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Item Comparison of Effectiveness of Germania Honey Compared to Manuka Honey in Methicillin-Resistant Staphylococcus aureus (MRSA) Killing(Bentham, 2019) Bazzi, Ali M.; Rabaan, Ali A.; Al-Tawfiq, Jaffar A.; Shannak, Bilal M.; Medicine, School of MedicinePurpose: Manuka honey is currently used in medical-grade sterile wound treatment products and has been shown to be effective in methicillin-resistant Staphylococcus aureus (MRSA) killing in vitro and in wound healing in a number of case studies and series. Locally produced honey in Pakistan and Chile have been proposed to be as effective as Manuka honey in bacterial killing in vitro, presenting potentially more accessible and affordable alternatives. In this study, we compared the effectiveness of a local Germania honey from Saudi Arabia to Manuka honey MGO 550 for in vitro killing of MRSA. Methodology: Overnight Muller Hinton broth cultures of 50 wound culture isolates of MRSA from 50 patients were incubated with a series of dilutions of Manuka honey MGO 550 and corresponding Germania honey dilutions for 24 h. Turbidity was assessed to determine whether bacterial growth had occurred, and no growth was confirmed by a further 24 h sub-culture on blood agar. Results/Key findings: Manuka honey MGO 550 was significantly more effective than Germania honey at MRSA killing at 100% v/v, 50% v/v and 25% v/v (p=0.025, 0.000265, and 0.000112 respectively) Conclusion: Manuka honey MGO 550 is significantly more effective in killing MRSA in vitro than Germania honey. Germania honey does not appear to be a promising locally produced alternative to Manuka honey for the development of honey-based wound dressings. Further experiments could determine if Germania honey is effective against other bacterial species.Item The Effect of Contact Precautions for MRSA on Patient Satisfaction Scores(Elsevier, 2015-07) Livorsi, Daniel J.; Kundu, Madan G.; Batteiger, Byron; Kressel, Amy B.; Department of Medicine, IU School of MedicineContact precautions may have an adverse effect on a patient's hospital experience and the delivery of care. This case–control study compared patient satisfaction scores between 70 patients isolated for MRSA and 139 non-isolated patients. Based on an adjusted analysis, there was no difference in patient satisfaction between the two groups. Age and educational status were found to affect patient satisfaction.Item Methicillin-Resistant Staphylococcus aureus (MRSA) Nasal Real-Time PCR: A Predictive Tool for Contamination of the Hospital Environment(Cambridge, 2015-01) Livorsi, Daniel J.; Arif, Sana; Garry, Patricia; Kundu, Madan G.; Satola, Sarah W.; Davis, Thomas H.; Batteiger, Byron; Kressel, Amy B.; Department of Medicine, IU School of MedicineOBJECTIVE We sought to determine whether the bacterial burden in the nares, as determined by the cycle threshold (CT) value from real-time MRSA PCR, is predictive of environmental contamination with MRSA. METHODS Patients identified as MRSA nasal carriers per hospital protocol were enrolled within 72 hours of room admission. Patients were excluded if (1) nasal mupirocin or chlorhexidine body wash was used within the past month or (2) an active MRSA infection was suspected. Four environmental sites, 6 body sites and a wound, if present, were cultured with premoistened swabs. All nasal swabs were submitted for both a quantitative culture and real-time PCR (Roche Lightcycler, Indianapolis, IN). RESULTS At study enrollment, 82 patients had a positive MRSA-PCR. A negative correlation of moderate strength was observed between the CT value and the number of MRSA colonies in the nares (r=−0.61; P<0.01). Current antibiotic use was associated with lower levels of MRSA nasal colonization (CT value, 30.2 vs 27.7; P<0.01). Patients with concomitant environmental contamination had a higher median log MRSA nares count (3.9 vs 2.5, P=0.01) and lower CT values (28.0 vs 30.2; P<0.01). However, a ROC curve was unable to identify a threshold MRSA nares count that reliably excluded environmental contamination. CONCLUSIONS Patients with a higher burden of MRSA in their nares, based on the CT value, were more likely to contaminate their environment with MRSA. However, contamination of the environment cannot be predicted solely by the