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

Enhanced Retention In The Passive-Avoidance Task By 5-HT1A Receptor Blockade Is Not Associated With Increased Activity Of The Central Nucleus Of The Amygdala

The effect of blockade of S-HT1A receptors was investigated on (1) retention in a mildly aversive passive-avoidance task, and (2) spontaneous single-unit activity of central nucleus of the amygdala (CeA) neurons, a brain site implicated in modulation of retention. Systemic administration of the selective S-HT1A antagonist NAN-190 immediately after training markedly-and dose-dependently-facilitated retention in the passive-avoidance task; enhanced retention was time-dependent and was not attributable to variations in wattages of shock received by animals. Systemic administration of NAN-190 had mixed effects on spontaneous single-unit activity of CeA neurons recorded extracellularly in vivo; microiontophoretic application of S-HT, in contrast, consistently and potently suppressed CeA activity. The present findings-that S-HT1A receptor blockade by NAN-190 (1) enhances retention in the passive-avoidance task, and (2) does not consistently increase spontaneous neuronal activity of the CeA-provide evidence that a serotonergic system tonically inhibits modulation of retention in the passive-avoidance task through activation of the S-HT1A receptor subtype at brain sites located outside the CeA

Diffuse Gastric Ganglioneuromatosis: Novel Presentation of PTEN

Gastrointestinal ganglioneuromatous proliferations are rare, most often found in the colon, and are three types: polypoid ganglioneuromas, ganglioneuromatous polyposis, and diffuse ganglioneuromatosis. We present a case of diffuse ganglioneuromatosis in the posterior gastric wall in a nine-year-old female. To our knowledge, this is the first reported case of diffuse ganglioneuromatosis located in the stomach. Only six cases of gastric ganglioneuromatous proliferations have previously been reported, two in English and none were diffuse ganglioneuromatosis. A diagnosis of diffuse ganglioneuromatosis is relevant for patient care because, unlike sporadic polypoid ganglioneuromas or ganglioneuromatous polyposis, most are syndromic. Diffuse ganglioneuromatosis is commonly associated with neurofibromatosis type 1, multiple endocrine neoplasia type 2b, and Cowden Syndrome, one of the phenotypes of PTEN hamartoma tumor syndrome. The patient had the noted gastric diffuse ganglioneuromatosis, as well as other major and minor criteria for Cowden syndrome. Genetic testing revealed a novel frameshift mutation in the PTEN gene in the patient, her father, paternal aunt, and the aunt’s son who is a paternal first cousin of the patient

Uncovering the natural variability of araucariacean exudates from ex situ and in situ tree populations in New Caledonia using FTIR spectroscopy

Background Understanding the natural variability of plant resins is crucial for evaluating the chemical information stored in ambers that may support inferring palaeoenvironmental conditions. However, even among extant resin-producing plants, the variation of resinous exudates within and between tree genera and species is still poorly understood. Methods We analysed plant exudates from across the Araucariaceae in New Caledonia using Fourier-transform infrared spectroscopy in attenuated total reflection mode (FTIR-ATR). Both, wild (in situ) and arboretum grown (ex situ) populations were used to uncover the effects of intergeneric, inter- and intraspecific variation of the araucariacean plants on the resin chemistry. Results We show that even resins from the same species in the same habitat do have natural (intraspecific) chemical variation. Some of the variation can result from the degree of resin polymerisation, but this is not the only source of variation. Wild sourced resins have greater natural intraspecific chemical variation than the arboretum sourced ones. Interspecific and intergeneric differences were not easy to distinguish in the resins sampled. This has strong implications for the evaluation of the chemical information from worldwide ambers: multiple samples should be analysed to give a more accurate picture of the natural chemical variation present and how this may or may not overlap with the chemistries of other resin types. Additionally we discovered that the Araucariacean species can produce up to three distinct types of exudate (gum, resin, and gum resin). This is the first time that gum resins have been identified and characterized with FTIR-ATR. We also provide a guide on how to distinguish the exudate types when using FTIR-ATR.Peer reviewe

Autodidacticism and Music: Do Self-Taught Musicians Exhibit the Same Auditory Processing Advantages as Formally Trained Musicians?

