Author: "Catherine Booth" - Searchworks@Jio Institute Digital Library Search Results

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136 results on '"Catherine Booth"'

1. Comparison of Antibacterial Activity of Phytochemicals against Common Foodborne Pathogens and Potential for Selection of Resistance

Author: Ryan Sweet, Catherine Booth, Kathryn Gotts, Stephen F. Grove, Paul A. Kroon, and Mark Webber
Subjects: antimicrobial resistance, AMR, phenolics, flavonoids, natural products, Biology (General), QH301-705.5
Abstract: Antimicrobial resistance is now commonly observed in bacterial isolates from multiple settings, compromising the efficacy of current antimicrobial agents. Therefore, there is an urgent requirement for efficacious novel antimicrobials to be used as therapeutics, prophylactically or as preservatives. One promising source of novel antimicrobial chemicals is phytochemicals, which are secondary metabolites produced by plants for numerous purposes, including antimicrobial defence. In this report, we compare the bioactivity of a range of phytochemical compounds, testing their ability to directly inhibit growth or to potentiate other antimicrobials against Salmonella enterica Typhimurium, Pseudomonas aeruginosa, Listeria monocytogenes, and Staphylococcus aureus. We found that nine compounds displayed consistent bioactivity either as direct antimicrobials or as potentiators. Thymol at 0.5 mg/mL showed the greatest antimicrobial effect and significantly reduced the growth of all species, reducing viable cell populations by 66.8%, 43.2%, 29.5%, and 70.2% against S. enterica Typhimurium, S. aureus, P. aeruginosa, and L. monocytogenes, respectively. Selection of mutants with decreased susceptibility to thymol was possible for three of the pathogens, at a calculated rate of 3.77 × 10−8, and characterisation of S. enterica Typhimurium mutants showed a low-level MDR phenotype due to over-expression of the major efflux system AcrAB-TolC. These data show that phytochemicals can have strong antimicrobial activity, but emergence of resistance should be evaluated in any further development.
Published: 2023
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2. Extracellular vesicles produced by the human commensal gut bacterium Bacteroides thetaiotaomicron affect host immune pathways in a cell‐type specific manner that are altered in inflammatory bowel disease

Author: Lejla Gul, Dezso Modos, Sonia Fonseca, Matthew Madgwick, John P. Thomas, Padhmanand Sudhakar, Catherine Booth, Régis Stentz, Simon R. Carding, and Tamas Korcsmaros
Subjects: extracellular vesicles, host‐microbe interactions, single‐cell data analysis, toll‐like receptor pathway, ulcerative colitis, Cytology, QH573-671
Abstract: Abstract The gastrointestinal (GI) tract harbours a complex microbial community, which contributes to its homeostasis. A disrupted microbiome can cause GI‐related diseases, including inflammatory bowel disease (IBD), therefore identifying host‐microbe interactions is crucial for better understanding gut health. Bacterial extracellular vesicles (BEVs), released into the gut lumen, can cross the mucus layer and access underlying immune cells. To study BEV‐host interactions, we examined the influence of BEVs generated by the gut commensal bacterium, Bacteroides thetaiotaomicron, on host immune cells. Single‐cell RNA sequencing data and host‐microbe protein‐protein interaction networks were used to predict the effect of BEVs on dendritic cells, macrophages and monocytes focusing on the Toll‐like receptor (TLR) pathway. We identified biological processes affected in each immune cell type and cell‐type specific processes including myeloid cell differentiation. TLR pathway analysis highlighted that BEV targets differ among cells and between the same cells in healthy versus disease (ulcerative colitis) conditions. The in silico findings were validated in BEV‐monocyte co‐cultures demonstrating the requirement for TLR4 and Toll‐interleukin‐1 receptor domain‐containing adaptor protein (TIRAP) in BEV‐elicited NF‐kB activation. This study demonstrates that both cell‐type and health status influence BEV‐host communication. The results and the pipeline could facilitate BEV‐based therapies for the treatment of IBD.
Published: 2022
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3. Multiparametric MRI of early tumor response to immune checkpoint blockade in metastatic melanoma

Author: Frank Riemer, Pippa G Corrie, Catherine Booth, Ferdia A Gallagher, Kevin M Brindle, Martin J Graves, Arthur Lewis, Doreen Lau, Mary A McLean, Andrew N Priest, Andrew B Gill, Francis Scott, Ilse Patterson, Bruno Carmo, Joshua D Kaggie, Amy Frary, Doreen Milne, Michal Sulikowski, Lee Brown, Jean-Martin Lapointe, and Luigi Aloj
Subjects: Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Published: 2021
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4. Genome Characterization of a Novel Wastewater Bacteroides fragilis Bacteriophage (vB_BfrS_23) and its Host GB124

Author: Mohammad A. Tariq, Fiona Newberry, Rik Haagmans, Catherine Booth, Tom Wileman, Lesley Hoyles, Martha R. J. Clokie, James Ebdon, and Simon R. Carding
Subjects: bacteriophage, Bacteroides fragilis, genomes, waste water, human, Microbiology, QR1-502
Abstract: Bacteroides spp. are part of the human intestinal microbiota but can under some circumstances become clinical pathogens. Phages are a potentially valuable therapeutic treatment option for many pathogens, but phage therapy for pathogenic Bacteroides spp. including Bacteroides fragilis is currently limited to three genome-sequenced phages. Here we describe the isolation from sewage wastewater and genome of a lytic phage, vB_BfrS_23, that infects and kills B. fragilis strain GB124. Transmission electron microscopy identified this phage as a member of the Siphoviridae family. The phage is stable when held at temperatures of 4 and 60°C for 1 h. It has a very narrow host range, only infecting one host from a panel of B. fragilis strains (n = 8). Whole-genome sequence analyses of vB_BfrS_23 determined it is double-stranded DNA phage and is circularly permuted, with a genome of 48,011 bp. The genome encodes 73 putative open reading frames. We also sequenced the host bacterium, B. fragilis GB124 (5.1 Mb), which has two plasmids of 43,923 and 4,138 bp. Although this phage is host specific, its isolation together with the detailed characterization of the host B. fragilis GB124 featured in this study represent a useful starting point from which to facilitate the future development of highly specific therapeutic agents. Furthermore, the phage could be a novel tool in determining water (and water reuse) treatment efficacy, and for identifying human fecal transmission pathways within contaminated environmental waters and foodstuffs.
Published: 2020
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5. The Uptake, Trafficking, and Biodistribution of Bacteroides thetaiotaomicron Generated Outer Membrane Vesicles

Author: Emily J. Jones, Catherine Booth, Sonia Fonseca, Aimee Parker, Kathryn Cross, Ariadna Miquel-Clopés, Isabelle Hautefort, Ulrike Mayer, Tom Wileman, Régis Stentz, and Simon R. Carding
Subjects: Bacteroides thetaiotaomicron, outer membrane vesicles, microvesicles, bacterial extracellular vesicles, gut microbiota, GI-tract, Microbiology, QR1-502
Abstract: Gram-negative bacteria ubiquitously produce and release nano-size, non-replicative outer membrane vesicles (OMVs). In the gastrointestinal (GI-) tract, OMVs generated by members of the intestinal microbiota are believed to contribute to maintaining the intestinal microbial ecosystem and mediating bacteria–host interactions, including the delivery of bacterial effector molecules to host cells to modulate their physiology. Bacterial OMVs have also been found in the bloodstream although their origin and fate are unclear. Here we have investigated the interactions between OMVs produced by the major human gut commensal bacterium, Bacteroides thetaiotaomicron (Bt), with cells of the GI-tract. Using a combination of in vitro culture systems including intestinal epithelial organoids and in vivo imaging we show that intestinal epithelial cells principally acquire Bt OMVs via dynamin-dependent endocytosis followed by intracellular trafficking to LAMP-1 expressing endo-lysosomal vesicles and co-localization with the perinuclear membrane. We observed that Bt OMVs can also transmigrate through epithelial cells via a paracellular route with in vivo imaging demonstrating that within hours of oral administration Bt OMVs can be detected in systemic tissues and in particular, the liver. Our findings raise the intriguing possibility that OMVs may act as a long-distance microbiota–host communication system.
Published: 2020
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6. Comparison of PCR versus PCR-Free DNA Library Preparation for Characterising the Human Faecal Virome

Author: Shen-Yuan Hsieh, Mohammad A. Tariq, Andrea Telatin, Rebecca Ansorge, Evelien M. Adriaenssens, George M. Savva, Catherine Booth, Tom Wileman, Lesley Hoyles, and Simon R. Carding
Subjects: virome, PCR bias, bacteriophage, Microbiology, QR1-502
Abstract: The human intestinal microbiota is abundant in viruses, comprising mainly bacteriophages, occasionally outnumbering bacteria 10:1 and is termed the virome. Due to their high genetic diversity and the lack of suitable tools and reference databases, the virome remains poorly characterised and is often referred to as “viral dark matter”. However, the choice of sequencing platforms, read lengths and library preparation make study design challenging with respect to the virome. Here we have compared the use of PCR and PCR-free methods for sequence-library construction on the Illumina sequencing platform for characterising the human faecal virome. Viral DNA was extracted from faecal samples of three healthy donors and sequenced. Our analysis shows that most variation was reflecting the individually specific faecal virome. However, we observed differences between PCR and PCR-free library preparation that affected the recovery of low-abundance viral genomes. Using three faecal samples in this study, the PCR library preparation samples led to a loss of lower-abundance vOTUs evident in their PCR-free pairs (vOTUs 128, 6202 and 8364) and decreased the alpha-diversity indices (Chao1 p-value = 0.045 and Simpson p-value = 0.044). Thus, differences between PCR and PCR-free methods are important to consider when investigating “rare” members of the gut virome, with these biases likely negligible when investigating moderately and highly abundant viruses.
Published: 2021
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7. Guideline on the investigation and management of acute transfusion reactions

Author: Richard Soutar, Wendy McSporran, Tracey Tomlinson, Catherine Booth, and Sharran Grey
Subjects: Hematology
Published: 2023
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8. The Proteome of Extracellular Vesicles Produced by the Human Gut Bacteria Bacteroides thetaiotaomicron In Vivo Is Influenced by Environmental and Host-Derived Factors

Author: Régis Stentz, Emily Jones, Rokas Juodeikis, Udo Wegmann, Maria Guirro, Andrew J. Goldson, Arlaine Brion, Catherine Booth, Padhmanand Sudhakar, Ian R. Brown, Tamás Korcsmáros, and Simon R. Carding
Subjects: Science & Technology, Ecology, PROTEINS, OUTER-MEMBRANE VESICLES, proteome, COMMUNICATION, Microbiology, Applied Microbiology and Biotechnology, ASPARAGINASE, Bacteroides thetaiotaomicron, Biotechnology & Applied Microbiology, IV, bacterial extracellular vesicles, microbiota, POLYSACCHARIDE UTILIZATION, Life Sciences & Biomedicine, intestine, Food Science, Biotechnology
Abstract: Bacterial extracellular vesicles (BEVs) released from both Gram-negative and Gram-positive bacteria provide an effective means of communication and trafficking of cell signaling molecules. In the gastrointestinal tract (GIT) BEVs produced by members of the intestinal microbiota can impact host health by mediating microbe-host cell interactions. A major unresolved question, however, is what factors influence the composition of BEV proteins and whether the host influences protein packaging into BEVs and secretion into the GIT. To address this, we have analyzed the proteome of BEVs produced by the major human gut symbiont Bacteroides thetaiotaomicron both in vitro and in vivo in the murine GIT in order to identify proteins specifically enriched in BEVs produced in vivo. We identified 113 proteins enriched in BEVs produced in vivo, the majority (62/113) of which accumulated in BEVs in the absence of any changes in their expression by the parental cells. Among these selectively enriched proteins, we identified dipeptidyl peptidases and an asparaginase and confirmed their increased activity in BEVs produced in vivo. We also showed that intact BEVs are capable of degrading bile acids via a bile salt hydrolase. Collectively these findings provide additional evidence for the dynamic interplay of host-microbe interactions in the GIT and the existence of an active mechanism to drive and enrich a selected group of proteins for secretion into BEVs in the GIT. IMPORTANCE The gastrointestinal tract (GIT) harbors a complex community of microbes termed the microbiota that plays a role in maintaining the host's health and wellbeing. How this comes about and the nature of microbe-host cell interactions in the GIT is still unclear. Recently, nanosized vesicles naturally produced by bacterial constituents of the microbiota have been shown to influence responses of different host cells although the molecular basis and identity of vesicle-born bacterial proteins that mediate these interactions is unclear. We show here that bacterial extracellular vesicles (BEVs) produced by the human symbiont Bacteroides thetaiotaomicron in the GIT are enriched in a set of proteins and enzymes, including dipeptidyl peptidases, an asparaginase and a bile salt hydrolase that can influence host cell biosynthetic pathways. Our results provide new insights into the molecular basis of microbiota-host interactions that are central to maintaining GIT homeostasis and health. ispartof: APPLIED AND ENVIRONMENTAL MICROBIOLOGY vol:88 issue:16 ispartof: location:United States status: published
Published: 2022

9. The Proteome of Extracellular Vesicles Produced by the Human Gut Bacteria Bacteroides thetaiotaomicron

Author: Régis, Stentz, Emily, Jones, Rokas, Juodeikis, Udo, Wegmann, Maria, Guirro, Andrew J, Goldson, Arlaine, Brion, Catherine, Booth, Padhmanand, Sudhakar, Ian R, Brown, Tamás, Korcsmáros, and Simon R, Carding
Subjects: Bacteroides thetaiotaomicron, Extracellular Vesicles, Mice, Bacteria, Proteome, Animals, Asparaginase, Humans, Dipeptidyl-Peptidases and Tripeptidyl-Peptidases, Gastrointestinal Microbiome
Abstract: Bacterial extracellular vesicles (BEVs) released from both Gram-negative and Gram-positive bacteria provide an effective means of communication and trafficking of cell signaling molecules. In the gastrointestinal tract (GIT) BEVs produced by members of the intestinal microbiota can impact host health by mediating microbe-host cell interactions. A major unresolved question, however, is what factors influence the composition of BEV proteins and whether the host influences protein packaging into BEVs and secretion into the GIT. To address this, we have analyzed the proteome of BEVs produced by the major human gut symbiont Bacteroides thetaiotaomicron both
Published: 2022

10. Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial

Author: Obbina Abani, Ali Abbas, Fatima Abbas, Mustafa Abbas, Sadia Abbasi, Hakam Abbass, Alfie Abbott, Nabeel Abdallah, Ashraf Abdelaziz, Mohamed Abdelfattah, Bushra Abdelqader, David Abdo, Basir Abdul, Althaf Abdul Rasheed, Ajibode Abdulakeem, Rezan Abdul-Kadir, Amina Abdulle, Abdulfatahi Abdulmumeen, Rasheed Abdul-Raheem, Niyaz Abdulshukkoor, Kula Abdusamad, Yazeed Abed El Khaleq, Mai Abedalla, Abeer Ul Amna Abeer Ul Amna, Katrina Abernethy, Adebanke Aboaba, Hani Abo-Leyah, Ahmed Abou-Haggar, Mahmoud Abouibrahim, Miriam Abraham, Tizzy Abraham, Abraheem Abraheem, Judith Abrams, Hyacinth-John Abu, Ahmed Abu-Arafeh, Syed M Abubacker, Akata Abung, Yaa Aceampong, Amaka Achara, Devikumar Acharya, Sarah Acheampong, Janet Acheson, Andres Acosta, Catherine Acton, Jacqueline Adabie-Ankrah, Sara Adair, Fiona Adam, Matthew Adam, Huzaifa Adamali, Carol Adams, Charlotte Adams, Kate Adams, Lisa Adams, Richard Adams, Tim Adams, Kirsty Adcock, Jemaimah Addai, Ade Adebiyi, Ken Adegoke, Vicki Adell, Debo Ademokun, Sherna Adenwalla, Oluwasegun A Adesemoye, Emmanuel O Adewunmi, Joyce Adeyemi, Rina Adhikary, Gabrielle Adkins, Adnan Adnan, John Aeron-Thomas, Debbie Affleck, Dominic Affron, Carmel Afnan, Muhammad Afridi, Zainab A Aftab, Meenakshi Agarwal, Rachel Agbeko, Chris Agbo, Penny Agent, Sunil Aggarwal, Arameh Aghababaie, Shafana Ahamed Sadiq, Mohamed H Ahammed Nazeer, Humayun Ahmad, Mohammad Ahmad, Syed Ahmad, Asim Ahmed, Bilal Ahmed, Forizuddin Ahmed, Hamze Ahmed, Iram Ahmed, Irshad Ahmed, Khaled Ahmed, Liban Ahmed, Mahin Ahmed, Maria C Ahmed, Muhammad S Ahmed, Naseer Ahmed, Nausheen Ahmed, Osama Ahmed, Rajia A Ahmed, Rizwan Ahmed, Saif Ahmed, Sammiya Ahmed, Sara Ahmed, Sophia Ahmed, Syed Ahmed, Syed Haris Ahmed, Roa Ahmed Ali, Sana Ahmed, Sana Ahmer, Dhiraj Ail, Mark Ainsworth, Giulia Airoldi, Myriam Aissa, Lindianne Aitken, Bini Ajay, Francis Ajeneye, Abdulakeem Ajibode, Ayesha Ajmi, Tahir Akbar, Naim Akhtar, Nasim Akhtar, Nauman Akhtar, Suha Akili, Oludoyinsola Akindolie, Yinka Akinfenwa, Olugbenga Akinkugbe, Ibrahim Akinpelu, Olajide Akinwumiju, Olugbenro Aktinade, Ahmad Al Aaraj, Asma Al Balushi, Majd Al Dakhola, Aladdin Al Swaifi, Eslam Al-Abadi, Narendra Aladangady, Ayaz Alam, Sajid Alam, Abbas Al-Asadi, Karina Al-Asadi, Kyriaki Alatzoglou, Manaf Al-Bayati, Paul Albert, Lorraine Albon, Gemma Alcorn, Stephen Alcorn, Aggie Aldana, David Alderdice, Rayan Aldouri, Jonathan Aldridge, Nicolas Aldridge, Ana Alegria, Alison Alexander, John Alexander, Peter D G Alexander, Charlotte Alford, Julyan Al-Fori, Laith Alghazawi, Bahij Al-Hakim, Shams Al-Hity, Ali Ali, Asad Ali, Fawzia R Ali, Hoodo Ali, Jawad Ali, Mariam Ali, Mohammad Ali, Nayab Ali, Oudai Ali, Sabira Ali, Sakina Ali, Syed Ali, Abid Alina, Fine Aliyuda, Katrin Alizadeh, Maithem Al-Jibury, Saba Al-Juboori, Majid Al-Khalil, Moutaz Alkhusheh, Allameddine Allameddine, Fiona Allan, Rachel Allan, Alison Allanson, Robert Allcock, Beverley Allen, Eireann Allen, Jess Allen, Kerry Allen, Laura Allen, Louise Allen, Poppy Allen, Rebecca Allen, Sam Allen, Sharon Allen, Simon Allen, Kathryn Allison, Bethan Allman, Lynne Allsop, Hassan Al-Moasseb, Magda Al-Obaidi, Lina Alomari, Akram Al-Rabahi, Bahar Al-Ramadhani, Zayneb Al-Saadi, Inji Alshaer, Rustam Al-Shahi Salman, Warkaq Al-Shamkhani, Bashar Al-Sheklly, Sara Altaf, Mary Alvarez, Balaal Alyas, Maysaa Alzetani, Susan Amamou, Noor Amar, Sakkarai Ambalavanan, Sarah-Jayne Ambler, Robert Ambrogetti, Chris Ambrose, Amir Ameen, Kenneth Amenyah, Maria R Amezaga, Allison Amin, Amina Amin, Kanish Amin, Syed Amin, Tara Amin, Amjad Amjad, Neelma Amjad, Mariam Ammoun, Victoria Amosun, Khaled Amsha, Pugh Amy, Atul Anand, Rekha Anand, Samantha Anandappa, Julie Anderson, Kevin Anderson, Laura Anderson, Michelle Anderson, Nicola Anderson, Rachel Anderson, Rory Anderson, Wendy Anderson, Prematie Andreou, Angela Andrews, Antonette Andrews, Jill Andrews, Susan Andrews, Gregory Andrikopoulos, Kanayochukwu Aneke, Andrew Ang, Wan Wei Ang, Tammy Angel, Aramburo Angela, Paola Angelini, Lazarus Anguvaa, Oleg Anichtchik, Millicent Anim-Somuah, Krishnan Aniruddhan, Jessica Annett, Patrick J Anstey, Rebekah Anstey, Alpha Anthony, Aaron Anthony-Pillai, Philip Antill, Zhelyazkova Antonina, Varghese Anu, Muhammad Anwar, George Apostolides, Aristeidis Apostolopoulos, Sarah Appleby, Diane Appleyard, Maia Far Aquino, Bianca Araba, Samuel Aransiola, Mariana Araujo, Emily Arbon, Ann Archer, Denise Archer, Simon Archer, Christian Ardley, Ana-Maria Arias, Ryoki Arimoto, Charlotte Arkley, Charlotte Armah, Ilianna Armata, Adam Armitage, Ceri Armstrong, Maureen Armstrong, Sonia Armstrong, Sylvia Armstrong-Fisher, Philippa Armtrong, Heike Arndt, Clare Arnison-Newgass, David Arnold, Rachael Arnold, Sarah Arnott, Dhawal Arora, Kavan Arora, Pardeep Arora, Rishi Arora, Andrea Arroyo, Arslam Arter, Ayush Arya, Rita Arya, Denisa Asandei, Adeeba Asghar, Catherine Ashbrook-Raby, Glen Ashby, Helen Ashby, Jan Ashcroft, John Ashcroft, Samuel Ashcroft, Deborah Asher, Ayesha Ashfaq, Ben Ashford, Suhail Ashgar, Abdul Ashish, Sally Ashman-Flavell, Sundar Ashok, Abd-El-Aziz Ashour, Muhammad Z Ashraf, Saima Ashraf, Mohammad B Ashraq, Deborah Ashton, Susan Ashton, Andrew Ashworth, Rebecca Ashworth, Arshia Aslam, Harshini Asogan, Abigail Asquith, Atif Asrar, Omar Assaf, Raine Astin-Chamberlain, Richard Athay-Hunt, Deborah Athorne, Billie Atkins, Christopher Atkins, Stacey Atkins, John Atkinson, Vicki Atkinson, Brygitta Atraskiewicz, Claire Atterbury, Abdul Ahmad Attia, Rita Atugonza, Paula Aubrey, Avinash Aujayeb, Aye Chan Thar Aung, Hnin Aung, Kyaw Thu Aung, Ni Aung, Yin Aung, Zaw Myo Aung, Emily Austin, Karen Austin, Abdusshakur Auwal, Miriam Avery, Joanne Avis, Georgina Aviss, Cristina Avram, Paula Avram, Gabriel Awadzi, Atia Awan, Aszad Aya, Eman Ayaz, Amanda Ayers, Jawwad Azam, Mohammed Azharuddin, Ghazala Aziz, N Aziz, Ali Azkoul, Ashaari Azman Shah, Giada Azzopardi, Hocine Azzoug, Fiyinfoluwa Babatunde, Melvin Babi, Babiker Babiker, Gayna Babington, Matthew Babirecki, Marta Babores, Adetona O Babs-Osibodu, Sammy Bacciarelli, Roudi Bachar, Gina Bacon, Jenny Bacon, Bibi Badal, Gurpreet R Badhan, Shreya Badhrinarayanan, Joseph P Bae, Sibel Bafekr Mishamandani, Alice Baggaley, Amy Baggott, Graham Bagley, Dinesh Bagmane, Lynsey Bagshaw, Kasra Bahadori, James Bailey, Katie Bailey, Lindsey Bailey, Liz Bailey, Morgan Bailey, Pippa Bailey, Sarah Bailey, Stephen Bailey, Hamish Baillie, J Kenneth Baillie, Jennifer Bain, Sanchia Baines, Vikram Bains, Aimi Baird, David Baird, Susan Baird, Tracy Baird, Yolanda Baird, Aiysha Bajandouh, Charles Baker, Emma Baker, Evelyn Baker, Johanne Baker, Josephine Baker, Kenneth Baker, Rebecca Baker, Terri-Anne Baker, Victoria Baker, Hugh Bakere, Nawar Bakerly, Michelle Baker-Moffatt, Nauman Bakhtiar, Panos Bakoulas, Julie Balaam, Niranjan Balachandran, Andrea Balan, Theodosios Balaskas, Madhu Balasubramaniam, Alison Balcombe, Alexander Baldwin, Ashley Baldwin, Caron Baldwin, Danielle Baldwin, Rebekah Baldwin-Jones, James Balfour, Gagan Bali, Sonya Balkee, Ceri Ball, Kasia Ballard, Amy Ballinger, Ismael Balluz, Craig Balmforth, Emese Balogh, Amir Baluwala, Gabby Bambridge, Alasdair Bamford, Amy Bamford, Peter Bamford, Adefunke Bamgboye, Elizabeth Bancroft, Hollie Bancroft, Tanya Bancroft, Joyce Banda, Krishna Bandaru, Srini Bandi, Nageswar Bandla, Somaditya Bandyopadhyam, Amit Banerjee, Millie Banerjee, Ritwik Banerjee, Harrison Banks, Lauren Banks, Luke Banks, Paul Banks, Oliver Bannister, Bharat Bansal, Robert Banthorpe, Laura Banton, Mariamma Baptist, Tanya Baqai, Ananya Mouli Baral, Desislava Baramova, Alex Barber, Russel Barber, Emma Barbon, Miriam Barbosa, Monica Barbosa, Jamie Barbour, Alexander Barclay, Claire Barclay, George Bardsley, Stephanie Bareford, Shahedal Bari, Amy Barker, Debbie Barker, Helen Barker, Joseph Barker, Leon Barker, Oliver Barker, Kerry Barker-Williams, Sinha Barkha, Juliana Barla, Gavin Barlow, Richard Barlow, Valerie Barlow, James Barnacle, Alex Barnard, Debi Barnes, Nicky Barnes, Theresa Barnes, Calum Barnetson, Amy Barnett, Matthew Barnett, Ashton Barnett-Vanes, William Barnsley, Andrew Barr, David Barr, Shaney Barratt, Manuella Barrera, Amy Barrett, Fiona Barrett, Jessica Barrett, Joe Barry, Jazz Bartholomew, Claire Bartlett, Georgina Bartlett, Greg Barton, Jill Barton, Lorna Barton, Rachael Barton, Rosaleen Baruah, Sonia Baryschpolec, Archana Bashyal, Betsy Basker, Ayten Basoglu, Gilda Bass, John Bassett, G Bassett, Chris Bassford, Pavinder Bassi, Betsy Bassis, Bengisu Bassoy, Victoria Bastion, Anupam Basumatary, Tristan Bate, Harry J Bateman, Ian Bateman, Kathryn Bateman, Vhairi Bateman, Eleanor Bates, Hayley Bates, Michelle Bates, Simon Bates, Sally Batham, Ana Batista, Amit Batla, Dushyant Batra, Donna Batty, Harry Batty, Thomas Batty, Peter Baughan, Miranda Baum, Carina Bautista, Fareha Bawa, Fatima S Bawani, Simon Bax, Lydia Baxter, Matt Baxter, Nicola Baxter, Zachary Baxter, Hannah Bayes, Charlotte Baylem, Lee Bayliss, Eileen Bays, Farid Bazari, Rohit Bazaz, Ahmad Bazli, Laura Beacham, Wendy Beadles, Philip Beak, Andy Beale, Kirk Beard, Jack Bearpark, Karen Beaumont, Dawn Beaumont-Jewell, Theresa Beaver, Sarah Beavis, Christy Beazley, Sarah Beck, Virginia Beckett, Rosie Beckitt, Heidi Beddall, Seonaid Beddows, Deborah Beeby, Gail Beech, Michelle Beecroft, Sally Beer, Jane Beety, Gabriela Bega, Alison Begg, Susan Begg, Sara Beghini, Ayesha Begum, Salman Begum, Selina Begum, Teresa Behan, Jasmine Beharry, Roya Behrouzi, Jon Beishon, Claire Beith, James Belcher, Holly Belfield, Katherine Belfield, Ajay Belgaumkar, Dina Bell, Gareth Bell, Gill Bell, Gillian Bell, Joshua Bell, Lauren Bell, Louise Bell, Nicholas Bell, Pippa Bell, Stephanie Bell, Jennifer L Bell, Jennifer Bellamu, Mary Bellamy, Arianna Bellini, Amanda Bellis, Fionn Bellis, Lesley Bendall, Naveena Benesh, Nicola Benetti, Leonie Benham, Guy Benison-Horner, Alexander Bennett, Ann Bennett, Caroline Bennett, Christopher Bennett, Gillian Bennett, Ian Bennett, Kristopher Bennett, Lorraine Bennett, Sara Bennett, Karen Bennion, Vivienne Benson, Jane Benstead, Andrew Bentley, Dionne Bentley, James Bentley, Ian Benton, Eva Beranova, Matthew Beresford, Colin Bergin, Malin Bergstrom, Jolanta Bernatoniene, Thomas Berriman, Zoe Berry, Marnie Berwick, Kimberley Best, Ans-Mari Bester, Yvonne Beuvink, Emily Bevan, Sarah Bevins, Tom Bewick, Andrew Bexley, Sonay Beyatli, Fenella Beynon, Arjun Bhadi, Sanjay Bhagani, Shiv Bhakta, Rekha Bhalla, Khushpreet Bhandal, Kulbinder Bhandal, Ashwin Bhandari, Sangam Bhandari, Aashutosh Bhanot, Ravina Bhanot, Sruti Bhaskara, Prashanth Bhat, Nikhil Bhatia, Rahul Bhatnagar, Karan Bhatt, Janki Bhayani, Deepika Bhojwani, Salimuzzaman Bhuiyan, Anna Bibby, Fatima Bibi, Naheeda Bibi, Salma Bibi, Tihana Bicanic, Sarah Bidgood, Julie Bigg, Sarah Biggs, Alphonsa Biju, Andras Bikov, Sophie Billingham, Jessica Billings, Carron Bilton, Teodirico Binas, Martin Binney, Alice Binns, Muhammad BinRofaie, Oliver Bintcliffe, Catherine Birch, Jenny Birch, Louann Birch, Janet Birchall, Katherine Birchall, Sam Bird, Sumedha Bird, Charndip Biring, Mark Birt, Raquel Bisa, Kilanalei Bishop, Linda Bishop, Lisa Bishop, Karen Bisnauthsing, Nibedan Biswas, Sahar Biuk, Karen Blachford, Ethel Black, Helen Black, Karen Black, Mairead Black, Polly Black, Hayley Blackgrove, Bethan Blackledge, Joanne Blackler, Samantha Blackley, Helen Blackman, Stuart Blackmore, Caroline Blackstock, Loraine Blackwood, Francesca Blakemore, Helen Blamey, Alison Bland, Sujata Blane, Simon Blankley, Mary Blanton, Parry Blaxill, Jenny Blaxland, Katie Blaylock, Jane Blazeby, Carol Blears, Natalie Blencowe, Donna Blofield, Ben Bloom, Jack Bloomfield, Angela Bloss, Hannah Bloxham, Louise Blundell, Andrew Blunsum, Mark Blunt, Ian Blyth, Kevin Blyth, Andrew Blythe, Karen Blythe, Marilyn Boampoaa, Boniface Bobie, Karen Bobruk, Pritesh Bodalia, Neena Bodasing, Tanya Bodenham, Sherin Bodh, Gabriele Boehmer, Marta Boffito, Kristyna Bohmova, Sumit Bokhandi, Maria Bokhar, Saba Bokhari, Sakina Bokhari, Syed Owais Bokhari, Ambrose Boles, Sarah Bollington, Sam Bolton, Charlotte Bomken, Charlotte Bond, Hayley Bond, Helena Bond, Stuart Bond, Thomas Bond, Alice Bone, Georgia Boniface, Wendy Bonnert, Lizzy Bonney, Leigh Boorman, Catherine Booth, Joanne Borbone, Cameron Borhani, Naomi Borman, Rachel Borrell, Mamu Boshir, Fiona Bottrill, Laura Bough, Hayley Boughton, Zoe Boult, Miriam Bourke, Stephen Bourke, Michelle Bourne, Rachel Bousfield, Lucy Boustred, Darren Bowen, Kaye Bowen, Alexandra Bowes, Amy Bowes, Philip Bowker, Louise Bowman, Simon Bowman, Rachel Bowmer, Angie Bowring, Geoff Bowyer, Helen Bowyer, Carmel Boyd, Jenny Boyd, Laura Boyd, Maxine Boyd, Namoi Boyle, Pauline Boyle, Rosalind Boyle, Louise Boyles, Osman Bozdag, Leanna Brace, David Brack, Charlotte Brackstone, Rob Bradburn, Jodie Bradder, Barry Bradley, Clare Jane Bradley, Pamela Bradley, Patrick Bradley, Paul Bradley, Joanne Bradley-Potts, Lynne Bradshaw, Sarah Bradshaw, Zena Bradshaw, Rebecca Brady, Shirin Brady, Denise Braganza, Suhail Brailsford, Jill Braithwaite, Marie Branch, Thomas Brankin-Frisby, Jamie Brannigan, Louise Brassington, Sophie Brattan, Fiona Bray, Nancy Bray, Angela Brazier, Manny Brazil, Lucy Brear, Tracy Brear, Stephen Brearey, Laura Bremner, Morwenna Brend, Giovanna Bretland, Chris Brewer, Hannah Bridge, Gavin Bridgwood, Hayley Briggs, Mark Briggs, Sara Brigham, John Bright, Chris Brightling, Lutece Brimfield, Elaine Brinkworth, Robin Brittain-Long, Vianne Britten, Terri Brittin, Lauren Broad, Sarah Broad, Rosie Broadhurst, Andrew Broadley, Marie Broadway, Christopher Brockelsby, Megan Brocken, Tomos Brockley, Andrew Broderick, Mary Brodsky, Fiona Brogan, David Broggio, Liz Brohan, Felicity Brokke, Jacob Brolly, David Bromley, Hannah Brooke-Ball, Verity Brooker, Ceri Brookes, Matthew Brookes, Alison Brooks, Karen Brooks, Nicole Brooks, Philip Brooks, Rachel Brooks, Sophie Brooks, Natalie Broomhead, Chloe Broughton, Nathaniel Broughton, Matt Brouns, Marie Browett, Alison Brown, Ammani Brown, Carly Brown, Catrin Brown, Chloe Brown, Elizabeth Brown, Ellen Brown, Heather Brown, Janet Brown, Louise Brown, Niall Brown, Pauline Brown, Rachel Brown, Richard Brown, Robert Brown, Steven Brown, Tom Brown, Wayne Brown, Bria Browne, Charlotte Browne, Duncan Browne, Mitchell Browne, Stephen Brownlee, Alba Brraka, David Bruce, Johanna Bruce, Michelle Bruce, Wojciech Brudlo, Nigel Brunskill, Alan Brunton, Margaret Brunton, Mandy Bryan, Meera Bryant, April Buazon, Maya Buch, Julie Buchan, Ruaridh Buchan, Alexis Buchanan, Ruaridh Buchanan, Danielle Buche, Amanda Buck, Matthew Buckland, Laura Buckley, Philip Buckley, Sarah Buckley, Carol Buckman, Kathleen Buckmire, George Bugg, Ramadan Bujazia, Marwan Bukhari, Shanze Bukhari, Richard Bulbulia, Alex Bull, Damian Bull, Rhian Bull, Thomas Bull, Naomi Bulteel, Kasun Bumunarachchi, Roneleeh Bungue-Tuble, Caroline Burchett, Dorota Burda, Christy Burden, Thomas G Burden, Mika Burgess, Paula Burgess, Richard Burgess, Sophia Burgess, Paula Burgett, Adrian Burman, Sara Burnard, Caroline Burnett, John Burnett, Amanda Burns, Amy Burns, Collette Burns, James Burns, Karen Burns, Samuel Burns, Sarah Burns, Daniel Burrage, Sadie Burrage, Kate Burrows, Claire Burston, Ben Burton, Fiona Burton, Matthew Burton, Angela Busby, Deborah Butcher, Aaron Butler, Jessica Butler, Joanne Butler, Joshua Butler, Lesley Butler, Peter Butler, Susan Butler, Al-Tahoor Butt, Mohammad M Butt, Sophia Butt, Caryl Butterworth, Nicola Butterworth-Cowin, Robert Buttery, Tom Buttle, Heather Button, Daniel Buttress, Jane Byrne, Wendy Byrne, Victoria Byrne-Watts, Eleanor Byworth, Amanda Cabandugama, Ruth Cade, Anthony Cadwgan, Donna Cairney, James Calderwood, Darren Caldow, Moira Caldwell, Giorgio Calisti, Debbie Callaghan, Jennifer Callaghan, Claire Callens, Donaldson Callum, Caroline Calver, Melissa Cambell-Kelly, Tracey Camburn, David R Cameron, Eleanor Cameron, Fraser Cameron, Sarah Cameron, Sheena Cameron, Christian Camm, Renee F D Cammack, Alison Campbell, Amy Campbell, Barbara Campbell, Bridget Campbell, Debbie Campbell, Helen Campbell, Hilary Campbell, Jonathan Campbell, Mark Campbell, Robyn Campbell, Wynny Campbell, Quentin Campbell Hewson, Julie Camsooksai, Ana Canabarro, Lisa Canclini, Shaula Mae Candido, Janie Candlish, Cielito Caneja, Johnathon Cann, Ruby Cannan, Emma Cannon, Michael Cannon, Petra Cannon, Vivienne Cannons, Jane Cantliff, Ben Caplin, Santino Capocci, Noemi Caponi, Angelika Capp, Anne Capps-Jenner, Thomas Capstick, Ishmael Carboo, Nuria Cardenas, Mary Cardwell, Rachel Carey, Simon Carley, Tammy Carlin, Andrew Carlton, Samantha Carmichael, Mandy Carnahan, Rebecca Carnegie, Charlotte Caroline, Emily Carpenter, Jodi Carpenter, David Carr, Sharon Carr, Anna Carrasco, Samantha Carrington, Zoe Carrington, Paul Carroll, Caroline Carron, Anne Carstairs, Jonathan Carter, Michael Carter, Moira Carter, Paul Carter, Penny Carter, Steven Carter, Simon Carter-Graham, Douglas Cartwright, Jo-Anne Cartwright, Claire Carty, Sinead Carty, Jaime Carungcong, Carolyn Carveth-Marshall, Susan Casey, Annie Cassells, Barbara Cassimon, Teresa Castiello, Gail Castle, Bridget Castles, Melanie Caswell, Ana Maria Catana, Heidi Cate, Susanne Cathcart, Katrina Cathie, Christine Catley, Laura Catlow, Matthew Caudwell, Jill Caulfield, Anna Cavazza, Luke Cave, Simon Cavinato, Frianne Cawa, Kathryn Cawley, Chloe Caws, Hankins Cendl, Hannah Century, Jeva Cernova, Mansur Cesay, Ed Cetti, Stephanie Chabane, Manish Chablani, Cathleen Chabo, David Chadwick, Julie Chadwick, Robert Chadwick, Ela Chakkarapani, Arup Chakraborty, Mallinath Chakraborty, Mollika Chakravorty, James Chalmers, Richard Chalmers, Georgina Chamberlain, Sarah Chamberlain, Emma Chambers, Jonathan Chambers, Lucy Chambers, Naomi Chambers, Alex Chan, Carmen Chan, Cheuk Chan, Evelyn Chan, Kayen Chan, Kimberley Chan, Ping Chan, Rebekah (Pui-Ching) Chan, Xin Hui Chan, Chris Chandler, Heidi Chandler, Kim J Chandler, Stuart Chandler, Zoe Chandler, Vikki Chandler-Vizard, Sumit Chandra, Navin Chandran, Badrinathan Chandrasekaran, Cherry Chang, Yvonne Chang, Josephine Chaplin, Graeme Chapman, John Chapman, Katie Chapman, Laura Chapman, Lianne Chapman, Polly Chapman, Timothy Chapman, Lucy Chappell, Linda Chapple, Amanda Charalambou, Bethan Charles, Dianne Charlton, Sally Charlton, Kevin Chatar, Calvin Chatha, Ritesh Chaube, Muhammad YN Chaudhary, Iram Chaudhry, Nazia Chaudhuri, Muhammad Chaudhury, Anoop Chauhan, Ruchi S Chauhan, Vipul Chauhan, Nicola Chavasse, Iknam Chaven, Rosanna Chavez, Vipal Chawla, Maria Cheadle, Lindsay Cheater, James Cheaveau, Charlotte Cheeld, Michelle Cheeseman, Fang Chen, Hui Min Chen, Terence Chen, Lok Yin Cheng, Zhihang Cheng, Helen Chenoweth, Chun How Cheong, Shiney Cherian, Suzanne Cherif, Mary Cherrie, Helen Cheshire, Barry Chesterson, Betty Cheung, Chee Kay Cheung, Elaine Cheung, Michelle Cheung, Claire Cheyne, Swati Chhabra, Wei Ling Chia, Eric Chiang, Angela Chiapparino, Rosavic Chicano, Zviedzo A Chikwanha, Sam Chilcott, Phillipa Chimbo, KokWai Chin, Wen Jie Chin, Rumbidzai Chineka, Amol Chingale, Karen Chinn, Vashira Chiroma, Heather Chisem, Claire Chisenga, Ben Chisnall, Carolyn Chiswick, Sunder Chita, Nihil Chitalia, Matthew Chiu, Brenda Chivima, Catherine Chmiel, Soha Choi, Willy Choon Kon Yune, Vandana Choudhary, Sarah Choudhury, Bing-Lun Chow, Fateha Chowdhury, Mahibbur Chowdhury, Shahid Chowdhury, Victoria Christenssen, Peter Christian, Alexander Christides, Fiona Christie, Daniel Christmas, Thereza Christopherson, Mark Christy, Paris Chrysostomou, Yunli Chua, Shabs Chucha, Dip Chudgar, Richard Chudleigh, Srikanth Chukkambotla, Michael E Chukwu, Izu Chukwulobelu, Chi Y Chung, Ben Church, Elaine Church, Sara R Church, David Churchill, Nicole Cianci, Nick Ciccone, Paola Cicconi, Paola Cinardo, Zdenka Cipinova, Bessie Cipriano, Sarah Clamp, Melanie Clapham, Edel Clare, Sarbjit Clare, Andrew Clark, Charlotte Clark, Diane Clark, Felicity Clark, Gabrielle Clark, James Clark, Katherine Clark, Kaylea Clark, Louise Clark, Lucy Clark, Matthew Clark, Patricia Clark, Richard Clark, Thomas Clark, Wendy Clark, Zoe Clark, Andrea Clarke, Heather Clarke, Mark Clarke, Paul Clarke, Robert Clarke, Roseanne Clarke, Samantha Clarke, Sarah Clarke, Sheron Clarke, Tracy Clarke, Andrew Clarkson, Alleyna Claxton, Kate Clay, Elizabeth Clayton, Olivia Clayton, Jill Clayton-Smith, Chris Cleaver, Carlota Clemente de la Torre, Jayne Clements, Suzanne Clements, Francesca Clemons, Lee Clifford, Lynne Clifford, Rachael Clifford, Sarah Clifford, Amelia Clive, Jonathan Clouston, Samantha Clueit, Andrea Clyne, Michelle Coakley, Peter G L Coakley, Tony Coates, Kathryn Cobain, Alexandra Cochrane, Patricia Cochrane, Maeve Cockerell, Helen Cockerill, Shirley Cocks, Rachel Codling, Adam Coe, Samantha Coetzee, David Coey, Danielle Cohen, Jonathan Cohen, Oliver Cohen, Mike Cohn, Louise Coke, Olutoyin Coker, Nicholas Colbeck, Roghan Colbert, Carol Cole, Esther Cole, Jade Cole, Joby Cole, Julie Cole, Richard Cole, Sue Cole, Garry Coleman, Matt Coleman, Holly Coles, Rebecca Coles, Macleod Colin, Alicia Colino-Acevedo, Julie Colley, Dawn Collier, Heather Collier, Paul Collini, Emma Collins, Jaimie Collins, Joanne Collins, Nicola Collins, Sally Collins, Vicky Collins, Andrew Collinson, Bernadette Collinson, Jennifer Collinson, Matthew Collis, Madeleine Colmar, Hayley E Colton, James Colton, Katie Colville, Carolyn Colvin, Ryan Colwell, Edward Combes, David Comer, Alison Comerford, Mike Comery, Dónal Concannon, Robin Condliffe, Lynne Connell, Natalie Connell, Karen Connelly, Gavin Connolly, Mireille Connolly, Emma Connor, Samantha Conran, Antonia Conroy, Veronica Conteh, Rory Convery, Camilla Conway, Francesca Conway, Grainne Conway, Rhiannon Conway, Jo-Anna Conyngham, Colette Cook, Eloise Cook, Gemma Cook, Helen Cook, Julie Cook, Danielle Cooke, Graham Cooke, Katrina Cooke, Soo Cooke, Tim Cooke, Adele Cooper, Alison Cooper, Chris Cooper, David Cooper, Helen Cooper, Jamie Cooper, Joanne Cooper, Joshua Cooper, Laura Cooper, Lauren Cooper, Nick Cooper, Rowena Cooper, Emma Cooperwaite, Thomas Cope, Sinead Corbet, Carolyn Corbett, John Corcoran, Chris Cordell, Jessica Cordle, Alasdair Corfield, John Corless, Alison Corlett, Joe Cornwell, Michael Cornwell, Diana Corogeanu, Mirella Corredera, Ruth Corrigan, Catherine Corry, Rita Corser, Denise Cosgrove, Tracey Cosier, Patricia Costa, Charlie Coston, Susannah Cotgrove, Zoe Coton, Lisa-Jayne Cottam, Rhiannon Cotter, Donna Cotterill, Alice Cotton, Caroline Cotton, Katy Cotton, Andrew Coull, James Coulson, David Counsell, David Counter, Cherry Coupland, Ellie