Direct monitoring of breast and endometrial cancer cell epigenetic response to DNA methyltransferase and histone deacetylase inhibitors.

DNA methylation and histone deacetylation are key epigenetic processes involved in normal cellular function and tumorigenesis. Therapeutic strategies based on DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibitors are currently in use and under development for the treatment of cancers. Genome-wide DNA methylation profiling has been proposed for use in disease diagnosis, and histone modification profiling for disease stratification will follow suit. However, whether epigenome sequencing technologies will be feasible for rapid clinic diagnosis and patient treatment monitoring remains to be seen, and alternative detection technologies will almost certainly be needed. Here we used electrochemical impedance spectroscopy (EIS) employing a graphene-based screen-printed electrode system to directly measure global DNA methylation and histone H3 acetylation to compare non-cancer and breast cancer cell lines. We demonstrated that whilst global methylation was not useful as a differential marker in the cellular systems tested, histone H3 acetylation was effective at higher chromatin levels. Using breast and endometrial cancer cell models, EIS was then used to monitor cellular responses to the DNMT and HDAC inhibitors 5-Aza-2'-deoxycytidine and suberoylanilide hydroxamic acid in vitro, and proved very effective at detecting global cellular responses to either treatment, indicating that this approach could be useful in following treatment response to epigenetic drugs. Moreover, this work reports the first combined analysis of two epigenetic markers using a unified graphene-based biosensor platform, demonstrating the potential for multiplex analysis of both methylation and acetylation on the same sample.

Transcriptional signatures of connectomic subregions of the human striatum.

Functionally distinct regions of the brain are thought to possess a characteristic connectional fingerprint - a profile of incoming and outgoing connections that defines the function of that area. This observation has motivated efforts to subdivide brain areas using their connectivity patterns. However, it remains unclear whether these connectomically-defined subregions can be distinguished at the molecular level. Here, we combine high-resolution diffusion-weighted magnetic resonance imaging with transcriptomic data to show that connectomically-defined subregions of the striatum carry distinct transcriptional signatures. Using data-driven clustering of diffusion tractography, seeded from the striatum in 100 healthy individuals, we identify a tripartite organization of the caudate and putamen that comprises ventral, dorsal and caudal subregions. We then use microarray data of gene expression levels in 19 343 genes, taken from 98 tissue samples distributed throughout the striatum, to accurately discriminate the three connectomically-defined subregions with 80-90% classification accuracy using linear support vector machines. This classification accuracy was robust at the group and individual level and was superior for our parcellation of the striatum when compared with parcellations based on anatomical boundaries or other criteria. Genes contributing strongly to classification were enriched for gene ontology categories including dopamine signaling, glutamate secretion, response to amphetamine and metabolic pathways, and were implicated in risk for disorders such as schizophrenia, autism and Parkinson's disease. Our findings highlight a close link between regional variations in transcriptional activity and inter-regional connectivity in the brain, and suggest that there may be a strong genomic signature of connectomically-defined subregions of the brain.

Prevalence and subtypes of radiological cerebrovascular disease in late-onset isolated seizures and epilepsy.

BACKGROUND: Late-onset epilepsy (LOE), onset over 60, is often attributed to cerebrovascular disease (CVD), and is associated with increased stroke risk. We investigated the radiological prevalence of CVD in LOE. METHODS: We undertook a retrospective case-control study of patients with LOE and age and sex-matched controls, also matched for imaging modality. Radiological CVD was recorded, with radiological findings by an experienced consultant neuroradiologist usi a structured proforma. RESULTS: 105 cases and 105 controls were studied, comprising 61 (58.1%) males, mean (±SD) age (years) 72.7±7.48 (cases), 72.4±7.02 (controls). 9 cases had isolated seizures rather than LOE. Imaging modality (in cases and controls) was CT in 59 and MRI in 46. Radiological CVD was more prevalent amongst cases (65.7%) than controls (33.3%) (p<0.0001, Chi-square), odds ratio 3.83 (95% CI 2.16-6.79). Large vessel disease (LVD) (single or multiple cortical or subcortical infarcts>1.5 cm) was present in 23 (21.9%) cases and 2 (1.9%) (p<0.001) controls, with small vessel disease (SVD) (periventricular or subcortical white matter lesions (WMLs), including leukoaraiosis (LA)) present in 52 (49.5%) cases (LA in 4) and 34 (32.3%) controls (LA in 0) (p<0.05). When WMLs were rated using a semiquantitative visual rating scale, a trend towards greater severity was observed amongst cases compared to controls. CONCLUSIONS: Radiological CVD is significantly more prevalent in patients with LOE than controls, including signs of both LVD and SVD. SVD also appears to be of greater severity. Further studies are needed in this area.

The effect of sex and handedness on white matter anisotropy: a diffusion tensor magnetic resonance imaging study.

Diffusion tensor magnetic resonance imaging provides a way of assessing the asymmetry of white matter (WM) connectivity, the degree of anisotropic diffusion within a given voxel being a marker of coherently bundled myelinated fibers. Voxel-based statistical analysis was performed on fractional anisotropy (FA) images of 42 right- and 40 left-handers, to assess differences in underlying WM anisotropy and FA asymmetry across the whole brain. Right-handers show greater anisotropy than left-handers in the uncinate fasciculus (UF) within the limbic lobe, and WM underlying prefrontal cortex, medial and inferior frontal gyri. Significantly greater leftward FA asymmetry in cerebellum posterior lobe is seen in left- than right-handers, and males show significantly greater rightward (right-greater-than-left) FA asymmetry in regions of middle occipital lobe, medial temporal gyrus, and a region of the superior longitudinal fasciculus underlying the supramarginal gyrus. Leftward (left-greater-than-right) anisotropy is found in regions of the arcuate fasciculus (AF), UF, and WM underlying pars triangularis in both handedness groups, with right-handers alone showing additional leftward FA asymmetry along the length of the superior temporal gyrus. Overall results indicate that although both handedness groups show anisotropy in similar WM regions, greater anisotropy is observed in right-handers compared with left-handers. The largest differences in FA asymmetry are found between males and females, suggesting a greater effect of sex than handedness on FA asymmetry.

