scholarly journals Structural studies of proteins involved in the activity of novel antibiotics

2014 ◽  
Vol 70 (a1) ◽  
pp. C710-C710
Author(s):  
Lucile Moynie ◽  
Luana Ferrara ◽  
Antoni Tortajada ◽  
James Naismith
Keyword(s):  
Iron Acquisition ◽  
Three Dimensional ◽  
Multidrug Resistant ◽  
New Drugs ◽  
Transport Systems ◽  
Resistant Bacteria ◽  
Gram Negative Bacteria ◽  
Structural Studies ◽  
Gram Negative ◽  
Novel Antibiotics

The emergence of multidrug-resistant Gram-negative bacteria such as P. aeruginosa has become a growing challenge for developing new drugs. One of the strategies recently used to improve the efficacy of the antibiotics is to increase their uptake by exploiting bacterial transport systems such as the siderophore-mediated iron acquisition system. BAL30072, a new monosulfactam conjugated to a siderophore moiety has been shown to have potent activity against many Gram-negative bacteria [1]. Several proteins affecting susceptibility to this antibiotic have been identified in Pseudomonas aeruginosa [2]. As part of the Translocation project (Innovative Medicines Initiative), we have undertaken structural studies of these proteins and have solved the three dimensional structures of two of these targets. The structure of PiuA, a TonB-dependent siderophore transporter involved in the uptake of siderophore antibiotics across the outer membrane [2,3] has been solved to a resolution of 1.9 Å. The structure of PiuC, an Fe(II)/α-ketoglutarate-dependent dioxygenase, has been solved to a resolution of 2.6 Å. The structural and biochemical studies of these proteins will help us to understand the mode of action of these novel antibiotics and subsequently help to design new drugs acting against multidrug-resistant bacteria.

scholarly journals Marine Actinobacteria: Screening for Predation Leads to the Discovery of Potential New Drugs against Multidrug-Resistant Bacteria

Antibiotics ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 91 ◽  
Author(s):  
Manar Ibrahimi ◽  
Wassila Korichi ◽  
Mohamed Hafidi ◽  
Laurent Lemee ◽  
Yedir Ouhdouch ◽  
...  
Keyword(s):  
Micrococcus Luteus ◽  
Multidrug Resistant ◽  
New Drugs ◽  
Resistant Bacteria ◽  
Clear Correlation ◽  
Growth Media ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Methicillin Resistant ◽  
Predatory Bacteria

Predatory bacteria constitute a heterogeneous group of prokaryotes able to lyse and feed on the cellular constituents of other bacteria in conditions of nutrient scarcity. In this study, we describe the isolation of Actinobacteria predator of other bacteria from the marine water of the Moroccan Atlantic coast. Only 4 Actinobacteria isolates showing strong predation capability against native or multidrug-resistant Gram-positive or Gram-negative bacteria were identified among 142 isolated potential predatory bacteria. These actinobacterial predators were shown to belong to the Streptomyces genus and to inhibit the growth of various native or multidrug-resistant micro-organisms, including Micrococcus luteus, Staphylococcus aureus (native and methicillin-resistant), and Escherichia coli (native and ampicillin-resistant). Even if no clear correlation could be established between the antibacterial activities of the selected predator Actinobacteria and their predatory activity, we cannot exclude that some specific bio-active secondary metabolites were produced in this context and contributed to the killing and lysis of the bacteria. Indeed, the co-cultivation of Actinobacteria with other bacteria is known to lead to the production of compounds that are not produced in monoculture. Furthermore, the production of specific antibiotics is linked to the composition of the growth media that, in our co-culture conditions, exclusively consisted of the components of the prey living cells. Interestingly, our strategy led to the isolation of bacteria with interesting inhibitory activity against methicillin-resistant S. aureus (MRSA) as well as against Gram-negative bacteria.

scholarly journals A novel chemical biology and computational approach to expedite the discovery of new-generation polymyxins against life-threatening Acinetobacter baumannii