Courtney, Julia Courtney, Rebecca Cousins, Alexander Cowan, Elena Cowan, Ruth Cowburn, Richard Cowell, Louise Cowen, Steve Cowman, Amanda Cowton, Debra Anne Cox, Ellie Cox, Giles Cox, Karina Cox, Miriam Cox, Karen Coy, Beverly Craig, Victoria Craig, Felicity Craighead, Matthew Cramp, Beverley Crane, Jacolene Crause, Adrian Crawford, Angie Crawford, Emma Crawford, Isobel Crawford, Richard Crawforth, Sarah Crawshaw, Ben Creagh-Brown, Andrew Creamer, Ryan Creighton, Joanne Cremona, Saveria Cremona, Janet Cresswell, Mark Cribb, Charles Crichton, Declan Crilly, Lauren Crisp, Nikki Crisp, Dominic Crocombe, Maria Croft, Ian Cron, Derrick Crook, Jennifer Crooks, Helen Croot, Harriet Crosby, Sarah Cross, Tim Cross, Amy Crothers, Stephen Crotty, Susan Crouch, Madeleine Crow, Amanda Crowder, Kate Crowley, Teresa Crowley, Rebecca Croysdill, Callum Cruickshank, Conor Cruickshank, Irena Cruickshank, James Cruise, Helen Crumley, Carina Cruz, Trino Cruz Cervera, Dominic Cryans, Guanguo Cui, Helen Cui, Donna Cullen, Lorraine Cullen, Gillian Cummings-Fosong, Marie Cundall, Victoria Cunliffe, Lorraine Cunningham, Neil Cunningham, Nicola Cunningham, Jason Cupitt, Hollie Curgenven, Debra Curley, Gerens Curnow, David Curran, Simon Curran, Craig Currie, Jacqueline Currie, Scarlett Currie, Abi Curtis, Becca Curtis, Jonathan Curtis, Katrina Curtis, Olivia Curtis, Thomas Curtis, Rebecca Cuthbertson, Sean Cutler, Marta Czekaj, Patrycja Czylok, Joana da Rocha, Andrew Dagens, Helen Daggett, Phil Daggett, Jacqui Daglish, Sandeep Dahiya, Helen Dakers-Black, Anne Dale, Katie Dale, Michaela Dale, Sam Dale, Jolyon Dales, Helen Dalgleish, Nikki Dallas, Helen Dallow, Dermot Dalton, Zoe Daly, Akila Danga, Amelia Daniel, Priya Daniel, Allison Daniels, Adela Dann, Sandra Danso-Bamfo, Nimo Daoud, Alex Darbyshire, Janet Darbyshire, Paul Dargan, Paul Dark, Tace Darling, Kate Darlington, Tom Darton, Guledew Darylile, Manjusha Das, Sukamal Das, Martin Daschel, Joanne Dasgin, Dibyendu Datta, Anna Daunt, Emily Davenport, Mark Davey, Michelle Davey, Miriam Davey, Molly Davey, Arlene David, Mini David, Alexander Davidson, Laura Davidson, Neil Davidson, Richard Davidson, Albert Davies, Alison Davies, Amanda Davies, Amy Davies, Angela Davies, Carolyn Davies, Catrin Davies, Cheryl Davies, Drew Davies, Elaine Davies, Ffyon Davies, Helen Davies, James Davies, Jane Davies, Jeni Davies, Jim Davies, Karen Davies, Kelly Davies, Kim Davies, Louisa Davies, Mark Davies, Matthew Davies, Michelle Davies, Nina Davies, Owen Davies, Patrick Davies, Rachel Davies, Rhys Davies, Ruth Davies, Sarah Davies, Simon Davies, Alison Davis, Gwyneth Davis, Illinos Davis, Julie-Ann Davis, Katherine Davis, Peter Davis, Alexander Davison, Sophia Davison, Mark Davy, Christine Dawe, H Dawe, Mark Dawkins, Danielle Dawson, Elizabeth Dawson, Joy Dawson, Susan Dawson, Tom Dawson, Andrew Daxter, Andrew Day, Helena Day, Jacob Day, Jeremy Day, Lynn Day, Jamie D'Costa, Parijat De, Duneesha de Fonseka, Toni de Freitas, Frederico De Santana Miranda, Eleanor de Sausmarez, Shanika de Silva, Thushan de Silva, Jessica De Sousa, Paulo De Sousa, James de Souza, Anthony de Soyza, Natasha de Vere, Johannes de Vos, Bethan Deacon, Sharon Dealing, Anna Dean, Julie Dean, Katrina Dean, Stephen Dean, Tessa Dean, Jill Deane, James Dear, Effie Dearden, Alison Deary, Catherine Deas, Samuel Debbie, Gabor Debreceni, Vashist Deelchand, Matthew Deeley, Joanne Deery, Emmanuel Defever, Raji Dehulia, Max Deighton, Manuela Del Forno, Arnold Dela Rosa, Lisa Delaney, Amanda Dell, Carrie Demetriou, David DeMets, Jane Democratis, Jacqueline Denham, Emmanuelle Denis, Laura Denley, Craig Denmade, Kathy Dent, Martin Dent, Elise Denton, Tom Denwood, Nishigandh Deole, Darshita Depala, Maria Depante, Randle Derbyshire, Susan Dermody, Amisha Desai, Asmita Desai, Purav Desai, Sanjeev Deshpande, Vai Deshpande, Brendan Devine, Sirjana Devkota, Nicola Dewland, Prakash Dey, Vishal Dey, Rogin Deylami, Jazz Dhaliwal, Kevin Dhaliwal, Jas Dhalliwal, Mandeep Dhanda, Sundip Dhani, Amandeep Dhanoa, Mili Dhar, Devesh Dhasmana, Aman Dhesi, Ekanjali Dhillon, Reiss Dhillon, Pamela Diamond, Priya Dias, Stephanie Diaz, Kayleigh Diaz-Pratt, Debbie Dickerson, Pamela Dicks, Stuart Dickson, Julie Dijo, Sean Dillane, Sarah Diment, Paul Dimitri, Maria Dineen, ThaiHa Dinh, Tri Dinh, Alex Dipper, Laura Dirmantaite, Lisa Ditchfield, Sarah Diver, Lavanya Diwakar, Caroline Dixon, Giles Dixon, Stephen Dixon-Mould, Brice Djeugam, Petr Dlouhy, Laurence Dobbie, Marinela Dobranszky Oroian, Charlotte Dobson, Lee Dobson, Marie Docherty, David Dockrell, James Dodd, Jackie Dodds, Rebecca Dodds, Steve Dodds, Richi Dogra, Erin Doherty, Warren Doherty, Yumiko Doi, Iain Doig, Eleanor Doke, Daniel Dolan, Mark Dolman, Rozzie Dolman, Lisa Donald, Callum Donaldson, Christopher Donaldson, Denise Donaldson, Gillian Donaldson, Kate Donaldson, Joanne Donnachie, Christopher Donnelly, Eilish Donnelly, Ronan Donnelly, Aravindhan Donohoe, Gemma Donohoe, Bryan Donohue, Sinead Donton, Emma Dooks, Grainne Doran, Kane Dorey, Sharon Dorgan, Amanda Dornan, Moonira Dosani, Davinder Dosanjh, Paula Dospinescu, Katie Douglas, Jonathan Douse, Lucy Dowden, Michelle Dower, Kerry Dowling, Sud Dowling, Nicola Downer, Charlotte Downes, Rob Downes, Thomas Downes, Damian Downey, Philippa Downey, Robert Downey, Louise Downs, Simon Dowson, Cornel Dragan, Cristina Dragomir, Cristina Dragos, Maire Drain, Chelsea Drake, Victoria Drew, Olivia Drewett, Celine Driscoll, Helena Drogan, Nikki Drogman, Graham Drummond, Ronald Druyeh, Simon Drysdale, An Du Thinh, Hazel Dube, Judith Dube, Ophias Dube, Stephen Duberley, Hayley Duckles-Leech, Nicola Duff, Emma Duffield, Sam Duff-Miller, Helen Duffy, Lionel Dufour, Annette Duggan, Helen Duggan, Parveen Dugh, Janice Duignan, Mick Duley, Simon Dummer, Andrew Duncan, Christopher Duncan, Fullerton Duncan, Gregory Duncan, Stephanie Dundas, Alessia Dunn, Charlotte Dunn, Damian Dunn, Laura Dunn, Paul Dunn, Charlene Dunne, Karen Dunne, Fiona Dunning, Aidan Dunphy, Venkat Duraiswamy, Beatriz Duran, Ingrid DuRand, Steve Durgacharan, Natalie Duric, Alison Durie, Emily Durie, Hannah Durrington, Haris Duvnjak, Akshay Dwarakanath, Laasya Dwarakanath, Ellen Dwyer, Claudia Dyball, Lee Dyble, Kristyn Dyer, Harvey Dymond, Tom Dymond, Chris Eades, Laura 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Elliott, Toby Elliott, Annie Ellis, Ann-Marie Ellis, Christine Ellis, Kay Ellis, Kaytie Ellis, Tak-Yan Ellis, Yvette Ellis, Megan Ellison, Rahma Elmahdi, Einas Elmahi, Hannah-May Elmasry, Mohammed Elmi, Najla Elndari, Omer Elneima, Mohamed Elokl, Ahmed Elradi, Mohamed Elsaadany, Sally El-Sayeh, Hana El-Sbahi, Tarek Elsefi, Karim El-Shakankery, Robert Elshaw, Hosni El-Taweel, Sarah Elyoussfi, Jonathan Emberey, Jonathan R Emberson, John Emberton, Julian Emmanuel, Ingrid Emmerson, Michael Emms, Florence Emond, Marieke Emonts, Nicu Enachi, Angila Engden, Katy English, Emma Entwistle, Hene Enyi, Marios Erotocritou, Helen Escreet, Peter Eskander, Hanif Esmail, Lise Estcourt, Amy Evans, Brynach Evans, Chris Evans, Daren Evans, Debra Evans, Gail Evans, Gareth Evans, Jennifer Evans, Lisa Evans, Lynn Evans, Margaret Evans, Michelle Evans, Mim Evans, Morgan Evans, Ranoromanana Evans, Ryan Evans, Teriann Evans, Terry John Evans, Tony Eve, Caroline Everden, Serenydd Everden, Hayley Evison, Lynsey Evison, Penny Eyton-Jones, Jacqueline Faccenda, Leila Fahel, Youstina Fahmay, Sara Fairbairn, Terry Fairbairn, Andy Fairclough, Louise Fairlie, Mark Fairweather, Anne Fajardo, Naomi Falcone, Euan Falconer, Jonathan Falconer, John Fallon, Andrea Fallow, David Faluyi, Victoria Fancois, Qayyum Farah, Novin Fard, Amr Farg, Margaret Farinto, Adam Farmer, Katie Farmer, Toni Farmery, Samantha Farnworth, Faiyaz Farook, Hadia Farooq, Sidrah Farooq, Fiona Farquhar, Karen Farrar, Aaron Farrell, Barbara Farrell, James Farthing, Syeda Farzana, Rahmatu Fasina, Azam Fatemi, Mina Fatemi, Nibah Fatimah, Maria Faulkner, Saul N Faust, Joe Fawke, Sinmidele Fawohunre, Abul Fazal, Kelly Feane, Simon Fearby, Alex Feben, Federico Fedel, Daria Fedorova, James Feely-Henderson, Christopher Fegan, Mae Felongco, Lynsey Felton, Tim Felton, Kate Fenlon, Andrea Fenn, Isabelle Fenner, Ciara Fenton, Melisa Fenton, Cameron Ferguson, Jenny Ferguson, Kathryn Ferguson, Katie Ferguson, Stephanie Ferguson, Susan Ferguson, Susie Ferguson, Victoria Ferguson, Denzil Fernandes, Candida Fernandez, Eduardo Fernandez, Maria Fernandez, Sonia Fernandez Lopez, Callum Jeevan Fernando, Ahmed Feroz, Pietro Ferranti, Thais Ferrari Gersogamo, Eleanor Ferrelly, Alexandra Ferrera, Emma Ferriman, Nicholas Fethers, Ben Field, Janet Field, Rebecca Field, Karen Fielder, Lindsey Fieldhouse, Andra Fielding, Julie Fielding, Len Fielding, Sarah Fielding, Asma Fikree, Sarah Ann Filson, Sarah Finbow, Debbie Finch, Joanne Finch, Laurie Finch, Joanne Finden, Natalie Fineman, Lauren Finlayson, Adam Finn, Joanne Finn, Clare Finney, Sofia Fiouni, Jo Fiquet, Dani Fisher, Emily Fisher, James Fisher, Neil Fisher, Meadow Fisher Crisp, Daniel Fishman, Krystofer Fishwick, Lorraine Fitzgerald, Chloe Fitzpatrick-Creamer, Jan Flaherty, Michael Flanagan, Charles Flanders, Cathy Flatters, Julie Fleming, Lucy Fleming, Paul Fleming, William Flesher, Alison Fletcher, Jonathan Fletcher, Lucy Fletcher, Simon Fletcher, Sophie Fletcher, Karen Flewitt, Christopher Flood, Ian Floodgate, Vincent Florence, Sharon Floyd, Kelly Flynn, Rachel Flynn, Sara Flynn, Claire Foden, Adama Fofana, Georgina Fogarty, Claire Foley, Paul Foley, Linda Folkes, Daniela Mock Font, Evodian Fonyonga, Aiwyne Foo, Jane Foo, Andrew Foot, Jayne Foot, Jane Forbes, Jamie Ford, Kathy Ford, Jennifer Foreman, Caroline Fornolles, Adam Forrest, Ellie Forsey, Miranda Forsey, Thomas Forshall, Elliot Forster, Julian Forton, Emily Foster, Joseph Foster, Rachel A Foster, Tracy Foster, Theodora Foukanelli, Angela Foulds, Ian Foulds, Folakemi Fowe, Emily Fowler, Robert Fowler, Stephen Fowler, Caroline Fox, Claire Fox, Daniel Fox, Heather Fox, Jonathan Fox, Lauren Fox, Natalie Fox, Olivia Fox, Simon Fox, Sarah-Jane Foxton, Yasin Fozdar, Rebecca Frake, Alex Francioni, Olesya Francis, Rebecca Francis, Sarah Francis, Theodora Francis-Bacon, Jason Frankcam, Helen Frankland, Gayle Franklin, Jessica Franklin, Darron Franks, Catherine Fraser, Laura Fraser, Sharon Frayling, Martyn Fredlund, Matthew Free, Carol Freeman, Elaine Freeman, Hannah Freeman, Nicola Freeman, Clare Freer, Ian Freestone, Eleanor French, Matthew Frise, Renate Fromson, Claire Froneman, Adam Frosh, John Frost, Victoria Frost, Oliver Froud, Rachel Frowd, Arun Fryatt, Jake Fryer, Janet Fu, Bridget Fuller, Liz Fuller, Neil Fuller, Tracy Fuller, Duncan Fullerton, Jenny Fullthorpe, Carrie Fung, Gayle Fung, Sarah Funnell, John Furness, Charlene Furtado, Andrew Fyfe, Nytianandan G, Elizabeth Gabbitas, Claire Gabriel, Diana Gabriel, Hadiza Gachi, Rose Gad, Joshua Gahir, Sarveen Gajebasia, Katarzyna Gajewska-Knapik, Zacharoula Galani, Christopher Gale, Hugo Gale, Rebecca Gale, Swetha Gali, Karen Galilee, Bernadette Gallagher, Jude Gallagher, Rosie Gallagher, William Gallagher, Joanne Galliford, Catherine Galloway, Chris Galloway, Emma Galloway, Jacqui Galloway, James Galloway, Laura Gamble, Liz Gamble, Brian Gammon, Jaikumar Ganapathi, Ramesh Ganapathy, Kaminiben Gandhi, Sarah Gandhi, Usha Ganesh, Abrar Gani, Emma-James Garden, Antoni D Gardener, Emma Gardiner, Jill Gardiner, Michael Gardiner, Phil Gardiner, Siobhan Gardiner, Caroline Gardiner-Hill, Jonathan Gardner, Mark Garfield, Atul Garg, Nathan Garlick, Justin Garner, Lucie Garner, Zoe Garner, Kimberley Garnett, Robert Garney, Rosaline Garr, Michael Garstka, Peter Gartan, Florence Garty, Rachel Gascoyne, Hyeriju Gashau, Noha Gasmalseed, Michaela Gaspar, Aoife Gatenby, Erin Gaughan, Alok Gaurav, Mariana Gavrila, Jane Gaylard, Emma Gaywood, Catherine Geddie, Alison Geddis, Ian Gedge, Sarah Gee, Minerva Gellamucho, Karzan Gelly, Leila Gelmon, Sandra Gelves-Zapata, Gemma Genato, Susan Gent, Natalie Geoghegan, Chloe George, Sam George, Tina George, Simon Georges, Domonique Georgiou, Peter Gerard, Leigh Gerdes, Louise Germain, Helen Gerrish, Abel Getachew, Louise Gethin, Hisham Ghanayem, Amardeep Ghattaoraya, Anca Gherman, Alison Ghosh, Justin Ghosh, Sudhamay Ghosh, Sarra Giannopoulou, Malick Gibani, Andrew Gibb, Ben Gibbison, Kerry Gibbons, Alex Gibson, Bethan Gibson, Kimberley Gibson, Kirsty Gibson, Sian Gibson, Cat Gilbert, Jeanette Gilbert, Joanne Gilbert, Kayleigh Gilbert, Sean Gilchrist, Benjamin Giles, Mandy Gill, Rose Gill, Lynne Gill, Paul Gillen, Annelies Gillesen, Katherine Gillespie, Matt Gillespie, Elizabeth Gillham, Andrew Gillian, Deborah Gilliland, Robert Gillott, Danielle Gilmour, Kate Gilmour, Theodora Giokanini-Royal, Anna Gipson, Joanna Girling, Rhian Gisby, Angelena Gkioni, Aikaterini Gkoritsa, Effrossyni Gkrania-Klotsas, Amy Gladwell, James Glanville, Jessica Glasgow, Susannah Glasgow, Jon Glass, Lynn Glass, Sharon Glaysher, Lisa Gledhill, Ana Glennon, John Glover, Kyle Glover, Jan Glover Bengtsson, Chevanthy Gnanalingam, Julie Goddard, Wendy Goddard, Emily Godden, Jo Godden, Gillian Godding, Emma Godson, Gerry Gogarty, Sukanya Gogoi, Aiky Goh, Rebeca Goiriz, Sriya Gokaraju, Philip Gold, Raphael Goldacre, Arthur Goldsmith, Portia Goldsmith, Darren Gomersall, Lucia Gomez, Raquel Gomez-Marcos, Ali Gondal, Celia Gonzalez, Jack Goodall, Bob Goodenough, Laura Goodfellow, James Goodlife, Camelia Goodwin, Elizabeth Goodwin, Jayne Goodwin, Paula Goodyear, Rajiv Gooentilleke, Sharif Goolam-Hossen, Michelle Goonasekara, Sheila Gooseman, Shameer Gopal, Peter Gordon, Sally Gordon, Hugh Gorick, Caitlin Gorman, Claire Gorman, Stuart Gormely, Diana Gorog, Jan Gorry, Michelle Gorst, Thomas Gorsuch, Jayshreebahen Gosai, Rebecca Gosling, Sally Gosling, Georgina Gosney, Vanessa Goss, Dzintars Gotham, Naomi Gott, Elizabeth Goudie, Amanda Gould, Angela Gould, Susan Gould, Lysander Gourbault, Anna Gouveia, Abha Govind, Sharon Gowans, Girish Gowda, Rohit Gowda, Pauline Gowdy, Hannah Gower, Thomas Gower, Pankaj Goyal, Sunil Goyal, Sushant Goyal, Beverley Graham, Clive Graham, Jane Graham, Jonathan Graham, Justin Graham, Libby Graham, Sharon Graham, Matthew Graham-Brown, Julia Grahamslaw, Gianluca Grana, Tracyanne Grandison, Louis Grandjean, Alison Grant, Ann Grant, David Grant, 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Griffiths, Yvonne Griffiths, Sofia Grigoriadou, Steph Grigsby, Paul Grist, Stephen Grist, Evelina Grobovaite, Clarissa Grondin, Rachel Groome, Liliana Grosu, Jenny Grounds, Margaret Grout, Helen Grover, Jayne Groves, Neil Grubb, Julie Grundy, Francesca Guarino, Sharada Gudur, Sharazeq Guettari, Shivang Gulati, Vikas Gulia, Pumali Gunasekera, Malin Gunawardena, Kirun Gunganah, Jessica Gunn, Emma Gunter, Alok Gupta, Atul Gupta, Rajeev Gupta, Richa Gupta, Rishi Gupta, Tarun Gupta, Vineet Gupta, Ankur Gupta-Wright, Victoria Guratsky, Alvyda Gureviciute, Sambasivarao Gurram, Bhawana Gurung, Shraddha Gurung, Hazel Guth, Ruth Habibi, Berkin Hack, Pamela Hackney, Christian Hacon, Aiman Haddad, Denise Hadfield, Michalis Hadjiandreou, Nikolaos Hadjisavvas, Leah Hadzik, Anna Haestier, Nauman Hafiz, Rana Hafiz-Ur-Rehman, Javed Hafsa, Samantha Hagan, Jack W Hague, Rosemary Hague, Kate Haigh, Christina Haines, Scott Hainey, Morton Hair, Brigid Hairsine, Juraj Hajnik, Anne Haldeos, Writaja Halder, Jennie Hale, Carmel Halevy, Paul Halford, William Halford, Alistair Hall, Anthony Hall, Claire Hall, Elizabeth Hall, Emma Hall, Fiona Hall, Helen Hall, Jennifer Hall, Kathryn Hall, Bill Hall, Jan Hallas, Kyle Hallas, Charles Hallett, Becky-Lee Halls, Heather Halls, Maryam Hamdollah-Zadeh, Bilal Hameed, Imran Hamid, Mohamad Hamie, Bethany Hamilton, Fergus Hamilton, Leigh Hamilton, Nicola Hamilton, Ruth Hamlin, Eleanor Hamlyn, Beatrice Hammans, Shirley Hammersley, Kate Hammerton, Bev Hammond, Leah Hammond, Sara Hammond, Fiona Hammonds, Ibrahim Hamoodi, Karen Hampshire, Elizabeth Hampson, Jude Hampson, Lucy Hampson, Ozan Hanci, Ian Hancock, Sadiyah Hand, Jasmine Handford, Soran Handrean, Sarah Haney, Sheharyar Hanif, E Hanison, Alison Hanlon, Jennifer Hannah, Amy Hannington, Merhej Hannun, Aidan Hanrath, Anita Hanson, Jane Hanson, Kathryn Hanson, Steve Hanson, Helen Hanwell, Mazhar Ul Haq, Ala Haqiqi, Monjurul Haque, Lesley Harden, Zoe Harding, Simon Hardman, Joanna Hardy, Kumar Haresh, Rachel Harford, Beverley Hargadon, Carolyn Hargreaves, Emily Hargreaves, James Hargreaves, Alice Harin, Mohammed Haris, Edward Harlock, Sandra Harlow, Paula Harman, Tracy Harman, Mark Harmer, Muhammad A Haroon, Charlie Harper, Fiona Harper, Heather Harper, Melanie Harper, Peter Harper, Rosemary Harper, Sarah Harrhy, Sian Harrington, Yasmin Harrington-Davies, Jade Harris, Jess Harris, John Harris, Laura Harris, Marie-Clare Harris, Naomi Harris, Nichola Harris, Sophie Harris, Alex Harrison, David Harrison, Julie Harrison, Laura Harrison, Melanie Harrison, Rowan Harrison, Susie Harrison, Thomas Harrison, Wendy Harrison, Elizabeth Harrod, Ciaran Hart, Dominic Hart, Lisa Hartley, Rosemary Hartley, Ruth Hartley, Tom Hartley, William Hartrey, Phillipa Hartridge, Stuart Hartshorn, Heli Harvala, Alice Harvey, Angela Harvey, Max Harvey, Catherine Harwood, Helen Harwood, Brigitte Haselden, Kazi Hashem, Mohammed Hashimm, Tadaaki Hashimoto, Imranullah Hashmi, Sarah Haskins, Zena Haslam, Adil Hassan, Ali Hassan, Wagae UI Hassan, Waqar Ul Hassan, Sapna Hassasing, Jane Hassell, Philip Hassell, Alex Hastings, Bethany Hastings, Janice Hastings, Stephanie Hatch, Jonathan Hatton, Sheryl Haviland, May Havinden-Williams, Stefan Havlik, Daniel B Hawcutt, Kadean Hawes, Liz Hawes, Nicola Hawes, Annie Hawkins, Catherine Hawkins, Nancy Hawkins, Tanya Hawkins, Dan Hawley, Ed Hawley-Jones, Edward Haworth, Cathy Hay, Amna Hayat, Jamal Hayat, Mohamed-Riyal Hayathu, Tamsin Haydon, Anne Hayes, Jonas Hayes, Kate Hayes, Melony Hayes, Vanessa Hayes, Fiona Hayes, Patrick Hayle, Chloe Haylett, Antara Hayman, Melanie Hayman, Matthew Haynes, Richard Haynes, Rachel Hayre, Sarah Haysom, James Hayward, Patrick Haywood, Tracy Hazelton, Phoebe Hazenberg, Zhengmai He, Elizabeth Headon, Carrie Heal, Brendan Healy, Amy Hearn, Angela Heath, Rowan Heath, Diane Heaton, Kerry Hebbron, Gemma Hector, Andy Hedges, Katrine Hedges, Cheryl Heeley, Elaine Heeney, Rajdeep Heire, Ulla Hemmila, Cassie Hemmings, Scott 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Susie Holt, Alexandra Holyome, Marie Home, Toni Home, Renate Homewood, Kate Hong, Laura Hontoria del Hoyo, Clare Hooper, Sarah Hoosdally, Samantha Hope, Susan Hope, Bridget Hopkins, Peter W Horby, Stephanie Horler, Anil Hormis, Daniel Hornan, Nicola Hornby, Zoey Horne, Latoya Horsford, Megan Horsford, Mark Horsford, Valana Horsham, Alexander Horsley, Ashley Horsley, Elizabeth Horsley, Sarah Horton, Nicola Horton-Turner, Jane Hosea, Toby Hoskins, Muhammad S Hossain, Rashed Hossain, Leanne Hostler, Maxine Hough, Sarah Hough, Brittany Houghton, Catherine Houghton, Iain Houghton, Kathryn Houghton, Rebecca Houlihan, Angela Houston, Hamish Houston, Tawedzegwa Hove, Roseanna Hovvels, Lee How, Laura Howaniec, Laura Howard, Linda Howard, Lucy Howard, Sarah Howard, Stuart Howard, Richard Howard-Griffin, Alison Howarth, Diane Howarth, Serena Howe, Mark Howells, Lyn Howie, Kerry Howlett, Sophie Howlett, Joanne Hoyle, Josh Hrycaiczuk, Naing Zaya Htoon, Su Htwe, Ying Hu, Chiang Ooi Huah Huah, Abby Huckle, Shahzya Huda, Alison Hudak, Lisa Hudig, Alex Hudson, Cara Hudson, Heather Hudson, Peter Hudson, Oli Hudson, Alison Hufton, Connor Huggins, Alistair Hughes, Eithne Hughes, Emma Hughes, Gareth Hughes, Heather Hughes, Luke Hughes, Rachel Hughes, Rebecca Hughes, Samantha Hughes, Stephen Hughes, Vikki Hughes, Wesley Hughes, Lukas Huhn, Ching Hui, Ruth Hulbert, Diana Hull, Grace Hull, Robert Hull, Amanda Hulme, Peter Hulme, Wendy Hulse, George Hulston, Ryan Hum, Laura Humber, Megan Hume, Charlotte Humphrey, Ismay Humphreys, Alasdair Humphries, Joanne Humphries, Lena Hunold, Fiona Hunt, Kristen Hunt, Luke Hunt, Sophie Hunt, Al Hunter, Alexandra Hunter, Isobel Hunter, Karl Hunter, Neil Hunter, George Huntington, Elizabeth Hurditch, Cian Hurley, Katrina Hurley, Mohammed A Husain, Syeda Yusra Husaini, Coralie Huson, Afreen Hussain, Ibraar Hussain, Ifza Hussain, Mohammad Hussain, Muhammad Hussain, Reda Hussain, Sajid Hussain, Samia Hussain, Sanniah Hussain, Wasim Hussain, Yasmin Hussain, Mohammed Hussam El-Din, Raheem Hussein, Rebecca Hussey, Camille Hutchinson, Dorothy Hutchinson, Elizabeth Hutchinson, John Hutchinson, Claire Hutsby, Paula Hutton, Daniella Hydes, Jamie Hyde-Wyatt, Niamh Hynes, Megan Hyslop, Mazen Ibraheim, Abdalla Ibrahim, Ahmed Ibrahim, Asil Ibrahim, Mohamed Ibrahim, Monzeer Ibrahim, Wadah Ibrahim, Adetokunbo I Idowu, Muhammad Idrees, Hina Iftikhar, Mawara Iftikhar, Chukwuemeka Igwe, Mohammad Ijaz, Amaju Ikomi, Clare Iles, Stamatina Iliodromiti, Mary Ilsley, Lorna Ilves, La'ali Imam-Gutierrez, Christopher Imray, Alison Imtiaz, Haider Imtiaz, Claire Ingall, Jack Ingham, Julie Ingham, Rory Ingham, Tejas Ingle, Jennifer Inglis, Anne Ingram, Luke Ingram, Peter Inns, Ken Inweregbu, Andreea A Ionescu, Ana Ionita, Ilian P Iordanov, Anil Ipe, Adil Iqbal, Madiha Iqbal, Mohammed Iqbal, Faisal Iqbal Sait, Jane Ireland, Robert Irons, Mohannad Irshad, Muhammad S Irshad, Janice Irvine, Val Irvine, Pamela Irving, Robert Irving, Mina Ishak, Erica Isherwood, Aminul Islam, Abdurrahman Islim, Ali Ismail, Omar Ismail, Caroline Ison, M'hamedi Israa, Sharon Isralls, Ali Issa, Monica Ivan, Catrina Ivel, Chineze Ivenso, Ashleigh Ivy, Sophie Iwanikiw, Karen Ixer, Menaka Iyer, Mia Iyer, Calum Jack, Amanda Jackson, Anthony Jackson, Ben Jackson, Beth Jackson, Douglas Jackson, Ella Jackson, Hayley Jackson, Helen Jackson, Jane Jackson, Julie Jackson, Karin Jackson, Lauren Jackson, Melanie Jackson, Nicola Jackson, Shane Jackson, Sharon Jackson, Nikita Jacob, Patricia Jacob, Reni Jacob, Nicola Jacques, Terri-Lisa Jacques-Brown, Anisa Jafar, Daniel Jafferji, Ali Jaffery, Chandrashekar Jagadish, Vijay Jagannathan, Sam Jaggard, Mandeep Jagpal, Fernandez R Jaime, Neemisha Jain, Seema Jain, Susan Jain, Sanjay Jaiswal, Danyal Jajbhay, Thomas Jaki, Bintou Jallow, Yusuf Jaly, Sabine Jamal, Zeba Jamal, Yasmin Jameel, Albie James, Christie James, Kate James, Lee James, Linda James, Mark James, Nicholas James, Olivia James, Rebecca James, Ruth James, Tracy James, Jack Jameson, Aaron Jamison, Phoebe Jane, Azara Janmohamed, Sabrina Jansz, Deepa Japp, Lorraine Jappy, Victor Jardim, Catherine Jardine, Emma Jarnell, Ellie Jarvie, Ann Jarvis, Claire Jarvis, Lisa Jarvis, Rosina Jarvis, Patrycja Jastrzebska, Hafsa Javed, Mays Jawad, Lona Jawaheer, Kauky Jawaid, Anu