Transmitochondrial embryonic stem cells containing pathogenic mtDNA mutations are compromised in neuronal differentiation

Objectives:  Defects of the mitochondrial genome (mtDNA) cause a series of rare, mainly neurological disorders. In addition, they have been implicated in more common forms of movement disorders, dementia and the ageing process. In order to try to model neuronal dysfunction associated with mitochondrial disease, we have attempted to establish a series of transmitochondrial mouse embryonic stem cells harbouring pathogenic mtDNA mutations. Materials and methods: Transmitochondrial embryonic stem cell cybrids were generated by fusion of cytoplasts carrying a variety of mtDNA mutations, into embryonic stem cells that had been pretreated with rhodamine 6G, to prevent transmission of endogenous mtDNA. Cybrids were differentiated into neurons and assessed for efficiency of differentiation and electrophysiological function. Results:  Neuronal differentiation could occur, as indicated by expression of neuronal markers. Differentiation was impaired in embryonic stem cells carrying mtDNA mutations that caused severe biochemical deficiency. Electrophysiological tests showed evidence of synaptic activity in differentiated neurons carrying non-pathogenic mtDNA mutations or in those that caused a mild defect of respiratory activity. Again, however, neurons carrying mtDNA mutations that resulted in severe biochemical deficiency had marked reduction in post-synaptic events. Conclusions:  Differentiated neurons carrying severely pathogenic mtDNA defects can provide a useful model for understanding how such mutations can cause neuronal dysfunction

Risk factors and variations in detection of new bovine tuberculosis breakdowns via slaughterhouse surveillance in Great Britain.

Slaughterhouse surveillance through post-mortem meat inspection provides an important mechanism for detecting bovine tuberculosis (bTB) infections in cattle herds in Great Britain (GB), complementary to the live animal skin test based programme. We explore patterns in the numbers of herd breakdowns detected through slaughterhouse surveillance and develop a Bayesian hierarchical regression model to assess the associations of animal-level factors with the odds of an infected animal being detected in the slaughterhouse, allowing us to highlight slaughterhouses that show atypical patterns of detection. The analyses demonstrate that the numbers and proportions of breakdowns detected in slaughterhouses increased in GB over the period of study (1998-2013). The odds of an animal being a slaughterhouse case was strongly associated with the region of the country that the animal spent most of its life, with animals living in high-frequency testing areas of England having on average 21 times the odds of detection compared to animals living in Scotland. There was also a strong effect of age, with animals slaughtered at > 60 months of age having 5.3 times the odds of detection compared to animals slaughtered between 0-18 months of age. Smaller effects were observed for cattle having spent time on farms with a history of bTB, quarter of the year that the animal was slaughtered, movement and test history. Over-and-above these risks, the odds of detection increased by a factor of 1.1 for each year of the study. After adjustment for these variables, there were additional variations in risk between slaughterhouses and breed. Our framework has been adopted into the routine annual surveillance reporting carried out by the Animal Plant Health Agency and may be used to target more detailed investigation of meat inspection practices.Defra Project SE3133. (Department of Environment and Rural Affairs, UK Government

Synergistic Positive Feedback Mechanisms Underlying Seizure Initiation

Investigations into seizure initiation, in recent years, have focused almost entirely upon alterations of interneuronal function, chloride homeostasis, and extracellular potassium levels. In contrast, little attention has been directed toward a possible role of dendritic plateau potentials in the actual ictogenic transition, despite a substantial literature dating back 40 years regarding its importance generally in epilepsy. Here, we argue that an increase in dendritic excitability, coordinated across the population of pyramidal cells, is a key stage in ictogenesis

Efficient Bayesian model choice for partially observed processes: with application to an experimental transmission study of an infectious disease