Retrospective, observational data collection of the treatment of phenylketonuria in the UK, and associated clinical and health outcomes.

OBJECTIVE: To determine the percentage of phenylketonuria (PKU) subjects using current treatment strategies whose phenylalanine (Phe) concentrations diverge from the UK target guidelines for PKU. RESEARCH DESIGN AND METHODS: This retrospective, observational, chart review was conducted between 2004 and 2008 at three specialist PKU treatment centres in the UK, and included 125 eligible subjects: 20 adults (18+ years, with ≥4 Phe concentrations measured per year) and 105 children (up to age 17, with ≥6 Phe concentrations measured per year). RESULTS: The mean percentage of subjects with at least 70% of Phe concentrations within the target range for 0-5-year olds, 6-10 year olds and 11-17 year olds was similar across the period 2004-2008 (57.0%, 56.5% and 57.1%, respectively) and lower (39.4%) in the 18+ year age group. For all ages, across the period the mean was 54.4%. Further analysis of the adult population showed that some subjects were very good at complying with treatment and reporting Phe concentrations. Overall, the percentage of 100% compliance was 15.7% in females and 13.7% in males. The mean duration that subjects were 'out of range' of target Phe concentrations over the study period was approximately 1 year and 3 months and the mean duration for 'significantly out of range' values was approximately 9 months. The most common type of contact made with subjects was by telephone, with a mean number of 16 calls per subject per year. CONCLUSION: The results support current literature showing that a proportion of subjects with PKU, in particular older subjects, are not fully compliant with their treatment and subsequently have Phe concentrations that depart from national recommendations. However, definitive conclusions may not be drawn due to the retrospective nature of the study and the small number of observed subjects.

Dynamic contrast-enhanced imaging techniques: CT and MRI.

Over the last few decades there has been considerable research into quantifying the cerebral microvasculature with imaging, for use in studies of the human brain and various pathologies including cerebral tumours. This review highlights key issues in dynamic contrast-enhanced CT, dynamic contrast-enhanced MRI and arterial spin labelling, the various applications of which are considered elsewhere in this special issue of the British Journal of Radiology.

Abnormalities of cerebral perfusion in multiple sclerosis.

BACKGROUND: Measuring perfusion provides a potential indication of metabolic activity in brain tissue. Studies in multiple sclerosis (MS) have identified areas of decreased perfusion in grey matter (GM) and white matter (WM), but the pattern in clinical subgroups is unclear. OBJECTIVES: This study investigated perfusion changes in differing MS clinical subgroups on or off beta-interferon therapy using a non-invasive MRI technique (continuous arterial spin labelling) to investigate whether different clinical MS subtypes displayed perfusion changes and whether this could give a further insight into the pathological mechanisms involved. METHODS: Sixty patients (21 relapsing remitting, 14 secondary progressive, 12 primary progressive, 13 benign) and 34 healthy controls were compared. Statistical parametric mapping (SPM '99) was used to investigate regional variations in perfusion in both GM and WM. Global WM perfusion was derived by segmenting WM from images using T(1) relaxation times. RESULTS: Regions of lower perfusion in predominantly GM were observed in the primary and secondary progressive cohorts, particularly in the thalamus. Increased WM perfusion was seen in relapsing remitting and secondary progressive cohorts. CONCLUSIONS: Low GM perfusion could reflect decreased metabolism secondary to neuronal and axonal loss or dysfunction with a predilection for progressive forms of MS. Increased WM perfusion may indicate increased metabolic activity possibly due to increased cellularity and inflammation. Improved methodology and longitudinal studies may enable further investigation of regional and temporal changes, and their relationship with physical and cognitive impairment.

Compulsory averaging of crowded orientation signals in human vision.

A shape can be more difficult to identify when other shapes are near it. For example, when several grating patches are viewed parafoveally, observers are unable to report the orientation of the central patch. This phenomenon, known as 'crowding,' has historically been confused with lateral masking, in which one stimulus attenuates signals generated by another stimulus. Here we show that despite their inability to report the orientation of an individual patch, observers can reliably estimate the average orientation, demonstrating that the local orientation signals are combined rather than lost. Our results imply that crowding is distinct from ordinary masking, and is perhaps related to texture perception. Under crowded conditions, the orientation signals in primary visual cortex are pooled before they reach consciousness.

Protection against group B meningococcal disease. I. Comparison of group-specific and type-specific protection in the chick embryo model.

Protection against group B meningococcal infection was examined using the chick embryo. 12-day-old embryos were challenged intravenously with various meningococcal strains. The chick embryo has an active reticuloendothelial system but lacks functional complement. In this model we found that protection against group B infection was primarily group specific. The group B polysaccharide antibody is an effective opsonin, but is a very poor bactericidal antibody. In contrast, the serotype antibody was bactericidal but only slightly protective in the chick embryo where protection is primarily phagocytic in nature. The group-specific and type-specific antibodies are strongly synergistic. Minute amounts of group B polysaccharide antibody caused a very significant increase in the protective effects of the serotype antibody.