Jayachandran, Dinakaran Jayachandran, Angelina Jayakumar, Deepak Jayaram, Ravi Jayaram, Geeshath Jayasekera, Thilina Jayatilleke, Abi Jayebalan, Saman Jeddi, Vandana Jeebun, Mohammad S Jeelani, Zeynab Jeewa, Emma Jefferson, Katie Jeffery, Helen Jeffrey, Jenni Jeffrey, Rachel Jeffrey, Sue Jeffrey, Nathan Jeffreys, Benjamin Jeffs, Debbie Jegede, Taylor Jemima, Ifan Jenkin, Alison Jenkins, Christopher Jenkins, David Jenkins, Elinor Jenkins, Sarah Jenkins, Sian Jenkins, Stephen Jenkins, Jacqui Jennings, Louise Jennings, Rebecca Jennings, Virginia Jennings, Ellen Jerome, Douglas Jerry, Ellen Jessup-Dunton, Jorge Antonio Jesus Silva, Champa Jetha, Kishan Jethwa, Jeby Jeyachandran, Visuvanathan Jeyakumar, Dharshana Jeyapalan, Shaman Jhanji, Khoo Jian, Zhixin Jiao, Laura Jimenez, Ana Jimenez Gil, Jithin Jith, Teishel Joefield, Navraj Johal, Karine Johannessen, Aisyah Johari, Annie John, Anu John, Navin John, Sarah John, Emma Johns, Margaret Johns, Anne-Marie Johnson, Antoinette Johnson, David Johnson, Emma Johnson, Gillian Johnson, Kathryn Johnson, Katie Johnson, Luke Johnson, Mark Johnson, Nelsonseelan Johnson, Oliver Johnson, Rachel Johnson, Tracy Johnson, Zoe Johnson, Claire Johnston, Janet Johnston, Laura Johnston, Susan Johnston, Victoria Johnston, Dawn Johnstone, Ed Johnstone, Janet Johnstone, Manohar Joishy, Adam Jones, Alistair Jones, Annabel Jones, Ben Jones, Bryony Jones, Carys Jones, Ceri Jones, Charlotte Jones, Christine E Jones, Debra Jones, Emily Jones, Gareth Jones, Geraldine Jones, Hazel Jones, Jac Jones, James Jones, Jamie Jones, Jessica Jones, Jonathon Jones, Julie Jones, Karen Jones, Kate E Jones, Kevin Jones, Laura Jones, Laura M Jones, Lorna Jones, Louise Jones, Mathew Jones, Nicola Jones, Paul Jones, Rhianna Jones, Ruth E Jones, Samantha Jones, Sophie Jones, Stefanie Jones, Steve Jones, Taya Jones, Tim Jones, Tracey Jones, Ramya Jonnalagadda, Rebecca Jordache, Annette Jose, Sanal Jose, Anna Joseph, Joseph Joseph, Rosane Joseph, Sibet Joseph, Dhaara Joshi, Mehul Joshi, Pratichi Joshi, Revati Joshi, Benz Josiah, Tiffany Joyce, Adriel Ju Wen Kwek, Edward Jude, Parminder Judge, Jessica Juhl, Sirisha Jujjavarapu, Mark Juniper, Edmund Juszczak, Deepthi Jyothish, Kasamu Kabiru Dawa, Mark Kacar, Katarina Kacinova, Nikhil Kadam, Rebecca Kahari, Gail Kakoullis, Azad Kala Bhushan, Richard JK Kalayi, Roobala Kaliannan Periyasami, Efthymia Kallistrou, Seika Kalsoom, Elisa Kam, John Kamara, Mohamed Kamara, Ajay Kamath, Prakash Kamath, Ravindra Kamath, Siddharth Arun Kamerkar, Nick Kametas, Musaiwale Kamfose, Arul Kandaswamy, Leia Kane, Osei Kankam, Thogulava Kannan, Abhinav Kant, Vikas Kapil, Ritoo Kapoor, Sonal Kapoor, Sourjya Kar, Janaka Kara, Vasita Kara, Marina Karakantza, Rona Kark, Nicholas Karunaratne, Natashja Kasianczuk, Vidya Kasipandian, Rizwan Kassam, Janarth Kathirgamachelvam, Victoria Katsande, Kulbinder Kaul, Daljit Kaur, Dervinder Kaur, Jasmin Kaur, Jaspreet Kaur, Satvinder Kaur, Zunaira Kausar, Mohammad AA Kawser, Andrea Kay, Sarah Kay, Jossy N Kayappurathu, Callum Kaye, Ahemd Kazeem, Naved Kazi, Sharon Kaznica, Samantha Kearley, Rachel Kearns, Nichola Kearsley, Joanne Keating, John Keating, Liza Keating, Elizabeth Keddie-Gray, Katie Keen, Natalie Keenan, Jonathan Kefas, Stephen Kegg, Laura Keith, Uzoamaka Keke, Tosin Kelani, Joanne Kellett, Jeremy Kellington, Alison Kelly, Conor Kelly, David Kelly, Diane Kelly, Dominic Kelly, Emma Kelly, Laura Kelly, Martin Kelly, Michael Kelly, Rosalind Kelly, Sinead Kelly, Stephen Kelly, Thomas Kelly, Mary Kelly-Baxter, Marketa Keltos, Timothy Kemp, Kelly Kemsley, Alexandra Kendall-Smith, Sarah Kennard, Ann Kennedy, Caroline Kennedy, James Kennedy, Sophie Kennedy-Hay, Julia Kenny, Kelly Kent, Melanie Kent, Lynne Keogan, Alexander Keough, Clement Kerlin, A Kerr, Andrew Kerr, Maria Kerr, Caroline Kerrison, Anthony Kerry, Samantha Kershaw, Helen Kerslake, Ian Kerslake, Helen Kerss, Jocelyn Keshet-Price, Margaret Kevern, Georgina Keyte, Abdul Khadar, Ali Khalid, Muhammad U Khalid, Syed Khalid, Amir Khalil, Asma Khalil, Sijjad Khalil, Abubakar Khan, Ali Khan, Al-Imran Khan, Arham Khan, Asad Khan, Aurangzeb Khan, Burhan Khan, Camran Khan, Fatimah Khan, Kausik Khan, Malik Aamaz Khan, Marria Khan, Mehrunnisha Khan, Mohammad Khan, Mohammed Khan, Nayeem Khan, Omar Khan, Rahe Khan, Rahila Khan, Sabiya Khan, Shabana Khan, Shahul Khan, Shoaib Khan, Tasaduksultan Khan, Waseem Khan, Usman F Khatana, Jibran Khatri, Jyoti Khatri, Hafiza Khatun, Taslima Khatun, Mena Kheia, Jacyntha Khera, Htet Ei Khin, Najaf Khoja, Kiran Khokhar, Jayne Khorsandi, Chloe Khurana, Faith Kibutu, Andrew Kidd, Michelle Kidd, Joe Kidney, Shane Kidney, Will Kieffer, James Kilbane, Caroline Kilby, 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Michelle Lake, Nicki Lakeman, David Lalloo, Fiona Lalloo, Alison Lam, Fiona Lamb, Lucy Lamb, Thomas Lamb, Nick Lambe, Pauline Lambert, Claudia Lameirinhas, Mohammed KG Lami, Abigail Lamikanra, Holly Lamont, Michal Lamparski, Djillali Lamrani, Christine Lanaghan, Rebecca Lanaway, Ivone Lancona-Malcolm, Julia Lancut, Geraldine Landers, Martin J Landray, Matthew Lane, Nicholas Lane, Alidih Lang, Stephen Lang, Daniel Langer, Margaret Langley, Charles Langoya, Emily Langthorne, Taiya Large, Wojciech Lason, Anna Last, Scott Latham, John Latham-Mollart, Afzal Latheef, Darren Latimer, Nang Latt, Carly-Jane Lattimore, Dawn Lau, Eva Lau, Myra Laurenson, Hou Law, Jennifer Law, Jessica Law, Penny Law, Richard Law, Colin Lawler, Mark Lawley, Emma Lawrence, Jo Lawrence, Neil Lawrence, Ryan Lawrie, Jemima Lawson, Joanne Lawson, Louise Lawson, Rebecca Lawton, Michael Lay, Christine Laycock, Reina Layug, Maria Lazo, Vietland Le, Amelia Lea, William Lea, Ian Leadbitter, Thomas Leahy, Richard Lean, Lorna Leandro, Darren Leaning, Sandra Leason, Christina Leaver, Marie Anne Ledingham, Emma Lee, Hannah Lee, Irish Lee, Judith Lee, Sam Lee, Shi Han Lee, Simon Lee, Sindy Lee, Stephanie Lee, Tracey Lee, Xiang Lee, Diana Lees, Jennifer Lees, Helen Legge, Julian Leggett, Katie Leigh-Ellis, Kevan Leighton, Nicky Leitch, Eleni Lekoudis, Petula Lemessy, Nicholas Lemoine, Joana Lemos, Irina Lenchuk, Katy Leng, Katrina Lennon, Liz Lennon, Isabel Lentell, Kelly Leonard, Wen Leong, Nicky Leopold, Oskar Lepiarczyk, Isla Leslie, Eleni Lester, Joe Leung, Ullrich Leuschner, Emma Levell, Chris Levett, Alice Lewin, Michaela Lewin, Alison Lewis, David Lewis, Dee Lewis, Georgina Lewis, Gillian Lewis, Joanne Lewis, Joseph Lewis, Kathryn Lewis, Keir Lewis, Leon Lewis, Lisa Lewis, Marissa Lewis, Rob Lewis, Robert Lewis, Catherine Lewis-Clarke, Lorraine Lewis-Prosser, Katherine Lewiston, Adam Lewszuk, Penny Lewthwaite, Samantha Ley, Anna Li, Jenny Li, Angela Liao, Victoria Licence, David Lieberman, Susan 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Thushika Mahendiran, Siva Mahendran, Sarah Maher, Anistta Maheswaran, Shameera Maheswaran, Tina Maheswaran, Parisa Mahjoob-Afag, Ahmed Mahmood, Farhana Mahmood, Waheed Mahmood, Zahra Mahmood, Hager Mahmoud, Ewan Mahony, Luke Mair, Toluwani Majekdunmi, Kesson Majid, Rupert Major, Jaydip Majumdar, Mohammad KH Majumder, Stephen Makin, Marius Malanca, Hannah Malcolm, Flora Malein, Neeraj Malhan, Ayesha Malik, Gulshan Malik, Mohammed Maljk, Paul Mallett, Petrina Mallinder, Georgia Mallison, Louise Mallon, Edward Malone, Gracie Maloney, Edgar Malundas, Madhu Mamman, Irene Man, Kathy Man, Rossana Mancinelli, Marco Mancuso-Marcello, Tracy Manders, Lauren Manderson, Justin Mandeville, Roope Manhas, Carmen Maniero, Ravi Manikonda, Bobby Mann, Jonathan Manning, Lynne Mannion, Katherine Mansi, Katarina Manso, Dina Mansour, Isheunesu T Mapfunde, Predeesh Mappa, Hemant Maraj, Garikayi Marange, Lisa March, Clare Marchand, Neil Marcus, Maria Marecka, Gomathi Margabanthu, Jordi Margalef, Lavinia Margarit, Georgios Margaritopoulos, Mike Margarson, Fernandez M Maria del Rocio, Teresa Maria Pfyl, Victor Mariano, Helen Maria-Osborn, Ashleigh Maric, Grace Markham, John Markham, Maria Marks, Pamela Marks, Elisabeth Marouzet, Arran Marriott, Cheryl Marriott, Nemonie Marriott, Brian Marsden, Karen Marsden, Paul Marsden, Sarah Marsden, Tracy Marsden, Robyn Marsh, Adam Marshall, Andrew Marshall, Gail Marshall, Henry Marshall, Jaimie Marshall, James Marshall, Jenna Marshall, Nicola Marshall, Riley Marshall, Jennifer Marshall, Samantha Marston, Emmeline Martin, Hayley Martin, Hope Martin, Jane Martin, Karen Martin, Kate Martin, Laila Martin, Michael Martin, Noelia Martin, Tim Martin, Winston Martin, Sarah Martin, Tim Martindale, Marcus Martineau, Alexander Martinez, Lauren Martinez, Jose Carlos Martinez Garrido, Juan Martin-Lazaro, Olivia Martins, Lucas Martins Ferreira, Vijay Kumar Maruthamuthu, Gemma Maryan, Roman Mary-Genetu, Sam Maryosh, Vidan Masani, Diego Maseda, Sheila Mashate, 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Raj, Pradeep Rajagopalan, Nithy Rajaiah, Arvind Rajasekaran, Aylur Rajasri, Thurkka Rajeswaran, Jyothi Rajeswary, Jeyanthy Rajkanna, Gayathri Rajmohan, Ruth Rallan, David Ralphs, Katherine Ralston, Maximilian Ralston, Matsa Ram, Balaji Ramabhadran, Fathima Ramali, Mohamed Ramali, Athimalaipet Ramanan, Shashikira Ramanna, Maheshi Ramasamy, Dhanishta Ramdin, Jozel Ramirez, Mylah Ramirez, Geshwin Ramnarain, Lidia Ramos, Shanthi Ramraj, Alex Ramshaw, Aleem Rana, Ghulam F Rana, Rehman Rana, Abby Rand, James Rand, Helen Randall, Harpal Randheva, Poonam Ranga, Manmeet Rangar, Harini Rangarajan, Sameer Ranjan, Poormina Ranka, Rajesh Rankhelawon, Haley Ranton, Anita Rao, Sandhya Rao, Sanjay Rao, Deepak Rao, Anuja Rasarathnam, Alia Rashid, Khalid Rashid, Simbisai Ratcliff, Sam Ratcliffe, Sophy Ratcliffe, Sanjeev Rath, Mohmad I Rather, Selina Rathore, Aravinden Ratnakumar, Jonathan Ratoff, Deepa Rattehalli, Jason Raw, Hywel Rawlins, Gautam Ray, Michelle Ray, Adam Raymond-White, Dana Raynard, 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Anand Shah, Bhavni Shah, Momin Shah, Neil Shah, Pallav Shah, Priyank Shah, Qasim Shah, Sarfaraz H Shah, Snehal Shah, Suraj Shah, Syed Shah, Wajid Shah, Saarma Shahad, Sousan Shahi, Sipan Shahnazari, Muhammad Shahzeb, Aisha Shaibu, Zara Shaida, Amina Y Shaikh, Maliha Shaikh, Rajit Shail, Mariya Shaji, Muhammad Shakeel, Korah Shalan, Nadia Shamim, Kazi Shams, Alison Shanahan, Thomas Shanahan, Shaminie Shanmugaranjan, Hamed Sharaf, Muhammad Sharafat, Asir Sharif, Ajay Sharma, Akhilesh Sharma, Ash Sharma, Bhawna Sharma, Mona Sharma, Ojasvi Sharma, Poonam Sharma, Rajeev Sharma, Sanjeev Sharma, Sarkhara Sharma, Shriv Sharma, Sonal Sharma, Alexander Sharp, Charles Sharp, Gemma Sharp, Paula Sharratt, Phoebe Sharratt, Katherine Sharrocks, Emma Sharrod, Christopher Shaw, Daisy Shaw, David Shaw, Deborah Shaw, Joanne Shaw, Jonathan Shaw, Lisa Shaw, Tomos G Shaw, Anna Shawcross, Jill Shawe, Lou Shayler, Khuram Shazad, Sophy Shedwell, Jonathan Sheffield, Zak Shehata, Arshiya Sheik, Asif Sheikh, 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Harriet Smith, Hazel Smith, Helen Smith, Jacky Smith, Janice Smith, Jessica Smith, Juliet Smith, Karen Smith, Kate Smith, Kathryn Smith, Katie Smith, Kelly Smith, Kerry Smith, Lara Smith, Linda Smith, Lisa Smith, Loren Smith, Maria Smith, Mel Smith, Oliver Smith, Rachel Smith, Rebecca Smith, Richard Smith, Ruth Smith, Sally Smith, Samantha Smith, Stacey Smith, Stephanie Smith, Susan Smith, Imogen Smith, John Smith, Gemma Smithson, Sue Smolen, Sara Smuts, Naoise Smyth, Annette Snell, David Snell, Luke Snell, Beng So, Michelle Soan, Toluleyi Sobande, Alberto Sobrino Diaz, Basit Sohail, Bina Sohail, Herminder Sohal, Roy Soiza, Mary Sokolowski, Olajumoke Solademi, Krishma Solanki, Babak Soleimani, Amanda Solesbury, Reanne Solly, Louise Solomon, Subash Somalanka, Chandrashekaraiah Somashekar, Raj Sonia, Shiu-Ching Soo, Deepti Sood, Pavandeep Soor, Germanda Soothill, Jennifer Soren, Youssef Sorour, Apina Sothinathan, Pragalathan Sothirajah, Najwa Soussi, Donna Southam, David Southern, Iain 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Holt Walters, James Walters, Jocelyn Walters, Eileen Walton, Lucy Walton, Olivia Walton, Sharon Walton, Susan Walton, Mandy Wan, Thin Wan, Mary Wands, Rachel Wane, Frank Wang, Nick Wang, Ran Wang, Deborah Warbrick, Samantha Warburton, Deborah Ward, Emma Ward, Joanna Ward, Karen Ward, Luke Ward, Nicola Ward, Rachael Ward, Rebecca Ward, Thomas Ward, Tom Ward, Scott A Warden, Adele Wardle, Karen Wardle, Steve Wardle, Hassan Wardy, Scott Waring, Jenny Warmington, Ben Warner, Christian Warner, Lewis Warnock, Sarah Warran, Jade Warren, Lisa Warren, Yolanda Warren, Hannah Warren-Miell, Gill Warwick, Charlotte Washington, Helen Wassall, Hazel J Watchorn, Holly Waterfall, Abby Waters, Donald Waters, Mark Waterstone, Catherine Watkins, Catrin Watkins, Eleanor Watkins, Karen Watkins, Lynn Watkins, Nick Watkins, Abigail Watson, Adam JR Watson, Ekaterina Watson, Eleanor Watson, Paul Watson, Rebecca Watson, Robert Watson, Sandra Watson, Malcolm Watters, Donna Watterson, Daniel Watts, John Watts, Merlin Watts, Victoria Waugh, Emma Wayman, Akhlaq Wazir, Mark Weatherhead, Nick Weatherly, Paul Weaver, Hayley Webb, Kathryn Webb, Kylie Webb, Stephen Webb, Cheryl Websdale, Deborah Webster, Ian Webster, Tim Webster, Kathleen Wedgeworth, Ling Wee, Rebecca Weerakoon, Thanuja Weerasinghe, Janaka Weeratunga, Maria Weetman, Shuying Wei, Immo Weichert, Hugh Welch, James Welch, Leanne Welch, Steven Welch, Samantha Weller, Lucy Wellings, Brian Wells, Susan Wellstead, Berni Welsh, Richard Welsh, Ingeborg Welters, Rachael Welton, Lauren Wentworth, Kate Wesseldine, James Wesson, Jim Wesson, Adam West, Magdelena West, Raha West, Ruth West, Sophie West, Luke Western, Ruth Westhead, Heather Weston, Alice Westwood, Bill Wetherill, Sharon Wheaver, Helen Wheeler, Ben Whelan, Matthew Whelband, Amanda Whileman, Alison Whitcher, Abbie White, Andrew White, Benjamin White, Christopher White, Duncan White, Emily White, James White, Jonathan White, Katie White, Marie White, Nick White, Sarah White, Sonia White, Stephen White, Tracey White, Catherine Whitehead, Anne Whitehouse, Claire Whitehouse, Tony Whitehouse, Julia Whiteley, Sophie Whiteley, Victoria Whiteside, Drew Whitley, Kaitlyn Whitley, Gabriel Whitlingum, David Whitmore, Elizabeth Whittaker, Lindsay Whittam, Andrew Whittingham Hirst, Ashley Whittington, Helen Whittle, Robert Whittle, Suzanne Whyte, Eunice Wiafe, Lou Wiblin, John Widdrington, Jason Wieboldt, Hannah Wieringa, Cornelia Wiesender, Laura Wiffen, Andrew Wight, Christopher Wignall, Danielle Wilcock, Emma Wilcock, Louise Wilcox, Laura Wild, Stephen Wild, Michael Wilde, Peter Wilding, Ritchie Wildman, Tracey Wildsmith, Joe Wileman, Donna Wiles, Joy Wiles, Kate Wiles, Elva Wilhelmsen, Thomas Wiliams, Chloe Wilkes, Janet Wilkie, David Wilkin, Hannah Wilkins, Joy Wilkins, Suzanne Wilkins, Helen Wilkinson, Holly Wilkinson, Iain Wilkinson, Lesley Wilkinson, Martin Wilkinson, Nicola Wilkinson, Sophia Wilkinson, Susan Wilkinson, Tim Wilkinson, Sylvia Willetts, Aimee Williams, Alexandra Williams, Alison Williams, Angharad Williams, Ava Williams, Carl Williams, Caroline V Williams, Claire Williams, Dewi Williams, Gail Williams, Gemma Williams, Gina Williams, Hannah Williams, James Williams, Jayne Williams, Jennie Williams, John Williams, Joseph Williams, Karen Williams, Kathryn Williams, Marie Williams, Matthew Williams, Patricia Williams, Penny Williams, Rachael Williams, Rupert Williams, Samson Williams, Sarah Williams, Sophie Williams, Tamanna Williams, Annie Williamson, Cath Williamson, Catherine Williamson, Dawn Williamson, James D Williamson, Rachel Williamson, Helen Williamson, Bruce Willian, Elizabeth Willis, Emily Willis, Heather Willis, Herika Willis, Joanna Willis, Laura Willmott, Louise Wills, Lucy Willsher, Catherine Willshire, Francesca Willson, Alison Wilson, Andrea Wilson, Antoinette Wilson, Billy Wilson, Catherine Wilson, Eve Wilson, James Wilson, Karen Wilson, Kate Wilson, Lucinda Wilson, Mark Wilson, Matthew Wilson, Toni Wilson, Evie Wiltsher, Marlar Win, Tin Win, Wut Yee Win Win, Lucinda Winckworth, Laura Winder, Piers Winder, Phillip Windrum, Kerry Winham-Whyte, Helen Winmill, Simon Winn, Carmen Winpenny, Helen Winslow, Helen Winter, Jonathan Winter, Pascal Winter, Barbara Winter-Goodwin, Stephen Wisdom, Matthew Wise, Martin Wiselka, Rebecca Wiseman, Sophie Wiseman, Steven Wishart, Holly Wissett, Eric Witele, Nicholas Withers, Janet Wittes, Donna Wixted, Therese Wodehouse, Will Wolf, Nicola Wolff, Kirsten Wolffsohn, Rebecca Wolf-Roberts, Magda Wolna, Elena Wolodimeroff, Adam Wolstencroft, Alan Wong, Charlotte Wong, Chi-Hung Wong, Edwin Wong, Jessica Sue Yi Wong, Kit Y Wong, Lee Wong, Mei Yin Wong, Nick Wong, Sam Wong, Yun Man Wong, Amanda Wood, Caroline Wood, Carrie Anne Wood, Dianne Wood, Fiona Wood, Hannah Wood, Jennifer Wood, Joe Wood, Julia Wood, Kathryn Wood, Lisa Wood, Louise Wood, Michelle Wood, Stephen Wood, Tracy Wood, Ursula Wood, Katharine Woodall, Rebecca Woodfield, Christopher Woodford, Elizabeth Woodford, Jill Woodford, Luke Woodford, Louise Woodhead, Timothy Woodhead, Philip Woodland, Marc Woodman, Debra Woods, Jane Woods, Katherine Woods, Sarah Woods, Zoe Woodward, Rachel Wookey, Megan Woolcock, Gemma Wooldridge, Rebecca Woolf, Chris Woollard, Christopher Woollard, Louisa Woollen, Emma Woolley, Jade Woolley, Daniel Woosey, Dan Wootton, Joanne Wootton, Daniel Worley, Stephy Worton, Jonathan Wraight, Maria Wray, Tim Wreford-Bush, Joanne Wren, Kim Wren, Lynn Wren, Caroline Wrey Brown, Catherine Wright, Demi Wright, Francesca Wright, Imogen Wright, Lee Wright, Lianne Wright, Pete Wright, Rachel Wright, Rebecca Wright, Stephanie Wright, Tim Wright, Caroline Wroe, Hannah Wroe, Henry Wu, Peishan Wu, Pensee Wu, Jonathan Wubetu, Retno Wulandari, Craig Wyatt, Frederick Wyn-Griffiths, Inez Wynter, Bindhu Xavier, Arnold Xhikola, Zhongyang Xia, Huiyuan Xiao, Masseh Yakubi, May Yan, Freda Yang, Yingjia Yang, Michael Yanney, Woei Lin Yap, Nabil Yaqoob, Naairah Yaqub, Salima Yasmin, Bryan Yates, David Yates, Edward Yates, Helen Yates, Julie Yates, Mark Yates, Charlotte Yearwood Martin, Andrew Yeatman, Khin Yein, Fiona Yelnoorkar, Peter Yew, Kawai Yip, Laura Ylquimiche, Laura Ylquimiche Melly, Inez Ynter, H Yong, Jemma Yorke, Jasmine Youens, Abdel Younes Ibrahim, Eoin Young, Gail Young, Louise Young, Richard Young, Asfand Yousafzar, Sajeda Youssouf, Ahmed Yousuf, Chrissie Yu, Bernard Yung, Daniel Yusef, Said Yusef, Intekhab Yusuf, Anna-Sophia Zafar, Silvia Zagalo, Su Zaher, Aqsa Zahoor, Kareem Zaki, Nabhan Zakir, Kasia Zalewska, Ane Zamalloa, Mohsin Zaman, Raisa Zaman, Shakir Zaman, Julie Zamikula, Louise Zammit, Marie Zammit-Mangion, Lynn Zarb, Esther Zebracki, Daniel Zehnder, Lisa Zeidan, Marian Zelman, Xiaobei Zhao, Dongling Zheng, Doreen Zhu, Madiha Zia, Omar Zibdeh, Rabia Zill-E-Huma, Ei Thankt Zin, Veronica Zindonda, Eleanor Zinkin, Vivian Zinyemba, Christos Zipitis, Arkadiusz Zmierczak, Azam Zubir, Roslin Zuha, Naz Zuhra, Rasha Zulaikha, Sabrina Zulfikar, Carol Zullo, Ana Zuriaga-Alvaro, Will Zuurbier, Sheba Zyengi, and University of St Andrews. School of Medicine
Subjects: Male, Convalescent plasma, medicine.medical_treatment, 030204 cardiovascular system & hematology, Antibodies, Viral, Rate ratio, 0302 clinical medicine, RA0421, RA0421 Public health. Hygiene. Preventive Medicine, Medicine, Hospital Mortality, 030212 general & internal medicine, 11 Medical and Health Sciences, Aged, 80 and over, Medicine(all), Mortality rate, Covid19, Articles, 3rd-DAS, General Medicine, Middle Aged, Hospitals, Treatment Outcome, Female, Open label, Coronavirus Infections, Life Sciences & Biomedicine, RM, medicine.medical_specialty, Coronavirus disease 2019 (COVID-19), 03 medical and health sciences, Medicine, General & Internal, SDG 3 - Good Health and Well-being, General & Internal Medicine, Internal medicine, Humans, In patient, Pandemics, COVID-19 Serotherapy, Aged, Mechanical ventilation, Science & Technology, SARS-CoV-2, business.industry, Significant difference, Immunization, Passive, COVID-19, Length of Stay, NIS, R1, Respiration, Artificial, United Kingdom, RM Therapeutics. Pharmacology, RECOVERY Collaborative Group, business
Abstract: Background: Many patients with COVID-19 have been treated with plasma containing anti-SARS-CoV-2 antibodies. We aimed to evaluate the safety and efficacy of convalescent plasma therapy in patients admitted to hospital with COVID-19. Methods: This randomised, controlled, open-label, platform trial (Randomised Evaluation of COVID-19 Therapy [RECOVERY]) is assessing several possible treatments in patients hospitalised with COVID-19 in the UK. The trial is underway at 177 NHS hospitals from across the UK. Eligible and consenting patients were randomly assigned (1:1) to receive either usual care alone (usual care group) or usual care plus high-titre convalescent plasma (convalescent plasma group). The primary outcome was 28-day mortality, analysed on an intention-to-treat basis. The trial is registered with ISRCTN, 50189673, and ClinicalTrials.gov, NCT04381936. Findings: Between May 28, 2020, and Jan 15, 2021, 11558 (71%) of 16287 patients enrolled in RECOVERY were eligible to receive convalescent plasma and were assigned to either the convalescent plasma group or the usual care group. There was no significant difference in 28-day mortality between the two groups: 1399 (24%) of 5795 patients in the convalescent plasma group and 1408 (24%) of 5763 patients in the usual care group died within 28 days (rate ratio 1·00, 95% CI 0·93–1·07; p=0·95). The 28-day mortality rate ratio was similar in all prespecified subgroups of patients, including in those patients without detectable SARS-CoV-2 antibodies at randomisation. Allocation to convalescent plasma had no significant effect on the proportion of patients discharged from hospital within 28 days (3832 [66%] patients in the convalescent plasma group vs 3822 [66%] patients in the usual care group; rate ratio 0·99, 95% CI 0·94–1·03; p=0·57). Among those not on invasive mechanical ventilation at randomisation, there was no significant difference in the proportion of patients meeting the composite endpoint of progression to invasive mechanical ventilation or death (1568 [29%] of 5493 patients in the convalescent plasma group vs 1568 [29%] of 5448 patients in the usual care group; rate ratio 0·99, 95% CI 0·93–1·05; p=0·79). Interpretation: In patients hospitalised with COVID-19, high-titre convalescent plasma did not improve survival or other prespecified clinical outcomes. Funding: UK Research and Innovation (Medical Research Council) and National Institute of Health Research.
Published: 2021
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11. Blood transfusion

