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We perform a deep survey of planetary nebulae (PNe) in the spiral galaxy NGC 300 to construct its planetary nebula luminosity function (PNLF). We aim to derive the distance using the PNLF and to probe the characteristics of the most luminous PNe. We analyse 44 fields observed with MUSE at the VLT, covering a total area of $\sim11$ kpc$^2$. We find [OIII]5007 sources using the differential emission line filter (DELF) technique. We identify PNe through spectral classification using the aid of the BPT-diagram. The PNLF distance is derived using the maximum likelihood estimation technique. For the more luminous PNe, we also measure their extinction using the Balmer decrement. We estimate the luminosity and effective temperature of the central stars of the luminous PNe, based on estimates of the excitation class and the assumption of optically thick nebulae. We identify 107 PNe and derive a most-likely distance modulus $(m-M)_0 = 26.48^{+0.11}_{-0.26}$ ($d = 1.98^{+0.10}_{-0.23}$ Mpc). We find that the PNe at the PNLF cut-off exhibit relatively low extinction, with some high extinction cases caused by local dust lanes. We present the lower limit luminosities and effective temperatures of the central stars for some of the brighter PNe. We also identify a few Type I PNe that come from a young population with progenitor masses $>2.5 \, M_\odot$, however do not populate the PNLF cut-off. The spatial resolution and spectral information of MUSE allow precise PN classification and photometry. These capabilities also enable us to resolve possible contamination by diffuse gas and dust, improving the accuracy of the PNLF distance to NGC 300., 20 pages, 11 figures. Accepted for publication in A&A

We report initial results from an ongoing spectroscopic survey of central stars of faint planetary nebulae (PNe), obtained with the Low-Resolution Spectrograph on the Hobby-Eberly Telescope. The six PN nuclei (PNNi) discussed here all have strong emission at the O VI 3811-3834 A doublet, indicative of very high temperatures. Five of them--the nuclei of Ou 2, Kn 61, Kn 15, Abell 72, and Kn 130--belong to the hydrogen-deficient PG 1159 class, showing a strong absorption feature of He II and C IV at 4650-4690 A. Based on exploratory comparisons with synthetic model-atmosphere spectra, and the presence of Ne VIII emission lines, we estimate them to have effective temperatures of order 170,000 K. The central star of Kn 15 has a Wolf-Rayet-like spectrum, with strong and broad emission lines of He II, C IV, N V, and O V-VI. We classify it [WO2], but we note that the N V 4604-4620 A emission doublet is extremely strong, indicating a relatively high nitrogen abundance. Several of the emission lines in Kn 15 vary in equivalent width by factors as large as 1.5 among our four observations from 2019 to 2022, implying significant variations in the stellar mass-loss rate. We encourage spectroscopic monitoring. Follow-up high-time-resolution photometry of these stars would be of interest, given the large fraction of pulsating variables seen among PG 1159 and [WO] PNNi., Comment: Accepted by MNRAS

Assigning names to β-lactamase variants has been inconsistent and has led to confusion in the published literature. The common availability of whole genome sequencing has resulted in an exponential growth in the number of new β-lactamase genes. In November 2021 an international group of β-lactamase experts met virtually to develop a consensus for the way naturally-occurring β-lactamase genes should be named. This document formalizes the process for naming novel β-lactamases, followed by their subsequent publication.

Eight planetary nebulae have been identified as `born-again', a class of object typified by knotty secondary ejecta having low masses ($\sim$$10^{-4}$ M$_{\odot}$) with nearly no hydrogen. Abell 30, the archetype of the class, also belongs to a small subset of planetary nebulae that exhibit extreme abundance discrepancy factors (where Abell 30 is the most extreme), a phenomenon strongly linked to binary star interactions. We report the presence of light curve brightness variations having a period of 1.060 days that are highly suggestive of a binary central star in Abell 30. If confirmed, this detection supports the proposed link between binary central stars and extreme abundance discrepancies., Comment: 5 pages, 3 figures, accepted for publication in MNRAS Letters

A long-term (1995–2016) survey for novae in the nearby Andromeda galaxy (M31) was conducted as part of the Research-Based Science Education initiative. During the course of the survey 180 nights of observation were completed at Kitt Peak, Arizona. A total of 262 novae were either discovered or confirmed, 40 of which have not been previously reported. Of these, 203 novae form a spatially complete sample detected by the KPNO/WIYN 0.9 m telescope within a 20 ′ × 20 ′ field centered on the nucleus of M31. An additional 50 novae are part of a spatially complete sample detected by the KPNO 4 m telescope within a larger 36 ′ × 36 ′ field. Consistent with previous studies, it is found that the spatial distribution of novae in both surveys follows the bulge light of M31 somewhat more closely than the overall background light of the galaxy. After correcting for the limiting magnitude and the spatial and temporal coverage of the surveys, a final nova rate in M31 is found to be R = 40 − 4 + 5 yr−1, which is considerably lower than recent estimates. When normalized to the K-band luminosity of M31, this value yields a luminosity-specific nova rate, ν K = 3.3 ± 0.4 yr − 1 [ 10 10 L ⊙ , K ] − 1 . By scaling the M31 nova rate using the relative infrared luminosities of M31 and our Galaxy, a nova rate of R G = 28 − 4 + 5 yr−1 is found for the Milky Way.

The worldwide distribution of qnr genes found on plasmids and their presence on the chromosomes of aquatic bacteria, such as Vibrio vulnificus, one of the suspected sources, suggests an origin before the development of synthetic quinolones. However, their native function remains unknown. Previous work indicated that expression of qnrVv in V. vulnificus was induced by cold shock. To investigate its role further, we constructed single in-frame deletion mutants in qnrVv and cspA (the gene for cold shock protein) and a double mutant in qnrVv and cspA in V. vulnificus ATCC 17562 to evaluate the response to different environmental conditions and stresses and to exposure to various DNA-damaging agents. We found that qnrVv is involved in resistance to ciprofloxacin, levofloxacin, and mitomycin C and in the cold shock response in V. vulnificus. Moreover, ΔqnrVv and ΔcspA mutants showed slower growth when they were treated with bile salts at 37°C and then shifted to 15°C (cold shock) without bile salts in the medium, with the effect being stronger in the double mutant. This transition may mimic what happens when V. vulnificus is ingested into the gastrointestinal tract and released in its natural environment. Cold shock and bile salts induced the expression of cspA and DNA gyrase and topoisomerase IV genes. However, no induction was found in the ΔqnrVv mutant, suggesting that the qnrVv gene is involved in the response to DNA damage and nucleic acid secondary structure.

