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2. Paeniclostridium (Clostridium) sordellii–associated enterocolitis in 7 horses

Author

Nyaoke, Akinyi C, Navarro, Mauricio A, Fresneda, Karina, Diab, Santiago S, Moore, Janet, Lyras, Dena, Awad, Milena, and Uzal, Francisco A

Subjects
Digestive Diseases, Emerging Infectious Diseases, Good Health and Well Being, Animals, Clostridium, Clostridium Infections, Clostridium sordellii, Diagnosis, Differential, Enterocolitis, Horse Diseases, Horses, Intestine, Large, Intestine, Small, colitis, enteritis, enterocolitis, horses, Paeniclostridium sordellii, Zoology, Veterinary Sciences
Abstract

Enteric disease in horses may be caused by a variety of microorganisms, including several clostridial species. Paeniclostridium sordellii (previously Clostridium sordellii) has been frequently associated with gas gangrene in humans and several animal species, including horses. However, its role in enteric diseases of animals has not been fully determined. We describe herein 7 cases of enteric disease in horses associated with P. sordellii infection. Grossly, the small and/or large intestines were necrotic, hemorrhagic, and edematous. Microscopically, there was severe mucosal necrosis and hemorrhage of the small and/or large intestine of all horses. P. sordellii was isolated and/or demonstrated by immunohistochemistry and/or PCR in the intestine of all horses. All other known causes of enteric disease in horses were ruled out in these 7 cases. P. sordellii should be considered among the differential diagnoses in cases of enteric disease in horses.

Published
2020

4. Towards an understanding of the role of Clostridium perfringens toxins in human and animal disease

Author

Uzal, Francisco A, Freedman, John C, Shrestha, Archana, Theoret, James R, Garcia, Jorge, Awad, Milena M, Adams, Vicki, Moore, Robert J, Rood, Julian I, and McClane, Bruce A

Subjects
Microbiology, Biological Sciences, Foodborne Illness, Emerging Infectious Diseases, Infectious Diseases, Digestive Diseases, 2.2 Factors relating to the physical environment, Aetiology, Infection, Animal Diseases, Animals, Bacterial Toxins, Clostridium Infections, Clostridium perfringens, Gas Gangrene, Humans, Intestines, animal disease, avian necrotic enteritis, enterocolitis, enterotoxemia, food poisoning, gas gangrene, human disease, toxins, Medical Microbiology
Abstract

Clostridium perfringens uses its arsenal of >16 toxins to cause histotoxic and intestinal infections in humans and animals. It has been unclear why this bacterium produces so many different toxins, especially since many target the plasma membrane of host cells. However, it is now established that C. perfringens uses chromosomally encoded alpha toxin (a phospholipase C) and perfringolysin O (a pore-forming toxin) during histotoxic infections. In contrast, this bacterium causes intestinal disease by employing toxins encoded by mobile genetic elements, including C. perfringens enterotoxin, necrotic enteritis toxin B-like, epsilon toxin and beta toxin. Like perfringolysin O, the toxins with established roles in intestinal disease form membrane pores. However, the intestinal disease-associated toxins vary in their target specificity, when they are produced (sporulation vs vegetative growth), and in their sensitivity to intestinal proteases. Producing many toxins with diverse characteristics likely imparts virulence flexibility to C. perfringens so it can cause an array of diseases.

Published
2014

5. Clostridium perfringens virulence factors are nonredundant activators of the NLRP3 inflammasome

Author

Mathur, Anukriti, primary, Kay, Callum, additional, Xue, Yansong, additional, Pandey, Abhimanu, additional, Lee, Jiwon, additional, Jing, Weidong, additional, Enosi Tuipulotu, Daniel, additional, Lo Pilato, Jordan, additional, Feng, Shouya, additional, Ngo, Chinh, additional, Zhao, Anyang, additional, Shen, Cheng, additional, Rug, Melanie, additional, Miosge, Lisa A, additional, Atmosukarto, Ines I, additional, Price, Jason D, additional, Ali, Sidra A, additional, Gardiner, Elizabeth E, additional, Robertson, Avril AB, additional, Awad, Milena M, additional, Lyras, Dena, additional, Kaakoush, Nadeem O, additional, and Man, Si Ming, additional

Published
2023
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13. Clostridium septicum α-toxin activates the NLRP3 inflammasome by engaging GPI-anchored proteins

