scholarly journals Bioprospecting of Native Efflux Pumps To Enhance Furfural Tolerance in EthanologenicEscherichia coli

2019 ◽  
Vol 85 (6) ◽  
Author(s):  
Gavin Kurgan ◽  
Larry A. Panyon ◽  
Yesenia Rodriguez-Sanchez ◽  
Eric Pacheco ◽  
Lizbeth M. Nieves ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Cost Effective ◽  
Efflux Pumps ◽  
Toxic Chemicals ◽  
Screening Methods ◽  
Microbial Conversion ◽  
Content Type ◽  
Furfural Tolerance ◽  
Plasmid Library ◽  
Lignocellulose Conversion

ABSTRACTEfficient microbial conversion of lignocellulose into valuable products is often hindered by the presence of furfural, a dehydration product of pentoses in hemicellulose sugar syrups derived from woody biomass. For a cost-effective lignocellulose microbial conversion, robust biocatalysts are needed that can tolerate toxic inhibitors while maintaining optimal metabolic activities. A comprehensive plasmid-based library encoding native multidrug resistance (MDR) efflux pumps, porins, and select exporters fromEscherichia coliwas screened for furfural tolerance in an ethanologenicE. colistrain. Small multidrug resistance (SMR) pumps, such as SugE and MdtJI, as well as a lactate/glycolate:H+symporter, LldP, conferred furfural tolerance in liquid culture tests. Expression of the SMR pump potentially increased furfural efflux and cellular viability upon furfural assault, suggesting novel activities for SMR pumps as furfural efflux proteins. Furthermore, induced expression ofmdtJIenhanced ethanol fermentative production of LY180 in the presence of furfural or 5-hydroxymethylfurfural, further demonstrating the applications of SMR pumps. This work describes an effective approach to identify useful efflux systems with desired activities for nonnative toxic chemicals and provides a platform to further enhance furfural efflux by protein engineering and mutagenesis.IMPORTANCELignocellulosic biomass, especially agricultural residues, represents an important potential feedstock for microbial production of renewable fuels and chemicals. During the deconstruction of hemicellulose by thermochemical processes, side products that inhibit cell growth and production, such as furan aldehydes, are generated, limiting cost-effective lignocellulose conversion. Here, we developed a new approach to increase cellular tolerance by expressing multidrug resistance (MDR) pumps with putative efflux activities for furan aldehydes. The developed plasmid library and screening methods may facilitate new discoveries of MDR pumps for diverse toxic chemicals important for microbial conversion.

scholarly journals Bifunctional Enzyme SpoT Is Involved in Biofilm Formation ofHelicobacter pyloriwith Multidrug Resistance by Upregulating Efflux Pump Hp1174 (gluP)

2018 ◽  
Vol 62 (11) ◽  
Author(s):  
Xiaoran Ge ◽  
Yuying Cai ◽  
Zhenghong Chen ◽  
Sizhe Gao ◽  
Xiwen Geng ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Multidrug Resistant ◽  
Major Facilitator Superfamily ◽  
Bifunctional Enzyme ◽  
Content Type ◽  
H Pylori ◽  
Major Facilitator

ABSTRACTThe drug resistance ofHelicobacter pyloriis gradually becoming a serious problem. Biofilm formation is an important factor that leads to multidrug resistance (MDR) in bacteria. The ability ofH. pylorito form biofilms on the gastric mucosa is known. However, there are few studies on the regulatory mechanisms ofH. pyloribiofilm formation and multidrug resistance. Guanosine 3′-diphosphate 5′-triphosphate and guanosine 3′,5′-bispyrophosphate [(p)ppGpp] are global regulatory factors and are synthesized inH. pyloriby the bifunctional enzyme SpoT. It has been reported that (p)ppGpp is involved in the biofilm formation and multidrug resistance of various bacteria. In this study, we found that SpoT also plays an important role inH. pyloribiofilm formation and multidrug resistance. Therefore, it was necessary to carry out some further studies regarding its regulatory mechanism. Considering that efflux pumps are of great importance in the biofilm formation and multidrug resistance of bacteria, we tried to determine whether efflux pumps controlled by SpoT participate in these activities. We found that Hp1174 (glucose/galactose transporter [gluP]), an efflux pump of the major facilitator superfamily (MFS), is highly expressed in biofilm-forming and multidrug-resistant (MDR)H. pyloristrains and is upregulated by SpoT. Through further research, we determined thatgluPis involved inH. pyloribiofilm formation and multidrug resistance. Furthermore, the average expression level ofgluPin the clinical MDR strains (C-MDR) was considerably higher than that in the clinical drug-sensitive strains (C-DSS). Taken together, our results revealed a novel molecular mechanism ofH. pyloriresistance to multidrug exposure.

