Election to St. John's College

I'm pleased to have been elected to membership of the SCR at St. John's College, my alma mater. The college is an important aspect of life at Oxford as it gives an alternative centre of gravity outside the department for participation in social and academic activities. As a member of University staff with solely research responsibilities, it is a welcome opportunity to interact with the community of teaching fellows, students and junior researchers who belong to the college.

The picture is of the Spring crocus lawn taken in the college gardens.

Postdoc and PhD position available

These positions are now closed.

Advertised today in Nature and on Thursday in New Scientist are two positions in my lab. I am looking for a postdoc and a PhD student to work on the genome evolution and epidemiology of four human pathogens as part of the Modernising Medical Microbiology project. Three of the pathogens share the theme of hospital-acquired infections: they are Staphylococcus aureus (of MRSA infamy), Clostridium difficile and norovirus (aka winter vomiting disease). The fourth is Mycobacterium tuberculosis (TB) which is a re-emerging problem in developed countries.

The aim of the project is to use whole genome sequencing of many isolates (100s to 1000s) in order to reconstruct evolutionary relationships and deconstruct transmission routes. We hope to develop the technology to the stage that we can trace the spread of pathogens in real time, and uncover the epidemiological triggers for the spread of disease.

As of January I have relocated to the Nuffield Department of Clinical Medicine at the University of Oxford, and the project is a collaborative affair between people at Oxford (including Rory Bowden, Derrick Crook, Peter Donnelly and Rosalind Harding), the Wellcome Trust Sanger Institute, the NHS and the Health Protection Agency. The project is funded by the UKCRC and further details of the positions are available online for the postdoc and PhD studentship. The closing date for applications is Friday, 2 April 2010.

Holding early human stone tools

Today I had an extraordinary experience, precipitated by my visit to the British Museum on something of a whim. Listening to the Radio 4 series A History of the World in 100 Objects, my imagination had been captured by the descriptions of early stone tools - a chopper and a hand axe - featured in the first couple of programmes in the series. These tools, which were found in the Olduvai Gorge, in modern-day Tanzania, are examples of the oldest known objects made by humans. What is fascinating is that their simple design belies a capacity for mental forethought. They are tangible evidence that the humans living 2 million years ago had the intelligence to conceive of and the dexterity to manufacture tools.
I had been visiting friends in London, and before leaving I decided to pass by the museum to see these relics for myself. I found the stone tools in a dim room in the near corner of the museum, shielded by glass cases. After reading the descriptions and wandering round I noticed a lady showing some children a bunch of similar-looking objects she had in a wooden box. I asked if they were casts and could hardly believe it when she told me it was the real thing. Two stone hand axes, 1 million years old, made from basalt and quartz, and a basalt chopper, 2 million years old - the oldest items in the museum. To hold in the palm of my hand a tool fashioned 2 million years ago by a cognizant proto-human, I could imagine the heavy object fitting just as neatly into the hand of its designer, and in trying to understand the way it might have been used to butcher carcasses, pound meat and scrape flesh off bones I felt I got a brief glimpse into the intentions of its designer. The study of evolution rarely affords such vivid connections with its subject matter, and I felt privileged to stumble across such an encounter today.

Campylobacter source attribution in New Zealand

What is the source of the common food poisoning pathogen Campylobacter jejuni was the subject of a paper published in September last year in PLoS Genetics by my colleagues and I, in which we traced the origin of bacterial isolates collected from patients in Lancashire, England. In that study, and a subsequent investigation into campylobacteriosis across Scotland, we found that the majority of cases could be attributed to populations of C. jejuni typically found in poultry.

Now Petra Mullner, Nigel French and colleagues have genetically characterized the C. jejuni populations found in human patients, cattle, sheep, poultry and environmental samples from New Zealand covering the period March 2005 - February 2008. What is special about their study is that the New Zealand poultry industry is a closed system, with no foreign imports, making it possible to directly sample the putative source populations and disease-causing isolates concurrently.

Like the studies in England and Scotland, poultry was the inferred source of the majority of disease in New Zealand. Uniquely however, it was possible to attribute cases separately to the three major poultry suppliers on the islands. One supplier in particular was attributed a disproportionate number of cases using 3 assignment methods, including my method (iSource, soon to be available on this website). Supported in part by this evidence, the New Zealand Food Safety Authority introduced mandatory targets for limiting Campylobacter contamination of poultry products in 2007. Remarkably, the number of cases fell from 15,873 in 2006 before the control measures were introduced to 6,689 in 2008. The next chapter of this intriguing story will be a follow-up study to establish whether the fall in the number of cases corresponded to a reduction in the proportion of campylobacteriosis attributable to poultry sources.