Author

Jing, Weidong, primary, Pilato, Jordan Lo, additional, Kay, Callum, additional, Feng, Shouya, additional, Tuipulotu, Daniel Enosi, additional, Mathur, Anukriti, additional, Shen, Cheng, additional, Ngo, Chinh, additional, Zhao, Anyang, additional, Miosge, Lisa A., additional, Ali, Sidra A., additional, Gardiner, Elizabeth E., additional, Awad, Milena M., additional, Lyras, Dena, additional, Robertson, Avril A. B., additional, Kaakoush, Nadeem O., additional, and Man, Si Ming, additional

Published
2022
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15. PGFinder, a novel analysis pipeline for the consistent, reproducible, and high-resolution structural analysis of bacterial peptidoglycans

Author

Patel, Ankur V, primary, Turner, Robert D, additional, Rifflet, Aline, additional, Acosta-Martin, Adelina E, additional, Nichols, Andrew, additional, Awad, Milena M, additional, Lyras, Dena, additional, Gomperts Boneca, Ivo, additional, Bern, Marshall, additional, Collins, Mark O, additional, and Mesnage, Stéphane, additional

Published
2021
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16. Author response: PGFinder, a novel analysis pipeline for the consistent, reproducible, and high-resolution structural analysis of bacterial peptidoglycans

Author

Patel, Ankur V, primary, Turner, Robert D, additional, Rifflet, Aline, additional, Acosta-Martin, Adelina E, additional, Nichols, Andrew, additional, Awad, Milena M, additional, Lyras, Dena, additional, Gomperts Boneca, Ivo, additional, Bern, Marshall, additional, Collins, Mark O, additional, and Mesnage, Stéphane, additional

Published
2021
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17. PGfinder, a novel analysis pipeline for the consistent, reproducible and high-resolution structural analysis of bacterial peptidoglycans

Author

Patel, Ankur V., primary, Turner, Robert D., additional, Rifflet, Aline, additional, Acosta-Martin, Adelina E., additional, Nichols, Andrew, additional, Awad, Milena M., additional, Lyras, Dena, additional, Gomperts-Boneca, Ivo, additional, Bern, Marshall, additional, Collins, Mark O., additional, and Mesnage, Stéphane, additional

Published
2021
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18. Role of Clostridium perfringens toxins in the pathogenesis of clostridial myonecrosis

Author

Awad., Milena Mary

Subjects
ComputingMilieux_COMPUTERSANDEDUCATION, ComputerApplications_COMPUTERSINOTHERSYSTEMS, Uncategorized
Abstract

This thesis was scanned from the print manuscript for digital preservation and is copyright the author. Researchers can access this thesis by asking their local university, institution or public library to make a request on their behalf. Monash staff and postgraduate students can use the link in the References field.

Published
2021
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20. A dynamic, ring-forming MucB / RseB-like protein influences spore shape in Bacillus subtilis

Author

Kearns, Daniel B., Luhur, Johana, Chan, Helena, Kachappilly, Benson, Mohamed, Ahmed, Morlot, Cécile, Awad, Milena, Lyras, Dena, Taib, Najwa, Gribaldo, Simonetta, Rudner, David Z., Rodrigues, Christopher D.A., University of Technology Sydney (UTS), Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Department of Microbiology [Monash University, Australia], School of Biomedical Sciences [Monash University, Clayton], Monash University [Clayton]-Monash University [Clayton], Biologie Evolutive de la Cellule Microbienne - Evolutionary Biology of the Microbial Cell, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Harvard Medical School [Boston] (HMS), This work was funded by grant DP190100793 awarded to C.D.A.R from the Australian Research Council (https://www.arc.gov.au), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Cancer Research, Mutant, Cell Membranes, Bacillus subtilis, QH426-470, Endospore, MESH: Spores, Bacterial, Cell Wall, Microbial Physiology, Fluorescence microscope, Bacterial Physiology, Cell Cycle and Cell Division, MESH: Bacterial Proteins, Genetics (clinical), Spores, Bacterial, Staining, 0303 health sciences, Bacterial Sporulation, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Cell biology, Membrane Staining, Aspect Ratio, Cell Processes, Physical Sciences, MESH: Protein Domains, Cell envelope, Cellular Structures and Organelles, Research Article, MESH: Mutation, Imaging Techniques, Protein domain, Geometry, Biology, Research and Analysis Methods, Microbiology, 03 medical and health sciences, MESH: Cell Wall, Bacterial Proteins, Protein Domains, Fluorescence Imaging, Genetics, Compartment (development), Bacterial Spores, Molecular Biology, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, 030306 microbiology, QH, fungi, Biology and Life Sciences, Membrane Proteins, Bacteriology, Cell Biology, MESH: Bacillus subtilis, biology.organism_classification, Outer Membrane Proteins, QP, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, QR, Spore, Specimen Preparation and Treatment, Mutation, Mathematics
Abstract