scholarly journals Biolog Phenotype Microarray Is a Tool for the Identification of Multidrug Resistance Efflux Pump Inducers

2018 ◽  
Vol 62 (11) ◽  
Author(s):  
P. Blanco ◽  
F. Corona ◽  
J. L. Martínez
Keyword(s):  
Multidrug Resistance ◽  
Fluorescent Protein ◽  
Efflux Pump ◽  
Yellow Fluorescent Protein ◽  
Efflux Pumps ◽  
Phenotype Microarray ◽  
Content Type ◽  
Environmental Signals ◽  
Reverse Transcription Pcr ◽  
Biolog Phenotype Microarray

ABSTRACTMultidrug resistance efflux pumps frequently present low levels of basal expression. However, antibiotic-resistant mutants that overexpress these resistance determinants are selected during infection. In addition, increased expression of efflux pumps can be induced by environmental signals/cues, which can lead to situations of transient antibiotic resistance. In this study, we have applied a novel high-throughput methodology in order to identify inducers able to trigger the expression of theStenotrophomonas maltophiliaSmeVWX and SmeYZ efflux pumps. To that end, bioreporters in which the expression of the yellow fluorescent protein (YFP) is linked to the activity of eithersmeVWXorsmeYZpromoters were developed and used for the screening of potential inducers of the expression of these efflux pumps using Biolog phenotype microarrays. YFP production was also measured by flow cytometry, and the levels of expression ofsmeVandsmeYin the presence of a set of selected compounds were also determined by real-time reverse transcription-PCR (RT-PCR). The expression ofsmeVWXwas induced by iodoacetate, clioquinol, and selenite, while boric acid, erythromycin, chloramphenicol, and lincomycin triggeredsmeYZexpression. The susceptibility to antibiotics that are known substrates of the efflux pumps decreased in the presence of the inducers. However, the analyzed multidrug efflux systems did not contribute toS. maltophiliaresistance to the studied inducers. To sum up, the use of fluorescent bioreporters in combination with Biolog plates is a valuable tool for identifying inducers of the expression of bacterial multidrug resistance efflux pumps, and likely of other bacterial systems whose expression is regulated in response to signals/cues.

scholarly journals Detection of Carbapenemase Production in a Collection of Enterobacteriaceae with Characterized Resistance Mechanisms from Clinical and Environmental Origins by Use of Both Carba NP and Blue-Carba Tests: TABLE 1

2015 ◽  
Vol 54 (2) ◽  
pp. 464-466 ◽  
Author(s):  
Sergio García-Fernández ◽  
María-Isabel Morosini ◽  
Desirèe Gijón ◽  
Lorena Beatobe ◽  
Patricia Ruiz-Garbajosa ◽  
...  
Keyword(s):  
High Sensitivity ◽  
Cost Effective ◽  
Multidrug Resistant ◽  
Resistance Mechanisms ◽  
Rapid Screening ◽  
Resistant Bacteria ◽  
Screening Methods ◽  
Routine Laboratory ◽  
Multidrug Resistant Bacteria ◽  
Content Type

Rapid-screening methods to confirm the presence of resistance mechanisms in multidrug-resistant bacteria are currently recommended. Carba NP and Blue-Carba tests were evaluated in carbapenemase-producingEnterobacteriaceaefrom hospital (n= 102) and environmental (n= 57) origins for detecting the different molecular classes among them. Both methods showed to be fast and cost-effective, with high sensitivity (98% to 100%) and specificity (100%), and may be easily introduced in the routine laboratory.

scholarly journals Riboswitch-Associated Guanidinium-Selective Efflux Pumps Frequently Transmitted on Proteobacterial Plasmids Increase Escherichia coli Biofilm Tolerance to Disinfectants

