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Article ; Online: In vitro and in vivo activity of GT-1, a novel siderophore cephalosporin, and GT-055, a broad-spectrum β-lactamase inhibitor, against biothreat and ESKAPE pathogens.

Halasohoris, Stephanie A / Scarff, Jennifer M / Pysz, Lisa M / Lembirik, Sanae / Lemmon, Margaret M / Biek, Donald / Hannah, Brendan / Zumbrun, Steven D / Panchal, Rekha G

The Journal of antibiotics

2021 Volume 74, Issue 12, Page(s) 884–892

Abstract: Antimicrobial-resistance (AMR) has become an increasingly difficult issue to overcome for bacteria associated with both community- and hospital-acquired infections as well as potential biodefense threats. The need to identify new therapeutics of novel ... ...

Abstract	Antimicrobial-resistance (AMR) has become an increasingly difficult issue to overcome for bacteria associated with both community- and hospital-acquired infections as well as potential biodefense threats. The need to identify new therapeutics of novel classes and/or with unique mechanisms is critical to combatting AMR in the coming years. GT-1 (LCB10-0200), a siderophore-linked cephalosporin, is one such novel option and is formulated to be used either alone or in combination with a novel broad-spectrum β-lactamase inhibitor, GT-055 (LCB18-055). This study assessed the in vitro and in vivo efficacy of GT-1 and GT-055 against a broad array of multi-drug resistant and biothreat pathogens. Here, we demonstrated sub-4 µg ml
MeSH term(s)	Animals ; Anti-Bacterial Agents/pharmacology ; Anti-Bacterial Agents/therapeutic use ; Biological Warfare Agents ; Cephalosporins/pharmacology ; Cephalosporins/therapeutic use ; Ciprofloxacin/pharmacology ; Drug Resistance, Multiple, Bacterial ; Mice ; Mice, Inbred BALB C ; Microbial Sensitivity Tests ; Plague/drug therapy ; Plague/microbiology ; Siderophores/pharmacology ; Yersinia pestis/drug effects ; beta-Lactamase Inhibitors/pharmacology ; beta-Lactamase Inhibitors/therapeutic use
Chemical Substances	Anti-Bacterial Agents ; Biological Warfare Agents ; Cephalosporins ; GT-1 siderophore cephalosporin ; Siderophores ; beta-Lactamase Inhibitors ; Ciprofloxacin (5E8K9I0O4U)
Language	English
Publishing date	2021-09-14
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	390800-8
ISSN	1881-1469 ; 0021-8820 ; 0368-3532
ISSN (online)	1881-1469
ISSN	0021-8820 ; 0368-3532
DOI	10.1038/s41429-021-00472-9
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Zs.A 207: Show issues

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Article ; Online: Emergence of Lyme Disease on Treeless Islands, Scotland, United Kingdom.

Millins, Caroline / Leo, Walter / MacInnes, Isabell / Ferguson, Johanne / Charlesworth, Graham / Nayar, Donald / Davison, Reece / Yardley, Jonathan / Kilbride, Elizabeth / Huntley, Selene / Gilbert, Lucy / Viana, Mafalda / Johnson, Paul / Biek, Roman

Emerging infectious diseases

2021 Volume 27, Issue 2, Page(s) 538–546

Abstract: Lyme disease is usually associated with forested habitats but has recently emerged on treeless islands in the Western Isles of Scotland. The environmental and human components of Lyme disease risk in open habitats remain unknown. We quantified the ... ...

Abstract	Lyme disease is usually associated with forested habitats but has recently emerged on treeless islands in the Western Isles of Scotland. The environmental and human components of Lyme disease risk in open habitats remain unknown. We quantified the environmental hazard and risk factors for human tick bite exposure among treeless islands with low and high Lyme disease incidence in the Western Isles. We found a higher prevalence of Borrelia burgdorferi sensu lato-infected ticks on high-incidence than on low-incidence islands (6.4% vs. 0.7%); we also found that residents of high-incidence islands reported increased tick bite exposure. Most tick bites (72.7%) occurred <1 km from the home, including many in home gardens. Residents of high Lyme disease incidence islands reported increasing problems with ticks; many suggested changing deer distribution as a potential driver. We highlight the benefits of an integrated approach in understanding the factors that contribute to Lyme disease emergence.
MeSH term(s)	Animals ; Deer ; Humans ; Islands ; Ixodes ; Lyme Disease/epidemiology ; Nymph ; Scotland/epidemiology ; United Kingdom
Language	English
Publishing date	2021-01-26
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1380686-5
ISSN	1080-6059 ; 1080-6040
ISSN (online)	1080-6059
ISSN	1080-6040
DOI	10.3201/eid2702.203862
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Article: Fosfomycin and Comparator Activity Against Select Enterobacteriaceae, Pseudomonas, and Enterococcus Urinary Tract Infection Isolates from the United States in 2012.