Author: Catherine Booth, Shubha Allard, and Susan Robinson
Subjects: General Medicine
Published: 2021
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12. Abstract 153: Identification of a gastrointestinal (GI) toxicity gene signature that predicts GI-toxicity induction in small intestinal organoids

Author: Gary S. Beale, Aude Marine-Bonavita, Valentina Ubertini, Greg Tudor, Francesca Philips, and Catherine Booth
Subjects: Cancer Research, Oncology
Abstract: The early identification of GI toxicity caused by novel therapeutics is crucial for drug development pipelines. GI toxicity is one of the most frequent adverse effects amongst chemotherapeutics. Impaired barrier function and epithelial ulceration can lead to symptoms including diarrhoea, dehydration and susceptibility to systemic infection. We have identified a gene signature that can be utilised in an in vitro organoid model to predict relevant GI toxicity. Methods: Both mice and small intestinal organoids derived from human and mouse were exposed to chemotherapeutic agents known to cause GI toxicity. For in vivo assays mice were treated with 50 or 100mg/kg CPT11 for 4 or 8 hours or 100, 200 or 400mg/kg Iressa for 8 or 24 hours before small intestinal (SI) tissue was removed for RNA-Seq analysis. SI tissue was also analysed by histological staining and damage measured to confirm induction of GI-toxicity. Human and mouse organoids were treated with CPT11 and Iressa for 8 hours at a range of concentrations. The organoids were lysed, RNA extracted and an mRNA-Seq library was generated. Samples were sequenced using 2 × 75bp PE reads on a Next-Seq550. Sequences were aligned to the relevant genomes using BWA and normalised using DESeq2. Normalised gene counts were analysed using Partek Genomics Suite and Ingenuity Pathway Analysis (IPA) was used for gene set enrichment (GSEA) and pathway analysis of differentially expressed genes. Results: For all treatments and time points the mechanism of action (MOA) of each agent was evident from the GSEA and pathway analysis. CPT11 induced DNA damage and cell cycle arrest pathways, whereas Iressa demonstrated downregulation of cell cycle and chromosomal replication and mitotic genes. Analysis of toxicity pathways in IPA identified a number of differentially expressed genes that were consistent with increased toxicity. A gene signature was identified and a multivariate scoring system used to demonstrate signature engagement. This signature was activated in all SI tissues treated with CPT11 or Iressa, irrespective of the MOA of both agents. Conclusions: Deployment of this signature on mouse and human organoids treated with chemotherapeutic agents demonstrated activation of the signature, which could be an alternative method for GI toxicity testing. Further analysis of this signature in gene expression studies from GI toxicity inducing agents submitted to the NCBI Gene expression omnibus (GEO) demonstrated that the signature was activated with a variety of agents with different MOA’s such as NSAIDs, gamma-secretase inhibitors, CDK8/19 inhibitors and CAR-T-cells. Citation Format: Gary S. Beale, Aude Marine-Bonavita, Valentina Ubertini, Greg Tudor, Francesca Philips, Catherine Booth. Identification of a gastrointestinal (GI) toxicity gene signature that predicts GI-toxicity induction in small intestinal organoids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 153.
Published: 2023
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13. Abstract 4350: Pharmacodynamic biomarkers: Evaluation of oncology drug target engagement in human plucked scalp and beard hair

Author: Greg Tudor, Frida Ponthan, Adam Boanas, Aude-Marine Bonavita, and Catherine Booth
Subjects: Cancer Research, Oncology
Abstract: We have developed a method to assess target expression changes following exposure of plucked hairs to chemotherapeutic agents and other potential therapeutics. Successful ex vivo responses provide proof of concept data prior to clinical studies. Plucked hair is a valuable surrogate biomarker tissue to monitor pharmacodynamic (PD) responses in a clinical trial. The collection of hairs is minimally invasive, simple and is amenable to frequent sampling. Since hairs are epithelial appendages, many signaling pathways active in other epithelial tissues, including cancers, are also present in hairs. Highly proliferative plucked human scalp hair can be utilized as a surrogate tissue to measure proliferation, phosphorylation and DNA damage responses after treatment. Furthermore, hairs are highly vascularized, suggesting that administered Test Articles may be delivered efficiently to the hair in a similar manner to other highly vascularized tissues, including many tumor types. Human plucked hair (either from the scalp or beard) was placed in maintenance media in the presence of conventional or targeted chemotherapeutic agents, known for their different mechanisms of action (e.g. Gemcitabine, Carboplatin, Tarceva), for a range of time points. Hairs were then fixed post-exposure and longitudinal sections immunohistochemically labelled for various markers including p-ERK1/2, p-AKT, p-Chk1, g-H2AX, Ki67 and androgen receptor. Quantitative image analysis to measure the level of labelling was performed using an Aperio ScanScope. The relative number of positively labelled cell nuclei, or relative tissue area (depending on the labelling pattern of the target), of the hair outer root sheath labelled was analyzed, along with label intensity. In subsequent trial patients, formaldehyde fixed plucked hairs were returned to the lab, similarly sectioned, labelled and quantified. In addition, where antibody labelling is not possible or more detailed information on the modulation of a pathway may be useful, drug treated plucked hairs can be subjected to Next Generation Sequencing (NGS) gene expression analysis. The expected changes in labelling were observed. For example, Gemcitabine, which is linked to DNA polymerase inhibition, increased p-Chk1 labelling 4-fold after 4-8 hours and strongly induced p53 by 24 hours. Tarceva, an inhibitor of the EGF pathway, decreased levels of both p-ERK1/2 and p-AKT 3-fold after only 10 minutes. The alkylating agent carboplatin increased g-H2AX labelling up to 12-fold after 24 hours. We have demonstrated that plucked hair is a valuable PD biomarker tissue for various chemotherapy agents. Proof of concept studies performed ex vivo can inform the design of clinical validation studies by indicating optimum markers and timepoints. Further, each hair is an independent unit thereby allowing independent replicate tissues (hairs) to be sampled. Citation Format: Greg Tudor, Frida Ponthan, Adam Boanas, Aude-Marine Bonavita, Catherine Booth. Pharmacodynamic biomarkers: Evaluation of oncology drug target engagement in human plucked scalp and beard hair. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4350.
Published: 2023
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14. Evaluation of Plasma Biomarker Utility for the Gastrointestinal Acute Radiation Syndrome in Non-human Primates after Partial Body Irradiation with Minimal Bone Marrow Sparing through Correlation with Tissue and Histological Analyses

Author: Gregory Tudor, Praveen Kumar, Ann M. Farese, Maureen A. Kane, Pengcheng Wang, Thomas J. MacVittie, and Catherine Booth
Subjects: Male, Pathology, medicine.medical_specialty, Epidemiology, Health, Toxicology and Mutagenesis, Radiation Dosage, Article, 030218 nuclear medicine & medical imaging, Jejunum, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Bone Marrow, medicine, Citrulline, Animals, Radiology, Nuclear Medicine and imaging, Gastrointestinal tract, business.industry, Acute Radiation Syndrome, Radiation Exposure, Macaca mulatta, Small intestine, Gastrointestinal Tract, Radiation Injuries, Experimental, Haematopoiesis, medicine.anatomical_structure, chemistry, 030220 oncology & carcinogenesis, Biomarker (medicine), Bone marrow, business, Organ Sparing Treatments, Biomarkers
Abstract: Exposure to total- and partial-body irradiation following a nuclear or radiological incident result in the potentially lethal acute radiation syndromes of the gastrointestinal and hematopoietic systems in a dose-and time-dependent manner. Radiation-induced damage to the gastrointestinal tract is observed within days to weeks post-irradiation. Our objective in this study was to evaluate plasma biomarker utility for the gastrointestinal acute radiation syndrome in non-human primate after partial body irradiation with minimal bone marrow sparing through correlation with tissue and histological analyses. Plasma and jejunum samples from non-human primates exposed to partial body irradiation of 12 Gy with bone marrow sparing of 2.5% were evaluated at various time points from day 0 to day 21 as part of a natural history study. Additionally, longitudinal plasma samples from non-human primates exposed to 10 Gy partial body irradiation with 2.5% bone marrow sparing were evaluated at timepoints out to 180 days post-irradiation. Plasma and jejunum metabolites were quantified via liquid chromatography - tandem mass spectrometry and histological analysis consisted of corrected crypt number, an established metric to assess radiation-induced gastrointestinal damage. A positive correlation of metabolite levels in jejunum and plasma was observed for citrulline, serotonin, acylcarnitine and multiple species of phosphatidylcholines. Citrulline levels also correlated with injury and regeneration of crypts in the small intestine. These results expand the characterization of the natural history of gastrointestinal acute radiation syndrome in non-human primates exposed to partial body irradiation with minimal bone marrow sparing and also provide additional data toward the correlation of citrulline with histological endpoints.
Published: 2020
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15. Proteomic Evaluation of the Natural History of the Acute Radiation Syndrome of the Gastrointestinal Tract in a Non-human Primate Model of Partial-body Irradiation with Minimal Bone Marrow Sparing Includes Dysregulation of the Retinoid Pathway

Author: Amy E. Defnet, Tian Liu, Stephanie Zalesak, Praveen Kumar, Thomas J. MacVittie, Ann M. Farese, Weiliang Huang, Gregory Tudor, Catherine Booth, Maureen A. Kane, and Jianshi Yu
Subjects: Male, Proteome, Epidemiology, medicine.drug_class, Health, Toxicology and Mutagenesis, Retinoic acid, Retinoid X receptor, Radiation Dosage, Article, 030218 nuclear medicine & medical imaging, Retinoids, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Bone Marrow, medicine, Animals, Radiology, Nuclear Medicine and imaging, Retinoid, Gastrointestinal tract, business.industry, Acute-phase protein, Acute Radiation Syndrome, Radiation Exposure, Macaca mulatta, Gastrointestinal Tract, Disease Models, Animal, Radiation Injuries, Experimental, medicine.anatomical_structure, chemistry, 030220 oncology & carcinogenesis, Cancer research, Bone marrow, business, Organ Sparing Treatments, Biomarkers
Abstract: Exposure to ionizing radiation results in injuries of the hematopoietic, gastrointestinal, and respiratory systems, which are the leading causes responsible for morbidity and mortality. Gastrointestinal injury occurs as an acute radiation syndrome. To help inform on the natural history of the radiation-induced injury of the partial body irradiation model, we quantitatively profiled the proteome of jejunum from non-human primates following 12 Gy partial body irradiation with 2.5% bone marrow sparing over a time period of 3 wk. Jejunum was analyzed by liquid chromatography-tandem mass spectrometry, and pathway and gene ontology analysis were performed. A total of 3,245 unique proteins were quantified out of more than 3,700 proteins identified in this study. Also a total of 289 proteins of the quantified proteins showed significant and consistent responses across at least three time points post-irradiation, of which 263 proteins showed strong upregulations while 26 proteins showed downregulations. Bioinformatic analysis suggests significant pathway and upstream regulator perturbations post-high dose irradiation and shed light on underlying mechanisms of radiation damage. Canonical pathways altered by radiation included GP6 signaling pathway, acute phase response signaling, LXR/RXR activation, and intrinsic prothrombin activation pathway. Additionally, we observed dysregulation of proteins of the retinoid pathway and retinoic acid, an active metabolite of vitamin A, as quantified by liquid chromatography-tandem mass spectrometry. Correlation of changes in protein abundance with a well-characterized histological endpoint, corrected crypt number, was used to evaluate biomarker potential. These data further define the natural history of the gastrointestinal acute radiation syndrome in a non-human primate model of partial body irradiation with minimal bone marrow sparing.
Published: 2020
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16. The Natural History of Acute Radiation-induced H-ARS and Concomitant Multi-Organ Injury in the Nonhuman Primate: The MCART experience

Author: Catherine Booth, Kim G. Hankey, Thomas J. MacVittie, Greg L. Tudor, Wanchang Cui, George A. Parker, Eric P. Cohen, and Ann M. Farese
Subjects: medicine.medical_specialty, Epidemiology, business.industry, Health, Toxicology and Mutagenesis, Acute kidney injury, Urology, Acute Radiation Syndrome, Context (language use), Dose-Response Relationship, Radiation, Total body irradiation, medicine.disease, Macaca mulatta, Pathophysiology, Article, Haematopoiesis, medicine.anatomical_structure, Bone Marrow, Concomitant, medicine, Animals, Radiology, Nuclear Medicine and imaging, Bone marrow, business, Whole-Body Irradiation
Abstract: The dose response relationship and corresponding values for mid-lethal dose and slope are used to define the dose- and time-dependent parameters of the hematopoietic acute radiation syndrome. The characteristic time course of mortality, morbidity, and secondary endpoints are well defined. The concomitant comorbidities, potential mortality, and other multi-organ injuries that are similarly dose- and time-dependent are less defined. Determination of the natural history or pathophysiology associated with the lethal hematopoietic acute radiation syndrome is a significant gap in knowledge, especially when considered in the context of a nuclear weapon scenario. In this regard, the exposure is likely ill-defined, heterogenous, and nonuniform. These conditions forecast sparing of bone marrow and increased survival from the acute radiation syndrome consequent to threshold doses for the delayed effects of acute radiation exposure due to marrow sparing, medical management, and use of approved medical countermeasures. The intent herein is to provide a composite natural history of the pathophysiology concomitant with the evolution of the potentially lethal hematopoietic acute radiation syndrome derived from studies that focused on total body irradiation and partial body irradiation with bone marrow sparing. The marked differential in estimated LD50/60 from 7.5 Gy to 10.88 Gy for the total body irradiation and partial body irradiation with 5% bone marrow sparing models, respectively, provided a clear distinction between the attendant multiple organ injury and natural history of the two models that included medical management. Total body irradiation was focused on equivalent LD50/60 exposures. The 10 Gy and 11 Gy partial body with 5% bone marrow sparing exposures bracketed the LD50/60 (10.88 Gy). The incidence, progression, and duration of multiple organ injury was described for each exposure protocol within the hematopoietic acute radiation syndrome. The higher threshold doses for the partial body irradiation with bone marrow sparing protocol induced a marked degree of multiple organ injury to include lethal gastrointestinal acute radiation syndrome, prolonged crypt loss and mucosal damage, immune suppression, acute kidney injury, body weight loss, and added clinical comorbidities that defined a complex timeline of organ injury through the acute hematopoietic acute radiation syndrome. The natural history of the acute radiation syndrome presents a 60-d time segment of multi-organ sequelae that is concomitant with the latent period or time to onset of the evolving multi-organ injury of the delayed effects of acute radiation exposure.
Published: 2021

17. Comparison of PCR versus PCR-Free DNA Library Preparation for Characterising the Human Faecal Virome

Author: George M. Savva, Lesley Hoyles, Andrea Telatin, Evelien M. Adriaenssens, Rebecca Ansorge, Thomas Wileman, Shen-Yuan Hsieh, Mohammad A. Tariq, Simon R. Carding, and Catherine Booth
Subjects: Library preparation, Genome, Viral, Microbiology, Free dna, Article, Bacteriophage, Feces, bacteriophage, Virology, Humans, Human virome, Bacteriophages, Dna viral, Cloning, Molecular, Illumina dye sequencing, Gene Library, Genetics, Genetic diversity, virome, biology, High-Throughput Nucleotide Sequencing, Sequence Analysis, DNA, biology.organism_classification, PCR bias, QR1-502, Gastrointestinal Microbiome, Infectious Diseases, Viruses, Metagenome, Metagenomics, Nucleic Acid Amplification Techniques, Bacteria
Abstract: The human intestinal microbiota is abundant in viruses, comprising mainly bacteriophages, occasionally outnumbering bacteria 10:1 and is termed the virome. Due to their high genetic diversity and the lack of suitable tools and reference databases, the virome remains poorly characterised and is often referred to as “viral dark matter”. However, the choice of sequencing platforms, read lengths and library preparation make study design challenging with respect to the virome. Here we have compared the use of PCR and PCR-free methods for sequence-library construction on the Illumina sequencing platform for characterising the human faecal virome. Viral DNA was extracted from faecal samples of three healthy donors and sequenced. Our analysis shows that most variation was reflecting the individually specific faecal virome. However, we observed differences between PCR and PCR-free library preparation that affected the recovery of low-abundance viral genomes. Using three faecal samples in this study, the PCR library preparation samples led to a loss of lower-abundance vOTUs evident in their PCR-free pairs (vOTUs 128, 6202 and 8364) and decreased the alpha-diversity indices (Chao1 p-value = 0.045 and Simpson p-value = 0.044). Thus, differences between PCR and PCR-free methods are important to consider when investigating “rare” members of the gut virome, with these biases likely negligible when investigating moderately and highly abundant viruses.
Published: 2021

