Author: "James, Hadfield" - Searchworks@Jio Institute Digital Library Search Results

114 results on '"James, Hadfield"'

1. Tracing household transmission of SARS-CoV-2 in New Zealand using genomics

Author: Lauren Jelley, Nayyereh Aminisani, Meaghan O’Neill, Tineke Jennings, Jordan Douglas, Srushti Utekar, Helen Johnston, David Welch, James Hadfield, SHIVERS Investigation Team, Joep de Ligt, David Winter, Nigel French, Paul G. Thomas, Richard J. Webby, Sue Huang, and Jemma L. Geoghegan
Subjects: Infectious and parasitic diseases, RC109-216, Biology (General), QH301-705.5
Abstract: Abstract By early 2022, the highly transmissible Omicron variant of SARS-CoV-2 had spread across most of the world. For the first time since the pandemic began, New Zealand was experiencing high levels of community transmission of SARS-CoV-2. We enroled a cohort of households to better understand differences in transmission dynamics among subvariants of Omicron. We enroled 71 households, comprising 289 participants, and aimed to use viral genomes to gain a clearer understanding of variant-specific differences in epidemiological parameters affecting transmission dynamics. Approximately 80% of the households enroled experienced transmission of BA.2, while most of the remaining households had infections with BA.1 or BA.5. Using a logistic regression generalised linear mixed model, we found no difference in household secondary infection rate between Omicron subvariants BA.1, BA.2 and BA.5. Of the households recruited, the vast majority (92%) experienced a single chain of transmission with one inferred introduction. Further, we found that in 48% of the households studied, all household participants became infected following an index case. Most household participants tested positive within a week following an introduction, supporting the seven-day isolation requirement for household contacts that was in place in New Zealand at the time. By integrating genomic and epidemiological data, we show that viral transmission dynamics can be investigated with a higher level of granularity than with epidemiological data alone. Overall, households are a high risk setting for viral transmission in New Zealand.
Published: 2024
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2. Tracing the international arrivals of SARS-CoV-2 Omicron variants after Aotearoa New Zealand reopened its border

Author: Jordan Douglas, David Winter, Andrea McNeill, Sam Carr, Michael Bunce, Nigel French, James Hadfield, Joep de Ligt, David Welch, and Jemma L. Geoghegan
Subjects: Science
Abstract: In March 2022, Aotearoa New Zealand re-opened its border allowing quarantine-free travel for many travellers. Here, the authors describe circulating Omicron sub-variants before and after the reopening of the border and show that the rate of viral introductions grew roughly linearly with the increase in daily international travel.
Published: 2022
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3. Genomic epidemiology of Delta SARS-CoV-2 during transition from elimination to suppression in Aotearoa New Zealand

Author: Lauren Jelley, Jordan Douglas, Xiaoyun Ren, David Winter, Andrea McNeill, Sue Huang, Nigel French, David Welch, James Hadfield, Joep de Ligt, and Jemma L. Geoghegan
Subjects: Science
Abstract: Aotearoa New Zealand pursued a COVID-19 elimination strategy until October 2021 when it moved to a suppression strategy. In this genomic surveillance study, the authors describe spread of the virus during the transition between these strategies, with evidence of substantial undetected community transmission.
Published: 2022
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4. Joint visualization of seasonal influenza serology and phylogeny to inform vaccine composition

Author: Jover Lee, James Hadfield, Allison Black, Thomas R. Sibley, Richard A. Neher, Trevor Bedford, and John Huddleston
Subjects: influenza, serology, phylogeny, interactive, visualization, pandemic, Computer applications to medicine. Medical informatics, R858-859.7
Abstract: Seasonal influenza vaccines must be updated regularly to account for mutations that allow influenza viruses to escape our existing immunity. A successful vaccine should represent the genetic diversity of recently circulating viruses and induce antibodies that effectively prevent infection by those recent viruses. Thus, linking the genetic composition of circulating viruses and the serological experimental results measuring antibody efficacy is crucial to the vaccine design decision. Historically, genetic and serological data have been presented separately in the form of static visualizations of phylogenetic trees and tabular serological results to identify vaccine candidates. To simplify this decision-making process, we have created an interactive tool for visualizing serological data that has been integrated into Nextstrain’s real-time phylogenetic visualization framework, Auspice. We show how the combined interactive visualizations may be used by decision makers to explore the relationships between complex data sets for both prospective vaccine virus selection and retrospectively exploring the performance of vaccine viruses.
Published: 2023
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5. Real-Time Genomics for Tracking Severe Acute Respiratory Syndrome Coronavirus 2 Border Incursions after Virus Elimination, New Zealand

Author: Jordan Douglas, Jemma L. Geoghegan, James Hadfield, Remco Bouckaert, Matthew Storey, Xiaoyun Ren, Joep de Ligt, Nigel French, and David Welch
Subjects: 2019 novel coronavirus disease, coronavirus disease, COVID-19, severe acute respiratory syndrome coronavirus 2, SARS-CoV-2, viruses, Medicine, Infectious and parasitic diseases, RC109-216
Abstract: Since severe acute respiratory syndrome coronavirus 2 was first eliminated in New Zealand in May 2020, a total of 13 known coronavirus disease (COVID-19) community outbreaks have occurred, 2 of which led health officials to issue stay-at-home orders. These outbreaks originated at the border via isolating returnees, airline workers, and cargo vessels. Because a public health system was informed by real-time viral genomic sequencing and complete genomes typically were available within 12 hours of community-based positive COVID-19 test results, every outbreak was well-contained. A total of 225 community cases resulted in 3 deaths. Real-time genomics were essential for establishing links between cases when epidemiologic data could not do so and for identifying when concurrent outbreaks had different origins.
Published: 2021
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6. Fragmentation patterns and personalized sequencing of cell‐free DNA in urine and plasma of glioma patients

Author: Florent Mouliere, Christopher G Smith, Katrin Heider, Jing Su, Ymke van der Pol, Mareike Thompson, James Morris, Jonathan C M Wan, Dineika Chandrananda, James Hadfield, Marta Grzelak, Irena Hudecova, Dominique‐Laurent Couturier, Wendy Cooper, Hui Zhao, Davina Gale, Matthew Eldridge, Colin Watts, Kevin Brindle, Nitzan Rosenfeld, and Richard Mair
Subjects: cell‐free DNA, circulating tumor DNA, fragmentomics, gliomas, liquid biopsy, Medicine (General), R5-920, Genetics, QH426-470
Abstract: Abstract Glioma‐derived cell‐free DNA (cfDNA) is challenging to detect using liquid biopsy because quantities in body fluids are low. We determined the glioma‐derived DNA fraction in cerebrospinal fluid (CSF), plasma, and urine samples from patients using sequencing of personalized capture panels guided by analysis of matched tumor biopsies. By sequencing cfDNA across thousands of mutations, identified individually in each patient’s tumor, we detected tumor‐derived DNA in the majority of CSF (7/8), plasma (10/12), and urine samples (10/16), with a median tumor fraction of 6.4 × 10−3, 3.1 × 10−5, and 4.7 × 10−5, respectively. We identified a shift in the size distribution of tumor‐derived cfDNA fragments in these body fluids. We further analyzed cfDNA fragment sizes using whole‐genome sequencing, in urine samples from 35 glioma patients, 27 individuals with non‐malignant brain disorders, and 26 healthy individuals. cfDNA in urine of glioma patients was significantly more fragmented compared to urine from patients with non‐malignant brain disorders (P = 1.7 × 10−2) and healthy individuals (P = 5.2 × 10−9). Machine learning models integrating fragment length could differentiate urine samples from glioma patients (AUC = 0.80–0.91) suggesting possibilities for truly non‐invasive cancer detection.
Published: 2021
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7. Use of Genomics to Track Coronavirus Disease Outbreaks, New Zealand

Author: Jemma L. Geoghegan, Jordan Douglas, Xiaoyun Ren, Matthew Storey, James Hadfield, Olin K. Silander, Nikki E. Freed, Lauren Jelley, Sarah Jefferies, Jillian Sherwood, Shevaun Paine, Sue Huang, Andrew Sporle, Michael G. Baker, David R. Murdoch, Alexei J. Drummond, David Welch, Colin R. Simpson, Nigel French, Edward C. Holmes, and Joep de Ligt
Subjects: 2019 novel coronavirus disease, coronavirus disease, COVID-19, severe acute respiratory syndrome coronavirus 2, SARS-CoV-2, viruses, Medicine, Infectious and parasitic diseases, RC109-216
Abstract: Real-time genomic sequencing has played a major role in tracking the global spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), contributing greatly to disease mitigation strategies. In August 2020, after having eliminated the virus, New Zealand experienced a second outbreak. During that outbreak, New Zealand used genomic sequencing in a primary role, leading to a second elimination of the virus. We generated genomes from 78% of the laboratory-confirmed samples of SARS-CoV-2 from the second outbreak and compared them with the available global genomic data. Genomic sequencing rapidly identified that virus causing the second outbreak in New Zealand belonged to a single cluster, thus resulting from a single introduction. However, successful identification of the origin of this outbreak was impeded by substantial biases and gaps in global sequencing data. Access to a broader and more heterogenous sample of global genomic data would strengthen efforts to locate the source of any new outbreaks.
Published: 2021
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8. Genomic Evidence of In-Flight Transmission of SARS-CoV-2 Despite Predeparture Testing

Author: Tara Swadi, Jemma L. Geoghegan, Tom Devine, Caroline McElnay, Jillian Sherwood, Phil Shoemack, Xiaoyun Ren, Matt Storey, Sarah Jefferies, Erasmus Smit, James Hadfield, Aoife Kenny, Lauren Jelley, Andrew Sporle, Andrea McNeill, G. Edwin Reynolds, Kip Mouldey, Lindsay Lowe, Gerard Sonder, Alexei J. Drummond, Sue Huang, David Welch, Edward C. Holmes, Nigel French, Colin R. Simpson, and Joep de Ligt
Subjects: 2019 novel coronavirus disease, coronavirus disease, COVID-19, severe acute respiratory syndrome coronavirus 2, SARS-CoV-2, viruses, Medicine, Infectious and parasitic diseases, RC109-216
Abstract: Since the first wave of coronavirus disease in March 2020, citizens and permanent residents returning to New Zealand have been required to undergo managed isolation and quarantine (MIQ) for 14 days and mandatory testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As of October 20, 2020, of 62,698 arrivals, testing of persons in MIQ had identified 215 cases of SARS-CoV-2 infection. Among 86 passengers on a flight from Dubai, United Arab Emirates, that arrived in New Zealand on September 29, test results were positive for 7 persons in MIQ. These passengers originated from 5 different countries before a layover in Dubai; 5 had negative predeparture SARS-CoV-2 test results. To assess possible points of infection, we analyzed information about their journeys, disease progression, and virus genomic data. All 7 SARS-CoV-2 genomes were genetically identical, except for a single mutation in 1 sample. Despite predeparture testing, multiple instances of in-flight SARS-CoV-2 transmission are likely.
Published: 2021
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9. Genomic epidemiology reveals transmission patterns and dynamics of SARS-CoV-2 in Aotearoa New Zealand

Author: Jemma L. Geoghegan, Xiaoyun Ren, Matthew Storey, James Hadfield, Lauren Jelley, Sarah Jefferies, Jill Sherwood, Shevaun Paine, Sue Huang, Jordan Douglas, Fábio K. Mendes, Andrew Sporle, Michael G. Baker, David R. Murdoch, Nigel French, Colin R. Simpson, David Welch, Alexei J. Drummond, Edward C. Holmes, Sebastián Duchêne, and Joep de Ligt
Subjects: Science
Abstract: New Zealand implemented stringent COVID-19 control measures early after identification of its first case. Here, the authors perform whole genome sequencing of samples taken until 22 May 2020 and find high viral diversity indicative of multiple separate introductions and limited community transmission.
Published: 2020
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10. High-throughput sequencing of SARS-CoV-2 in wastewater provides insights into circulating variants.

Author: Rafaela S. Fontenele, Simona Kraberger, James Hadfield, Erin M. Driver, Devin Bowes, LaRinda A. Holland, Temitope O. Faleye, Sangeet Adhikari, Rahul Kumar, Rosa Inchausti, Wydale K. Holmes, Stephanie Deitrick, Philip Brown, Darrell Duty, Aruni Bhatnagar, Ray A. Yeager, Rochelle H. Holm, Kevin Dixon, Tim Constantine, Melissa A. Wilson, Efrem S. Lim, Xiaofang Jiang, Rolf U. Halden, Matthew Scotch, and Arvind Varsani
Published: 2021

11. Independence of HIF1a and androgen signaling pathways in prostate cancer

Author: Maxine G. B. Tran, Becky A. S. Bibby, Lingjian Yang, Franklin Lo, Anne Y. Warren, Deepa Shukla, Michelle Osborne, James Hadfield, Thomas Carroll, Rory Stark, Helen Scott, Antonio Ramos-Montoya, Charlie Massie, Patrick Maxwell, Catharine M. L. West, Ian G. Mills, and David E. Neal
Subjects: Prostate cancer, Androgen signaling, HIF1a signaling, Hypoxia, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract: Abstract Background Therapeutic targeting of the androgen signaling pathway is a mainstay treatment for prostate cancer. Although initially effective, resistance to androgen targeted therapies develops followed by disease progression to castrate-resistant prostate cancer (CRPC). Hypoxia and HIF1a have been implicated in the development of resistance to androgen targeted therapies and progression to CRCP. The interplay between the androgen and hypoxia/HIF1a signaling axes was investigated. Methods In vitro stable expression of HIF1a was established in the LNCaP cell line by physiological induction or retroviral transduction. Tumor xenografts with stable expression of HIF1a were established in castrated and non-castrated mouse models. Gene expression analysis identified transcriptional changes in response to androgen treatment, hypoxia and HIF1a. The binding sites of the AR and HIF transcription factors were identified using ChIP-seq. Results Androgen and HIF1a signaling promoted proliferation in vitro and enhanced tumor growth in vivo. The stable expression of HIF1a in vivo restored tumor growth in the absence of endogenous androgens. Hypoxia reduced AR binding sites whereas HIF binding sites were increased with androgen treatment under hypoxia. Gene expression analysis identified seven genes that were upregulated both by AR and HIF1a, of which six were prognostic. Conclusions The oncogenic AR, hypoxia and HIF1a pathways support prostate cancer development through independent signaling pathways and transcriptomic profiles. AR and hypoxia/HIF1a signaling pathways independently promote prostate cancer progression and therapeutic targeting of both pathways simultaneously is warranted.
Published: 2020
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12. Development and validation of blood tumor mutational burden reference standards

Author: Eun-Ang, Raiber-Moreau, Guillem, Portella, Matthew G, Butler, Omoshile, Clement, Yves, Konigshofer, and James, Hadfield
Subjects: Cancer Research, Genetics
Abstract: Tumor mutational burden (TMB), measured by exome or panel sequencing of tumor tissue or blood (bTMB), is a potential predictive biomarker for treatment benefit in patients with various cancer types receiving immunotherapy targeting checkpoint pathways. However, significant variability in TMB measurement has been observed. We developed contrived bTMB reference materials using DNA from tumor cell lines and donor-matched lymphoblastoid cell lines to support calibration and alignment across laboratories and platforms. Contrived bTMB reference materials were developed using genomic DNA from lung tumor cell lines blended into donor-matched lymphoblastoid cell lines at 0.5% and 2% tumor content, fragmented and size-selected to mirror the size profile of circulating cell-free tumor DNA with TMB scores of 7, 9, 20, and 26 mut/Mb. Variant allele frequency (VAF) and bTMB scores were assessed using PredicineATLAS and GuardantOMNI next-generation sequencing assays. DNA fragment sizes in the contrived reference samples were similar to those found within patient plasma-derived cell-free DNA, and mutational patterns aligned with those in the parental tumor lines. For the 7, 20, and 26 mut/Mb contrived reference samples with 2% tumor content, bTMB scores estimated using either assay aligned with expected scores from the parental tumor cell lines and showed good reproducibility. A bioinformatic filtration step was required to account for low-VAF artifact variants. We demonstrate the feasibility and challenges of producing and using bTMB reference standards across a range of bTMB levels, and how such standards could support the calibration and validation of bTMB platforms and help harmonization between panels and laboratories.
Published: 2022

13. Characterising the epidemic spread of influenza A/H3N2 within a city through phylogenetics.

Author: Nicola F Müller, Daniel Wüthrich, Nina Goldman, Nadine Sailer, Claudia Saalfrank, Myrta Brunner, Noémi Augustin, Helena Mb Seth-Smith, Yvonne Hollenstein, Mohammedyaseen Syedbasha, Daniela Lang, Richard A Neher, Olivier Dubuis, Michael Naegele, Andreas Buser, Christian H Nickel, Nicole Ritz, Andreas Zeller, Brian M Lang, James Hadfield, Trevor Bedford, Manuel Battegay, Rita Schneider-Sliwa, Adrian Egli, and Tanja Stadler
Subjects: Immunologic diseases. Allergy, RC581-607, Biology (General), QH301-705.5
Abstract: Infecting large portions of the global population, seasonal influenza is a major burden on societies around the globe. While the global source sink dynamics of the different seasonal influenza viruses have been studied intensively, its local spread remains less clear. In order to improve our understanding of how influenza is transmitted on a city scale, we collected an extremely densely sampled set of influenza sequences alongside patient metadata. To do so, we sequenced influenza viruses isolated from patients of two different hospitals, as well as private practitioners in Basel, Switzerland during the 2016/2017 influenza season. The genetic sequences reveal that repeated introductions into the city drove the influenza season. We then reconstruct how the effective reproduction number changed over the course of the season. While we did not find that transmission dynamics in Basel correlate with humidity or school closures, we did find some evidence that it may positively correlated with temperature. Alongside the genetic sequence data that allows us to see how individual cases are connected, we gathered patient information, such as the age or household status. Zooming into the local transmission outbreaks suggests that the elderly were to a large extent infected within their own transmission network. In the remaining transmission network, our analyses suggest that school-aged children likely play a more central role than pre-school aged children. These patterns will be valuable to plan interventions combating the spread of respiratory diseases within cities given that similar patterns are observed for other influenza seasons and cities.
Published: 2020
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14. The Seattle Flu Study: a multiarm community-based prospective study protocol for assessing influenza prevalence, transmission and genomic epidemiology

Author: Michael Jackson, Kira Newman, Amanda Adler, Helen Y Chu, Michael Boeckh, Janet A Englund, Michael Famulare, Barry Lutz, Deborah A Nickerson, Mark Rieder, Lea M Starita, Elisabeth Brandstetter, Chris D Frazer, Peter D Han, Reena K Gulati, James Hadfield, Anahita Kiavand, Louise E Kimball, Kirsten Lacombe, Thomas R Sibley, Jennifer K Logue, Victoria Rachel Lyon, Caitlin R Wolf, Monica Zigman Suchsland, Jay Shendure, and Trevor Bedford
Subjects: Medicine
Abstract: Introduction Influenza epidemics and pandemics cause significant morbidity and mortality. An effective response to a potential pandemic requires the infrastructure to rapidly detect, characterise, and potentially contain new and emerging influenza strains at both an individual and population level. The objective of this study is to use data gathered simultaneously from community and hospital sites to develop a model of how influenza enters and spreads in a population.Methods and analysis Starting in the 2018–2019 season, we have been enrolling individuals with acute respiratory illness from community sites throughout the Seattle metropolitan area, including clinics, childcare facilities, Seattle-Tacoma International Airport, workplaces, college campuses and homeless shelters. At these sites, we collect clinical data and mid-nasal swabs from individuals with at least two acute respiratory symptoms. Additionally, we collect residual nasal swabs and data from individuals who seek care for respiratory symptoms at four regional hospitals. Samples are tested using a multiplex molecular assay, and influenza whole genome sequencing is performed for samples with influenza detected. Geospatial mapping and computational modelling platforms are in development to characterise the regional spread of influenza and other respiratory pathogens.Ethics and dissemination The study was approved by the University of Washington’s Institutional Review Board (STUDY00006181). Results will be disseminated through talks at conferences, peer-reviewed publications and on the study website (www.seattleflu.org).
Published: 2020
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15. Differentiation dynamics of mammary epithelial cells revealed by single-cell RNA sequencing

Author: Karsten Bach, Sara Pensa, Marta Grzelak, James Hadfield, David J. Adams, John C. Marioni, and Walid T. Khaled
Subjects: Science
Abstract: There is a need to understand how mammary epithelial cells respond to changes at various developmental stages. Here, the authors use single-cell RNA sequencing of mammary epithelial cells at different adult developmental stages, identifying different cell types and charting their developmental trajectory.
Published: 2017
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16. Mycoplasma genitalium: whole genome sequence analysis, recombination and population structure

Author: Maria C. Fookes, James Hadfield, Simon Harris, Surendra Parmar, Magnus Unemo, Jørgen S. Jensen, and Nicholas R. Thomson
Subjects: Mycoplasma genitalium genomics and phylogenetics, STIs, Azithromycin resistance, Mycoplasma genitalium, Biotechnology, TP248.13-248.65, Genetics, QH426-470
Abstract: Abstract Background Although Mycoplasma genitalium is a common sexually transmitted pathogen causing clinically distinct diseases both in male and females, few genomes have been sequenced up to now, due mainly to its fastidious nature and slow growth. Hence, we lack a robust phylogenetic framework to provide insights into the population structure of the species. Currently our understanding of the nature and diversity of M. genitalium relies on molecular tests targeting specific genes or regions of the genome and knowledge is limited by a general under-testing internationally. This is set against a background of drug resistance whereby M. genitalium has developed resistance to mainly all therapeutic antimicrobials. Results We sequenced 28 genomes of Mycoplasma genitalium from temporally (1980–2010) and geographically (Europe, Japan, Australia) diverse sources. All the strain showed essentially the same genomic content without any accessory regions found. However, we identified extensive recombination across their genomes with a total of 25 regions showing heightened levels of SNP density. These regions include the MgPar loci, associated with host interactions, as well as other genes that could also be involved in this role. Using these data, we generated a robust phylogeny which shows that there are two main clades with differing degrees of genomic variability. SNPs found in region V of 23S rRNA and parC were consistent with azithromycin/erythromycin and fluoroquinolone resistances, respectively, and with their phenotypic MIC data. Conclusions The sequence data here generated is essential for designing rational approaches to type and track Mycoplasma genitalium as antibiotic resistance increases. It represents a first approach to its population genetics to better appreciate the role of this organism as a sexually transmitted pathogen.
Published: 2017
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17. The Open-Fracture Patient Evaluation Nationwide (OPEN) study

Author: James Hadfield, Muhamed Farhan-Alanie, Robert John Henry Winstanley, Will Eardley, Keith Allison, Samuel Newman, Setthasorn Zhi Yang Ooi, Abhinav Singh, Jonathan Taylor, Marios Ghobrial, Zain Elahi, Hamez Gacaferi, Kate Atkinson, and Christopher Bretherton
Subjects: Tibial Fractures, Fracture Fixation, Internal, Fractures, Open, Treatment Outcome, Humans, Surgical Wound Infection, Orthopedics and Sports Medicine, Surgery, United Kingdom, Anti-Bacterial Agents, Follow-Up Studies, Retrospective Studies
Abstract: Aims The Open-Fracture Patient Evaluation Nationwide (OPEN) study was performed to provide clarity in open fracture management previously skewed by small, specialist centre studies and large, unfocused registry investigations. We report the current management metrics of open fractures across the UK. Method Patients admitted to hospital with an open fracture (excluding phalanges or isolated hand injuries) between 1 June 2021 and 30 September 2021 were included. Institutional information governance approval was obtained at the lead site and all data entered using Research Electronic Data Capture software. All domains of the British Orthopaedic Association Standard for Open Fracture Management were recorded. Results Across 51 centres, 1,175 patients were analyzed. Antibiotics were given to 754 (69.0%) in the emergency department, 240 (22.0%) pre-hospital, and 99 (9.1%) as inpatients. Wounds were photographed in 848 (72.7%) cases. Median time to first surgery was 16 hrs 14 mins (interquartile range (IQR) 8 hrs 29 mins to 23 hrs 19 mins). Complex injuries were operated on sooner (median 12 hrs 51 mins (IQR 4 hrs 36 mins to 21 hrs 14 mins)). Of initial procedures, 1,053 (90.3%) occurred between 8am and 8pm. A consultant orthopaedic surgeon was present at 1,039 (89.2%) first procedures. In orthoplastic centres, a consultant plastic surgeon was present at 465 (45.1%) first procedures. Overall, 706 (60.8%) patients required a single operation. At primary debridement, 798 (65.0%) fractures were definitively fixed, while 734 (59.8%) fractures had fixation and coverage in one operation through direct closure or soft-tissue coverage. Negative pressure wound therapy was used in 235 (67.7%) staged procedures. Following wound closure or soft-tissue cover, 509 (47.0%) patients received antibiotics for a median of three days (IQR 1 to 7). Conclusion OPEN provides an insight into care across the UK and different levels of hospital for open fractures. Patients are predominantly operated on promptly, in working hours, and at specialist centres. Areas for improvement include combined patient review and follow-up, scheduled operating, earlier definitive soft-tissue cover, and more robust antibiotic husbandry. Cite this article: Bone Joint J 2022;104-B(9):1073–1080.
Published: 2022

18. Twenty years of West Nile virus spread and evolution in the Americas visualized by Nextstrain.

Author: James Hadfield, Anderson F Brito, Daniele M Swetnam, Chantal B F Vogels, Ryan E Tokarz, Kristian G Andersen, Ryan C Smith, Trevor Bedford, and Nathan D Grubaugh
Subjects: Immunologic diseases. Allergy, RC581-607, Biology (General), QH301-705.5
Abstract: It has been 20 years since West Nile virus first emerged in the Americas, and since then, little progress has been made to control outbreaks caused by this virus. After its first detection in New York in 1999, West Nile virus quickly spread across the continent, causing an epidemic of human disease and massive bird die-offs. Now the virus has become endemic to the United States, where an estimated 7 million human infections have occurred, making it the leading mosquito-borne virus infection and the most common cause of viral encephalitis in the country. To bring new attention to one of the most important mosquito-borne viruses in the Americas, we provide an interactive review using Nextstrain: a visualization tool for real-time tracking of pathogen evolution (nextstrain.org/WNV/NA). Nextstrain utilizes a growing database of more than 2,000 West Nile virus genomes and harnesses the power of phylogenetics for students, educators, public health workers, and researchers to visualize key aspects of virus spread and evolution. Using Nextstrain, we use virus genomics to investigate the emergence of West Nile virus in the U S, followed by its rapid spread, evolution in a new environment, establishment of endemic transmission, and subsequent international spread. For each figure, we include a link to Nextstrain to allow the readers to directly interact with and explore the underlying data in new ways. We also provide a brief online narrative that parallels this review to further explain the data and highlight key epidemiological and evolutionary features (nextstrain.org/narratives/twenty-years-of-WNV). Mirroring the dynamic nature of outbreaks, the Nextstrain links provided within this paper are constantly updated as new West Nile virus genomes are shared publicly, helping to stay current with the research. Overall, our review showcases how genomics can track West Nile virus spread and evolution, as well as potentially uncover novel targeted control measures to help alleviate its public health burden.
Published: 2019
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19. Data from A View on Drug Development for Cancer Prevention