The Qnr pentapeptide repeat proteins interact with DNA gyrase and protect it from quinolone inhibition. The two external loops, particularly the larger loop B, of Qnr proteins are essential for quinolone protection of DNA gyrase. The specific QnrB1 interaction sites on DNA gyrase are not known. In this study, we investigated the interaction between GyrA and QnrB1 using site-specific photo-cross-linking of QnrB1 loop B combined with mass spectrometry. We found that amino acid residues 286 to 298 on the tower domain of GyrA interact with QnrB1 and play a key role in QnrB1 protection of gyrase from quinolone inhibition. Alanine replacement of arginine at residue 293 and a small deletion of amino acids 286 to 289 of GyrA resulted in a decrease in the QnrB1-mediated increase in quinolone MICs and also abolished the QnrB1 protection of purified DNA gyrase from ciprofloxacin inhibition.

qnr genes are found in aquatic bacteria and preceded the development of synthetic quinolones. Their natural functions are unknown. We evaluated the expression of chromosomal qnr in Vibrio species in response to environmental stresses and DNA damaging agents. Sub-inhibitory concentrations of quinolones, but not other DNA damaging agents, induced the expression of chromosomal qnr by more than five times in Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio mytili Cold shock also induced the expression of qnr in V. parahaemolyticus, V. vulnificus, and V. mytili, as well as qnrS1 in Escherichia coli qnrS1 induction by cold shock was not altered in ΔihfA or ΔihfB mutants or in a strain over-expressing dnaA, that otherwise directly modulate qnrS1 induction by ciprofloxacin. In contrast, qnrS1 induction by cold shock was reduced in a ΔcspA mutant in the cold shock regulon compared to the wild type. In conclusion, cold shock as well as quinolones induce chromosomal qnr in Vibrio species, and the related qnrS1 in E. coli.

Unlike for classes A and B, a standardized amino acid numbering scheme has not been proposed for the class C (AmpC) β-lactamases, which complicates communication in the field. Here, we propose a scheme developed through a collaborative approach that considers both sequence and structure, preserves traditional numbering of catalytically important residues (Ser64, Lys67, Tyr150, and Lys315), is adaptable to new variants or enzymes yet to be discovered and includes a variation for genetic and epidemiological applications. Fil: Mack, Andrew R.. Case Western Reserve University; Estados Unidos Fil: Barnes, Melissa D.. Case Western Reserve University; Estados Unidos Fil: Taracila, Magdalena A.. Case Western Reserve University; Estados Unidos Fil: Hujer, Andrea M.. Case Western Reserve University; Estados Unidos Fil: Hujer, Kristine M.. Case Western Reserve University; Estados Unidos Fil: Cabot, Gabriel. Instituto de Salud Carlos III; España. Hospital Universitario Son Espases; España Fil: Feldgarden, Michael. National Library Of Medicine; Estados Unidos Fil: Haft, Daniel H.. National Library Of Medicine; Estados Unidos Fil: Klimke, William. National Library Of Medicine; Estados Unidos Fil: Van Den Akker, Focco. Case Western Reserve University; Estados Unidos Fil: Vila, Alejandro Jose. Cleveland VAMC Center for Antimicrobial Resistance and Epidemiology; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina. Plataforma Argentina de Biología Estructural y Metabolómica; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina Fil: Smania, Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina Fil: Haider, Shozeb. Colegio Universitario de Londres; Reino Unido Fil: Papp Wallace, Krisztina M.. Case Western Reserve University; Estados Unidos Fil: Bradford, Patricia A.. No especifíca; Fil: Rossolini, Gian Maria. Florence Careggi University Hospital; Italia. Università degli Studi di Firenze; Italia Fil: Docquier, Jean Denis. Università degli Studi di Siena; Italia Fil: Frère, Jean Marie. Université de Liège; Bélgica Fil: Galleni, Moreno. Université de Liège; Bélgica Fil: Hanson, Nancy D.. Creighton University School of Medicine; Estados Unidos Fil: Oliver, Antonio. Instituto de Salud Carlos III; España. Hospital Universitario Son Espases; España Fil: Plésiat, Patrick. Universite de Bourgogne; Francia. Centre Hospitalier Régional Universitaire; Francia. Centre National de Référence de la Résistance aux Antibiotiques; Francia. Centre National de la Recherche Scientifique; Francia Fil: Poirel, Laurent. University of Fribourg; Suiza Fil: Nordmann, Patrice. University of Fribourg; Suiza Fil: Palzkill, Timothy G.. Baylor College of Medicine; Estados Unidos Fil: Jacoby, George A.. Lahey Hospital and Medical Cente; Estados Unidos Fil: Bush, Karen. Indiana University; Estados Unidos Fil: Bonomo, Robert A.. Case Western Reserve University; Estados Unidos

In a previous study, mutants with enhanced ciprofloxacin resistance (Cip r ) were selected from Escherichia coli J53/pMG252 carrying qnrA1 . Strain J53 Cip r 8-2 showed an increase in the copy number and transcription level of qnrA1 . We sequenced the plasmids on Illumina and MinION platforms.

In a previous study, mutants with enhanced ciprofloxacin resistance (Cip(r)) were selected from Escherichia coli J53/pMG252 carrying qnrA1. Strain J53 Cip(r) 8-2 showed an increase in the copy number and transcription level of qnrA1. We sequenced the plasmids on Illumina and MinION platforms. Parental plasmid pMG252 and plasmid pMG252A from strain J53 Cip(r) 8-2 were almost identical, except for the region containing qnrA1 that in pMG252A contained 4 additional copies of the qnrA1-qacEΔ1-sul1-ISCR1 region.

