doi.org/10.1016/j.jcrysgro.2016.03.026.An image processing algorithm previously used to analyse the crystallisation of a pure fat (tripalmitin) has been applied to the crystallisation of a multicomponent natural fat (palm oil). In contrast to tripalmitin, which produced circular crystals with a constant growth rate, palm oil produced speckled crystals caused by the inclusion of entrapped liquid, and growth rates gradually decreased with time. This can be explained by the depletion of crystallisable material in the liquid phase, whereas direct impingement of crystals (the basis of the Avrami equation) was less common. A theoretical analysis combining this depletion with assuming that the growth rate is proportional to the supersaturation of a crystallisable pseudo-component predicted a tanh function variation of radius with time. This was generally able to provide good fits to the growth curves. It was found that growth rate was a relatively mild function of temperature but also varied from crystal to crystal and even between different sides of the same crystal, which may be due to variations in composition within the liquid phase. Nucleation rates were confirmed to vary approximately exponentially with decreasing temperature, resulting in much greater numbers of crystals and a smaller final average crystal size at lower temperatures

In vitro replication and thermodynamic studies of methylation and oxidation modifications of 6-thioguanine

The cytotoxic effects of thiopurine drugs are mostly exerted through the formation of thioguanine nucleotide and its subsequent incorporation into DNA. The 6-thioguanine (6-TG) in DNA can be converted to S6-methylthio-2-aminopurine (2-AP-6-SCH3) and 2-aminopurine-6-sulfonic acid (2-AP-6-SO3H) upon reaction with S-adenosyl-L-methionine and irradiation with UVA light, respectively. Here we prepared oligodeoxynucleotides (ODNs) harboring a 6-TG, 2-AP-6-SCH3 or 2-AP-6-SO3H at a defined site and examined, by using LC-MS/MS, the in vitro replication of these substrates with yeast polymerase η and Klenow fragment (KF−). Our results revealed that 2-AP-6-SCH3 could be bypassed by KF−, with significant misincorporation of thymine opposite the lesion. The 2-AP-6-SO3H, however, blocked markedly the nucleotide insertion by KF−. Yeast pol η could bypass all three modified nucleosides; although dCMP was inserted preferentially, we found substantial misincorporation of dTMP and dAMP opposite 2-AP-6-SCH3 and 2-AP-6-SO3H, respectively. Moreover, both KF− and yeast pol η induced a considerable amount of -2 frameshift products from the replication of 2-AP-6-SCH3- and 2-AP-6-SO3H-bearing substrates. Our results also underscored the importance of measuring the relative ionization efficiencies of replication products in the accurate quantification of these products by LC-MS/MS. Moreover, thermodynamic studies revealed that 2-AP-6-SCH3 and 2-AP-6-SO3H could cause more destabilization to duplex DNA than 6-TG. Taken together, the results from this study shed important new light on the biological implications of the two metabolites of 6-TG

A new small regulatory protein, HmuP, modulates haemin acquisition in Sinorhizobium meliloti

Sinorhizobium meliloti has multiple systems for iron acquisition, including the use of haem as an iron source. Haem internalization involves the ShmR haem outer membrane receptor and the hmuTUV locus, which participates in haem transport across the cytoplasmic membrane. Previous studies have demonstrated that expression of the shmR gene is negatively regulated by iron through RirA. Here, we identify hmuP in a genetic screen for mutants that displayed aberrant control of shmR. The normal induction of shmR in response to iron limitation was lost in the hmuP mutant, showing that this gene positively affects shmR expression. Moreover, the HmuP protein is not part of the haemin transporter system. Analysis of gene expression and siderophore production indicates that disruption of hmuP does not affect other genes related to the iron-restriction response. Our results strongly indicate that the main function of HmuP is the transcriptional regulation of shmR. Sequence alignment of HmuP homologues and comparison with the NMR structure of Rhodopseudomonas palustris CGA009 HmuP protein revealed that certain amino acids localized within predicted β-sheets are well conserved. Our data indicate that at least one of the β-sheets is important for HmuP activity

Understanding unequal ageing: towards a synthesis of intersectionality and life course analyses

Intersectionality has received an increasing amount of attention in health inequalities research in recent years. It suggests that treating social characteristics separately—mainly age, gender, ethnicity, and socio-economic position—does not match the reality that people simultaneously embody multiple characteristics and are therefore potentially subject to multiple forms of discrimination. Yet the intersectionality literature has paid very little attention to the nature of ageing or the life course, and gerontology has rarely incorporated insights from intersectionality. In this paper, we aim to illustrate how intersectionality might be synthesised with a life course perspective to deliver novel insights into unequal ageing, especially with respect to health. First we provide an overview of how intersectionality can be used in research on inequality, focusing on intersectional subgroups, discrimination, categorisation, and individual heterogeneity. We cover two key approaches—the use of interaction terms in conventional models and multilevel models which are particularly focussed on granular subgroup differences. In advancing a conceptual dialogue with the life course perspective, we discuss the concepts of roles, life stages, transitions, age/cohort, cumulative disadvantage/advantage, and trajectories. We conclude that the synergies between intersectionality and the life course hold exciting opportunities to bring new insights to unequal ageing and its attendant health inequalities

Scientific Opinion addressing the state of the science on risk assessment of plant protection products for in-soil organisms

Following a request from EFSA, the Panel on Plant Protection Products and their Residues developed an opinion on the science behind the risk assessment of plant protection products for in-soil organisms. The current risk assessment scheme is reviewed, taking into account new regulatory frameworks and scientific developments. Proposals are made for specific protection goals for in-soil organisms being key drivers for relevant ecosystem services in agricultural landscapes such as nutrient cycling, soil structure, pest control and biodiversity. Considering the time-scales and biological processes related to the dispersal of the majority of in-soil organisms compared to terrestrial non-target arthropods living above soil, the Panel proposes that in-soil environmental risk assessments are made at in- and off-field scale considering field boundary levels. A new testing strategy which takes into account the relevant exposure routes for in-soil organisms and the potential direct and indirect effects is proposed. In order to address species recovery and long-term impacts of PPPs, the use of population models is also proposed