Author: Susan Galbraith, Barry Crean, Gyorgy Zorenyi, Caroline Germa, Gaia Schiavon, Amaya Gasco Hernandez, Benjamin G. Challis, Eden Harrison, James Hadfield, Tina Sarbajna, Zoltan Szucs, Marianne Ashford, Farzana Walcott, Alula R. Yohannes, Michele Moschetta, and Andrew R. Reynolds
Abstract: Despite some notable successes, there are still relatively few agents approved for cancer prevention. Here we review progress thus far in the development of medicines for cancer prevention, and we outline some key concepts that could further enable or accelerate drug development for cancer prevention in the future. These are summarized under six key themes: (i) unmet clinical need, (ii) patient identification, (iii) risk stratification, (iv) pharmacological intervention, (v) clinical trials, and (vi) health care policy. These concepts, if successfully realized, may help to increase the number of medicines available for cancer prevention.Significance:The huge potential public health benefits of preventing cancer, combined with recent advances in the availability of novel early detection technologies and new treatment modalities, has caused us to revisit the opportunities and challenges associated with developing medicines to prevent cancer. Here we review progress in the field of developing medicines to prevent cancer to date, and we present a series of ideas that might help in the development of more medicines to prevent cancer in the future.
Published: 2023

20. Supplementary Tables S1-S3 from A View on Drug Development for Cancer Prevention

Author: Susan Galbraith, Barry Crean, Gyorgy Zorenyi, Caroline Germa, Gaia Schiavon, Amaya Gasco Hernandez, Benjamin G. Challis, Eden Harrison, James Hadfield, Tina Sarbajna, Zoltan Szucs, Marianne Ashford, Farzana Walcott, Alula R. Yohannes, Michele Moschetta, and Andrew R. Reynolds
Abstract: Tables that provide additional information to support the main article
Published: 2023

21. Chlamydia trachomatis from Australian Aboriginal people with trachoma are polyphyletic composed of multiple distinctive lineages

Author: Patiyan Andersson, Simon R. Harris, Helena M. B. Seth Smith, James Hadfield, Colette O’Neill, Lesley T. Cutcliffe, Fiona P. Douglas, L. Valerie Asche, John D. Mathews, Susan I. Hutton, Derek S. Sarovich, Steven Y. C. Tong, Ian N. Clarke, Nicholas R. Thomson, and Philip M. Giffard
Subjects: Science
Abstract: Chlamydia trachomatis isolates causing a blinding disease (trachoma) form a single lineage that is different from the lineages causing urogenital infections. Here, Andersson et al. show however that trachoma isolates from Australia are more closely related to urogenital strains than to other trachoma isolates.
Published: 2016
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22. The Nature and Extent of Plasmid Variation in Chlamydia trachomatis

Author: Charlotte A. Jones, James Hadfield, Nicholas R. Thomson, David W. Cleary, Peter Marsh, Ian N. Clarke, and Colette E. O’Neill
Subjects: chlamydia trachomatis, plasmid, sequencing, genetic variation, evolution, diagnostics, Biology (General), QH301-705.5
Abstract: Chlamydia trachomatis is an obligate intracellular pathogen of humans, causing both the sexually transmitted infection, chlamydia, and the most common cause of infectious blindness, trachoma. The majority of sequenced C. trachomatis clinical isolates carry a 7.5-Kb plasmid, and it is becoming increasingly evident that this is a key determinant of pathogenicity. The discovery of the Swedish New Variant and the more recent Finnish variant highlight the importance of understanding the natural extent of variation in the plasmid. In this study we analysed 524 plasmid sequences from publicly available whole-genome sequence data. Single nucleotide polymorphisms (SNP) in each of the eight coding sequences (CDS) were identified and analysed. There were 224 base positions out of a total 7550 bp that carried a SNP, which equates to a SNP rate of 2.97%, nearly three times what was previously calculated. After normalising for CDS size, CDS8 had the highest SNP rate at 3.97% (i.e., number of SNPs per total number of nucleotides), whilst CDS6 had the lowest at 1.94%. CDS5 had the highest total number of SNPs across the 524 sequences analysed (2267 SNPs), whereas CDS6 had the least SNPs with only 85 SNPs. Calculation of the genetic distances identified CDS6 as the least variable gene at the nucleotide level (d = 0.001), and CDS5 as the most variable (d = 0.007); however, at the amino acid level CDS2 was the least variable (d = 0.001), whilst CDS5 remained the most variable (d = 0.013). This study describes the largest in-depth analysis of the C. trachomatis plasmid to date, through the analysis of plasmid sequence data mined from whole genome sequences spanning 50 years and from a worldwide distribution, providing insights into the nature and extent of existing variation within the plasmid as well as guidance for the design of future diagnostic assays. This is crucial at a time when single-target diagnostic assays are failing to detect natural mutants, putting those infected at risk of a serious long-term and life-changing illness.
Published: 2020
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23. Decision letter: Taxonium, a web-based tool for exploring large phylogenetic trees

Author: Richard A Neher, Art FY Poon, and James Hadfield
Published: 2022

24. The Open-Fracture Patient Evaluation Nationwide (OPEN) study : epidemiology of open fracture care in the UK

Author: James Hadfield, Muhamed Farhan-Alanie, Will Eardley, Charlotte Brookes, Andrew Gray, Samuel Newman, Setthasorn Zhi Yang Ooi, Jonathan Taylor, Reece Walker, Marios Ghobrial, Zain Elahi, Tomisin Omogbehin, Hamez Gacaferi, Kate Atkinson, and Christopher Bretherton
Subjects: General Engineering
Abstract: Aims Understanding of open fracture management is skewed due to reliance on small-number lower limb, specialist unit reports and large, unfocused registry data collections. To address this, we carried out the Open Fracture Patient Evaluation Nationwide (OPEN) study, and report the demographic details and the initial steps of care for patients admitted with open fractures in the UK. Methods Any patient admitted to hospital with an open fracture between 1 June 2021 and 30 September 2021 was included, excluding phalanges and isolated hand injuries. Institutional information governance approval was obtained at the lead site and all data entered using Research Electronic Data Capture. Demographic details, injury, fracture classification, and patient dispersal were detailed. Results In total, 1,175 patients (median age 47 years (interquartile range (IQR) 29 to 65), 61.0% male (n = 717)) were admitted across 51 sites. A total of 546 patients (47.1%) were employed, 5.4% (n = 63) were diabetic, and 28.8% (n = 335) were smokers. In total, 29.0% of patients (n = 341) had more than one injury and 4.8% (n = 56) had two or more open fractures, while 51.3% of fractures (n = 637) occurred in the lower leg. Fractures sustained in vehicle incidents and collisions are common (38.8%; n = 455) and typically seen in younger patients. A simple fall (35.0%; n = 410) is common in older people. Overall, 69.8% (n = 786) of patients were admitted directly to an orthoplastic centre, 23.0% (n = 259) were transferred to an orthoplastic centre after initial management elsewhere, and 7.2% were managed outwith specialist units (n = 81). Conclusion This study describes the epidemiology of open fractures in the UK. For a decade, orthopaedic surgeons have been practicing in a guideline-driven, network system without understanding the patient features, injury characteristics, or dispersal processes of the wider population. This work will inform care pathways as the UK looks to the future of trauma networks and guidelines, and how to optimize care for patients with open fractures. Cite this article: Bone Jt Open 2022;3(10):746–752.
Published: 2022

25. Bioanalyzer chips can be used interchangeably for many analyses of DNA or RNA

Author: Jessica Davies, Tom Denyer, and James Hadfield
Subjects: RNA, NGS, Bioanalyzer, QC, quality control, Biology (General), QH301-705.5
Abstract: The Agilent 2100 Bioanalyzer enables small-scale gel electrophoretic separation of nucleic acids on a microfluidic chip; however, a shortage of chips and an excess of reagents are common issues. Here we explore the compatibility of two commonly used Bioanalyzer reagents with three Bioanalyzer chip types. Microfluidic electrophoretic separation of DNA and RNA using DNA High Sensitivity and RNA 6000 Nano reagents, respectively, was successfully performed on multiple chip types following the assay-specific protocols. For RNA quality and next-generation sequencing (NGS) library length estimation, the Bioanalyzer chips we tested can be used interchangeably. These findings will be valuable for any laboratory using the Agilent Bioanalyzer in a shared facility.
Published: 2016
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26. Ebola Virus Transmission Initiated by Relapse of Systemic Ebola Virus Disease

Author: Karthik Gangavarapu, Amuri Aziza, Ibrahima Socé Fall, Marceline Akonga, Anne W. Rimion, Yannick Tutu Tshia N’kasar, Moussa Moïse Diagne, Fabrice Mambu, Jean Jacques Muyembe Tamfum, Ali Torkamani, Meris Matondo, Daniel Mukadi, Nancy J. Sullivan, Nella Bisento, Antoine Nkuba-Ndaye, Abdoulaye Yam, Emilia Sana-Paka, Elias Salfati, Kristian G. Andersen, Placide Mbala-Kingebeni, Bibiche Nsunda, Bailey White, Martin Faye, Francois Edidi, John Misasi, Marc A. Suchard, Michael R. Wiley, Boubacar Diallo, Catherine Pratt, Faustin Bile, Aurélie Ploquin, Olivier Tshiani, Matthias Pauthner, Steve Ahuka-Mundeke, Sabue Mulangu, Allison Black, Eddy Kinganda-Lusamaki, Amadou A. Sall, Ousmane Faye, Mbusa Mutafali-Ruffin, Lisa E. Hensley, Trevor Bedford, Marie Roseline Darnycka Belizaire, Victor Epaso, Ian Crozier, Donatien Kazadi, James Hadfield, Andrew Rambaut, Mory Keita, and Junior Bula Bula
Subjects: Male, Fatal outcome, viruses, Disease, 030204 cardiovascular system & hematology, medicine.disease_cause, Medical and Health Sciences, Fatal Outcome, 0302 clinical medicine, Recurrence, Viral, 030212 general & internal medicine, Phylogeny, Genome, biology, Transmission (medicine), General Medicine, Ebolavirus, Infectious Diseases, Vesicular stomatitis virus, Ebola, Democratic Republic of the Congo, Infection, Biotechnology, Adult, Viremia, Vaccine Related, 03 medical and health sciences, Rare Diseases, Biodefense, General & Internal Medicine, medicine, Humans, Ebola Vaccines, Ebola virus, Extramural, business.industry, Prevention, Outbreak, Bayes Theorem, biology.organism_classification, medicine.disease, Virology, Emerging Infectious Diseases, Good Health and Well Being, Mutation, Hemorrhagic Fever, RNA, Immunization, business
Abstract: During the 2018-2020 Ebola virus disease (EVD) outbreak in North Kivu province in the Democratic Republic of Congo, EVD was diagnosed in a patient who had received the recombinant vesicular stomatitis virus-based vaccine expressing a ZEBOV glycoprotein (rVSV-ZEBOV) (Merck). His treatment included an Ebola virus (EBOV)-specific monoclonal antibody (mAb114), and he recovered within 14 days. However, 6 months later, he presented again with severe EVD-like illness and EBOV viremia, and he died. We initiated epidemiologic and genomic investigations that showed that the patient had had a relapse of acute EVD that led to a transmission chain resulting in 91 cases across six health zones over 4 months. (Funded by the Bill and Melinda Gates Foundation and others.).
Published: 2021

27. Integration of genomic sequencing into the response to the Ebola virus outbreak in Nord Kivu, Democratic Republic of the Congo

Author: Francois Edidi-Atani, Moussa Moïse Diagne, Meris Matondo-Kuamfumu, Junior Bulabula, Kristian G. Andersen, Bibiche Nsunda, Eric Delaporte, Bailey White, Amadou A. Sall, Trevor Bedford, Matthias Pauthner, Martine Peeters, Nella Bisento, Jean Jacques Muyembe Tamfum, Marceline Akonga, James Hadfield, Daniel Mukadi, Michael R. Wiley, Gustavo Palacios, Fabrice Mambu-Mbika, Amuri Aziza, Nicholas Di Paola, Steve Ahuka-Mundeke, Eddy Kinganda-Lusamaki, Catherine Pratt, Ousmane Faye, Martin Faye, Allison Black, Placide Mbala-Kingebeni, University of Kinshasa (UNIKIN), Institut National de Recherche Biomédicale [Kinshasa] (INRB), University of Washington [Seattle], Fred Hutchinson Cancer Research Center [Seattle] (FHCRC), University of Nebraska Medical Center, University of Nebraska System, Institut Pasteur de Dakar, Réseau International des Instituts Pasteur (RIIP), Université Cheikh Anta Diop [Dakar, Sénégal] (UCAD), The Scripps Research Institute [La Jolla, San Diego], Recherches Translationnelles sur le VIH et les maladies infectieuses endémiques er émergentes (TransVIHMI), Institut de Recherche pour le Développement (IRD)-Université de Yaoundé I-Université Cheikh Anta Diop [Dakar, Sénégal] (UCAD)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), and Retiveau, Nolwenn
Subjects: 0301 basic medicine, MESH: Sequence Analysis, DNA, medicine.medical_specialty, Genomic data, MESH: Reinfection, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Genome, Viral, Disease, medicine.disease_cause, Genome, Article, General Biochemistry, Genetics and Molecular Biology, Disease Outbreaks, 03 medical and health sciences, MESH: Spatio-Temporal Analysis, Spatio-Temporal Analysis, 0302 clinical medicine, Recurrence, Environmental health, MESH: Congo, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, medicine, MESH: Ebola Vaccines, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Disease Outbreaks, MESH: Ebolavirus, Ebola Vaccines, MESH: Phylogeny, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Phylogeny, Ebola virus, Transmission (medicine), MESH: Genomics, Genomic sequencing, Public health, Outbreak, Genomics, Sequence Analysis, DNA, General Medicine, Hemorrhagic Fever, Ebola, Ebolavirus, MESH: Recurrence, 030104 developmental biology, Geography, Congo, Reinfection, 030220 oncology & carcinogenesis, MESH: Hemorrhagic Fever, Ebola, MESH: Genome, Viral
Abstract: On 1 August 2018, the Democratic Republic of the Congo (DRC) declared its tenth Ebola virus disease (EVD) outbreak. To aid the epidemiologic response, the Institut National de Recherche Biomedicale (INRB) implemented an end-to-end genomic surveillance system, including sequencing, bioinformatic analysis and dissemination of genomic epidemiologic results to frontline public health workers. We report 744 new genomes sampled between 27 July 2018 and 27 April 2020 generated by this surveillance effort. Together with previously available sequence data (n = 48 genomes), these data represent almost 24% of all laboratory-confirmed Ebola virus (EBOV) infections in DRC in the period analyzed. We inferred spatiotemporal transmission dynamics from the genomic data as new sequences were generated, and disseminated the results to support epidemiologic response efforts. Here we provide an overview of how this genomic surveillance system functioned, present a full phylodynamic analysis of 792 Ebola genomes from the Nord Kivu outbreak and discuss how the genomic surveillance data informed response efforts and public health decision making. Phylogeographic analysis of 792 Ebola virus genomes from the 2018 oubreak in the Democratic Republic of the Congo integrated into an end-to-end surveillance program demonstrates the feasibility of using genomic sequencing data to inform the public health epidemic response in near-real time.
Published: 2021

28. Genomic Evidence of In-Flight Transmission of SARS-CoV-2 Despite Predeparture Testing

Author: Aoife Kenny, Nigel P. French, Alexei J. Drummond, Tom Devine, Joep de Ligt, Sue Huang, Lindsay Lowe, Phil Shoemack, Lauren Jelley, Jemma L. Geoghegan, James Hadfield, Matt Storey, Andrew Sporle, Kip Mouldey, David Welch, Jillian Sherwood, Gerard J.B. Sonder, Colin R Simpson, Edward C. Holmes, G. Edwin Reynolds, Caroline McElnay, Tara Swadi, Xiaoyun Ren, Andrea McNeill, Sarah Jefferies, and Erasmus Smit
Subjects: Microbiology (medical), Aircraft, Isolation (health care), Genomic Evidence of In-Flight Transmission of SARS-CoV-2 Despite Predeparture Testing, Epidemiology, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), viruses, Physical Distancing, 030231 tropical medicine, United Arab Emirates, lcsh:Medicine, Disease, medicine.disease_cause, law.invention, 2019 novel coronavirus disease, lcsh:Infectious and parasitic diseases, respiratory infections, 03 medical and health sciences, 0302 clinical medicine, in-flight transmission, law, Quarantine, Humans, Medicine, lcsh:RC109-216, 030212 general & internal medicine, Coronavirus, business.industry, Transmission (medicine), SARS-CoV-2, Disease progression, lcsh:R, Masks, COVID-19, zoonoses, Infectious Diseases, coronavirus disease, Synopsis, business, Single mutation, New Zealand, Demography, severe acute respiratory syndrome coronavirus 2
Abstract: Since the first wave of coronavirus disease in March 2020, citizens and permanent residents returning to New Zealand have been required to undergo managed isolation and quarantine (MIQ) for 14 days and mandatory testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As of October 20, 2020, of 62,698 arrivals, testing of persons in MIQ had identified 215 cases of SARS-CoV-2 infection. Among 86 passengers on a flight from Dubai, United Arab Emirates, that arrived in New Zealand on September 29, test results were positive for 7 persons in MIQ. These passengers originated from 5 different countries before a layover in Dubai; 5 had negative predeparture SARS-CoV-2 test results. To assess possible points of infection, we analyzed information about their journeys, disease progression, and virus genomic data. All 7 SARS-CoV-2 genomes were genetically identical, except for a single mutation in 1 sample. Despite predeparture testing, multiple instances of in-flight SARS-CoV-2 transmission are likely.
Published: 2021

29. Abstract LB293: The evolution of MRD assays: Moving beyond the tumor-Informed bespoke NGS panel

Author: Paul Labrousse, Daniel Stetson, James Hadfield, Chris Abbosh, Ahmet Zehir, Carl Barrett, Brian Dougherty, and Darren Hodgson
Subjects: Cancer Research, Oncology
Abstract: The non-invasive detection of circulating tumor DNA (ctDNA) from plasma has been shown to have clinical value for detection of minimal residual disease (MRD), identification of emerging resistance to treatment, and predicting treatment response. The ability to detect MRD following curative treatment allows for the stratification of patients with higher risk of disease recurrence. Multiple studies, across hundreds of patients in different indications, including lung and colon cancer, have shown that tumor-informed personalized assays have high sensitivity to detect recurrence. The tumor-informed bespoke panels utilize sequencing of the tumor tissue to identify clonal somatic variants, which are then used to generate a personalized panel to track the variants in patient plasma. Next-generation MRD technologies are aiming to provide greater sensitivity by expanding methods beyond the small, personalized panels. Utilization of increased numbers of somatic variants is predicted to achieve a greater level of sensitivity for detection of MRD. Here we describe, C2I Genomics which provides a novel platform that exploits whole genome sequencing of the tumor tissue and plasma to define a somatic variant signature. An important difference of this method is that it develops a personalized bioinformatic signature rather than a personalized NGS panel. To understand the potential of this novel methodology, we utilized a set of 54 contrived clinical sample dilutions from three indications, with an expected (or engineered) VAF range of 0.2-0.002%, that have commutability to the intended use population. We found that C2I Genomics technology is highly sensitive, where for all returned samples, the assay routinely achieved detection down to the lowest dilution of 0.002% allelic frequency. Overall, the assay resulted in an average detection rate of 93% across all dilutions (n = 54). The aggregate detection rate of several tumor-informed assays, examining the same dilutions, yielded a detection rate of 68%. In summary, the C2I Genomics assay showed robust detection from contrived samples which are representative of clinical trial samples in both cfDNA yields and allele frequency. Citation Format: Paul Labrousse, Daniel Stetson, James Hadfield, Chris Abbosh, Ahmet Zehir, Carl Barrett, Brian Dougherty, Darren Hodgson. The evolution of MRD assays: Moving beyond the tumor-Informed bespoke NGS panel [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB293.
Published: 2023

30. From elimination to suppression: genomic epidemiology of a large Delta SARS-CoV-2 outbreak in Aotearoa New Zealand

Author: Lauren Jelley, Jordan Douglas, Xiaoyun Ren, David Winter, Andrea McNeill, Sue Huang, Nigel French, David Welch, James Hadfield, Joep de Ligt, and Jemma L Geoghegan
Abstract: New Zealand’s COVID-19 elimination strategy heavily relied on the use of genomics to inform contact tracing, linking cases to the border and to clusters during community outbreaks. In August 2021, New Zealand entered its second nationwide lockdown after the detection of a single community case with no immediately apparent epidemiological link to the border. This incursion resulted in the largest outbreak seen in New Zealand caused by the Delta Variant of Concern. Here we generated 3806 high quality SARS-CoV-2 genomes from cases reported in New Zealand between 17 August and 1 December 2021, representing 43% of reported cases. We detected wide geographical spread coupled with undetected community transmission, characterised by the apparent extinction and reappearance of genomically linked clusters. We also identified the emergence, and near replacement, of genomes possessing a 10-nucleotide frameshift deletion that caused the likely truncation of accessory protein ORF7a. By early October, New Zealand moved from elimination to suppression and the role of genomics changed markedly from being used to track and trace, towards population-level surveillance.
Published: 2022

31. RCandy: an R package for visualizing homologous recombinations in bacterial genomes

Author: Chrispin Chaguza, Gerry Tonkin-Hill, Stephanie W Lo, James Hadfield, Nicholas J Croucher, Simon R Harris, and Stephen D Bentley
Subjects: Statistics and Probability, Computational Mathematics, Computational Theory and Mathematics, Applications Notes, Molecular Biology, Biochemistry, Computer Science Applications
Abstract: Summary Homologous recombination is an important evolutionary process in bacteria and other prokaryotes, which increases genomic sequence diversity and can facilitate adaptation. Several methods and tools have been developed to detect genomic regions recently affected by recombination. Exploration and visualization of such recombination events can reveal valuable biological insights, but it remains challenging. Here, we present RCandy, a platform-independent R package for rapid, simple and flexible visualization of recombination events in bacterial genomes. Availability and implementation RCandy is an R package freely available for use under the MIT license. It is platform-independent and has been tested on Windows, Linux and MacOSX. The source code comes together with a detailed vignette available on GitHub at https://github.com/ChrispinChaguza/RCandy. Supplementary information Supplementary data are available at Bioinformatics online.
Published: 2021
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32. Fragmentation patterns and personalized sequencing of cell‐free DNA in urine and plasma of glioma patients

Author: Nitzan Rosenfeld, Jonathan C. M. Wan, Katrin Heider, Richard Mair, Marta Grzelak, Dominique-Laurent Couturier, Matthew D. Eldridge, Jing Su, Ymke van der Pol, Dineika Chandrananda, James Morris, Kevin M. Brindle, Colin Watts, Hui Zhao, Florent Mouliere, Irena Hudecova, Mareike Thompson, James Hadfield, Christopher Smith, Wendy N. Cooper, Davina Gale, Mouliere, Florent [0000-0001-7043-0514], Smith, Christopher G [0000-0001-7357-2737], van der Pol, Ymke [0000-0002-3040-5848], Chandrananda, Dineika [0000-0002-8834-9500], Cooper, Wendy [0000-0003-3416-9982], Gale, Davina [0000-0002-4521-8199], Brindle, Kevin [0000-0003-3883-6287], Rosenfeld, Nitzan [0000-0002-2825-4788], Mair, Richard [0000-0001-8235-5689], Apollo - University of Cambridge Repository, Pathology, CCA - Cancer biology and immunology, and CCA - Imaging and biomarkers
Subjects: Pathology, medicine.medical_specialty, Medicine (General), Urine, Cancer detection, QH426-470, Article, 03 medical and health sciences, Plasma, Cell-free Dna, 0302 clinical medicine, Cerebrospinal fluid, R5-920, Glioma, Genetics, Biomarkers, Tumor, Medicine, Humans, Fragmentation (cell biology), Liquid biopsy, cell‐free DNA, 030304 developmental biology, Cancer, Biomarkers & Diagnostic Imaging, circulating tumor DNA, 0303 health sciences, liquid biopsy, business.industry, fragmentomics, Sequence Analysis, DNA, Articles, medicine.disease, 3. Good health, EMBO03, gliomas, EMBO02, Cell-free fetal DNA, 030220 oncology & carcinogenesis, Mutation, Molecular Medicine, business, Cell-Free Nucleic Acids
Abstract: Glioma‐derived cell‐free DNA (cfDNA) is challenging to detect using liquid biopsy because quantities in body fluids are low. We determined the glioma‐derived DNA fraction in cerebrospinal fluid (CSF), plasma, and urine samples from patients using sequencing of personalized capture panels guided by analysis of matched tumor biopsies. By sequencing cfDNA across thousands of mutations, identified individually in each patient’s tumor, we detected tumor‐derived DNA in the majority of CSF (7/8), plasma (10/12), and urine samples (10/16), with a median tumor fraction of 6.4 × 10−3, 3.1 × 10−5, and 4.7 × 10−5, respectively. We identified a shift in the size distribution of tumor‐derived cfDNA fragments in these body fluids. We further analyzed cfDNA fragment sizes using whole‐genome sequencing, in urine samples from 35 glioma patients, 27 individuals with non‐malignant brain disorders, and 26 healthy individuals. cfDNA in urine of glioma patients was significantly more fragmented compared to urine from patients with non‐malignant brain disorders (P = 1.7 × 10−2) and healthy individuals (P = 5.2 × 10−9). Machine learning models integrating fragment length could differentiate urine samples from glioma patients (AUC = 0.80–0.91) suggesting possibilities for truly non‐invasive cancer detection., Gliomas are challenging to detect using tumor derived cell‐free DNA (cfDNA) in body fluids. In this study, two novel analysis methods (tumor‐guided sequencing and sWGS) were developed to explore the potential of using plasma and urine cfDNA as a liquid biopsy for this malignancy.
Published: 2021

33. Phylodynamics reveals the role of human travel and contact tracing in controlling the first wave of COVID-19 in four island nations