Infectious diseases such as avian influenza pose a global threat to human health. Mathematical and statistical models can provide key insights into the mechanisms that underlie the spread and persistence of infectious diseases, though their utility is linked to the ability to adequately calibrate these models to observed data. Performing robust inference for these systems is challenging. The fact that the underlying models exhibit complex non-linear dynamics, coupled with practical constraints to observing key epidemiological events such as transmission, requires the use of inference techniques that are able to numerically integrate over multiple hidden states and/or infer missing information. Simulation-based inference techniques such as Approximate Bayesian Computation (ABC) have shown great promise in this area, since they rely on the development of suitable simulation models, which are often easier to code and generalise than routines that require evaluations of an intractable likelihood function. In this manuscript we make some contributions towards improving the efficiency of ABC-based particle Markov chain Monte Carlo methods, and show the utility of these approaches for performing both model inference and model comparison in a Bayesian framework. We illustrate these approaches on both simulated data, as well as real data from an experimental transmission study of highly pathogenic avian influenza in genetically modi fied chickens.This work was supported by the Biotechnology and Biological Sciences Research Council (grants BB/G00479X/1, BBS/B/00239, and BBS/B/00301) and by the Cambridge Infectious Diseases Consortium Department for Environment, Food and Rural Affairs-Higher Education Funding Council for England (grant VT0105). AJKC was supported by BBSRC grant BB/I024550/1

The demography of free-roaming dog populations and applications to disease and population control

Understanding the demography of domestic dog populations is essential for effective disease control, particularly of canine-mediated rabies. Demographic data are also needed to plan effective population management. However, no study has comprehensively evaluated the contribution of demographic processes (i.e. births, deaths and movement) to variations in dog population size or density, or determined the factors that regulate these processes, including human factors. We report the results of a 3-year cohort study of domestic dogs, which is the first to generate detailed data on the temporal variation of these demographic characteristics. The study was undertaken in two communities in each of Bali, Indonesia and Johannesburg, South Africa, in rabies-endemic areas and where the majority of dogs were free-roaming. None of the four communities had been engaged in any dog population management interventions by local authorities or animal welfare organizations. All identified dogs in the four communities were monitored individually throughout the study. We observed either no population growth or a progressive decline in population size during the study period. There was no clear evidence that population size was regulated through environmental resource constraints. Rather, almost all of the identified dogs were owned and fed regularly by their owners, consistent with population size regulated by human demand. Finally, a substantial fraction of the dogs originated from outside the population, entirely through the translocation of dogs by people, rather than from local births. These findings demonstrate that previously reported growth of dog populations is not a general phenomenon and challenge the widely held view that free-roaming dogs are unowned and form closed populations. Synthesis and applications. These observations have broad implications for disease and population control. The accessibility of dogs for vaccination and evaluation through owners and the movement of dogs (some of them infected) by people will determine the viable options for disease control strategies. The impact of human factors on population dynamics will also influence the feasibility of annual vaccination campaigns to control rabies and population control through culling or sterilization. The complex relationship between dogs and people is critically important in the transmission and control of canine-mediated rabies. For effective management, human factors must be considered in the development of disease and population control programmes

Age-dependent patterns of bovine tuberculosis in cattle.

Bovine tuberculosis (BTB) is an important livestock disease, seriously impacting cattle industries in both industrialised and pre-industrialised countries. Like TB in other mammals, infection is life long and, if undiagnosed, may progress to disease years after exposure. The risk of disease in humans is highly age-dependent, however in cattle, age-dependent risks have yet to be quantified, largely due to insufficient data and limited diagnostics. Here, we estimate age-specific reactor rates in Great Britain by combining herd-level testing data with spatial movement data from the Cattle Tracing System (CTS). Using a catalytic model, we find strong age dependencies in infection risk and that the probability of detecting infection increases with age. Between 2004 and 2009, infection incidence in cattle fluctuated around 1%. Age-specific incidence increased monotonically until 24-36 months, with cattle aged between 12 and 36 months experiencing the highest rates of infection. Beef and dairy cattle under 24 months experienced similar infection risks, however major differences occurred in older ages. The average reproductive number in cattle was greater than 1 for the years 2004-2009. These methods reveal a consistent pattern of BTB rates with age, across different population structures and testing patterns. The results provide practical insights into BTB epidemiology and control, suggesting that targeting a mass control programme at cattle between 12 and 36 months could be beneficial.EBP is funded by an Engineering and Physical Sciences Research Council (EPSRC) fellowship. JLNW is supported by the Alborada Trust and the RAPIDD program of the Science & Technology Directorate, U.S. Department of Homeland Security, and the Fogarty International Center, U.S. National Institutes of Health. AJKC is supported by Defra grant no. SE-3127. TJM is supported by the BBSRC. We thank Steve Holdship and Rose Nicholson at Defra the AHVLA team for providing access to the CTS and VetNet