Introduction: The importance of plasmid-mediated quinolone resistance (PMQR) in Enterobacterales and its high incidence has been emphasised many times. However, a clinical strain carrying more than two PMQR genes is rare. This study sequenced plasmid transconjugants from a donor strain carrying four different PMQR genes to establish their genetic locations. Methods: An Escherichia coli clinical strain displayed remarkable quinolone resistance with a ciprofloxacin MIC of 1024 mg/L carrying four PMQR genes: qnrA1, qepA1, aac(6′)1b-cr and oqxAB. When outcrossed to Escherichia coli J53 AziR, different PMQR genes were transferred and the resulting strains 7C and 8C were chosen for further characterisation. Plasmids were extracted and sequenced by the Illumina and Oxford Nanopore Technologies platforms. S1 nuclease-PFGE was used to estimate the number and size of plasmids. Results: The parental strain had three plasmid bands, as determined by PFGE. Transconjugant 8C obtained three plasmids: pMG336 (162 647 bp, F18:A-:B1:C4) carrying oqxAB; pMG335 carrying qepA1 (73 874 bp, F2:A-:B-); and pMG334 (59 724 bp, IncN (ST5)) with qnrA1 and aac(6′)1b-cr. Interestingly, strain 7C obtained plasmid pMG333 (134 435 bp), which was not present in the parental strain but was an IncN-IncF cointegrate of plasmids pMG334 and pMG335 linked via insertion sequence IS26. Conclusion: This study described the complete nucleotide sequence of three plasmids carrying four PMQR genes in a single strain and the plasmid profile obtained after outcrosses. In addition, it described a cointegrate of two plasmids formed with flanking insertion sequences.

Expression of the quinolone resistance gene qnrS1 is increased by quinolones, but unlike induction of some other qnr genes, the bacterial SOS system is not involved and no lexA box is found upstream. Nonetheless, at least 205 bp of upstream sequence is required for induction to take place. An upstream sequence bound to beads trapped potential binding proteins from cell extracts that were identified by mass spectrometry as Dps, Fis, Ihf, Lrp, CysB, and YjhU. To further elucidate their role, a reporter plasmid linking the qnrS1 upstream sequence to lacZ was introduced into cells of the Keio collection with single-gene deletions and screened for lacZ expression. Mutants in ihfA and ihfB had decreased lacZ induction, while induction in a cysB mutant was increased and dps, fis, lrp, yjhU, and other mutants showed no change. The essential upstream sequence contains potential binding sites for Ihf and DnaA. A dnaA deletion could not be tested because it provides essential functions in cell replication; however, increased dnaA expression decreased qnrS1 induction while decreased dnaA expression enhanced it, implying a role for DnaA as a repressor. In a mobility shift assay, purified IhfA, IhfB, and DnaA proteins (but not CysB) were shown to bind to the upstream segment. Induction decreased in a gyrA quinolone-resistant mutant, indicating that GyrA also has a role. Thus, quinolones acting through proteins DnaA, GyrA, IhfA, and IhfB regulate expression of qnrS1.

Expression of the quinolone resistance gene qnrS1 is increased by quinolones, but unlike induction of some other qnr genes, the bacterial SOS system is not involved and no lexA box is found upstream. Nonetheless, at least 205 bp of upstream sequence is required for induction to take place. An upstream sequence bound to beads trapped potential binding proteins from cell extracts that were identified by mass spectrometry as Dps, Fis, Ihf, Lrp, CysB, and YjhU. To further elucidate their role, a reporter plasmid linking the qnrS1 upstream sequence to lacZ was introduced into cells of the Keio collection with single-gene deletions and screened for lacZ expression. Mutants in ihfA and ihfB had decreased lacZ induction, while induction in a cysB mutant was increased and dps, fis, lrp, yjhU, and other mutants showed no change. The essential upstream sequence contains potential binding sites for Ihf and DnaA. A dnaA deletion could not be tested because it provides essential functions in cell replication; however, increased dnaA expression decreased qnrS1 induction while decreased dnaA expression enhanced it, implying a role for DnaA as a repressor. In a mobility shift assay, purified IhfA, IhfB, and DnaA proteins (but not CysB) were shown to bind to the upstream segment. Induction decreased in a gyrA quinolone-resistant mutant, indicating that GyrA also has a role. Thus, quinolones acting through proteins DnaA, GyrA, IhfA, and IhfB regulate expression of qnrS1.

Bacteria have developed transmissible resistance to almost every antibiotic in clinical use, both synthetic compounds and natural products. The mechanisms are well understood: antibiotic inactivation, target alteration, and drug exclusion. Some mechanisms are antibiotic specific, others class specific, and a few provide resistance to several classes of agents. Resistance genes are ancient and ubiquitous. Sources include antibiotic producers, soil cohabitants that needed protection in order to compete, and housekeeping genes that can be adapted to new uses. Transmission involves conjugative and nonconjugative plasmids, integrons, transposons, insertion sequences, and bacteriophage. Resistance enzymes can be inhibited, and antibiotics can be modified to restore activity, but bacterial enzymes can evolve to lose inhibitor sensitivity, and new mechanisms can be recruited to resist modified drugs. Resistance to available agents is increasing. New antimicrobial agents and fresh ideas are needed to overcome the declining power of our antibacterial weapons.