How organisms develop into specific shapes is a central question in biology. The maintenance of bacterial shape is connected to the assembly and remodelling of the cell envelope. In endospore-forming bacteria, the pre-spore compartment (the forespore) undergoes morphological changes that result in a spore of defined shape, with a complex, multi-layered cell envelope. However, the mechanisms that govern spore shape remain poorly understood. Here, using a combination of fluorescence microscopy, quantitative image analysis, molecular genetics and transmission electron microscopy, we show that SsdC (formerly YdcC), a poorly-characterized new member of the MucB / RseB family of proteins that bind lipopolysaccharide in diderm bacteria, influences spore shape in the monoderm Bacillus subtilis. Sporulating cells lacking SsdC fail to adopt the typical oblong shape of wild-type forespores and are instead rounder. 2D and 3D-fluorescence microscopy suggest that SsdC forms a discontinuous, dynamic ring-like structure in the peripheral membrane of the mother cell, near the mother cell proximal pole of the forespore. A synthetic sporulation screen identified genetic relationships between ssdC and genes involved in the assembly of the spore coat. Phenotypic characterization of these mutants revealed that spore shape, and SsdC localization, depend on the coat basement layer proteins SpoVM and SpoIVA, the encasement protein SpoVID and the inner coat protein SafA. Importantly, we found that the ΔssdC mutant produces spores with an abnormal-looking cortex, and abolishing cortex synthesis in the mutant largely suppresses its shape defects. Thus, SsdC appears to play a role in the proper assembly of the spore cortex, through connections to the spore coat. Collectively, our data suggest functional diversification of the MucB / RseB protein domain between diderm and monoderm bacteria and identify SsdC as an important factor in spore shape development., Author summary Cell shape is an important cellular attribute linked to cellular function and environmental adaptation. Bacterial endospores are one of the toughest cell types on Earth, with a defined shape and complex, highly-resistant, multi-layered cell envelope. Although decades of research have focused on defining the composition and assembly of the multi-layered spore envelope, little is known about how these layers contribute to spore shape. Here, we identify SsdC, a poorly-characterized new member of the MucB / RseB family of proteins that bind lipopolysaccharide in diderm bacteria. We show that SsdC is an important factor in spore shape development in the monoderm, model organism Bacillus subtilis. Our data suggest that SsdC influences the assembly of the spore cortex, through connections to the spore coat, by forming an intriguing, dynamic ring-like structure adjacent to the developing spore. Furthermore, our identification of SsdC suggests evolutionary diversification of the MucB /RseB protein domain between diderm and monoderm bacteria.

Published
2020

21. Human Plasminogen Exacerbates Clostridioides difficile Enteric Disease and Alters the Spore Surface

Author

Awad, Milena, Hutton, Melanie, Quek, Adam, Klare, William, Mileto, Steven, Mackin, Kate, Ly, Diane T, Oorschot, Viola, Bosnjak, Marijana, Jenkin, Grant, Conroy, Paul, West, Nick, Fulcher, Alex, Costin, Adam, Day, Christopher, Jennings, Michael, Dr Robert Medcalf, Robert, Sanderson-Smith, Martina L, Cordwell, Stuart, Law, Ruby, Whisstock, James, Lyras, Dena, Awad, Milena, Hutton, Melanie, Quek, Adam, Klare, William, Mileto, Steven, Mackin, Kate, Ly, Diane T, Oorschot, Viola, Bosnjak, Marijana, Jenkin, Grant, Conroy, Paul, West, Nick, Fulcher, Alex, Costin, Adam, Day, Christopher, Jennings, Michael, Dr Robert Medcalf, Robert, Sanderson-Smith, Martina L, Cordwell, Stuart, Law, Ruby, Whisstock, James, and Lyras, Dena