Author: Jemma L. Geoghegan, Matt Storey, Xiaoyun Ren, Christiaan Swanepoel, Cinthy L. Jiménez-Silva, Alexei J. Drummond, James Hadfield, Colin R Simpson, Joep de Ligt, Dong Xie, Fábio K. Mendes, David Welch, Jordan Douglas, and Remco R. Bouckaert
Subjects: 0301 basic medicine, Official statistics, medicine.medical_specialty, Coronavirus disease 2019 (COVID-19), Range (biology), coronavirus, Taiwan, Iceland, Context (language use), human movement, medicine.disease_cause, Microbiology, contact tracing, law.invention, 03 medical and health sciences, 0302 clinical medicine, law, Virology, Pandemic, Epidemiology, Added value, Regional science, medicine, AcademicSubjects/MED00860, 030212 general & internal medicine, Coronavirus, AcademicSubjects/SCI01130, AcademicSubjects/SCI02285, Australia, COVID-19, phylogenomics, 030104 developmental biology, Geography, Transmission (mechanics), Viral phylodynamics, Mobile phone, Contact tracing, Research Article, New Zealand, Demography
Abstract: BackgroundNew Zealand, Australia, Iceland, and Taiwan all saw success at controlling the first wave of the COVID-19 pandemic. As islands, they make excellent case studies for exploring the effects of international travel and human movement on the spread of COVID-19.MethodsWe employed a range of robust phylodynamic methods and genome subsampling strategies to infer the epidemiological history of SARS-CoV-2 in these four countries. We compared these results to transmission clusters identified by the New Zealand Ministry of Health by contract tracing strategies.FindingsWe estimated the effective reproduction number of COVID-19 as 1–1.4 during early stages of the pandemic, and show that it declined below 1 as human movement was restricted. We also showed that this disease was introduced many times into each country, and that introductions slowed down markedly following the reduction of international travel in mid March 2020. Finally, we confirmed that New Zealand transmission clusters identified via standard health surveillance strategies largely agree with those defined by genomic data.InterpretationWe have demonstrated how the use of genomic data and computational biology methods can assist health officials in characterising the epidemiology of viral epidemics, and for contact tracing.FundingThis research was funded by the Health Research Council of New Zealand, the Ministry of Business, Innovation, and Employment, the Royal Society of New Zealand, and the New Zealand Ministry of Health.Research in ContextEvidence before this studyOur study looks at the early months of the COVID-19 pandemic, a period in which the first wave was controlled in four “island” nations – New Zealand, Australia, Taiwan, and Iceland. All prior data used in this study was collected from late 2019 until the end of April 2020. This includes over 3000 SARS-CoV-2 genomic sequences which were collected in this period (and subsequently deposited into GISAID), as well as arrival and departure information (provided by official statistics from each country), human mobility data collected from mobile phones (by Apple), and COVID-19 case data (released by the World Health Organisation). Even early on during the COVID-19 pandemic, the properties of SARS-CoV-2 – including the reproduction number and mutation rate – were well characterised, and a range of these estimates have been covered in our article. Our Bayesian phylodynamic models, including their prior distributions, are informed by all of the above sources of information. Finally, we have incorporated all of the available information on COVID-19 transmission clusters identified by the New Zealand Ministry of Health during this period.Added value of this studyWe quantified the decline in the reproduction number of SARS-CoV-2, following the decline in human mobility, in four “island” countries. We also demonstrated how importation events of SARS-CoV-2 into each considered country declined markedly following the reduction of international travel. Our results shed a different light on these patterns because of (i) our locations of choice – the four countries had success in dealing with the first pandemic wave, with their geographic isolation contributing to cleaner signals of human mobility, and (ii) our novel and empirically driven phylodynamic model, which we built from explicitly modelling mobile phone data in the four islands. Furthermore, by crossing epidemiological against ge3nomic data, our paper quantitatively assesses the ability of contact tracing, as implemented by the New Zealand Ministry of Health (NZMH), in identifying COVID-19 transmission clusters. We find evidence for a high efficacy of the specific measures taken – and when they were taken – by the NZMH in identifying transmission clusters, considered worldwide to have been successful in its response to the pandemic. Our analyses also illustrate the power of viral genomic data in assisting contact tracing.Implications of all the available evidenceThe conclusions drawn from this research inform effective policy for locations pursuing an elimination strategy. We confirm the accuracy of standard contact tracing methods at identifying clusters and show how these methods are improved using genomic data. We demonstrate how the overseas introduction rates and domestic transmission rates of an infectious viral agent can be surveilled using genomic data, and the important role each plays in overall transmission. Specifically, we have quantified these processes for four countries and have shown that they did decline significantly following declines in human travel and mobility. The phylodynamic methods used in this work is shown to be robust and applicable to a range of scenarios where appropriate subsampling is used.
Published: 2021

34. RNA sequencing: the teenage years

Author: Marta Grzelak, Rory Stark, and James Hadfield
Subjects: Sequence analysis, genetic processes, Computational biology, Biology, History, 21st Century, Transcriptome, 03 medical and health sciences, 0302 clinical medicine, Transcription (biology), Gene expression, Genetics, Humans, natural sciences, RNA, Messenger, Nucleic acid structure, Molecular Biology, Genetics (clinical), 030304 developmental biology, 0303 health sciences, Sequence Analysis, RNA, Gene Expression Profiling, Alternative splicing, High-Throughput Nucleotide Sequencing, RNA, Alternative Splicing, RNA splicing, 030217 neurology & neurosurgery
Abstract: Over the past decade, RNA sequencing (RNA-seq) has become an indispensable tool for transcriptome-wide analysis of differential gene expression and differential splicing of mRNAs. However, as next-generation sequencing technologies have developed, so too has RNA-seq. Now, RNA-seq methods are available for studying many different aspects of RNA biology, including single-cell gene expression, translation (the translatome) and RNA structure (the structurome). Exciting new applications are being explored, such as spatial transcriptomics (spatialomics). Together with new long-read and direct RNA-seq technologies and better computational tools for data analysis, innovations in RNA-seq are contributing to a fuller understanding of RNA biology, from questions such as when and where transcription occurs to the folding and intermolecular interactions that govern RNA function.
Published: 2019

35. Stage 1 results of BrUOG 354: A randomized phase II trial of nivolumab alone or in combination with ipilimumab for people with ovarian and other extra-renal clear cell carcinomas (NCT03355976)

Author: Don S. Dizon, Katina Robison, Shannon Diane MacLaughlan David, Jason T Machan, Matthew James Hadfield, Eric I Marks, Rani Chudasama, Tarra Evans, Mary Lorraine Lopresti, Emma Safran, Michaela Kastura, Faith Hassinger, Ashlee Sturtevant, Roxanne Wood, Alexi A. Wright, Rochelle Strenger, Ursula A. Matulonis, Christina Bandera, Susana M. Campos, and Michael J. Birrer
Subjects: Cancer Research, Oncology
Abstract: 5598 Background: Clear Cell Carcinoma (CCC) outside the kidney is a rare tumor that can arise from multiple organs, including the ovary, endometrium and cervix. Extra-renal CCC is chemoresistant and has a poor prognosis. Data suggest that CCC of the gynecologic tract resembles the genomic profile of Renal Cell Carcinoma (RCC), which is responsive to immune checkpoint inhibition (ICI) therapy. We are conducting a two-stage phase 2 trial evaluating immunotherapy for extra-renal CCC. The primary objective is to assess overall rate of response (ORR); Progression-Free (PFS), Overall Survival (OS), and correlative biomarker studies are secondary. Here we present the results of Stage 1. Methods: This is a randomized two-stage non-comparative phase II study evaluating nivolumab (240mg IV every two weeks) alone (N) and in combination with ipilimumab (1mg/kg every six weeks, [N+I]) in patients with relapsed extra-renal CCC after at least one prior chemotherapy (no prior ICI), and measurable disease. Treatment was continued until disease progression or unacceptable toxicity. Stage 1 of this trial called for up to 30 volunteers (15 per arm) after which the study was closed. Consideration to reopen to stage 2 called for two or more responses in either arm. Here we present the completion of Stage 1; the release of results was approved by Brown University Oncology Group (BrUOG) Data Safety and Monitoring Committee. Results: Between July 2018 and October 2021, 30 patients were enrolled and 29 were treated (Table). The majority (83%) had CCC of the ovary (n=24). The ORR with N and N+I was 14.2 and 26.7%, respectively. The 6 month PFS rate was 19.1 and 43.8%; median PFS was 2.7 (95%CI 1.3-5.1) and 5.1 months (95%CI 0.9-NR), respectively. Grade ≥3 treatment-related toxicities occurred in 4 (28.6%) on N and 5 (33.3%) on N+I. There were no treatment-related deaths and no new safety signals. One volunteer enrolled on N+I stopped treatment after two years and remains in CR to date. Conclusions: Although sufficient activity was seen in CCC in both arms, the single-agent activity of N is similar to published reports in platinum-resistant epithelial ovarian cancer and decision made not to pursue it further. However, the combination of ipilimumab and nivolumab warrants additional investigation, and the second stage of this study will enroll 14 more patients to receive N+I. Clinical trial information: NCT03355976. [Table: see text]
Published: 2022

36. Real-time genomics to track COVID-19 post-elimination border incursions in Aotearoa New Zealand

Author: Jordan Douglas, Matthew Storey, Jemma L. Geoghegan, David Welch, Nigel P. French, Joep de Ligt, Xiaoyun Ren, James Hadfield, and Remco R. Bouckaert
Subjects: medicine.medical_specialty, 2019-20 coronavirus outbreak, Geography, Coronavirus disease 2019 (COVID-19), Public health, Genomic sequencing, Viral transmission, medicine, Outbreak, Genomics, Aotearoa, Genealogy
Abstract: There have been thirteen known COVID-19 community outbreaks in Aotearoa New Zealand since the virus was first eliminated in May 2020, two of which led to stay-at-home orders being issued by health officials. These outbreaks originated at the border; via isolating returnees, airline workers, and cargo vessels. With a public health system informed by real-time viral genomic sequencing which typically had complete genomes within 12 hours after a community-based positive COVID-19 test, every outbreak was well-contained with a total of 225 community cases, resulting in three deaths. Real-time genomics were essential for establishing links between cases when epidemiological data could not, and for identifying when concurrent outbreaks had different origins. By reconstructing the viral transmission history from genomic sequences, here we recount all thirteen community outbreaks and demonstrate how genomics played a vital role in containing them.SummaryThe authors recount the role of real-time viral genomics in containing the COVID-19 community outbreaks of Aotearoa New Zealand.
Published: 2021

37. Augur: a bioinformatics toolkit for phylogenetic analyses of human pathogens

Author: Emma B. Hodcroft, Misja Ilcisin, Elias Harkins, Richard A. Neher, Kairsten Fay, Trevor Bedford, Jover Lee, James Hadfield, John Huddleston, and Thomas R. Sibley
Subjects: Phylogenetic tree, Computer science, 610 Medicine & health, Human pathogen, Python (programming language), Bioinformatics, Genome, Article, Variety (cybernetics), ComputingMethodologies_PATTERNRECOGNITION, 360 Social problems & social services, Custom software, Perl, computer, computer.programming_language
Abstract: Summary and statement of need The analysis of human pathogens requires a diverse collection of bioinformatics tools. These tools include standard genomic and phylogenetic software and custom software developed to handle the relatively numerous and short genomes of viruses and bacteria. Researchers increasingly depend on the outputs of these tools to infer transmission dynamics of human diseases and make actionable recommendations to public health officials (Black et al., 2020; Gardy et al., 2015). In order to enable real-time analyses of pathogen evolution, bioinformatics tools must scale rapidly with the number of samples and be flexible enough to adapt to a variety of questions and organisms. To meet these needs, we developed Augur, a bioinformatics toolkit designed for phylogenetic analyses of human pathogens.
Published: 2021
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38. Timing of surgery following SARS-CoV-2 infection: an international prospective cohort study