The planetary nebula luminosity function (PNLF) has been used as an extragalactic distance indicator since the 1980's, but there are still unsolved problems associated with its use. One of the most serious involves PNLF distances beyond ~ 10 Mpc, which tend to be slightly smaller than those of other methods. We consider the implications of previous spectroscopic investigations that found that several of the brightest planetary nebula (PN) candidates in M74 are actually compact supernova remnants (SNRs). Using narrow-band imaging data from the KPNO 4-m telescope, we measure the [O III] $\lambda$5007 and H$\alpha$ fluxes of all the known SNRs in M31 and M33, and test whether those objects could be misidentified as bright PNe at distances beyond ~ 10 Mpc. Our results suggest that compact SNRs are not an important source of contamination in photometric surveys for extragalactic PNe., Comment: 2 pages, 1 figure

We measure the Balmer decrements of 23 of the brightest planetary nebulae (PNe) in the inner bulge ($r \lesssim 3$ arcmin) of M31 and de-redden the bright end of the region's [O III] $\lambda 5007$ planetary nebula luminosity function. We show that the most luminous PNe produce $\gtrsim 1{,}200 \, \rm{L}_{\odot}$ of power in their [O III] $\lambda 5007$ line, implying central star luminosities of at least $\sim 11{,}000 \, \rm{L}_{\odot}$. Even with the most recent accelerated-evolution post-AGB models, such luminosities require central star masses in excess of $0.66 \, \rm{M}_{\odot}$, and main sequence progenitors of at least $\sim 2.5 \, \rm{M}_{\odot}$. Since M31's bulge has very few intermediate-age stars, we conclude that conventional single-star evolution cannot be responsible for these extremely luminous objects. We also present the circumstellar extinctions for the region's bright PNe and demonstrate that the distribution is similar to that found for PNe in the Large Magellanic Cloud, with a median value of $A_{5007} = 0.71$. Finally, we compare our results to extinction measurements made for PNe in the E6 elliptical NGC 4697 and the interacting lenticular NGC 5128. We show that such extinctions are not unusual, and that the existence of very high-mass PN central stars is a general feature of old stellar populations. Our results suggest that single-star population synthesis models significantly underestimate the maximum luminosities and total integrated light of AGB stars., Comment: 13 pages, 14 figures

Qnr is a plasmid-encoded and chromosomally determined protein that protects DNA gyrase and topoisomerase IV from inhibition by quinolones. Despite its prevalence worldwide and existence prior to the discovery of quinolones, its native function is not known. Other synthetic compounds and natural products also target bacterial topoisomerases. A number were studied as molecular probes to gain insight into how Qnr acts. Qnr blocked inhibition by synthetic compounds with somewhat quinolone-like structure that target the GyrA subunit, such as the 2-pyridone ABT-719, the quinazoline-2,4-dione PD 0305970, and the spiropyrimidinetrione pyrazinyl-alkynyl-tetrahydroquinoline (PAT), indicating that Qnr is not strictly quinolone specific, but Qnr did not protect against GyrA-targeting simocyclinone D8 despite evidence that both simocyclinone D8 and Qnr affect DNA binding to gyrase. Qnr did not affect the activity of tricyclic pyrimidoindole or pyrazolopyridones, synthetic inhibitors of the GyrB subunit, or nonsynthetic GyrB inhibitors, such as coumermycin A1, novobiocin, gyramide A, or microcin B17.Thus, in this set of compounds the protective activity of Qnr was confined to those that, like quinolones, trap gyrase on DNA in cleaved complexes.

Quinolone antimicrobials are synthetic and widely used in clinical medicine. Resistance emerged with clinical use and became common in some bacterial pathogens. Mechanisms of resistance include two categories of mutation and acquisition of resistance-conferring genes. Resistance mutations in one or both of the two drug target enzymes, DNA gyrase and DNA topoisomerase IV, are commonly in a localized domain of the GyrA and ParE subunits of the respective enzymes and reduce drug binding to the enzyme-DNA complex. Other resistance mutations occur in regulatory genes that control the expression of native efflux pumps localized in the bacterial membrane(s). These pumps have broad substrate profiles that include quinolones as well as other antimicrobials, disinfectants, and dyes. Mutations of both types can accumulate with selection pressure and produce highly resistant strains. Resistance genes acquired on plasmids can confer low-level resistance that promotes the selection of mutational high-level resistance. Plasmid-encoded resistance is due to Qnr proteins that protect the target enzymes from quinolone action, one mutant aminoglycoside-modifying enzyme that also modifies certain quinolones, and mobile efflux pumps. Plasmids with these mechanisms often encode additional antimicrobial resistances and can transfer multidrug resistance that includes quinolones. Thus, the bacterial quinolone resistance armamentarium is large.

There is no conclusive explanation of why ∼80 per cent of planetary nebulae (PNe) are non-spherical. In the Binary Hypothesis, a binary interaction is a preferred channel to form a non-spherical PN. A fundamental step to corroborate or disprove the Binary Hypothesis is to estimate the binary fraction of central stars of PNe (CSPNe) and compare it with a prediction based on the binary fraction of the progenitor, main-sequence population. In this paper, the second in a series, we search for spatially unresolved I- and J-band flux excess in an extended sample of 34 CSPN by a refined measurement technique with a better quantification of the uncertainties. The detection rate of I- (J-)band flux excess is 32 ± 16 per cent (50 ± 24 per cent). This result is very close to what was obtained in Paper I with a smaller sample. We account conservatively for unobserved cool companions down to brown dwarf luminosities, increasing these fractions to 40 ± 20 per cent (62 ± 30 per cent). This step is very sensitive to the adopted brightness limit of our survey. Accounting for visual companions increases the binary fraction to 46 ± 23 per cent (71 ± 34 per cent). These figures are lower than in Paper I. The error bars are better quantified, but still unacceptably large. Taken at face value, the current CSPN binary fraction is in line with the main-sequence progenitor population binary fraction. However, including white dwarfs companions could increase this fraction by as much as 13 (21) per cent points.

Plasmid-mediated quinolone resistance (PMQR) involves genes for proteins that protect the quinolone targets, an enzyme that inactivates certain quinolones as well as aminoglycosides, and pumps that efflux quinolones. Quinolone susceptibility is reduced by these mechanisms but not to the level of clinical resistance unless chromosomal mutations are also present. PCR primers and conditions for PMQR gene detection are described as well as how to establish a plasmid location.

Plasmid-mediated quinolone resistance (PMQR) involves genes for proteins that protect the quinolone targets, an enzyme that inactivates certain quinolones as well as aminoglycosides, and pumps that efflux quinolones. Quinolone susceptibility is reduced by these mechanisms but not to the level of clinical resistance unless chromosomal mutations are also present. PCR primers and conditions for PMQR gene detection are described as well as how to establish a plasmid location.