degree of MRSA nasal colonization.Item Multihospital Infection Prevention Collaborative: Informatics Challenges and Strategies to Prevent MRSA(2013-11) Doebbeling, Bradley N.; Flanagan, Mindy E.; Nall, Glenna; Hoke, Shawn; Rosenman, Marc; Kho, AbelWe formed a collaborative to spread effective MRSA prevention strategies. We conducted a two-phase, multisite, quasi-experimental study of seven hospital systems (11 hospitals) in IN, MT, ME and Ontario, Canada over six years. Patients with prior MRSA were identified at admission using regional health information exchange data. We developed a system to return an alert message indicating a prior history of MRSA, directed to infection preventionists and admissions. Alerts indicated the prior anatomic site, and the originating institution. The combined approach of training and coaching, implementation of MRSA registries, notifying hospitals on admission of previously infected or colonized patients, and change strategies was effective in reducing MRSA infections over 80%. Further research and development of electronic surveillance tools is needed to better integrate the varied data source and support preventing MRSA infections. Our study supports the importance of hospitals collaborating to share data and implement effective strategies to prevent MRSA.Item A regional informatics platform for coordinated antibiotic resistant infection tracking, alerting and prevention(2013-04) Kho, Abel N.; Doebbeling, Bradley N.; Cashy, John P.; Rosenman, Marc B.; Dexter, Paul R.; Shepherd, David C.; Lemmon, Larry; Teal, Evgenia; Khokar, Shahid; Overhage, J. MarcBackground. We developed and assessed the impact of a patient registry and electronic admission notification system relating to regional antimicrobial resistance (AMR) on regional AMR infection rates over time. We conducted an observational cohort study of all patients identified as infected or colonized with methicillin-resistant Staphylococcus aureus (MRSA) and/or vancomycin-resistant enterococci (VRE) on at least 1 occasion by any of 5 healthcare systems between 2003 and 2010. The 5 healthcare systems included 17 hospitals and associated clinics in the Indianapolis, Indiana, region. Methods. We developed and standardized a registry of MRSA and VRE patients and created Web forms that infection preventionists (IPs) used to maintain the lists. We sent e-mail alerts to IPs whenever a patient previously infected or colonized with MRSA or VRE registered for admission to a study hospital from June 2007 through June 2010. Results. Over a 3-year period, we delivered 12 748 e-mail alerts on 6270 unique patients to 24 IPs covering 17 hospitals. One in 5 (22%–23%) of all admission alerts was based on data from a healthcare system that was different from the admitting hospital; a few hospitals accounted for most of this crossover among facilities and systems. Conclusions. Regional patient registries identify an important patient cohort with relevant prior antibiotic-resistant infection data from different healthcare institutions. Regional registries can identify trends and interinstitutional movement not otherwise apparent from single institution data. Importantly, electronic alerts can notify of the need to isolate early and to institute other measures to prevent transmission.Item Regulation of the Sae Two-Component System by Branched- Chain Fatty Acids in Staphylococcus aureus(American Society for Microbiology, 2022) Pendleton, Augustus; Yeo, Won-Sik; Alqahtani, Shahad; DiMaggio, Dennis A., Jr.; Stone, Carl J.; Li, Zhaotao; Singh, Vineet K.; Montgomery, Christopher P.; Bae, Taeok; Brinsmade, Shaun R.; Microbiology and Immunology, School of MedicineStaphylococcus aureus is a ubiquitous Gram-positive bacterium and an opportunistic human pathogen. S. aureus pathogenesis relies on a complex network of regulatory factors that adjust gene expression. Two important factors in this network are CodY, a repressor protein responsive to nutrient availability, and the SaeRS two-component system (TCS), which responds to neutrophil-produced factors. Our previous work revealed that CodY regulates the secretion of many toxins indirectly via Sae through an unknown mechanism. We report that disruption of codY results in increased levels of phosphorylated SaeR (SaeR~P) and that codY mutant cell membranes