Untemplated Oligoadenylation Promotes Degradation of RISC-Cleaved Transcripts

In the best-characterized mechanism of RNAmediated silencing, small interfering RNAs (siRNAs), incorporated into the RNA-induced silencing complex (RISC), guide the endonucleolytic cleavage of complementary RNAs (1). In Drosophila melanogaster, these RISC-generated products are eventually degraded by exoribonucleases: Xrn1, a 5′-to-3′ exonuclease, and exosome, a 3′-to-5′ multisubunit exonuclease (2). Interestingly, in Arabidopsis thaliana and in mammals, an oligouridine or oligoadenine [oligo(U/A)] tail is added to the 5′ RNA fragments resulting from microRNA-directed cleavage (3). However, the biological role of this tail remains unclear

Pathways to school readiness: Executive functioning predicts academic and social-emotional aspects of school readiness.

The current study specified the extent to which hot and cool aspects of executive functioning predicted academic and social-emotional indicators of school readiness. It was unique in focusing on positive aspects of social-emotional readiness, rather than problem behaviors. One hundred four 3- to 5-year-old children completed tasks measuring executive functioning, social-emotional readiness, academic readiness, and vocabulary. As expected, age predicted executive functioning components and social-emotional readiness. Moreover, working memory and inhibitory control directly predicted academic readiness, whereas delay of gratification predicted social-emotional readiness. Working memory and inhibitory control predicted delay of gratification, consistent with the notion that simpler executive functions may set the stage for more complex executive functions. Interestingly, social-emotional readiness predicted academic readiness. These findings confirm that hot and cool aspects of executive functioning are related to social-emotional and academic school readiness

Novel TNF Receptor-1 Inhibitors Identified as Potential Therapeutic Candidates for Traumatic Brain Injury

Background: Traumatic brain injury (TBI) begins with the application of mechanical force to the head or brain, which initiates systemic and cellular processes that are hallmarks of the disease. The pathological cascade of secondary injury processes, including inflammation, can exacerbate brain injury-induced morbidities and thus represents a plausible target for pharmaceutical therapies. We have pioneered research on post-traumatic sleep, identifying that injury-induced sleep lasting for 6 h in brain-injured mice coincides with increased cortical levels of inflammatory cytokines, including tumor necrosis factor (TNF). Here, we apply post-traumatic sleep as a physiological bio-indicator of inflammation. We hypothesized the efficacy of novel TNF receptor (TNF-R) inhibitors could be screened using post-traumatic sleep and that these novel compounds would improve functional recovery following diffuse TBI in the mouse. Methods: Three inhibitors of TNF-R activation were synthesized based on the structure of previously reported TNF CIAM inhibitor F002, which lodges into a defined TNFR1 cavity at the TNF-binding interface, and screened for in vitro efficacy of TNF pathway inhibition (IκB phosphorylation). Compounds were screened for in vivo efficacy in modulating post-traumatic sleep. Compounds were then tested for efficacy in improving functional recovery and verification of cellular mechanism. Results: Brain-injured mice treated with Compound 7 (C7) or SGT11 slept significantly less than those treated with vehicle, suggesting a therapeutic potential to target neuroinflammation. SGT11 restored cognitive, sensorimotor, and neurological function. C7 and SGT11 significantly decreased cortical inflammatory cytokines 3 h post-TBI. Conclusions: Using sleep as a bio-indicator of TNF-R-dependent neuroinflammation, we identified C7 and SGT11 as potential therapeutic candidates for TBI

Identifying the motivations of African American volunteers working to prevent HIV/AIDS

Community-based organizations that are engaged in HIV/AIDS prevention and support services often rely on volunteers. This article describes the development of a 22-item inventory that measures the motivations of volunteers who deliver HIV prevention education in the African American community. In a statewide survey of volunteers (N = 102), the two strongest motivations for volunteer activity were concern for the African American community and a desire to understand the causes and consequences of the epidemic. These motives predicted the frequency that volunteers held discussions about HIV/AIDS with members of their community. Discussion focuses on the relevance of the results for the recruitment, training, and retention of volunteers

The novel RASSF6 and RASSF10 candidate tumour suppressor genes are frequently epigenetically inactivated in childhood leukaemias.

BACKGROUND: The Ras-association family (RASSF) of tumour suppressor genes (TSGs) contains 10 members that encode proteins containing Ras-association (RA) domains. Several members of the RASSF family are frequently epigenetically inactivated in cancer, however, their role in leukaemia has remained largely uninvestigated. Also, RASSF10 is a predicted gene yet to be experimentally verified. Here we cloned, characterised and demonstrated expression of RASSF10 in normal human bone marrow. We also determined the methylation status of CpG islands associated with RASSF1-10 in a series of childhood acute lymphocytic leukaemias (ALL) and normal blood and bone marrow samples. RESULTS: COBRA and bisulphite sequencing revealed RASSF6 and RASSF10 were the only RASSF members with a high frequency of leukaemia-specific methylation. RASSF6 was methylated in 94% (48/51) B-ALL and 41% (12/29) T-ALL, whilst RASSF10 was methylated in 16% (8/51) B-ALL and 88% (23/26) T-ALL. RASSF6 and RASSF10 expression inversely correlated with methylation which was restored by treatment with 5-aza-2'deoxycytidine (5azaDC). CONCLUSION: This study shows the hypermethylation profile of RASSF genes in leukaemias is distinct from that of solid tumours and represents the first report of inactivation of RASSF6 or RASSF10 in cancer. These data show epigenetic inactivation of the candidate TSGs RASSF6 and RASSF10 is an extremely frequent event in the pathogenesis of childhood leukaemia. This study also warrants further investigation of the newly identified RASSF member RASSF10 and its potential role in leukaemia.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