Author: Dmitri, Nepogodiev, Joana Ff Simoes, Elizabeth, Li, Maria, Picciochi, James, C Glasbey, Glauco, Baiocchi, Ruth, Blanco-Colino, Daoud, Chaudhry, Ehab, Alameer, Kariem, El-Boghdadly, Funmilola, Wuraola, Dhruva, Ghosh, Rohan, R Gujjuri, Ewen, M Harrison, Herman, Lule, Haytham, Kaafarani, Mohammad, Khosravi, Irmgard, Kronberger, Sezai, Leventoğlu, Harvinder, Mann, Kenneth, A Mclean, Mengistu Gebreyohanes Mengesha, Maria Marta Modolo, Faustin, Ntirenganya, Lisa, Norman, Oumaima, Outani, Riinu, Pius, Peter, Pockney, Ahmad Uzair Qureshi, April Camilla Roslani, Sohei, Satoi, Catherine, Shaw, Aneel, Bhangu, Omar, M Omar, Waheed-Ul-Rahman, Ahmed, Leah, Argus, Alasdair, Ball, Edward, P Bywater, Amanpreet, Brar, Brett, E Dawson, Irani, Duran, Muhammed, Elhadi, Conor, S Jones, Sivesh, K Kamarajah, James, M Keatley, Samuel, Lawday, Ella, J Marson, Riinu, Ots, Irène, Santos, Elliott, H Taylor, Isobel, M Trout, Chris, Varghese, Mary, L Venn, William, Xu, Irida, Dajti, Arben, Gjata, Salah Eddine Oussama Kacimi, Luis, Boccalatte, Daniel, Cox, Philip, Townend, Felix, Aigner, Irmgard Elisabeth Kronberger, Elgun, Samadov, Amer, Alderazi, Kamral, Hossain, Greg, Padmore, Gabrielle van Ramshorst, Ismaïl, Lawani, Anis, Cerovac, Samir, Delibegovic, Gustavo Mendonça Ataíde Gomes, Igor Lima Buarque, Muhammad, Gohar, Mihail, Slavchev, Chukwuemeka, Nwegbu, Arnav, Agarwal, Janet, Martin, Joshua, Ng-Kamstra, Maricarmen, Olivos, Wenhui, Lou, Dong-Lin, Ren, Jose Andres Calvache, Carlos, J Perez Rivera, Ana Danic Hadzibegovic, Tomislav, Kopjar, Jakov, Mihanovic, Pablo Mijahil Avilés Jiménez, Nikolaos, Gouvas, Jaroslav, Klat, René, Novysedlák, Nicolas, Amisi, Peter, Christensen, Alaa, El-Hussuna, Sylvia, Batista, Eddy, Lincango-Naranjo, Sameh, Emile, Danilo Alfonso Arévalo Sandoval, Hailu, Dhufera, Samuel, Hailu, Mengistu, G Mengesha, Joonas, H Kauppila, Alexis, P Arnaud, Zaza, Demetrashvili, Markus, Albertsmeier, Hans, Lederhuber, Markus, W Löffler, Daniel Kwesi Acquah, Bernard, Ofori, Stephen, Tabiri, Symeon, Metallidis, Georgios, Tsoulfas, Maria-Lorena, Aguilera-Arevalo, Gustavo, Recinos, Tamás, Mersich, Dániel, Wettstein, Gabriele, Kembuan, Peiman Brouki Milan, Mohammad Hossein Khosravi, Masoud, Mozafari, Ahmed, Hilmi, Helen, Mohan, Oded, Zmora, Gaetano, Gallo, Francesco, Pata, Gianluca, Pellino, Yuki, Fujimoto, Naoto, Kuroda, Mohamad, K Abou Chaar, Faris, Ayasra, Ildar, Fakhradiyev, Intisar Hisham Said Hamdun, Jang, Jin-Young, Mohammad, Jamal, Lina, Karout, Aiste, Gulla, Fanjandrainy, Rasoaherinomenjanahary, Luc Hervé Samison, Iran Irani Durán Sánchez, Diana Samantha Gonzalez, Laura, Martinez, María José Martínez, Alejandra, Nayen, Antonio Ramos-De la Medina, Jade, Nunez, Pueya Rashid Nashidengo, Rakesh, Shah, Ashish Lal Shrestha, Pascal, Jonker, Schelto, Kruijff, Milou, Noltes, Pieter, Steinkamp, Deborah, Wright, Lukman, Abdur-Rahman, Adesoji, Ademuyiwa, Adewale, Adisa, Babatunde, Osinaike, Justina, Seyi-Olajide, Omolara, Williams, Emmanuel, Williams, Sofija, 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Mehraj, Ranganath, N, Ashish, Phadnis, Yadev, I, Alfie, Kavalakat, Rohin, Mittal, Karthik Chandra Vallam, Hamed, Akhavizadegan, Esmaeil Rezghi Maleki, Naser Yousefzadeh Kandevani, Hilary, Ikele, Catherine, Mcnestry, Christina, Fleming, Stephen, O'Brien, Sami Abd Elwahab, Niall, Davis, Mohsen, Javadpour, Brendan, Mcdonnell, Clare, O Connor, Jarlath, Bolger, Cillian, Clancy, Stefanie, M Croghan, Noel, Donlon, Carolyn, Cullinane, Ben, Creavin, Muheilan, Muheilan, Helen, Earley, Syed Mohammad Umar Kabir, Muhammad, Fahadullah, Éanna, Ryan, Tara, Connelly, Daisuke, Hashimoto, Majdi Ali Alqudah, Amer, Alajalen, Rand, Y Omari, Abdulrahman, Qasem, Yazan, Alawneh, Amer, Ahmad, Omar, Aladawi, Bourhan, Alrayes, Hanan, Haidar, Shatha, Husain, Faisal, Qassem, Adnan, Sumadi, Ala'a Abu Salhiyeh, Balqees Mahmoud Al-Manaseer, Zaid, Alsunna, Hazim, Ra'Ed, Faten Reyad Bani Hamad, Amro, Abuleil, Elmi Ahmed Mohamed Jimaale, Marah, Abu-Mehsen, Noor, Olaywah, Omar, Wafi, Hazim, Ababneh, Luai, Abu-Ismail, Almu'Atasim, Khamees, Ahmad, Alkhatib, Raikhan, Bolatbekova, Mukhtar, Kulimbet, Talgat, Nurgozhin, Timur, Saliev, Baurzhan, Zhussupov, Ydyrys, Almabayev, Dilyara, Kaidarova, Khalil, Tamoos, Ahmed, Aqeelah, Alsnosy Abdullah Khalefa Mohammed, Faraj Al Maadany, Ghadah, Alkadeeki, Milad, Gahwagi, Wafa, Aldressi, Mohamed, Amnaina, Arowa Hassan Abdulrahman Alansari, Akram, Alkaseek, Ghozlan, Yagoub, Anass Ben Amer, Marwa, Salem, Ayman, Almugaddami, Dania, Burgan, Mohammed, Abdelkabir, Khayriyah, Alshareef, Rayet Al Islam Ben Jouira, Ayman, Meelad, Ahmad, Bouhuwaish, Sumayya Essayah Dwaga, Houda, Khalifa, Bushray, Almiqlash, Taha, Suliaman, Mohammed, Alawami, Fras, Elhajdawe, Hajir, Aboazamazem, Ibrahim, Ellojli, Ahmed, Msherghi, Ismail Ali Saleh, Mohammed, Alayan, Marcel Didier Ndayishyigikiye, Akutu, Munyika, Philipp, Plarre, David, W Borowski, Cameron, Wells, Rebecca, Teague, Brodie, Elliott, David, Kieser, Omar, Mohyieldin, Nick, Mcintosh, Cheyaanthan, Haran, Jasmin, King, Jeong, Ha, 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Débora Schalge Campioto, Mayra Tuboi Lamonica, Gustavo, Urbano, Ricardo Villela Prado, Nivaldo, Alonso, Carlos Ferreira Dos Santos, Cristiano, Tonello, Robinson Esteves Pires, Ricardo Fernandes Rezende, Igor, Reis, Fernando Augusto Lima Marson, Erick Pires Ferreira, Camila Vantini Capasso Palamim, Marcelo, Brandao, Tiago Henrique de Souza, Rebecca, Maunsell, Alfeu Accorsi Neto, Guilherme, Accorsi, Murilo Francisco Fernandes, Camila, Machareth, Barbara Viegas Moura, Vanessa, Dias, Luciano, Guarienti, Lia Regina de Sampaio, Cristiano, Vendrame, Tatiane Amorim Coelho, Karin, Becker, Andre, Dias, Camila Girardi Fachin, Monica Maria Gomes-da-Silva, Isabela, Moraes, Amanda, Pinto, Adriano Seikiti Stychnicki, Nathalia Siqueira Julio, José Mauro Dos Santos, Humberto Fenner Lyra Junior, João Carlos Costa de Oliveira, Tiago Rafael Onzi, Marlus Tavares Gerber, Filipe Osni Coelho, Janaína Carla da Silva, Carolina, Panis, Daniel, Rech, Melissa, Avelino, Lais, Botacin, Mateus Capuzzo Gonçalves, Amanda Lira Dos Santos Leite, Laercio, Pol-Fachin, Tainá Santos Bezerra, Alcimar Lavareda Dos Santos Junior Alcimar, Robson, Amorim, Maria Eduarda Bellotti Leão, João José Corrêa Bergamasco, Cintia Cardoso Pinheiro, Rubem Alves Da Silva Neto, Elaine Francisca De Araújo, Lilian, Guimaraes, Tatiana, Liborio-Kimura, Isabelle Melo da Camara, Victor Ripardo Siqueira, Juan, Rodriguez, Jeancarllo, Silva, Rubem Alves Silva Junior, Cláudio, Cardoso, Andre, Guimaraes, Wislene Sarajane Moreira Alves, Agna Soares Da Silva Menezes, Marcelo, Araujo, Kattiucy, Brito, Josie Marcelle Lira Albuquerque, Jairo Alberto Dussan-Sarria, Andressa, Souza, Andre Ricardo Stüker, Tiago, Bresciani, Luciana Cadore Stefani, Leandro Totti Cavazzola, Tainá, Costa, Matheus, Dasqueve, Gustavo De Bacco Marangon, Otávio Ritter Silveira Martins, Josy, Rodrigues, Guilherme Roloff Cardoso, Brasil Silva Neto, Tilaê, Soares, Aline, Zanella, Lucas Torelly Filippi, Enilde Eloena Guerra, Ricardo Pedrini Cruz, Antonio Nocchi Kalil, Gustavo, Laporte, Moacyr, Salem, Murilo de Lima Brazan, Caio Antonio de Campos Prado, Elaine Christine Dantas Moises, Hilda Satie Suto, Alice Gadotti Yasuda, Ana Carolina Tagliatti Zani, Roberto Cardoso Cardoso Dos Santos, Geraldo, Duarte, Edwaldo Edner Joviliano, Carolina Lourenço Gomes Dos Santos, Silvana Maria Quintana, Edwin, Tamashiro, Fabiana Cardoso Pereira Valera, Marília Veccechi Bijos Zaccaro, Isis, Andreotti, Leonardo, Lima, Wilson Salgado Jr, Alonço da Cunha Viana Júnior, Lana, Moutinho, Cora Pichler de Oliveira, Isabela Trindade Martins, Isabela Vieira Toledo, Marco Antonio Correa Guimaraes-Filho, Rodrigo, Arrivabeno, Claudio Bovolenta Murta, Danniel Frade Said, Jose Pontes Junior, Felipe Guimarães Pugliesi, Paulo, Gregorio, Alessandro, Mariani, Fabio, Minamoto, Juliana Lourenço da Silva, Bruno, Muller, Caio, Zanon, Vladimir, C Carvalho, Ana Lucia Munhoz Lima, Priscila, R Oliveira, Jorge Dos Santos Silva, Mariana Faccini Teixeira, Marco, Kulcsar, Leandro, Matos, Kamilla Schmitz Nunes, Joel Abdala Junior, Emne, Abdallah, Genival Barbosa Carvalho, Igor Correia de Farias, Cassia da Silva, Fabio Fernando Eloi Pinto, Luciana, Facure, André, Godoy, Bruna, Kupper, Fernanda, Leite, Matheus de Melo Lôbo, Henrique, Mantoan, Narimã, Marques, Tomas, Marques, João Paulo Medici, Silvio Melo Torres, Katheryne Merlos Garcia, Rafael Ribeiro Meduna, Ana Carolina Scintini Herbst, Silvana Soares Dos Santos, Bruna Tirapelli Gonçalves, Marcelo, Antonini, Carolina, Brienze, José Francisco Farah, Rodrigo, Gonçalves, Vinicius, Machado, Luis Roberto Nadal, Danilo Nadal Rodrigues, Adrieli, Pansani, Maria Luiza Rocha, Marcelo, Simonsen, Daniela, Tsuchiya, Rafaela, Vasques, Jorge, Amorim, Ana Alyra Carvalho, Rebeca, Correia, Brena Costa Dos Santos, Henrique Jorge Guedes Neto, Luis, Nakano, Mariana, Pereda, Vladimir, Vasconcelos, Jamile Barakat Awada, Guilherme, Gava, Tiago Riuji Ijichi, Nam Jin Kim, Rafael, Nunes, Fabio, Pinto, Thiago Henrique Sigoli Pereira, Gustavo Jardim Volpe, Arthur, Gatti, Caroline, Nardi, Ramon, Oliva, Layze Braz de Oliveira, Herica Emilia Félix de Carvalho, Ivonizete Pires Ribeiro, Gabriel Renan Soares Rodrigues, Álvaro Francisco Lopes de Sousa, Michel, Chebel, Paulo Henrique de Sousa Fernandes, Marcelo Augusto Faria Freitas, Juliano Rodrigues da Cunha, Zeenia, Ather, Dobromir, Dimitrov, Emil, Filipov, Elitsa, Gyokova, Ivelina, Ilieva, Tsvetomir, Ivanov, Martin, Karamanliev, Vasil, Neykov, Boyko, Atanasov, Nikolay, Belev, Paolina, Kamenova, Kaloyan, Tonev, Tsanko, Yotsov, Evguenia, Hristova, Kolyo, Spassov, Tsanko, Tsankov, Dragomir, Dardanov, Petar, Gribnev, Manol, Sokolov, James, Brown, John, Tanyi, Heather, Hurdle, Anthony, Maclean, Michael, Sander, Salim Al Riyami, Krittika, Bali, David, Bigam, Khaled, Dajani, Mehran, Anvari, Rafik, Bolis, David, Choi, Susan, Ellis, Michael, Gupta, Wael, Hanna, Dennis, Hong, Cynthia, Horner, Lea, Luketic, Peter, Moisiuk, Yaron, Shargall, Hilda, Alfaro, Nawar, Alkhamesi, Laura, Allen, Muriel, Brackstone, Eunice, Chan, Davy, Cheng, Jason, Chui, Nelson, Gonzalez, Brent, Lanting, S Danielle MacNeil, Robert, Mayer, Jacob, Mcgee, Mahesh, Nagappa, Nicholas, Power, Agya, Prempeh, Mehdi, Qiabi, Hassan, Razvi, Emil, Schemitsch, Herman, Sehmbi, Ushma, Shah, Yamini, Subramani, Edward, Vasarhelyi, Kelly, Vogt, Homer, Yang, Ali Ghorbani Abdehgah, Saba, Balvardi, Liane, Feldman, Julio Flavio Fiore Jr, Melissa, Hanson, Brent, Hopkins, Pepa, Kaneva, Lawrence, Lee, Julia, Leonard, Jad, Abou-Khalil, Andre, Martel, James, Stevenson, Susan, Lee, Richard, Merchant, Samuel, Avoine, Laurence, Belanger, Mathieu, Belanger, Etienne, Belzile, Anne-Sophie, Blais, Sofia, Boucher-Kovalik, Cindy, Boulanger-Gobeil, Etienne, Cardinal, Pierre-Olivier, Champagne, Jonathan, Cloutier, Simon, Corriveau-Durand, Mathieu, Cote, Maxime, Cote, Mathilde, Côté, Valérie, Courval, Suzanne, Demers, Christine, Desbiens, Mehdi El Ouazzani, Karine, Girard, Ève-Marie, Girard, Karo, Gosselin, Helene, Khuong, Nathalie, Labrecque, Eve-Lyne, Langlais, Audrey, Larouche, Andréane, Lavallée, Madeleine, Lemyre, Sarah, Maheux-Lacroix, Melissa, Marien, Patrick, Marin, Julie, Mauger, Genevieve, Milot, Sylvie, Nadeau, Sebastien, Nguyen, Mélinda, Paris, Stéphane, Pelet, Marie, Plante, Carole, Plante, Frederic, Pouliot, Marie Claude Renaud, Mathilde, Sarlabous, Isabelle, Schmit, Raphaël, St-Germain, Brian, Johnston, Nicole, Labine, Amit, Persad, Hong, Pham, Melissa, Wood, Riordan, Azam, Amit, Atrey, Andrew, Beckett, Daniel, Cohen, Sunit, Das, Julian, Daza, Cheryl, Dunkerton, Yosef, Ellenbogen, Ciara, Hanley, Sorcha, Kellett, Amir, Khoshbin, Karim, Ladha, Amanda, Mcfarlan, Spencer, Montgomery, Janneth, Pazmino-Canizares, Hrishikesh, Suresh, Ravleen, Vasdev, Maira, Ahmed, Fred, Gentili, Aristotelis, Kalyvas, Matthias, Millesi, Can, Sarica, Taariq, Shaikh, Leslie, St, Jacques, Raha, Tabasinejad, Gelareh, Zadeh, Marvin, Hsiao, Nicole, Jedrzejko, Peter, Black, Claire, Broe, Charlotte, Dandurand, Rebecca, Grey, Philemon, Leung, Andrew, Lindberg, Juan Mata Gutierrez, Kelly, Mayson, Adam, Meneghetti, Drew, Phillips, John, Street, Carolina Carvajal Calderón, Daniel Alejandro Donoso Pizarro, Esteban, Fernández, Ángela, Molero, Nastassja, Mutarello, Jorge Núñez Lucic, Katherine Ochoa Gaete, Catalina Ruiz Lopez, Rina, Sepúlveda, Camila, Ulloa, Mauricio, Barreda, Matias, Dallaserra, Matías Günther Wood, Mauricio Sandoval Tobar, Francisco, Garcia-Huidobro, Jimena, Dona, Carolina Soto Diez, Daniel, Igor, Javier, Mena, José Tomás Reyes, Catalina Arredondo Soto, Jose, Campos, Javier, Gonzalez, Barbara, Rivera, Jaime, Altamirano-Villarroel, David, Cohn, Luis Marin de Amesti, Pedro, Recabal, Camilo, Sandoval, Bruno Catoia Fonseca, Monica, Contador, Andres, Hodali, Camila, Pincheira, Andrea Ramos Mantilla, Claudio Salas Garrido, Janina, Torres, Marco, Valenzuela, Zehua, Chen, Jiankun, Hu, Jin, Tao, Kun, Yang, Yuexin, Zhang, Yuan, Chen, Wang, Cunchuan, Wah, Yang, Xueli, Bai, Tingbo, Liang, Tao, Ma, Ning, Pu, Hanlin, Yin, Camilo, Caicedo, Diego Felipe Tellez Beltran, Alejandro, Escobar, Carolina Perez Granados, Lisbeth Alexandra Urueña Pinzon, Carlos, Bonilla, Diana, Santana, Oscar, Serrano, Andrea, Garcia, Fernando Giron Luque, Nasly, G Patino-Jaramillo, Laura Giselle Contreras Baquero, Nestor Augusto Muñoz Botero, Jhoana Andrea Murillo Castellanos, Andrea Juliana Vega Calvera, Lina, M Acosta Buitrago, Maria Paz Bohórquez-Tarazona, Paulo Andrés Cabrera Rivera, J Felipe Casas, Valeria Cormane Alfaro, Julian, Corso, Bayron, Guerra, Albert Franz Guerrero-Becerra, Akram Kadamani Abiyomaa, Felix Ramon Montes, Manuel Santiago Mosquera Paz, Camilo Andrés Polanía Sandoval, Natalia, Quintana, Carlos Fernando Roman Ortega, Carlos, Santacruz, Manuela, Téllez, Camilo Alejandro Velandia Sánchez, Fernando, Arias-Amézquita, Luis Felipe Cabrera Vargas, Natalia Cortes Murgueitio, Elkin, Escorcia, Kemel Ahmed Ghotme Ghotme, Jorge Luis Gomez-Mayorga, Alvaro Felipe Guerrero Vergel, Gabriel, Herrera-Almario, Eduardo, Londono-Schimmer, Juan, A Mejia, Guillermo, Monsalve, Camilo, Rodriguez, Juan Nicolas Rodriguez Niño, Luis Martín Rodríguez Ortegón, Daniel, Sanabria, Andres, Alvarez, Arnulfo, Andrade, Javier, Ardila-Montealegre, David, Baquero, Edgar Mauricio Barrios Vidales, María Alejandra Caicedo Giraldo, María Camila Carvajal, Maria, Castillo, Maria Carolina Castillo Florez, María Angelica Cendales, Danny Conde Monroy, Henry, Cortes, Mario, Daniel, Jairo De la Peña, Elena Leonor Delgado-Nieto, Hernando, Espitia, Carlos Figueroa Avendaño, Edgar Oswaldo Hernandez Burgos, Andres, Isaza-Restrepo, German, Londono, Margarita Maria Maldonado, Andrea, Montenegro, Juan Carlos Navarro, Jorge, Navarro-Alean, Katherine Parra Abaunza, Eliana, Pulido, Erika, M Ramírez Amaya, Carlos Eduardo Rey Chaves, Natalia Andrea Rivera Rincón, William Mauricio Riveros Castillo, Lizeth Rodriguez Sanchez, Wilson, Rubiano, Maria, Russi, Juan Carlos Sabogal Olarte, Javier Mauricio Salgado Tovar, Maria Juliana Sanchez, David Mauricio Solano Varela, Paula Torres Gomez, Marcial, Trillos, Ana María Vargas Patiño, Felipe, Vargas-Barato, Juan Pablo Villate Leon, María Alejandra Wagner Useche, Jesus, Acosta, Adriana, Almeciga, Luis Becerra Mendez, Rafael Jose Beltran, Ana, Bonilla, Miguel, Buitrago, Marino, Cabrera, Pedro Hernando Calderon Quiroz, Diego, Camacho-Nieto, Carlos Andrés Carvajal Fierro, Sergio Cervera Bonilla, Sandra, Diaz, Helena, Facundo, Jorge, Forero, Mauricio Garcia Mora, Karena Victoria Garcia Tirado, German Fabian Godoy Perez, Felipe, Gonzalez, Oscar, Guevara, Jairo Alonso Hernandez, David Ricardo Herrera Mora, Juan David Lalinde, Carlos, Lehmann, Eduardo Leon Llanos, Byron Eduardo Lopez De Mesa Lopez, Ivan, Mariño, Monica, Medina, Raúl Eduardo Pinilla Morales, Angela, Puerto, Andrea, Quintero-Ortíz, Juliana Rendón Hernández, Elio Fabio Sánchez Cortés, Alvaro Eduardo Sánchez Hernández, Raul, Suarez, Oscar, Suescun, Marco, Vanegas, Rodolfo, Varela, Jorge Luis Velez Bernal, David, Viveros-Carreño, Daniela Camargo Gómez, Angelica, Fletcher, Abel, Merchan, Euler Javier Burbano Luna, Indira, Cujiño, Mauricio Velasquez Galvis, Lina Maria Vergara Galliadi, Orlando Abonia Gonzalez Abonia Gonzalez, Maria Fernanda Acuna Saravia, Roberto, Arroyave, Jose Maria Barreto Angulo, Francisco Javier Bonilla-Escobar, Uriel, Cardona, Ivan Castañeda Giacometto, José Luis Castillo, Diego Fernando Castillo-Cobaleda, Diego Jose Caycedo Garcia, Brenda Marcela Coll Tello, Juan Carlos Dueñas-Ramirez, Oscar Andres Escobar Vidarte, Luis, Figueroa, Herney, Garcia-Perdomo, Alden, Gomez, Laura, Gomez, Alejandro, Gomez, Ana María Grande-Gil, David, Guarin, Natalia, Guzman, Enrique Herrera Castañeda, Marisol, Hinaoui, Anuar Armando Idrobo Escobar, Christian Kammerer Kammerer, Sergio, Leon, Carlos Antonio Llanos Lucero, Angie, López, Alexander Maximiliano Martinez-Blanco, Antonio José Montoya Casella, Ricardo Andres Niño Corredor, Sebastian, Ordoñez, Diego Alfredo Palta Uribe, Andrea Carolina Perea Serna, July, Ríos, Gabriel, Ríos-Samper, Juan David Rivera Garcia, Juan Camilo Salcedo Moreno, John, Sandoval, Guillermo Alberto Sarmiento Ramirez, Paola Andrea Tabares Romero, Ricardo, Urzola, Iliana Maria Valdes-Duque, Miguel, Velasquez, Lina Maria Villegas, James, Zapata-Copete, José Omar Zorrilla Lara, Mauricio Zuluaga Zuluaga, Alejandra Echeverri Moreno, Juan Sebastian Figueroa, Adolfo Enrique Gómez Ortiz, Maria Fernanda Gonzalez Mosos, Alejandro, González-Orozco, Juan Jose Jaramillo Roncancio, María Camila Leyva Martínez, Monica, Mosos, Juan Sebastian Ramirez, Henry Andrés Rodríguez, Juan David Saavedra Henao, María Alejandra Torrado Varón, Leidy Natalia Idarraga Ramírez, Lorena, Ocampo, Monica Cardona Marin, Ana Maria Marin Gonzalez, David Alejandro Mejia, Luis Emiro Vanegas, Yicel Alvarez Martinez, Sandra Aruachan Vesga, Dayana Vanessa Baron Fuentes, Alejandra Caballero Salas, Gustavo Antonio Martinez Estrada, Kelly Viloria Campo, Juan Antonio Corralez Alvarez, Cesar David Galindo Regino, Angela Maria Giraldo Velasquez, Carlos Enrique Melo Moreno, Jorman, H Tejada, Jesus Hernan Tovar, German Alirio Tovar, Edison Alexander Benavides Hernández, Claudia Milena Orozco-Chamorro, Gustavo Adolfo Angel, Christian Ali Buesaquillo, Victor David Olave Montaño, Andrés, Sánchez-Gómez, Daniela Patricia Escalante Ureche, Jairo Martinez Garrido, Eben-Ezer, Genda, Fabrice, Eboma, Kristina, Bitunjac, Karlo, Grulović, Marijana, Vučković, Goran, Šantak, Emanuel, Borovic, Ana Bosak Versic, Damir, Hasandić, Natasa Poldan Grabar, Suzana Srsen Medancic, Lucija, Brkic, Sara, Cokarić, Petra Pavic Palac, Zeljka, Samac, Josipa, Tomić, Marija, Vrdoljak, Ivan, Bacic, Bernarda, Bakmaz, Nikolina Bratošević Vučičić, Domagoj, Brzic, Samir, Canovic, Emilio, Dijan, Maja Grgec Dragicevic, Robert, Karlo, Suzana, Konjevoda, Petra, Kovačević, Ivan, Kovačić, Frane, Markulić, Luka, Matak, Domagoj, Morović, Gordan, Perišić, Andrea, Simic, Neven, Skitarelić, Matea, Veršić, Vanja, Žufić, Matea, Zuzul, Srdan Ante Anzic, Iva, Carevic, Dubravka Heli Litvic, Duska, Markov-Glavas, Tatjana Savic Jovanovic, Goran, Augustin, Jerko, Biloš, Bojan, Biočina, Vedrana, Biosic, Dino, Bobovec, Iva, Botica, Boris, Bumber, Petra, Čerina, Katarina Duric Vukovic, Hrvoje, Gasparovic, Lucija, Gatin, Kresimir, Grsic, Ika Gugić Radojković, Ivan, Jelčić, Zeljko, Kastelan, Juraj, Kolak, Tomislav, Kulis, Kristian, Kunjko, Marjan, Maric, Marcel Marjanović Kavanagh, Borna, Milicic, Trpimir, Morić, Miram, Pasini, Luka, Penezić, Drago, Prgomet, Ratko, Prstacic, Andreja, Prtorić, Rudolf, Radojković, Ivan, Romić, Tomislav, Sečan, Dora Škrljak Šoša, Juraj, Slipac, Mislav, Tomic, Jurica, Zedelj, Toni, Zekulić, Zoran, Zimak, Mia, Lorencin, Ivica, Luksic, Matija, Mamic, Norberto Miranda Espinsa, Ailén Sánchez Cruz, Stavros, A Antoniou, Michael, Kakas, Yiannis, Panayiotou, Heyam, Almezghwi, Kalbim, Arslan, Hasan, Besim, Ali, Özant, Necdet, Özçay, Georgios, Kokkinos, Panayiotis, Papatheodorou, Ioanna, Pozotou, Olga, Stavrinidou, Anneza, Yiallourou, Lukáš, Burda, Michal, Dosoudil, Lubomir, Martinek, Wladyslaw, B Gawel, Milan, Lerch, Matúš, Peteja, Jan, Žatecký, Toman, Daniel, Martin, Formánek, Veronika, Javurkova, Markéta, Kepičová, Nicole, Macečková, Petr, Matousek, Ondrej, Simetka, Petr, Vavra, Karol, Zelenik, Jan, Černý, Sami, Khan, Robert, Lischke, Martin, Přibyl, Temoore, Younus, Yenuksha, Amarasena, Karel, Klíma, Setareh, Pirmorad, Zuzana, Hotová, Aleš, Mladěnka, Nulvin, Bozo, Julie Lykke Harbjerg, Louise, Hviid, Helle, Ø Kristensen, Mira, Mekhael, Mette Fugleberg Nielsen, Laerke, Paulsen, Saija, Sinimäki, Louise Zinck Mogensen, Peter, Bonde, Anders Lyng Ebbehøj, Anne-Sophie, Fenger, Aleksander Fjeld Haugstvedt, Christine Hangaard Hansen, Maria Lovisa Jönsson, Lars, N Jorgensen, Peter-Martin, Krarup, Anne-Louise, Lihn, Christian, Meyhoff, Helena, Otte, Anas Ould Si Amar, Henrik, Palm, Nis, Schlesinger, Henry, Smith, Ida, Tryggedsson, Anne Reiss Axelsen, Jens Kristian Baelum, Mark Bremholm Ellebaek, Signe Bremholm Ellebaek, Anders, Hogh, Maria, Acosta, Yancy, Acosta, Pedro, Baez, Luis Alfredo Betances, Luis Rodolfo Bonilla, Daniel, Cabreja, Wilton Cabrera Cruz, María Mabel Collado Expósito, Aldo, Crespo, Octavio, Cruz-Pineda, Claudio Samuel D'óleo García, Pedro, Díaz, Remberto, Escoto, Jatnna, Figueroa, Luis, Garcia, U Garcia-Dubus Rodriguez, Humberto, Gomez, Adrian, Grullon, Fatherin, Guerrero, Lillian, Guzman, Thelma Rocío Jiménez Mosquea, Engels, Lazala, Ariela, Lopez, Leeany Maletta Francisco, Jhomayri, Mercado, Herisardy, Munoz, Jeffrey, Paulino, Maricely Ambar Perez Fernandez, Rudeily, Reyes, Lilia Rosa Reyes Guilamo, Ruben, Rivas, Claudia Alejandra Rivas Torres, Ada, Rodríguez, Madelin Tamar Rosario Villa, Eleazar, Santana, Joel, Soto, Nassin, Tactuk, Cristina Kristel Tonos Sardiñas, Raul, Ubinas, Jose, Victoria, Aaron, Villegas, Ricardo, Acra-Tolari, Larissa, Beltran, Luis Fernand Betances, Alejandro, Blaubach, Denisse Idalia Campos Mejía, Nathalia, Capellan, María Fernanda Cedeño Bruzual, Saray Cordero Spencer, Gabriella Cuevas Lantigua, Luis De Jesus, Gabriela, Díaz, Pedro Pablo Díaz Vásquez, Karla, Disla, Jesús Antonio Echavarría Uceta, Paola Irina Eusebio Jimenez, Damaris, Fernandez, Marlin Fernandez Camilo, Jiomar Manuel Figueroa Germosen, Maruel, Fortunato, Leyla Gabriel Fernández, Alicia German Dihmes, Héctor, Herrera, Elizabeth Leon Cuevas, Jose Alejandro Mata, Mirian Mateo De La Cruz, Dolores Mejia De la Cruz, Marcos, Mirambeaux, Fabio Ortiz De La Cruz, Henry, Paulino, Gabriela, Pelletier, Merycarla, Pichardo, Rodriguez Pumarol Próspero Enrique, Julio, Rivas, Ann Stephany Sanchez Marmolejos, Roman Santana Santana, Irina Suero Almanzar, María Magdalena Vásquez Sánchez, Nicolás, Campuzano, José Ricardo Negrete Ocampo, Maria, Armas, Alvaro Santiago LeMarie Guerra, Gustavo León Vizcaya, Omar Ahmed Abdelwahab, Ahmed, O Elmehrath, Mohammed Ezzat Mostafa, Gamal, Amira, Ibrahim, Sallam, Mohamed, Sherief, Ahmed, Abdelmajeed, Mostafa, Abdou, Ahmed Abo Shanab, Aya, Abodeeb, Roger, Aboelkhel, Nour Eldin Abosamak, Amna Abou Bakr, Samar, Aboubakr, Yossof, Abouelnagah, Omar, Aboumadawy, Dina, Adel, Nermeen, Afifi, Sara, Ahmed, Abedelrahman, Ahmed, Mohamed Al Sayed, Abdelrahman, Alberkamy, Mohamed, Aldahma, Ahmed, Ali, Mostafa, Ali, Mohamed Amin Bakr, Alaa, Anter, Doaa, Asal, Fouad, Ashoush, Olfat, Ashraf, Abdel Rahman Ashraf, Rewan, Atta, Salma, Badr, Mohamed, Bahaaeldin, Mostafa, Bastawesy, Sara, Darwish, Rasha El Kharashy, Ziad, Elassar, Seifeldin, Elbadawy, Dina, Elkhity, Alromisaa, Elsaka, Manal, Elsayed, Mostafa Elsayed Elsayed Hewalla, Mohamed, Elshafey, Youssof, Eshac, Fares, Eshac, Amr, Essameldin, Moataz, Ewedah, Yara, Ezz, Mohamed, Farag, Inas, Gadelkarim, Ziyad, Gadelrab, Dina, Gamal, Mira, Ghaly, Nathalie, Girgis, Omar, Gouda, Nancy, H El Goweini, Nour, Hafez, Youssef, Hafez, Abdelkader Hamed Abdin, Rodina, Hanno, Aliaa, Hassanin, Mohamed, Hassanin, Hanan, M Hemead, Abdelrahman, Hemida, Maher, Hosain, Ahmed, Hossam, Hamza Hussein Aly Salama Aly, Mohamed, Ibrahim, Abdelrahman, Ibrahim, Nourhan, Ibrahim, Noha Mohamed Salah Ibrahim Moussa Hamouda, Monica, Iskander, Islam, Khalifa, Mostafa, Kotb, Nada, Mahmoud, Alaa Mahmoud Abo Shabana, Nora, Mamdouh, Maram, Metwalli, Khaled, Metwally, Mahmoud, Moghazy, Moustafa, Mohamed, Mohamed, Mourad, Esraa, Moustafa, Kamilia, Mubarek, Nour Eldin Nader, Ahmed, Nasser, Samaa, Omar, Mostafa Shehata Qatora, Mohamed, Ragab, Marawan, Ragal, Dina, Ramadan, Rana, Ramadan, Sara, Ramadan, John, Romany, Ahmed, Sabry, Yasmeen, Said, Sara, Salamah, Ahmed Mostafa Saleh, Mona Saleh Mesbah Mohamed Elkaffas, Alaa, Salem, Hashem, Salim, Ahmed, Samih, Ahmed Samir Abdelaal, Ahmed, Shaheen, Yassmeen Sharafeldin Mohammed, Abdelrahman, Shehata, Mohamed, Shemeis, Karim, Shenit, Mennatallah, Sheta, Asmaa, Soffar, Yousef, Tanas, Ahmed, Tarek, Nermin, Yehia, Omar Khaled Mohamed Eid, Ammar Yasser Abdulfattah, Ahmed, Elsaghir, Mohamed Fouad Elganainy, Omar Ibrahim Elsayed, Omar, Elmandouh, Omar, Hamam, Mostafa Ahmed Shehata, Areej, A Abdelaziz, Ahmed, M Abbas, Wael Abd El-Ghani, Hossam Aldein, S Abd Elazeem, Mustafa Abd Elsayed, Ahmed Yassien Abd-Elkariem, Shimaa, Abdalla, Mahmoud, Abdel-Aleem, Khaled, Abdelazeem, Mahmoud, Abdelfattah, Mohammed, Abdelhafez, Mohamed, M Abdelkarem, Ali, Abdelraouf, Lamess, Abdullaha, Hossam, Abubeih, Moaiad Eldin Ahmed, Nagm Eldin Abu Elnga Ahmed, Ahmed, Ahmed, Sarah Ahmed Saad, Sherif, Alaa, Ali, Alhussaini, Abdelrahman Ahmed Abdelrahman Ali, Ibrahim, Ali, Wagdi, Ali, Mohamed Ali Mohamed, Mohamed, Ashraf, Muhammad, Bassiouni, Mohamed, G El-Adawy, Mohammad, El-Sharkawi, Hussein, Elkhayat, Khaled, Elmaghraby, Shady, Elsdfy, Mohamed, Elsharkawy, Almoutaz, Eltayeb, Mohamed Esmat Mohamed, Esraa, Essam, Mahmoud, Fahd, Osama, Farouk, Rabea, Gadelkareem, Ahmed, Ghoneim, Mohammed Hamada Takrouney, Ahmed, Hamdan, Abd El-Rahman Hamed, Ahmad, Hasan, Mohamed Abdelghafor Hassanin, Islam, Hawal, Kerollos, Henes, Mohamed Omar Herdan, Helal, F Hetta, Omar, Ibrahem, Mostafa, Ibrahim, Islam, H Ibrahim, Mahmoud, Kamel, Mohamed, Khallaf, Shrouk, M Elghazaly, Abobakr, Mahfouz, Osman, Mahmoud, Wesam, Mahran, Doha, Mahrous, Abadeer, Marsis, Ayman, Mohamed, Aliae Mohamed Hussein, Amr Mohamed Sayed, Mahmoud, M Mohammed, Ahmed, Mokhtar, Fatma, A Monib, Ahmed, Nageeb, Mariam Albatoul Nageh, Mohammed, Nageh, Nehal Gamal Omar, Abdelrahman, Ragab, Abdelrahman, Ramadan, Abdallah Rashad Temerik, Aya, Sabry, Hadeer, Safwat, Omar, Salah, Ahmed Elhussiny Salah Mahmoud, Ahmed, Saleh, Mahmoud, Sallam, Ahmed, Samir, Reem, Sayad, Esraa, Sayed, Ahmed, S Sedik, Mohammed, Shahine, Antonios, Soliman, Wael, Soliman, Mohamed Gamal Taher, Randa, Wanees, Ebrahim Ahmed Yousof, Ahmed, Youssef, Omar Zein Elabedeen, Ahmed Emad Sayed Hassan, Shrouk Abdel Fattah, Ahmed, Abostate, Samar, Ali, Saad Ali Saad Salama, Mohamed, Allam, Ahmed, Alsadek, Ebrahim, Arafa, Ahmed, Barakat, Mohammed El Sherpiny, Gehad, Elbehairy, Mahmoud, Eleisawy, 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Bahi, Abdurhman, Atea, Ibrahim, Gamal, Mohamed El Kassas, Wael, Omar, Ahmed, Tawheed, Saeid, Al-Oribi, Nuran Khaled Aly, Mahmoud, Elfiky, Alaa Eldine Elmaghraby, Abdulrahman, Elrahmany, Motaz, Elsherbeeny, Youssef, Helmy, Ahmed, Nabil, Ahmed Samir Farahat, Mostafa, Soliman, Amr, Wassef, Abdelrahman, Abdelrahman, Seif, Elsaban, Galal, Ghaly, Rana, Hamdy, Hamada, Mondy, Lubna, Abdallah, Abdelrhman Kz