Strategies for Rapid Identification of Acinetobacter baumannii Membrane Proteins and Polymyxin B’s Effects

Acinetobacter baumannii, especially multidrug resistant Acinetobacter baumannii, is a notable source of pressure in the areas of public health and antibiotic development. To overcome this problem, attention has been focused on membrane proteins. Different digestion methods and extraction detergents were examined for membrane proteome sample preparation, and label-free quantitative and targeted proteome analyses of the polymyxin B-induced Acinetobacter baumannii ATCC 19606 membrane proteome were performed based on nano LC-MS/MS. Ultracentrifugation of proteins at a speed of 150,000×g, digestion by trypsin, filter-aided sample preparation, and detergents such as lauryldimethylamine-N-oxide were proved as a fast and effective way for identification of membrane proteome by nano LC-MS/MS. Upon treatment with polymyxin B, expression levels of 15 proteins related to membrane structure, transporters, cell surface, and periplasmic space were found to be significantly changed. Furthermore, targeted proteome was also used to confirm these changes. A relatively rapid membrane proteome preparation method was developed, and a more comprehensive view of changes in the Acinetobacter baumannii membrane proteome under polymyxin B pressure was obtained.

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Comparative metabolomics reveals key pathways associated with the synergistic activity of polymyxin B and rifampicin combination against multidrug-resistant Acinetobacter baumannii

Biochemical Pharmacology ◽

10.1016/j.bcp.2020.114400 ◽

2021 ◽

Vol 184 ◽

pp. 114400

Author(s):

Jinxin Zhao ◽

Mei-Ling Han ◽

Yan Zhu ◽

Yu-Wei Lin ◽

Yi-Wen Wang ◽

...

Keyword(s):

Acinetobacter Baumannii ◽

Polymyxin B ◽

Multidrug Resistant ◽

Synergistic Activity ◽

Comparative Metabolomics ◽

Key Pathways

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Expression, purification, crystallization and preliminary crystallographic analysis of the phosphoglycerate kinase fromAcinetobacter baumannii

Acta Crystallographica Section F Structural Biology and Crystallization Communications ◽

10.1107/s1744309112020222 ◽

2012 ◽

Vol 68 (7) ◽

pp. 790-792

Author(s):

Kayla Baretta ◽

Craig Garen ◽

Jiang Yin ◽

Michael N. G. James

Keyword(s):

Synchrotron Radiation ◽

Acinetobacter Baumannii ◽

Light Source ◽

Phosphoglycerate Kinase ◽

Multidrug Resistant ◽

Crystallographic Analysis ◽

Lithium Sulfate ◽

Space Group ◽

Antibiotic Development ◽

Potential Target

Acinetobacter baumanniiis a common multidrug-resistant clinical pathogen that is often found in hospitals. TheA. baumanniiphosphoglycerate kinase (AbPGK) is involved in the key energy-producing pathway of glycolysis and presents a potential target for antibiotic development.AbPGK has been expressed and purified; it was crystallized using lithium sulfate as the precipitant. TheAbPGK crystals belonged to space groupP2221. They diffracted to a resolution of 2.5 Å using synchrotron radiation at the Canadian Light Source.

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Novel Engineered Peptides of a Phage Lysin as Effective Antimicrobials against Multidrug-Resistant Acinetobacter baumannii

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02972-15 ◽

2016 ◽

Vol 60 (5) ◽

pp. 2671-2679 ◽

Cited By ~ 52

Author(s):

Mya Thandar ◽

Rolf Lood ◽

Benjamin Y. Winer ◽

Douglas R. Deutsch ◽

Chad W. Euler ◽

...

Keyword(s):

Acinetobacter Baumannii ◽

Bacterial Pathogen ◽

Antibacterial Activities ◽

Polymyxin B ◽

Multidrug Resistant ◽

Gram Negative ◽

Content Type ◽

Human Red Blood Cells ◽

Phage Lysin

ABSTRACTAcinetobacter baumanniiis a Gram-negative bacterial pathogen responsible for a range of nosocomial infections. The recent rise and spread of multidrug-resistantA. baumanniiclones has fueled a search for alternative therapies, including bacteriophage endolysins with potent antibacterial activities. A common feature of these lysins is the presence of a highly positively charged C-terminal domain with a likely role in promoting outer membrane penetration. In the present study, we show that the C-terminal amino acids 108 to 138 of phage lysin PlyF307, named P307, alone were sufficient to killA. baumannii(>3 logs). Furthermore, P307 could be engineered for improved activity, the most active derivative being P307SQ-8C(>5-log kill). Both P307 and P307SQ-8Cshowed highin vitroactivity againstA. baumanniiin biofilms. Moreover, P307SQ-8Cexhibited MICs comparable to those of levofloxacin and ceftazidime and acted synergistically with polymyxin B. Although the peptides were shown to kill by disrupting the bacterial cytoplasmic membrane, they did not lyse human red blood cells or B cells; however, serum was found to be inhibitory to lytic activity. In a murine model ofA. baumanniiskin infection, P307SQ-8Creduced the bacterial burden by ∼2 logs in 2 h. This study demonstrates the prospect of using peptide derivatives from bacteriophage lysins to treat topical infections and remove biofilms caused by Gram-negative pathogens.