Scalable Transcriptome Preparation for Massive Parallel Sequencing

Background: The tremendous output of massive parallel sequencing technologies requires automated robust and scalable sample preparation methods to fully exploit the new sequence capacity. Methodology: In this study, a method for automated library preparation of RNA prior to massively parallel sequencing is presented. The automated protocol uses precipitation onto carboxylic acid paramagnetic beads for purification and size selection of both RNA and DNA. The automated sample preparation was compared to the standard manual sample preparation. Conclusion/Significance: The automated procedure was used to generate libraries for gene expression profiling on the Illumina HiSeq 2000 platform with the capacity of 12 samples per preparation with a significantly improved throughput compared to the standard manual preparation. The data analysis shows consistent gene expression profiles in terms of sensitivity and quantification of gene expression between the two library preparation methods

RASSF2, a potential tumour suppressor, is silenced by CpG island hypermethylation in gastric cancer

RASSF2, a member of the RASSF1 family, has recently been identified as a potential tumour suppressor. We examined methylation status in multiple regions which included the CpG island and spanned the transcription start site of RASSF2 in 10 gastric cancer cell lines, as well as 78 primary gastric cancers and corresponding non-neoplastic gastric epithelia. Hypermethylation of RASSF2 in at least one of the regions examined was detected in seven (70%) of the 10 cell lines; two (20%) exhibited hypermethylation in all the regions examined including the transcription start site and lost expression of RASSF2 mRNA, which could, however, be restored by 5-aza-2′ deoxycytidine treatment, while the other five (50%) cell lines exhibited hypermethylation at the 5′- and/or 3′- edge, with four of them expressing RASSF2 mRNA. In primary gastric cancers and corresponding non-neoplastic gastric epithelia, frequencies of RASSF2 methylation ranged from 29% (23 out of 78) to 79% (62 out of 78) and 3% (two out of 78) to 60% (47 out of 78), respectively, at different CpG sites examined. Methylation was frequently observed at the 5′- and 3′- edges, and became less frequent near the transcription start site in both the primary gastric cancers and corresponding non-neoplastic gastric epithelia. Hypermethylation near the transcription start site was mostly cancer-specific. We thus showed that RASSF2 is silenced by hypermethylation near the transcription start site in gastric cancer. Hypermethylation was found initially to occur at the 5′- and 3′- furthest regions of the CpG island in non-neoplastic gastric epithelia, to gradually spreads near the transcription start site to shut down RASSF2 expression, and ultimately to constitute a field-defect placing tissue increased risk for development of gastric cancer

An electrochemical system for efficiently harvesting low-grade heat energy

Efficient and low-cost thermal energy-harvesting systems are needed to utilize the tremendous low-grade heat sources. Although thermoelectric devices are attractive, its efficiency is limited by the relatively low figure-of-merit and low-temperature differential. An alternative approach is to explore thermodynamic cycles. Thermogalvanic effect, the dependence of electrode potential on temperature, can construct such cycles. In one cycle, an electrochemical cell is charged at a temperature and then discharged at a different temperature with higher cell voltage, thereby converting heat to electricity. Here we report an electrochemical system using a copper hexacyanoferrate cathode and a Cu/Cu2+ anode to convert heat into electricity. The electrode materials have low polarization, high charge capacity, moderate temperature coefficients and low specific heat. These features lead to a high heat-to-electricity energy conversion efficiency of 5.7% when cycled between 10 and 60 degrees C, opening a promising way to utilize low-grade heat.open121

Discourse markers activate their, <i>like</i>, cohort competitors

Speech in everyday conversations is riddled with discourse markers (DMs), such as well, you know, and like. However, in many lab-based studies of speech comprehension, such DMs are typically absent from the carefully articulated and highly controlled speech stimuli. As such, little is known about how these DMs influence online word recognition. The present study specifically investigated the online processing of DM like and how it influences the activation of words in the mental lexicon. We specifically targeted the cohort competitor (CC) effect in the Visual World Paradigm: Upon hearing spoken instructions to “pick up the beaker,” human listeners also typically fixate—next to the target object—referents that overlap phonologically with the target word (cohort competitors such as beetle; CCs). However, several studies have argued that CC effects are constrained by syntactic, semantic, pragmatic, and discourse constraints. Therefore, the present study investigated whether DM like influences online word recognition by activating its cohort competitors (e.g., lightbulb). In an eye-tracking experiment using the Visual World Paradigm, we demonstrate that when participants heard spoken instructions such as “Now press the button for the, like … unicycle,” they showed anticipatory looks to the CC referent (lightbulb)well before hearing the target. This CC effect was sustained for a relatively long period of time, even despite hearing disambiguating information (i.e., the /k/ in like). Analysis of the reaction times also showed that participants were significantly faster to select CC targets (lightbulb) when preceded by DM like. These findings suggest that seemingly trivial DMs, such as like, activate their CCs, impacting online word recognition. Thus, we advocate a more holistic perspective on spoken language comprehension in naturalistic communication, including the processing of DMs