Lack of RNA-DNA oligonucleotide (chimeraplast) mutagenic activity in mouse embryos

There are numerous reports of the use of RNA-DNA oligonucleoticles (chimeraplasts) to correct point mutations in vitro and in vivo, including the human apolipoprotein E gene (ApoE). Despite the absence of selection for targeting, high efficiency conversion has been reported. Although mainly used to revert deleterious mutations for gene therapy applications, successful use of this approach would have the potential to greatly facilitate the production of defined mutations in mice and other species. We have attempted to create a point mutation in the mouse ApoE gene by microinjection of chimeraplast into the pronuclei of 1-cell mouse eggs. Following transfer of microinjected eggs we analysed 139 E12.5 embryos, but obtained no evidence for successful conversion. (c) 2005 Wiley-Liss, Inc

Correction of the neuropathogenic human apolipoprotein E4 (APOE4) gene to APOE3 in vitro using synthetic RNA/DNA oligonucleotides (chimeraplasts)

Apolipoprotein E (apoE) is a multifunctional circulating 34-kDa protein, whose gene encodes single-nucleotide polymorphisms linked to several neurodegenerative diseases. Here, we evaluate whether synthetic RNA/DNA oligonucleoticles (chimeraplasts) can convert a dysfunctional gene, APOE4 (C -> T, Cys112Arg), a risk factor for Alzheimer's disease and other neurological disorders, into wild-type APOE3. In preliminary experiments, we treated recombinant Chinese hamster ovary (CHO) cells stably secreting apoE4 and lymphocytes from a patient homozygous for the epsilon 4 allele with a 68-mer apoE4-to-apoE3 chimeraplast, complexed to the cationic delivery reagent, polyethyleneimine. Genotypes were analyzed after 48 h by routine polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and by genomic sequencing. Clear conversions of APOE4 to APOE3 were detected using either technique, although high concentrations of chimeraplast were needed (>= 800 nM). Spiking experiments of PCR reactions or CHO-K1 cells with the chimeraplast confirmed that the repair was not artifactual. However, when treated recombinant CHO cells were passaged for 10 d and then subcloned, no conversion could be detected when > 90 clones were analyzed by locus-specific PCR-RFLP. We conclude that the apparent efficient repair of the APOE4 gene in CHO cells or lymphocytes 48 h post-treatment is unstable, possibly because the high levels of chimeraplast and polyethyleneimine that were needed to induce nucleotide substitution are cytotoxic

Comment on "Wandering minds: The default network and stimulus-independent thought"

Mason et al. (Reports, 19 January 2007, p. 393) attributed activity in certain regions of the "resting" brain to the occurrence of mind-wandering. However, previous research has demonstrated the difficulty of distinguishing this type of stimulus-independent thought from stimulus-oriented thought (e.g., watchfulness). Consideration of both possibilities is required to resolve this ambiguity

Distinct neural mechanisms underlie the success, precision, and vividness of episodic memory

A network of brain regions have been linked with episodic memory retrieval, but limited progress has been made in identifying the contributions of distinct parts of the network. Here, we utilized continuous measures of retrieval to dissociate three components of episodic memory: retrieval success, precision, and vividness. In the fMRI scanner, participants encoded objects that varied continuously on three features: color, orientation, and location. Participants' memory was tested by having them recreate the appearance of the object features using a continuous dial, and continuous vividness judgments were recorded. Retrieval success, precision, and vividness were dissociable both behaviorally and neurally: successful versus unsuccessful retrieval was associated with hippocampal activity, retrieval precision scaled with activity in the angular gyrus, and vividness judgments tracked activity in the precuneus. The ability to dissociate these components of episodic memory reveals the benefit afforded by measuring memory on a continuous scale, allowing functional parcellation of the retrieval network.James S McDonnell Foundation Scholar Award, Medical Research Council, Wellcome Trust, Economic and Social Research Counci