2020 ◽  
Vol 202 (23) ◽  
Author(s):  
Carmine J. Slipski ◽  
Taylor R. Jamieson ◽  
George G. Zhanel ◽  
Denice C. Bay
Keyword(s):  
Escherichia Coli ◽  
Multidrug Resistance ◽  
Quaternary Ammonium ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Cloning And Expression ◽  
Upstream Region ◽  
Ammonium Compound ◽  
Content Type ◽  
Small Multidrug Resistance

ABSTRACT Members of the small multidrug resistance (SMR) efflux pump family known as SugE (recently renamed Gdx) are known for their narrow substrate selectivity to small guanidinium (Gdm+) compounds and disinfectant quaternary ammonium compounds (QACs). Gdx members have been identified on multidrug resistance plasmids in Gram-negative bacilli, but their functional role remains unclear, as few have been characterized. Here, we conducted a survey of sequenced proteobacterial plasmids that encoded one or more SugE/Gdx sequences in an effort to (i) identify the most frequently represented Gdx member(s) on these plasmids and their sequence diversity, (ii) verify if Gdx sequences possess a Gdm+ riboswitch that regulates their translation similarly to chromosomally encoded Gdx members, and (iii) determine the antimicrobial susceptibility profile of the most predominate Gdx member to various QACs and antibiotics in Escherichia coli strains BW25113 and KAM32. The results of this study determined 14 unique SugE sequences, but only one Gdx sequence, annotated as “SugE(p),” predominated among the >140 plasmids we surveyed. Enterobacterales plasmids carrying sugE(p) possessed a guanidine II riboswitch similar to the upstream region of E. coli gdx. Cloning and expression of sugE(p), gdx, and emrE sequences into a low-copy-number expression vector (pMS119EH) revealed significant increases in QAC resistance to a limited range of detergent-like QACs only when gdx and sugE(p) transformants were grown as biofilms. These findings suggest that sugE(p) presence on proteobacterial plasmids may be driven by species that frequently encounter Gdm+ and QAC exposure. IMPORTANCE This study characterized the function of antimicrobial-resistant phenotypes attributed to plasmid-encoded guanidinium-selective small multidrug resistance (Gdm/SugE) efflux pumps. These sequences are frequently monitored as biocide resistance markers in antimicrobial resistance surveillance studies. Our findings reveal that enterobacterial gdm sequences transmitted on plasmids possess a guanidine II riboswitch, which restricts transcript translation in the presence of guanidinium. Cloning and overexpression of this gdm sequence revealed that it confers higher resistance to quaternary ammonium compound (QAC) disinfectants (which possess guanidium moieties) when grown as biofilms. Since biofilms are commonly eradicated with QAC-containing compounds, the presence of this gene on plasmids and its biofilm-specific resistance are a growing concern for clinical and food safety prevention measures.

The cost-effectiveness of insulin analogs and regular insulin for diabetes control: a case study in Iran

2020 ◽  
Vol 33 (4/5) ◽  
pp. 323-331
Author(s):  
Mohsen pakdaman ◽  
Raheleh akbari ◽  
Hamid reza Dehghan ◽  
Asra Asgharzadeh ◽  
Mahdieh Namayandeh
Keyword(s):  
Cost Effectiveness ◽  
Cost Effective ◽  
Regular Insulin ◽  
Diabetes Control ◽  
Diabetic Patients ◽  
Insulin Analogs ◽  
Content Type ◽  
Patients With Diabetes ◽  
The Government ◽  
The Cost

PurposeFor years, traditional techniques have been used for diabetes treatment. There are two major types of insulin: insulin analogs and regular insulin. Insulin analogs are similar to regular insulin and lead to changes in pharmacokinetic and pharmacodynamic properties. The purpose of the present research was to determine the cost-effectiveness of insulin analogs versus regular insulin for diabetes control in Yazd Diabetes Center in 2017.Design/methodology/approachIn this descriptive–analytical research, the cost-effectiveness index was used to compare insulin analogs and regular insulin (pen/vial) for treatment of diabetes. Data were analyzed in the TreeAge Software and a decision tree was constructed. A 10% discount rate was used for ICER sensitivity analysis. Cost-effectiveness was examined from a provider's perspective.FindingsQALY was calculated to be 0.2 for diabetic patients using insulin analogs and 0.05 for those using regular insulin. The average cost was $3.228 for analog users and $1.826 for regular insulin users. An ICER of $0.093506/QALY was obtained. The present findings suggest that insulin analogs are more cost-effective than regular insulin.Originality/valueThis study was conducted using a cost-effectiveness analysis to evaluate insulin analogs versus regular insulin in controlling diabetes. The results of study are helpful to the government to allocate more resources to apply the cost-effective method of the treatment and to protect patients with diabetes from the high cost of treatment.

