Differential DNA accessibility to polymerase enables 30-minute phenotypic β-lactam antibiotic susceptibility testing of carbapenem-resistant Enterobacteriaceae

The rise in carbapenem-resistant Enterobacteriaceae (CRE) infections has created a global health emergency, underlining the critical need to develop faster diagnostics to treat swiftly and correctly. Although rapid pathogen-identification (ID) tests are being developed, gold-standard antibiotic susceptibility testing (AST) remains unacceptably slow (1–2 d), and innovative approaches for rapid phenotypic ASTs for CREs are urgently needed. Motivated by this need, in this manuscript we tested the hypothesis that upon treatment with β-lactam antibiotics, susceptible Enterobacteriaceae isolates would become sufficiently permeabilized, making some of their DNA accessible to added polymerase and primers. Further, we hypothesized that this accessible DNA would be detectable directly by isothermal amplification methods that do not fully lyse bacterial cells. We build on these results to develop the polymerase-accessibility AST (pol-aAST), a new phenotypic approach for β-lactams, the major antibiotic class for gram-negative infections. We test isolates of the 3 causative pathogens of CRE infections using ceftriaxone (CRO), ertapenem (ETP), and meropenem (MEM) and demonstrate agreement with gold-standard AST. Importantly, pol-aAST correctly categorized resistant isolates that are undetectable by current genotypic methods (negative for β-lactamase genes or lacking predictive genotypes). We also test contrived and clinical urine samples. We show that the pol-aAST can be performed in 30 min sample-to-answer using contrived urine samples and has the potential to be performed directly on clinical urine specimens

Terminology for reporting nitrate concentration (1993)

Reviewed October 199

Resources and student achievement – evidence from a Swedish policy reform

This paper utilizes a policy change to estimate the effect of teacher density on student performance. We find that an increase in teacher density has a positive effect on student achievement. The baseline estimate – obtained by using the grade point average as the outcome variable – implies that resource increases corresponding to the class-size reduction in the STAR-experiment (i.e., a reduction of 7 students) improves performance by 2.6 percentile ranks (or 0.08 standard deviations). When we use test score data for men, potentially a more objective measure of student performance, the effect of resources appears to be twice the size of the baseline estimate.Student performance; teacher/student ratio; policy reform; differences-in-differences

On defining the Hamiltonian beyond quantum theory

Energy is a crucial concept within classical and quantum physics. An essential tool to quantify energy is the Hamiltonian. Here, we consider how to define a Hamiltonian in general probabilistic theories, a framework in which quantum theory is a special case. We list desiderata which the definition should meet. For 3-dimensional systems, we provide a fully-defined recipe which satisfies these desiderata. We discuss the higher dimensional case where some freedom of choice is left remaining. We apply the definition to example toy theories, and discuss how the quantum notion of time evolution as a phase between energy eigenstates generalises to other theories.Comment: Authors' accepted manuscript for inclusion in the Foundations of Physics topical collection on Foundational Aspects of Quantum Informatio

The geometry of the light-cone cell decomposition of moduli space

S.R. is supported by STFC consolidated Grant No. ST/L000415/1 “String Theory, Gauge Theory and Dualit

Using NDF and ADF to balance diets (1990)

In University of Missouri Agricultural guides G3150 and G3160 we explained how detergent solutions are used to measure forage fiber. The guides show how neutral detergent solution can be used to measure neutral detergent fiber (NDF). NDF represents the total plant fiber or cell wall including hemicellulose, cellulose and lignin. These guides also show how acid detergent solution can be used to measure acid detergent fiber (ADF), which contains cellulose and lignin. Both ADF and NDF data help to more accurately estimate feed intake, energy values and animal performanceNew7/90f7M

Animal waste management

"71/1M""Livestock producers have asked for guidelines on animal waste management that will be feasible and enduring. The Missouri Water Pollution Board has been aware of the need for improvements in methods of handling waste from confined feeding operations and for guidelines for producers. Chapter 204 of Missouri Statutes, as amended, gives the Water Pollution Board the responsibility and authority to correct and/or prevent "pollution" of "waters of the state." These terms are defined in the law and discussed briefly in the first section. With these facts in mind, staff engineers of the Water Pollution Board held a series of meetings with staff members of the Extension Division and Department of Agricultural Engineering of the University of Missouri-Columbia to develop guidelines for disposing of waste from confinement feeding operations. This report is a result of their combined efforts. Others assisting with various phases of development of these guidelines included: School of Engineering, University of Missouri-Columbia; State Department of Health, and the Soil Conservation Service. Research data and experience in handling livestock wastes have been used to develop the guidelines for planning, design, construction, and management of alternative systems of livestock waste management. The information and design guidelines herein are intended primarily for the use of personnel in agencies concerned with animal waste management problems." --PrefaceMissouri Water Pollution Board and Extension Division, University of Missouri - Columbia