2021 ◽  
Author(s):  
Xukai Jiang ◽  
Nitin A. Patil ◽  
Mohammad A. K. Azad ◽  
Hasini Wickremasinghe ◽  
Heidi Yu ◽  
...  
Keyword(s):  
Public Health ◽  
Chemical Biology ◽  
Multidrug Resistant ◽  
Computational Approach ◽  
Gram Negative Bacteria ◽  
Global Public Health ◽  
Gram Negative ◽  
Life Threatening ◽  
New Generation ◽  
Novel Antibiotics

Multidrug-resistant Gram-negative bacteria have been an urgent threat to global public health. Novel antibiotics are desperately needed to combat these 'superbugs'.

scholarly journals A small-molecule inhibitor of BamA impervious to efflux and the outer membrane permeability barrier

2019 ◽  
Vol 116 (43) ◽  
pp. 21748-21757 ◽  
Author(s):  
Elizabeth M. Hart ◽  
Angela M. Mitchell ◽  
Anna Konovalova ◽  
Marcin Grabowicz ◽  
Jessica Sheng ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Small Molecule ◽  
Cytoplasmic Membrane ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Permeability Barrier ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Bam Complex ◽  
Induced Aggregation

The development of new antimicrobial drugs is a priority to combat the increasing spread of multidrug-resistant bacteria. This development is especially problematic in gram-negative bacteria due to the outer membrane (OM) permeability barrier and multidrug efflux pumps. Therefore, we screened for compounds that target essential, nonredundant, surface-exposed processes in gram-negative bacteria. We identified a compound, MRL-494, that inhibits assembly of OM proteins (OMPs) by the β-barrel assembly machine (BAM complex). The BAM complex contains one essential surface-exposed protein, BamA. We constructed a bamA mutagenesis library, screened for resistance to MRL-494, and identified the mutation bamAE470K. BamAE470K restores OMP biogenesis in the presence of MRL-494. The mutant protein has both altered conformation and activity, suggesting it could either inhibit MRL-494 binding or allow BamA to function in the presence of MRL-494. By cellular thermal shift assay (CETSA), we determined that MRL-494 stabilizes BamA and BamAE470K from thermally induced aggregation, indicating direct or proximal binding to both BamA and BamAE470K. Thus, it is the altered activity of BamAE470K responsible for resistance to MRL-494. Strikingly, MRL-494 possesses a second mechanism of action that kills gram-positive organisms. In microbes lacking an OM, MRL-494 lethally disrupts the cytoplasmic membrane. We suggest that the compound cannot disrupt the cytoplasmic membrane of gram-negative bacteria because it cannot penetrate the OM. Instead, MRL-494 inhibits OMP biogenesis from outside the OM by targeting BamA. The identification of a small molecule that inhibits OMP biogenesis at the cell surface represents a distinct class of antibacterial agents.

scholarly journals Membrane Protein Complex ExbB4-ExbD1-TonB1from Escherichia coli Demonstrates Conformational Plasticity

2015 ◽  
Vol 197 (11) ◽  
pp. 1873-1885 ◽  
Author(s):  
Aleksandr Sverzhinsky ◽  
Jacqueline W. Chung ◽  
Justin C. Deme ◽  
Lucien Fabre ◽  
Kristian T. Levey ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Iron Acquisition ◽  
Proton Translocation ◽  
Three Dimensional ◽  
Structural Data ◽  
Structural Features ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Content Type

