Zinc and manganese chelation by neutrophil S100A8/A9 (Calprotectin) limits extracellular aspergillus fumigatus hyphal growth and corneal infection

PMCID: PMC4684987.-- et al.Calprotectin, a heterodimer of S100A8 and S100A9, is an abundant neutrophil protein that possesses antimicrobial activity primarily because of its ability to chelate zinc and manganese. In the current study, we showed that neutrophils from calprotectindeficient S100A9 mice have an impaired ability to inhibit Aspergillus fumigatus hyphal growth in vitro and in infected corneas in a murine model of fungal keratitis; however, the ability to inhibit hyphal growth was restored in S100A9 mice by injecting recombinant calprotectin. Furthermore, using recombinant calprotectin with mutations in either the Zn and Mn binding sites or the Mn binding site alone, we show that both zinc and manganese binding are necessary for calprotectin's antihyphal activity. In contrast to hyphæ, we found no role for neutrophil calprotectin in uptake or killing of intracellular A. fumigatus conidia either in vitro or in a murine model of pulmonary aspergillosis. We also found that an A. fumigatus δzafA mutant, which demonstrates deficient zinc transport, exhibits impaired growth in infected corneas and following incubation with neutrophils or calprotectin in vitro as compared with wild-type. Collectively, these studies demonstrate a novel stage-specific susceptibility of A. fumigatus to zinc and manganese chelation by neutrophil-derived calprotectin.This work was supported by National Eye Institute Grant R01 EY18612 (to E.P.), Visual Science Research Center Core Grant P30EY011373 (to Case Western Reserve University), National Institute of Allergy and Infectious Diseases Grant R01 AI101171 (to E.P.S. and W.J.C.), National Institute of Allergy and Infectious Diseases Immunology Training Grant 5T32AI089474 (to H.L.C.), National Eye Institute Visual Science Training Grant T32 EY007157 (to H.L.C.), and National Eye Institute National Research Service Award Grant 1F30EY025548-01 (to H.L.C.). T.M.H. and A.J. received Lucille Castori Center for Microbes, Inflammation, and Cancer Grants and National Institute of Allergy and Infectious Diseases Grants R01 AI093808 and R21 AI105617 (to T.M.H.). T.M.H. is an investigator in the Pathogenesis of Infectious Diseases supported by the Burroughs Wellcome Fund. J.A.C. was supported by the Spanish Ministry of Economy and Competitiveness through Grant SAF2013-48382-R.Peer Reviewe

Metacognitions and Misappraisal of Cravings in Current and Former Smokers

Models of obsessive-compulsive disorder (OCD) posit that appraisals of intrusive thoughts as highly personally meaningful drive the performance of compulsions and rituals to alleviate the anxiety associated with these thoughts. Recently, catastrophic and dysfunctional appraisals of cravings have been hypothesized to drive smoking behaviors in smokers currently engaged in a cessation effort (Nosen & Woody, 2009). Indeed, certain anxiety-related factors such as anxiety sensitivity (AS) and misappraisals of cravings have been found to predict relapse (Assayag, Bernstein, Zvolenksy, Steeves, & Stewart, 2012; Johnson, Stewart, Steeves, & Zvolensky, 2012; Nosen & Woody, 2009). However, the differences between current and former smokers with regards to AS and craving appraisals have yet to be examined. The purpose of the present study is to investigate the metacognitive differences between current and former smokers. Ninety current and former (recent quitters and long-term quitters) smokers completed measures of dysfunctional metacognitions, craving appraisals, obsessive-compulsive smoking behaviors, anxiety sensitivity, and negative affect. Current smokers scored higher than long-term quitters on many of these constructs, while mixed results were observed for the differences between recent quitters and current smokers, and recent quitters and long-term quitters. Negative affect was found to account for many of these differences, but did not account for the differences between current smokers and recent quitters, and between current smokers and long-term quitters, with regards to obsessive-compulsive smoking. The differences between recent quitters and long-term quitters on beliefs about responsibility for harm and perfectionism/intolerance of uncertainty also remained significant after controlling for negative affect.Bachelor of Scienc

Prescribing for Children With Rheumatic Disease: Perceived Treatment Approaches Between Pediatric and Adult Rheumatologists

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141687/1/acr23273.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/141687/2/acr23273_am.pd

Collision Induced Dissociation Products of Disulfide-bonded Peptides: Ions Result from the Cleavage of More than One Bond

