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6. Increased antibiotic resistance of Pseudomonas aeruginosa isolates from chronic rhinosinusitis patients grown in anaerobic conditions.

Author

Ferdoush, Jannatul, Ramezanpour, Mahnaz, Cooksley, Clare, Bouras, George Spyro, Kazuhiro Ogi, Feizi, Sholeh, Nepal, Roshan, Psaltis, Alkis James, Wormald, Peter-John, and Vreugde, Sarah

Subjects
DRUG resistance in bacteria, PSEUDOMONAS aeruginosa, SINUSITIS, ANAEROBIC bacteria, ANAEROBIC microorganisms, DRUG resistance in microorganisms
Abstract

Introduction: In chronic rhinosinusitis (CRS), the congestion and blockage of the nose can cause anaerobic conditions within the sinus cavities which may promote the expression of virulence and antibiotic resistance genes in invading pathogens. Pseudomonas aeruginosa is a facultative anaerobic bacteria and causes severe recalcitrant CRS. In this study, we aimed to evaluate the antimicrobial resistance of P. aeruginosa isolates of CRS patients in planktonic and biofilm form grown in aerobic and anaerobic conditions. Methods: P. aeruginosa clinical isolates of CRS patients (n = 25) were grown in planktonic and biofilm form in aerobic and anaerobic conditions. Minimum inhibitory concentrations (MIC) of planktonic forms and minimum biofilm eradication concentrations (MBEC) were determined. Additionally, metabolic activity by fluorescein diacetate assay, biofilm biomass by crystal violet assay and eDNA concentration were assessed in both conditions. Results: P. aeruginosa planktonic cells grown in anaerobic condition exhibited increased gentamicin resistance (p < .01), whereas P. aeruginosa biofilms grown in anaerobic condition displayed significantly increased MBEC values for gentamicin (p < .0001) and levofloxacin (p < .001). The metabolic activity of anaerobic biofilms was significantly higher compared with aerobic biofilms (p < .0001). However, the biofilm biomass of isolates grown in aerobic conditions was higher than anaerobic conditions (p < .5). Conclusion: P. aeruginosa isolates from CRS patients grown in anaerobic conditions showed significantly increased resistance to antibiotics with an increased metabolic activity but decreased biofilm biomass. [ABSTRACT FROM AUTHOR]

Published
2024
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12. Staphylococcus aureus biofilm properties and chronic rhinosinusitis severity scores correlate positively with total CD4 + T‐ cell frequencies and inversely with its Th1, Th17 and regulatory cell frequencies

Author

Shaghayegh, Gohar, primary, Cooksley, Clare, additional, Bouras, George, additional, Nepal, Roshan, additional, Houtak, Ghais, additional, Panchatcharam, Beula Subashini, additional, Fenix, Kevin Aaron, additional, Psaltis, Alkis James, additional, Wormald, Peter‐John, additional, and Vreugde, Sarah, additional

Published
2023
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15. Numerical and Machine Learning Analysis of the Parameters Affecting the Regionally Delivered Nasal Dose of Nano- and Micro-Sized Aerosolized Drugs

Author

Farnoud, Ali, primary, Tofighian, Hesam, additional, Baumann, Ingo, additional, Ahookhosh, Kaveh, additional, Pourmehran, Oveis, additional, Cui, Xinguang, additional, Heuveline, Vincent, additional, Song, Chen, additional, Vreugde, Sarah, additional, Wormald, Peter-John, additional, Menden, Michael P., additional, and Schmid, Otmar, additional

Published
2023
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16. Staphylococcus aureus biofilm properties and chronic rhinosinusitis severity scores correlate positively with total CD4+ T‐cell frequencies and inversely with its Th1, Th17 and regulatory cell frequencies.