Abstract

© 2020 Background & Aims: The protease plasmin is an important wound healing factor, but it is not clear how it affects gastrointestinal infection–mediated damage, such as that resulting from Clostridioides difficile. We investigated the role of plasmin in C difficile–associated disease. This bacterium produces a spore form that is required for infection, so we also investigated the effects of plasmin on spores. Methods: C57BL/6J mice expressing the precursor to plasmin, the zymogen human plasminogen (hPLG), or infused with hPLG were infected with C difficile, and disease progression was monitored. Gut tissues were collected, and cytokine production and tissue damage were analyzed by using proteomic and cytokine arrays. Antibodies that inhibit either hPLG activation or plasmin activity were developed and structurally characterized, and their effects were tested in mice. Spores were isolated from infected patients or mice and visualized using super-resolution microscopy; the functional consequences of hPLG binding to spores were determined. Results: hPLG localized to the toxin-damaged gut, resulting in immune dysregulation with an increased abundance of cytokines (such as interleukin [IL] 1A, IL1B, IL3, IL10, IL12B, MCP1, MP1A, MP1B, GCSF, GMCSF, KC, TIMP-1), tissue degradation, and reduced survival. Administration of antibodies that inhibit plasminogen activation reduced disease severity in mice. C difficile spores bound specifically to hPLG and active plasmin degraded their surface, facilitating rapid germination. Conclusions: We found that hPLG is recruited to the damaged gut, exacerbating C difficile disease in mice. hPLG binds to C difficile spores, and, upon activation to plasmin, remodels the spore surface, facilitating rapid spore germination. Inhibitors of plasminogen activation might be developed for treatment of C difficile or other infection-mediated gastrointestinal diseases.

Published
2020

22. Chromosome Segregation and Peptidoglycan Remodeling Are Coordinated at a Highly Stabilized Septal Pore to Maintain Bacterial Spore Development

Author

Mohamed, Ahmed M.T., primary, Chan, Helena, additional, Luhur, Johana, additional, Bauda, Elda, additional, Gallet, Benoit, additional, Morlot, Cécile, additional, Cole, Louise, additional, Awad, Milena, additional, Crawford, Simon, additional, Lyras, Dena, additional, Rudner, David Z., additional, and Rodrigues, Christopher D.A., additional

Published
2021
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24. Perfringolysin O expression in Clostridium perfringens is independent of the upstream pfoR gene

Author

Awad, Milena M. and Rood, Julian I.

Subjects
Pathogenic microorganisms -- Genetic aspects, Gene expression -- Analysis, Genetic transcription -- Regulation, Biological sciences
Abstract

Results point out that the Clostridium perfringens pfoR gene encoded transcriptional activator is not likely to play a role in the activation of the pfoA gene expression. This is corroborated by a mutation in pfoR not altering perfringolysin O production in the bacterrium.

Published
2002

26. Human Plasminogen Exacerbates Clostridioides difficile Enteric Disease and Alters the Spore Surface

Author

Awad, Milena M., primary, Hutton, Melanie L., additional, Quek, Adam J., additional, Klare, William P., additional, Mileto, Steven J., additional, Mackin, Kate, additional, Ly, Diane, additional, Oorschot, Viola, additional, Bosnjak, Marijana, additional, Jenkin, Grant, additional, Conroy, Paul J., additional, West, Nick, additional, Fulcher, Alex, additional, Costin, Adam, additional, Day, Christopher J., additional, Jennings, Michael P., additional, Medcalf, Robert L., additional, Sanderson-Smith, Martina, additional, Cordwell, Stuart J., additional, Law, Ruby H.P., additional, Whisstock, James C., additional, and Lyras, Dena, additional

Published
2020
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27. Tranexamic Acid Influences the Immune Response, but not Bacterial Clearance in a Model of Post-Traumatic Brain Injury Pneumonia

Author

Draxler, Dominik F., primary, Awad, Milena M., additional, Hanafi, Gryselda, additional, Daglas, Maria, additional, Ho, Heidi, additional, Keragala, Charithani, additional, Galle, Adam, additional, Roquilly, Antoine, additional, Lyras, Dena, additional, Sashindranath, Maithili, additional, and Medcalf, Robert L., additional

Published
2019
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28. Additional file 5: of Paeniclostridium sordellii and Clostridioides difficile encode similar and clinically relevant tetracycline resistance loci in diverse genomic locations

Author

Vidor, Callum, Bulach, Dieter, Awad, Milena, and Lyras, Dena

Abstract

Figure S3. Nucleotide alignment of the regions flanking regA in P. sordellii Tet P+ isolates. Sequences for the regA+ isolates SSCC32125, R32462, JGS6961 and SSCC18392 are compared to the regAâ isolate W2945. The direct repeats flanking regA are shown in bold red font and indicated by a black arrow beneath the sequence. The coding region of regA is colored purple. A gap in the alignment representing the majority of regA is indicated by three large dots. The start and stop codons of regA are in bold and annotated on the sequence. The regAâ isolates W3026, AM370 and W2922 have not been included in this alignment; however, a single site identical to the repeat is present for this isolate, equidistant to Tet P, as for W2945. (PDF 238 kb)

Published
2019
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32. Paeniclostridium (Clostridium) sordellii–associated enterocolitis in 7 horses.

