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9. 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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13. 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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14. Characterisation of a putative agr system in Clostridium botulinum and Clostridium sporogenes

Author

Cooksley, Clare Marie

Subjects
579, QR Microbiology
Abstract

Botulinum neurotoxin induces a potentially fatal paralytic condition in humans and various animal species collectively known as 'botulism'. It consequently poses a major problem to the food industry, due to the ability of its spores to survive in cooked foods. The incidence of wound botulism has also suffered a recent increase in the UK. The genome sequence of the C botulinum Group I strain ATCC 3502 has recently been determined. In silico analysis has revealed the presence of two distinct loci capable of encoding proteins with homology to AgrB and AgrD of the Staphylococcus aureus agr quorum sensing system. The functional characterisation of these genes has been carried out in order to determine whether they play a role in quorum sensing. To simplify laboratory procedures, C. sporogenes, the non-toxic counterpart of C. botulinum, was initially focused on. The agr regions in C. sporogenes were sequenced and their proteins compared with those of C. botulinum and other Gram-positive bacteria. Regions of conservation were observed amongst the clostridia and, to a lesser extent, between clostridia and staphylococci. Transcriptional linkage assays showed some of the genes of the C sporogenes agr regions to be co-expressed, and Real-Time RT-PCR demonstrated the maximal expression of these genes during late exponential growth. Modulation of the expression of the identified agr genes is a prerequisite to determining their function. Due to an initial lack of an effective gene knockout tool, antisense RNA expression was used for this purpose in C sporogenes, and showed that down regulation of the agrB genes affects sporulation. The development of an integrative vector system for gene inactivation in C sporogenes was also attempted. Knockout mutants in C botulinum and C sporogenes were later constructed using the newly-developed ClosTron system. These mutants were used to demonstrate that AgrDl, AgrD2 and an orphan sensor kinase protein all play a role in the control of sporulation in C. botulinum and C. sporogenes.

Published
2008

15. Chronic rhinosinusitis patients display an aberrant immune cell localization with enhanced S aureus biofilm metabolic activity and biomass

Author

Shaghayegh, Gohar, primary, Cooksley, Clare, additional, Bouras, George Spyro, additional, Panchatcharam, Beula Subashini, additional, Idrizi, Rejhan, additional, Jana, Metta, additional, Ellis, Sarah, additional, Psaltis, Alkis James, additional, Wormald, Peter-John, additional, and Vreugde, Sarah, 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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17. Wound healing in endoscopic sinus surgery: Phase 1 clinical trial evaluating the role of Chitogel with adjuvants

Author

Vediappan, Rajan Sundaresan, primary, Bennett, Catherine, additional, Cooksley, Clare, additional, Bassiouni, Ahmed, additional, Scott, John R., additional, Al Suliman, Yazeed A., additional, Lumyongsatien, Jate, additional, Moratti, Stephen, additional, Psaltis, Alkis J., additional, Vreugde, Sarah, additional, and Wormald, Peter‐John, additional

Published
2022
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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. Wound healing in endoscopic sinus surgery: Phase 1 clinical trial evaluating the role of Chitogel with adjuvants.

Author

Vediappan, Rajan Sundaresan, Bennett, Catherine, Cooksley, Clare, Bassiouni, Ahmed, Scott, John R., Al Suliman, Yazeed A., Lumyongsatien, Jate, Moratti, Stephen, Psaltis, Alkis J., Vreugde, Sarah, and Wormald, Peter‐John

Subjects
ENDOSCOPIC surgery, WOUND healing, CLINICAL trials, SPHENOID sinus, COMMERCIAL product testing, HEALING
Abstract

Objectives: This study aimed to determine the safety and efficacy of Chitogel, with and without Deferiprone (Def) and Gallium Protoporphyrin (GaPP), as a promoter of wound healing to improve surgical outcomes after endoscopic sinus susgery. Design: A double‐blinded, randomised control human clinical trial was conducted in patients undergoing ESS as a treatment for chronic rhinosinusitis. Participants underwent functional ESS or FESS with drill out as required and were randomised to receive test product Chitogel, Chitogel in combination with Def or Def‐GaPP versus no packing (control). Setting: Ostial stenosis and persistent inflammation are the main reasons for revision endoscopic sinus surgery (ESS). Post‐operative (PO) dressings can improve PO wound healing and patient outcomes after ESS. Participants: Eighty two patients were included in this study with 79 patients completing the study with 40 undergoing full house FESS and 39 FESS plus frontal drillout. Main Outcome Measures: Patients were followed up at 2, 6 and 12 weeks PO, and outcome scores such as SNOT‐22, VAS and LKS, pre and post‐surgery (12 weeks) were compared. Results: Seventy nine patients completed the study, there was a significant reduction in SNOT‐22 score and improvement of VAS at 12 weeks in patients treated with Chitogel compared to control (p <.05). In those patients, the mean ostium area for the Chitogel and the Chitogel + Def + GaPP groups was higher across all three sinuses compared to the no‐treatment control group, without statistical significance. Sphenoid sinus ostium was significantly more patent in patients treated with Chitogel compared to the control at the 12‐week time point (p <.05). Conclusion: Chitogel as a PO dressing after ESS results in the best patient‐reported symptom scores and objective measurements. The combination of Def and GaPP to Chitogel though proving safe, had no effect on the ostium patency or mucosal healing. [ABSTRACT FROM AUTHOR]

Published
2023
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34. Investigation of Kappa Carrageenan's muco‐adhesive, antibacterial, and anti‐biofilm properties.