scholarly journals Characterization of RarA, a Novel AraC Family Multidrug Resistance Regulator in Klebsiella pneumoniae

2012 ◽  
Vol 56 (8) ◽  
pp. 4450-4458 ◽  
Author(s):  
Mark Veleba ◽  
Paul G. Higgins ◽  
Gerardo Gonzalez ◽  
Harald Seifert ◽  
Thamarai Schneiders
Keyword(s):  
Multidrug Resistance ◽  
Klebsiella Pneumoniae ◽  
Efflux Pump ◽  
Content Type ◽  
Resistance Phenotype ◽  
Serratia Proteamaculans ◽  
Virulence Potential ◽  
Article Type ◽  
Acrab Efflux Pump

ABSTRACTTranscriptional regulators, such as SoxS, RamA, MarA, and Rob, which upregulate the AcrAB efflux pump, have been shown to be associated with multidrug resistance in clinically relevant Gram-negative bacteria. In addition to the multidrug resistance phenotype, these regulators have also been shown to play a role in the cellular metabolism and possibly the virulence potential of microbial cells. As such, the increased expression of these proteins is likely to cause pleiotropic phenotypes.Klebsiella pneumoniaeis a major nosocomial pathogen which can express the SoxS, MarA, Rob, and RamA proteins, and the accompanying paper shows that the increased transcription oframAis associated with tigecycline resistance (M. Veleba and T. Schneiders, Antimicrob. Agents Chemother. 56:4466–4467, 2012). Bioinformatic analyses of the availableKlebsiellagenome sequences show that an additional AraC-type regulator is encoded chromosomally. In this work, we characterize this novel AraC-type regulator, hereby called RarA (Regulator of antibiotic resistance A), which is encoded inK. pneumoniae,Enterobactersp. 638,Serratia proteamaculans568, andEnterobacter cloacae. We show that the overexpression ofrarAresults in a multidrug resistance phenotype which requires a functional AcrAB efflux pump but is independent of the other AraC regulators. Quantitative real-time PCR experiments show thatrarA(MGH 78578 KPN_02968) and its neighboring efflux pump operonoqxAB(KPN_02969_02970) are consistently upregulated in clinical isolates collected from various geographical locations (Chile, Turkey, and Germany). Our results suggest thatrarAoverexpression upregulates theoqxABefflux pump. Additionally, it appears thatoqxR, encoding a GntR-type regulator adjacent to theoqxABoperon, is able to downregulate the expression of theoqxABefflux pump, where OqxR complementation resulted in reductions to olaquindox MICs.

Job crafting, workplace civility and work outcomes: the mediating role of work engagement

2020 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Anuj Gupta ◽  
Pankaj Singh
Keyword(s):  
Life Satisfaction ◽  
Work Engagement ◽  
Cost Effective ◽  
Job Crafting ◽  
Intention To Quit ◽  
Work Outcomes ◽  
Equation Modeling ◽  
Content Type ◽  
General Life Satisfaction ◽  
Workplace Civility

Purpose The antecedents and consequences of work engagement have been extensively discussed and analyzed in the previous literature; however, identifying cost-effective measures that can sustain work engagement to boost work outcomes has received sparse attention in the Indian information technology (IT) context. This study aims to provide new insights concerning the associations of job crafting and workplace civility with work engagement and its corresponding outcomes, such as change perception, general life satisfaction and intention to quit. Design/methodology/approach Hypotheses were tested using structural equation modeling on data obtained from 369 software developers in India using questionnaire surveys. Findings Results confirmed that work engagement partially mediated the association of job crafting and workplace civility with the perception of change and general life satisfaction. The negative associations of job crafting and workplace civility with the intention to quit were also partially mediated by work engagement. The findings can be used to inform human resources strategies to boost work engagement and subsequent work outcomes. Research limitations/implications The results of this empirical work will offer insights to managers who are looking for cost-effective interventions and behaviors aimed at increasing work engagement and, consequently, achieving effective work outcomes. Originality/value This study contributes by empirically testing the application of novel employee-driven practices in improving work engagement and work outcomes, particularly in the context of IT companies in India.