contain a higher percentage of branched-chain fatty acids (BCFAs) than do wild-type membranes, prompting us to hypothesize that changes to membrane composition modulate the activity of the SaeS sensor kinase. Disrupting the lpdA gene encoding dihydrolipoyl dehydrogenase, which is critical for BCFA synthesis, significantly reduced the abundance of SaeR, phosphorylated SaeR, and BCFAs in the membrane, resulting in reduced toxin production and attenuated virulence. Lower SaeR levels could be explained in part by reduced stability. Sae activity in the lpdA mutant could be complemented genetically and chemically with exogenous short- or full-length BCFAs. Intriguingly, lack of lpdA also alters the activity of other TCSs, suggesting a specific BCFA requirement managing the basal activity of multiple TCSs. These results reveal a novel method of posttranscriptional virulence regulation via BCFA synthesis, potentially linking CodY activity to multiple virulence regulators in S. aureus.Item Role of p85α in neutrophil extra- and intracellular reactive oxygen species generation(Impact Journals, 2016-04-26) Li, Xing Jun; Deng, Lisa; Brandt, Stephanie L.; Goodwin, Charles B.; Ma, Peilin; Yang, Zhenyun; Mali, Raghu S.; Liu, Ziyue; Kapur, Reuben; Serezani, C. Henrique; Chan, Rebecca J.; Department of Pediatrics, IU School of MedicineDrug resistance is a growing problem that necessitates new strategies to combat pathogens. Neutrophil phagocytosis and production of intracellular ROS, in particular, has been shown to cooperate with antibiotics in the killing of microbes. This study tested the hypothesis that p85α, the regulatory subunit of PI3K, regulates production of intracellular ROS. Genetic knockout of p85α in mice caused decreased expression of catalytic subunits p110α, p110β, and p110δ, but did not change expression levels of the NADPH oxidase complex subunits p67phox, p47phox, and p40phox. When p85α, p55α, and p50α (all encoded by Pik3r1) were deleted, there was an increase in intracellular ROS with no change in phagocytosis in response to both Fcγ receptor and complement receptor stimulation. Furthermore, the increased intracellular ROS correlated with significantly improved neutrophil killing of both methicillin-susceptible and methicillin-resistant S. aureus. Our findings suggest inhibition of p85α as novel approach to improving the clearance of resistant pathogens.Item Unravelling the physiological roles of mazEF toxin–antitoxin system on clinical MRSA strain by CRISPR RNA-guided cytidine deaminase(Springer Nature, 2022-05-07) Jain, Sonia; Bhowmick, Arghya; Jeong, Bohyun; Bae, Taeok; Ghosh, Abhrajyoti; Microbiology and Immunology, School of MedicineBackground: Curiosity on toxin-antitoxin modules has increased intensely over recent years as it is ubiquitously present in many bacterial genomes, including pathogens like Methicillin-resistant Staphylococcus aureus (MRSA). Several cellular functions of TA systems have been proposed however, their exact role in cellular physiology remains unresolved. Methods: This study aims to find out the impact of the mazEF toxin-antitoxin module on biofilm formation, pathogenesis, and antibiotic resistance in an isolated clinical ST239 MRSA strain, by constructing mazE and mazF mutants using CRISPR-cas9 base-editing plasmid (pnCasSA-BEC). Transcriptome analysis (RNA-seq) was performed for the mazE antitoxin mutant in order to identify the differentially regulated genes. The biofilm formation was also assessed for the mutant strains. Antibiogram profiling was carried out for both the generated mutants followed by murine experiment to determine the pathogenicity of the constructed strains. Results: For the first time our work showed, that MazF promotes cidA mediated cell death and lysis for biofilm formation without playing any significant role in host virulence as suggested by the murine experiment. Interestingly, the susceptibility to oxacillin, daptomycin and vancomycin was reduced significantly by the activated MazF toxin in the mazE mutant strain. Conclusions: Our study reveals that activated MazF toxin leads to resistance to antibiotics like oxacillin, daptomycin and vancomycin. Therefore, in the future, any potential antibacterial drug can be designed to target MazF toxin against the problematic multi-drug resistant bug.