ABSTRACTIron acquisition at the outer membrane (OM) of Gram-negative bacteria is powered by the proton motive force (PMF) of the cytoplasmic membrane (CM), harnessed by the CM-embedded complex of ExbB, ExbD, and TonB. Its stoichiometry, ensemble structural features, and mechanism of action are unknown. By panning combinatorial phage libraries, periplasmic regions of dimerization between ExbD and TonB were predicted. Using overexpression of full-length His6-taggedexbB-exbDand S-taggedtonB, we purified detergent-solubilized complexes of ExbB-ExbD-TonB fromEscherichia coli. Protein-detergent complexes of ∼230 kDa with a hydrodynamic radius of ∼6.0 nm were similar to previously purified ExbB4-ExbD2complexes. Significantly, they differed in electronegativity by native agarose gel electrophoresis. The stoichiometry was determined to be ExbB4-ExbD1-TonB1. Single-particle electron microscopy agrees with this stoichiometry. Two-dimensional averaging supported the phage display predictions, showing two forms of ExbD-TonB periplasmic heterodimerization: extensive and distal. Three-dimensional (3D) particle classification showed three representative conformations of ExbB4-ExbD1-TonB1. Based on our structural data, we propose a model in which ExbD shuttles a proton across the CM via an ExbB interprotein rearrangement. Proton translocation would be coupled to ExbD-mediated collapse of extended TonB in complex with ligand-loaded receptors in the OM, followed by repositioning of TonB through extensive dimerization with ExbD. Here we present the first report for purification of the ExbB-ExbD-TonB complex, molar ratios within the complex (4:1:1), and structural biology that provides insights into 3D organization.IMPORTANCEReceptors in the OM of Gram-negative bacteria allow entry of iron-bound siderophores that are necessary for pathogenicity. Numerous iron-acquisition strategies rely upon a ubiquitous and unique protein for energization: TonB. Complexed with ExbB and ExbD, the Ton system links the PMF to OM transport. Blocking iron uptake by targeting a vital nanomachine holds promise in therapeutics. Despite much research, the stoichiometry, structural arrangement, and molecular mechanism of the CM-embedded ExbB-ExbD-TonB complex remain unreported. Here we demonstratein vitroevidence of ExbB4-ExbD1-TonB1complexes. Using 3D EM, we reconstructed the complex in three conformational states that show variable ExbD-TonB heterodimerization. Our structural observations form the basis of a model for TonB-mediated iron acquisition.

scholarly journals Species Diversity of Environmental GIM-1-Producing Bacteria Collected during a Long-Term Outbreak

2016 ◽  
Vol 82 (12) ◽  
pp. 3605-3610 ◽  
Author(s):  
Andreas F. Wendel ◽  
Sofija Ressina ◽  
Susanne Kolbe-Busch ◽  
Klaus Pfeffer ◽  
Colin R. MacKenzie
Keyword(s):  
Resistance Genes ◽  
Multidrug Resistant ◽  
Hospital Environment ◽  
Resistant Bacteria ◽  
Environmental Sampling ◽  
Gram Negative Bacteria ◽  
Multidrug Resistant Bacteria ◽  
Gram Negative ◽  
Content Type

ABSTRACTReports of outbreaks concerning carbapenemase-producing Gram-negative bacteria in which the main source of transmission is the hospital environment are increasing. This study describes the results of environmental sampling in a protracted polyspecies metallo-beta-lactamase GIM-1 outbreak driven by plasmids and bacterial clones ofEnterobacter cloacaeandPseudomonas aeruginosain a tertiary care center. Environmental sampling targeting wet locations (especially sinks) was carried out on a surgical intensive care unit and on a medical ward on several occasions in 2012 and 2013. We were able to demonstrate 43blaGIM-1-carrying bacteria (mainly nonfermenters but alsoEnterobacteriaceae) that were either related or unrelated to clinical strains in 30 sinks and one hair washbasin. GIM-1 was found in 12 different species, some of which are described here as carriers of GIM-1. Forty out of 43 bacteria displayed resistance to carbapenems and, in addition, to various non-beta-lactam antibiotics. Colistin resistance was observed in twoE. cloacaeisolates with MICs above 256 mg/liter. TheblaGIM-1gene was harbored in 12 different class 1 integrons, some without the typical 3′ end. TheblaGIM-1gene was localized on plasmids in five isolates.In vitroplasmid transfer by conjugation was successful in one isolate. The environment, with putatively multispecies biofilms, seems to be an important biological niche for multidrug-resistant bacteria and resistance genes. Biofilms may serve as a “melting pot” for horizontal gene transfer, for dissemination into new species, and as a reservoir to propagate future hospital outbreaks.IMPORTANCEIn Gram-negative bacteria, resistance to the clinically relevant broad-spectrum carbapenem antibiotics is a major public health concern. Major reservoirs for these resistant organisms are not only the gastrointestinal tracts of animals and humans but also the (hospital) environment. Due to the difficulty in eradicating biofilm formation in the latter, a sustained dissemination of multidrug-resistant bacteria from the environment can occur. In addition, horizontal transfer of resistance genes on mobile genetic elements within biofilms adds to the total “resistance gene pool” in the environment. To gain insight into the transmission pathways of a rare and locally restricted carbapenemases resistance gene (blaGIM-1), we analyzed the genetic background of theblaGIM-1gene in environmental bacteria during a long-term polyspecies outbreak in a German hospital.