Two common psychophysical measures of surround suppression reflect independent neuronal mechanisms.

Psychophysical surround suppression is believed to reflect inhibitory neuronal mechanisms in visual cortex. In recent years, two psychophysical measures of surround suppression have been much studied: (i) duration thresholds on a motion-discrimination task (which are worse for larger than for smaller stimuli) and (ii) contrast thresholds on a contrast-detection task (which are worse when grating stimuli are surrounded by a stimulus of the same orientation than when they are presented in isolation or surrounded by a stimulus of orthogonal orientation). Changes in both metrics have been linked to several different human conditions, including aging, differences in intelligence, and clinical disorders such as schizophrenia, depression, and autism. However, the exact nature of the neuronal correlate underlying these phenomena remains unclear. Here, we use an individual-differences approach to test the hypothesis that both measures reflect the same property of the visual system, e.g., the strength of GABA-ergic inhibition across visual cortex. Under this hypothesis we would expect the two measures to be significantly positively correlated across individuals. In fact, they are not significantly correlated. In addition, we replicate the previously reported correlation between age and motion-discrimination surround suppression, but find no correlation between age and contrast-detection surround suppression. We conclude that the two forms of psychophysical surround suppression arise independently from different cortical mechanisms

Mechanisms underlying different onset patterns of focal seizures

Focal seizures are episodes of pathological brain activity that appear to arise from a localised area of the brain. The onset patterns of focal seizure activity have been studied intensively, and they have largely been distinguished into two types - low amplitude fast oscillations (LAF), or high amplitude spikes (HAS). Here we explore whether these two patterns arise from fundamentally different mechanisms. Here, we use a previously established computational model of neocortical tissue, and validate it as an adequate model using clinical recordings of focal seizures. We then reproduce the two onset patterns in their most defining properties and investigate the possible mechanisms underlying the different focal seizure onset patterns in the model. We show that the two patterns are associated with different mechanisms at the spatial scale of a single ECoG electrode. The LAF onset is initiated by independent patches of localised activity, which slowly invade the surrounding tissue and coalesce over time. In contrast, the HAS onset is a global, systemic transition to a coexisting seizure state triggered by a local event. We find that such a global transition is enabled by an increase in the excitability of the "healthy" surrounding tissue, which by itself does not generate seizures, but can support seizure activity when incited. In our simulations, the difference in surrounding tissue excitability also offers a simple explanation of the clinically reported difference in surgical outcomes. Finally, we demonstrate in the model how changes in tissue excitability could be elucidated, in principle, using active stimulation. Taken together, our modelling results suggest that the excitability of the tissue surrounding the seizure core may play a determining role in the seizure onset pattern, as well as in the surgical outcome

Potential Benefits of Cattle Vaccination as a Supplementary Control for Bovine Tuberculosis