Author

Shaghayegh, Gohar, Cooksley, Clare, Bouras, George, Nepal, Roshan, Houtak, Ghais, Panchatcharam, Beula Subashini, Fenix, Kevin Aaron, Psaltis, Alkis James, Wormald, Peter‐John, and Vreugde, Sarah

Subjects
ENDOSCOPIC surgery, NASAL polyps, T helper cells, T cells, BACTERIAL colonies, CD4 antigen, STAPHYLOCOCCUS aureus
Abstract

Chronic rhinosinusitis (CRS) represents chronic inflammation of the sinus mucosa characterised by dysfunction of the sinuses' natural defence mechanisms and induction of different inflammatory pathways ranging from a Th1 to a Th2 predominant polarisation. Recalcitrant CRS is associated with Staphylococcus aureus dominant mucosal biofilms; however, S. aureus colonisation of the sinonasal mucosa has also been observed in healthy individuals challenging the significance of S. aureus in CRS pathogenesis. We aimed to investigate the relationship between CRS key inflammatory markers, S. aureus biofilm properties/virulence genes and the severity of the disease. Tissue samples were collected during endoscopic sinus surgery from the ethmoid sinuses of CRS patients with (CRSwNP) and without (CRSsNP) nasal polyps and controls (n = 59). CD3+ T‐cell subset frequencies and key inflammatory markers of CD4+ helper T cells were determined using FACS analysis. Sinonasal S. aureus clinical isolates were isolated (n = 26), sequenced and grown into biofilm in vitro, followed by determining their properties, including metabolic activity, biomass, colony‐forming units and exoprotein production. Disease severity was assessed using Lund–Mackay radiologic scores, Lund–Kennedy endoscopic scores and SNOT22 quality of life scores. Our results showed that S. aureus biofilm properties and CRS severity scores correlated positively with total CD4+ T‐cell frequencies but looking into CD4+ T‐cell subsets showed an inverse correlation with Th1 and Th17 cell frequencies. CD4+ T‐cell frequencies were higher in patients harbouring lukF.PV‐positive S. aureus while its regulatory and Th17 cell subset frequencies were lower in patients carrying sea− and sarT/U‐positive S. aureus. Recalcitrant CRS is characterised by increased S. aureus biofilm properties in relation to increased total CD4+ helper T‐cell frequencies and reduced frequencies of its Th1, Th17 and regulatory T‐cell subsets. These findings offer insights into the pathophysiology of CRS and could lead to the development of more targeted therapies. [ABSTRACT FROM AUTHOR]

Published
2023
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19. Metallothionein-3 is a clinical biomarker for tissue zinc levels in nasal mucosa

Author

Suzuki, Masanobu, Ramezanpour, Mahnaz, Cooksley, Clare, Ogi, Kazuhiro, Psaltis, Alkis J., Nakamaru, Yuji, Homma, Akihiro, Wormald, Peter-John, and Vreugde, Sarah

Subjects
And nasal polyps, Chronic rhinosinusitis, Zinquin, Human nasal epithelial cells
Abstract

Objective: Recently, depleted tissue zinc levels were found in nasal mucosa from patients with chronic rhinosinusitis (CRS) in correlation with tissue eosinophilia, however, no clinical biomarkers for tissue zinc levels have been identified. Metallothionein-3 (MT3) is an intracellular zinc chelator and previous data showed MT3 mRNA levels to be reduced in CRS patients with nasal polyps (CRSwNP). In this study, we examined the correlation between MT3 expression and zinc levels in nasal mucosa and primary human nasal epithelial cells (HNECs) to investigate whether MT3 could be a clinical biomarker for tissue zinc levels. Method: Tissue was harvested from 36 patients and mounted on tissue micro-array (TMA) slides. MT3 expression and tissue zinc fluorescence intensity were measured at different areas within the mucosa (surface epithelium and lamina propria) and compared between controls, CRSwNP and CRS without nasal polyps (CRSsNP) patients. MT3 mRNA and protein expression were examined in zinc-depleted HNECs by qPCR and immunofluorescence microscopy. Results: MT3 expression in CRSwNP was significantly decreased in both surface epithelium (p < 0.001 to controls) and lamina propria (p = 0.0491 to controls). There was a significant positive correlation between tissue zinc levels and MT3 expression in nasal mucosa (r = 0.45, p = 0.007). In zinc-deplete HNECs, MT3 expression was significantly decreased at mRNA (p = 0.02) and protein level (p < 0.01). There was a significant positive correlation between tissue zinc levels and MT3 expression within individual HNECs (r = 0.59, p < 0.001). Conclusions: MT3 expression reflects intramucosal zinc levels in both nasal mucosa and HNECs indicating MT3 could be used as a clinical biomarker for monitoring intracellular zinc levels in the nasal mucosa. (C) 2021 Oto-Rhino-Laryngological Society of Japan Inc. Published by Elsevier B.V. All rights reserved.