scholarly journals Antibiotic Resistant Bacterial Isolates from Captive Green Turtles andIn VitroSensitivity to Bacteriophages

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Alessandro Delli Paoli Carini ◽  
Ellen Ariel ◽  
Jacqueline Picard ◽  
Lisa Elliott
Keyword(s):  
Rapid Evolution ◽  
Chelonia Mydas ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Green Turtles ◽  
Resistant Genes ◽  
Resistance Rates

This study aimed to test multidrug resistant isolates from hospitalised green turtles(Chelonia mydas)and their environment in North Queensland, Australia, forin vitrosusceptibility to bacteriophages. Seventy-one Gram-negative bacteria were isolated from green turtle eye swabs and water samples. Broth microdilution tests were used to determine antibiotic susceptibility. All isolates were resistant to at least two antibiotics, with 24% being resistant to seven of the eight antibiotics. Highest resistance rates were detected to enrofloxacin (77%) and ampicillin (69.2%). More than 50% resistance was also found to amoxicillin/clavulanic acid (62.5%), ceftiofur (53.8%), and erythromycin (53.3%). All the enriched phage filtrate mixtures resulted in the lysis of one or more of the multidrug resistant bacteria, includingVibrio harveyiandV. parahaemolyticus. These results indicate that antibiotic resistance is common in Gram-negative bacteria isolated from hospitalised sea turtles and their marine environment in North Queensland, supporting global concern over the rapid evolution of multidrug resistant genes in the environment. Using virulent bacteriophages as antibiotic alternatives would not only be beneficial to turtle health but also prevent further addition of multidrug resistant genes to coastal waters.

scholarly journals Trends of Bloodstream Infections in a University Greek Hospital during a Three-Year Period: Incidence of Multidrug-Resistant Bacteria and Seasonality in Gram-negative Predominance

2017 ◽  
Vol 66 (2) ◽  
pp. 171-180 ◽  
Author(s):  
Fevronia Kolonitsiou ◽  
Matthaios Papadimitriou-Olivgeris ◽  
Anastasia Spiliopoulou ◽  
Vasiliki Stamouli ◽  
Vasileios Papakostas ◽  
...  
Keyword(s):  
Bloodstream Infections ◽  
Multidrug Resistant ◽  
Blood Cultures ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Gram Negative Bacteria ◽  
Multidrug Resistant Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
Carbapenem Resistant