Author

Javadiyan, Shari, Cooksley, Clare M, Bouras, George S., Kao, Stephen Shih‐Teng, Bennett, Catherine A., Wormald, Peter J., Vreugde, Sarah, and Psaltis, Alkis J.

Subjects
*CARRAGEENANS, *GRAM-positive bacteria, *BACTERIAL cell walls
Abstract

We have also shown, for the first time, that kappa carrageenan has antibacterial and anti-biofilm activity against I S. aureus i clinical isolates from the sinuses. I Staphylococcus aureus i has been reported to be the most common biofilm-forming pathogen in Chronic rhinosinusitis (CRS) pathogenesis and is associatedwith more severe disease and poor treatment response.1 There has been a recent increased interest in the use of topical antibiotic agents to treat CRS. The current study aimed to investigate both the muco-adhesiveness of kappa carrageenan on explanted sinonasal mucosa and its antibacterial and antibiofilm properties. [Extracted from the article]

Published
2022
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35. Prevention of adhesions post-abdominal surgery: Assessing the safety and efficacy of Chitogel with Deferiprone in a rat model

Author

Vediappan, Rajan Sundaresan, primary, Bennett, Catherine, additional, Cooksley, Clare, additional, Finnie, John, additional, Trochsler, Markus, additional, Quarrington, Ryan D., additional, Jones, Claire F., additional, Bassiouni, Ahmed, additional, Moratti, Stephen, additional, Psaltis, Alkis J., additional, Maddern, Guy, additional, Vreugde, Sarah, additional, and Wormald, P. J., additional

Published
2021
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37. Silver nanoparticles as a bioadjuvant of antibiotics against biofilm-mediated infections with methicillin-resistant Staphylococcus aureusand Pseudomonas aeruginosain chronic rhinosinusitis patients

Author

Feizi, Sholeh, Cooksley, Clare M., Nepal, Roshan, Psaltis, Alkis James, Wormald, Peter-John, and Vreugde, Sarah

Abstract

Infectious diseases caused by antibiotic-resistant bacteria in planktonic and biofilm form are difficult to treat with conventional antibiotics. Silver nanoparticles (Ag NPs) can be used as alternatives to antibiotics and can alter the susceptibility of bacteria to antibiotics. Here, the antibacterial properties of 16 different antibiotics and Ag NPs, alone and in combination, were tested against clinical isolates of Pseudomonas aeruginosa(n=3), Staphylococcus aureus(n=3) and methicillin-resistant Staphylococcus aureus(MRSA) (n=2) isolated from chronic rhinosinusitis (CRS) patients. The microdilution method and resazurin assay were used to determine the minimum inhibitory concentration and minimum biofilm eradication concentration for planktonic and biofilm forms, respectively. Results showed that Ag NPs and gentamicin combinations had synergistic antibacterial activity against P. aeruginosaplanktonic and biofilm forms and MRSA biofilms. Furthermore, additive effects against biofilms were seen for combinations of Ag NPs with tobramycin or ciprofloxacin against P. aeruginosa; with mupirocin against MRSA; and with augmentin, doxycycline, azithromycin and clindamycin against S. aureus. Moreover, additive effects against planktonic forms were observed for combinations of Ag NPs with tobramycin, ciprofloxacin, imipenem, ceftazidime and aztreonam against P. aeruginosa; with gentamicin or linezolid against MRSA; and with doxycycline or clindamycin against S. aureus. In conclusion, Ag NP-antibiotic combinations can result in enhanced antimicrobial action against P. aeruginosa,MRSA and S. aureusclinical isolates in planktonic and biofilm forms and can be used in the context of CRS with confirmed infection.

Published
2022
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38. ClosTron-Targeted Mutagenesis

Author

Heap, John T., primary, Cartman, Stephen T., additional, Kuehne, Sarah A., additional, Cooksley, Clare, additional, and Minton, Nigel P., additional

Published
2010
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41. 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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42. The international sinonasal microbiome study: A multicentre, multinational characterization of sinonasal bacterial ecology

Author

Paramasivan, Sathish, primary, Bassiouni, Ahmed, 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, 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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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, Ali, Mohammad Javed, additional, Bleier, Benjamin, 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
2019
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48. The international sinonasal microbiome study (ISMS): a multi-centre, multi-national collaboration characterising the microbial ecology of the sinonasal cavity

Author

Paramasivan, Sathish, primary, Bassiouni, Ahmed, additional, Shiffer, Arron, additional, Dillon, Matthew R, additional, Cope, Emily K, additional, Cooksley, Clare, additional, Ali, Mohammad Javed, additional, Bleier, Benjamin, 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
2019
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