Brain Mechanisms of Reality Monitoring

Reality monitoring processes are necessary for discriminating between internally generated information and information that originated in the outside world. They help us to identify our thoughts, feelings, and imaginations, and to distinguish them from events we may have experienced or have been told about by someone else. Reality monitoring errors range from confusions between real and imagined experiences, that are byproducts of normal cognition, to symptoms of mental illness such as hallucinations. Recent advances support an emerging neurocognitive characterization of reality monitoring that provides insights into its underlying operating principles and neural mechanisms, the differing ways in which impairment may occur in health and disease, and the potential for rehabilitation strategies to be devised that might help those who experience clinically significant reality monitoring disruption.Preparation of this article was supported by a James S. McDonnell Foundation Scholar Award to J.S.S., and by the University of Cambridge Behavioural and Clinical Neuroscience Institute, funded by a joint award from the Medical Research Council and the Wellcome Trust

Obesity and insulin resistance are associated with reduced activity in core memory regions of the brain

Increasing research in animals and humans suggests that obesity may be associated with learning and memory deficits, and in particular with reductions in episodic memory. Rodent models have implicated the hippocampus in obesity-related memory impairments, but the neural mechanisms underlying episodic memory deficits in obese humans remain undetermined. In the present study, lean and obese human participants were scanned using fMRI while completing a What-Where-When episodic memory test (the “Treasure-Hunt Task”) that assessed the ability to remember integrated item, spatial, and temporal details of previously encoded complex events. In lean participants, the Treasure-Hunt task elicited significant activity in regions of the brain known to be important for recollecting episodic memories, such as the hippocampus, angular gyrus, and dorsolateral prefrontal cortex. Both obesity and insulin resistance were associated with significantly reduced functional activity throughout the core recollection network. These findings indicate that obesity is associated with reduced functional activity in core brain areas supporting episodic memory and that insulin resistance may be a key player in this association.This study was funded by an MRC Centenary Early Career Award to LGC and a James S. McDonnell Foundation Scholar Award to JSS. LGC was funded by the Sarah Woodhead Research Fellowship at Girton College Cambridge. NSC was funded by an Experimental Psychology Society Mid-Career Award and the ERC

Distinct Roles for the Anterior Cingulate and Dorsolateral Prefrontal Cortices During Conflict Between Abstract Rules

Distinct patterns of activity within the anterior cingulate cortex (ACC) and dorsolateral prefrontal cortex (dlPFC) reported in neuroimaging studies during tasks involving conflict between competing responses have often been cited as evidence for their key contributions to conflict-monitoring and behavioral adaptation, respectively. However, supporting evidence from neuropsychological patients has been scarce and contradictory. We administered a well-studied analog of the Wisconsin Card Sorting Test, designed to elicit conflict between 2 abstract rules, to a cohort of 6 patients with damage to ACC or dlPFC. Patients who had sustained more significant damage to the ACC were not impaired either on a measure of “conflict cost” nor on measures of “conflict-induced behavioral adaptation.” In contrast, damage to dlPFC did not affect the conflict cost measure but abolished the patients’ ability to adapt their behavior following exposure to conflict, compared with controls. This pattern of results complements the findings from nonhuman primates with more circumscribed lesions to ACC or dlPFC on the same task and provides converging evidence that ACC is not necessary for performance when conflict is elicited between 2 abstract rules, whereas dlPFC plays a fundamental role in behavioral adaptation.The UK Medical Research Council

Eye movements reveal a dissociation between memory encoding and retrieval in adults with autism

