Core binding factor (CBF) is required for Epstein-Barr virus EBNA3 proteins to regulate target gene expression

ChIP-seq performed on lymphoblastoid cell lines (LCLs), expressing epitope-tagged EBNA3A, EBNA3B or EBNA3C from EBV-recombinants, revealed important principles of EBNA3 binding to chromatin. When combined with global chromatin looping data, EBNA3-bound loci were found to have a singular character, each directly associating with either EBNA3-repressed or EBNA3-activated genes, but not with both. EBNA3A and EBNA3C showed significant association with repressed and activated genes. Significant direct association for EBNA3B loci could only be shown with EBNA3B-repressed genes. A comparison of EBNA3 binding sites with known transcription factor binding sites in LCL GM12878 revealed substantial co-localization of EBNA3s with RUNX3-a protein induced by EBV during B cell transformation. The beta-subunit of core binding factor (CBFβ), that heterodimerizes with RUNX3, could co-immunoprecipitate robustly EBNA3B and EBNA3C, but only weakly EBNA3A. Depletion of either RUNX3 or CBFβ with lentivirus-delivered shRNA impaired epitope-tagged EBNA3B and EBNA3C binding at multiple regulated gene loci, indicating a requirement for CBF heterodimers in EBNA3 recruitment during target-gene regulation. ShRNA-mediated depletion of CBFβ in an EBNA3C-conditional LCL confirmed the role of CBF in the regulation of EBNA3C-induced and -repressed genes. These results reveal an important role for RUNX3/CBF during B cell transformation and EBV latency that was hitherto unexplored

The effect of increased microporosity on bone formation within silicate-substituted scaffolds in an ovine posterolateral spinal fusion model

This study compared the bone forming capacity of the same formulation of silicate-substituted bone graft substitute materials with different microporosity in an instrumented posterolateral spinal fusion ovine model. Materials with a strut porosity of (i) 22.5% (SiCaP) or (ii) 36.0% (SiCaP(+)) were packed along either side of the spine. Bone apposition rates, % new bone formation, % bone-implant contact, and % graft resorption were quantified at 8, 12, and 24 weeks post surgery. Computed Tomography (CT) was used to grade the formation of fusion bridges between vertebrae. Results showed no significant difference in bone apposition rates, % new bone formation, and % bone-implant contact when the two materials were compared. However, at 8 weeks, a significantly higher CT score was obtained in the SiCaP(+) group (0.83±0.17) when compared with the SiCaP group (0.17±0.17; p=0.027). Significantly less scaffold remained in the SiCaP(+) group at 12 weeks (p=0.018). Both SiCaP and SiCaP(+) formulations augmented bone formation. Increasing the strut porosity did not significantly increase bone formation however, at 8 weeks it promoted the formation of more highly mineralized bone resulting in a significantly higher CT score, suggesting the bone tissue formed was more mature

High Throughput Screening of a GlaxoSmithKline Protein Kinase Inhibitor Set Identifies an Inhibitor of Human Cytomegalovirus Replication that Prevents CREB and Histone H3 Post-Translational Modification.

To identify new compounds with anti-human cytomegalovirus (HCMV) activity and new anti-HCMV targets, we developed a high throughput strategy to screen a GlaxoSmithKline (GSK) Published Kinase Inhibitor Set (PKIS). This collection contains a range of extensively characterized compounds grouped into chemical families (chemotypes). From our screen we identified compounds within chemotypes that impede HCMV replication and identified kinase proteins associated with inhibition of HCMV replication that are potential novel anti-HCMV targets. We focused our study on a top "hit" in our screen, SB-734117, which we found inhibits productive replication of several HCMV strains. Kinase selectivity data indicated that SB-734117 exhibits polypharmacology and is an inhibitor of several proteins from the AGC and CMCG kinase groups. Using western blotting we found that SB-734711 inhibited accumulation of HCMV immediate-early proteins, phosphorylation of cellular proteins involved in immediate-early protein production (CREB and histone H3) and histone H3 lysine 36 trimethylation (H3K36me3). Therefore, we identify SB-734117 as a novel anti-HCMV compound and find that inhibition of AGC and CMCG kinase proteins during productive HCMV replication is associated with inhibition of viral protein production and prevents post-translational modification of cellular factors associated with viral protein production

Use of indigenous knowledge in the management of field and storage pests around Lake Victoria basin in Tanzania