Pasts and pagan practices: moving beyond Stonehenge

Theorizing the past is not restricted to archaeology and interpretations of 'past' both influence and are themselves constituted within politicized understandings of self, community and in certain instances, spirituality. 'The past in the imagination of the present' is appropriated, variously, to give meaning to the present or to justify actions and interpret experiences. Summer solstice at Stonehenge, with an estimated 21,000 celebrants in 2005, is only the most publicized appropriation (by pagans and other adherents of alternative spirituality and partying) of a 'sacred site'; and conflicts and negotiations occurring throughout Britain are represented in popular and academic presentations of this 'icon of Britishness'. This paper presents work from the Sacred Sites, Contested Rites/Rights Project (http://www.sacredsites.org.uk) project, a collaboration of archaeology and anthropology informed by pagan and alternative approaches and standpoints investigating and theorizing discourse and practice of heritage management and pagan site users. Whether in negotiations around the Stonehenge solstice access or in dealing with numerous other sites, boundaries between groups or discourses are not clearly drawn - discursive communities merge and re-emerge. But clearly 'past' and 'site' are increasingly important within today's Britain, even as television archaeology increases its following, and pagan numbers continue to grow.</p

Differential DNA accessibility to polymerase enables 30-minute phenotypic β-lactam antibiotic susceptibility testing of carbapenem-resistant Enterobacteriaceae

The rise in carbapenem-resistant Enterobacteriaceae (CRE) infections has created a global health emergency, underlining the critical need to develop faster diagnostics to treat swiftly and correctly. Although rapid pathogen-identification (ID) tests are being developed, gold-standard antibiotic susceptibility testing (AST) remains unacceptably slow (1–2 d), and innovative approaches for rapid phenotypic ASTs for CREs are urgently needed. Motivated by this need, in this manuscript we tested the hypothesis that upon treatment with β-lactam antibiotics, susceptible Enterobacteriaceae isolates would become sufficiently permeabilized, making some of their DNA accessible to added polymerase and primers. Further, we hypothesized that this accessible DNA would be detectable directly by isothermal amplification methods that do not fully lyse bacterial cells. We build on these results to develop the polymerase-accessibility AST (pol-aAST), a new phenotypic approach for β-lactams, the major antibiotic class for gram-negative infections. We test isolates of the 3 causative pathogens of CRE infections using ceftriaxone (CRO), ertapenem (ETP), and meropenem (MEM) and demonstrate agreement with gold-standard AST. Importantly, pol-aAST correctly categorized resistant isolates that are undetectable by current genotypic methods (negative for β-lactamase genes or lacking predictive genotypes). We also test contrived and clinical urine samples. We show that the pol-aAST can be performed in 30 min sample-to-answer using contrived urine samples and has the potential to be performed directly on clinical urine specimens

Human mass balance study of the novel anticancer agent ixabepilone using accelerator mass spectrometry

Ixabepilone (BMS-247550) is a semi-synthetic, microtubule stabilizing epothilone B analogue which is more potent than taxanes and has displayed activity in taxane-resistant patients. The human plasma pharmacokinetics of ixabepilone have been described. However, the excretory pathways and contribution of metabolism to ixabepilone elimination have not been determined. To investigate the elimination pathways of ixabepilone we initiated a mass balance study in cancer patients. Due to autoradiolysis, ixabepilone proved to be very unstable when labeled with conventional [14C]-levels (100 μCi in a typical human radio-tracer study). This necessitated the use of much lower levels of [14C]-labeling and an ultra-sensitive detection method, Accelerator Mass Spectrometry (AMS). Eight patients with advanced cancer (3 males, 5 females; median age 54.5 y; performance status 0–2) received an intravenous dose of 70 mg, 80 nCi of [14C]ixabepilone over 3 h. Plasma, urine and faeces were collected up to 7 days after administration and total radioactivity (TRA) was determined using AMS. Ixabepilone in plasma and urine was quantitated using a validated LC-MS/MS method. Mean recovery of ixabepilone-derived radioactivity was 77.3% of dose. Fecal excretion was 52.2% and urinary excretion was 25.1%. Only a minor part of TRA is accounted for by unchanged ixabepilone in both plasma and urine, which indicates that metabolism is a major elimination mechanism for this drug. Future studies should focus on structural elucidation of ixabepilone metabolites and characterization of their activities