People with Autism Spectrum Disorder (ASD) exhibit subtle deficits in recollection, which have been proposed to arise from encoding impairments, though a direct link has yet to be demonstrated. In the current study, we used eye-tracking to obtain trial-specific measures of encoding (eye movement patterns) during incidental (natural viewing) and intentional (strategic) encoding conditions in adults with ASD and typical controls. Using this approach, we tested the degree to which differences in encoding might contribute to recollection impairments, or whether group differences in memory primarily emerge at retrieval. Following encoding of scenes, participants were asked to distinguish between old and similar lure scenes and provide 'remember'/'familiar' responses. Intentional encoding increased eye movements and subsequent recollection in both groups to a similar degree, but the ASD group were impaired overall at the memory task and used recollection less frequently. In controls, eye movements at encoding predicted subsequent correct responses and subsequent recollection on a trial-by-trial basis, as expected. In contrast, despite a similar pattern of eye movements during encoding in the two groups, eye movements did not predict trial-by-trial subsequent memory in ASD. Furthermore, recollection was associated with lower similarity between encoding- and retrieval-related eye movements in the ASD group compared to the control group. The eye-tracking results therefore provide novel evidence for a dissociation between encoding and recollection-based retrieval in ASD.This research was supported by a James S. McDonnell Scholar Award to J.S.S., and an Economic and Social Research Council Award to R.A.C. We would like to thank all the participants who gave their time for this experiment, as well as the Autism Research Centre, Cambridge for helping with participant recruitment and providing access to the eye-tracking equipment used for this study

Scalar and vector Slepian functions, spherical signal estimation and spectral analysis

It is a well-known fact that mathematical functions that are timelimited (or spacelimited) cannot be simultaneously bandlimited (in frequency). Yet the finite precision of measurement and computation unavoidably bandlimits our observation and modeling scientific data, and we often only have access to, or are only interested in, a study area that is temporally or spatially bounded. In the geosciences we may be interested in spectrally modeling a time series defined only on a certain interval, or we may want to characterize a specific geographical area observed using an effectively bandlimited measurement device. It is clear that analyzing and representing scientific data of this kind will be facilitated if a basis of functions can be found that are "spatiospectrally" concentrated, i.e. "localized" in both domains at the same time. Here, we give a theoretical overview of one particular approach to this "concentration" problem, as originally proposed for time series by Slepian and coworkers, in the 1960s. We show how this framework leads to practical algorithms and statistically performant methods for the analysis of signals and their power spectra in one and two dimensions, and, particularly for applications in the geosciences, for scalar and vectorial signals defined on the surface of a unit sphere.Comment: Submitted to the 2nd Edition of the Handbook of Geomathematics, edited by Willi Freeden, Zuhair M. Nashed and Thomas Sonar, and to be published by Springer Verlag. This is a slightly modified but expanded version of the paper arxiv:0909.5368 that appeared in the 1st Edition of the Handbook, when it was called: Slepian functions and their use in signal estimation and spectral analysi

Reality monitoring impairment in schizophrenia reflects specific prefrontal cortex dysfunction

Reality monitoring impairment is often reported in schizophrenia but the neural basis of this deficit is poorly understood. Difficulties with reality monitoring could be attributable to the same pattern of neural dysfunction as other cognitive deficits that characterize schizophrenia, or might instead represent a separable and dissociable impairment. This question was addressed through direct comparison of behavioral performance and neural activity associated with reality monitoring and working memory in patients with schizophrenia and matched healthy controls. Participants performed a word-pair reality monitoring task and a Sternberg working memory task while undergoing fMRI scanning. Distinct behavioral deficits were observed in the patients during performance of each task, which were associated with separable task- and region-specific dysfunction in the medial anterior prefrontal cortex for reality monitoring and dorsolateral prefrontal cortex for working memory. The results suggest that reality monitoring impairment is a distinct neurocognitive deficit in schizophrenia. The findings are consistent with the presence of a range of dissociable cognitive deficits in schizophrenia which may be associated with variable functional and structural dysconnectivity in underlying processing networks.JRG was supported by a University of Cambridge Behavioural and Clinical Neuroscience Institute studentship, funded by a joint award from the UK Medical Research Council and the Wellcome Trust. JSS was supported by a James S. McDonnell Foundation Scholar award