Published
2021

22. Repetitive simulation training with novel 3D ‐printed sinus models for functional endoscopic sinus surgeries

Author

Suzuki, Masanobu, primary, Miyaji, Kou, additional, Watanabe, Ryosuke, additional, Suzuki, Takayoshi, additional, Matoba, Kotaro, additional, Nakazono, Akira, additional, Nakamaru, Yuji, additional, Konno, Atsushi, additional, Psaltis, Alkis James, additional, Abe, Takashige, additional, Homma, Akihiro, additional, and Wormald, Peter‐John, additional

Published
2022
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24. Efficacy and Safety of Novel Beta-Chitin Patches as Haemostat in Rat Vascular and Neurosurgical Model

Author

Sabab, Ahad, Vediappan, Rajan Sundaresan, Finnie, John, McAdam, C. John, Jukes, Alistair, Vreugde, Sarah, and Wormald, Peter-John

Subjects
Surgery
Abstract

BackgroundIntraoperative hemorrhage is a major cause of poor post-operative outcome. Beta-chitin patch has previously been found to be an effective haemostat, but whether modifying the patch can improve its efficacy and safety, remains unknown. In this study, beta-chitin patches were modified using polyethylene oxide, Pluronic-F127 (Chi/F127), calcium (Chi/20%Ca), increased thickness (Chi/Thick) or polyphosphate (Chi/PP).ObjectiveUsing rat (Wistar Albino; 8–10 weeks old) vascular and neurosurgical models, this project investigated and compared the efficacy and safety of beta-chitin patches with gauze, Surgicel and FloSeal.MethodsNinety rats underwent a standardized femoral artery injury and were randomized to receive either beta-chitin patches, gauze, Surgicel or FloSeal. The bleeding time and total blood loss was measured. For the neurosurgical model, forty-four rats underwent a standardized cortical injury and randomization to a treatment group. Following a 48 h recovery period, their brains were collected for histopathological examination.ResultsThe mean bleeding time with Chitin (120.8 s) and Chi/PP (117.3 s) was ~60 s lower than Chi/F127, Chi/20%Ca and Chi/Thick (p < 0.05). Chitin and Chi/PP had a significantly lower bleeding time than FloSeal (174.2 s) (p < 0.05), but not Surgicel (172.7 s). Gauze (400 s) had a significantly higher bleeding time compared to all other groups (p < 0.05). There were no significant differences in the total blood loss between the groups. Histopathological examination of brains found no adverse inflammatory reaction to any of the haemostatic compounds.ConclusionChi/PP had superior haemostatic efficacy compared to Surgicel and FloSeal, but not compared to non-modified beta-chitin patch. All of the haemostats were equally safe.