Agriculture in Lake Victoria basin (LVB) in Tanzania is predominantly subsistence and is characterised by perennial food deficits, cyclic famines and poverty prompted largely by unreliable rainfall patterns, declining soil fertility and food grains pests and diseases. The pest problem is more pronounced as farmers are yet to fully integrate synthetic pesticides into their insect pest management systems due to subsistence nature of production and high poverty levels that make them rely on indigenous knowledge (IK) systems to meet their needs. The survey was conducted to document farmers’ IK on management of key field and storage insect pests in Magu and Misungwi districts in the LVB, Tanzania. Major crops grown were maize, rice, sorghum, finger millet, bean, groundnut, cowpea, green gram, brassicas, chicken pea, cassava, sweet potato, cotton and vegetables. Crops were mainly infested by Busseola fusca (Lepidoptera: Noctuidae), Spodoptera spp (Lepidoptera: Noctuidae), Agrotis spp (Lepidoptera: Noctuidae), Maruca vitrata (Lepidoptera: Crambidae), Rhopalosiphum maidis (Homoptera: Aphididae), Aphis fabae (Hemiptera: Aphididae), and grasshoppers in field and Stophilus spp (Coleoptera: Curculionidae), Prostephanus truncates (Coleoptera: Bostrichidae), Tribolium spp (Coleoptera: Tenebrionidae), Bruchus rufimanus (Coleoptera; Bruchidae), Rhyzopertha dominica (Coleoptera: Bostrichidae) and rodents on storage. IK based control methods used by farmers ranged from animal by-products (cow’s urine and dung), plant parts (Azadirachta indica (Meliaceae),Tephrosia vogelii (Fabaceae), Tamarindus indica (Fabaceae), Aloe spp (Asphodelaceae), red pepper, Capsicum spp (Solanaceae), Nicotiana tabasum (Solanaceae) to ash (general and specific) in the field. They also used neem, Chenopodium opulifolium (Chenopodiaceae), Ocimum suave (Labiatae), Senna siamea (Fabaceae or Caesalpinioideae), tobacco and Eucalyptus spp (Myrtaceae) and plant by-products (rice husks, ash from rice husks and red maize cobs and general ash) to control storage pests. Most of these products were used together with one or two others in different formulation mixtures. However, the formulations had variable amount taken during preparation, crop/ crop product treated, preparation times, modes and rates of application. Research is needed to unveil the amount for mixing, appropriate treatment, and application rate to ensure optimum concentration for specific pest. To ensure quality and safety, biosafety and quality studies are required for quality assessment of resulting product for human health. For understanding of active compounds in the formulations, chemical composition analysis of properly prepared solutions is required. Key words: Field and storage pests, indigenous knowledge, Tanzania, botanical formulation, Lake Victoria basin

Significance of herpesvirus immediate early gene expression in cellular immunity to cytomegalovirus infection

Interstitial pneumonia linked with reactivation of latent human cytomegalovirus due to iatrogenic immunosuppression can be a serious complication of bone marrow transplantation therapy of aplastic anaemia and acute leukaemia1. Cellular immunity plays a critical role in the immune surveillance of inapparent cytomegalovirus infections in man and the mouse1−7. The molecular basis of latency, however, and the interaction between latently or recurrently infected cells and the immune system of the host are poorfy understood. We have detected a so far unknown antigen in the mouse model. This antigen is found in infected cells in association with the expression of the herpesvirus 'immediate early' genes and is recognized by cytolytic T lymphocytes (CTL)8. We now demonstrate that an unexpectedly high proportion of the CTL precursors generated in vivo during acute murine cytomegalovirus infection are specific for cells that selectively synthesize immediate early proteins, indicating an immunodominant role of viral non-structural proteins

The prognosis of allocentric and egocentric neglect : evidence from clinical scans

We contrasted the neuroanatomical substrates of sub-acute and chronic visuospatial deficits associated with different aspects of unilateral neglect using computed tomography scans acquired as part of routine clinical diagnosis. Voxel-wise statistical analyses were conducted on a group of 160 stroke patients scanned at a sub-acute stage. Lesion-deficit relationships were assessed across the whole brain, separately for grey and white matter. We assessed lesions that were associated with behavioural performance (i) at a sub-acute stage (within 3 months of the stroke) and (ii) at a chronic stage (after 9 months post stroke). Allocentric and egocentric neglect symptoms at the sub-acute stage were associated with lesions to dissociated regions within the frontal lobe, amongst other regions. However the frontal lesions were not associated with neglect at the chronic stage. On the other hand, lesions in the angular gyrus were associated with persistent allocentric neglect. In contrast, lesions within the superior temporal gyrus extending into the supramarginal gyrus, as well as lesions within the basal ganglia and insula, were associated with persistent egocentric neglect. Damage within the temporo-parietal junction was associated with both types of neglect at the sub-acute stage and 9 months later. Furthermore, white matter disconnections resulting from damage along the superior longitudinal fasciculus were associated with both types of neglect and critically related to both sub-acute and chronic deficits. Finally, there was a significant difference in the lesion volume between patients who recovered from neglect and patients with chronic deficits. The findings presented provide evidence that (i) the lesion location and lesion size can be used to successfully predict the outcome of neglect based on clinical CT scans, (ii) lesion location alone can serve as a critical predictor for persistent neglect symptoms, (iii) wide spread lesions are associated with neglect symptoms at the sub-acute stage but only some of these are critical for predicting whether neglect will become a chronic disorder and (iv) the severity of behavioural symptoms can be a useful predictor of recovery in the absence of neuroimaging findings on clinical scans. We discuss the implications for understanding the symptoms of the neglect syndrome, the recovery of function and the use of clinical scans to predict outcome