We examined 13 qnr-positive and 14 qnr-negative clinical isolates of Escherichia coli for mutations previously seen in a qnr-containing laboratory strain exposed to supra minimum inhibitory concentrations (MICs) of ciprofloxacin. Among the qnr-positive strains, those with ciprofloxacin MICs of ≥ 2 µg/mL had at least one mutation in gyrA. Mutations in parC were present in strains with a ciprofloxacin MIC of ≥ 128 µg/mL. The 6 most ciprofloxacin-resistant strains contained additional plasmid-mediated quinolone resistance determinants. aac(6')-Ib-cr was found in 5 of the 6 strains. Eleven of the 13 strains had alterations in MarR, 9 in SoxR, and 5 had mutations in AcrR. All had elevated expression of at least one efflux pump gene, predominantly acrA (92% of the strains), followed by mdtE (54%) and ydhE (46%). Nine had functionally silent alterations in rfa, two had mutations in gmhB, and one of these also had a mutation in surA. An E. coli with ciprofloxacin MIC of 1024 µg/mL contained 4 different plasmid-mediated quinolone resistance determinants as well as gyrA, parC, parE and pump overexpression mutations. Nine of the 14 qnr-negative strains had mutations in topoisomerase genes with a ciprofloxacin MIC of 0.25 to 256 µg/mL. The three most resistant strains also had mutations in parE. Twelve had alterations in MarR, 10 in SoxR and 5 in AcrR. Ten of the 14 strains had elevated expression of efflux pumps with acrA (71.4%), followed by ydhE (50%) and mdtE (14.3%). A diversity of resistance mechanisms occurs in clinical isolates with and without qnr genes.

Ophthalmia neonatorum, also called neonatal conjunctivitis, acquired during delivery can occur in the first 28 days of life. Commonly caused by the bacterial pathogen Neisseria gonorrhoeae, infection can lead to corneal scarring, perforation of the eye, and blindness. One approach that can be taken to prevent the disease is the use of an ophthalmic prophylaxis, which kills the bacteria on the surface of the eye shortly after birth. Current prophylaxes are based on antibiotic ointments. However, N. gonorrhoeae is resistant to many antibiotics and alternative treatments must be developed before the condition becomes untreatable. This study focused on developing a fatty acid-based prophylaxis. For this, 37 fatty acids or fatty acid derivatives were screened in vitro for fast antigonococcal activity. Seven candidates were identified as bactericidal at 1 mM. These seven were subjected to irritation testing using three separate methods: the bovine corneal opacity and permeability (BCOP) test; the hen’s egg test—chorioallantoic membrane (HET-CAM); and the red blood cell (RBC) lysis assay. The candidates were also tested in artificial tear fluid to determine whether they were effective in this environment. Four of the candidates remained effective. Among these, two lead candidates, monocaprin and myristoleic acid, displayed the best potential as active compounds in the development of a fatty acid-based prophylaxis for prevention of ophthalmia neonatorum., IMPORTANCE Ophthalmia neonatorum is a painful bacterial infection of the eye in newborns. A common cause is Neisseria gonorrhoeae, which has the potential to cause severe damage to the eye and blindness. This study was designed to identify new and unconventional drug candidates that could be used in an eye ointment to prevent the development of this disease. The drugs that were tested were fatty acid-based compounds. The potential candidates were screened for their ability to kill bacteria in tear fluid without causing damage to the eye. This study identified seven candidates that are fast acting and nonirritating, with two strong candidates for potential use in the eye. Further development and testing are now required to formulate an eye treatment incorporating these drugs to prevent ophthalmia neonatorum.

The globular cluster (GC) system of our Galaxy contains four planetary nebulae (PNe): K 648 (or Ps 1) in M15, IRAS 18333-2357 in M22, JaFu 1 in Pal 6, and JaFu 2 in NGC 6441. Because single-star evolution at the low stellar mass of present-epoch GCs was considered incapable of producing visible PNe, their origin presented a puzzle. We imaged the PN JaFu 1 with the Hubble Space Telescope (HST) to obtain photometry of its central star (CS) and high-resolution morphological information. We imaged IRAS 18333-2357 with better depth and resolution, and we analyzed its archival HST spectra to constrain its CS temperature and luminosity. All PNe in Galactic GCs now have high-quality HST data, allowing us to improve CS mass estimates. We find reasonably consistent masses between 0.53 and 0.58 Msun for all four objects, though estimates vary when adopting different stellar evolutionary calculations. The CS mass of IRAS 18333-2357, though, depends strongly on its temperature, which remains elusive due to reddening uncertainties. For all four objects, we consider their CS and nebular masses, their morphologies, and other incongruities to assess the likelihood that these objects formed from binary stars. Although generally limited by uncertainties (~0.02 Msun) in post-AGB tracks and core mass vs. luminosity relations, the high-mass CS in K 648 indicates a binary origin. The CS of JaFu 1 exhibits compact bright [O III] and Halpha emission, like EGB 6, suggesting a binary companion or disk. Evidence is weaker for a binary origin of JaFu 2., Comment: Accepted by Astrophysical Journal

Instead of eliminating infectious diseases, as some had predicted, antibiotic use has inevitably led to the emergence of more antibiotic-resistant pathogens. This chapter reviews the history of our understanding of the processes by which resistance arises. Knowledge of the chemistry and genetics of this phenomenon has allowed the development of improved antibiotics and has made major contributions to molecular biology and the biotechnical revolution.