Selection in a putative meningitis vaccine target

In Variation of the factor H-binding protein in Neisseria meningitidis, Carina Brehony in Martin Maiden's lab at Oxford investigated a group of outer membrane proteins in the bacterium responsible for meningococcal meningitis. To date, attempts to raise a vaccine against the common serogroup B meningococci have been frustrated by the low immunogenicity of the serogroup B capsular polysaccharide, despite success with serogroups A and C. Outer membrane proteins, such as factor H-binding protein (fHbp) may provide alternative targets for vaccine development.

However, fHbp is genetically diverse, and our investigation showed evidence of structuring into three groups. OmegaMap analyses of the three groups revealed a signature consistent with strong selection pressure for antigenic variability at the gene. Notably, there was clear evidence of diversifying selection at several previously discovered epitopes - positions in the protein targeted by antibodies during bacteria-killing immune response. (Analysis of one group is shown in the figure, with known epitopes marked).

While these observations are encouraging in terms of understanding the biology of pathogen antigens, a pressing question is how do we translate that understanding into practical vaccine design? Studies such as ours suggest a multi-component vaccine may be necessary to achieve broad coverage against serogroup B meningococci.

Recombination and proper segregation in human meiosis

My blog entries have lapsed since the summer while I have attempted to press on with various projects to tie up as much as possible by the end of the year. Meanwhile, my collaborators and I have had three papers published.

In Broad-scale recombination patterns underlying proper disjunction in humans, Adi Alon and colleagues have used a large Hutterite pedigree to test two molecular hypotheses in a statistical genetics fashion. Crossing-over is important for proper segregation of chromosomes during meiosis. When chromosomes fail to segregate properly, the result is aneuploidy, a genetic pathology underlying many inherited diseases; for example, aneuploidy at chromosome 21 is often the basis of Down's syndrome.


It has been suggested that a hard limit of at least one crossover per chromosome is necessary for correct disjunction; others have suggested the requirement is for one crossover per chromosome arm. By reconstructing the probable distribution of the number of crossovers during meiosis, we were able to show that proper disjunction frequently occurs in humans in the absence of a crossover every chromosome arm. Further, the evidence suggested that successful segregation of some chromosomes can occur without a crossover at all - interestingly chromosome 21 was flagged up among others. This leads to the question, is there a back-up cellular mechanism to rescue meiotic division when crossovers fail to form?

SMBE Iowa City

I spent the beginning of the month at the SMBE (Society for Molecular Biology and Evolution) conference in Iowa City. It was a good chance to catch up with people and find out what research is going on in the field, as well as to speak with collaborators about on-going projects. One of those is Peter Andolfatto, who works on genome evolution in Drosophila species. Molly and I are collaborating with Peter on a project to detect natural selection within and between Drosophila species. The main idea is to improve inference by taking into account variation in selection pressure throughout the gene. Our method draws on the advantages of a number of current approaches such as Rasmus Nielsen and Ziheng Yang's codeml package (part of PAML), Carlos Bustamante's MKPRF (McDonald-Kreitman Poisson Random Field) model and Gil McVean and my program omegaMap in that it exploits patterns of polymorphism within and between species, while allowing for conservation and adaptation within the same gene. You can view the slides of my SMBE talk here, which was titled "Adaptive events in hominid (and Drosophila) evolution".

Science Bomb!

On Friday Chris Spencer gave the PPS (Pritchard/Przeworski/Stephens) lab meeting as part of a trip to Chicago. Chris talked about his work in Oxford on association studies in a number of common genetic diseases being studied by the Wellcome Trust Case Control Consortium.

Beforehand I dropped the Science Bomb, a new innovation this year (for which I think Barbara Engelhardt is responsible) where someone talks about a particularly interesting or timely article. Dan Gaffney pointed me in the direction of a PLoS Biology paper titled Reawakening Retrocyclins: Ancestral Human Defensins Active Against HIV-1.

The subject of the study is a human pseudogene known as retrocyclin, which has been shown to confer resistance to HIV-1 infection in human cell lines. The pseudogene is expressed naturally in several human tissues, but not translated into protein owing to a premature stop codon. The paper's authors reawakened retrocyclin using aminoglycosides, a class of antibiotics that cause (as a side effect) a degree of mis-translation and hence allow "read-through" of the stop codon. You can see the slides from my Science Bomb here.