scholarly journals Phosphorylation of MgrA and Its Effect on Expression of the NorA and NorB Efflux Pumps of Staphylococcus aureus

2010 ◽  
Vol 192 (10) ◽  
pp. 2525-2534 ◽  
Author(s):  
Que Chi Truong-Bolduc ◽  
David C. Hooper
Keyword(s):  
Staphylococcus Aureus ◽  
Multidrug Resistance ◽  
Virulence Factors ◽  
Double Mutant ◽  
Efflux Pumps ◽  
Efflux Transporters ◽  
Global Regulator ◽  
Binding Assays ◽  
Genes Encoding ◽  
Gel Shift

ABSTRACT MgrA is a global regulator in Staphylococcus aureus that controls the expression of diverse genes encoding virulence factors and multidrug resistance (MDR) efflux transporters. We identified pknB, which encodes the (Ser/Thr) kinase PknB, in the S. aureus genome. PknB was able to autophosphorylate as well as phosphorylate purified MgrA. We demonstrated that rsbU, which encodes a Ser/Thr phosphatase and is involved in the activation of the SigB regulon, was able to dephosphorylate MgrA-P but not PknB-P. Serines 110 and 113 of MgrA were found to be phosphorylated, and Ala substitutions at these positions resulted in reductions in the level of phosphorylation of MgrA. DNA gel shift binding assays using norA and norB promoters showed that MgrA-P was able to bind the norB promoter but not the norA promoter, a pattern which was the reverse of that for unphosphorylated MgrA. The double mutant MgrAS110A-S113A bound to the norA promoter but not the norB promoter. The double mutant led to a 2-fold decrease in norA transcripts and a 2-fold decrease in the MICs of norfloxacin and ciprofloxacin in strain RN6390. Thus, phosphorylation of MgrA results in loss of binding to the norA promoter, but with a gain of the ability to bind the norB promoter. Loss of the ability to phosphorylate MgrA by Ala substitution resulted in increased repression of norA expression and in reductions in susceptibilities to NorA substrates.

A revised FMEA application to the quality control management

2014 ◽  
Vol 31 (7) ◽  
pp. 788-810 ◽  
Author(s):  
Claudia Paciarotti ◽  
Giovanni Mazzuto ◽  
Davide D’Ettorre
Keyword(s):  
Quality Control ◽  
Failure Modes ◽  
Cost Effective ◽  
Weight Functions ◽  
Time Saving ◽  
Content Type ◽  
Complete Procedure ◽  
Medium Enterprise ◽  
Control Management ◽  
Critical Situations

Purpose – The purpose of this paper is to propose a cost-effective, time-saving and easy-to-use failure modes and effects analysis (FMEA) system applied on the quality control of supplied products. The traditional FMEA has been modified and adapted to fit the quality control features and requirements. The paper introduces a new and revised FMEA approach, where the “failure concept” has been modified with “defect concept.” Design/methodology/approach – The typical FMEA parameters have been modified, and a non-linear scale has been introduced to better evaluate the FMEA parameters. In addition, two weight functions have been introduced in the risk priority number (RPN) calculus in order to consider different critical situations previously ignored and the RPN is assigned to several similar products in order to reduce the problem of complexity. Findings – A complete procedure is provided in order to assist managers in deciding on the critical suppliers, the creation of homogeneous families overcome the complexity of single product code approach, in RPN definition the relative importance of factors is evaluated. Originality/value – This different approach facilitates the quality control managers acting as a structured and “friendly” decision support system: the quality control manager can easily evaluate the critical situations and simulate different scenarios of corrective actions in order to choose the best one. This FMEA technique is a dynamic tool and the performed process is an iterative one. The method has been applied in a small medium enterprise producing hydro massage bathtub, shower, spas and that commercializes bathroom furniture. The firm application has been carried out involving a cross-functional and multidisciplinary team.

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