Plasmid-mediated quinolone resistance (PMQR) caused by qnr genes has been known for 15 years. Information about global distribution and prevalence of qnr genes is abundant, but clinical information concerning infections produced by these isolates and risk factors for their acquisition is limited.Klebsiella pneumoniae blood isolates (n = 227) from a 1 year prospective cohort of patients in Taiwan were studied. MICs of quinolones were determined for all isolates, and multiplex PCR for the presence of PMQR genes and DNA gyrase mutations was applied to all 24 isolates with ciprofloxacin MICs ≥ 0.12 mg/L and a control group of 72 isolates with MICs ≤ 0.06 mg/L.All qnr isolates were in the group with ciprofloxacin MICs ≥ 0.12 mg/L, constituting 9.4% of tested isolates and 3.9% (qnrB 2.6% and qnrS 1.3%) of total isolates. aac(6')-Ib-cr and qepA were not found. Risk factors for qnr included nosocomial infection, bedridden status, surgery within 3 months, non-K1/K2 serotypes and prior antimicrobial use. Ciprofloxacin MIC ≥ 0.12 mg/L was associated with prior quinolone use; in contrast, prior cephalosporin use was more closely linked to the presence of qnr. Fourteen-day mortality was similar in patients infected with qnr-positive versus qnr-negative isolates, but there was a trend for increased in-hospital mortality in patients infected with qnr-positive isolates.In K. pneumoniae blood isolates collected at a hospital in Taiwan, the overall prevalence of qnr genes was 3.9%. Prior quinolone use was linked to increased ciprofloxacin MIC, but not with the prevalence of qnr, which was most strongly linked to exposure to other antimicrobials, especially cephalosporins.

Quinolone antimicrobials are widely used in clinical medicine and are the only current class of agents that directly inhibit bacterial DNA synthesis. Quinolones dually target DNA gyrase and topoisomerase IV binding to specific domains and conformations so as to block DNA strand passage catalysis and stabilize DNA-enzyme complexes that block the DNA replication apparatus and generate double breaks in DNA that underlie their bactericidal activity. Resistance has emerged with clinical use of these agents and is common in some bacterial pathogens. Mechanisms of resistance include mutational alterations in drug target affinity and efflux pump expression and acquisition of resistance-conferring genes. Resistance mutations in one or both of the two drug target enzymes are commonly in a localized domain of the GyrA and ParC subunits of gyrase and topoisomerase IV, respectively, and reduce drug binding to the enzyme-DNA complex. Other resistance mutations occur in regulatory genes that control the expression of native efflux pumps localized in the bacterial membrane(s). These pumps have broad substrate profiles that include other antimicrobials as well as quinolones. Mutations of both types can accumulate with selection pressure and produce highly resistant strains. Resistance genes acquired on plasmids confer low-level resistance that promotes the selection of mutational high-level resistance. Plasmid-encoded resistance is because of Qnr proteins that protect the target enzymes from quinolone action, a mutant aminoglycoside-modifying enzyme that also modifies certain quinolones, and mobile efflux pumps. Plasmids with these mechanisms often encode additional antimicrobial resistances and can transfer multidrug resistance that includes quinolones.

Instrument development for the 24m Giant Magellan Telescope (GMT) is described: current activities, progress, status, and schedule. One instrument team has completed its preliminary design and is currently beginning its final design (GCLEF, an optical 350-950 nm, high-resolution and precision radial velocity echelle spectrograph). A second instrument team is in its conceptual design phase (GMACS, an optical 350-950 nm, medium resolution, 6-10 arcmin field, multi-object spectrograph). A third instrument team is midway through its preliminary design phase (GMTIFS, a near-IR YJHK diffraction-limited imager/integral-field-spectrograph), focused on risk reduction prototyping and design optimization. A fourth instrument team is currently fabricating the 5 silicon immersion gratings needed to begin its preliminary design phase (GMTNIRS, a simultaneous JHKLM high-resolution, AO-fed, echelle spectrograph). And, another instrument team is focusing on technical development and prototyping (MANIFEST, a facility robotic, multifiber feed, with a 20 arcmin field of view). In addition, a medium-field (6 arcmin, 0.06 arcsec/pix) optical imager will support telescope and AO commissioning activities, and will excel at narrow-band imaging. In the spirit of advancing synergies with other groups, the challenges of running an ELT instrument program and opportunities for cross-ELT collaborations are discussed.

The recent availability of large format near-infrared detectors with sub-election readout noise is revolutionizing our approach to wavefront sensing for adaptive optics. However, as with all near-infrared detector technologies, challenges exist in moving from the comfort of the laboratory test-bench into the harsh reality of the observatory environment. As part of the broader adaptive optics program for the GMT, we are developing a near-infrared Lucky Imaging camera for operational deployment at the ANU 2.3 m telescope at Siding Spring Observatory. The system provides an ideal test-bed for the rapidly evolving Selex/SAPHIRA eAPD technology while providing scientific imaging at angular resolution rivalling the Hubble Space Telescope at wavelengths λ = 1.3-2.5 μm.

The Giant Magellan Telescope (GMT) is a 25.4-m diameter, optical/infrared telescope that is being built by an international consortium of universities and research institutions as one of the next generation of Extremely Large Telescopes. The primary mirror of GMT consists of seven 8.4 m borosilicate honeycomb mirror segments that are optically conjugate to seven corresponding segments in the Gregorian secondary mirror. Fabrication is complete for one primary mirror segment and is underway for the next two. The final focal ratio of the telescope is f/8.2, so that the focal plane has an image scale of 1.02 arcsec/mm. GMT will be commissioned using a fast-steering secondary mirror assembly comprised of conventional, rigid segments to provide seeing-limited observations. A secondary mirror with fully adaptive segments will be used in standard operation to additionally enable ground-layer and diffraction-limited adaptive optics. In the seeing limited mode, GMT will provide a 10 arcmin field of view without field correction. A 20 arcmin field of view will be obtained using a wide-field corrector and atmospheric dispersion compensator. The project has recently completed a series of sub-system and system-level preliminary design reviews and is currently preparing to move into the construction phase. This paper summarizes the technical development of the GMT sub-systems and the current status of the GMT project.