18. Desulfovibrio diazotrophicus sp. nov., a sulfate‐reducing bacterium from the human gut capable of nitrogen fixation

Author: Catherine Booth, Wei Chen, Tianqi Li, Arjan Narbad, Marcelo Bueno Batista, Qixiao Zhai, Lizbeth Sayavedra, and Brandon K. B. Seah
Subjects: Desulfovibrionaceae, Microbiology, 03 medical and health sciences, Nitrogen Fixation, Nitrogenase, Animals, Humans, Phylogeny, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, 0303 health sciences, biology, Bacteria, 030306 microbiology, Sulfates, biology.organism_classification, Isolation (microbiology), Desulfovibrio, Urease complex, Metagenomics, Nitrogen fixation, Oxidation-Reduction
Abstract: Sulfate-reducing bacteria (SRB) are widespread in human guts, yet their expansion has been linked to colonic diseases. We report the isolation, sequencing and physiological characterization of strain QI0027T , a novel SRB species belonging to the class Desulfovibrionia. Metagenomic sequencing of stool samples from 45 Chinese individuals, and comparison with 1690 Desulfovibrionaceae metagenome-assembled genomes recovered from humans of diverse geographic locations, revealed the presence of QI0027T in 22 further individuals. QI0027T encoded nitrogen fixation genes and based on the acetylene reduction assay, actively fixed nitrogen. Transcriptomics revealed that QI0027T overexpressed 42 genes in nitrogen-limiting conditions compared to cultures supplemented with ammonia, including genes encoding nitrogenases, a urea uptake system and the urease complex. Reanalyses of 835 public stool metatranscriptomes showed that nitrogenase genes from Desulfovibrio bacteria were expressed in six samples suggesting that nitrogen fixation might be active in the gut environment. Although frequently thought of as a nutrient-rich environment, nitrogen fixation can occur in the human gut. Animals are often nitrogen limited and have evolved diverse strategies to capture biologically active nitrogen, ranging from amino acid transporters to stable associations with beneficial microbes that provide fixed nitrogen. QI0027T is the first Desulfovibrio human isolate for which nitrogen fixation has been demonstrated, suggesting that some sulfate-reducing bacteria could also play a role in the availability of nitrogen in the gut.
Published: 2022
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19. Multiparametric MRI of early tumor response to immune checkpoint blockade in metastatic melanoma

Author: Mary A. McLean, Catherine Booth, Lee Brown, Pippa Corrie, Ilse Patterson, Amy Frary, Michal Sulikowski, Joshua D. Kaggie, Doreen Lau, Ferdia A. Gallagher, Frank Riemer, Francis Scott, Luigi Aloj, Doreen Milne, Andrew B. Gill, Andrew N. Priest, Bruno Carmo, Martin J. Graves, Kevin M. Brindle, Jean-Martin Lapointe, Arthur Lewis, Lau, Doreen [0000-0002-7623-2401], McLean, Mary A [0000-0002-3752-0179], Priest, Andrew N [0000-0002-9771-4290], Gill, Andrew B [0000-0002-9287-9563], Lapointe, Jean-Martin [0000-0003-0141-4603], Corrie, Pippa G [0000-0003-4875-7021], Gallagher, Ferdia A [0000-0003-4784-5230], and Apollo - University of Cambridge Repository
Subjects: Oncology, Male, CTLA-4 antigen, Cancer Research, medicine.medical_specialty, tumor, Metastatic melanoma, medicine.medical_treatment, Immunology, Text mining, Vascularity, Internal medicine, Immunotherapy Biomarkers, melanoma, Immunology and Allergy, Medicine, Humans, Multiparametric Magnetic Resonance Imaging, Prospective cohort study, Immune Checkpoint Inhibitors, RC254-282, Aged, Pharmacology, business.industry, Melanoma, Immunity, Multiparametric MRI, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, biomarkers, Immunotherapy, Middle Aged, medicine.disease, Immune checkpoint, Blockade, Response Evaluation Criteria in Solid Tumors, translational medical research, Molecular Medicine, Female, immunotherapy, medicine.symptom, business
Abstract: BackgroundImmune checkpoint inhibitors are now standard of care treatment for many cancers. Treatment failure in metastatic melanoma is often due to tumor heterogeneity, which is not easily captured by conventional CT or tumor biopsy. The aim of this prospective study was to investigate early microstructural and functional changes within melanoma metastases following immune checkpoint blockade using multiparametric MRI.MethodsFifteen treatment-naïve metastatic melanoma patients (total 27 measurable target lesions) were imaged at baseline and following 3 and 12 weeks of treatment on immune checkpoint inhibitors using: T2-weighted imaging, diffusion kurtosis imaging, and dynamic contrast-enhanced MRI. Treatment timepoint changes in tumor cellularity, vascularity, and heterogeneity within individual metastases were evaluated and correlated to the clinical outcome in each patient based on Response Evaluation Criteria in Solid Tumors V.1.1 at 1 year.ResultsDifferential tumor growth kinetics in response to immune checkpoint blockade were measured in individual metastases within the same patient, demonstrating significant intertumoral heterogeneity in some patients. Early detection of tumor cell death or cell loss measured by a significant increase in the apparent diffusivity (Dapp) (papp), was consistently higher in the pseudoprogressive and true progressive lesions, compared with the responding lesions throughout the first 12 weeks of treatment. These preceded tumor regression and significant tumor vascularity changes (Ktrans, ve, and vp) detected after 12 weeks of immunotherapy (pConclusionsMultiparametric MRI demonstrated potential for early detection of successful response to immune checkpoint inhibitors in metastatic melanoma.
Published: 2021

20. Targeted Metabolomics Reveals Metabolomic Signatures Correlating Gastrointestinal Tissue to Plasma in a Mouse Total-body Irradiation Model

Author: Fei Li, Maureen A. Kane, Ann M. Farese, Gregory Tudor, Catherine Booth, Jace W. Jones, Zachary Clifford, and Thomas J. MacVittie
Subjects: Male, Epidemiology, Health, Toxicology and Mutagenesis, Metabolite, Pharmacology, Article, 030218 nuclear medicine & medical imaging, Mice, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Metabolomics, medicine, Animals, Radiology, Nuclear Medicine and imaging, chemistry.chemical_classification, Acute Radiation Syndrome, Total body irradiation, Small intestine, Amino acid, Gastrointestinal Tract, Mice, Inbred C57BL, Radiation Injuries, Experimental, Jejunum, medicine.anatomical_structure, chemistry, 030220 oncology & carcinogenesis, Experimental pathology, Biomarkers, Whole-Body Irradiation, Targeted metabolomics
Abstract: High-throughput, targeted metabolomics was used to identify early time point small intestine and plasma metabolite markers of gastrointestinal acute radiation syndrome. The small intestine metabolite markers were cross-correlated to plasma metabolites in order to identify minimally invasive circulating markers. The radiation exposure covered lethal and sub-lethal gastrointestinal acute radiation syndrome. The small intestine and plasma metabolite profiles were generated at 1 and 3 days post-exposure following total-body irradiation. The small intestine and plasma metabolite profiles for mice receiving radiation at day 1 and 3 post-exposure were significantly different from sham-irradiated mice. There were 14 metabolite markers identified at day 1 and 18 metabolite markers at day 3 that were small intestine-specific plasma markers of gastrointestinal acute radiation syndrome. A number of the identified metabolites at day 1 were amino acids. Dysregulation of amino acid metabolism at 24-hours post total-body irradiation provides potential insight into the initial inflammatory response during gastrointestinal acute radiation syndrome.
Published: 2019
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21. Proteomic Evaluation of the Acute Radiation Syndrome of the Gastrointestinal Tract in a Murine Total-body Irradiation Model

Author: Keely Pierzchalski, Catherine Booth, Maureen A. Kane, Gregory Tudor, Jianshi Yu, Weiliang Huang, Claire L. Carter, Thomas J. MacVittie, and Jace W. Jones
Subjects: Male, Proteomics, Epidemiology, Health, Toxicology and Mutagenesis, Retinoic acid, Inflammation, Biology, Article, 030218 nuclear medicine & medical imaging, Mice, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Downregulation and upregulation, Ileum, Tandem Mass Spectrometry, medicine, Animals, Radiology, Nuclear Medicine and imaging, Gastrointestinal tract, Proteomic Profiling, Acute Radiation Syndrome, Dose-Response Relationship, Radiation, Total body irradiation, Gastrointestinal Tract, Mice, Inbred C57BL, Radiation Injuries, Experimental, chemistry, 030220 oncology & carcinogenesis, Cancer research, medicine.symptom, Biomarkers, Whole-Body Irradiation, Chromatography, Liquid
Abstract: Radiation exposure to the gastrointestinal system contributes to the acute radiation syndrome in a dose- and time-dependent manner. Molecular mechanisms that lead to the gastrointestinal acute radiation syndrome remain incompletely understood. Using a murine model of total-body irradiation, C57BL/6J male mice were irradiated at 8, 10, 12, and 14 Gy and assayed at day 1, 3, and 6 after exposure and compared to nonirradiated (sham) controls. Tryptic digests of gastrointestinal tissues (upper ileum) were analyzed by liquid chromatography-tandem mass spectrometry on a Waters nanoLC coupled to a Thermo Scientific Q Exactive hybrid quadrupole-orbitrap mass spectrometer. Pathway and gene ontology analysis were performed with Qiagen Ingenuity, Panther GO, and DAVID databases. A number of trends were identified in our proteomic data including pronounced protein changes as well as protein changes that were consistently up regulated or down regulated at all time points and dose levels interrogated. Time- and dose-dependent protein changes, canonical pathways affected by irradiation, and changes in proteins that serve as upstream regulators were also identified. Additionally, proteins involved in key processes including inflammation, radiation, and retinoic acid signaling were identified. The proteomic profiling conducted here represents an untargeted systems biology approach to identify acute molecular events that will be useful for a greater understanding of animal models and may be potentially useful toward the development of medical countermeasures and/or biomarkers.
Published: 2019
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22. Characterizing the Natural History of Acute Radiation Syndrome of the Gastrointestinal Tract: Combining High Mass and Spatial Resolution Using MALDI-FTICR-MSI

Author: Catherine Booth, Jace W. Jones, Kim G. Hankey, Thomas J. MacVittie, Ann M. Farese, Maureen A. Kane, Gregory Tudor, George A. Parker, and Claire L. Carter
Subjects: Male, Pathology, medicine.medical_specialty, Epidemiology, Health, Toxicology and Mutagenesis, Cell, Article, Mass spectrometry imaging, 030218 nuclear medicine & medical imaging, 03 medical and health sciences, 0302 clinical medicine, medicine, Animals, Radiology, Nuclear Medicine and imaging, Intestinal Mucosa, Receptor, Epithelial cell differentiation, Gastrointestinal tract, Lamina propria, Chemistry, Acute Radiation Syndrome, Lipid Metabolism, Macaca mulatta, Epithelium, Mitochondria, Gastrointestinal Tract, Jejunum, medicine.anatomical_structure, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, 030220 oncology & carcinogenesis, Biomarkers
Abstract: The acute radiation syndrome of the gastrointestinal tract has been histologically characterized, but the molecular and functional mechanisms that lead to these cellular alterations remain enigmatic. Mass spectrometry imaging is the only technique that enables the simultaneous detection and cellular or regional localization of hundreds of biomolecules in a single experiment. This current study utilized matrix-assisted laser desorption/ionization mass spectrometry imaging for the molecular characterization of the first natural history study of gastrointestinal acute radiation syndrome in the nonhuman primate. Jejunum samples were collected at days 4, 8, 11, 15, and 21 following 12-Gy partial-body irradiation with 2.5% bone marrow sparing. Mass spectrometry imaging investigations identified alterations in lipid species that further understanding of the functional alterations that occur over time in the different cellular regions of the jejunum following exposure to high doses of irradiation. Alterations in phosphatidylinositol species informed on dysfunctional epithelial cell differentiation and maturation. Differences in glycosphingolipids of the villi epithelium that would influence the absorptive capacity and functional structure of the brush border membrane were detected. Dichotomous alterations in cardiolipins indicated altered structural and functional integrity of mitochondria. Phosphatidylglycerol species, known regulators of toll-like receptors, were detected and localized to regions in the lamina propria that contained distinct immune cell populations. These results provide molecular insight that can inform on injury mechanism in a nonhuman primate model of the acute radiation syndrome of the gastrointestinal tract. Findings may contribute to the identification of therapeutic targets and the development of new medical countermeasures.
Published: 2019
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23. The Gastrointestinal Subsyndrome of the Acute Radiation Syndrome in Rhesus Macaques: A Systematic Review of the Lethal Dose-response Relationship With and Without Medical Management

Author: Ann M. Farese, Catherine Booth, Christopher S Potten, George A. Parker, Thomas J. MacVittie, Kim G. Hankey, Gregory Tudor, and William E. Jackson
Subjects: Gastrointestinal Diseases, Epidemiology, Health, Toxicology and Mutagenesis, Lung injury, Bioinformatics, Article, 030218 nuclear medicine & medical imaging, 03 medical and health sciences, 0302 clinical medicine, Animals, Medicine, Radiology, Nuclear Medicine and imaging, biology, business.industry, Lethal dose, Acute Radiation Syndrome, Dose-Response Relationship, Radiation, biology.organism_classification, Macaca mulatta, Gastrointestinal Tract, Natural history, Dose–response relationship, Rhesus macaque, 030220 oncology & carcinogenesis, Time course, business, Dose rate
Abstract: Well-characterized animal models that mimic the human response to potentially lethal doses of radiation are required to assess the efficacy of medical countermeasures under the criteria of the Food and Drug Administration “animal rule”. Development of a model for the acute GI-ARS requires knowledge of the radiation dose response relationship and time course of mortality and morbidity across the acute and prolonged gastrointestinal radiation syndrome. The nonhuman primate, Rhesus macaque, is a relevant animal model that has been used to determine the efficacy of medical countermeasures to mitigate major signs of morbidity and mortality relative to the H-, GI-ARS and lung injury. It can be used to assess the natural history of GI damage, concurrent multiple organ injury and aspects of the mechanism of action for acute radiation exposure and treatment. A systematic review of relevant studies that determined the dose response relationship for the gastrointestinal acute and prolonged radiation syndrome in the Rhesus macaque relative to radiation dose, quality, dose rate, exposure uniformity and use of medical management has never been performed. The selection of data cohorts was made from the following sources: Ovid Medline (1957-present), PubMed (1954-present), AGRICOLA (1976-present), Web of Science (1954-present and US HHS REPORT (2002 to present). The following terms were used: Rhesus, total-body irradiation, total-body x-irradiation, TBI, irradiation, gamma radiation, gastrointestinal system, LD50, LD50/60, hematopoietic radiation syndrome, Macaca mulatta, whole-body irradiation, nonhuman primate, NHP, monkey, primates, gastrointestinal radiation syndrome, mortality, and nuclear radiation. The reference lists of all studies, published and unpublished, were reviewed for additional studies. The total number of hits across all search sites was 3,923. Thirty-two (32) studies, utilizing 1,364 total nonhuman primates (NHP), Rhesus Macaca mulatta, were evaluated to provide an informative and consistent review. Ten (10) primary studies provided adequate data to establish dose response relationships (DRR)s for the acute GI-ARS. The DRRs were determined for uniform or non-uniform total-body irradiation (TBI) with 240–250kVp or 2Mev x-radiation, Co-60 gamma radiation, isotope-derived gamma-radiation, reactor- and nuclear weapon-derived mixed gamma:neutron radiation and 6MV LINAC-derived photons delivered at various dose rates from a total-body, bilateral, rotational or unilateral exposure aspect in the absence or presence of medical management. The DRRs established by a probit analysis versus linear dose relationship were characterized by two main parameters or dependent variables, a slope and LD50/15. Respective LD50/15 values for the primary studies without use of medical management that used 250kVp x-radiation (three studies, total n=234), 2Mev x-radiation (n=84) and Co-60 gamma-radiation (n=90) were: 250kVp x-radiation (605rad [572, 636], 581rad [542, 621], and 608rad [369, 871]); Co-60 gamma -radiation (708rad [683, 736]) and 2Mev x-radiation (671rad [632, 715]). The respective slopes were steep and ranged from 0.402 to 1.915 probits per linear dose. The DRR, LD50/15 value and slope were also determined for total-body, non-uniform, unilateral, pulse-rate exposure to reactor-derived mixed gamma:neutron radiation (total n=111). The LD50/15 value was 482rad with a slope of 0.992 [0.142, 1.99]. Two contemporary studies provided DRRs for NHP exposed to TBI or PBI with 5% bone marrow sparing (PBI/BM5) with 6 MV LINAC-derived photons with administration of medical management. The LD50/15 and slopes were: TBI, 11.33 Gy [10.81, 11.75] with a slope of 0.917 [0.701, 1.133]; PBI/BM5, 12.01 Gy [11.71, 12.52] with a slope of 1.001 [0.599, 1.415]. The PBI/BM5 model also provided a modified, longitudinal, image-based histopathology of damage to the small intestine through 180d post exposure relative to normal, un-irradiated age-matched tissue. Secondary studies (12) provided data sets of appropriate limited studies that did not describe a DRR for the GI-ARS but included adequate control data sets or a range of mortality values from high-dose range exposure in the conduct of establishing the DRR for the H-ARS, were used to support the threshold and mid- to high-lethal dose range for the GI-ARS. Tertiary studies (12) provided data sets that supported the incidence, time course, severity, histology, physiology and biomarkers of radiation-induced GI injury in the NHP. The acute GI-ARS and prolonged GI injury were evaluated at threshold doses to include initial concomitant GI injury consequent to TBI at mid- to high-dose lethal H-ARS. The available evidence provided a relatively consistent and reliable data base that characterized the DRR and survival probability over the 15 day time course for the acute GI-ARS in young Rhesus macaques exposed to total-body and partial-body irradiation with several sources of differing radiation quality, dose rate and non-uniform exposure with and without medical management. The GI-ARS occurred with concomitant evolution of the H-ARS. The image-based histopathology provided a clear demonstration of the pronounced radiation-induced damage to the GI system along the longitudinal course from acute to prolonged damage through the 180d study duration. The DRRs for the GI-ARS are characterized by steep slopes and relative LD50/15 values that reflected the biological effect of radiation quality, exposure aspect, dose rate and medical management over a range in time from 1954–2015.
Published: 2019
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24. Histopathological Features of the Development of Intestine and Mesenteric Lymph Node Injury in a Nonhuman Primate Model of Partial-body Irradiation with Minimal Bone Marrow Sparing

Author: Ann M. Farese, Kyle Takayama, Gregory Tudor, Catherine Booth, George A. Parker, Na Li, and Thomas J. MacVittie
Subjects: Male, Pathology, medicine.medical_specialty, Colon, Epidemiology, Health, Toxicology and Mutagenesis, H&E stain, Article, 030218 nuclear medicine & medical imaging, Jejunum, 03 medical and health sciences, 0302 clinical medicine, Bone Marrow, Submucosa, medicine, Animals, Mesenteric lymph nodes, Mesentery, Radiology, Nuclear Medicine and imaging, Lymph node, business.industry, Macaca mulatta, Intestines, Radiation Injuries, Experimental, medicine.anatomical_structure, 030220 oncology & carcinogenesis, Lymph Nodes, Lymph, Bone marrow, business
Abstract: Male rhesus macaques were subjected to partial-body irradiation at 10, 11, or 12 Gy with 5% bone marrow protection. Animals were euthanized when dictated by prospectively determined clinical parameters, or at approximately 180 days following irradiation. Histological sections of jejunum, colon and mesenteric lymph node were stained with hematoxylin and eosin as well as a battery of histochemical and immunohistochemical stains. The immediate post-irradiation histopathological alterations in the jejunum and colon were based primarily on injury to rapidly proliferating crypt epithelial cells, though there was evidence of additional radiation-induced fibrogenic responses. There was substantial resolution of the radiation-related mucosal injury through the observation period, but microscopically visible defects in mucosal structure persisted to the end of the observation period. In the later stages of the observation period the jejunum and colon had overt fibrosis that was most commonly located in the submucosa and serosa, with less microscopically discernible involvement of the mucosa. Mesenteric lymph nodes had an immediate post-irradiation reduction in cellularity due to the known effects of irradiation on lymphoid cell populations. In later stages of the observation period the lymph nodes also developed fibrotic changes, possibly related to transmigration of immunomodulatory cells and/or signaling molecules from the radiation-damaged intestine.
Published: 2019
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25. Discovery of a crystalline sulforaphane analog with good solid-state stability and engagement of the Nrf2 pathway in vitro and in vivo

Author: Shuping Dong, Yolanda Sanchez, Tindy Li, Claudia E. Murar, Anthony J. Wilson, Brent W. Mccleland, Hongwei Qi, Patricia L. Podolin, Marc Galop, Christopher J. Moody, Hongxing Yan, John Yonchuk, Roderick S. Davis, Alan P. Graves, Catherine Booth-Genthe, Joseph P. Foley, Jen-Pyng Kou, Jeffrey K. Kerns, Brian Bolognese, James F. Callahan, Mary Mentzer, Lawrence Wolfe, Jeffrey C. Boehm, and Robin Carr
Subjects: Antioxidant, NF-E2-Related Factor 2, medicine.medical_treatment, Clinical Biochemistry, Gene Expression, Pharmaceutical Science, medicine.disease_cause, Biochemistry, Antioxidants, Cell Line, Structure-Activity Relationship, chemistry.chemical_compound, Isothiocyanates, Thiocarbamates, In vivo, Drug Discovery, medicine, Animals, Humans, Thermal stability, Molecular Biology, Kelch-Like ECH-Associated Protein 1, Natural product, Molecular Structure, Organic Chemistry, KEAP1, Rats, Mice, Inbred C57BL, Oxidative Stress, Solubility, chemistry, Sulfoxides, Isothiocyanate, Biophysics, Molecular Medicine, Cyclobutanes, Heme Oxygenase-1, Oxidative stress, Signal Transduction, Sulforaphane
Abstract: The antioxidant natural product sulforaphane (SFN) is an oil with poor aqueous and thermal stability. Recent work with SFN has sought to optimize methods of formulation for oral and topical administration. Herein we report the design of new analogs of SFN with the goal of improving stability and drug-like properties. Lead compounds were selected based on potency in a cellular screen and physicochemical properties. Among these, 12 had good aqueous solubility, permeability and long-term solid-state stability at 23 °C. Compound 12 also displayed comparable or better efficacy in cellular assays relative to SFN and had in vivo activity in a mouse cigarette smoke challenge model of acute oxidative stress.
Published: 2019
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26. Salvage of refractory post-transfusion hyperhaemolysis by targeting hyperinflammation and macrophage activation with tocilizumab

Author: Nay Win, Banu Kaya, Filipa Barroso, Catherine Booth, Sarah Bennett, Paul Telfer, and Frederick Chen
Subjects: business.industry, Post transfusion, Transfusion Reaction, Hematology, Macrophage Activation, Antibodies, Monoclonal, Humanized, Hemolysis, chemistry.chemical_compound, Tocilizumab, Refractory, chemistry, Immunology, Macrophage, Medicine, Humans, business
Published: 2021

27. Impact of mush reservoir processes on the source, frequency and composition of large-scale volcanic eruptions

Author: Alison C Rust, R. S. J. Sparks, Catherine Booth, and Matthew D. Jackson
Subjects: geography, geography.geographical_feature_category, Volcano, Scale (ratio), Earth science, Geology
Published: 2021
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28. Extracellular vesicles released by the human gut symbiont Bacteroides thetaiotaomicron in the mouse intestine are enriched in a selected range of proteins that influence host cell physiology and metabolism

Author: Udo Wegmann, Andrew J. Goldson, Simon R. Carding, Maria Guirro, Catherine Booth, Emily Jones, Kathryn Cross, Ian R. Brown, Dezső Módos, Arlaine Brion, Avani Ranjit, Patrick Gunning, Régis Stentz, Padhmanand Sudhakar, William A. Bryant, Ariadna Miquel-Clopés, and Tamas Korcsmaros
Subjects: Human gut, Chemistry, Metabolism, Bacteroides thetaiotaomicron, Extracellular vesicles, Mouse Intestine, Cell biology
Abstract: It is becoming increasingly clear that bacterial extracellular vesicles (BEVs) produced by members of the intestinal microbiota can contribute to microbe-host cell interactions that impact on host health. A major unresolved question is the nature of the cargo packaged into these BEVs and how they can impact on host cell function. Here we have analysed the proteome of BEVs produced by the major human gut symbiont Bacteroides thetaiotaomicron in both in vitro cultures using defined and complex medias, and in vivo in fed or fasted animals to determine the impact of nutrient stress on the BEV proteome, and to identify proteins specifically enriched in BEVs produced in vivo. In contrast to BEVs produced in vitro where limiting nutrient provision resulted in an increase in a large fraction of proteins, the protein content of BEVs extracted from fasted versus fed mice was less affected with similar numbers of proteins showing increased and decreased abundance. We identified 102 proteins exclusively enriched in BEVs in vivo of which the majority (66/102) were enriched independently of their expression in the parent cells implicating the existence of an active mechanism to drive the selection of a group of proteins for their secretion into BEVs within the intestine. Amongst these abundantly expressed proteins in BEVs in vivo were a bile salt hydrolase and a dipeptidyl peptidase IV that were characterised further and shown to be active and able to degrade host-derived substrates with defined roles in metabolism. Collectively these findings provide additional evidence for the role of BEVs in microbiota-host interactions with their contents playing key roles in the maintenance of intestinal homeostasis, and host metabolism.
Published: 2020
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29. Establishing Pediatric Mouse Models of the Hematopoietic Acute Radiation Syndrome and the Delayed Effects of Acute Radiation Exposure