Neolithic origin of Campylobacter jejuni

As part of a recent trip to the University of Edinburgh to visit Andrew Rambaut, I gave a talk on some work of mine published in the February edition of Molecular Biology and Evolution and subsequently recommended on the Faculty of 1000 website about the evolution of the gut pathogen Campylobacter jejuni.

Part of the paper is concerned with the issue of the timescale of Campylobacter evolution, and using longitudinal samples of C. jejuni DNA sequences we attempted to calibrate the molecular clock in a similar way to that which is standard practice for viruses.

We detected surprisingly rapid evolution - 1,000 times faster than traditional estimates - which would place the split of C. jejuni from its closest relative C. coli during the Neolithic revolution. Interestingly, the point estimate of 6,500 years ago for the split from C. coli - which preferentially infects swine - coincides with the spread of pig domestication in the Near East and Europe in the 4th millennium BC.

The date is controversial because the traditional dating method, which is based on bounding deep phylogenetic splits such as the common ancestor of mitochondria and bacteria, would place the divergence of C. jejuni and C. coli closer to 10 million years ago.

After the seminar I had an interesting discussion with Paul Sharp, who was in the audience. Prof Sharp is actively researching the causes of conflict between long-term and short-term estimates of the rate of evolution in viruses. As he points out, short-term rate estimates (usually based on longitudinally-sampled viral sequences) frequently suggest that evolution is occurring much more rapidly than long-term estimates (based on deeper calibration points, such as co-phylogeny of host and pathogen). This phenomenon, observed in HIV and hepatitis C among others, may be caused by overly simplistic models of sequence evolution.

So how plausible is it that a ubiquitous bacterial pathogen such as C. jejuni evolved as recently as the Neolithic, possibly in response to changes brought about by agriculture or animal husbandry? Longitudinal studies of Helicobacter pylori and Neisseria gonnorhoeae have obtained similarly rapid rates of bacterial evolution, and evidence is mounting that the Neolithic revolution played an important role in creating new niches for human, plant and animal pathogens. Perhaps the best prospect for resolving these questions will be studies of ancient DNA preserved from the period in question.

omegaMap at BioHPC

All evolutionary biologists wishing to make use of omegaMap now have access to a high performance parallel computing cluster via the internet courtesy of Cornell's CBSU and Microsoft. The software, which allows the detection of selection and recombination in DNA or RNA sequences, can be run via the web interface at cbsuapps.tc.cornell.edu/omegamap.aspx, or downloaded as part of the BioHPC suite.

The web interface consists of a simple form where users can upload their configuration file and sequences in FASTA format. Completed jobs are notified by e-mail. To learn more about the project visit the CBSU home page.

Meanwhile, I am working on several major updates to omegaMap, the most interesting of which will probably be the development of a new model that allows for the joint analysis of natural selection acting on sequences from different populations or species. The aim is to integrate population genetic and phylogenetic models of selection in order to exploit the signal of selection contained both in polymorphism within populations (or species) and divergence between them. I will be presenting progress on this work, in the context of hominid evolution, at the 2009 SMBE meeting in Iowa City this June.

Human Evolution in New York City

Rounding off a hectic end to 2008 was a trip to visit Molly, currently on sabbatical in New York city. Joanna and I flew out to spend the final weekend before Christmas discussing projects and frequenting the local coffee shops, restaurants and bars. I took the opportunity to visit the American Museum of Natural History adjacent to Central Park after reading about its dinosaur collections in the Catcher in the Rye; pictured is an Allosaurus skeleton, which stands in the main entrance hall. Of particular interest was the Spitzer Hall of Human Origins which features a wealth of fossil remains and artefacts including a cast of the Laetoli footprints and a diorama of an Australopithecus afarensis nuclear family. Fittingly, the very focus of the New York trip was to discuss the on-going project to characterize natural selection between hominid species.

Inferring niche membership from genetic diversity

Each Wednesday the Ecology and Evolution department run a journal club called Noon Illumination, and this week I volunteered to lead discussion on a recent article titled Resource Partitioning and Sympatric Differentiation Among Closely Related Bacterioplankton (Science 320: 1081-5), by Dana Hunt and colleagues based at MIT and Ghent. I originally prepared the presentation for a Bacterial Metagenomics workshop in Berlin this July, organized by Daniel Falush.

Of central interest in the paper is a novel methodology that infers habitat/niche based on ecological variables and DNA sequencing in the family of marine bacteria Vibrionaceae. That places it in the wider context of methods that attempt to predict phenotype (in this case niche) from genotype. Their approach is an elegant extension of familiar phylogenetic methods to model habitat switching over evolutionary time. Based on arguments put forward by Christophe Fraser and colleagues, the paper reasons that the ancestral habitat switches they detect are likely to be adaptive because the rate of recombination eclipses the mutation rate sufficiently to preclude the possibility of neutral genetic clustering.