This is the final published version of the article. It was originally published in PLOS Computational Biology (Conlan AJK, Brooks Pollock E, McKinley TJ, Mitchell AP, Jones GJ, Vordermeier M, Wood JLN, PLoS Computational Biology 2015, 11(2): e1004038. doi:10.1371/journal.pcbi.1004038).\ud \ud ? 2015 Crown Copyright. This is an open-access article distributed under the terms of the free Open Government Licence, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited. See: http://www.nationalarchives.gov.uk/doc/o?pen-government-licence/open-government-l?icence.htmVaccination for the control of bovine tuberculosis (bTB) in cattle is not currently used within any international control program, and is illegal within the EU. Candidate vaccines, based upon Mycobacterium bovis bacillus Calmette-Gu?rin (BCG) all interfere with the action of the tuberculin skin test, which is used to determine if animals, herds and countries are officially bTB-free. New diagnostic tests that Differentiate Infected from Vaccinated Animals (DIVA) offer the potential to introduce vaccination within existing eradication programs. We use within-herd transmission models estimated from historical data from Great Britain (GB) to explore the feasibility of such supplemental use of vaccination. The economic impact of bovine Tuberculosis for farmers is dominated by the costs associated with testing, and associated restrictions on animal movements. Farmers? willingness to adopt vaccination will require vaccination to not only reduce the burden of infection, but also the risk of restrictions being imposed. We find that, under the intensive sequence of testing in GB, it is the specificity of the DIVA test, rather than the sensitivity, that is the greatest barrier to see a herd level benefit of vaccination. The potential negative effects of vaccination could be mitigated through relaxation of testing. However, this could potentially increase the hidden burden of infection within Officially TB Free herds. Using our models, we explore the range of the DIVA test characteristics necessary to see a protective herd level benefit of vaccination. We estimate that a DIVA specificity of at least 99.85% and sensitivity of >40% is required to see a protective benefit of vaccination with no increase in the risk of missed infection. Data from experimentally infected animals suggest that this target specificity could be achieved in vaccinates using a cocktail of three DIVA antigens while maintaining a sensitivity of 73.3% (95%CI: 61.9, 82.9%) relative to post-mortem detection.This study was funded by Defra project SE3127 and uses nationally collected incidence and cattle-movement data sets held by Defra. The funders had no role in study design, data analysis, decision to publish, or preparation of the manuscript

Potential benefits of cattle vaccination as a supplementary control for bovine tuberculosis

Published onlineJournal ArticleResearch Support, Non-U.S. Gov'tVaccination for the control of bovine tuberculosis (bTB) in cattle is not currently used within any international control program, and is illegal within the EU. Candidate vaccines, based upon Mycobacterium bovis bacillus Calmette-Guérin (BCG) all interfere with the action of the tuberculin skin test, which is used to determine if animals, herds and countries are officially bTB-free. New diagnostic tests that Differentiate Infected from Vaccinated Animals (DIVA) offer the potential to introduce vaccination within existing eradication programs. We use within-herd transmission models estimated from historical data from Great Britain (GB) to explore the feasibility of such supplemental use of vaccination. The economic impact of bovine Tuberculosis for farmers is dominated by the costs associated with testing, and associated restrictions on animal movements. Farmers' willingness to adopt vaccination will require vaccination to not only reduce the burden of infection, but also the risk of restrictions being imposed. We find that, under the intensive sequence of testing in GB, it is the specificity of the DIVA test, rather than the sensitivity, that is the greatest barrier to see a herd level benefit of vaccination. The potential negative effects of vaccination could be mitigated through relaxation of testing. However, this could potentially increase the hidden burden of infection within Officially TB Free herds. Using our models, we explore the range of the DIVA test characteristics necessary to see a protective herd level benefit of vaccination. We estimate that a DIVA specificity of at least 99.85% and sensitivity of >40% is required to see a protective benefit of vaccination with no increase in the risk of missed infection. Data from experimentally infected animals suggest that this target specificity could be achieved in vaccinates using a cocktail of three DIVA antigens while maintaining a sensitivity of 73.3% (95%CI: 61.9, 82.9%) relative to post-mortem detection.This study was funded by Defra project SE3127 and uses nationally collected incidence and cattle-movement data sets held by Defra