Author: Hailin Feng, P. Artur Plett, Hui Lin Chua, Catherine Booth, Gregory Tudor, Carol H. Sampson, Alexa Fisher, Rajendran Sellamuthu, Andrea M. Patterson, Thomas J. MacVittie, Barry P. Katz, Christie M. Orschell, Steven J. Miller, and Sasidhar Vemula
Subjects: Filgrastim, Hematopoietic System, Biophysics, Physiology, Pediatrics, Radiation Tolerance, 030218 nuclear medicine & medical imaging, Polyethylene Glycols, 03 medical and health sciences, Mice, 0302 clinical medicine, Radioresistance, medicine, Animals, Humans, Radiology, Nuclear Medicine and imaging, Increased blood urea nitrogen, Acute radiation exposure, Radiation, business.industry, Acute Radiation Syndrome, Haematopoiesis, Disease Models, Animal, 030220 oncology & carcinogenesis, medicine.symptom, business, Weight gain, Pegfilgrastim, Radiation response, Whole-Body Irradiation, medicine.drug
Abstract: Medical countermeasures (MCMs) for hematopoietic acute radiation syndrome (H-ARS) should be evaluated in well-characterized animal models, with consideration of at-risk populations such as pediatrics. We have developed pediatric mouse models of H-ARS and delayed effects of acute radiation exposure (DEARE) for efficacy testing of MCMs against radiation. Male and female C57BL/6J mice aged 3, 4, 5, 6, 7 and 8 weeks old (±1 day) were characterized for baseline hematopoietic and gastrointestinal parameters, radiation response, efficacy of a known MCM, and DEARE at six and 12 months after total-body irradiation (TBI). Weanlings (age 3 weeks) were the most radiosensitive age group with an estimated LD50/30 of 712 cGy, while mice aged 4 to 8 weeks were more radioresistant with an estimated LD50/30 of 767-787 cGy. Female weanlings were more radiosensitive than males at 3 and 4 weeks old but became significantly more radioresistant after the pubertal age of 5 weeks. The most dramatic increase in body weight, RBC counts and intestinal circumference length occurred from 3 to 5 weeks of age. The established radiomitigator Neulasta® (pegfilgrastim) significantly increased 30-day survival in all age groups, validating these models for MCM efficacy testing. Analyses of DEARE among pediatric survivors revealed depressed weight gain in males six months post-TBI, and increased blood urea nitrogen at 12 months post-TBI which was more severe in females. Hematopoietic DEARE at six months post-TBI appeared to be less severe in survivors from the 3- and 4-week-old groups but was equally severe in all age groups by 12 months of age. Similar to our other acute radiation mouse models, there was no appreciable effect of Neulasta used as an H-ARS MCM on the severity of DEARE. In summary, these data characterize a pediatric mouse model useful for assessing the efficacy of MCMs against ARS and DEARE in children.
Published: 2020

30. 673 Precision microbiome mapping identifies a microbiome signature predictive of Immune checkpoint inhibitor response across multiple research study cohorts

Author: Matthew R. Robinson, Doreen Milne, David H. Adams, Christine Parkinson, Catherine Booth, Pippa Corrie, David Bruce, Sarah Welesh, Kevin Vervier, Simon R. Harris, Trevor D. Lawley, and Emily Barker
Subjects: 0301 basic medicine, Shotgun sequencing, Melanoma, medicine.medical_treatment, Computational biology, Immunotherapy, Biology, medicine.disease, Genome, Immune checkpoint, 03 medical and health sciences, 030104 developmental biology, 0302 clinical medicine, Metagenomics, 030220 oncology & carcinogenesis, medicine, Microbiome, Reference genome
Abstract: Background The gut microbiome of cancer patients appears to be associated with response to Immune Checkpoint Inhibitor (ICIs) treatment.1–4 However, the bacteria linked to response differ between published studies. Methods Longitudinal stool samples were collected from 69 patients with advanced melanoma receiving approved ICIs in the Cambridge (UK) MELRESIST study. Pretreatment samples were analysed by Microbiotica, using shotgun metagenomic sequencing. Microbiotica’s sequencing platform comprises the world’s leading Reference Genome Database and advanced Microbiome Bioinformatics to give the most comprehensive and precise mapping of the gut microbiome. This has enabled us to identify gut bacteria associated with ICI response missed using public reference genomes. Published microbiome studies in advanced melanoma,1–3renal cell carcinoma (RCC) and non-small cell lung cancer (NSCLC)4 were reanalysed with the same platform. Results Analysis of the MELRESIST samples showed an overall change in the microbiome composition between advanced melanoma patients and a panel of healthy donor samples, but not between patients who subsequently responded or did not respond to ICIs. However, we did identify a discrete microbiome signature which correlated with response. This signature predicted response with an accuracy of 93% in the MELRESIST cohort, but was less predictive in the published melanoma cohorts.1–3 Therefore, we developed a bioinformatic analytical model, incorporating an interactive random forest model and the MELRESIST dataset, to identify a microbiome signature which was consistent across all published melanoma studies. This model was validated three times by accurately predicting the outcome of an independent cohort. A final microbiome signature was defined using the validated model on MELRESIST and the three published melanoma cohorts. This was very accurate at predicting response in all four studies combined (91%), or individually (82–100%). This signature was also predictive of response in a NSCLC study and to a lesser extent in RCC. The core of this signature is nine bacteria significantly increased in abundance in responders. Conclusions Analysis of the MELRESIST study samples, precision microbiome profiling by the Microbiotica Platform and a validated bioinformatic analysis, have enabled us to identify a unique microbiome signature predictive of response to ICI therapy in four independent melanoma studies. This removes the challenge to the field of different bacteria apparently being associated with response in different studies, and could represent a new microbiome biomarker with clinical application. Nine core bacteria may be driving response and hold potential for co-therapy with ICIs. Ethics Approval The study was approved by Newcastle & North Tyneside 2 Research Ethics Committee, approval number 11/NE/0312. References Matson V, Fessler J, Bao R, et al. The commensal microbiome is associated with anti-PD-1 efficacy in metastatic melanoma patients. Science 2018;359(6371):104–108. Gopalakrishnan V, Spencer CN, Nezi L, et al. Gut microbiome modulates response to anti-PD-1 immunotherapy in melanoma patients. Science 2018;359(6371):97–103. Frankel AE, Coughlin LA, Kim J, et al. Metagenomic shotgun sequencing and unbiased metabolomic profiling identify specific human gut microbiota and metabolites associated with immune checkpoint therapy efficacy in melanoma patients. Neoplasia 2017;19(10):848–855. Routy B, Le Chatelier E, Derosa L, et al. Gut microbiome influences efficacy of PD-1-based immunotherapy against epithelial tumors. Science 2018;359(6371):91–97.
Published: 2020
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31. Desulfovibrio diazotrophica sp. nov., a sulphate reducing bacterium from the human gut capable of nitrogen fixation

Author: Qixiao Zhai, M. Bueno Batista, Tianqi Li, Lizbeth Sayavedra, Wei Chen, Arjan Narbad, Brandon K. B. Seah, and Catherine Booth
Subjects: Clostridia, biology, Metagenomics, Nitrogen fixation, Nitrogenase, Diazotroph, Desulfovibrionaceae, biology.organism_classification, Desulfovibrio, Bacteria, Microbiology
Abstract: Sulphate-reducing bacteria (SRB) are widespread in human guts, yet their expansion has been linked to colonic diseases. We report the isolation, genome sequencing, and physiological characterisation of a novel SRB species belonging to the class Deltaproteobacteria (QI0027T). Phylogenomic analysis revealed that the QI0027T strain belongs to the genus Desulfovibrio with its closest relative being Desulfovibrio legallii. Metagenomic sequencing of stool samples from 45 individuals, as well as comparison with 1690 Desulfovibrionaceae metagenome-assembled genomes, revealed the presence of QI0027T in at least 22 further individuals. QI0027T encoded nitrogen fixation genes and based on the acetylene reduction assay, actively fixed nitrogen. Transcriptomics revealed that QI0027T overexpressed 45 genes in nitrogen limiting conditions as compared to cultures supplemented with ammonia, including nitrogenases, an urea uptake system and the urease enzyme complex. To the best of our knowledge, this is the first Desulfovibrio human isolate for which nitrogen fixation has been demonstrated. This isolate was named Desulfovibrio diazotrophica sp. nov., referring to its ability to fix nitrogen (‘diazotroph’).ImportanceAnimals are often nitrogen limited and have evolved diverse strategies to capture biologically active nitrogen. These strategies range from amino acid transporters to stable associations with beneficial microbes that can provide fixed nitrogen. Although frequently thought as a nutrient-rich environment, nitrogen fixation can occur in the human gut of some populations, but so far it has been attributed mainly to Clostridia and Klebsiella based on sequencing. We have cultivated a novel Desulfovibrio from human gut origin which encoded, expressed and actively used nitrogen fixation genes, suggesting that some sulphate reducing bacteria could also play a role in the availability of nitrogen in the gut.
Published: 2020
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32. Positive direct antiglobulin tests in patients with COVID-19

Author: Catherine Booth, Sean Platton, Laura E. Green, Kurtis Lee, Julia Lancut, Fiona Regan, and Nicole Mendes
Subjects: Adult, 2019-20 coronavirus outbreak, Letter, Coronavirus disease 2019 (COVID-19), business.industry, Reverse Transcriptase Polymerase Chain Reaction, SARS-CoV-2, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Immunology, MEDLINE, COVID-19, Hematology, Middle Aged, Antibodies, Viral, Virology, Young Adult, Immunology and Allergy, Medicine, Humans, In patient, Letters, business, Aged
Published: 2020

33. Bifidobacterium breve UCC2003 Exopolysaccharide Modulates the Early Life Microbiota by Acting as a Potential Dietary Substrate

Author: Deborah Püngel, Ian J. Colquhoun, Jennifer Ketskemety, Melissa A. E. Lawson, Shabhonam Caim, Lindsay J. Hall, Douwe van Sinderen, Tamas Korcsmaros, Matthew J. Dalby, Agatha Treveil, and Catherine Booth
Subjects: 0301 basic medicine, Metabolite, 030106 microbiology, Mutant, ved/biology.organism_classification_rank.species, Microbial metabolism, lcsh:TX341-641, digestive system, Article, Microbiology, Transcriptome, 03 medical and health sciences, chemistry.chemical_compound, Metabolomics, fluids and secretions, 16S rRNA profiling, early life, Microbiome, model colon, cross-feeding, Bifidobacterium, Nutrition and Dietetics, Bifidobacterium breve, biology, ved/biology, exopolysaccharides, biology.organism_classification, metabolomics, 030104 developmental biology, chemistry, diet, lcsh:Nutrition. Foods and food supply, Food Science
Abstract: Background: Bifidobacterium represents an important early life microbiota member. Specific bifidobacterial components, exopolysaccharides (EPS), positively modulate host responses, with purified EPS also suggested to impact microbe&ndash, microbe interactions by acting as a nutrient substrate. Thus, we determined the longitudinal effects of bifidobacterial EPS on microbial communities and metabolite profiles using an infant model colon system. Methods: Differential gene expression and growth characteristics were determined for each strain, Bifidobacterium breve UCC2003 and corresponding isogenic EPS-deletion mutant (B. breve UCC2003del). Model colon vessels were inoculated with B. breve and microbiome dynamics monitored using 16S rRNA sequencing and metabolomics (NMR). Results: Transcriptomics of EPS mutant vs. B. breve UCC2003 highlighted discrete differential gene expression (e.g., eps biosynthetic cluster), though overall growth dynamics between strains were unaffected. The EPS-positive vessel had significant shifts in microbiome and metabolite profiles until study end (405 h), with increases of Tyzzerella and Faecalibacterium, and short-chain fatty acids, with further correlations between taxa and metabolites which were not observed within the EPS-negative vessel. Conclusions: These data indicate that B. breve UCC2003 EPS is potentially metabolized by infant microbiota members, leading to differential microbial metabolism and altered metabolite by-products. Overall, these findings may allow development of EPS-specific strategies to promote infant health.
Published: 2020
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34. The Uptake, Trafficking, and Biodistribution of Bacteroides thetaiotaomicron Generated Outer Membrane Vesicles

Author: Régis Stentz, Aimee Parker, Sonia Fonseca, Simon R. Carding, Ulrike Mayer, Kathryn Cross, Emily J. Jones, Thomas Wileman, Isabelle Hautefort, Catherine Booth, and Ariadna Miquel-Clopés
Subjects: Microbiology (medical), lcsh:QR1-502, Gut flora, Endocytosis, Microbiology, lcsh:Microbiology, 03 medical and health sciences, bacterial extracellular vesicles, 030304 developmental biology, GI-tract, 0303 health sciences, biology, gut microbiota, 030306 microbiology, Chemistry, biology.organism_classification, In vitro, Microvesicles, Cell biology, Bacteroides thetaiotaomicron, Paracellular transport, Bacterial outer membrane, microvesicles, Intracellular, outer membrane vesicles
Abstract: Gram-negative bacteria ubiquitously produce and release nano-size, non-replicative outer membrane vesicles (OMVs). In the gastrointestinal (GI-) tract, OMVs generated by members of the intestinal microbiota are believed to contribute to maintaining the intestinal microbial ecosystem and mediating bacteria–host interactions, including the delivery of bacterial effector molecules to host cells to modulate their physiology. Bacterial OMVs have also been found in the bloodstream although their origin and fate are unclear. Here we have investigated the interactions between OMVs produced by the major human gut commensal bacterium, Bacteroides thetaiotaomicron (Bt), with cells of the GI-tract. Using a combination of in vitro culture systems including intestinal epithelial organoids and in vivo imaging we show that intestinal epithelial cells principally acquire Bt OMVs via dynamin-dependent endocytosis followed by intracellular trafficking to LAMP-1 expressing endo-lysosomal vesicles and co-localization with the perinuclear membrane. We observed that Bt OMVs can also transmigrate through epithelial cells via a paracellular route with in vivo imaging demonstrating that within hours of oral administration Bt OMVs can be detected in systemic tissues and in particular, the liver. Our findings raise the intriguing possibility that OMVs may act as a long-distance microbiota–host communication system.
Published: 2020
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35. Abstract P074: MB097: A therapeutic consortium of bacteria clinically-defined by precision microbiome profiling of immune checkpoint inhibitor patients with potent anti-tumor efficacy in vitro and in vivo

Author: Matthew J. Robinson, Kevin Vervier, Simon Harris, Amy Popple, Dominika Klisko, Robyne Hudson, Ghaith Bakdash, Laure Castan, Clelia Villemin, David J. Adams, Doreen Milne, Catherine Booth, Christine Parkinson, Roy Rabbie, Sarah J. Welsh, Emily Barker, Katie Dalchau, Pippa Corrie, and Trevor Lawley
Subjects: Cancer Research, Immunology
Abstract: Independent groups have demonstrated that the pre-treatment gut microbiome of cancer patients impacts the subsequent response to Immune Checkpoint Inhibitor (ICIs) therapy [1-4]. However, each study identified different sets of bacteria linked to outcome, which has limited the development of drug response biomarkers and clinic-first design of novel microbiome-based therapeutics. The Cambridge (UK) MELRESIST study includes a cohort of advanced melanoma patients receiving approved ICIs. Pre-treatment stool samples from MELRESIST were analysed by Microbiotica using shotgun metagenomic sequencing. Microbiotica's platform comprises the leading Reference Genome Database to give the most comprehensive and precise mapping of the gut microbiome. A bioinformatic analysis identify a small discrete microbiome signature that was different between responders and non-responders. We extended this signature by reanalysing three published melanoma cohorts [1-3] using the Microbiotica platform. The resultant bacterial signature predicted whether or not a patient responded to anti-PD1-based therapy with an accuracy of 91% in all four studies combined and was also an effective biomarker for each cohort individually. We validated the signature using a NSCLC study [4] indicating that it has great potential as a clinical biomarker for a number of indications. The signature was strongly skewed towards species raised in abundance in responding patients, suggesting that the microbiome influences ICI treatment primarily through bacteria that enhance the efficacy of the drugs. At the core of the signature was nine species strongly associated a positive outcome, which we hypothesized to be a central driver of drug response. MB097 is a consortium comprised of all nine bacteria. In a syngeneic mouse model of cancer, MB097 was able inhibit tumor growth, but most strikingly was potently synergistic when dose with anti-PD1. To understand the mechanisms by which these bacteria drive an anti-tumor response, we have profiled the bacteria individually and as a consortium in multiple assays with primary human immune cells. The bacteria strongly activate dendritic cells with a number inducing high levels of IL-12 relative to IL-10. These bacteria-stimulated dendritic cells went on to trigger Cytotoxic T Lymphocytes (CTLs) to upregulate Granzyme B, Perforin and IFNg. Further, we have demonstrated that these primed CTLs are very effective at tumor cell killing in vitro. In summary, Microbiotica's precision microbiome profiling and the MELRESIST study has allowed us to identify a consortium of bacteria, MB097, strongly linked to response in multiple melanoma cohorts and a NSCLC study. The consortium drives immune-mediated tumor killing in vivo and in vitro. MB097 is being scaled up for manufacture as a novel co-therapy with ICIs. References 1 Matson V et al Science (2018) 359:104 2 Gopalakrishnan V Science (2018) 359:97 3 Frankel AE et al Neoplasia (2017) 19:848 4 Routy B et al Science (2018) 359:91 Citation Format: Matthew J. Robinson, Kevin Vervier, Simon Harris, Amy Popple, Dominika Klisko, Robyne Hudson, Ghaith Bakdash, Laure Castan, Clelia Villemin, David J. Adams, Doreen Milne, Catherine Booth, Christine Parkinson, Roy Rabbie, Sarah J. Welsh, Emily Barker, Katie Dalchau, Pippa Corrie, Trevor Lawley. MB097: A therapeutic consortium of bacteria clinically-defined by precision microbiome profiling of immune checkpoint inhibitor patients with potent anti-tumor efficacy in vitro and in vivo [abstract]. In: Abstracts: AACR Virtual Special Conference: Tumor Immunology and Immunotherapy; 2021 Oct 5-6. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(1 Suppl):Abstract nr P074.
Published: 2022
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36. Bifidobacterium breveUCC2003 exopolysaccharide modulates the early life microbiota by acting as a dietary substrate

Author: Ian J. Colquhoun, Shabhonam Caim, Deborah Püngel, Douwe van Sinderen, Jennifer Ketskemety, Catherine Booth, Lindsay J. Hall, Matthew J. Dalby, Agatha Treveil, Melissa A. E. Lawson, and Tamas Korcsmaros
Subjects: 0303 health sciences, Bifidobacterium breve, biology, 030306 microbiology, ved/biology, Metabolite, Mutant, ved/biology.organism_classification_rank.species, Microbial metabolism, biology.organism_classification, Microbiology, Transcriptome, 03 medical and health sciences, chemistry.chemical_compound, Metabolomics, chemistry, Escherichia, Microbiome, 030304 developmental biology
Abstract: Members of the genusBifidobacteriumrepresent an important bacterial group for promoting health during early life. Previous studies have indicated that bifidobacterial exopolysaccharides (EPS) are involved in host interactions, with purified EPS also suggested to modulate microbe-microbe interactions by acting as a nutrient substrate. To further explore the role of EPS as a potential dietary component, we determined the longitudinal effects of bifidobacterial EPS on microbial communities and metabolite profiles using an infant model colon system.Bifidobacterium breveUCC2003 was utilised as a representative early life bifidobacterial strain, and a corresponding isogenic EPS-deletion mutant (B. breveUCC2003 EPS-). Initial transcriptomics analysis of the EPS mutant vs. parentB. breveUCC2003 strain highlighted differential expression in a discrete number of genes, including theepsbiosynthetic cluster, though overall growth dynamics between the two strains were unaffected. Model colon vessels were inoculated withB. brevestrains and microbiome dynamics were monitored using metataxonomic (via 16S rRNA sequencing) and metabolomic (via1H NMR) approaches. Baseline early life microbiota profiles were similar between vessels, with persistence ofB. breve(EPS+ and EPS-) observed between 0-36h. Within the EPS-positive vessel there was a significant shift in microbiome and metabolite profiles until the end of the study (405h); we observed increases ofEscherichiaandTyzzerella, and short-chain fatty acids including acetate, propionate and formate, including further correlations between taxa and metabolites which were not observed in the EPS-negative vessel. These data indicate that theB. breveUCC2003 EPS is potentially being metabolised by members of the infant microbial community, leading to differential microbial metabolism and altered metabolite by-products. Overall, these findings may allow for development of EPS-specific strategies to beneficially alter the early life microbiota to promote infant health.
Published: 2019
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37. Quantification of membrane transporter proteins in human lung and immortalized cell lines using targeted quantitative proteomic analysis by isotope dilution nanoLC–MS/MS

Author: Philip C. Smith, John K. Fallon, Nicole Houvig, and Catherine Booth-Genthe
Subjects: Adult, Male, Proteomics, 0301 basic medicine, Adolescent, Clinical Biochemistry, Quantitative proteomics, Indicator Dilution Techniques, Pharmaceutical Science, 030226 pharmacology & pharmacy, Madin Darby Canine Kidney Cells, Xenobiotics, Analytical Chemistry, Young Adult, 03 medical and health sciences, Dogs, 0302 clinical medicine, Isotopes, Tandem Mass Spectrometry, Cell Line, Tumor, Drug Discovery, Gene expression, Lysis buffer, Animals, Humans, Lung, Chromatography, High Pressure Liquid, Spectroscopy, Differential centrifugation, PROSTAGLANDIN TRANSPORTER, Chemistry, Membrane Transport Proteins, Biological Transport, Transporter, 030104 developmental biology, Biochemistry, Female, Homogenization (biology)
Abstract: Information is needed on the expression of transporters in lung to inform drug development and therapeutic decisions. Much of the information currently available is from semiquantitative gene expression or immunometric densitometry studies reported in the literature. NanoLC–MS/MS (MRM mode) isotope dilution targeted quantitative proteomics was used here to quantify twelve selected transporters in fresh human lung membrane fraction samples and in the membrane fraction of selected immortalized human lung epithelial cell line samples. Fractionation was undertaken by homogenization in crude membrane lysis buffer followed by differential centrifugation of the homogenate. In lung membranes we found OATPs to be the most highly expressed transporters of those measured, followed by PEPT2 and ABCs (P-gp & BCRP). SLC22A transporters (OCTs 2 & 3 and OCTN1) were also found to be expressed. OATP2A1, also known as the prostaglandin transporter, was the most highly expressed transporter, being low in two subjects who were at least occasional smokers. One subject, a non-smoker, had an OATP2A1 concentration that was 8.4 times higher than the next nearest concentration, which itself was higher than the concentration of any other transporter. OATP2A1 is known, from gene expression and animal functional studies, to be present in lung. These results inform the understanding of xenobiotic disposition in the lung and show the distinct profile of transporters in lung compared to other tissues.
Published: 2018
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38. Abstract 1783: Precision microbiome profiling identifies a novel biomarker predictive of Immune Checkpoint Inhibitor response in multiple cohorts and a potent therapeutic consortium of bacteria

Author: Pippa Corrie, Sarah J. Welsh, Christine Parkinson, Catherine Booth, Matthew J. Robinson, Simon R. Harris, Trevor D. Lawley, Emily Barker, Roy Rabbie, David Bruce, Kevin Vervier, David H. Adams, and Doreen Milne
Subjects: Oncology, Cancer Research, medicine.medical_specialty, biology, Melanoma, Cancer, medicine.disease, Immune system, Metagenomics, Internal medicine, medicine, biology.protein, Biomarker (medicine), Microbiome, Antibody, Reference genome
Abstract: Four independent international groups have demonstrated that the pre-treatment gut microbiome of cancer patients is associated with the subsequent response to treatment with Immune Checkpoint Inhibitors (ICI) [1-4]. However, each study identified different bacteria as being linked to outcome, which has limited the development of drug response biomarkers and novel microbiome-based therapeutics. Here we describe the identification of a microbial signature predictive of response to ICI across multiple melanoma studies, and a derived Live Bacterial Therapeutic with potent anti-tumour activity. MELRESIST is a single centre, prospective melanoma patient data and biosample collection research study. We collected longitudinal stool samples from 69 patients with advanced melanoma who received standard anti-PD-1+/- anti-CTLA-4 antibodies. Shotgun metagenomic sequencing analysis of the baseline stool microbiome was done using Microbiotica's platform, which comprises the world's leading Reference Genome Database to give the most comprehensive and precise mapping of gut microbiomes. Using 6 months progression-free survival as our cut-off for response, the analysis revealed a small but discrete microbiome signature that differentiated responders and non-responders with an accuracy of 93%. We extended this signature by reanalysing another 3 melanoma patient stool sample sequence datasets [1-3] using the Microbiotica platform, and a machine learning-based bioinformatic model. The resultant bacterial signature accurately predicted response when all 4 studies when combined (91%), as well as when the cohorts were analysed individually (82-100%). We validated the model using independent cohorts and the signature using NSCLC and Renal Cell Carcinoma (RCC) datasets [4]. The latter indicated the bacteria associated with response may differ slightly between indications. At the core of the signature was 9 bacteria that were all overrepresented in patients that responded to ICI treatment. Notably as a consortium, these 9 bacteria demonstrated tumor growth inhibition when dosed in a syngeneic mouse model. These strains also stimulate primary immune cells in vitro leading to tumor cell killing. In summary, we have identified a microbiome biomarker that is predictive of response to ICI treatment in multiple clinical studies from different countries. In addition, a unique set of bacteria derived from the signature has great therapeutic potential in combination with ICIs. References 1 Matson V et al Science (2018) 359:104 2 Gopalakrishnan V Science (2018) 359:97 3 Frankel AE et al Neoplasia (2017) 19:848 4 Routy B et al Science (2018) 359:91 Citation Format: Matthew J. Robinson, Kevin Vervier, Simon Harris, Roy Rabbie, Doreen Milne, Catherine Booth, Christine Parkinson, Sarah J. Welsh, David Bruce, Emily Barker, David Adams, Pippa Corrie, Trevor D. Lawley. Precision microbiome profiling identifies a novel biomarker predictive of Immune Checkpoint Inhibitor response in multiple cohorts and a potent therapeutic consortium of bacteria [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1783.
Published: 2021
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39. Blood transfusion