Keepers, Tiffany R / Gomez, Marcela / Celeri, Chris / Krause, Kevin M / Biek, Donald / Critchley, Ian

Infectious diseases and therapy

2017 Volume 6, Issue 2, Page(s) 233–243

Abstract: Introduction: Fosfomycin is a broad-spectrum cell wall active agent that inhibits the MurA enzyme involved in peptidoglycan synthesis and is FDA-approved for treatment of uncomplicated urinary tract infections (UTIs) caused by Escherichia coli and ... ...

Abstract	Introduction: Fosfomycin is a broad-spectrum cell wall active agent that inhibits the MurA enzyme involved in peptidoglycan synthesis and is FDA-approved for treatment of uncomplicated urinary tract infections (UTIs) caused by Escherichia coli and Enterococcus faecalis in women. Data regarding the susceptibility of recent UTI isolates to fosfomycin are limited. Methods: This study compared the fosfomycin susceptibility of 658 US UTI isolates with susceptibility to ciprofloxacin, levofloxacin, nitrofurantoin, and trimethoprim/sulfamethoxazole (SXT). Isolates included E. coli (n = 257), Klebsiella spp. (n = 156), Enterobacter spp. (n = 79), Pseudomonas aeruginosa (n = 60), E. faecalis (n = 54), and Proteus spp. (n = 52). Extended-spectrum β-lactamase (ESBL)-producing E. coli, Klebsiella spp., and Proteus mirabilis, ceftazidime-nonsusceptible P. aeruginosa and Enterobacter spp., and vancomycin-nonsusceptible E. faecalis were included. Results: Overall, the minimum concentration inhibiting 50% of isolates (MIC Conclusion: These results demonstrate that fosfomycin has in vitro activity against many US UTI isolates, including drug-resistant isolates, and may provide another therapeutic option for treatment of UTIs caused by antibiotic-resistant pathogens.
Language	English
Publishing date	2017-03-11
Publishing country	New Zealand
Document type	Journal Article
ZDB-ID	2701611-0
ISSN	2193-6382 ; 2193-8229
ISSN (online)	2193-6382
ISSN	2193-8229
DOI	10.1007/s40121-017-0150-5
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Article ; Online: Emergence of Lyme Disease on Treeless Islands, Scotland, United Kingdom

Caroline Millins / Walter Leo / Isabell MacInnes / Johanne Ferguson / Graham Charlesworth / Donald Nayar / Reece Davison / Jonathan Yardley / Elizabeth Kilbride / Selene Huntley / Lucy Gilbert / Mafalda Viana / Paul Johnson / Roman Biek

Emerging Infectious Diseases, Vol 27, Iss 2, Pp 538-

2021 Volume 546

Abstract	Lyme disease is usually associated with forested habitats but has recently emerged on treeless islands in the Western Isles of Scotland. The environmental and human components of Lyme disease risk in open habitats remain unknown. We quantified the environmental hazard and risk factors for human tick bite exposure among treeless islands with low and high Lyme disease incidence in the Western Isles. We found a higher prevalence of Borrelia burgdorferi sensu lato–infected ticks on high-incidence than on low-incidence islands (6.4% vs. 0.7%); we also found that residents of high-incidence islands reported increased tick bite exposure. Most tick bites (72.7%) occurred <1 km from the home, including many in home gardens. Residents of high Lyme disease incidence islands reported increasing problems with ticks; many suggested changing deer distribution as a potential driver. We highlight the benefits of an integrated approach in understanding the factors that contribute to Lyme disease emergence.
Keywords	Lyme disease ; treeless habitats ; Ixodes ricinus ; peridomestic ; tick bite exposure ; deer ; Medicine ; R ; Infectious and parasitic diseases ; RC109-216
Subject code	610
Language	English
Publishing date	2021-02-01T00:00:00Z
Publisher	Centers for Disease Control and Prevention
Document type	Article ; Online
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Article: Effect of In Vitro Testing Parameters on Ceftazidime-Avibactam Minimum Inhibitory Concentrations.