This document is the Accepted Manuscript version of a Published Work that appeared in final form in the Journal of the American Chemical Society, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://doi.org/10.1007/s13361-010-0064-x.Disulfide bonds are a posttranslational modification (PTM) that can be scrambled or shuffled to non-native bonds during recombinant expression, sample handling, or sample purification. Currently, mapping of disulfide bonds is difficult due, to various sample requirements and data analysis difficulties. One step towards facilitating this difficult work is developing a better understanding of how disulfide-bonded peptides fragment during Collision Induced Dissociation (CID). Most automated analysis algorithms function based on the assumption that the preponderance of product ions observed during the dissociation of disulfide-bonded peptides result from the cleavage of just one peptide bond, and in this report we tested that assumption by extensively analyzing the product ions generated when several disulfide-bonded peptides are subjected to CID on a QTOF instrument. We found that one of the most common types of product ions generated resulted from two peptide bond cleavages, or a double cleavage. We found that for several of the disulfide-bonded peptides analyzed, the number of double cleavage product ions outnumbered those of single cleavages. The influence of charge state and precursor ion size was investigated, to determine if those parameters dictated the amount of double cleavage product ions formed. It was found in this sample set that no strong correlation existed between the charge state or peptide size and the portion of product ions assigned as double cleavages. This data shows that these ions could account for many of the product ions detected in CID data of disulfide bonded peptides. We also showed the utility of double cleavage product ions on a peptide with multiple cysteines present. Double cleavage products were able to fully characterize the bonding pattern of each cysteine where typical single b/y cleavage products could not

Assessing the relative validity of the Scottish Collaborative Group FFQ for measuring dietary intake in adults

Acknowledgements: The authors would like to thank Jacqueline Burr and Lindsey Shaw for collecting the data for this study. Data coding and entry for the food diaries was completed by Dr Lindsey Masson. The authors would also like to acknowledge the Scottish Health Survey Team, the Scottish Government and the National Centre for Social Research for their support in conducting this research. Financial support: This work was supported by funding from the Rural and Environment Science and Analytical Services Division (RESAS) programme of the Scottish Government (J.L.H., L.C.A.C., S.W. and G.Mc.N.). The RESAS programme had no role in the design, analysis or writing of this article. Conflict of interest: None. Authorship: J.L.H., L.C.A.C., S.W. and G.Mc.N. were responsible for the design of the study and formulated the research question. L.C.A.C. and S.W. carried out the study. J.L.H. completed the literature review, conducted the statistical analysis and drafted the initial paper. All authors were responsible for drafting and revising the manuscript and have approved the final version. Ethics of human subject participation: This study was conducted according to the guidelines laid down in the Declaration of Helsinki and all procedures involving human subjects were approved by the Rowett Human Studies Ethical Review Panel. Written informed consent was obtained from all participants.Peer reviewedPostprintPostprintPostprintPostprintPostprintPostprintPostprintPostprin

Reducing Stress after Trauma (ReSeT): Study Protocol for a Randomized, Controlled Trial of an Online Psychoeducational Program and Video Therapy Sessions for Children Hospitalized after Trauma

BACKGROUND: Post-traumatic stress symptoms develop in a quarter to half of injured children affecting their longer-term psychologic and physical health. Evidence-based care exists for post-traumatic stress; however, it is not readily available in some communities. We have developed an eHealth program consisting of online, interactive educational modules and telehealth therapist support based in trauma-focused cognitive behavioral therapy, the Reducing Stress after Trauma (ReSeT) program. We hypothesize that children with post-traumatic stress who participate in ReSeT will have fewer symptoms compared to the usual care control group. METHODS: This is a randomized controlled trial to test the effectiveness of the ReSeT intervention in reducing symptoms of post-traumatic stress compared to a usual care control group. One hundred and six children ages 8-17 years, who were admitted to hospital following an injury, with post-traumatic stress symptoms at 4 weeks post-injury, will be recruited and randomized from the four participating trauma centers. The outcomes compared across groups will be post-traumatic stress symptoms at 10 weeks (primary outcome) controlling for baseline symptoms and at 6 months post-randomization (secondary outcome). DISCUSSION: ReSeT is an evidence-based program designed to reduce post-traumatic stress symptoms among injured children using an eHealth platform. Currently, the American College of Surgeons standards suggest that trauma programs identify and treat patients at high risk for mental health needs in the trauma system. If effectiveness is demonstrated, ReSeT could help increase access to evidence-based care for children with post-traumatic stress within the trauma system

Correlates of Complete Childhood Vaccination in East African Countries.

Despite the benefits of childhood vaccinations, vaccination rates in low-income countries (LICs) vary widely. Increasing coverage of vaccines to 90% in the poorest countries over the next 10 years has been estimated to prevent 426 million cases of illness and avert nearly 6.4 million childhood deaths worldwide. Consequently, we sought to provide a comprehensive examination of contemporary vaccination patterns in East Africa and to identify common and country-specific barriers to complete childhood vaccination. Using data from the Demographic and Health Surveys (DHS) for Burundi, Ethiopia, Kenya, Rwanda, Tanzania, and Uganda, we looked at the prevalence of complete vaccination for polio, measles, Bacillus Calmette-Guérin (BCG) and DTwPHibHep (DTP) as recommended by the WHO among children ages 12 to 23 months. We conducted multivariable logistic regression within each country to estimate associations between complete vaccination status and health care access and sociodemographic variables using backwards stepwise regression. Vaccination varied significantly by country. In all countries, the majority of children received at least one dose of a WHO recommended vaccine; however, in Ethiopia, Tanzania, and Uganda less than 50% of children received a complete schedule of recommended vaccines. Being delivered in a public or private institution compared with being delivered at home was associated with increased odds of complete vaccination status. Sociodemographic covariates were not consistently associated with complete vaccination status across countries. Although no consistent set of predictors accounted for complete vaccination status, we observed differences based on region and the location of delivery. These differences point to the need to examine the historical, political, and economic context of each country in order to maximize vaccination coverage. Vaccination against these childhood diseases is a critical step towards reaching the Millennium Development Goal of reducing under-five mortality by two-thirds by 2015 and thus should be a global priority