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MULTIDRUG-RESISTANT ACINETOBACTER BAUMANNII (MDRAB) PREVALENCE AND CHARACTERISTICS IN DR.WAHIDIN SUDIROHUSODO GENERAL HOSPITAL OF MAKASSAR

INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY ◽

10.24293/ijcpml.v25i2.1363 ◽

2019 ◽

Vol 25 (2) ◽

pp. 211

Author(s):

Dewi Tungadi ◽

Nurhayana Sennang ◽

Benny Rusli

Keyword(s):

Length Of Stay ◽

Acinetobacter Baumannii ◽

General Hospital ◽

Average Length ◽

Polymyxin B ◽

Multidrug Resistant ◽

Infection Prevention And Control ◽

Average Length Of Stay ◽

Improve Hospital ◽

And Control

BackgroundMultidrug-resistant Acinetobacter baumannii (MDRAB) is a strain of Acinetobacter baumannii which is resistant to three or more classes of antibiotics. As prevalence of MDRAB increases, the antibiotics of choice become limited. Identification of MDRAB is required to manage and control infection.MethodThis was a retrospective study, conducted in Dr. Wahidin Sudirohusodo General Hospital of Makassar, dated from January to December 2016. Bacterial identification and antimicrobial susceptibility testing (AST) were performed using VITEK 2. The patient data were obtained from electronic medical records.Results and DiscussionA total of 323 Acinetobacter baumannii isolates were obtained, consisted of 188 isolates in January-June 2016 and 36 of which was MDRAB (19.15%) with the average length-of-stay 33 days; and 135 isolates in July-December 2016 and 31 of which was MDRAB (22.96%) with the average length-of-stay 27 days. MDRAB was mostly discovered from patients using 3 or more medical devices and on single antibiotic therapy. MDRAB isolates were mostly obtained in sputum and pus specimens, and majority of patients had respiratory diseases. The result of AST showed 100% and 96% susceptibility to Polymyxin B; 71.43% and 54.84% susceptibility to Amikacin; 66.67% and 50% susceptibility to Trimethoprim/Sulfamethoxazole in January-June and July-December 2016, respectively.Conclusion and SuggestionsThe prevalence of MDRAB in our hospital in 2016 was high, suggesting the needs to improve hospital infection prevention and control. Polymyxin B, Amikacin, and Trimethoprim/Sulfamethoxazole are the antibiotics of choice to treat MDRAB.

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Molecular Dynamics Simulation-assisted Ionic Liquid Screening for Deep Coverage Proteome Analysis

Molecular & Cellular Proteomics ◽

10.1074/mcp.tir119.001827 ◽

2020 ◽

Vol 19 (10) ◽

pp. 1724-1737

Author(s):

Fei Fang ◽

Qun Zhao ◽

Huiying Chu ◽

Mingwei Liu ◽

Baofeng Zhao ◽

...

Keyword(s):