Six planetary nebulae (PN) are known in the Kepler space telescope field of view, three newly identified. Of the 5 central stars of PN with useful Kepler data, one, J193110888+4324577, is a short-period, post common envelope binary exhibiting relativistic beaming effects. A second, the central star of the newly identified PN Pa5, has a rare O(He) spectral type and a periodic variability consistent with an evolved companion, where the orbital axis is almost aligned with the line of sight. The third PN, NGC~6826 has a fast rotating central star, something that can only be achieved in a merger. Fourth, the central star of the newly identified PN Kn61, has a PG1159 spectral type and a mysterious semi-periodic light variability which we conjecture to be related to the interplay of binarity with a stellar wind. Finally, the central star of the circular PN A61 does not appear to have a photometric variability above 2 mmag. With the possible exception of the variability of Kn61, all other variability behaviour, whether due to binarity or not, would not easily have been detected from the ground. We conclude, based on very low numbers, that there may be many more close binary or close binary products to be discovered with ultra-high precision photometry. With a larger number of high precision photometric observations we will be able to determine how much higher than the currently known 15 per cent, the short period binary fraction for central stars of PN is likely to be., Comment: 17 pages, 12 figures

Plasmid-carried qnrS1 is derived from Vibrio splendidus chromosomal qnrVS1. qnrVS1 transcripts increased 21- to 34-fold with subinhibitory concentrations of ciprofloxacin but much less with mitomycin. No LexA binding sites were upstream of qnrS1 or qnrVS1 , and similar induction levels were observed in lexA -positive and lexA -negative Escherichia coli strains with native qnrS1 plasmid pMG306 but not with pUC18-cloned qnrS1 or qnrVS1 . Thus, qnrS1 induction by quinolones is independent of the SOS system and requires sequence besides that of the structural gene.

We present [OIII]{\lambda}5007 fluxes and angular diameters for 435 Planetary Nebulae (PN) in the central 10' x 10' region towards the Galactic bulge. Our sample is taken from the new discoveries of the MASH PN surveys as well as previously known PN. This sample accounts for 80% of known PN in this region. Fluxes and diameters are measured from narrow-band imaging with the MOSAIC-II camera on the 4-m Blanco telescope at the Cerro-Tololo Inter-American Observatory. This is the largest (~60 square degrees), uniform [OIII]{\lambda}5007 survey of the inner Galactic bulge ever undertaken. 104 of the objects have measured [OIII]{\lambda}5007, [OIII]{\lambda}4959, H{\alpha} or H{\beta} fluxes from the literature, which we use to undertake a detailed comparison to demonstrate the integrity of our new fluxes. Our independent measurements are in excellent agreement with the very best literature sources over two orders of magnitude, while maintaining good consistency over five orders of magnitude. The excellent resolution and sensitivity of our data allows not only for a robust set of homogenous PN fluxes, but provides greater detail into their intricate, otherwise undetermined [OIII]{\lambda}5007 morphologies. These new, extensive measurements significantly increase the sample of reliable [OIII]{\lambda}5007 fluxes for Galactic bulge PN making it a valuable resource and a prelude to the construction of our new Galactic bulge Planetary Nebula luminosity function (Paper II).

Since glycopeptide-resistant enterococci (GRE) were reported in 1988, they have appeared in hospitals worldwide. Seven van gene cluster types ( vanA , vanB , vanC , vanD , vanE , vanG , and vanL ) are currently known. We investigated a clinical strain of Enterococcus faecium Efm-HS0661 that was isolated in 2006 from an inpatient with intra-abdominal infection in Shanghai. It was resistant to most antimicrobials, including vancomycin (MIC, >256 μg/ml) and teicoplanin (MIC, 96 μg/ml). Glycopeptide resistance could be transferred to E . faecium BM4105RF by conjugation. The donor and its transconjugant were negative by PCR for the known van genes. By cloning and primer walk sequencing, we discovered a novel van gene cluster, designated vanM . The vanM ligase gene was 1,032-bp in length and encoded a 343-amino-acid protein that shared 79.9, 70.8, 66.3, and 78.8% amino acid identity with VanA, VanB, VanD, and VanF, respectively. Although the vanM DNA sequence was closest to vanA , the organization of the vanM gene cluster was most similar to that of vanD . Upstream from the vanM cluster was an IS 1216- like element, which may play a role in the dissemination of this resistance determinant. Liquid chromatography-mass spectrometry analysis of peptidoglycan precursors extracted from the VanM-type strain Efm-HS0661 treated with vancomycin or teicoplanin revealed a modified precursor (UDP- N -acetylmuramic acid [MurNAc]-tetrapeptide- d -Lac), indicating that VanM, like VanA, confers glycopeptide resistance by the inducible synthesis of precursor ending in d -Ala- d -Lac.

Ciprofloxacin was introduced for treatment of patients with cholera in Bangladesh because of resistance to other agents, but its utility has been compromised by the decreasing ciprofloxacin susceptibility of Vibrio cholerae over time. We correlated levels of susceptibility and temporal patterns with the occurrence of mutation in gyrA , which encodes a subunit of DNA gyrase, followed by mutation in parC , which encodes a subunit of DNA topoisomerase IV. We found that ciprofloxacin activity was more recently further compromised in strains containing qnrVC3 , which encodes a pentapeptide repeat protein of the Qnr subfamily, members of which protect topoisomerases from quinolone action. We show that qnrVC3 confers transferable low-level quinolone resistance and is present within a member of the SXT integrating conjugative element family found commonly on the chromosomes of multidrug-resistant strains of V. cholerae and on the chromosomes of Escherichia coli transconjugants constructed in the laboratory. Thus, progressive increases in quinolone resistance in V. cholerae are linked to cumulative mutations in quinolone targets and most recently to a qnr gene on a mobile multidrug resistance element, resulting in further challenges for the antimicrobial therapy of cholera.

Recent Halpha surveys such as SHS and IPHAS have improved the completeness of the Galactic planetary nebula (PN) census. We now know of ~3,000 PNe in the Galaxy, but this is far short of most estimates, typically ~25,000 or more for the total population. The size of the Galactic PN population is required to derive an accurate estimate of the chemical enrichment rates of nitrogen, carbon, and helium. In addition, a high PN count (~20,000) is strong evidence that most 1-8 Msun main sequence stars will go through a PN phase, while a low count (, 11 pages, 7 figures, accepted for publication in the Publications of the Astronomical Society of Australia (PASA), full resolution paper available at http://www.wiyn.org/jacoby_pasa.pdf

In 50 years, the International Conference on Antimicrobial Agents and Chemotherapy—known as ICAAC—has grown from a small, almost entirely U.S.-based meeting to the largest international conference on antimicrobial agents, infectious diseases, and microbiology. One key reason for this success is an enduring focus on scientific excellence and innovation. During those five decades, domestic and international attendance and numbers of scientific presentations has grown steadily, while scientific rigor remained steady.

