New insights into the regulatory pathways associated with the activation of the stringent response in bacterial resistance to the PBP2-targeted antibiotics, mecillinam and OP0595/RG6080

Objectives: The diazabicyclooctane β-lactamase inhibitor OP0595 (RG6080) also acts as an antibiotic, targeting PBP2 in Enterobacteriaceae, but this activity is vulnerable to mutational resistance. We used WGS to investigate the basis of this resistance. Methods: Twenty OP0595-selected mutants, comprising four derived from each of five different Escherichia coli strains, were sequenced on Illumina HiSeq. Reads from each mutant were mapped to the assembled genome of the corresponding parent. A variant-calling file generated with Samtools was parsed to determine genetic alterations. Results: Besides OP0595, the mutants consistently showed decreased susceptibility to mecillinam, which likewise targets PBP2, and grew as stable round forms in the presence of subinhibitory concentrations of OP0595. Among the 20 mutants, 18 had alterations in genes encoding tRNA synthase and modification functions liable to induce expression of the RpoS sigma factor through activation of the stringent response or had mutations suppressing inactivators of RpoS or the stringent response signal-degrading enzyme, SpoT. TolB was inactivated in one mutant: this activates RcsBC regulation and was previously associated with mecillinam resistance. The mechanism of resistance remained unidentified in one mutant. Both the RpoS and RcsBC systems regulate genes of cell division, including ftsAQZ that can compensate for loss or inhibition of PBP2, allowing survival of the challenged bacteria as stable round forms, as seen. Conclusions: WGS identified the global stringent response signal, entailing induction of RpoS, as the main mediator of mutational resistance to OP0595 in E. coli

HST followup observations of two bright z ~ 8 candidate galaxies from the BoRG pure-parallel survey

We present followup imaging of two bright (L > L*) galaxy candidates at z > 8 from the Brightest of Reionizing Galaxies (BoRG) survey with the F098M filter on HST/WFC3. The F098M filter provides an additional constraint on the flux blueward of the spectral break, and the observations are designed to discriminate between low- and high-z photometric redshift solutions for these galaxies. Our results confirm one galaxy, BoRG 0116+1425 747, as a highly probable z ~ 8 source, but reveal that BoRG 0116+1425 630 - previously the brightest known z > 8 candidate (mAB = 24.5) - is likely to be a z ~ 2 interloper. As this source was substantially brighter than any other z > 8 candidate, removing it from the sample has a significant impact on the derived UV luminosity function in this epoch. We show that while previous BoRG results favored a shallow power-law decline in the bright end of the luminosity function prior to reionization, there is now no evidence for departure from a Schechter function form and therefore no evidence for a difference in galaxy formation processes before and after reionization.Comment: Accepted by ApJL, 7 pages, 4 figure

The bright-end galaxy candidates at z ~ 9 from 79 independent HST fields

We present a full data analysis of the pure-parallel Hubble Space Telescope (HST) imaging observations in the Brightest of Reionizing Galaxies Survey (BoRG[z9]) in Cycle 22. The medium-deep exposures with five HST/WFC3IR+UVIS filter bands from 79 independent sightlines (~370 arcmin^2) provide the least biased determination of number density for z>9 bright galaxies against cosmic variance. After a strict two-step selection for candidate galaxies, including dropout color and photometric redshift analyses, and revision of previous BoRG candidates, we identify one source at z~10 and two sources at z~9. The z~10 candidate shows evidence of line-of-sight lens magnification (mu~1.5), yet it appears surprisingly luminous (MUV ~ -22.6\pm0.3 mag), making it one of the brightest candidates at z > 8 known (~ 0.3 mag brighter than the z = 8.68 galaxy EGSY8p7, spectroscopically confirmed by Zitrin and collaborators). For z ~ 9 candidates, we include previous data points at fainter magnitudes and find that the data are well fitted by a Schechter luminosity function with alpha ~ -2.1, MUV ~ -21.5 mag, and log phi ~ -4.5 Mpc^-3mag^-1, for the first time without fixing any parameters. The inferred cosmic star formation rate density is consistent with unaccelerated evolution from lower redshift.Comment: 18pages, 7figures, 6tables. accepted to the Astrophysical Journa

A randomised controlled trial of PEGASUS, a psychoeducational programme for young people with high-functioning autism spectrum disorder.