Keepers, Tiffany R / Gomez, Marcela / Biek, Donald / Critchley, Ian / Krause, Kevin M

International scholarly research notices

2015 Volume 2015, Page(s) 489547

Abstract: Effects of varying in vitro susceptibility testing parameters of the broth microdilution assay on ceftazidime-avibactam MICs were determined and compared to meropenem and piperacillin-tazobactam for 9 Enterobacteriaceae and 4 Pseudomonas aeruginosa ... ...

Abstract	Effects of varying in vitro susceptibility testing parameters of the broth microdilution assay on ceftazidime-avibactam MICs were determined and compared to meropenem and piperacillin-tazobactam for 9 Enterobacteriaceae and 4 Pseudomonas aeruginosa isolates. The effect of varying incubation conditions (ambient air or 5% CO2), pH of medium, medium composition (cation-adjusted Mueller Hinton Broth with and without laked horse blood and Haemophilus Test Medium), cation content of the medium, and inoculum density were tested. Most variations had no effect on ceftazidime-avibactam MIC values (no more than a 2-fold change). However, acidic pH or high inoculum resulted in 4- to 16-fold changes in MIC, which was similar to those observed for meropenem and piperacillin-tazobactam under these conditions. Overall, this study shows that slight variations in testing parameters during routine MIC testing will likely have no significant effect on ceftazidime-avibactam MIC values.
Language	English
Publishing date	2015-05-19
Publishing country	United States
Document type	Journal Article
ZDB-ID	2778458-7
ISSN	2356-7872
ISSN	2356-7872
DOI	10.1155/2015/489547
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Article ; Online: Efficacy of ceftaroline fosamil in a staphylococcal murine pneumonia model.

Bhalodi, Amira A / Crandon, Jared L / Biek, Donald / Nicolau, David P

Antimicrobial agents and chemotherapy

2012 Volume 56, Issue 12, Page(s) 6160–6165

Abstract: Ceftaroline fosamil is a cephalosporin with activity against Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus (MRSA). The objective of this study was to characterize the dose-response relationship of ceftaroline fosamil ... ...

Abstract	Ceftaroline fosamil is a cephalosporin with activity against Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus (MRSA). The objective of this study was to characterize the dose-response relationship of ceftaroline fosamil against S. aureus in an immunocompromised murine pneumonia model, as well as to evaluate the efficacy of the humanized regimen of 600 mg intravenously (i.v.) every 12 h. Seventeen S. aureus (2 methicillin-susceptible Staphylococcus aureus [MSSA], 15 MRSA) isolates with ceftaroline MICs of 0.5 to 4 μg/ml were utilized. The pharmacokinetics of ceftaroline in serum and epithelial lining fluid (ELF) were evaluated to determine bronchopulmonary exposure profiles in infected and uninfected animals, using single and human-simulated doses. Serum fT>MIC (the percentage of time that free drug concentrations remain above the MIC) of 17% to 43% was required to produce a 1-log(10) kill in the dose-ranging studies. These targets were readily achieved with the humanized exposure profile, where decreases of 0.64 to 1.95 log(10) CFU were observed against 13 MRSA and both MSSA isolates tested. When taken as a composite, the fT>MICs required for stasis and a 1-log(10) kill were 16% and 41%, respectively. ELF concentrations were similar to serum concentrations across the dosing interval in infected and uninfected animals. The serum fT>MIC targets required in this lung infection model were similar to those observed with ceftaroline against S. aureus in a murine thigh infection model. Exposures simulating the human dose of 600 mg i.v. every 12 h achieved pharmacodynamic targets against MRSA and MSSA considered susceptible by current U.S. FDA breakpoints.
MeSH term(s)	Animals ; Anti-Bacterial Agents/administration & dosage ; Anti-Bacterial Agents/pharmacokinetics ; Anti-Bacterial Agents/therapeutic use ; Cephalosporins/pharmacology ; Colony Count, Microbial ; Dose-Response Relationship, Drug ; Female ; Humans ; Immunocompromised Host ; Lung/microbiology ; Methicillin-Resistant Staphylococcus aureus ; Mice ; Mice, Inbred BALB C ; Microbial Sensitivity Tests ; Neutropenia/complications ; Pneumonia, Staphylococcal/drug therapy ; Pneumonia, Staphylococcal/microbiology ; Protein Binding ; Ceftaroline
Chemical Substances	Anti-Bacterial Agents ; Cephalosporins
Language	English
Publishing date	2012-09-17
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	217602-6
ISSN	1098-6596 ; 0066-4804
ISSN (online)	1098-6596
ISSN	0066-4804
DOI	10.1128/AAC.01078-12
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Zs.A 809: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Ceftaroline fosamil for the treatment of acute bacterial skin and skin structure infections.