SUMMARYAlthough plasmid-mediated quinolone resistance (PMQR) was thought not to exist before its discovery in 1998, the past decade has seen an explosion of research characterizing this phenomenon. The best-described form of PMQR is determined by theqnrgroup of genes. These genes, likely originating in aquatic organisms, code for pentapeptide repeat proteins. These proteins reduce susceptibility to quinolones by protecting the complex of DNA and DNA gyrase or topoisomerase IV enzymes from the inhibitory effect of quinolones. Two additional PMQR mechanisms were recently described.aac(6′)-Ib-crencodes a variant aminoglycoside acetyltransferase with two amino acid alterations allowing it to inactivate ciprofloxacin through the acetylation of its piperazinyl substituent.oqxABandqepAencode efflux pumps that extrude quinolones. All of these genes determine relatively small increases in the MICs of quinolones, but these changes are sufficient to facilitate the selection of mutants with higher levels of resistance. The contribution of these genes to the emergence of quinolone resistance is being actively investigated. Several factors suggest their importance in this process, including their increasing ubiquity, their association with other resistance elements, and their emergence simultaneous with the expansion of clinical quinolone resistance. Of concern, these genes are not yet being taken into account in resistance screening by clinical microbiology laboratories.

Recently, several plasmid-mediated quinolone resistance (PMQR) genes conferring low levels of quinolone resistance have been discovered. To evaluate the temporal change in the prevalence of PMQR genes over a decade in a tertiary hospital in the Republic of Korea, we selected every fifth isolate of Escherichia coli and Klebsiella pneumoniae and every third isolate of Enterobacter cloacae between 1998 and 2001 and between 2005 and 2006 from a collection of blood isolates. Six PMQR genes [ qnrA , qnrB , qnrC , qnrS , aac(6′)-Ib-cr , and qepA ] were screened by multiplex PCR and then confirmed by direct sequencing, and the aac(6′)-Ib -positive PCR products were digested with BtsCI to identify the aac(6′)-Ib-cr variant. Of 461 isolates, 37 (8%) had one of the six PMQR genes; 13 (5%) of 261 E. coli strains, 13 (10%) of 135 K. pneumoniae strains, and 11 (17%) of 65 E. cloacae strains. qnrB was the most common PMQR gene and was found as early as 1998, whereas qnrS , aac(6′)-Ib - cr , and qepA emerged after 2000. None of the isolates carried qnrA or qnrC . Ciprofloxacin resistance increased over time ( P < 0.001), and the overall prevalence of PMQR genes tended to increase ( P = 0.20). PMQR-positive isolates had significantly higher ciprofloxacin resistance and multidrug resistance rates ( P = 0.005 and P < 0.001, respectively). The increasing frequency of ciprofloxacin resistance in Enterobacteriaceae was associated with an increasing prevalence of PMQR genes, and this change involved an increase in the diversity of the PMQR genes and also an increase in the prevalence of the mutations in gyrA , parC , or both in PMQR-positive strains but not PMQR-negative strains.

SUMMARYAmpC β-lactamases are clinically important cephalosporinases encoded on the chromosomes of many of the Enterobacteriaceae and a few other organisms, where they mediate resistance to cephalothin, cefazolin, cefoxitin, most penicillins, and β-lactamase inhibitor-β-lactam combinations. In many bacteria, AmpC enzymes are inducible and can be expressed at high levels by mutation. Overexpression confers resistance to broad-spectrum cephalosporins including cefotaxime, ceftazidime, and ceftriaxone and is a problem especially in infections due to Enterobacter aerogenes and Enterobacter cloacae, where an isolate initially susceptible to these agents may become resistant upon therapy. Transmissible plasmids have acquired genes for AmpC enzymes, which consequently can now appear in bacteria lacking or poorly expressing a chromosomal blaAmpCgene, such as Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis. Resistance due to plasmid-mediated AmpC enzymes is less common than extended-spectrum β-lactamase production in most parts of the world but may be both harder to detect and broader in spectrum. AmpC enzymes encoded by both chromosomal and plasmid genes are also evolving to hydrolyze broad-spectrum cephalosporins more efficiently. Techniques to identify AmpC β-lactamase-producing isolates are available but are still evolving and are not yet optimized for the clinical laboratory, which probably now underestimates this resistance mechanism. Carbapenems can usually be used to treat infections due to AmpC-producing bacteria, but carbapenem resistance can arise in some organisms by mutations that reduce influx (outer membrane porin loss) or enhance efflux (efflux pump activation).

Deep wide-field H$\alpha$+[NII] imaging around the Virgo cluster giant elliptical galaxy M86 reveals a highly complex and disturbed ISM/ICM. The most striking feature is a set of H$\alpha$ filaments which clearly connect M86 with the nearby disturbed spiral NGC 4438 (23$'$=120 kpc projected away), providing strong evidence for a previously unrecognized collision between them. Spectroscopy of selected regions show a fairly smooth velocity gradient between M86 and NGC 4438, consistent with the collision scenario. Such a collision would impart significant energy into the ISM of M86, probably heating the gas and acting to prevent the gas from cooling to form stars. We propose that cool gas stripped from NGC 4438 during the collision and deposited in its wake is heated by shocks, ram pressure drag, or thermal conduction, producing most of the H$\alpha$ filaments. Some H$\alpha$ filaments are associated with the well-known ridge of bright X-ray emission to the NW of the nucleus, suggesting that the collision is responsible for peculiarities of M86 previously ascribed to other effects. M86 is radio-quiet, thus AGN heating is unlikely to play a significant role. The M86 system has implications for understanding the role of gravitational interactions in the heating of the ISM in ellipticals, and how collisions in clusters transform galaxies., Comment: 6 pages, 2 figures. For high-resolution images, see http://www.astro.yale.edu/tal/research/index.html