Published
2022

29. Pharokka: a fast scalable bacteriophage annotation tool.

Author

Bouras, George, Nepal, Roshan, Houtak, Ghais, Psaltis, Alkis James, Wormald, Peter-John, and Vreugde, Sarah

Subjects
VIRTUAL machine systems, BACTERIOPHAGES, SOURCE code, PRODUCTION standards, ANNOTATIONS
Abstract

Summary In recent years, there has been an increasing interest in bacteriophages, which has led to growing numbers of bacteriophage genomic sequences becoming available. Consequently, there is a need for a rapid and consistent genomic annotation tool dedicated for bacteriophages. Existing tools either are not designed specifically for bacteriophages or are web- and email-based and require significant manual curation, which makes their integration into bioinformatic pipelines challenging. Pharokka was created to provide a tool that annotates bacteriophage genomes easily, rapidly and consistently with standards compliant outputs. Moreover, Pharokka requires only two lines of code to install and use and takes under 5 min to run for an average 50-kb bacteriophage genome. Availability and implementation Pharokka is implemented in Python and is available as a bioconda package using 'conda install -c bioconda pharokka'. The source code is available on GitHub (https://github.com/gbouras13/pharokka). Pharokka has been tested on Linux-64 and MacOSX machines and on Windows using a Linux Virtual Machine. [ABSTRACT FROM AUTHOR]

Published
2023
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39. International Consensus Statement on Rhinology and Allergy: Rhinosinusitis

Author

Orlandi, Richard, Kingdom, Todd T., Smith, Timothy L., Bleier, Benjamin, DeConde, Adam, Luong, Amber, Poetker, David M., Soler, Zachary, Welch, Kevin C., Wise, Sarah K., Adappa, Nithin, Alt, Jeremiah A., Anselmo-Lima, Wilma Terezinha, Bachert, Claus, Baroody, Fuad M., Batra, Pete S., Bernal-Sprekelsen, Manuel, Beswick, Daniel, Bhattacharyya, Neil, Chandra, Rakesh K., Chang, Eugene, Chiu, Alexander, Chowdhury, Naweed, Citardi, Martin J., Cohen, Noam, Conley, David B., DelGaudio, John, Desrosiers, Martin, Douglas, Richard, Anderson Eloy, Jean, Fokkens, Wytske J., Gray, Stacey T., Gudis, David A., Hamilos, Daniel L., Han, Joseph K., Harvey, Richard, Hellings, Peter, Holbrook, Eric H., Hopkins, Claire, Hwang, Peter, Javer, Amin R., Jiang, Rong-San, Kennedy, David, Kern, Robert, Laidlaw, Tanya, Lal, Devyani, Lane, Andrew, Lee, Heung-Man, Lee, Jivianne T., Levy, Joshua M., Lin, Sandra Y., Lund, Valerie, McMains, Kevin C., Metson, Ralph, Mullol, Joaquim, Naclerio, Robert, Oakley, Gretchen, Otori, Nobuyoshi, Palmer, James N., Parikh, Sanjay R., Passali, Desiderio, Patel, Zara, Peters, Anju, Philpott, Carl, Psaltis, Alkis J., Ramakrishnan, Vijay R., Ramanathan, Murugappan, Roh, Hwan-Jung, Rudmik, Luke, Sacks, Raymond, Schlosser, Rodney J., Sedaghat, Ahmad, Senior, Brent, Sindwani, Raj, Smith, Kristine, Snidvongs, Kornkiat, Stewart, Michael, Suh, Jeffrey, Tan, Bruce K., Turner, Justin H., van Drunen, Cornelis M., Voegels, Richard, Wang, De-Yun, Woodworth, Bradford A., Wormald, Peter-John, Wright, Erin D., Yan, Carol, Zhang, Luo, and Zhou, Bing