Beresford, Eric / Biek, Donald / Jandourek, Alena / Mawal, Yogesh / Riccobene, Todd / Friedland, H David

Expert review of clinical pharmacology

2014 Volume 7, Issue 2, Page(s) 123–135

Abstract: Skin infections have traditionally been classified by the US FDA as uncomplicated and complicated. In August 2010, the FDA released a new guidance document for the development of drugs to treat acute bacterial skin and skin structure infections (ABSSSI) ... ...

Abstract	Skin infections have traditionally been classified by the US FDA as uncomplicated and complicated. In August 2010, the FDA released a new guidance document for the development of drugs to treat acute bacterial skin and skin structure infections (ABSSSI) and this was updated in 2013. Several new issues were addressed and henceforth skin infections in clinical trials were termed ABSSSI. In the USA, the annual prevalence of methicillin-resistant Staphylococcus aureus-related skin infections have continuously increased from 32.7% in 1998 to 53.8% in 2007. Ceftaroline fosamil is the only cephalosporin approved in the USA for monotherapy treatment of ABSSSI including infections caused by methicillin-resistant S. aureus. The efficacy of ceftaroline fosamil was shown in the CANVAS clinical trials. The CANVAS Day-3 analyses met an earlier, primary efficacy time point requested by the FDA. Ceftaroline has minimal drug-drug interactions, is well tolerated and possesses the safety profile associated with the cephalosporin class.
MeSH term(s)	Acute Disease ; Anti-Bacterial Agents/adverse effects ; Anti-Bacterial Agents/therapeutic use ; Cephalosporins/adverse effects ; Cephalosporins/therapeutic use ; Drug Approval ; Drug Design ; Drug Interactions ; Humans ; Methicillin-Resistant Staphylococcus aureus/isolation & purification ; Skin Diseases, Bacterial/drug therapy ; Skin Diseases, Bacterial/microbiology ; Staphylococcal Infections/drug therapy ; Staphylococcal Infections/microbiology ; United States ; United States Food and Drug Administration ; Ceftaroline
Chemical Substances	Anti-Bacterial Agents ; Cephalosporins
Language	English
Publishing date	2014-02-05
Publishing country	England
Document type	Journal Article ; Review
ISSN	1751-2441
ISSN (online)	1751-2441
DOI	10.1586/17512433.2014.884457
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Article ; Online: Ceftaroline fosamil: a novel broad-spectrum cephalosporin with expanded anti-Gram-positive activity.

Biek, Donald / Critchley, Ian A / Riccobene, Todd A / Thye, Dirk A

The Journal of antimicrobial chemotherapy

2010 Volume 65 Suppl 4, Page(s) iv9–16

Abstract: Ceftaroline fosamil is a novel cephalosporin with broad-spectrum activity against Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Streptococcus pneumoniae, and common Gram-negative organisms. ... ...