The aim of the study was to assess the epidemiology, the incidence of multidrug-resistant bacteria and bloodstream infections’ (BSIs) seasonality in a university hospital. This retrospective study was carried out in the University General Hospital of Patras, Greece, during 2011–13 y. Blood cultures from patients with clinical presentation suggestive of bloodstream infection were performed by the BacT/ALERT System. Isolates were identified by Vitek 2 Advanced Expert System. Antibiotic susceptibility testing was performed by the disk diffusion method and E-test. Resistance genes (mecA in staphylococci; vanA/vanB/vanC in enterococci; blaKPC/blaVIM/blaNDM in Klebsiella spp.) were detected by PCR. In total, 4607 (9.7%) blood cultures were positive from 47451 sets sent to Department of Microbiology, representing 1732 BSIs. Gram-negative bacteria (52.3%) were the most commonly isolated, followed by Gram-positive (39.5%), fungi (6.6%) and anaerobes bacteria (1.8%). The highest contamination rate was observed among Gram-positive bacteria (42.3%). Among 330 CNS and 150 Staphylococcus aureus, 281 (85.2%) and 60 (40.0%) were mecA-positive, respectively. From 113 enterococci, eight were vanA, two vanB and two vanC-positives. Of the total 207 carbapenem-resistant Klebsiella pneumoniae (73.4%), 202 carried blaKPC, four blaKPC and blaVIM and one blaVIM. A significant increase in monthly BSIs’ incidence was shown (R2: 0.449), which may be attributed to a rise of Gram-positive BSIs (R2: 0.337). Gram-positive BSIs were less frequent in spring (P < 0.001), summer (P < 0.001), and autumn (P < 0.001), as compared to winter months, while Gram-negative bacteria (P < 0.001) and fungi (P < 0.001) were more frequent in summer months. BSIs due to methicillin resistant S. aureus and carbapenem-resistant Gram-negative bacteria increased during the study period. The increasing incidence of BSIs can be attributed to an increase of Gram-positive BSI incidence, even though Gram-negative bacteria remained the predominant ones. Seasonality may play a role in the predominance of Gram-negative’s BSI.

scholarly journals Antimicrobial Resistance in Hospitalized Surgical Patients: a Silently Emerging Public Health Concern in Benin.

2020 ◽  
Author(s):  
Carine Laurence YEHOUENOU ◽  
Arsène A. KPANGON ◽  
Dissou AFFOLABI ◽  
Hector RODRIGUEZ-VILLALOBOS ◽  
Françoise Van Bambeke ◽  
...  
Keyword(s):  
Gastrointestinal Surgery ◽  
Surgical Site Infections ◽  
Multidrug Resistant ◽  
Public Hospitals ◽  
Health Concern ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Gram Negative Bacteria ◽  
Strict Adherence ◽  
Gram Negative

Abstract Background: Surgical site infections are related to high morbidity, mortality and healthcare costs. As the emergence of multidrug-resistant bacterial pathogens in hospitals is becoming a worldwide challenge for surgeons who treat healthcare-associated infections, we wished to identify the causative agents involved in surgical site infections and their susceptibility pattern in six public hospitals in Benin. Methods: Using standard microbiological procedures, we processed pus specimens collected from obstetrics and gastrointestinal surgery wards. Mass spectrometry (MALDI-TOF) was used for confirmation. The antibiotic susceptibility test firstly used the Kirby-Bauer disc diffusion method. The secondary test by microdilution used the Beckton Dickinson Phoenix automated system (Becton Dickinson Diagnostic, USA). Results: We included 304 patients (mean age 32 ± 11 years), whose median length of stay was 9 days. A total of 259 wound swabs (85.2%) had positive aerobic bacterial growth. In obstetrics S. aureus (28.5%, n=42) was the most common isolate. In contrast, Gram-negative bacteria (GNB) were predominant in gastrointestinal surgery. The most dominant being E.coli (38.4%, n=31). Overall, 90.8% (n=208) of aerobic bacteria were multidrug resistant. Two-third of S. aureus (65.3%, n= 32) were methicillin-resistant Staphylococcus aureus (MRSA), three of which carried both MRSA and induced clindamycin resistance (ICR). GNB showed high resistance to ceftazidime, ceftriaxone and cefepime. Extended-spectrum beta-lactamases were presented by 69.4% of E.coli (n=43/62) and 83.3% of K. pneumoniae (n=25/30). Overall, twelve Gram negative bacteria (5.24%) isolates showed resistance to at least one carbapenem. No isolates showed a wild-type susceptible phenotype.Conclusion: This study shows the alarming prevalence of multidrug resistant organisms from surgical site infections in Benin hospitals. To reduce the spread of these multidrug-resistant bacteria, periodic surveillance of surgical site infections and strict adherence to good hand-hygiene practice are essential.