However the high rate of recombination raises some difficulties of interpretation. The principal phylogenetic reconstruction was based on the hsp60 gene, but by sequencing other housekeeping genes, Hunt and colleagues found that in some cases, recombination between genes caused an artefactual habitat switch in the hsp60 ancestry that was not evident in the other genes. Using a permutation test, I found evidence for recombination within the vibrio hsp60 genes, which may confound the phylogenetic reconstruction of evolutionary relationships (Schierup and Hein 2000). On a more philosophical note, suppose you could directly observe ancestral habitat switches. Would that be strong evidence for adaptation? An association between habitat and genetic lineage is probably not sufficient to demonstrate the action of natural selection. On the other hand, frequent recombination could empower genome-wide scans for extreme association between genes and habitats, that would provide stronger support for adaptation.

You can view a PDF of the presentation of this stimulating article in our journal club here.

Tracing the source of campylobacteriosis

Finally, it's out! The main piece of work to come out of my two-year period as Research Associate at Lancaster University is published today in PLoS Genetics.

The article reports a study in Lancashire, England, of the bacterium Campylobacter jejuni, the primary cause of bacterial gastro-enteritis in developed countries. We inferred the source of infection in 1,200 patients by comparing the DNA sequences of C. jejuni taken from those patients to 1,100 taken from different animal species and the environment. The result: livestock are the source of infection in 97% of cases.

In addition to preparing the figures, approving final drafts, and producing a press release in conjunction with the PLoS Genetics and university press offices, I have spent much of my time over the last three weeks revising a companion paper on the evolution of C. jejuni. On Friday that was resubmitted to Molecular Biology and Evolution, and should it be accepted, will draw a line under my Lancaster projects.

Visit to Kilifi

The final days in Paris were taken up by revisions to a paper submitted in April to PLoS Genetics. With luck the revisions will be accepted and that paper will be coming out soon. From Paris I flew to Mombasa, Kenya to begin a 3 week collaboration with Caroline Buckee at the KEMRI-Wellcome Trust research unit in Kilifi. Caroline, Pete Bull and others at Kilifi work on the evolution of var genes in Plasmodium falciparum, the most common and lethal agent of malaria. The var genes encode a family of proteins expressed by the pathogen on the membrane of infected red blood cells. Implicated in pathogenesis, these genes are highly diverse in order to evade the host immune system. The first step in piecing together their evolutionary history is to align the sequences - a task made difficult by the abundance of insertions and deletions. From there we hope to characterize the relative importance of gene duplication and homologous and non-homologous recombination in driving the evolution of these genes.

Exchange with the Institut Pasteur

My position at the University of Chicago is funded by a grant awarded to Molly by the National Institutes of Health (NIH) to detect the signature that natural selection has left on the human genome. One of our collaborations is with the human genetics lab of Lluis Quintana-Murci at the Institut Pasteur in Paris, and I'm making the first of a number of exchange visits to strengthen the ties between two labs.

Lluis' group are interested in the selection pressure that pathogens have exerted on the human genome, and in particular a family of genes involved in the immune system known as toll-like receptors. The idea is that together we develop methods to detect and quantify selection in these genes by comparison to neutral regions in individuals from around the world. Among the people involved in the project is Luis Barreiro, a post-doc who has just arrived in the Human Genetics department at Chicago from the Institut Pasteur.

In Paris I've been participating in group meetings, offering my opinion on manuscripts coming out of Lluis' lab, and discussing with lab members how we might analyze the DNA sequence data they're producing. I've also been acquainting myself with the produce of Château des Ravatys (the Institut Pasteur vinyard) and celebrating the 14 Juillet (photo).

What do researchers do?

Most of my friends (and maybe even my colleagues) don't have much idea of what I do day-to-day. That's the subject of this blog. Modern research is quantified in terms of published articles, but that forms just a small part of daily working life, and the process from conceiving an interesting question or idea to published article is a long and tortuous one. Often articles aren't published until months or years after the research was conducted, and so there is a time-lag between what appears in print and what the current focus of research is. By that time the initial excitement of pursuing novel work is a distant memory, having been supplanted by several rounds of peer review, revision and resubmission.

So this is an attempt to communicate what I am doing now, to talk about what I think's exciting this week (or this afternoon) and to show what being a researcher means on a daily basis.