Author: Catherine Booth and Shubha Allard
Subjects: General Medicine
Published: 2017
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40. The Uptake, Trafficking, and Biodistribution of

Author: Emily J, Jones, Catherine, Booth, Sonia, Fonseca, Aimee, Parker, Kathryn, Cross, Ariadna, Miquel-Clopés, Isabelle, Hautefort, Ulrike, Mayer, Tom, Wileman, Régis, Stentz, and Simon R, Carding
Subjects: Bacteroides thetaiotaomicron, gut microbiota, bacterial extracellular vesicles, Microbiology, microvesicles, biodistribution, outer membrane vesicles, organoid monolayer, Original Research, GI-tract
Abstract: Gram-negative bacteria ubiquitously produce and release nano-size, non-replicative outer membrane vesicles (OMVs). In the gastrointestinal (GI-) tract, OMVs generated by members of the intestinal microbiota are believed to contribute to maintaining the intestinal microbial ecosystem and mediating bacteria–host interactions, including the delivery of bacterial effector molecules to host cells to modulate their physiology. Bacterial OMVs have also been found in the bloodstream although their origin and fate are unclear. Here we have investigated the interactions between OMVs produced by the major human gut commensal bacterium, Bacteroides thetaiotaomicron (Bt), with cells of the GI-tract. Using a combination of in vitro culture systems including intestinal epithelial organoids and in vivo imaging we show that intestinal epithelial cells principally acquire Bt OMVs via dynamin-dependent endocytosis followed by intracellular trafficking to LAMP-1 expressing endo-lysosomal vesicles and co-localization with the perinuclear membrane. We observed that Bt OMVs can also transmigrate through epithelial cells via a paracellular route with in vivo imaging demonstrating that within hours of oral administration Bt OMVs can be detected in systemic tissues and in particular, the liver. Our findings raise the intriguing possibility that OMVs may act as a long-distance microbiota–host communication system.
Published: 2019

41. Differential expression of soluble receptor for advanced glycation end-products (sRAGE) in mice susceptible or resistant to chronic colitis

Author: Sheena M. Cruickshank, John McLaughlin, James Wilson, Ajanta Chakraborty, Larisa Logunova, Namshik Han, Catherine Booth, Michael Bramhall, Andy Brass, and Kevin Rich
Subjects: biology, business.industry, Inflammation, medicine.disease, biology.organism_classification, Trichuris muris, RAGE (receptor), Immune system, Glycation, Immunology, medicine, Colitis, medicine.symptom, Receptor, business, Pathological
Abstract: AimsIdentifying the factors that contribute to chronicity in inflamed colitic tissue is not trivial. However, in mouse models of colitis, we can investigate at preclinical timepoints. We sought to validate murine Trichuris muris infection as a model for identification of factors that promote development of chronic colitis.MethodsWe compared preclinical changes in mice with a resolving immune response to T. muris (resistant) versus mice that fail to expel the worms and develop chronic colitis (susceptible). Findings were then validated in healthy controls and patients with suspected or confirmed IBD.ResultsThe Receptor for Advanced Glycation End Products (Rage) was highly dysregulated between resistant and susceptible mice prior to the onset of any pathological signs. Increased soluble RAGE (sRAGE) in the serum and faeces of resistant mice correlated with reduced colitis scores. Mouse model findings were validated in a preliminary clinical study: faecal sRAGE was differentially expressed in patients with active IBD compared with IBD in remission, patients with IBD excluded or healthy controls.ConclusionPre-clinical changes in mouse models can identify early pathways in the development of chronic inflammation that human studies cannot. We identified the decoy receptor sRAGE as a potential mechanism for protection against chronic inflammation in colitis in mice and humans. We propose that the RAGE pathway is clinically relevant in the onset of chronic colitis, and that further study of sRAGE in IBD may provide a novel diagnostic and therapeutic target.
Published: 2019
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42. Using precision microbiome profiling to develop a biomarker for immune checkpoint inhibitor response and a novel therapeutic

Author: Trevor D. Lawley, David Bruce, Amy Popple, Philippa Corrie, Catherine Booth, Sarah J. Welsh, Mat Robinson, Roy Rabbie, Simon R. Harris, Kevin Vervier, Robyne Hudson, David J. Adams, and Doreen Milne
Subjects: Cancer Research, Biomarker, Oncology, business.industry, Immune checkpoint inhibitors, Cancer research, Medicine, Cancer, Microbiome, business, medicine.disease, Gut microbiome
Abstract: e21546 Background: Four independent groups have demonstrated that the pre-treatment gut microbiome of cancer patients impacts the subsequent response to Immune Checkpoint Inhibitor (ICIs) therapy [1-4]. However, the patient’s outcome was linked to different bacteria in each study, which has limited the development of drug response biomarkers and clinic-first design of novel microbiome-based therapeutics. Methods: The Cambridge (UK) MELRESIST study includes a cohort of advanced melanoma patients receiving approved ICIs. Pretreatment stool samples from MELRESIST were analysed by Microbiotica using shotgun metagenomic sequencing. Microbiotica’s platform comprises the world’s leading Reference Genome Database to give the most comprehensive and precise mapping of the gut microbiome. Results: MELRESIST samples showed an overall difference in the microbiome composition between advanced melanoma patients and healthy donors, but not between patients who did or did not respond to ICIs. However, we did identify a discrete microbiome signature that differentiated responders and non-responders with an accuracy of 93%. We extended this signature by reanalysing three published melanoma cohorts [1-3] using the Microbiotica platform, and a propriety bioinformatic model. The resultant bacterial signature was very accurate at predicting response in all 4 published studies combined (91%), and each cohort individually (82-100%). We validated the model using independent validation cohorts and the signature using lung and renal cancer studies [4]. At the core of our microbiome signature was 9 bacteria most significantly associated with ICI efficacy. All 9 were overrepresented in patients who responded to immunotherapy suggesting high abundance of these bacteria is a central driver of ICI response. A consortium comprised of all 9 strains had very potent anti-tumor efficacy in a cancer syngeneic mouse model. The bacteria also demonstrate multiple interactions with primary human immune cells in vitro leading to dendritic cells activation, Cytotoxic T lymphocyte activation and tumor cell killing. These validate the potential of this consortium as a novel therapy for use in combination with ICIs. Conclusions: We have identified a unique microbiome signature predictive of ICI response in 4 independent melanoma cancer cohorts. This removes a major challenge to the field, and could represent a new highly accurate biomarker with clinical application. Nine core bacteria appear to be driving response, and demonstrate anti-tumor activity in vivo and in vitro. This consortium holds great potential as a co-therapy with ICIs. References:1 Matson V et al, Science (2018) 359:104; 2 Gopalakrishnan V et al, Science (2018) 359:97; 3 Frankel AE et al, Neoplasia (2017) 19:848; 4 Routy B et al, Science (2018) 359:91.
Published: 2021
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43. Characterization and Etiology of Swollen Muzzles in Irradiated Mice

Author: Christie M. Orschell, P. Artur Plett, Jeffrey Parker, Richard A. Venezia, Joseph R. Dynlacht, Debra L. Hickman, Catherine Booth, Joy Garrett, Carol H. Sampson, Hui Lin Chua, Robin Crisler, and Thomas J. MacVittie
Subjects: 0301 basic medicine, Male, Pathology, medicine.medical_specialty, Bone marrow transplantation, Biophysics, macromolecular substances, 03 medical and health sciences, Mice, 0302 clinical medicine, Medicine, Animals, Edema, Radiology, Nuclear Medicine and imaging, Retrospective Studies, Radiation, business.industry, Extramural, digestive system diseases, Mice, Inbred C57BL, Radiation Injuries, Experimental, 030104 developmental biology, Acute Radiation Syndrome, 030220 oncology & carcinogenesis, Face, Etiology, Female, business, Head, Neck, Whole-Body Irradiation
Abstract: Several investigators performing bone marrow transplantation studies have previously reported sporadic increases in mortality that were associated with pronounced swelling in the face, head and neck of mice. Over the past few years, we and others have noted an increasing number of experiments in which mice that have received total-body irradiation (TBI) or partial-body irradiation (PBI) develop swollen muzzles, drastic thickening of the upper lip and redness, bruising and/or swelling around the nose and muzzle and sometimes over the top of the head. We refer to this rapid and extreme swelling after irradiation as swollen muzzle syndrome (SMS). The development of SMS postirradiation is associated with morbidity that occurs earlier than would be expected from the traditional hematopoietic acute radiation syndrome (H-ARS), and has impeded studies in several laboratories attempting to evaluate medical countermeasures (MCM) against radiation. However, little has been done to characterize this somewhat unpredictable radiation effect. To investigate the cause and etiology of SMS, data from three different laboratories collected over a seven-year period from 100 MCM 30-day survival studies using mice from different vendors were retrospectively analyzed to determine the time of onset, progression and incidence of SMS in male and female mice exposed to various doses of ionizing radiation. An additional study compared incidence and etiology of SMS in mice from two different vendors (identified as vendors A and B) after exposure to the LD
Published: 2018

44. The Delayed Effects of Acute Radiation Syndrome

Author: Catherine Booth, Gregory Tudor, Thomas J. MacVittie, and Barry P. Katz
Subjects: Male, Pathology, medicine.medical_specialty, Epidemiology, Health, Toxicology and Mutagenesis, Crypt, Biology, Radiation Dosage, Sensitivity and Specificity, Gastrointestinal epithelium, Article, Colony-Forming Units Assay, Lethal Dose 50, Mice, Intestine, Small, medicine, Animals, Radiology, Nuclear Medicine and imaging, Longitudinal Studies, Radiometry, Clonogenic assay, Dose-Response Relationship, Drug, Reproducibility of Results, Dose-Response Relationship, Radiation, Small intestine, Transplantation, Disease Models, Animal, Intestinal Diseases, Haematopoiesis, medicine.anatomical_structure, Acute Radiation Syndrome, Organ Specificity, Cancer research, Bone marrow, Stem cell, Whole-Body Irradiation
Abstract: Long term or residual damage post-irradiation has been described for many tissues. In hematopoietic stem cells (HSC) this is only revealed when the HSC are stressed and required to regenerate and repopulate a myeloablated host. Such an assay cannot be used to assess the recovery potential of previously irradiated intestinal stem cells (ISC) due to their incompatibility with transplantation. The best approximation to the HSC assay is the crypt microcolony assay, also based on clonogen survival. In the current study the regenerative capacity of intestinal clonogenic cells in mice that had survived 13 Gy irradiation (with 5% bone marrow shielding to allow survival through the hematopoietic syndrome) and were then aged for 200 days, was compared to previously unirradiated age matched controls. Interestingly, at 200 days following 13 Gy there remained a statistically significant reduction in crypts present in the various small intestinal regions (illustrating that the gastrointestinal epithelium had not fully recovered despite the 200 day interval). However, upon re-irradiation on day 196, those mice previously irradiated had improved crypt survival and regeneration compared to the age matched controls. This was evident in all regions of the small intestine following 11–13 Gy re-exposure. Thus, there were either more clonogens per crypt within those previously irradiated and/or those that were present were more radioresistant (possibly because a subpopulation was more quiescent). This is contrary to the popular belief that previously irradiated animals may have an impaired/delayed regenerative response and be more radiosensitive.
Published: 2015
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45. Citrulline as a Biomarker in the Murine Total-Body Irradiation Model

Author: Catherine Booth, Fei Li, Gregory Tudor, Jace W. Jones, Thomas J. MacVittie, Maureen A. Kane, Ann M. Farese, Yan Tong, and Barry P. Katz
Subjects: Male, Pathology, medicine.medical_specialty, Epidemiology, Health, Toxicology and Mutagenesis, Context (language use), Biology, Radiation Dosage, Sensitivity and Specificity, Article, Mice, chemistry.chemical_compound, Intestinal mucosa, Intestine, Small, medicine, Citrulline, Animals, Radiology, Nuclear Medicine and imaging, Intestinal Mucosa, Radiation Injuries, Gastrointestinal tract, Reproducibility of Results, Acute Radiation Syndrome, Dose-Response Relationship, Radiation, Total body irradiation, Small intestine, Mice, Inbred C57BL, Survival Rate, Disease Models, Animal, Intestinal Diseases, Dose–response relationship, medicine.anatomical_structure, chemistry, Biomarkers, Whole-Body Irradiation
Abstract: The use of plasma citrulline as a biomarker for gastrointestinal acute radiation syndrome via exposure to total-body irradiation in a murine model was investigated. The radiation exposure covered lethal, mid-lethal, and sub-lethal gastrointestinal acute radiation syndrome. Plasma citrulline profiles were generated over the first 6 days following total-body irradiation exposure of 6–15 Gy. In addition, plasma citrulline was comprehensively evaluated in the context of matching small intestine citrulline and histopathology. Higher plasma citrulline was significantly associated with lower irradiation doses over the first 6 days following the irradiation insult. Furthermore, higher plasma citrulline was significantly associated with higher crypt survival. The correlation of the plasma citrulline to crypt survival was more robust for higher irradiation doses and for later time points. The data suggested plasma citrulline was most informative for reflecting gastrointestinal injury resulting from exposure to 9–15 Gy total-body irradiation covering time-points 2 to 5 days post the irradiation insult.
Published: 2015
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46. Citrulline as a Biomarker in the Non-human Primate Total- and Partial-body Irradiation Models

Author: Catherine Booth, Gregory Tudor, Maureen A. Kane, Alexander Bennett, Ann M. Farese, Kim G. Hankey, Claire L. Carter, Thomas J. MacVittie, and Jace W. Jones
Subjects: Male, medicine.medical_specialty, Pathology, Gastrointestinal Diseases, Epidemiology, Enterocyte, Health, Toxicology and Mutagenesis, Gastrointestinal Injury, Radiation Dosage, Sensitivity and Specificity, Gastroenterology, Article, chemistry.chemical_compound, Internal medicine, medicine, Citrulline, Animals, Radiology, Nuclear Medicine and imaging, Irradiation, Radiation Injuries, Survival rate, business.industry, Reproducibility of Results, Macaca mulatta, Survival Rate, Disease Models, Animal, medicine.anatomical_structure, chemistry, Biomarker (medicine), Histopathology, Bone marrow, business, Biomarkers, Whole-Body Irradiation
Abstract: The use of plasma citrulline as a biomarker for acute and prolonged gastrointestinal injury via exposure to total- and partial-body irradiation (6 MV LINAC-derived photons; 0.80 Gy min−1) in nonhuman primate models was investigated. The irradiation exposure covered gastrointestinal injuries spanning lethal, mid-lethal, and sub-lethal doses. The acute gastrointestinal injury was assessed via measurement of plasma citrulline and small intestinal histopathology over the first 15 days following radiation exposure and included total-body irradiation at 13.0 Gy, 10.5 Gy, and 7.5 Gy and partial-body irradiation at 11.0 Gy with 5% bone marrow sparing. The dosing schemes of 7.5 Gy total-body irradiation and 11.0 Gy partial-body irradiation included time points out to day 60 and day 180, respectively, which allowed for correlation of plasma citrulline to prolonged gastrointestinal injury and survival. Plasma citrulline values were radiation-dependent for all radiation doses under consideration with nadir values ranging from 63–80 % lower than radiation-naïve NHP plasma. The nadir values were observed at day 5 to 7 post irradiation. Longitudinal plasma citrulline profiles demonstrated prolonged gastrointestinal injury resulting from acute high-dose irradiation had long lasting effects on enterocyte function. Moreover, plasma citrulline did not discriminate between total-body or partial-body irradiation over the first 15 days following irradiation and was not predictive of survival based on the radiation models considered herein.
Published: 2015
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47. Results of a multicentre UK-wide retrospective study evaluating the efficacy of brentuximab vedotin in relapsed, refractory classical Hodgkin lymphoma in the transplant naive setting

Author: Catherine Booth, Karl S. Peggs, Fiona Miall, David W Eyre, Graham P. Collins, Wendy Osborne, Adam Gibb, Elizabeth H Phillips, Gillian Stewart, John Radford, Kirit M. Ardeshna, Suzanne Allibone, Kim Linton, Cathy Burton, Arvind Arumainathan, Rifca Ledieu, Shireen Kassam, and Toby A. Eyre
Subjects: Oncology, Male, Immunoconjugates, medicine.medical_treatment, Salvage therapy, 0302 clinical medicine, Nodular sclerosis, immune system diseases, Recurrence, hemic and lymphatic diseases, Antineoplastic Combined Chemotherapy Protocols, Brentuximab vedotin, Brentuximab Vedotin, Manchester Cancer Research Centre, Hematology, Middle Aged, Hodgkin Disease, Multicenter Study, surgical procedures, operative, Treatment Outcome, 030220 oncology & carcinogenesis, Disease Progression, Female, medicine.drug, Adult, medicine.medical_specialty, Adolescent, Antineoplastic Agents, 03 medical and health sciences, Young Adult, Refractory, Internal medicine, medicine, Journal Article, Humans, Survival analysis, Aged, Retrospective Studies, Salvage Therapy, Chemotherapy, business.industry, Contraindications, ResearchInstitutes_Networks_Beacons/mcrc, Retrospective cohort study, medicine.disease, Survival Analysis, Transplantation, business, human activities, 030215 immunology, Stem Cell Transplantation
Abstract: Relapsed or refractory classical Hodgkin lymphoma (cHL) is associated with a poor outcome when standard chemotherapy fails. Brentuximab vedotin (BV) is an anti-CD30 monoclonal antibody-drug conjugate licensed for use at relapse after autologous stem cell transplant (ASCT) or following two prior therapies in those unsuitable for ASCT. There are limited data assessing the ability of BV to enable curative SCT. We performed a UK-wide retrospective study of 99 SCT-naïve relapsed/refractory cHL. All had received 2 prior lines and were deemed fit for transplant but had an insufficient remission to proceed. The median age was 32 years. Most had nodular sclerosis subtype, Eastern Cooperative Oncology Group performance status 0-1 and advanced stage disease. The median progression-free survival (PFS) was 5·6 months and median overall survival (OS) was 37·2 months. The overall response rate was 56% (29% complete response; 27% partial response). 61% reached SCT: 34% immediately post-BV and 27% following an inadequate BV response but were salvaged and underwent deferred SCT. Patients consolidated with SCT had a superior PFS and OS to those not receiving SCT (P < 0·001). BV is an effective, non-toxic bridge to immediate SCT in 34% and deferred SCT in 27%. 39% never reached SCT with a PFS of 3·0 months, demonstrating the unmet need to improve outcomes in those unsuitable for SCT post-BV.
Published: 2017
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48. Identification and Quantitation of Biomarkers for Radiation-induced Injury via Mass Spectrometry

Author: Thomas J. MacVittie, Gregory Tudor, Robert K. Ernst, Jace W. Jones, Zeljko Vujaskovic, Alison J. Scott, Isabel L. Jackson, Maureen A. Kane, P. Xu, and Catherine Booth
Subjects: Male, Biomarker identification, Pathology, medicine.medical_specialty, Epidemiology, Health, Toxicology and Mutagenesis, Tretinoin, Radiation induced, Computational biology, Biology, Mass spectrometry, Mass Spectrometry, Article, Mice, Metabolomics, medicine, Animals, Radiology, Nuclear Medicine and imaging, Biomarker discovery, Lung, Acute Radiation Syndrome, Mice, Inbred C57BL, Radiation Injuries, Experimental, Targeted mass spectrometry, Citrulline, Identification (biology), Biomarkers
Abstract: Biomarker identification and validation for radiation exposure is a rapidly expanding field encompassing the need for well defined animal models and advanced analytical techniques. The resources within the consortium, Medical Countermeasures Against Radiological Threats (MCART), provide a unique opportunity for accessing well defined animal models that simulate the key sequelae of the acute radiation syndrome and the delayed effects of acute radiation exposure. Likewise, the use of mass spectrometry-based analytical techniques for biomarker discovery and validation enables a robust analytical platform that is amenable to a variety of sample matrices and considered the benchmark for biomolecular identification and quantitation. Herein, the authors demonstrate the use of two targeted mass spectrometry approaches to link established MCART animal models to identified metabolite biomarkers. Circulating citrulline concentration was correlated to gross histological gastrointestinal tissue damage, and retinoic acid production in lung tissue was established to be reduced at early and late time points post high dose irradiation. Going forward, the use of mass spectrometry-based metabolomics coupled to well defined animal models provides the unique opportunity for comprehensive biomarker discovery.
Published: 2014
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49. Analytical approaches for quantification of a Nrf2 pathway activator: overcoming bioanalytical challenges to support a toxicity study

Author: James F. Callahan, Hongxing Yan, Roderick S. Davis, Venkatraman Junnotula, Catherine Booth-Genthe, Hermes Licea Perez, Yolanda Sanchez, Hong Nie, Dana Knecht, and Jeffrey C. Boehm
Subjects: Male, Edman degradation, Dimethyl fumarate, NF-E2-Related Factor 2, Chemistry, Activator (genetics), Glutathione, Reference Standards, Biochemistry, Rats, Analytical Chemistry, chemistry.chemical_compound, Tandem Mass Spectrometry, Area Under Curve, Toxicity Tests, Electrochemistry, Animals, Environmental Chemistry, Moiety, Spectroscopy, Chromatography, Liquid, Macromolecule, Cysteine, Sulforaphane
Abstract: Activation of the Nrf2 stress pathway is known to play an important role in the defense mechanism against electrophilic and oxidative damage to biological macromolecules (DNA, lipids, and proteins). Chemical inducers of Nrf2 such as sulforaphane, dimethyl fumarate (Tecfidera®), CDDO-Me (bardoxolone-methyl), and 3-(dimethylamino)-4-((3-isothiocyanatopropyl)(methyl)amino)cyclobut-3-ene-1,2-dione (a synthetic sulforaphane analogue; will be referred to as 1) have the ability to react with Keap1 cysteine residues, leading to activation of the Antioxidant Response Element (ARE). Due to their electrophilic nature and poor matrix stability, these compounds represent great challenges when developing bioanalytical methods to evaluate in vivo exposure. 1 like SFN reacts rapidly with glutathione (GSH) and nucleophilic groups in proteins to form covalent adducts. In this work, three procedures were developed to estimate the exposure of 1 in a non-GLP 7 day safety study in rats: (1) protein precipitation of blood samples with methanol containing the free thiol trapping reagent 4-fluoro-7-aminosulfonylbenzofurazan (ABD-F) to measure GSH- and N-acetylcysteine conjugated metabolites of 1; (2) an Edman degradation procedure to cleave and analyze N-terminal adducts of 1 at the valine moiety; and (3) treatment with ammonium hydroxide to measure circulating free- and all sulfhydryl bound 1.
Published: 2014
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50. Abstract 1061: Animal models of prostate cancer

Author: Frida Ponthan, Matthew Brown, Emma Playle, and Catherine Booth
Subjects: Cancer Research, Oncology
Abstract: Background: Prostate cancer is the most common male malignancy and the second most common cause of cancer-related deaths in the United Kingdom and the United States. It’s a heterogeneous disease caused by different genetic mutations where the majority of prostate cancers are indolent and slow growing. However, some tumours are aggressive with the ability to invade local structures and metastasise leading to poor clinical outcome. One of the most important aspects in anti-cancer drug development is the availability of robust preclinical models which allow the efficacy of novel therapies to be examined. We have developed a reproducible, well growing and robust subcutaneous model of LNCaP with an effective response to both conventional standard of care (SOC) chemotherapy and androgen receptor inhibitors. Materials and Methods: Cells were subcutaneously implanted on the flank of NSG mice and treated with an anti-androgen with or without conventional chemotherapy starting after 15-20 days when the mean tumour volume was 150-200mm3. Animal weight and tumour volume was measured 3x weekly. Androgen receptor (AR) expression was analysed with immunohistochemistry and gene expression analysis before and after treatment. Results: The LNCaP cell line grew well in vivo with 100% take rate. Vehicle treated tumours doubled in size over the first 6 days of treatment and were 4.5x the mean starting volume by day 17 post-treatment start, when the group was removed from the study. Significant tumour growth inhibition was observed in response to both conventional and anti-androgen receptor treatment consistent with the maintained AR expression confirmed by gene expression analysis. Conclusion: This high take rate responsive model represents a significant improvement in subcutaneous xenograft models of prostate cancer, and allows the comparison of AR responsive therapies, alone or in combination with conventional SOC or irradiation. Current protocols employing luciferase transfected cells has also enabled evaluation of metastatic disease. Citation Format: Frida Ponthan, Matthew Brown, Emma Playle, Catherine Booth. Animal models of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1061.
Published: 2019
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