Darwish, Mohamed, Rabea, Mohammed, A Azab, Alzhraa Salah Abbas, Marwa, El-Deeb, Mohamed, Fawzy, Maher, Ibraheem, Abdelrahman, Bakry, Sarah, Elnems, Randa, Elsheikh, Mohamed Ibrahim Gbreel, Mahmoud, Hafez, Mohamed, Jammal, Khaled, M Hamam, Abdelrahman, M Makram, Omar Mohamed Makram, Salma, Rabie, Sherine Yousery Askalany, Mokhtar Mohamed Ibrahim Abushanab, Abdulrahman, Eid, Mohammed Refaat Ibrahiem Amin El Ghalid, Ahmed Adel Abdelaty, Elsayed, A Fayad, Asmaa, Radwan, Moataz, Sallam, Ahmad, Shokry, Ahmed Mohamed Farouk, Ahmed, Elfallal, Hossam, Elfeki, Mohamed, Elsaeed, Mahmoud, Elsaid, Amany, 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Abanga Alatiiga, Munira, Amadu, Nathaniel, Annan, German Azahares Leal, Mohammed, Bukari, Alexis, Buunaaim, Ernest, Cheyuo, Latif Daboo Salifu, Michael, Damah, Malcolm, Dery, Edem Kojo Dzantor, Odoniel Guerra Garcia, Yabasin Iddrisu Baba, Adamu, Issaka, Abdul-Jalilu Mohammed Muntaka, James, Murphy, Yaa Nyarko Agyeman, Wisdom Opoku Amankwaa, Imoro, Osman, Samuel, Pie, Anwar Sadat Seidu, Mohammed, Sheriff, Ana Maria Simono Charadan, Abraham, Titigah, Mundashiru, Yahaya, Musah, Yakubu, Edwin Mwintiereh Ta-Ang Yenli, Ioannis, Grypiotis, Nikolaos, Kiriakopoulos, Georgios, Koliopoulos, Vasilis, Kyvelos, Georgia, Micha, Triada, Papadopoulou, Dimitrios, Balalis, Evangelos, Fradelos, Dimitrios, Korkolis, Nicholas, Alexakis, Kyveli, Angelou, Dimitrios, Haidopoulos, Anastasia, Prodromidou, Alexandros, Rodolakis, Nikolaos, Thomakos, Dimitris, Psychogios, Pantelis, Antonakis, Konstantinos, Bramis, Leonidas, Chardalias, Ioannis, Contis, Nikos, Dafnios, Dionysios, Dellaportas, Papalouka, Dimitra, 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Santhosh Kumar Sampengere Annayappa, Tulika, Agrawal, Premkumar, Anandan, Aditya, Baindur, Manjunath, Bd, Savitha, C, S Anitha, G, Sandeep, Harigond, K Tejeswini, K, Venkatesh, Kesarla, Mallikarjuna, Manangi, B Hareesh, P, Jyothi K, R Ranjan, Athish, Shetty, Santhosh Shivashankar Chikkanayakanahalli, Tanvi, Sunil, Shreya, Syamala, Kavya, Tharanath, Aditya, Atal, Niranjana, Rajagopal, S Srinath, B, Vinod, Nk, L Thirumanikandan, P, Nitu, Mishra, Sushruta, Shrivastava, Rekha, Wadhwani, Reyaz, Ahmad, Zainab Ahmad Haq, Prateek, Behera, Ritika, Dhurwe, Rehan, Haq, Vaibhav, Jain, Anuj, Jain, Sunaina, Karna, Kameshwarachari, Pushpalatha, Sumit, Raj, John Ashutosh Santoshi, Pooja, Singh, Virendra, Verma, Vaishali, Waindeskar, Moorat Singh Yadav, Zaheda, Aziz, Koyel, Chakraborty, Preetam, Chappity, Gurudip, Das, Debajyoti, Datta, Saubhagya Kumar Jena, Madhabananda, Kar, Susanta, Khuntia, Pankaj, Kumar, Ravi, Kumar, Aswathi, Kv, Abhijeet, Mishra, Swastik, Mishra, Yash, Mittal, Dillip, Muduly, 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Reddy Voppuru, Anushri, Soni, Gira, Soni, Junaid, Alam, Dinesh, Bagaria, Minu, Bajpai, Akshay Kumar Bisoi, Arvind, Chaturvedi, Sandeep, Chauhan, Narendra, Choudhary, Rajendra Singh Chouhan, Sunil, Chumber, Kamran, Farooque, Vasubabu, Gudala, Amit, Gupta, Mohit, Joshi, Apoorva, Kabra, Shashank Sharad Kale, Abhinav, Kumar, Subodh, Kumar, Dhruv, Mahajan, Rajesh, Malhotra, P Ramesh Menon, Biplap, Misra, Samarth, Mittal, Rajinder, Parshad, Pratyusha, Priyadarshini, Sushma, Sagar, Pradeep Brijkishor Sharma, Vijay, Sharma, Vivek, Trikha, Mayank, Tyagi, Kuldeep, Bansal, Kalyan, Varma, Anurag, Mishra, Rajdeep, Singh, Rohit, Bhardwaj, Abhishek, Mittal, Sabarirajan, Ponnusamy, Gyan Ranjan Singh, Isha, Tuli, Pramod, Bhor, Sanjay, Dhar, Sachin, Kale, Manisha, Aggarwal, Himani, Gupta, Gurleen, Kaur, Ajinkya, Deshpande, Anup, Gadekar, Tanmay, Jaysingani, Taufiq, Panjwani, Rakesh, Patil, Narendra, Vaidya, Sunil Kumar Vishwakarma, Utkarsha, Wayal, Debnarayan, Dutta, Rajnish, Arora, Mohit, Dhingra, 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Venkateshwaran, Narasiman, Meer, Chisthi, Gejoe, George, Harihara, Jothi, Dilber, Pareed, Godwin David, C Mathew, Livingston, Abel, Mansi, Agrawal, Rabindranath, B, Manish, Baldia, Ravi Kishore Barla, Manisha, Beck, Santosh, Benjamin, Jeremy, Bliss, Lisa, Cherian, Sreekar, Devarakonda, Geley, Ete, Arun Jacob Philip George, Amish, Gohil, Deeptiman, James, Mark Ranjan Jesudason, Lallu, Joseph, Treasa, Joseph, Kathir, Joyson, Gomathi, Karnan, Albert, Kota, Pushplata, Kumari, Anitha, Loganathan, Vasanth Mark Samuel, John, Mathew, Senthil, K Nathan, Joby Elizabeth Ninan, Ajay, Philip, R Priyadarshini Priyadarshini, Suganya, S, Habie, Samuel, Gilbert, Samuel, Daniel, Selvaraj, Srujan, Sharma, Suraj, Surendran, D Hariharan, T, Santhosh Kumar Thangaraj, Vinotha, Thomas, Varghese, Thomas, John, K Thomas, S Harish, Y, Manobhiram, Boggavarapu, Murali Krishna Voonna, Marilaeta Cindryani Lolobali, Christopher, Ryalino, Tjokorda Gde Agung Senapathi, Mahadewa, Tjokorda, I Made Gede Widnyana, Dita, 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Massimiliano, Gennaro, Lorenzo, Giannini, Marcello, Guaglio, Marco, Guzzo, Andrea, Leva, Alberto, Macchi, Elena, Manzo, Fabio, Martinelli, Ilaria, Mattavelli, Vincenzo Maria Mazzaferro, Francesco, Raspagliesi, Luigi, Rolli, Laura, Sala, Roberto, Salvioni, Mario, Santinami, Silvia, Segattini, Luca, Sorrentino, Carlotta, Zaborra, Alexandre, Anesi, Gianmaria Casoni Pattacini, Mattia Di Bartolomeo, Francesca, Pecchini, Arrigo, Pellacani, Micaela, Piccoli, Federico, Fusini, Andrea, Gattolin, Marco, Migliore, Roberto, Rimonda, Diego, Sasia, Elisabetta, Travaglio, Federica, Brunetti, Marco, Cereda, Marco, Ceresoli, Luca, Cigagna, Cristina, Dell'Oro, Alessandro, Fogliati, Robert, Fruscio, Tommaso, Grassi, Maini Marzia Isabella, Luca Carlo Nespoli, Massimo, Oldani, Sara, Ornaghi, Nicolò, Tamini, Carmine, Antropoli, Antonio, Castaldi, Alessio, Palumbo, Gaia, Altieri, Umberto, Bracale, Francesco, Corcione, Marcello De Luca, Giovanni Domenico De Palma, Maria Michela Di Nuzzo, Ruggero, Lionetti, Dalila Loredana Lo Bue, Gaetano, Luglio, Gianluca, Pagano, Roberto, Peltrini, Nello, Pirozzi, Francesca Paola Tropeano, Alessia, Aversano, Andrea, Belli, Maria, D'Amico, Paolo, Delrio, Francesco, Izzo, Renato, Patrone, Daniela, Rega, Pietro, Maida, Ester, Marra, Gianpaolo, Marte, Andrea, Tufo, Francesco, Bianco, Antonio, Cappiello, Simona, Gili, Paola, Incollingo, Alessandra, Novi, Giulia, Bagaglini, Claudio, Iovino, Francesco Menegon Tasselli, Maria Paola Menna, Francesco Maria Romano, Settimio, Rossi, Guido, Sciaudone, Francesco, Selvaggi, Lucio, Selvaggi, Francesca, Simonelli, Guido, Coretti, Mario, Pannullo, Adolfo, Renzi, Francesca, Ascari, Giuliano, Barugola, Giacomo, Ruffo, Paolo, Baroffio, Paolo, Bellora, Laura Enrica Benedetti, Cristina, Cerri, Giordana, D'Aloisio, Maurizio, Ferrari, Elisa, Francone, Sergio, Gentilli, Herald, Nikaj, Antonella, Chessa, Alessandro, Fiorini, Luca, Campagnaro, Franco, Chioffi, Pietro, Ciccarino, Roberto, Colasanti, Francesco de Falco, Fotios, Kalfas, Gioacchino, Mattisi, Giulia, Nezi, Matteo, Palma, Angelica, Rizzoli, Domenico, Rossi, Davide, Russo, Renato, Salvador, Francesco, Volpin, Guido, Bissolotti, Stefano, Fusetti, Francesco, Lemma, Vito, Chiantera, Mariano Catello Di Donna, Giulio, Sozzi, Emanuele, Cammarata, Sofia, Campanella, Daniela, Canzonieri, Adriana, Cordova, Federico De Michele, Ettore, Dinoto, Mara, Franza, Leo, Licari, Daniele, Matta, Domenico, Mirabella, Felice, Pecoraro, Roberto, Pirrello, Pierfrancesco, Pugliese, Fernando, Rosatti, Giuseppe, Salamone, Francesca, Toia, Massimiliano, Tripoli, Cosimo, Callari, Dario Di Miceli, Alfredo, Annicchiarico, Luca, Bellanti, Michela, Bergonzani, Roberto, Berretta, Elisa, Cabrini, Vito Andrea Capozzi, Fausto, Catena, Federico, Cozzani, Paolo Del Rio, Marco, Domenichini, Anna, Fornasari, Antonio, Freyrie, Tiziana, Frusca, Mario, Giuffrida, Gennaro, Perrone, Giulia, Rossi, Matteo, Rossini, Andrea, Varazzani, Vittorio, Arici, Antonio, Bozzani, Lorenzo, Cobianchi, Matteo, Filardo, Marika Sharmayne Milani, Franco, Ragni, Andrea de Manzoni Garberini, Edoardo, Baldini, Diana, Carpaneto, Michele, Cauteruccio, Corrado, Ciatti, Luigi, Conti, Serena, Gattoni, Pietro, Maniscalco, Gerardo, Palmieri, Calogero Puma Pagliarello, Giuseppe, Caristo, Raffaele, Galleano, Michele, Malerba, Marcello, Calabrò, Francesca, Farnesi, Elia Giuseppe Lunghi, Andrea, Muratore, Nicoletta Sveva Pipitone Federico, Joel Reuben Abel, Lorenzo, Andreani, Darienzo, Antonio, Vittorio, Aprile, Riccardo, Balestri, Giacomo, Benettini, Stefano, Berrettini, Luca, Bruschini, Massimo, Chiarugi, Federico, Coccolini, Simone, Colangeli, Camilla, Cremonini, Lodovica Cristofani Mencacci, Iacopo, Dallan, Silvia De Santi, Gregorio Di Franco, Lorena Di Girolami, Giacomo, Fiacchini, Niccolò, Furbetta, Stylianos, Korasidis, Marco, Lucchi, Andrea, Morandi, Luca, Morelli, Serena, Musetti, Carlo Maria Neri, Matteo, Palmeri, Miriana, Picariello, Francesco, Porcelli, Marco, Puccini, Nicolo, Roffi, Erica, Statuti, Dario, Tartaglia, Alberto, Tonelli, Matteo, Vianini, Gianluca, Baronio, Mauro, Montuori, Enrico, Pinotti, Stefano Maria Massimiliano Basso, Federica, Maffeis, Paolo, Ubiali, Lorenzo, Aguzzoli, Saverio, Coiro, Giuseppe, Falco, Vincenzo Dario Mandato, Valentina, Mastrofilippo, Simone, Mele, Caterina, Baldi, Carlo, Corbellini, Gianluca Matteo Sampietro, Massimo, Dugo, Mauro, Garino, Chiara, Marafante, Antonio, Masciandaro, Elisabetta, Moggia, Alessandra, Murgese, Felice Eugenio Agro, Gabriella Teresa Capolupo, Filippo, Carannante, Marco, Caricato, Vincenzo, Denaro, Erica, Mazzotta, Rocco, Papalia, Giuseppe, Pascarella, Alessandro, Strumia, Biagio, Zampogna, Matteo, Cinquepalmi, Marco, Colasanti, Celeste Del Basso, Federica, Falaschi, Nicola, Guglielmo, Roberto Luca Meniconi, Alessandra, Pecoraro, Sofia, Usai, Giacomo, Crescentini, Antonella, Larcinese, Emanuele, Picone, Giovanni, Sinibaldi, Annamaria, Agnes, Salvatore, Agnes, Sergio, Alfieri, Francesco, Belia, Valentina, Bianchi, Giuseppe, Bianco, Alberto, Biondi, Paola, Campennì, Valerio, Cozza, Sabatino, D'Archi, Domenico, D'Ugo, Veronica De Simone, Marta Di Grezia, Sofia Di Lorenzo, Federica, Ferracci, Valeria, Fico, Gianluca, Franceschini, Pietro, Fransvea, Giulio, Gasparini, Luca, Gordini, Antonio La Greca, Francesco, Litta, Celestino Pio Lombardi, Angelo Alessandro Marra, Angelo, Parello, Marco Maria Pascale, Romeo, Patini, Gilda, Pepe, Roberto, Persiani, Caterina, Puccioni, Carlo, Ratto, Fausto, Rosa, Gianmarco, Saponaro, Lorenzo, Scardina, Tedesco, Silvia, Giuseppe, Tropeano, Maria, Benevolo, Daniele, Bugada, Flaminia, Campo, Maria Gabriella Dona, Valentina, Manciocco, Paolo, Marchesi, Riccardo, Mastroianni, Francesco, Mazzola, Silvia, Moretto, Raul, Pellini, Gerardo, Petruzzi, Barbara, Pichi, Giuseppe, Simone, Gabriele, Tuderti, Jacopo, Zocchi, Luigi Marino Cosentino, Andrea, Sagnotta, Roberta, Angelico, Vittoria, Bellato, Michela, Campanelli, Marzia, Franceschilli, Michele, Grande, Giorgio, Lisi, Tommaso Maria Manzia, Lorenzo, Petagna, Bruno, Sensi, Giuseppe, Sica, Leandro, Siragusa, Giuseppe, Tisone, Marco, Assenza, Barbara, Binda, Massimo, Biondi, Gioia, Brachini, Placido, Bruzzaniti, Mauro, Casagrande, Flavia, Ciccarone, Pierfranco Maria Cicerchia, Bruno, Cirillo, Daniele, Crocetti, Giancarlo, D'Ambrosio, Vito, D'Andrea, Francesca De Felice, Giorgio De Toma, Carlo Della Rocca, Giulia, Duranti, Pietro, Familiari, Enrico, Fiori, Giovanni Battista Fonsi, Alessandro, Frati, Stefania La Rocca, Filippo La Torre, Pierfrancesco, Lapolla, Giovanni, Marruzzo, Simona, Meneghini, Andrea, Mingoli, Andrea, Picchetto, Antonella, Polimeni, Diego, Ribuffo, Maurizio, Salvati, Antonio, Santoro, Paolo, Sapienza, Luigi, Simonelli, Valentino, Valentini, Martina, Zambon, Giuseppa, Zancana, Emma, Zuppi, Simone, D'Annunzio, Cosimo De Nunzio, Silvia, Fiorelli, Mohsen, Ibrahim, Chiara, Loffredo, Domenico, Massullo, Cecilia, Menna, Rocco, Monica, Massimiliano, Pelli, Erino Angelo Rendina, Leonardo, Teodonio, Andrea, Tubaro, Giulio, Argenio, Giulio, Accarino, Accarino, Giancarlo, Antonio Nicola Giordano, Luca, Cardinali, Elisa, Sebastiani, Grazia, Travaglini, Erika, Andreatta, Emanuele Luigi Giuseppe Asti, Daniele, Bernardi, Luigi, Bonavina, Caterina, Froiio, Andrea, Lovece, Chiara, Copelli, Alfonso, Manfuso, Pasquale Di Maio, Marco, Giudice, Oreste, Iocca, Rosario, Cennamo, Tommaso, Cornali, Francesco Di Marzo, Cristian, Altana, Francesco, Bussu, Giampiero, Capobianco, Anna Giacomina Carta, Sandro, Ciccarello, Maria Laura Cossu, Pietrina, Cottu, Giacomo De Riu, Salvatore, Dessole, Francesco, Dessole, Salvatora, Dettori, Carlo, Doria, Alessandro, Fancellu, Claudio, F Feo, Giorgio Carlo Ginesu, Giuliana, Giuliani, Marco Giuseppe Iannuccelli, Massimo, Madonia, Roberto, Mancino, Olindo, Massarelli, Gianfranco, Meloni, Fabio, Milia, Andrea, Mulliri, Teresa, Perra, Marco, Petrillo, Antonio, Piras, Franco, Piredda, Francesco, Pisanu, Alberto, Porcu, Davide, Rizzo, Angelino, Sanna, Antonio Mario Scanu, Fabrizio, Scognamillo, Damiano, Soma, Anna Rita Tanca, Alessandro, Tedde, Matteo, Tedde, Luigi Angelo Vaira, Andrea Bartalini Cinughi de Pazzi, Osvaldo Carpineto Samorani, Daniele, Fusario, Luigi, Marano, Gaia, Oldrà, Anna Lisa Pesce, Stefania Angela Piccioni, Luca, Resca, Vincenzo, Ricchiuti, Franco, Roviello, Vinno, Savelli, Alberto, Abrate, Pierpaolo, Bordoni, Lorenzo, Bosio, Guglielmo, Clarizia, Francesco, Fleres, Marco, Franzini, Antonio, Fratto, Pierluigi, Giumelli, Alessandro, Grechi, Alessandro, Longhini, Fabrizio, Lorusso, Elisa, Scarnecchia, Federica, Scolari, Alessandro, Spolini, Vincenzo, Maiuri, Matteo, Papandrea, Arturo, Roncone, Lorenzo, Conti, Andrea, Rizzi, Marco, Rovagnati, Alberto, Brolese, Tommaso, Cai, Francesco Antonio Ciarleglio, Francesca, Dalprà, Gianni, Malossini, Liliana, Mereu, Irene, Tamanini, Saverio, Tateo, Giovanni, Viel, Enrico, Battistella, Paolo Boscolo Rizzo, Cristoforo, Fabbris, Marco, Massani, Giacomo, Spinato, Roberta, Tutino, Ugo, Grossi, Alessandro, Iacomino, Simone, Novello, Maurizio, Romano, Serena, Rossi, Giulio, Santoro, Giacomo, Zanus, Sokol, Trungu, Giada, Aizza, Gabriele, Bellio, Selene, Bogoni, Marina, Bortul, Biagio, Casagranda, Sara, Cortinovis, Nicolò de Manzini, Davide, Drigo, Paola, Germani, Manuela, Mastronardi, Lucia, Paiano, Silvia, Palmisano, Pier Luigi Filosso, Francesco, Guerrera, Matteo, Marro, Mauro, Rinaldi, Enrico, Ruffini, Stefano, Salizzoni, Laura, Bardelli, Mattia, Berselli, Giacomo, Borroni, Eugenio, Cocozza, Matteo, Desio, Bottazzoli, Elisa, Giuseppe, Ietto, Valentina, Iori, Domenico, Iovino, Lorenzo, Livraghi, Valentina, Marchionini, Stefano, Megna, Emma Amal Nahal, Mara, Palumbo, Valeria, Quintodei, Alessandra, Zullo, Lucrezia, D'Alimonte, Giovanni, Pirozzolo, Chiara, Vignotto, Tommaso, Campagnaro, Andrea, Caravati, Simone, Conci, Carlotta De Cristofaro, Gabriele, Gecchele, Tommaso, Giuliani, Jacopo, Graziosi, Alfredo, Guglielmi, Salvatore, Paiella, Corrado, Pedrazzani, Tommaso, Pollini, Simone, Rattizzato, Andrea, Ruzzenente, Roberto, Salvia, Giulia, Turri, Matilde, Bacchion, Giovanni, Butturini, Andrea, Casaril, Alessandro, Giardino, Harmony, Impellizzeri, Marco, Inama, Frigerio, Isabella, Gianluigi, Moretto, Marco De Zuanni, Enrica, Deiana, Mario, Guglielmo, Alessandro, Broglia, Claudia, Casarini, Caterina Costanza Zingaretti, Marta, Bonaldi, Giovanni, Cesana, Francesco, Mastriale, Stefano, Olmi, Matteo Uccelli Yasuyuki Fukami, Takuya, Saito, Tsuyoshi, Sano, Naoki, Hirai, Kazuyoshi, Hirota, Tetsuya, Kushikata, Tasuku, Oyama, Junichi, Saito, Katsuhiko, Ishibashi, Mizue, Kamiyama, Kyongsuk, Son, Kentaroh, Tarao, Takayuki, Yamada, Toshiya, Shiga, Chisaki, Aze, Yoshihiko, Deguchi, Hirotoshi, Hasegawa, Tatsuki, Hoshino, Yasushi, Innami, Hiroyuki, Inoue, Shingo, Ito, Takeshi, Nomura, Tomomi, Ogihara, Reina, Okada, Takashi, Ouchi, Yuri, Sekiya, Keikoku, Tachibana, Emi, Takano, Masae, Yamamoto, Shuko, Matsuda, Yuka, Matsuki, Kenji, Shigemi, 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Yu, Mitsuru, Ida, Masahiko, Kawaguchi, Kenji, Kawamura, Munehiro, Ogawa, Hiroshi, Okada, Shunji, Endo, Yoshinori, Fujiwara, Masaharu, Higashida, Hisako, Kubota, Toshimasa, Okada, Hironori, Tanaka, Tomio, Ueno, Kazuhiko, Yoshimatsu, Motohiro, Kikukawa, Akira, Kuriyama, Susumu, Matsushime, Hishikawa, Hidehiko, Haruaki, Hino, Yoji, Hisamatsu, Akio, Kamiya, Hidefumi, Kinoshita, Masato, Kita, Toshinori, Kobayashi, Taku, Michiura, Hirokazu, Miki, Tomohiro, Murakawa, Hidetaka, Okada, Tomohito, Saito, Ryoichi, Saito, Motohiko, Sugi, Genichiro, Sumi, Yamaki, So, Tomohisa, Yamamoto, Aya, Yoshida, Tatsuya, Kambara, Sayaka, Kanematsu, Okazaki, Satoshi, Takeshi, Hijikawa, Hiroaki, Kitade, Hidesuke, Yanagida, Tomoyuki, Fujita, Satsuki, Fukushima, Naoki, Tadokoro, Taku, Furukawa, Yusuke, Iizuka, Yuji, Otsuka, Masamitsu, Sanui, Ikumi, Sawada, Hideki, Iwahashi, Morikazu, Miyamoto, Masashi, Takano, Tsutomu, Mieda, Chihiro, Ando, Tetsuro, Isada, Taku, Ishizaki, Qaed Bani Amer, Sachi, Ishida, Yasuma, 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Rakotoarisoa, Maharo, Ramifehiarivo, Solonirina Harinarindra Ranaivoson, Hajamihamina Marina Parfaite Randriantsoa, Narindra Njarasoa Mihaja Razafimanjato, Jeannie Ba Razafindrahita, Solofoarimanana, Solofoarimanana, Guillaume Odilon Tsiambanizafy, Rakotonaivo Mamisoa Judicaël, Corinne Eulalie Solo, Tojomamy Herinjaka Ralaizafindraibe, Fanonjomahasoa Safiry Andofenohasina, Hanta, Rasataharifetra, Vanona Barijaona Razafindraibe, Angelin Sablon Herinirina, Andre, Das, Thanga, Ganapathy, Yohesuwary, Gunarasa, Kalyani, Mariapan, Maathichsudhaar, Muniandy, Mohd Firdauss Osman, Mohd Azem Fathi Mohammad Azmi, Jien Yen Soh, Wan Zainira Wan Zain, Andee Dzulkarnaen Zakaria, Zaidi, Zakaria, Kheng Hooi Chan, Firdaus, Hayati, Syamim, Johan, Jin Jiun Mah, Ratha Krishnan Sriram, Sentilnathan, Subramaniam, Nur Shahirah Binti Muhammad Shahimi, Khairul Hazim Hamdan, Mohd Razali Ibrahim, Guhan, Muthkumaran, Ju Ann Tan, Mohana Raj Thanapal, Dayang Anita Abdul Aziz, Mae-Lynn, Bastion, Mohd Hairul Nizam Harun, Farrah-Hani, Imran, Norshamsiah Md Din, Ayesha Mohd Zain, Mushawiahti, Mustapha, Syed, Nabil, Ainal Adlin Naffi, Abd Jabar Nazimi, Roszalina, Ramli, Bee Hong Soon, Pavin, Bal, Joo Qing Cheng, Mohd Rusdi Draman, Ahmad Nazran Fadzli, Sakina, Ghauth, Weipin, Hung, Suniza, Jamaris, Peng Soon Koh, Sivakumar, Krishnasamy, Yeong Sing Lee, Yew Toong Liew, Jeffery Zk Lim, Raymond Chung Siang Lim, Hiong Chin Lim, Siti Farhan Moh Pauzi, Shireen Anne Nah, Ashvin Krishna Nair, Doris Sin Wen Ng, Yuki Julius Ng We Yong, Kamarajan, Ramayah, Rahmah, Saaid, Anand, Sanmugam, Mee Hoong See, Kanesh Kumaran Seevalingam, Neha, Sethi, Srihari, Singaravel, Syeda Nureena Syed Jafer Hussain Zaidi, Chu Yik Tang, Li Ying Teoh, Kian Boon Wong, Ruben, Xavier, Ahmad Shuib Yahaya, Norhafiza, Ab, Rahman, Islah Munjih Ab, Rashid, Mohd Fahmi Abd Aziz, Mohd Norhisham Azmi Abdul Rahman, Nasser Muhammad Amjad, Shahidah Che Alhadi, Faisal, Elagili, Mohd Nazli Kamarulzaman, Azmi Md Nor, Ahmad Faidzal Othman, Mohd Yusof Sainal, Mat Salleh Sarif, Richelle, Chua, Aini Fahriza Ibrahim, Abdul Rauf Bin Ahmad, Yii Ling Lau, Ka Yee Liew, Kandasami, Palayan, Hui Yu Wong, Jasiah, Zakaria, Nor Azimah Abd Aziz, Norazila Abdul Rahim, Noorneza Abdul Rahman, Bahiyah, Abdullah, Mohamed Faizal Bin Sikkandar, Fatin, Firman, Nik Mohd Hazleigh, Intan Kartika Kamarudin, Shahril, Khalid, Mohd Fairudz Mohd Miswan, Akmal Zulayla Mohd Zahid, Norasyikin, Mustafa, Kadhim, Obaid, Mohd Yusoff Bin Yahaya Yahaya, Ahmad Ramzi Yusoff, Predrag, Andrejevic, Jeremy Borg Myatt, Svetlana, Brincat, Clifford, Caruana, Kurt Lee Chircop, Francesca, Chircop, Daniel Mario Chircop, Charles, Cini, Viktoria, Czok, Melanie, Farrugia, Gabriella, Grima, Fatimah Mj Hassan Almukhariq, Esther, Muscat, Darryl, Pisani, Mohsin Hassan Khan Roshan, Nicholas, Schembri, Mohamed, Shoukry, Neville, Spiteri, Gilbert, Tanti, Ziad Aboharp Hasan, Alberto Bazan Soto, Cedillo Rodrìguez Jonathan Rubén, Gustavo Esteban Lugo Zamudio, David Gerardo Miranda Gómez, José Alfredo Pérez Meave, Victor Pinto Angulo, Antonio Rojas Aguilar, Erik Efrain Sosa Duran, Alejandro Vàzquez Perez, Diego, Arroyo, Jose, Corona-Cruz, Hector, Martinez-Said, Heriberto, Medina-Franco, Manuel, Mendez, Teresa Moreno, Y Suárez Ma, Dolores Orta Díaz, Martha Patricia Pérez de León Vázquez, Angélica Azucena Soto Carvajal, Fidel Soto Hernández, Juan Ignacio Stenner, Adriana Valenzuela López, Mauricio Rene Hernandez, Rey Jesus Romero, Aldo Bernal Hernandez, Ana Olivia Cortes-Flores, Carlos Jose Zuloaga Fernandez Del Valle, Carlos Andres Colunga Tinajero, Jaime, Orozco-Perez, Laura Gabriela Peña Balboa, Guillermo, Yanowsky-Reyes, Víctor Hugo Alcalá Torres, Francisco José Barbosa Camacho, Irma Valeria Brancaccio Perez, Miguel Angel Calderon-Llamas, Aldo Camacho Gomez, Edgar-Joaquin, Cortes-Torres, Esteban, Cueva-Martinez, Oscar Durán Anguiano, Isaac Esparza Estrada, Roberto, Fierro-Rizo, Paola Flores Becerril, Clotilde Fuentes Orozco, Miguel García Castillo, Alejandro Gonzalez Ojeda, Ana Sofia Gonzalez Rubio, Erick Alonso González-García de Rojas, Luis Humberto Govea-Camacho, Jose Aldo Guzman Barba, Juan Carlos Ibarrola Peña, César Íñiguez Martínez, Francisco Javier Llamas-Macias, Ruben Eduardo Morán Galaviz, Cesar, Nuño-Escobar, Oscar-Everardo, Olvera-Flores, Jaqueline, Osuna-Rubio, Jose Victor Pérez-Navarro, Rodrigo, Prieto-Aldape, Luis Ricardo Ramirez Gonzalez, Marco Vinicio Ramírez Sánchez, Emilio, Reyes, Olaya Moramay Romero-Limón, Carlos Beas Ruiz-Velasco, José Antonio Sánchez Martínez, Guillermo, Sanchez-Villaseñor, Ismael, Sedano-Portillo, Gabriela Ambriz Gonzalez, Pedro Berrones Moreno Berrones Moreno, Ana Beatriz Calderon Alvarado, José de Jesús Cárdenas Barón, Araceli, Hernández, Francisco Javier León Frutos, Edgard Efren Lozada Hernandez, Eduardo Morales Valencia, Karla Daniela Pérez, Francina, Bolanos-Morales, Julio, Herrera-Zamora, Patricio, Santillan-Doherty, David Omar Arriaga Zavala, Salvador Francisco Campos Campos, Lizette, Carmona, Antonio Galindo Nava, Blanca, Santos, Juan Roberto Torres Cisneros, Katya, Bozada-Gutiérrez, Jose Jesus Herrera, Mucio Moreno Portillo, Francisco Pérez López, Mario, Trejo-Avila, Carlos Valenzuela Salazar, Maria Regina Alvarez, Fernando, Cordera, Marco Antonio de la Rosa Abaroa, Antonio, Gómez-Pedraza, Roberto, Hernandez, Antonio, Maffuz-Aziz, José Antonio Posada, Carlos Robles Vidal, Jose-Luis, Beristain-Hernandez, Blas Eduardo Quintero Sada, Nicolás Uxmal Soruco Heredia, Ernesto, Anaya, Ricardo, Arceo-Olaiz, Bernardo Gutiérrez Sougarret, Sonia Lopez Flores, Francisco, J Ochoa Carrillo, Raymundo Alfredo Pérez Uribe, Ludwigvan Adriano Bustamante Silva, Arianne, I Lupián-Angulo, Christian Enrique Soulé Martínez, Francisco César Becerra García, Ernesto Javier Guerrero Casillas, Edgar, Uribe, Nereida Esparza Arias, J Mindy Hernández-Nava, David, Isla-Ortiz, Javier, Melchor-Ruan, Esmeralda Romero Bañuelos, Rosa, Salcedo-Hernández, Diana, Vilar-Compte, Paola Zinser Peniche, Jorge, L Aguilar Frasco, Alejandro, Alfaro-Goldaracena, Gabriela Alejandra Buerba, Emma, Castro, Carlos, Chan, Ruben, Cortes, Ismael, Dominguez-Rosado, Daniel Garay Lechuga, Miguel, Herrera, Carlos, Hinojosa, Gabriel, Lopez-Pena, Edgar, Martos, Miguel Ángel Mercado, Paulina Moctezuma Velázquez, Roberto, Moguel, Mariano, Oropeza, Emmanuel Peña Gómez Portugal, Oscar Emmanuel Posadas-Trujillo, Noel, Salgado-Nesme, Oscar Santes Jasso, Catherine, Sarre, Karla Yareli Hernandez Skewes, Cristian, Jimenez, Roque Delfino Licona-Meníndez, Guillermo, Montiel, Alberto, Navarrete-Peón, Shannat, Ortega, Fernanda Romero Lechuga, Oscar, Roque, Fernando, Sevilla, Rogelio, González-López, Roberto Ángel Núñez-González, José Antonio Ortega-Jiménez, Antonio, Alvarado, Alfonso Alvarez Manilla Orendain, José Alberto Atristain Pesquera, Diego, Frutos, Felipe, Garcia, Juan Antonio González León, Marlin, Guzman, Dorihela, Herappe, Jesús Giovanni Inzunza Miranda, Miguel Angel Jimenez Botello, Ricardo, Lerma, Fabián Hilario Mendoza Pedraza, Monica Noguez Castillo, Jose Anatolio Resendiz, Oscar Alejandro Sánchez García, Laura Elena Fernandez Rios, Pamela, Frigerio, Iset, Garcia, Gloria Isela Mendoza Frías, Emmanuel, Moreno, Raquel, Quiroga, Alejandra Anzures Mendoza, Arantza Govela Hinojosa, Aldo, Izaguirre, Luis Hdez Miguelena, Jose Manuel Luna Vazquez, Indalecio Barcelata Rodriguez, Raul Ramos Mange, Carlos Reyes Utrera, David Zarate Saenz, Miguel Angel Zavala Gonzalez, Francisco Andrade González, Ludivina, Cortes, David, Roman, Aaron, Sanchez, Radu, Gurghiș, Gheorghe, Rojnoveanu, Marin, Vozian, Petru, Caraja, Gheorghe, Croitor, Roman, Mosneaga, Sarnai, Erdene, Lundeg, Ganbold, Sergelen, Orgoi, Soyombo, Orsoo, Altanchimeg, Sainbayar, Erdene, Sandag, Burmaa, Sanjaa, Rita Ait Benhamou, Abdessamad El Azhari, Bennis, Ghita, Sara, Lhassani, Sidi Mamoun Louraoui, Mounir, Rghioui, Sarra, Saaf, Taha, Gadouali, Iltimass, Gouazar, Moniba, Korch, Ahmed, Letrache, Amina, Louari, Anas, Zamame, Samia, Ousouss, Meryem, 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Alexander, Borgstein, Suzanne, Gisbertz, Sarah, Mikdad, Mark van Berge Henegouwen, Roel, Bakx, Marc, Besselink, Daniel Hl Lemmers, Anne-Jasmin, Roelofs, Paul van Amstel, Claire van Helsdingen, Ludi, Smeele, Niels, Harlaar, Frederik, Jonker, Sjirk van der Burg, Justin, Y van Oostendorp, Saranda, Ombashi, Tim van der Voort, Martijn van Geldorp, Lisanne, Posma-Bouman, Hans Donald de Boer, Annette, Olieman, Henriette, Smid-Nanninga, Jean-Paul P, M De Vries, Rianne, Hogenbirk, Tyche, Derksen, Josephine, Franken, Steven, Oosterling, Peter, Nolte, Jelle van der List, Ian, Alwayn, Okker, Bijlstra, Andries, Braat, Ruth, Bulder, Michèle de Kok, Robin, Faber, Ben, Goudsmit, Jaap, Hamming, Sven, Mieog, Alexander, Vahrmeijer, Fenna E, M van de Leemkolk, Joost van der Vorst, Jan van Schaik, Merel, Verhagen, Kim, Albers, Larsa, Gawria, Harry Van Goor, Michiel, Warle, Tessa, M van Ginhoven, Charlotte, Viëtor, Evert-Jan, Boerma, Lara, Lallitsch, Donald, Schweitzer, Wouter, Leclercq, Julie, Sijmons, 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Asensio, Luis Asensio Gomez, Diego, Carrion, Nuria, Chavarrias, Alvaro de Arriba, Jose Antonio Gazo Martínez Gazo, Alberto Gegúndez Simón, Juan Gómez Rivas, Natalia Gonzalez Alcolea, Carolina, Gonzalez-Gomez, Sara Gortázar de Las Casas, María Alexandra Heras Garceau, Alicia Hernández Gutierrez, Luis, Lassaletta, Fuad Samir Lopez Fernández, Marta, Mancheño, Blanca, Mateos-Serrano, Isabel Pascual Miguelañez, Maria Fernanda Pedrero Escalas, Julio, Peñarrocha, Yolanda, Perez, Maria Isabel Prieto-Nieto, Teresa Rivera Schmitz, Laura, Rodrigáñez, Ines, Rubio-Perez, Isabel Sanchez Cuadrado, José Ignacio Sánchez Méndez, Jorge Fernando Tone, Carlos Toribio Vazquez, Aitor, Urbieta, Diego Córdova García, Francisca Garcia-Moreno Nisa, Laura, Jiménez, Inmaculada, Lasa, Belen, Matías-García, Fernando, Mendoza-Moreno, Marina Pérez González, Ana Sánchez Gollarte, Alejandro Sanchez Pellejero, Cristina Vera Mansilla, Rafael, Barberá, Lourdes, Montes-Jovellar, Sara Fra Fernández, Nicolas Moreno Mata, Barbara, Algar-Yañez, Gutiérrez, Bérénice, Olga De La Varga-Martínez, Paula, Fernandez-Valdes-Bango, Sandra Maria Gadin-Lopez, José María Martínez-Gómiz, Elena, Nieto-Moreno, Ana, Nieto-Moreno, Montserrat Ocampos Hernandez, Maria Laura Pelegrina-Lopez, Alicia Ruiz Escobar, Elena, Sáez-Ruiz, Beatriz, Vazquez-Rivero, Antonio, Lara, Belén Vielva Del Campo, Fernando Alcaide Matas, Simbad, Costas-Ochoa, José María García Pérez, Elizabeth, Marmol, Paula Troncoso Pereira, Maria Del Pilar Concejo Cutoli, Juan Ramón