Multiple studies have demonstrated that musicians have enhanced auditory processing abilities compared to non-musicians. In these studies, musicians are usually defined as having received some sort of formal music training. One issue with this definition is that there are many musicians who are self-taught. The goal of the current study was to determine if self-taught musicians exhibit different auditory enhancements as their formally trained counterparts. Three groups of participants were recruited: formally trained musicians, who received formal music training through the conservatory or private lessons; self-taught musicians, who learned to play music through informal methods, such as with books, videos, or by ear; non-musicians, who had little or no music experience. Auditory processing abilities were assessed using a speech-in-noise task, a passive pitch oddball task done while recording electrical brain activity, and a melodic tonal violation task, done both actively and passively while recording electrical brain activity. For the melodic tonal violation task, formally trained musicians were better at detecting a tonal violation compared to self-taught musicians, who were in turn better than non-musicians. The P600 evoked by a tonal violation was enhanced in formally trained musicians compared to non-musicians. The P600 evoked by an out-of-key note did not differ between formally trained and self-taught musicians, while the P600 evoked by an out-of-tune note was smaller in self-taught musicians compared to formally trained musicians. No differences were observed between the groups for the other tasks. This pattern of results suggests that music training format impacts auditory processing abilities in musical tasks; however, it is possible that these differences arose due to pre-existing factors and not due to the training itself

Functional Brain Hyperactivations Are Linked to an Electrophysiological Measure of Slow Interhemispheric Transfer Time after Pediatric Moderate/Severe Traumatic Brain Injury.

Increased task-related blood oxygen level dependent (BOLD) activation is commonly observed in functional magnetic resonance imaging (fMRI) studies of moderate/severe traumatic brain injury (msTBI), but the functional relevance of these hyperactivations and how they are linked to more direct measures of neuronal function remain largely unknown. Here, we investigated how working memory load (WML)-dependent BOLD activation was related to an electrophysiological measure of interhemispheric transfer time (IHTT) in a sample of 18 msTBI patients and 26 demographically matched controls from the UCLA RAPBI (Recovery after Pediatric Brain Injury) study. In the context of highly similar fMRI task performance, a subgroup of TBI patients with slow IHTT had greater BOLD activation with higher WML than both healthy control children and a subgroup of msTBI patients with normal IHTT. Slower IHTT treated as a continuous variable was also associated with BOLD hyperactivation in the full TBI sample and in controls. Higher WML-dependent BOLD activation was related to better performance on a clinical cognitive performance index, an association that was more pronounced within the patient group with slow IHTT. Our previous work has shown that a subgroup of children with slow IHTT after pediatric msTBI has increased risk for poor white matter organization, long-term neurodegeneration, and poor cognitive outcome. BOLD hyperactivations after msTBI may reflect neuronal compensatory processes supporting higher-order capacity demanding cognitive functions in the context of inefficient neuronal transfer of information. The link between BOLD hyperactivations and slow IHTT adds to the multi-modal validation of this electrophysiological measure as a promising biomarker

The Federal Rules Of Civil Procedure: Politics In The 2013-2014 Revision

Pre-trial discovery is perpetually controversial. Parties advantaged by strict privacy can often avoid justice when this is disadvantageous to their interests. Contrawise, parties advantaged by relaxed litigation privacy can achieve justice when all facts are accessible irrespective of their repositories, ownership or control. American-style pre-trial discovery in civil and regulatory enforcement is relatively rare around the world. U.S. discovery rules open nearly all relevant and non-privileged data for use by opposing parties. The traditional discovery process was costly and time consuming in the world of tangible paper data. However, these burdens have increased, rather than diminished as often predicted, as most data migrates to electronically stored information (ESI). This article provides a midstream assessment of the second major revision effort to accommodate U.S. discovery processes to the broad and deep problems arising during the past 20 years of document discovery experience with predominately ESI data sources