Abstract

Background: The 5 years since the publication of the first International Consensus Statement on Allergy and Rhinology: Rhinosinusitis (ICAR‐RS) has witnessed foundational progress in our understanding and treatment of rhinologic disease. These advances are reflected within the more than 40 new topics covered within the ICAR‐RS‐2021 as well as updates to the original 140 topics. This executive summary consolidates the evidence‐based findings of the document. Methods: ICAR‐RS presents over 180 topics in the forms of evidence‐based reviews with recommendations (EBRRs), evidence‐based reviews, and literature reviews. The highest grade structured recommendations of the EBRR sections are summarized in this executive summary. Results: ICAR‐RS‐2021 covers 22 topics regarding the medical management of RS, which are grade A/B and are presented in the executive summary. Additionally, 4 topics regarding the surgical management of RS are grade A/B and are presented in the executive summary. Finally, a comprehensive evidence‐based management algorithm is provided. Conclusion: This ICAR‐RS‐2021 executive summary provides a compilation of the evidence‐based recommendations for medical and surgical treatment of the most common forms of RS.

Published
2021

40. Corynebacterium accolens inhibits Staphylococcus aureus induced mucosal barrier disruption.

Author

Shuman Huang, Hon, Karen, Bennett, Catherine, Hua Hu, Menberu, Martha, Wormald, Peter-John, Yulin Zhao, Vreugde, Sarah, and Sha Liu

Subjects
STAPHYLOCOCCUS aureus, CORYNEBACTERIUM, LACTATE dehydrogenase, LASER microscopy, EPITHELIAL cells
Abstract

Background: Corynebacterium accolens (C. accolens) is a common nasal colonizer, whereas Staphylococcus aureus (S. aureus) is typically regarded a pathogenic organism in patients with chronic rhinosinusitis (CRS). This study aims to evaluate the interaction of the two bacteria in vitro. Methods: Clinical isolates of C. accolens and S. aureus from sinonasal swabs, as well as primary human nasal epithelial cells (HNECs) cultured from cellular brushings of both healthy and CRS patients were used for this study. The cell-free culture supernatants of all isolates grown alone and in co-cultures were tested for their effects on transepithelial electrical resistance (TER), FITCDextran permeability, lactate dehydrogenase (LDH), and IL-6 and IL-8 secretion of HNECs. Confocal scanning laser microscopy and immunofluorescence were also used to visualize the apical junctional complexes. C. accolens cell-free culture supernatants were also tested for antimicrobial activity and growth on planktonic and biofilm S. aureus growth. Results: The cell-free culture supernatants of 3\C. accolens strains (at 60% for S. aureus reference strain and 30% concentration for S. aureus clinical strains) inhibited the growth of both the planktonic S. aureus reference and clinical strains significantly. The C. accolens cell-free culture supernatants caused no change in the TER or FITC-Dextran permeability of the HNEC-ALI cultures, while the cell-free culture supernatants of S. aureus strains had a detrimental effect. Cell-free culture supernatants of C. accolens co-cultured with both the clinical and reference strains of S. aureus delayed the S. aureus-dependent mucosal barrier damage in a dose-dependent manner. Conclusion: Corynebacterium accolens cell-free culture supernatants appear to inhibit the growth of the S. aureus planktonic bacteria, and may reduce the mucosal barrier damage caused by S. aureus. [ABSTRACT FROM AUTHOR]

Published
2022
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47. Microbiotyping the Sinonasal Microbiome

Author

Bassiouni, Ahmed, primary, Paramasivan, Sathish, additional, Shiffer, Arron, additional, Dillon, Matthew R., additional, Cope, Emily K., additional, Cooksley, Clare, additional, Ramezanpour, Mahnaz, additional, Moraitis, Sophia, additional, Ali, Mohammad Javed, additional, Bleier, Benjamin S., additional, Callejas, Claudio, additional, Cornet, Marjolein E., additional, Douglas, Richard G., additional, Dutra, Daniel, additional, Georgalas, Christos, additional, Harvey, Richard J., additional, Hwang, Peter H., additional, Luong, Amber U., additional, Schlosser, Rodney J., additional, Tantilipikorn, Pongsakorn, additional, Tewfik, Marc A., additional, Vreugde, Sarah, additional, Wormald, Peter-John, additional, Caporaso, J. Gregory, additional, and Psaltis, Alkis J., additional

Published
2020
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