Gómez López, Juan Carlos Martín Del Olmo, Paloma Guillamot Ruano, Javier Páramo Zunzunegui, Antonio Albarracín Marín Blázquez, Alfonso, Aliaga-Sanchez, Marta María Arroyo Domingo, Miriam Artés Artés, Ana Isabel Avellaneda Camarena, Aida, Blaya, Emny Rochell Bobadilla Romero, Pedro Vicente Fernández-Fernández, Lorena Galindo Iñiguez, Víctor Javier García Porcel, Clara Giménez Francés, Antonio Javier Gomez Poveda, Rebeca Gonzalez Celdran, Francisco Miguel González Valverde, Laura Guillamon Vivancos, Elena, Gurrea-Almela, Alejandra Jara Maquilón, Jose David Jimenez Parra, Abd Al Aziz Lanagrán Torres, Raquel Lax Perez, Carmen Maria Lopez Lopez, Francisco, Machado, Alfonso Marco Garrido, Florencio Manuel Marin Martinez, Victoria Martínez Muñoz, Esther, Medina, Olimpia, Molina, José Manuel Muñoz Camarena, Julian, Onate, Pedro Antonio Parra Baños, Emilio Peña Ros, Enrique, Rubio, Marina Sánchez Robles, Carlos Sanchez Rodriguez, María Valero Soriano, María Teresa Yepes García, Felipe, Alconchel, Andrés Balaguer Román, Valentín, Cayuela, Ana, Conesa, Ana Delegido García, Pedro José Gil Vázquez, Paula Gómez Valles, Francisco, Gómez-Bosch, Vicente, J León-Muñoz, Francisco, Martínez, Alvaro, Navarro-Barrios, Tatiana, Nicolás-López, Pablo Ramirez Romero, Emiliano, Cano-Trigueros, María Concepcion Alonso González, Leticia Gómez Viana, Ana Pastor Zapata, Elisa Contreras Saiz, Tamara Díaz Vico, Maria, Fernandez-Hevia, Luis Joaquín García Flórez, Raquel, Rodríguez-Uria, Sandra, Sanz, Lorena, Solar-Garcia, Aida Suárez Sánchez, Jaume, Gelonch, Dietmar, Reinaldo, Miriam Abdulkarim Polo, Asier, Aguirre, Marta Alomar Bofill, Santiago, Baena, Ashish, Bartakke, Carmen Cabeza Oliver, Marta Castro Suárez, José Andrés Cifuentes Rodenas, Olga Claramonte Bellmunt, Ant Onia Crespí Mir, Anabel De La Llave Serralvo, Mar, Escales, Isabel Rosa Fernández Burgos, Pablo Gandia Gonzalez, Juan Lliteras Jorge, María Cristina Martínez Canto, Javier, Mata, Andreu, Morell, Naila, Pagès, Sara Pérez Palao, Cristian Plaza Valiente, Catalina Ramon Barcelo, Carmen Reyero Fernández, Albert Reyes Claret, Leticia Rodriguez Vaquero, Maialen Saez de Vicuña Salinas, Pilar Santos Cidon, Concepcion, Segura, Aina, Serra, Maria Serra Guivernau, Maria Soler Pedrola, Marta, Tasende, Marta Torrent Lluch, María Inmaculada Valldeperas Hernández, Rosa-Maria, Yañez-Lpez, Andrea, Craus-Miguel, Laura Fernández Vega, Natalia Pujol Cano, Juan José Segura-Sampedro, Laura Colet Oliver, Ivan Dot Pascuet, Mireia, Duart, Isabel Tello Galindo, Eduardo Arrea Salto, Jaume Cámara Cabrera, Marta Camats Terré, Juan Gabriel Castro Ríos, Marta de la Rosa-Estadella, Anna, Fernandez-Colorado, Anna, Gasulla-Rodriguez, Antonio, Gimenez-Gaibar, Elena, Gonzalez, Yeray Maldonado Sotoca, Matilde, Molina-Corbacho, Montserrat Monfort Mira, Josep, Rodoreda, Marta Santos Espí, Arnau Verdaguer Figuerola, Elisa Angela Diego-Alonso, Jaime, López-Sánchez, Luis, Muñoz-Bellvis, Jacobo, Trebol, Borja, Aguinagalde, Cristina Gabriela Alzate Arsuaga, Ainhoa Andres Imaz, Iñigo, Arana, Coro Aranzabal Urrutia, Mikel, Armendariz, Lorena, Arrabal, Iñigo, Augusto, Adolfo, Beguiristain, Laura, Busto, Ainhoa, Cuevas, Itziar De Ariño, Alejandro, Elúa, Leticia, Fernandez, Arantza, Fernandez-Monge, Lander, Gallego, Alba, Garcia, Amaia Garcia Dominguez, Gregorio, Garmendia, Carmen, Grañén, Pelayo Hevia Rodríguez, Maria, Iraola, José Miguel Izquierdo, Aintzane, Lizarazu, Jon Ander Lizarbe, Claudia Cristina Lopes Moreira, Iker, López, Ruth Marquina González, Tania, Pastor, Adrián Recio Ayesa, Ana Paula Riverola Aso, Carballo, Rodríguez, Araceli, Rodrìguez, Maria Inmaculada Ruiz Montesinos, Patricia, Torres, Jose, Undabeitia, Ainara, Villafruela, Maria Barrionuevo Ramos, David, Blanco, Marta, Calderón, Eva Corral Rubio, Angel, Cuadrado-García, Daniela, Cubek, Ignacio, Fariña, Maria Fernandez LLorente, Natalia Flores Amador, Mariana García Virosta, Luis Garcia-Sancho Tellez, Adelina Gonzalez Martinez, María, Hernandez, Rubén Herreros Ruiz-Valdepeñas, Carmen Jimenez Sanchez, Eduardo Llamazares Cobo, María, López, Javier, López-Martin, Teresa Martínez Marivela, Luis Miguel Martinez Parra, Paloma Maté Mate, Paloma, Medrano, Manuel Moriche Carretero, Sara Núñez O'Sullivan, Manuel Ortega Oria de Rueda, María Begoña Pastor Nieto, Clara, Pérez, Antonio, L Picardo, Remedios Revilla Amores, Carmen Rodríguez Haro, Jose Alberto Rojo López, Fatima Sanchez Cabezudo Noguera, Cristina, Sanz, Daniel Serralta de Colsa, Cristina Valor Garcia, Marta Velasco Martinez, Iván Andújar Lara, Aranzazu, Calero-Lillo, Enrique, López-Ruiz, Alberto Díaz García, Alejandro Menéndez Moreno, Luis Eduardo Pérez-Sánchez, Ángela, Pascual, Miguel Adeba García, Marta Alonso Fernández, Javier Alvarez Gama, Roberto, Ballestero, Loreto Berjon De La Vega, Paola Calleja Hermosa, Mariana, Carrillo-Rivas, Alfonso, Casado, Andrea, Cerveró, Bonifacio Cimadevilla Calvo, Patricia Corriols Noval, Paloma de la Dehesa Cueto-Felgueroso, Miren Jasone Diez Zapirain, José Estevez Tesouro, José Antonio Fernández-Dívar Sánchez, Lucia Garcia Alcalde, Juan García Cardo, Tito García Moreno, Daniel, García-López, Belen Garcia-Montesinos Perea, Miren Gonzalez Benito, Rocio, Gonzalez-Aguado, Miguel Ángel Gordo Vega Gordo-Vega, Jose, Gutierrez-Banos, Jaime Jimeno Fraile, Yolanda Jubete Castañeda, Patricia Lopez Gomez, Antonia Jesús López López, David Mato Mañas, Carmelo, Morales-Angulo, Dieter, Morales-Garcia, Sara, Naranjo, Jose Luis Rabago, Jose Manuel Rabanal, Rocio Revuelta Zorrilla, Adriana Reyes Echeverría, Juan Carlos Rodríguez-Sanjuán, Juan Ramón Sanz, Isabel Simal Badiola, Guillermo, Tejón, Enrique Toledo Martínez, Raquel Varea Malo, Begoña, Fadrique, Tania, Funes, Elena González Revilla, Isaias, Alarcón, Daniel Aparicio Sánchez, Pablo Beltran Miranda, Carmen, Cepeda-Franco, María Josefa Cuevas López, Sandra, Dios-Barbeito, Virginia María Durán Muñoz-Cruzado, Carlos Gonzalez De Pedro, Rosa, M Jimenez-Rodriguez, Francisco, Manresa-Manresa, Gabriel, Marin, Cristobalina, Martin-Garcia, Sara, Martínez-Núñez, Juan Manuel Martos Martinez, Lucas, Mengíbar, Felipe Pareja Ciuro, Eduardo Perea Del Pozo, Veronica Pino Diaz, José, Pintor-Tortolero, Claudia, Quintero-Pérez, Maria Luisa Reyes Diaz, Mercedes Rubio Manzanares Dorado, Alejandro Sanchez Arteaga, Ana, Senent-Boza, Luis, Tallon-Aguilar, Jose, Tinoco-Gonzalez, Luis-Cristobal, Capitan-Morales, Juan Cintas Catena, Javier, Valdes-Hernandez, María Del Campo Lavilla, Alina López De Fernández, Miriam Abellan Lucas, Alba Diaz Padillo, Maria Gómez Romero, Linda Grace Puerto Tamayo, El Mostafa El Yaqine Er Raoudi, Melody García Domínguez, María Ángeles Gascón Domínguez, Marta González Pérez, Alba Hernáez Arzoz, Joana San Anton, Cristian, Ruminot, Hander Guillermo Acosta Diaz, Daniella, Laguado, Kristina, Aghababyan, Alba, Jr, Enrique, Artigues, Rocío, Belda, Miguel, Beltran, Jose Ignacio Blanes, Elena Campos Carot, Miriam, Cantos, Laura, Costa, Carolina Ferrer Gomez, José María Gallego Sánchez, Oscar, Gil-Albarova, Juan, Gilabert-Estellés, Emilio Lopez Alcina, Isabel López Sánchez, Severiano Marin Bertolin, Carolina, Martinez-Perez, Antonio Melero Abellán, Claudia Mulas Fernández, Beatriz, Novoa, Ruth, Nuñez, Pablo Renovell Ferrer, Guijarro-Jorge, Ricardo, Carolina Soledad Romero Garcia, Miguel Sanfeliu Giner, Silvia, M West, Enrique, Zapater, Ignacio, Miranda, Ana, Monís, Alejandro Roselló Añón, María José Sangüesa, Jorge Sanz Romera, Marcos, Adrianzén, Maria Barrios Carvajal, Mireia, Bauzá, Ana Benítez Riesco, Jose Antonio Carbonell Lopez, María-Carmen, Fernández-Moreno, Ricardo Gadea Mateo, Francisco, Garcia, Maris Luisa García-Pérez, Luisa Paola Garzon, Ana, Izquierdo, Maria Lapeña Rodríguez, Carlos, León-Espinoza, Fernando, Lopez, Rosa Martí Fernández, Sara Martínez Castro, Berta Monleón López, Isabel Mora Oliver, David, Moro-Valdezate, Ernesto Muñoz Sornosa, Sara Palomares Casasús, Leticia Pérez Santiago, Mara Albert Fort, Reyes, Balanzá, Lorena Bermell Marco, Carla Carratalá Pérez, Claudia Pilar Clemente Tomas, Marta, Córcoles, Jose Ángel Diez Ares, Carlos Domingo Del Pozo, Nuria Estellés Vidagany, Gonzalo, Garrigos, Mar Gimeno Gimeno Vicente, Blanca Alastrue Giner, Segundo, Gomez-Abril, Paloma, Gonzalez, Paula Gonzálvez Guardiola, Guillermo Sebastian Martínez Fernández, Charo Martínez García, Lucía, Martínez-Costa, Luz María Moratalla Charcos, Sergio Navarro Martínez, Carmen, Payá-Llorente, Álvaro Pérez Rubio, Nuria, Peris, Eduardo Picazo Pineda, Jordi Planelles Gómez, Francisco Ripoll Vidal, Andrea, Roca, Enrique, Salmerón-González, Raquel Segura Roselló, Ramon Trullenque Juan, Cristina Verdejo Gimeno, Jose Miguel Zaragozá García, Gema Zomeño Bravo, Marcos Bruna Esteban, Hanna, Cholewa, Milton de Jesus, Matteo, Frasson, Luis Oliver García, Héctor, J Aguado, Antonio Jose Alonso Villalba, Francisco, Ardura, Carmen Elena Badillo Bercebal, María Bedate Núnez, Alvarez-Ramos Begoña Aranzazu, Juan Beltrán de Heredia, Hugo, Bermejo, Juan Berrocal Cuadrado, Maria Bragado González, Juan, Bustamante-Munguira, Raul Calvo Gonzalez, Yolanda, Carrascal, Ana Belén Casas Marcos, María Carmen Cervera, Sergio Chávez Valladares, Ángel Cilleruelo Ramos, Susana, Cortes, Jesús, Crespo-Sanjuán, María, Cuaresma, Luis Antonio Cuellar Martin, Carlos Ferreras García, Alvaro, Fuentes-Martín, Abel, Ganso, Elena García García, Luis, Garcia-Florez, Virginia, Garcia-Virto, Juan, Gatón, Julio Alberto Gobernado Tejedor, Tania Gómez Sanz, Jose Ignacio González Martín, María Begoña Gregorio Crespo, Ana Herranz Arriero, Jose, Herreros, Marta Ibáñez Nieto, César, Infante, Carlos, Jezieniecki, Moises, Juarez, Gregorio de Jesus Labrador Hernandez, Ana Patricia Legido Morán, Ricardo León Fernández, Alejandro, Leon-Andrino, Almudena, Llorente, Maria Lopez Pais, Mauricio Loucel Bellino, Sergio, Martin, Miguel Angel Martin-Ferrero, Gonzalo Martinez Municio, Elvira Mateos Alvarez, José María Matilla, Luis María Merino Peñacoba, Mario, Montes-Manrique, Fernando Natal Álvarez, David, C Noriega, Henar Nuñez Del Barrio, Francisco Javier Ortiz de Solórzano Aurusa, Lucía, Pañeda, Victoria Pascual Escudero, Miguel Pascual Samaniego, Adela, Pereda, Marta Pérez Febles, Laura, Pesquera, Maria, Plata, Emma Puertas Ruiz, Mario, Rodriguez-Lopez, Alejandro Romero de Diego, Ana, Ruano, María Ruiz Soriano, Cristina Sánchez Torralvo, Silvia Santiago Maniega, Bárbara, Segura-Méndez, Clarisa Simon Perez, José, Soro-García, Paula Suárez Mansilla Suárez-Mansilla, Jeancarlos Trujillo Díaz, Mª Esther Valsero Herguedas, Andrea Vazquez Fernandez, Eduardo Velasco García, Ana, Zabalza, Álvaro Zamora Horcajada, Fernando Acebes García, Enrique Asensio Díaz, Martín, Bailón, Alejandro David Bueno Cañones, Alvaro Centeno Velasco, Ekta Choolani Bhojwani, Fernando Labarga Rodríguez, Sara, Mambrilla, Pablo, Marcos-Santos, Toledano, Miguel, David Pacheco Sánchez, Baltasar, Pérez-Saborido, Pilar, Pinto, Katherine, Plua, Javier Sanchez Gonzalez, Vicente, Simo, Rosalía Velasco López, Sandra Veleda Belanche, Fabiola Eugenia Michel Campos, Iciar Muñoz Lindez, Maria, Balluerca, Maite, Camuera, Pablo, Helguera, Imanol, Herrero, Patricia Paunero Vazquez, María, Sánchez-Rubio, Isami, Soeda, María Dolores Arribas Del Amo, Vicente Manuel Borrego Estella, Estefania, Casas, Néstor Castán Villanueva, Daniel Delfau Lafuente, Elena Delgado Blanco, María, Domingo, Ismael Gil Romea, Azucena, Gonzalo, Emilio Lagunas Lostao, Noelia Lete Aguirre, Yaiza Martinez Lahoz, Jesús Víctor Pérez-Tierra Ruiz, Maria Del Mar Soriano, Isabel Valero Lázaro, Esmeralda Carnicer Escusol, Guillermo Pola Bandres, Juan Luis Blas Laina, Beatriz, Cros, Patricio Andrés Freile Pazmiño, David, Garcia-Aguilera, Ana Mejía Casado, Ana, Nogués, Issa Talal El-Abur, Carlos, Yánez, Isabel Gascon Ferrer, Carlos, Gracia-Roche, Ursula María Jariod Ferrer, María Victoria Simón Sanz, Tomas, Uson, Waseem, Ahamed, Sumudu, Jayasinghe, Eranda, Karunadasa, Prathibha, Nelihela, Sujeewa Priyantha Bandara Thalgaspitiya, Dickson, Wickramarathna, Nagenthiram, Harivallavan, Pirashanthan, Nagarasa, Eranga, Perera, Kanapathipillai, Piratheep, Vithranage Srimantha Dewsiri Rodrigo, Meera, Thayalan, Rajendram, Thayaparren, Arulprashanth, Arulanantham, Kanishka De Silva, Gagana, Ganga, Anuja Nadeeshan Kumarasinghe, Mahanada, Udukala, Koculen, Vimaleswaran, Prashanthan, Yogarajah, Ravindri, Jayasinghe, Probhodana, Ranaweera, Mohamed, Rishard, Dharmabandhu, N Samarasekera, Sanjeewa, Seneviratne, Sivasuriya, Sivaganesh, Chamal, Fernando, Nishantha, Mendis, Balasingam, Balagobi, Balasupramaniam, Sathesan, Lanka, Dasanayake, Malitha, Patabendige, Wickramasooriya, Dj, Kugarajh, Ganeshapillai, Sathyamoorthy, Prasanna, Dileepa, Ediriweera, Sumudu, Kumarage, Asanka, Jayawardane, Athula, Kaluarachchi, Pituwala Liyanage Adithya Sirisena, Abubakr, Abubakr, Hozifa, Mohamed, Ismail, Mustafa, Mogahid, Sharfeldein, Samir, Ismail, Reem, Musa, Monira, Sarih, Mussab, Abaker, Eman Adam Abdalla, Mohamad, Abdelbagi, Alsafe, Adlan, Ahmed, Alnaeem, Safa, Atyah, Yasir, Awad, Rsheeda, Dawelbait, Mohamed El Hag, Ibrahim, Elbashir, Sami, Eldirdiri, Samwal, Eldirdiri, Mohammed Eltahier Abdalla Omer, Lana Abdalgadir Ahmed Mohamed, Hiba Hassan Rehmtallah Ahmed, Mohammed Balla Yousif Balla, Abdulmoiz, Aljafari, Abdulmalek, Aljafari, Elsagad, Mohamed, Abdulkader, Mohammad, Abdulkader, Shaar, Mohamed Nafea Shaar, Maab Hussein Yousif Elhafiz, Mohammed Husien Yosif Elhafiz, Abubakr Ali Mohammed Alhassan Humidan, Almigdad, Ali, Hisham, Eljack, Mohamedyasin, Elrashid, Manhal, Ahmed, Albrra, Alhag, Douaa, Ali, Mohammed Ahmed Babikir, Imad, Bakheit, Hytham K, S Hamid, Sami, Mohamed, Walid, Shaban, Nidhal, Siddig, Mohamed, Suliman, Rihab Yasir Abdelmagid, Abubaker, Yassin, Omer, Abbas, Eltahir, Abdelrahim, Muhammed, Ahmed, Mohamed, Elobaid, Adil, Hashim, Ali, Mohamed, Omer, Mohamed, Khalid, Osman, Abdallah, Yousif, Safaa, Abdulaal, Marwa, Elsadig, Mohammed, Hussain, Malaz Amir Ataalmanan, Doha Amir AtaAlmanan, Tebyan Amir AtaAlmanan, Alaa, Elgaili, Maria, Mohamed, Usra, Omara, Alkhansa, Osman, Amna, Osman, Safaa, Saad, Abdelkareem, Ahmed, Ali Adil Ali Karar, Aya, Alhassan, Mohanad, Alsidig, Mohamed, Eljack, Mohammed Ibrahim Mohammed Ali, Awad Kamal Awad Osman, Liena, Mohamed, Samah Ibrahim Omer Mohamed Osman Mohamed Osman, Altayeb, Mohammed, Ayman, Mohammed, Khadija, Muhmmed, Eltahir, Musa, Egbal Sahal Abdelmajed, Khalid Z M, Y Hamid, Omnia M, M Yousif, Mahmoud, Saleh, Osman, Bashir, Yassein, Elhussein, Omar, Madani, Eva, Angenete, Jennifer, Park, Lars, Gillberg, Monir, Jawad, Linas, Pieteris, Yousif, Saeed, Sergej, Safonov, Martin, Spångfors, Henrik, Andersson, Hans, Bahlmann, Karin Björnström Karlsson, Michelle, Chew, Lara, Daham, Lina De Geer, Helen, Didriksson, Gunilla, Gagnö, Carina, Jonsson, Anna, Oscarsson, Pamela, Buchwald, Mathilde, Delorme, Marie-Louise, Lydrup, Shahin, Mohseni, Arvid, Pourlotfi, Souheil, Reda, Kajsa, Anderin, Nina, Blomme, Josefin, Segelman, Malin, Simlund, Anders, Thorell, Fredrik, Wogensen, Daniel, Danielsson, Peter, Elbe, Rebecka, Hultgren, Boel, Hynning, Fredrik, Klevebro, Karin, Lind, Ebba, K Lindqvist, Carl, Montan, Ernesto, Sparrelid, Clara Svenberg Lind, Jeremy, Wales, Andreas, Älgå, Martin, Nordberg, Emil, Pieniowski, Ioannis, Gkekas, Niklas, Löfgren, Karolina, Niska, Robert, Zürner, Silvio, Däster, Otto, Kollmar, Savas, Soysal, Stephanie, Taha-Mehlitz, Athanasios, Tampakis, Dominik Valentin Flury, Marie, Heyne-Pietschmann, Gregor Jan Kocher, Ana, M Calinescu, Barbara, Wildhaber, Markus, Gass, Jürg, Metzger, Andreas, Scheiwiller, Lukas, Briner, Oliver, Dwidar, Steven, Grandjean, Pietro, Ricciardi, Alexandra, Calmels, Christodoulou, Michel, Morena, Antonilli, Dimitrios, Christoforidis, Alessandra, Cristaudi, Jacopo, Galafassi, Maria Luisa Gasparri, Francesco, Mongelli, Martino, Munini, Andrea, Papadia, Sotirios Georgios Popeskou, Alex, Alfieri, Thomas, Bächler, Matteo, Giardini, Christian, Gingert, Laura, Guglielmetti, Inna, Meyer, Tobias, Müller, Daniel, Schöni, Folco, Solimene, Frank, Weidner, Marco, Bueter, Daniel, Gero, Amelie, Müller, Carmen, Portenkirchner, Andreas, Thalheimer, Jeannette, Widmer, Ahmad, Hmaideh, Alaa, Kour, Saleem, Swaes, Omar, Alannaz, Ramez, Aljasem, Ezeddin, Dabbagh, M Wafa Hamoud Alhussein, Amr, Hamza, Mazen, Mohammad, Abd El-Fattah Mouhandes, Mohamad, Shareeda, Mohammad Nour Shashaa, Mousa, Sifat, Sheraa, Tahhan, Rahaf, Toutounji, Mohamed, Abdulrahman, Albaraa, Abdulsalam, Ziad Al Jarad, Hala, Alayyoubi, Dina, Alfarra, Ahmad, Alhamid, Mahmoud, Alnasser, Mulham, Gharib, Ruwaid, Gharib, Obaida, Kabel, Osama, Kherallah, Abdulqader, Klaho, Mohamed Shukri Najjar, Ahmad, Razouk, Joud, Shiekhoni, Wael, Alkhaleel, Amira, Almosa, Mohammad, Aloulou, Maen, Alzaeem, Khaled, Arnaout, Muhamad Zakaria Brimo Alsaman, Mohamad, Estanbouli, Hanadi, Hawa, Ahmad Nbrass Kabawi, Ghina, Maarawi, Yehia, Mashhadany, Sana Shaikh Torab, Waleed, Abd, Tayma Abd Alghafour, Hani, Abdalnour, Nazeeh Al Aktaa, Marwan Al Aliwy, M Maher Al Arje, Tasneem Al Najjar, Ahmad, Al-Mouakeh, Ahmad, Aldakhil, Aos, Alhamid, Hasan, Alhasan, Mohamad Shadi Alkarrash, Jumaa, Alkhamis, Sara, Alshekh, Ahmad Yamen Arnaout, Oula, Azizeh, Mohammed, Dalaleh, Aghyad, Danial, Zain, Douba, Raghad, Ghadri, Mohammad Maen Ghannam, Anas, Ghawi, Luma Haj Kassem, M Yasser Halwani, Shahed, Hamadieh, Mohammad, Hanino, Ala Aldeen Hasan, Haya, Jawish, Nasr Ullah Kabbani, Saadullah, Kabbany, Mohamad, Kadi, Lama, Kadoura, Ahmad Amir Kayali, Ammar, Kayyali, Aya, Kelzia, Amana, Kezze, Abdullah, Khalaf, Maymona, Khayata, Mohammad Nour Kitaz, Mohammad, Kour, Raghad, Makki, Ruqaya, Masri, Joseph, Masri, Noor, Masri, Muhammad, Mazketly, Shiar, Mustafa, Ezzeldin, Nashed, Yaman Qoudra Danial, Hasan, Raslan, Rima, Salem, Mohammed Asaad Salem, Thabet Sayed Bakir, Manaf, Sharabaji, Majd Sheikh Alganameh, Duaa Sheikh Kadro, Joudy, Shurbatji, Mohammad Noor Sultan, Dana, Sultan, Hala, Swied, Bakri Tarras Jarkas, Tayf, Toutounji, Fadi, Ward, Rama, Zazo, Aya, Zazo, Zainab, Zeino, Mohamed, Alaktaa, Wael, Alsado, Hala, Fares, Nouran, Hawa, Hamza, Ibrahim, Abdulmonem, Kawas, Mourad, Niazi, Bayan, Toutounji, Mohammad, Abdow, Mohamad, Ahmad, Tareq, Ahmad, Rawan Al Azhar, Hiba Al Hage Diab, Rahaf Al Mulke, Mohammad, Al-Jadaan, Ahmad, Alhussein, Zahra, Alnajem, Omar, Alneser, Bayan, Alsaid, Elias, Azar, Lubna, Bakr, Ammar, Hamza, Yara, Houdifa, Sami, Jomaa, Samer, Kefo, Hiba, Mardini, Alaa, Mohammad, Fadi, Rayya, Fatima, Razzouk, Mohannad, Saleh, Anwar, Shamandi, Ammar Youzkatli Alkhatib, Omar Abdul Salam, Dany, Ajami, Mohammad, Alhamid, Ahmad, Alkhaledi, Danny, Hadidi, Massa, Jabra, Mamdouh, Kallas, Antoine, Laktine, Joelle, Naem, Naya, Naoum, Ghena Mohammed Fawaz Ashour, Ahmad, Alhouri, Deema, Fallouh, Ahmad, N Mohammad, Ahmad, Rmman, Ali, Slitin, Christine, Tanos, Ibrahim, Wakim, Hanaa, Zahrawi, Majd, Abouassi, Layth, Abouassi, Ayham, Aboutrab, Basel, Ahmad, Ahmad Mustafa Ahmad, Aya, Ahmad, Afaf, Ahmad, Mohamad, Alabras, Mohammad, Albasheer, Ola, Alkasser, Mhd Khaled Alkasser, Ali, Alkhdor, Marah, Azkoul, Mohammad Karam Chaaban, Haya, Deeb, Basel, Jazieh, Basel, Kouz, Youssef, Kozah, Raied, Mohamad, Ammar, Mohammad, Mhd Zied Nasri, Mohammed Mouaz Shebani, Mohammed, Shwin, Alaa, Sommaq, Amal, Youssef, Tala Al Asadi, Suhad, Alkhateb, Rasha Khalil Alsayyad, Roula, Altom, Mhd Amin Alzabibi, Batoul, Bakkar, Hlma, Ismail, Salma Khadem Alsrouji, Sara, Melhem, Mohamad, Sankari, Mosa, Shibani, Noura, Shujaa, Ibrahim, 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Giuseppe Salamone, Francesca Toia, Massimiliano Tripoli, Nepogodiev, D, Rosati, R, Surgery, Medicine and Pharmacy academic/administration, Basic (bio-) Medical Sciences, Surgical clinical sciences, Faculty of Medicine and Pharmacy, Gastroenterology, Medical Oncology, Clinical sciences, Breda, João, ARD - Amsterdam Reproduction and Development, Paediatric Surgery, AGEM - Amsterdam Gastroenterology Endocrinology Metabolism, AGEM - Endocrinology, metabolism and nutrition, Graduate School, Tytgat Institute for Liver and Intestinal Research, Laura Alberici, Filippo Antonacci, Alessandro Arena, Angela Belvedere, Fabio Bernagozzi, Paolo Bernante, Pietro Bertoglio, Lorenzo Bianchi, Maria Bisulli, Barbara Bonfanti, Safia Boussedra, Jury Brandolini, Crescenzo Cacciapuoti, Stefano Cardelli, Riccardo Casadei, Matteo Cescon, Alessandro Cipolli, Riccardo Cipriani, Alfredo Conti, Luca Contu, Francesco Costa, Niccolo’ Daddi, Eugenia De Crescenzo, Pierandrea De Iaco, Alessandra De Palma, Massimo Del Gaudio, Anna Nunzia Della Gatta, Giampiero Dolci, Giulia Dondi, Matteo Droghetti, Sergio Nicola Forti Parri, Elena Garelli, Chiara Gelati, Giuliana Germinario, Federico A. Giorgini, Carlo Ingaldi, Elio Jovine, Kenji Kawamukai, Antonio Lanci Lanci, Raffaele Lombardi, Maria Elisa Lozano Miralles, Claudio Marchetti, Michele Masetti, Francesco Minni, Daniele Morezzi, Daniele Parlanti, Alice Pellegrini, Anna Myriam Perrone, Anna paola Pezzuto, Marco Pignatti, Gianluigi Pilu, Valentina Pinto, Gilberto Poggioli, Silvana Bernadetta Puglisi, Diego Raimondo, Matteo Ravaioli, Claudio Ricci, Sara Ricciardi, Francesco Ricotta, Roberta Rizzo, Angela Romano, Matteo Rottoli, Riccardo Schiavina, Renato Seracchioli, Matteo Serenari, Margherita Serra, Piergiorgio Solli, Gioia Sorbi, Mario Taffurelli, Marta Tanzanu, Achille Tarsitano, Marco Tesei, Gabriele Vago, Tommaso Violante, Simone Zanotti, Tommaso Frisoni, leonardo solaini, giorgio ercolani, Fruscio, R, Ornaghi, S, Collaborative, Covidsurg, Collaborative, Globalsurg, Palmisano, Silvia, de Manzini, Nicolo', Žundienė, Laura, Dulskas, Audrius, Kuliavas, Justas, Čekauskas, Albertas, Kielaitė-Gulla, Aistė, Poškus, Tomas, Strupas, Kęstutis, Želvys, Arūnas, COVIDSurg Collaborative, GlobalSurg Collaborative, Department of Surgery, HUS Abdominal Center, Simoes, Jff, Li, E, Picciochi, M, Glasbey, Jc, Baiocchi, G, Blanco-Colino, R, Chaudhry, D, Alameer, E, El-Boghdadly, K, Wuraola, F, Ghosh, D, Gujjuri, Rr, Harrison, Em, Lule, H, Kaafarani, H, Khosravi, M, Kronberger, I, Leventoglu, S, Mann, H, Mclean, Ka, Mengistu, Gm, Modolo, Mm, Ntirenganya, F, Norman, L, Outani, O, Pius, R, Pockney, P, Qureshi, Au, Roslani, Ac, Satoi, S, Shaw, C, Bhangu, A, Omar, Om, Ahmed, Wur, Argus, L, Ball, A, Bywater, Ep, Brar, A, Dawson, Be, Duran, I, Elhadi, M, Jones, C, Kamarajah, Sk, Keatley, Jm, Lawday, S, Marson, Ej, Ots, R, Santos, I, Taylor, Eh, Trout, Im, Varghese, C, Venn, Ml, Xu, W, Dajti, I, Gjata, A, Kacimi, Seo, Boccalatte, L, Cox, D, Townend, P, Aigner, F, Kronberger, Ie, Samadov, E, Alderazi, A, Hossain, K, Padmore, G, van Ramshorst, G, Lawani, I, Cerovac, A, Delibegovic, S, Gomes, Gma, Buarque, Il, Gohar, M, Slavchev, M, Nwegbu, C, Agarwal, A, Martin, J, Ng-Kamstra, J, Olivos, M, Lou, Wh, Ren, Dl, Calvache, Ja, Rivera, Cjp, Hadzibegovic, Ad, Kopjar, T, Mihanovic, J, Jimenez, Pma, Gouvas, N, Klat, J, Novysedlak, R, Amisi, N, Christensen, P, El-Hussuna, A, Batista, S, Lincango-Naranjo, E, Emile, S, Sandoval, Daa, Dhufera, H, Hailu, S, Mengesha, Mg, Kauppila, Jh, Arnaud, Ap, Demetrashvili, Z, Albertsmeier, M, Lederhuber, H, Loffler, Mw, Acquah, Dk, Ofori, B, Tabiri, S, Metallidis, S, Tsoulfas, G, Aguilera-Arevalo, Ml, Recinos, G, Mersich, T, Wettstein, D, Kembuan, G, Milan, Pb, Khosravi, Mh, Mozafari, M, Hilmi, A, Mohan, H, Zmora, O, Gallo, G, Pata, F, Pellino, G, Fujimoto, Y, Kuroda, N, Abou Chaar, Mk, Ayasra, F, Fakhradiyev, I, Hamdun, Ih, Jin-Young, J, Jamal, M, Karout, L, Gulla, A, Rasoaherinomenjanahary, F, Samison, Lh, Sanchez, Iid, Gonzalez, D, Martinez, L, Martinez, Mj, Nayen, A, and Ramos-De la Medina, A
Subjects: Male, Vascular damage Radboud Institute for Health Sciences [Radboudumc 16], MULTICENTER, infectious diseases, 0302 clinical medicine, 030202 anesthesiology, Anesthesiology, Cirugía, 80 and over, Prospective Studies, Young adult, Child, Lung, Aged, 80 and over, COVID-19, delay, SARS-CoV-2, surgery, timing, SARS‐CoV‐2 infection, Operative, Child, Preschool, Pneumonia & Influenza, Infection, Cohort study, Human, medicine.medical_specialty, Clinical Sciences, Sars-cov-2, GlobalSurg Collaborative, Settore MED/28 - MALATTIE ODONTOSTOMATOLOGICHE, 03 medical and health sciences, Clinical Research, Biodefense, Correspondence, Humans, PULMONARY COMPLICATIONS, CRIANÇAS EM IDADE PRÉ-ESCOLAR, Aged, Science & Technology, Cirurgia, Prevention, Infant, Odds ratio, Pneumonia, Prospective Studie, Clinical research, Surgery, Human medicine, 610 Medizin und Gesundheit, 1109 Neurosciences, Internationality, Settore MED/29 - CHIRURGIA MAXILLOFACCIALE, Settore MED/18 - CHIRURGIA GENERALE, Adolescent, Adult, Cohort Studies, Female, Middle Aged, Surgical Procedures, Operative, Time, Young Adult, Practice Guidelines as Topic, gastroenterology, Medicine and Health Sciences, 030212 general & internal medicine, Prospective cohort study, Mortality rate, Covid19, Infectious Diseases, Patient Safety, medicine.symptom, 6.4 Surgery, Life Sciences & Biomedicine, 610 Medicine & health, COVID-19, SARS-CoV-2, delay, surgery, timing, Asymptomatic, NO, COVIDSurg Collaborative, Vaccine Related, All institutes and research themes of the Radboud University Medical Center, medicine, Mortalitat, ddc:610, Mortality, Preschool, LS7_4, Surgical Procedures, business.industry, Neurosciences, Evaluation of treatments and therapeutic interventions, 1103 Clinical Sciences, 3126 Surgery, anesthesiology, intensive care, radiology, Reconstructive and regenerative medicine Radboud Institute for Health Sciences [Radboudumc 10], Anesthesiology and Pain Medicine, Emerging Infectious Diseases, Good Health and Well Being, Mortalidad, Cohort Studie, business
Abstract: Peri-operative SARS-CoV-2 infection increases postoperative mortality. The aim of this study was to determine the optimal duration of planned delay before surgery in patients who have had SARS-CoV-2 infection. This international, multicentre, prospective cohort study included patients undergoing elective or emergency surgery during October 2020. Surgical patients with pre-operative SARS-CoV-2 infection were compared with those without previous SARS-CoV-2 infection. The primary outcome measure was 30-day postoperative mortality. Logistic regression models were used to calculate adjusted 30-day mortality rates stratified by time from diagnosis of SARS-CoV-2 infection to surgery. Among 140,231 patients (116 countries), 3127 patients (2.2%) had a pre-operative SARS-CoV-2 diagnosis. Adjusted 30-day mortality in patients without SARS-CoV-2 infection was 1.5% (95%CI 1.4-1.5). In patients with a pre-operative SARS-CoV-2 diagnosis, mortality was increased in patients having surgery within 0-2 weeks, 3-4 weeks and 5-6 weeks of the diagnosis (odds ratio (95%CI) 4.1 (3.3-4.8), 3.9 (2.6-5.1) and 3.6 (2.0-5.2), respectively). Surgery performed ≥ 7 weeks after SARS-CoV-2 diagnosis was associated with a similar mortality risk to baseline (odds ratio (95%CI) 1.5 (0.9-2.1)). After a ≥ 7 week delay in undertaking surgery following SARS-CoV-2 infection, patients with ongoing symptoms had a higher mortality than patients whose symptoms had resolved or who had been asymptomatic (6.0% (95%CI 3.2-8.7) vs. 2.4% (95%CI 1.4-3.4) vs. 1.3% (95%CI 0.6-2.0), respectively). Where possible, surgery should be delayed for at least 7 weeks following SARS-CoV-2 infection. Patients with ongoing symptoms ≥ 7 weeks from diagnosis may benefit from further delay. ispartof: ANAESTHESIA vol:76 issue:6 pages:748-758 ispartof: location:England status: published
Published: 2021