Abstract	Ceftaroline fosamil is a novel cephalosporin with broad-spectrum activity against Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Streptococcus pneumoniae, and common Gram-negative organisms. The activity of ceftaroline against MRSA is attributed to its ability to bind to penicillin-binding protein (PBP) 2a with high affinity and inhibit the biochemical activity of PBP 2a more efficiently than other presently available β-lactams. The activity of ceftaroline against MRSA and the β-haemolytic streptococci makes it an attractive monotherapy agent for the treatment of complicated skin and skin structure infections (cSSSIs). Recent profiling and surveillance studies have shown that ceftaroline is active against contemporary skin pathogens collected from US and European medical centres in 2008. The mean free drug %T > MIC (percentage of time the drug concentration remains above the MIC) needed for stasis ranged from 26% for S. aureus to 39% for S. pneumoniae in the murine thigh infection model. Pharmacokinetic and pharmacodynamic target attainment predictions for 600 mg of ceftaroline fosamil every 12 h showed that the mean %T > MICs for which plasma free-drug concentrations exceeded an MIC of 1 and 2 mg/L were 71% and 51% of the dosing interval, respectively. For a 40% T > MIC target, the predicted attainments for infections due to pathogens for which ceftaroline MICs were 1 or 2 mg/L were 100% and 90%, respectively. Clinical and microbiological successes of ceftaroline fosamil in treating cSSSIs were demonstrated in two Phase III clinical studies, in which 96.8% of all baseline cSSSI isolates from the microbiologically evaluable population were inhibited by ceftaroline at ≤ 2 mg/L. Ceftaroline fosamil is a promising broad-spectrum agent for the treatment of cSSSIs.
MeSH term(s)	Animals ; Anti-Bacterial Agents/chemistry ; Anti-Bacterial Agents/pharmacokinetics ; Anti-Bacterial Agents/pharmacology ; Anti-Bacterial Agents/therapeutic use ; Cephalosporins/chemistry ; Cephalosporins/pharmacokinetics ; Cephalosporins/pharmacology ; Cephalosporins/therapeutic use ; Clinical Trials, Phase III as Topic ; Drug Resistance, Bacterial ; Gram-Negative Bacteria/drug effects ; Gram-Positive Bacteria/drug effects ; Humans ; Methicillin-Resistant Staphylococcus aureus/drug effects ; Mice ; Microbial Sensitivity Tests ; Skin Diseases, Bacterial/drug therapy ; Skin Diseases, Bacterial/microbiology ; Soft Tissue Infections/drug therapy ; Soft Tissue Infections/microbiology ; Treatment Outcome ; Ceftaroline
Chemical Substances	Anti-Bacterial Agents ; Cephalosporins
Language	English
Publishing date	2010-11-29
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	191709-2
ISSN	1460-2091 ; 0305-7453
ISSN (online)	1460-2091
ISSN	0305-7453
DOI	10.1093/jac/dkq251
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Zs.A 1197: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Evaluation of ceftaroline activity versus ceftriaxone against clinical isolates of Streptococcus pneumoniae with various susceptibilities to cephalosporins in an in vitro pharmacokinetic/pharmacodynamic model.

Steed, Molly E / Vidaillac, Céline / Winterfield, Patricia / Biek, Donald / Rybak, Michael J

Antimicrobial agents and chemotherapy

2012 Volume 56, Issue 5, Page(s) 2691–2695

Abstract: Drug resistance in Streptococcus pneumoniae, a frequent pathogen in community-acquired pneumonia, is increasing. Ceftaroline (active metabolite of ceftaroline fosamil) is a broad-spectrum intravenous cephalosporin with activity in vitro against drug- ... ...