Psychoeducation is an essential component of postdiagnostic care for people with ASD (autism spectrum disorder), but there is currently no evidence base for clinical practice. We designed, manualised and evaluated PEGASUS (psychoeducation group for autism spectrum understanding and support), a group psychoeducational programme aiming to enhance the self-awareness of young people with ASD by teaching them about their diagnosis

Evaluation of multiplex tandem PCR (MT-PCR) assays for the detection of bacterial resistance genes among Enterobacteriaceae in clinical urines

Background: Increasing resistance drives empirical use of less potent and previously reserved antibiotics, including for urinary tract infections (UTIs). Molecular profiling, without culture, might better guide early therapy.  Objectives: To explore the potential of AusDiagnostics multiplex tandem (MT) PCR UTI assays.  Methods: Two MT-PCR assays were developed successively, seeking 8 or 16 resistance genes. Amplification was tracked in real time, with melting temperatures used to confirm product identity. Assays were variously performed on: (i) extracted DNA; (ii) cultured bacteria; (iii) urine spiked with reference strains; and (iv) bacteria harvested from clinical urines. Results were compared with those from sequencing, real-time SybrGreen PCR or phenotypic susceptibility.  Results: Performance was similar irrespective of whether DNA, cultures or urines were used, with >90% sensitivity and specificity with respect to common β-lactamases, dfr genes and aminoglycoside resistance determinants except aadA1/A2/A3, for which carriage correlated poorly with streptomycin resistance. Fluoroquinolone-susceptible and -resistant Escherichia coli (but not other species) were distinguished by the melting temperatures of their gyrA PCR products. The time from urine to results was <3 h.  Conclusions: The MT-PCR assays rapidly identified resistance genes from Gram-negative bacteria in urines as well as from cultivated bacteria. Used directly on urines, this assay has the potential to guide early therapy

Carbapenem-non-susceptible Enterobacteriaceae in Europe : conclusions from a meeting of national experts

The emergence and global spread of carbapenemase-producing Enterobacteriaceae is of great concern to health services worldwide. These bacteria are often resistant to all beta-lactam antibiotics and frequently co-resistant to most other antibiotics, leaving very few treatment options. The epidemiology is compounded by the diversity of carbapenem-hydrolysing enzymes and the ability of their genes to spread between different bacterial species. Difficulties are also encountered by laboratories when trying to detect carbapenemase production during routine diagnostic procedures due to an often heterogeneous expression of resistance. Some of the resistance genes are associated with successful clonal lineages which have a selective advantage in those hospitals where antimicrobial use is high and opportunities for transmission exist; others are more often associated with transmissible plasmids. A genetically distinct strain of Klebsiella pneumoniae sequence type (ST) 258 harbouring the K. Pneumoniae carbapenemases (KPC) has been causing epidemics of national and international proportions. It follows the pathways of patient referrals, causing hospital outbreaks along the way. Simultaneously, diverse strains harbouring New Delhi metallo-beta-lactamase (NDM-1) are repeatedly being imported into Europe, commonly via patients with prior medical exposure in the Indian subcontinent. Since the nature and scale of carbapenem-non-susceptible Entrobacteriaceae as a threat to hospital patients in Europe remains unclear, a consultation of experts from 31 countries set out to identify the gaps in diagnostic and response capacity, to index the magnitude of carbapenem-non-susceptibility across Europe using a novel five-level staging system, and to provide elements of a strategy to combat this public health issue in a concerted manner.peer-reviewe

Re: "Comparison of antipseudomonal betalactams for febrile neutropenia empiric therapy: systematic review and network metaanalysis" by Horita et al

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