39. Genomic epidemiology reveals transmission patterns and dynamics of SARS-CoV-2 in Aotearoa New Zealand

Author: James Hadfield, Matthew Storey, Nigel P. French, Fábio K. Mendes, Andrew Sporle, Sue Huang, David Welch, Shevaun Paine, Xiaoyun Ren, Jemma L. Geoghegan, Colin R Simpson, Michael G Baker, Sebastián Duchêne, Sarah Jefferies, David R. Murdoch, Joep de Ligt, Edward C. Holmes, Lauren Jelley, Jordan Douglas, Alexei J. Drummond, and Jill Sherwood
Subjects: SARS-CoV-2/classification, 0106 biological sciences, 0301 basic medicine, Male, Epidemiology, General Physics and Astronomy, 01 natural sciences, law.invention, law, Pandemic, New Zealand/epidemiology, Child, Phylogeny, COVID-19/epidemiology, Aged, 80 and over, education.field_of_study, Multidisciplinary, Geography, Genomics, Middle Aged, Aotearoa, Phylogenetics, Transmission (mechanics), Child, Preschool, Female, Adult, medicine.medical_specialty, Lineage (genetic), Adolescent, Science, Population, Genome, Viral, Disease cluster, 010603 evolutionary biology, Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Young Adult, Whole Genome Sequencing/methods, Quarantine, medicine, Humans, Genome, Viral/genetics, education, Pandemics, Aged, Whole Genome Sequencing, SARS-CoV-2, Public health, Infant, Newborn, COVID-19, Infant, General Chemistry, Genomics/methods, Genome evolution, 030104 developmental biology, Demography, New Zealand
Abstract: New Zealand, a geographically remote Pacific island with easily sealable borders, implemented a nationwide ‘lockdown’ of all non-essential services to curb the spread of COVID-19. Here, we generate 649 SARS-CoV-2 genome sequences from infected patients in New Zealand with samples collected during the ‘first wave’, representing 56% of all confirmed cases in this time period. Despite its remoteness, the viruses imported into New Zealand represented nearly all of the genomic diversity sequenced from the global virus population. These data helped to quantify the effectiveness of public health interventions. For example, the effective reproductive number, Re of New Zealand’s largest cluster decreased from 7 to 0.2 within the first week of lockdown. Similarly, only 19% of virus introductions into New Zealand resulted in ongoing transmission of more than one additional case. Overall, these results demonstrate the utility of genomic pathogen surveillance to inform public health and disease mitigation., New Zealand implemented stringent COVID-19 control measures early after identification of its first case. Here, the authors perform whole genome sequencing of samples taken until 22 May 2020 and find high viral diversity indicative of multiple separate introductions and limited community transmission.
Published: 2020
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40. The power and limitations of genomics to track COVID-19 outbreaks: a case study from New Zealand

Author: Xiaoyun Ren, Olin K. Silander, Jordan Douglas, Andrew Sporle, Shevaun Paine, Lauren Jelley, Joep de Ligt, Sue Huang, David R. Murdoch, Alexei J. Drummond, David Welch, Jemma L. Geoghegan, Matt Storey, Sarah Jefferies, Nikki E. Freed, Jillian Sherwood, Edward C. Holmes, Nigel P. French, James Hadfield, Michael G Baker, and Colin R Simpson
Subjects: Economic growth, Geography, Single cluster, Coronavirus disease 2019 (COVID-19), Research council, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Outbreak, Sample (statistics), Genomics, Track and trace
Abstract: SummaryBackgroundReal-time genomic sequencing has played a major role in tracking the global spread and local transmission of SARS-CoV-2, contributing greatly to disease mitigation strategies. After effectively eliminating the virus, New Zealand experienced a second outbreak of SARS-CoV-2 in August 2020. During this August outbreak, New Zealand utilised genomic sequencing in a primary role to support its track and trace efforts for the first time, leading to a second successful elimination of the virus.MethodsWe generated the genomes of 80% of the laboratory-confirmed samples of SARS-CoV-2 from New Zealand’s August 2020 outbreak and compared these genomes to the available global genomic data.FindingsGenomic sequencing was able to rapidly identify that the new COVID-19 cases in New Zealand belonged to a single cluster and hence resulted from a single introduction. However, successful identification of the origin of this outbreak was impeded by substantial biases and gaps in global sequencing data.InterpretationAccess to a broader and more heterogenous sample of global genomic data would strengthen efforts to locate the source of any new outbreaks.FundingThis work was funded by the Ministry of Health of New Zealand, New Zealand Ministry of Business, Innovation and Employment COVID-19 Innovation Acceleration Fund (CIAF-0470), ESR Strategic Innovation Fund and the New Zealand Health Research Council (20/1018 and 20/1041).
Published: 2020