Abstract	Drug resistance in Streptococcus pneumoniae, a frequent pathogen in community-acquired pneumonia, is increasing. Ceftaroline (active metabolite of ceftaroline fosamil) is a broad-spectrum intravenous cephalosporin with activity in vitro against drug-resistant Gram-positive organisms. We investigated ceftaroline at 600 mg every 12 h (q12h) (maximum concentration of the free, unbound drug in serum [fC(max)] is 15.2 μg/ml, and half-life [T(1/2)] is 2.5 h) versus ceftriaxone at 1 g q24h (fC(max) = 23 μg/ml, T(1/2) = 8 h) against six clinical S. pneumoniae isolates in a one-compartment in vitro pharmacokinetic/pharmacodynamic 96-h model (starting inoculum of 10(7) CFU/ml). Differences in CFU/ml (at 24 to 96 h) were evaluated by analysis of variance with a Tukey's post hoc test. Bactericidal activity was defined as a ≥ 3 log(10) CFU/ml decrease from the initial inoculum. Ceftaroline MICs were 0.06, 0.015, ≤ 0.008, 0.25, 0.25, and 0.5 μg/ml, and ceftriaxone MICs were 0.5, 0.25, 0.25, 4, 4, and 8 μg/ml for SP 1477, SP 669, SP 132, SP 211, SP 90, and SP 1466, respectively. Against the ceftaroline- and ceftriaxone-susceptible strain SP 1477, ceftaroline displayed sustained bactericidal activity (3 to 96 h, -5.49 log(10) CFU/ml) and was significantly (P ≤ 0.012) better than ceftriaxone (72 to 96 h, -2.03 log(10) CFU/ml). Against the ceftriaxone-resistant strains, ceftaroline displayed sustained bactericidal activity at 96 h and was significantly better than ceftriaxone (SP211 [-5.91 log(10) CFU/ml, P ≤ 0.002], SP 90 [-5.26 log(10) CFU/ml, P ≤ 0.008], and SP1466 [-5.14 log(10) CFU/ml, P ≤ 0.042]). Ceftaroline was the more effective drug and displayed sustained bactericidal activity. Ceftaroline fosamil may provide a therapeutic option to treat ceftriaxone-resistant S. pneumoniae infections.
MeSH term(s)	Anti-Bacterial Agents/pharmacokinetics ; Anti-Bacterial Agents/pharmacology ; Ceftriaxone/pharmacokinetics ; Ceftriaxone/pharmacology ; Cephalosporin Resistance/drug effects ; Cephalosporins/pharmacokinetics ; Cephalosporins/pharmacology ; Community-Acquired Infections/microbiology ; Culture Media ; Half-Life ; Humans ; Microbial Sensitivity Tests ; Models, Biological ; Pneumonia, Pneumococcal/microbiology ; Streptococcus pneumoniae/drug effects ; Streptococcus pneumoniae/growth & development ; Streptococcus pneumoniae/isolation & purification ; Ceftaroline
Chemical Substances	Anti-Bacterial Agents ; Cephalosporins ; Culture Media ; Ceftriaxone (75J73V1629)
Language	English
Publishing date	2012-02-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	217602-6
ISSN	1098-6596 ; 0066-4804
ISSN (online)	1098-6596
ISSN	0066-4804
DOI	10.1128/AAC.06185-11
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Article: P1 plasmid partition: in vivo evidence for the ParA- and ParB-mediated formation of an anchored parS complex in the absence of a partner parS.

Edgar, Rotem / Biek, Donald / Yarmolinsky, Michael

Molecular microbiology

2006 Volume 59, Issue 1, Page(s) 276–287

Abstract: ParA and ParB proteins and cis-acting site, parS, are required to partition plasmid P1 faithfully to daughter cells. The process may initiate from plasmids paired by ParB at which recruited ParA then acts to effect the separation. We previously reported ... ...

Abstract	ParA and ParB proteins and cis-acting site, parS, are required to partition plasmid P1 faithfully to daughter cells. The process may initiate from plasmids paired by ParB at which recruited ParA then acts to effect the separation. We previously reported evidence for ParB-mediated pairing of parS sites on plasmids in the absence of ParA. In DNA gyrase-inhibited cells, the pairing prevented diffusion of transcription-generated positive supercoils. This supercoil trapping was almost entirely in plasmid dimers, where the location of the parS sites in cis facilitated their pairing. Here we show that the addition of ParA blocked supercoil diffusion also in plasmid monomers. The possibility that this result is attributed to an enhancement by ParA of ParB-mediated pairing in trans is consistent with our finding that ParA appeared to partially suppress the pairing defect of two mutant ParB proteins. However, enhanced pairing alone could not account for the diffusion barrier in plasmid monomers; it was manifest in monomers even when they were largely devoid of partners in the same cell. Apparently, ParA altered the ParB-parS complex such that it could no longer swivel, most likely by anchoring it, a reaction of probable relevance to partition.
MeSH term(s)	Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; DNA Gyrase/metabolism ; DNA, Bacterial/chemistry ; DNA, Bacterial/metabolism ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Gene Expression Regulation, Bacterial ; Nucleic Acid Conformation ; Plasmids/chemistry ; Plasmids/genetics ; Plasmids/metabolism ; Topoisomerase II Inhibitors
Chemical Substances	Bacterial Proteins ; DNA, Bacterial ; Topoisomerase II Inhibitors ; chromosome partition proteins, bacterial ; DNA Gyrase (EC 5.99.1.3)
Language	English
Publishing date	2006-01
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	619315-8
ISSN	1365-2958 ; 0950-382X
ISSN (online)	1365-2958
ISSN	0950-382X
DOI	10.1111/j.1365-2958.2005.04933.x
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