RAD sequencing resolves fine-scale population structure in a benthic invertebrate: implications for understanding phenotypic plasticity

The field of molecular ecology is transitioning from the use of small panels of classical genetic markers such as microsatellites to much larger panels of single nucleotide polymorphisms (SNPs) generated by approaches like RAD sequencing. However, few empirical studies have directly compared the ability of these methods to resolve population structure. This could have implications for understanding phenotypic plasticity, as many previous studies of natural populations may have lacked the power to detect genetic differences, especially over micro-geographic scales. We therefore compared the ability of microsatellites and RAD sequencing to resolve fine-scale population structure in a commercially important benthic invertebrate by genotyping great scallops (Pecten maximus) from nine populations around Northern Ireland at 13 microsatellites and 10 539 SNPs. The shells were then subjected to morphometric and colour analysis in order to compare patterns of phenotypic and genetic variation. We found that RAD sequencing was superior at resolving population structure, yielding higher Fst values and support for two distinct genetic clusters, whereas only one cluster could be detected in a Bayesian analysis of the microsatellite dataset. Furthermore, appreciable phenotypic variation was observed in size-independent shell shape and coloration, including among localities that could not be distinguished from one another genetically, providing support for the notion that these traits are phenotypically plastic. Taken together, our results suggest that RAD sequencing is a powerful approach for studying population structure and phenotypic plasticity in natural populations

Metal worker’s lung: spatial association with Mycobacterium avium

Background Outbreaks of hypersensitivity pneumonitis (HP) are not uncommon in workplaces where metal working fluid (MWF) is used to facilitate metal turning. Inhalation of microbe-contaminated MWF has been assumed to be the cause, but previous investigations have failed to establish a spatial relationship between a contaminated source and an outbreak. Objectives After an outbreak of five cases of HP in a UK factory, we carried out blinded, molecular-based microbiological investigation of MWF samples in order to identify potential links between specific microbial taxa and machines in the outbreak zone. Methods Custom-quantitative PCR assays, microscopy and phylogenetic analyses were performed on blinded MWF samples to quantify microbial burden and identify potential aetiological agents of HP in metal workers. Measurements and main results MWF from machines fed by a central sump, but not those with an isolated supply, was contaminated by mycobacteria. The factory sump and a single linked machine at the centre of the outbreak zone, known to be the workstation of the index cases, had very high levels of detectable organisms. Phylogenetic placement of mycobacterial taxonomic marker genes generated from these samples indicated that the contaminating organisms were closely related to Mycobacterium avium. Conclusions We describe, for the first time, a close spatial relationship between the abundance of a mycobacterium-like organism, most probably M. avium, and a localised outbreak of MWF-associated HP. The further development of sequence-based analytic techniques should assist in the prevention of this important occupational disease

What behavior change techniques are associated with effective interventions to reduce screen time in 0-5 year olds? A narrative systematic review.

Screen time has been linked to obesity in young children. Therefore, this systematic review aims to investigate which Behavior Change Techniques (BCTs) are associated with the effectiveness of interventions to reduce screen time in 0-5 year olds. Seven databases were searched, including PsycInfo, PubMed, and Medline. Grey literature searches were conducted. Inclusion criteria were interventions reporting pre- and post- outcomes with the primary objective of reducing screen time in 0-5 year olds. Studies were quality assessed using the Effective Public Health Practice Project criteria. Data extracted included participant characteristics, intervention characteristics and screen time outcomes. The BCT Taxonomy was used to extract BCTs. Interventions were categorised as “very”, “quite” or “non” promising based on effect sizes. BCTs were deemed promising if they were in twice as many very/quite promising interventions as non-promising interventions. Seven randomised controlled trials were included, involving 642 participants between 2.5-5.0 years old. One very promising, four quite promising, and two non-promising interventions were identified. Screen time decreased by 25-39 minutes per day in very/quite promising interventions. Eleven BCTs were deemed promising, including “behavior substitution” and “information about social and environmental consequences”. This review identified eleven promising BCTs, which should be incorporated into future screen time interventions with young children. However, most included studies were of weak quality and limited by the populations targeted. Therefore, future methodologically rigorous interventions targeting at-risk populations with higher screen time, such as those of a low socioeconomic status and children with a high BMI, should be prioritized