41. Characterising the epidemic spread of influenza A/H3N2 within a city through phylogenetics

Author: Christian H. Nickel, Andreas Zeller, Myrta Brunner, Daniela Lang, Michael Naegele, Tanja Stadler, Olivier Dubuis, Helena M. B. Seth-Smith, James Hadfield, Trevor Bedford, Andreas Buser, Daniel Wüthrich, Mohammedyaseen Syedbasha, Richard A. Neher, Rita Schneider-Sliwa, Nicole Ritz, Nicola F. Müller, Nadine Sailer, Yvonne Hollenstein, Adrian Egli, Nina Goldman, Brian M. Lang, Noémi Augustin, Claudia Saalfrank, and Manuel Battegay
Subjects: Viral Diseases, Atmospheric Science, Psychological intervention, Social Sciences, law.invention, Disease Outbreaks, 0302 clinical medicine, Medical Conditions, Sociology, law, Pandemic, Medicine and Health Sciences, 030212 general & internal medicine, Biology (General), Child, Phylogeny, Data Management, 0303 health sciences, Schools, Phylogenetic Analysis, 3. Good health, Phylogenetics, Geography, Transmission (mechanics), Infectious Diseases, Child, Preschool, Evolutionary Rate, Seasons, Switzerland, Research Article, Computer and Information Sciences, Evolutionary Processes, Adolescent, QH301-705.5, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Education, 03 medical and health sciences, Meteorology, Epidemic spread, Influenza, Human, Humans, Evolutionary Systematics, Cities, Epidemics, 030304 developmental biology, Taxonomy, Evolutionary Biology, Influenza A Virus, H3N2 Subtype, Outbreak, Biology and Life Sciences, Influenza a, Humidity, RC581-607, Influenza, Health Care, Transmission network, Age Groups, People and Places, Earth Sciences, Population Groupings, Geriatric Care, Immunologic diseases. Allergy, Demography
Abstract: Infecting large portions of the global population, seasonal influenza is a major burden on societies around the globe. While the global source sink dynamics of the different seasonal influenza viruses have been studied intensively, its local spread remains less clear. In order to improve our understanding of how influenza is transmitted on a city scale, we collected an extremely densely sampled set of influenza sequences alongside patient metadata. To do so, we sequenced influenza viruses isolated from patients of two different hospitals, as well as private practitioners in Basel, Switzerland during the 2016/2017 influenza season. The genetic sequences reveal that repeated introductions into the city drove the influenza season. We then reconstruct how the effective reproduction number changed over the course of the season. While we did not find that transmission dynamics in Basel correlate with humidity or school closures, we did find some evidence that it may positively correlated with temperature. Alongside the genetic sequence data that allows us to see how individual cases are connected, we gathered patient information, such as the age or household status. Zooming into the local transmission outbreaks suggests that the elderly were to a large extent infected within their own transmission network. In the remaining transmission network, our analyses suggest that school-aged children likely play a more central role than pre-school aged children. These patterns will be valuable to plan interventions combating the spread of respiratory diseases within cities given that similar patterns are observed for other influenza seasons and cities., Author Summary As shown with the current SARS-CoV-2 pandemic, respiratory diseases can quickly spread around the globe. While it can be important to understand how diseases spread around the globe, local spread is most often the main driver of novel infections of respiratory diseases such as SARS-CoV-2 or influenza. We here use genetic sequence data alongside patient information to better understand what drives the local spread of influenza by looking at the 2016/2017 influenza season in Basel, Switzerland as an example. The genetic sequence data allows us to reconstruct how the transmission dynamics changed over the course of the season, which we then compare to trends in humidity and temperature and times when schools were open or closed. Additionally, the genetic sequence data allows us to see how individual cases are connected. Using patient information, such as age and household status our analyses suggest that the elderly mainly transmit within their own transmission network. Additionally, they suggest that school aged children, but not necessarily pre-school aged children are important drivers of the local spread of influenza.
Published: 2020

42. 60 Development and validation of blood tumor mutational burden reference standards

Author: Eun-Ang Rabier-Moreau, James Hadfield, Guillem Portella, Matthew James Butler, and Yves Konigshofer
Subjects: Oncology, medicine.medical_specialty, Calibration and validation, business.industry, Mutant allele, Fragment size, Lymphoblastoid cell, Internal medicine, Blood tumor, medicine, In patient, business, Reference standards, Exome
Abstract: Background Tumor mutational burden (TMB), as measured by exome or panel sequencing of tumor tissue (tTMB) or blood (bTMB), has been identified as a potential predictive biomarker for treatment benefit in patients with various cancer types receiving immunotherapy targeting checkpoint inhibitors (e.g. PD-1, PD-L1, CTLA-4). However, significant variability in TMB measurement has been reported due to differences in pre-analytical and laboratory methods, panel size, number of genes covered and bioinformatics pipelines. Reference standards have been proposed and evaluated for tTMB analysis by the Friends of Cancer Research (FoCR) to enable harmonization and standardization across different tTMB panel providers. Reference standards for bTMB are likely to be even more important given the unique challenges and higher sensitivity required for bTMB assays. Methods Contrived bTMB reference materials with 0.5% and 2% tumor content were developed using DNA from tumor cell lines and donor-matched lymphoblastoid cell lines fragmented and size-selected to mimic cell-free DNA with TMB scores of 7, 9, 20 and 26 mut/Mb. Mutation coverage, mutant allele frequency (MAF) and bTMB scores were assessed using the PredicineATLAS and GuardantOMNI next-generation sequencing (NGS) platforms. Results The DNA fragment size for the contrived samples was similar to naturally occurring circulating cell-free tumor DNA and mutation patterns were aligned with those from parental tumor lines. As anticipated, low frequency artefactual MAF variants were observed, requiring removal by bioinformatic filtration. For samples with 2% tumor content, standards for 7, 20 and 26 mut/Mb were found to have as-expected bTMB scores across both evaluation platforms, with good reproducibility, following removal of low frequency MAFs. Results for 0.5% tumour content were also promising, although with greater variability in post-filtration bTMB scores observed. Conclusions The findings demonstrate it is feasible to produce bTMB reference standards across a range of bTMB levels. The data highlight the importance of data filtration to account for underlying low MAFs in such cell-line derived samples and that this reference material can control for variant sensitivity though not variant specificity. bTMB reference standards reported here could support the calibration and validation of bTMB platforms and help harmonization between panels and laboratories, thus improving the accuracy of testing to aid treatment decisions in oncology. Acknowledgements The study was funded by AstraZeneca. Medical writing support, which was in accordance with Good Publication Practice (GPP3) guidelines, was provided by Rachel Cicchelli, PhD, of Cirrus Communications (Macclesfield, UK), an Ashfield company, and was funded by AstraZeneca.
Published: 2020

43. Viral genome sequencing places White House COVID-19 outbreak into phylogenetic context

Author: Jennifer K. Logue, Trevor Bedford, Jay Shendure, James Hadfield, Helen Y. Chu, Erica Ryke, Peter D Han, Mark J. Rieder, Chris D. Frazar, Caitlin R Wolf, Jover Lee, Benjamin Pelle, Lea M. Starita, Deborah A. Nickerson, and Christina M. Lockwood
Subjects: White (mutation), medicine.medical_specialty, Geography, Phylogenetic tree, Lineage (evolution), Epidemiology, medicine, Zoology, Outbreak, Context (language use), Genome, DNA sequencing
Abstract: In October 2020, an outbreak of at least 50 COVID-19 cases was reported surrounding individuals employed at or visiting the White House. Here, we applied genomic epidemiology to investigate the origins of this outbreak. We enrolled two individuals with exposures linked to the White House COVID-19 outbreak into an IRB-approved research study and sequenced their SARS-CoV-2 infections. We find these viral sequences are identical to each other, but are distinct from over 190,000 publicly available SARS-CoV-2 genomes. These genomes fall as part of a lineage circulating in the USA since April or May 2020 and detected in Virginia and Michigan. Looking forwards, sequencing of additional community SARS-CoV-2 infections collected in the USA prior to October 2020 may shed further light on its geographic ancestry. In sequencing of SARS-CoV-2 infections collected after October 2020, it may be possible to identify infections that likely descend from the White House COVID-19 outbreak.
Published: 2020

44. A phylodynamic workflow to rapidly gain insights into the dispersal history and dynamics of SARS-CoV-2 lineages

Author: Tony Wawina-Bokalanga, Maria Artesi, Mandev S. Gill, Simon Dellicour, Samuel L. Hong, Marius Gilbert, Joan Martí-Carreras, Vincent Bours, Piet Maes, Nuno R. Faria, Sébastien Bontems, Conor R Walker, James Hadfield, Guy Baele, Emmanuel André, Cécile Meex, Marie-Pierre Hayette, Keith Durkin, Bert Vanmechelen, Nicola De Maio, Medical Research Council-São Paulo Research Foundation (FAPESP), and Wellcome Trust
Subjects: Letter, Coronavirus disease 2019 (COVID-19), Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Physical Distancing, phylogenetic clusters, Genome, Viral, Biology, phylogeography, 0601 Biochemistry and Cell Biology, AcademicSubjects/SCI01180, Workflow, Evolution, Molecular, Patient Isolation, 03 medical and health sciences, Spatio-Temporal Analysis, 0302 clinical medicine, lockdown measures, 0603 Evolutionary Biology, Belgium, Intervention measures, phylodynamic, Pandemic, Genetics, Humans, 030212 general & internal medicine, Molecular Biology, Ecology, Evolution, Behavior and Systematics, Phylogeny, 030304 developmental biology, Spatial density, Evolutionary Biology, 0604 Genetics, Likelihood Functions, 0303 health sciences, SARS-CoV-2, AcademicSubjects/SCI01130, Genetic data, COVID-19, Genomics, Sciences bio-médicales et agricoles, Pipeline (software), Geography, Proof of concept, Evolutionary biology, Mutation, Biological dispersal, 030217 neurology & neurosurgery
Abstract: Since the start of the COVID-19 pandemic, an unprecedented number of genomic sequences of SARS-CoV-2 have been generated and shared with the scientific community. The unparalleled volume of available genetic data presents a unique opportunity to gain real-time insights into the virus transmission during the pandemic, but also a daunting computational hurdle if analyzed with gold-standard phylogeographic approaches. To tackle this practical limitation, we here describe and apply a rapid analytical pipeline to analyze the spatiotemporal dispersal history and dynamics of SARS-CoV-2 lineages. As a proof of concept, we focus on the Belgian epidemic, which has had one of the highest spatial densities of available SARS-CoV-2 genomes. Our pipeline has the potential to be quickly applied to other countries or regions, with key benefits in complementing epidemiological analyses in assessing the impact of intervention measures or their progressive easement., SCOPUS: ar.j, info:eu-repo/semantics/published
Published: 2020

45. The Seattle Flu Study: a multiarm community-based prospective study protocol for assessing influenza prevalence, transmission and genomic epidemiology

Author: Seattle Flu Study Investigators, Victoria Lyon, Jay Shendure, Reena Gulati, Janet A. Englund, Trevor Bedford, James Hadfield, Kirsten Lacombe, Thomas R. Sibley, Michael L. Jackson, Michael Famulare, Amanda L. Adler, Barry R. Lutz, Helen Y. Chu, Co-Investigators, Caitlin R Wolf, Kira L. Newman, Peter D Han, Michael Boeckh, Jennifer Logue, Lea M. Starita, Louise E. Kimball, Monica Zigman Suchsland, Anahita Kiavand, Elisabeth Brandstetter, Deborah A. Nickerson, Mark J. Rieder, and Chris D. Frazar
Subjects: medicine.medical_specialty, Epidemiology, Population, International airport, Ambulatory care, respiratory infection, Environmental health, Influenza, Human, Pandemic, Prevalence, medicine, Humans, protocol, Prospective Studies, Prospective cohort study, education, Protocol (science), education.field_of_study, Transmission (medicine), business.industry, Respiratory infection, General Medicine, Genomics, Institutional review board, Metropolitan area, virology, surveillance, Medicine, Seasons, influenza, business
Abstract: IntroductionInfluenza epidemics and pandemics cause significant morbidity and mortality. An effective response to a potential pandemic requires the infrastructure to rapidly detect, characterize, and potentially contain new and emerging influenza strains at a population level. The objective of this study is to use data gathered simultaneously from community and hospital sites to develop a model of how influenza enters and spreads in a population.Methods and AnalysisStarting in the 2018-19 season, we have been enrolling individuals with acute respiratory illness from community sites throughout the Seattle metropolitan area, including clinics, childcare facilities, Seattle-Tacoma International Airport, workplaces, college campuses, and homeless shelters. At these sites, we collect clinical data and mid-nasal swabs from individuals with at least two acute respiratory symptoms. Additionally, we collect residual nasal swabs and data from individuals who seek care for respiratory symptoms at four regional hospitals. Samples are tested using a multiplex molecular assay, and influenza whole genome sequencing is performed for samples with influenza detected. Geospatial mapping and computational modeling platforms are in development to characterize the regional spread of influenza and other respiratory pathogens.Ethics and DisseminationThe study was approved by the University of Washington’s Institutional Review Board. Results will be disseminated through talks at conferences, peer-reviewed publications, and on the study website (www.seattleflu.org).Article SummaryStrengths and limitations of this study-Large-scale multiple-arm study of respiratory illness characterization with collection of samples from individuals in the community as well as in ambulatory care and hospital settings-Integration of sociodemographic, clinical, and geospatial data on a regional level-Multiplex molecular testing for multiple viral and bacterial pathogens and whole genome sequencing of influenza for detailed molecular epidemiologic characterization and transmission mapping-Geographically and socioeconomically diverse sampling of community-based acute respiratory illnesses
Published: 2020

46. Operationalizing genomic epidemiology during the Nord-Kivu Ebola outbreak, Democratic Republic of the Congo

Author: Marceline Akonga, Moussa Moïse Diagne, Steve Ahuka-Mundeke, Amadou A. Sall, Bailey White, Allison Black, Martin Faye, Oumar Faye, Martine Peeters, Bibiche Nsunda, Francois Edidi-Atani, Gustavo Palacios, F. Mambu, Eric Delaporte, Placide Mbala-Kingebeni, Nella Bisento, Catherine B. Pratt, N. Di Paola, Amuri Aziza, James Hadfield, Trevor Bedford, M. Matondo, Daniel Mukadi, Junior Bulabula, Eddy Kinganda-Lusamaki, Jean Jacques Muyembe-Tamfum, and Michael R. Wiley
Subjects: medicine.medical_specialty, Ebola virus, Operationalization, business.industry, media_common.quotation_subject, Environmental resource management, Outbreak, Epidemiologic Surveillance, medicine.disease_cause, Democracy, law.invention, Data sequences, Transmission (mechanics), Geography, law, Epidemiology, medicine, business, media_common
Abstract: The Democratic Republic of the Congo declared its tenth Ebola virus disease outbreak in July 2018, which has circulated primarily in the Nord Kivu province. In addition to standard epidemiologic surveillance and response efforts, the Institut National de Recherche Biomédicale implemented an end-to-end genomic surveillance system, including sequencing, bioinformatic analysis, and dissemination of genomic epidemiologic results to frontline public health workers. Here we report 538 new genomes from this outbreak; together with previously available sequence data (n = 48 genomes), this represents an unprecedented 17% of all laboratory-confirmed infections. To support outbreak response efforts, we reconstructed spatiotemporal transmission dynamics at broad and at fine scales as new data were available and disseminated the results via an interactive narrative-based platform. Our innovative system enables actionable information sharing between scientists and epidemiologists coordinating the day-to-day response on the time scales necessary to guide response efforts. The development of this genomic surveillance pipeline, within a resource-limited setting, represents significant technological and scientific progress in genomic epidemiology. Here we present a phylodynamic analysis of the outbreak as of February 2020, and describe the types of epidemiologic dynamics that we monitor the genomic data for, including resolution of co-circulating transmission chains, detection of superspreading events, inference of regions that act as transmission sources and sinks, and differentiation of closely linked cases versus propagated transmission. These findings have ameliorated the current outbreak response and are directly applicable to future outbreaks.
Published: 2020

47. Visualizing variation within Global Pneumococcal Sequence Clusters (GPSCs) and country population snapshots to contextualize pneumococcal isolates

Author: Shabir A. Madhi, Stephen D. Bentley, Rebecca A. Gladstone, M.A. Du Plessis, Lesley McGee, Pak-Leung Ho, Waleria Hryniewicz, A. von Gottberg, Stephanie W. Lo, Yanping Li, Benjamin Taylor, Benjamin J. Metcalf, Leon J. Bentley, Martin Antonio, Corin A. Yeats, James Hadfield, David M. Aanensen, Keith P. Klugman, Dean Everett, F X Quah, Nicole L Pershing, Carrie L. Byington, Bernard Beall, Paulina A. Hawkins, Robert F. Breiman, Kedibone M. Ndlangisa, Almeida Scg., Jennifer E. Cornick, Anne J. Blaschke, Theresa J. Ochoa, Ron Dagan, Rachel Benisty, John A. Lees, Veeraraghavan Balaji, Andrew J. Pollard, K L Ravikumar, Richard Goater, A J van Tonder, Nicholas J. Croucher, Brenda Kwambana-Adams, Beutel Sigaùque, and Susan A. Nzenze
Subjects: Most recent common ancestor, Serotype, antibiotic resistance, pangenome, Population, Microbial evolution and epidemiology: Population Genomics, Locus (genetics), Biology, purl.org/pe-repo/ocde/ford#3.03.09 [https], Serogroup, Evolution, Molecular, South Africa, BioResource, Phylogenetics, Utah, Databases, Genetic, Drug Resistance, Bacterial, purl.org/pe-repo/ocde/ford#1.06.01 [https], Clade, education, Phylogeny, education.field_of_study, whole genome sequencing, Phylogenetic tree, phylogenetic dating, Polysaccharides, Bacterial, High-Throughput Nucleotide Sequencing, population structure, General Medicine, Sequence Analysis, DNA, pneumococcal, recombination, Phylogeography, Streptococcus pneumoniae, Composite transposon, Evolutionary biology, DNA Transposable Elements, Poland, purl.org/pe-repo/ocde/ford#1.06.07 [https]
Abstract: Knowledge of pneumococcal lineages, their geographic distribution and antibiotic resistance patterns, can give insights into global pneumococcal disease. We provide interactive bioinformatic outputs to explore such topics, aiming to increase dissemi-nation of genomic insights to the wider community, without the need for specialist training. We prepared 12 country-specific phylogenetic snapshots, and international phylogenetic snapshots of 73 common Global Pneumococcal Sequence Clusters (GPSCs) previously defined using PopPUNK, and present them in Microreact. Gene presence and absence defined using Roary, and recombination profiles derived from Gubbins are presented in Phandango for each GPSC. Temporal phylogenetic signal was assessed for each GPSC using BactDating. We provide examples of how such resources can be used. In our example use of a country-specific phylogenetic snapshot we determined that serotype 14 was observed in nine unrelated genetic backgrounds in South Africa. The international phylogenetic snapshot of GPSC9, in which most serotype 14 isolates from South Africa were observed, highlights that there were three independent sub-clusters represented by South African serotype 14 isolates. We estimated from the GPSC9-dated tree that the sub-clusters were each established in South Africa during the 1980s. We show how recombination plots allowed the identification of a 20 kb recombination spanning the capsular polysaccharide locus within GPSC97. This was consistent with a switch from serotype 6A to 19A estimated to have occured in the 1990s from the GPSC97-dated tree. Plots of gene presence/absence of resistance genes (tet, erm, cat) across the GPSC23 phylogeny were consistent with acquisition of a composite transposon. We estimated from the GPSC23-dated tree that the acquisition occurred between 1953 and 1975. Finally, we demonstrate the assignment of GPSC31 to 17 externally generated pneumococcal serotype 1 assemblies from Utah via Pathogenwatch. Most of the Utah isolates clustered within GPSC31 in a USA-specific clade with the most recent common ancestor estimated between 1958 and 1981. The resources we have provided can be used to explore to data, test hypothesis and generate new hypotheses. The accessible assignment of GPSCs allows others to contextualize their own collections beyond the data presented here.
Published: 2020

48. Cryptic transmission of SARS-CoV-2 in Washington State

Author: Jay Shendure, Kirsten Lacombe, Xianding Deng, Suxiang Tong, Richard A. Neher, Scot Federman, Michael Famulare, Jover Lee, Melissa Truong, Caitlin R Wolf, Geoffrey S. Baird, Matthew Richardson, Amanda Adler, Charles Y. Chiu, Scott Lindquist, Thomas R. Sibley, Geoff Melly, Elisabeth Brandstetter, Deborah A. Nickerson, Duncan MacCannell, Anahita Kiavand, Krista Queen, Pavitra Roychoudhury, Janet A. Englund, Gregory Pepper, Wei Gu, Lasata Shrestha, Ying Tao, Emma B. Hodcroft, Trevor Bedford, Danielle Giroux, Nicola F. Müller, Kairsten Fay, Lea M. Starita, Misja Ilcisin, Philip Dykema, Jeffrey S. Duchin, Louise H. Moncla, Alexander L. Greninger, Shari Cho, Hong Xie, James Hadfield, Brian Hiatt, Keith R. Jerome, Peter D Han, Arun K. Nalla, Gregory L. Armstrong, Helen Y. Chu, Anna Uehara, Truong N. Nguyen, Romesh Gautom, John Huddleston, Adam Reinhardt, Meei-Li Huang, Mark J. Rieder, and Chris D. Frazar
Subjects: Washington, medicine.medical_specialty, 2019-20 coronavirus outbreak, Coronavirus disease 2019 (COVID-19), Epidemiology, Evolution, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Pneumonia, Viral, Genomics, Genome, Viral, World health, Article, Betacoronavirus, 03 medical and health sciences, 0302 clinical medicine, Report, Pandemic, medicine, Humans, 030212 general & internal medicine, Clade, Pandemics, Phylogeny, 030304 developmental biology, Likelihood Functions, 0303 health sciences, Multidisciplinary, biology, SARS-CoV-2, Transmission (medicine), Outbreak, COVID-19, Bayes Theorem, biology.organism_classification, Virology, 3. Good health, Geography, Coronavirus Infections, Reports, Demography
Abstract: A series of unfortunate events The history of how severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spread around the planet has been far from clear. Several narratives have been propagated by social media and, in some cases, national policies were forged in response. Now that many thousands of virus sequences are available, two studies analyzed some of the key early events in the spread of SARS-CoV-2. Bedford et al. found that the virus arrived in Washington state in late January or early February. The viral genome from the first case detected had mutations similar to those found in Chinese samples and rapidly spread and dominated subsequent undetected community transmission. The other viruses detected had origins in Europe. Worobey et al. found that early introductions into Germany and the west coast of the United States were extinguished by vigorous public health efforts, but these successes were largely unrecognized. Unfortunately, several major travel events occurred in February, including repatriations from China, with lax public health follow-up. Serial, independent introductions triggered the major outbreaks in the United States and Europe that still hold us in the grip of control measures. Science, this issue p. 571, p. 564, The first case of SARS-CoV-2 in Washington state occurred at the end of January and, by going undetected, sparked community transmission., After its emergence in Wuhan, China, in late November or early December 2019, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus rapidly spread globally. Genome sequencing of SARS-CoV-2 allows the reconstruction of its transmission history, although this is contingent on sampling. We analyzed 453 SARS-CoV-2 genomes collected between 20 February and 15 March 2020 from infected patients in Washington state in the United States. We find that most SARS-CoV-2 infections sampled during this time derive from a single introduction in late January or early February 2020, which subsequently spread locally before active community surveillance was implemented.
Published: 2020
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49. High-resolution influenza mapping of a city reveals socioeconomic determinants of transmission within and between urban quarters

Author: Yvonne Hollenstein, Nadine Sailer, Annette Blaich, Nina Goldman, Helena M.B. Seth-Smith, Claudia Saalfrank, Myrta Brunner, Mohammedyaseen Syedbasha, Manuel Battegay, Thomas Vogel, Rita Schneider-Sliwa, Olivier Dubuis, Sarah Tschudin-Sutter, Trevor Bedfors, Richard A. Neher, Daniela Lang, Nicola F. Müller, Jan Bauer, Andreas Buser, Nadezhda Amar-Sliwad, Noemie Augustin, Andreas Zeller, Daniel Wuethrich, Christian H. Nickel, Michael Naegele, Tanja Stadler, Nicole Ritz, James Hadfield, Adrian Egli, and Emma B. Hodcroft
Subjects: 0303 health sciences, medicine.medical_specialty, 030306 microbiology, Public health, 1. No poverty, Quarter (United States coin), 3. Good health, law.invention, 03 medical and health sciences, Transmission (mechanics), Geography, law, Net income, Scale (social sciences), 11. Sustainability, Pandemic, medicine, Household income, Socioeconomics, Socioeconomic status, 030304 developmental biology
Abstract: With two-thirds of the global population projected to be living in urban areas by 2050, understanding the transmission patterns of viral pathogens within cities is crucial for effective prevention strategies. Here, in unprecedented spatial resolution, we analysed the socioeconomic determinants of influenza transmission in a European city. We combined geographical and epidemiological data with whole genome sequencing of influenza viruses at the scale of urban quarters and statistical blocks, the smallest geographic subdivisions within a city. We observed annually re-occurring geographic clusters of influenza incidences, mainly associated with net income, and independent of population density and living space. Vaccination against influenza was also mainly associated with household income and was linked to the likelihood of influenza-like illness within an urban quarter. Transmissions patterns within and between quarters were complex. High-resolution city-level epidemiological studies combined with social science surveys such as this will be essential for understanding seasonal and pandemic transmission chains and delivering tailored public health information and vaccination programs at the municipal level.
Published: 2020
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50. The Nature and Extent of Plasmid Variation in Chlamydia trachomatis

Author: Ian N. Clarke, Peter Marsh, Nicholas R. Thomson, David W. Cleary, Charlotte A Jones, James Hadfield, and Colette O'Neill
Subjects: Microbiology (medical), Single-nucleotide polymorphism, Chlamydia trachomatis, Biology, medicine.disease_cause, Microbiology, Genome, Article, 03 medical and health sciences, Plasmid, Virology, plasmid, Genetic variation, evolution, medicine, diagnostics, SNP, Gene, lcsh:QH301-705.5, 030304 developmental biology, Genetics, 0303 health sciences, Chlamydia, 030306 microbiology, sequencing, medicine.disease, lcsh:Biology (General), genetic variation
Abstract: Chlamydia trachomatis is an obligate intracellular pathogen of humans, causing both the sexually transmitted infection, chlamydia, and the most common cause of infectious blindness, trachoma. The majority of sequenced C. trachomatis clinical isolates carry a 7.5-Kb plasmid, and it is becoming increasingly evident that this is a key determinant of pathogenicity. The discovery of the Swedish New Variant and the more recent Finnish variant highlight the importance of understanding the natural extent of variation in the plasmid. In this study we analysed 524 plasmid sequences from publicly available whole-genome sequence data. Single nucleotide polymorphisms (SNP) in each of the eight coding sequences (CDS) were identified and analysed. There were 224 base positions out of a total 7550 bp that carried a SNP, which equates to a SNP rate of 2.97%, nearly three times what was previously calculated. After normalising for CDS size, CDS8 had the highest SNP rate at 3.97% (i.e., number of SNPs per total number of nucleotides), whilst CDS6 had the lowest at 1.94%. CDS5 had the highest total number of SNPs across the 524 sequences analysed (2267 SNPs), whereas CDS6 had the least SNPs with only 85 SNPs. Calculation of the genetic distances identified CDS6 as the least variable gene at the nucleotide level (d = 0.001), and CDS5 as the most variable (d = 0.007), however, at the amino acid level CDS2 was the least variable (d = 0.001), whilst CDS5 remained the most variable (d = 0.013). This study describes the largest in-depth analysis of the C. trachomatis plasmid to date, through the analysis of plasmid sequence data mined from whole genome sequences spanning 50 years and from a worldwide distribution, providing insights into the nature and extent of existing variation within the plasmid as well as guidance for the design of future diagnostic assays. This is crucial at a time when single-target diagnostic assays are failing to detect natural mutants, putting those infected at risk of a serious long-term and life-changing illness.
Published: 2020
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