Variation in activity levels amongst dogs of different breeds:results of a large online survey of dog owners from the UK

Regular physical activity is an important means of promoting health, both in people and their pets. Walking is the most common method used for dogs, but there is a lack of clarity on how much daily activity different breeds of dog require. Data from an online survey of UK dog owners were collected between June and August in 2014. The University of Liverpool Ethics Committee approved the project, and owners consented to data use. The initial dataset (17 028 dogs) was first cleaned to remove erroneous data, and then edited to remove mixed breed dogs, leaving a total of 12 314 dogs from known pedigree breeds. Other information collected included sex, age, neuter status, breed, and amount and frequency of exercise. Exercise frequency and duration were estimated across different breeds, and compared with Kennel Club recommendations, using χ 2 tests and binary logistic regression. The online survey data indicated differences amongst breeds in the amount of walking reported (P < 0·001). Afghan hounds were the least exercised breed, whilst breeds reportedly exercised most included: English setter, foxhound, Irish setter and Old English sheepdog. Gundogs were most likely to be walked once per d or more (P < 0·001), whilst smaller dogs were more likely to meet their UK Kennel Club guidelines for dog walking (P < 0·001). The frequency of dog walking varies both within and amongst breeds, and many do not currently receive the recommended amount of exercise. This may constitute a canine welfare problem and also have an impact on the physical activity levels of their owners

Sixty years of Sverdrup : a retrospective of progress in the study of phytoplankton blooms

Author Posting. © The Oceanography Society, 2014. This article is posted here by permission of The Oceanography Society for personal use, not for redistribution. The definitive version was published in Oceanography 27, no. 1 (2014): 222–235, doi:10.5670/oceanog.2014.26.One of the most dramatic large-scale features in the ocean is the seasonal greening of the North Atlantic in spring and summer due to the accumulation of phytoplankton biomass in the surface layer. In 1953, Harald Ulrik Sverdrup hypothesized a now canonical mechanism for the development and timing of phytoplankton blooms in the North Atlantic. Over the next 60 years, Sverdrup's Critical Depth Hypothesis spurred progress in understanding of bloom dynamics and offered a valuable theoretical framework on which to build. In reviewing 60 years of literature, the authors trace the development of modern bloom initiation hypotheses, highlighting three case studies that illuminate the complexity, including both catalysts and impediments, of scientific progress in the wake of Sverdrup's hypothesis. Most notably, these cases demonstrate that the evolution of our understanding of phytoplankton blooms was paced by access not only to technology but also to concurrent insights from several disciplines. This exploration of the trajectories and successes in bloom studies highlights the need for expanding interdisciplinary collaborations to address the complexity of phytoplankton bloom dynamics

Anaerobic Bacteria Grow within Candida albicans Biofilms and Induce Biofilm Formation in Suspension Cultures

The human microbiome contains diverse microorganisms, which share and compete for the same environmental niches. A major microbial growth form in the human body is the biofilm state, where tightly packed bacterial, archaeal, and fungal cells must cooperate and/or compete for resources in order to survive. We examined mixed biofilms composed of the major fungal species of the gut microbiome, Candida albicans, and each of five prevalent bacterial gastrointestinal inhabitants: Bacteroides fragilis, Clostridium perfringens, Escherichia coli, Klebsiella pneumoniae, and Enterococcus faecalis. We observed that biofilms formed by C. albicans provide a hypoxic microenvironment that supports the growth of two anaerobic bacteria, even when cultured in ambient oxic conditions that are normally toxic to the bacteria. We also found that coculture with bacteria in biofilms induces massive gene expression changes in C. albicans, including upregulation of WOR1, which encodes a transcription regulator that controls a phenotypic switch in C. albicans, from the “white” cell type to the “opaque” cell type. Finally, we observed that in suspension cultures, C. perfringens induces aggregation of C. albicans into “mini-biofilms,” which allow C. perfringens cells to survive in a normally toxic environment. This work indicates that bacteria and C. albicans interactions modulate the local chemistry of their environment in multiple ways to create niches favorable to their growth and survival