Author

Uzal, Francisco A., Navarro, Mauricio A., Hostetter, Jesse M., Nyaoke, Akinyi C., Fresneda, Karina, Diab, Santiago S., Moore, Janet, Lyras, Dena, and Awad, Milena

Subjects
ENTEROCOLITIS, HORSE diseases, HORSES, ANIMAL diseases, CLOSTRIDIUM, INTESTINAL infections, CLOSTRIDIA
Abstract

Enteric disease in horses may be caused by a variety of microorganisms, including several clostridial species. Paeniclostridium sordellii (previously Clostridium sordellii) has been frequently associated with gas gangrene in humans and several animal species, including horses. However, its role in enteric diseases of animals has not been fully determined. We describe herein 7 cases of enteric disease in horses associated with P. sordellii infection. Grossly, the small and/or large intestines were necrotic, hemorrhagic, and edematous. Microscopically, there was severe mucosal necrosis and hemorrhage of the small and/or large intestine of all horses. P. sordellii was isolated and/or demonstrated by immunohistochemistry and/or PCR in the intestine of all horses. All other known causes of enteric disease in horses were ruled out in these 7 cases. P. sordellii should be considered among the differential diagnoses in cases of enteric disease in horses. [ABSTRACT FROM AUTHOR]

Published
2020
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36. The NEAT Domain-Containing Proteins of Clostridium perfringens Bind Heme

Author

Choo, Jocelyn M., primary, Cheung, Jackie K., additional, Wisniewski, Jessica A., additional, Steer, David L., additional, Bulach, Dieter M., additional, Hiscox, Thomas J., additional, Chakravorty, Anjana, additional, Smith, A. Ian, additional, Gell, David A., additional, Rood, Julian I., additional, and Awad, Milena M., additional

Published
2016
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38. Skewed genomic variability in strains of the toxigenic bacterial pathogen, Clostridium perfringens

Author

Myers, Garry S.A., DeBoy, Robert T., Qinghu Ren, Varga, John, Awad, Milena M., Brinkac, Lauren M., Daugherty, Sean C., Rasko, David A., Haft, Daniel H., Dodson, Robert J., Madupu, Ramana, Nelson, William C., Rosovitz, M.J., Sullivan, Steven A., Cheung, Jackie K., Khouri, Hoda, Dimitrov, George I., Watkins, Kisha L., Mulligan, Stephanie, Benton, Jonathan, Radune, Diana, Fisher, Derek J., Atkins, Helens S., Ravel, Jacques, Hiscox, Tom, Billington, Stephen J., Songer, J. Glenn, McClane, Bruce A., Titball, Richard W., Seshadri, Rekha, Rood, Julian I., Melville, Stephen B., and Paulsen, Ian T.

Subjects
Clostridium -- Genetic aspects, Genetic variation -- Research, Toxicogenomics -- Research, Health
Abstract

The complete genome sequences of Clostridium perfringens strain ATCC 13124, a gas gangrene isolate and the species type strain, and the enterotoxin-producing food poisoning strain SM101, were determine and compared with the published C. perfringens strain 13 genome. Comparison of the three genomes revealed considerable genomic diversity with >300 unique 'genomic islands' identified, with the majority of these islands unusually clustered on one replichore.

Published
2006

48. The NanI and NanJ Sialidases of Clostridium perfringens Are Not Essential for Virulence

Author

Chiarezza, Martina, primary, Lyras, Dena, additional, Pidot, Sacha J., additional, Flores-Díaz, Marietta, additional, Awad, Milena M., additional, Kennedy, Catherine L., additional, Cordner, Leanne M., additional, Phumoonna, Tongted, additional, Poon, Rachael, additional, Hughes, Meredith L., additional, Emmins, John J., additional, Alape-Girón, Alberto, additional, and Rood, Julian I., additional

Published
2009
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