Molecular Dynamics ◽

Ionic Liquid ◽

Molecular Dynamics Simulation ◽

Membrane Proteins ◽

Sample Preparation ◽

Proteomic Analysis ◽

Dynamics Simulation ◽

Dominant Factor ◽

Label Free ◽

Compound Libraries

In-depth coverage of proteomic analysis could enhance our understanding to the mechanism of the protein functions. Unfortunately, many highly hydrophobic proteins and low-abundance proteins, which play critical roles in signaling networks, are easily lost during sample preparation, mainly attributed to the fact that very few extractants can simultaneously satisfy the requirements on strong solubilizing ability to membrane proteins and good enzyme compatibility. Thus, it is urgent to screen out ideal extractant from the huge compound libraries in a fast and effective way. Herein, by investigating the interior mechanism of extractants on the membrane proteins solubilization and trypsin compatibility, a molecular dynamics simulation system was established as complement to the experimental procedure to narrow down the scope of candidates for proteomics analysis. The simulation data shows that the van der Waals interaction between cation group of ionic liquid and membrane protein is the dominant factor in determining protein solubilization. In combination with the experimental data, 1-dodecyl-3-methylimidazolium chloride (C12Im-Cl) is on the shortlist for the suitable candidates from comprehensive aspects. Inspired by the advantages of C12Im-Cl, an ionic liquid-based filter-aided sample preparation (i-FASP) method was developed. Using this strategy, over 3,300 proteins were confidently identified from 103 HeLa cells (∼100 ng proteins) in a single run, an improvement of 53% over the conventional FASP method. Then the i-FASP method was further successfully applied to the label-free relative quantitation of human liver cancer and para-carcinoma tissues with obviously improved accuracy, reproducibility and coverage than the commonly used urea-based FASP method. The above results demonstrated that the i-FASP method could be performed as a versatile tool for the in-depth coverage proteomic analysis of biological samples.

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Origanum vulgare essential oil: antibacterial activities and synergistic effect with polymyxin B against multidrug-resistant Acinetobacter baumannii

Molecular Biology Reports ◽

10.1007/s11033-020-05989-0 ◽

2020 ◽

Vol 47 (12) ◽

pp. 9615-9625

Author(s):

Suélen Cavalheiro Amaral ◽

Beatriz Bohns Pruski ◽

Stella Buchhorn de Freitas ◽

Suzane Olachea Allend ◽

Marcos Roberto Alves Ferreira ◽

...

Keyword(s):

Essential Oil ◽

Synergistic Effect ◽

Acinetobacter Baumannii ◽

Antibacterial Activities ◽

Polymyxin B ◽

Multidrug Resistant ◽

Origanum Vulgare

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Polymyxin B and Doxycycline Use in Patients with Multidrug-Resistant Acinetobacter baumannii Infections in the Intensive Care Unit

Annals of Pharmacotherapy ◽

10.1345/aph.1h353 ◽

2006 ◽

Vol 40 (11) ◽

pp. 1939-1945 ◽

Cited By ~ 54

Author(s):

Katherine P Holloway ◽

Nadine G Rouphael ◽

Jane B Wells ◽

Mark D King ◽

Henry M Blumberg

Keyword(s):

Intensive Care Unit ◽

Intensive Care ◽

Acinetobacter Baumannii ◽

Polymyxin B ◽

Multidrug Resistant

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Phenotypic detection of extended-spectrum beta-lactamase in multidrug-resistant acinetobacter baumannii isolated in Fauji Foundation Hospital Rawalpindi

Journal of the Pakistan Medical Association ◽

10.47391/jpma.1265 ◽

2021 ◽

pp. 1-12

Author(s):

Asim Ali Shah ◽

Yasir Ali ◽

Ayesha Maqbool ◽

Shahid Ahmad Abbasi ◽

Admin

Keyword(s):

Acinetobacter Baumannii ◽

Sampling Technique ◽

Polymyxin B ◽

Multidrug Resistant ◽

Cross Sectional Study ◽

Beta Lactamase ◽

Cross Sectional ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Multiple Drugs

Abstract Objective: To evaluate the phenotypic detection of extended-spectrum beta-lactamase in multidrug-resistant acinetobacter baumannii. Methods: The cross-sectional study was conducted at the Department of Microbiology, Fauji Foundation Hospital, Rawalpindi, Pakistan, from August 2018 to April 2019, after the ethical approval from the Institutional Review Committee. Consecutive Non- probability sampling technique was used, and comprised clinical specimens, including pus, blood, sputum, urine, tracheal tubes and canula double lumen, which were processed using standard protocols. Colonies of acinetobacter baumannii were identified by gram staining and Analytical Profile Index-20E kit. Combination disc method was used for the identification of extended-spectrum beta-lactamse. Clinical and Laboratory Standards Institute guidelines were used for antimicrobial susceptibility. Data was analysed using SPSS 22 and Sample size was calculated by using earlier study with 5 % margin of error and 95 % confidence level. Results: Of the 78 isolates, 58(74.4%) related to females and 20(25.6%) to males. There was no extended-spectrum beta-lactamse producer. Imipenem, meropenem, cefotaxime, ampicillin and ceftazidime showed 100% resistance, while colistin and polymyxin B were sensitive to all strains. The incidence rate was high in samples isolated from tracheal tubes 47(60.3%), followed by pus 21(26.9%). Age was not found to be a significant factor (p>0.05). Conclusion: Acinetobacter baumannii showed a high resistance to multiple drugs and was not confined to any specific age group. Colistin and polymyxin B were found to be better choices. Continuous...

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