scholarly journals Detection of Multidrug Resistant Gram Negative Bacteria in Healthy Cattle from Maiduguri Metropolitan, Nigeria

2020 ◽  
pp. 68-76
Author(s):  
Adam Mustapha ◽  
Mustafa Alhaji Isa ◽  
Ibrahim Yusuf Ngoshe ◽  
Hashidu Bala
Keyword(s):  
Antibiotic Resistance ◽  
Clinical Laboratory ◽  
Resistance Index ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Gram Negative Bacteria ◽  
Salmonella Spp ◽  
Gram Negative ◽  
Healthy Cattle

Aim: Prevalence of multidrug resistant bacteria on apparently health animals has turned antibiotic resistance to multifaceted process and threatens global food security and public health. The aim of the present study was to investigate the resistance profile of isolates from apparently healthy cattle in Maiduguri, Nigeria. Methodology: A total of 120 nasal swab samples were collected from cattle. Colony identification was according to the guidelines of Bergey’s Manual of Determinative Bacteriology. The susceptibility pattern of the isolates was conducted on the identified isolates according to the Modified Kirby-Baur disc diffusion method on Muller-Hilton agar and interpreted according to the procedures of Clinical Laboratory Standards Institute (CLSI, 2018) guidelines. Multiple Antibiotic Resistance Index (MARI) was calculated using the formula, MARI=a/b where “a” is the number of antibiotic resisted and “b” is the total number of antibiotic used in the study. Results: Of the total samples (120) from cattle 96 (80%) detected the following isolates; E. coli was the most commonly recovered isolates (33, 34.4%), followed by Klebsiella spp (28, 29.2%), Salmonella spp (21, 21.9%) and Pseudomonas aeruginosa (14, 14.5%). In this study, all the recovered isolates were found to be multidrug resistant gram negative bacteria, with highest resistance was shown by Salmonella spp. The high MARI observed in all the isolates in this study ranging from 0.7 to 0.9. MARI value of 0.2 > is suggests multiple antibiotic resistant bacteria and indicate presence of highly resistant bacteria. Conclusion: The study indicates highly resistant bacteria are carried by healthy food animals. Thus, there is need for continued monitoring of antibiotics use in animal husbandry to prevent further spread of resistance in Maiduguri, Nigeria.

Oral, nasopharyngeal, and gut decontamination in the ICU

2016 ◽  
Author(s):  
Evelien Oostdijk ◽  
Marc Bonten
Keyword(s):  
Digestive Tract ◽  
Antibiotic Use ◽  
Multidrug Resistant ◽  
Selective Decontamination ◽  
Resistant Bacteria ◽  
Gram Negative Bacteria ◽  
Detailed Data ◽  
Multidrug Resistant Bacteria ◽  
Gram Negative ◽  
Selective Oropharyngeal Decontamination

Many infections are caused by enteric bacilli, presumably from endogenous origin. Selective decontamination of the digestive tract (SDD) was developed to selectively eliminate the aerobic Gram-negative bacilli from the digestive tract, leaving the anaerobic flora unaffected. As an alternative to SDD, investigators have evaluated the effects of selective oropharyngeal decontamination (SOpD) alone. Most detailed data on the effects of SDD and SOpD in ICU-patients come from two studies performed in Dutch ICUs. The Dutch studies provide strong evidence that SDD and SOpD reduce ICUmortality, ICU-acquired bacteraemia with Gram-negative bacteria, and systemic antibiotic use. Although successful application has been reported from several solitary ICUs across Europe, it is currently unknown to what extent these effects can be achieved in settings with different bacterial ecology. More studies are needed on the use of SDD or SOpD as a measure to control outbreaks with multidrug resistant bacteria.

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