The utility of computed tomography for recent-onset partial seizures in childhood

Objectives. The objective was to evaluate the role of a paediatric surgical consultant at a primary health care facility. Design. Descriptive and prospective. Setting. In the process of planning and implementation of the 2010 health plan of the Provincial Government of the Western Cape, a shift occurred in the delivery of health care to children from a provincially based hospital system to a municipally based primary health care system. To contribute towards enabling this process, the Department of Paediatric Surgery at Red Cross War Memorial Children’s Hospital established a paediatric surgical day clinic at a local community health centre during 2001. Subjects. Information was obtained from patient data sheets containing details of consultations at the sub-specialist surgical clinic at Michael Mapongwana Community Health Centre. Results. Over a 58-month period 1 171 children were seen, of whom 655 were male and 427 female. Their ages ranged from 0 to 19 years, the largest group being under 1 year. Eighty per cent of patients were accompanied by their mothers. The correct diagnosis was established by the nurse practitioners in 71%. General paediatric surgical conditions predominated, followed by medical, dermatological, orthopaedic, trauma, otolaryngo-pharyngology, infectious diseases, ophthalmology, urology, neurosurgery, malignancy and maxillofacial conditions. The details are set out in the report. In total 597 patients were referred directly to an appropriate care facility and 574 patients could be managed entirely at the clinic level. Conclusions. This study demonstrated the significant public health problem of paediatric surgical disease. It emphasised the preventative and cost-effective role of a surgical clinic at primary health care level. The clinic allowed for timely surgical intervention in 65% of surgical cases, thereby decreasing inappropriate tertiary referrals. We believe that bringing specialists into the community can only strengthen the 2010 health care plan

Longitudinal associations between conflict monitoring and emergent academic skills: An event‐related potentials study

Identifying the links between specific cognitive functions and emergent academic skills can help determine pathways to support both early academic performance and later academic achievement. Here, we investigated the longitudinal associations between a key aspect of cognitive control, conflict monitoring, and emergent academic skills from preschool through first grade, in a large sample of socioeconomically diverse children (N = 261). We recorded event‐related potentials (ERPs) during a Go/No‐Go task. The neural index of conflict monitoring, ΔN2, was defined as larger N2 mean amplitudes for No‐Go versus Go trials. ΔN2 was observed over the right hemisphere across time points and showed developmental stability. Cross‐lagged panel models revealed prospective links from ΔN2 to later math performance, but not reading performance. Specifically, larger ΔN2 at preschool predicted higher kindergarten math performance, and larger ΔN2 at kindergarten predicted higher first‐grade math performance, above and beyond the behavioral performance in the Go/No‐Go task. Early academic skills did not predict later ΔN2. These findings provided electrophysiological evidence for the contribution of conflict monitoring abilities to emergent math skills. In addition, our findings suggested that neural indices of cognitive control can provide additional information in predicting emergent math skills, above and beyond behavioral task performance.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/149228/1/dev21809.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/149228/2/dev21809_am.pd

Detecting new microRNAs in human osteoarthritic chondrocytes identifies miR-3085 as a human, chondrocyte-selective, microRNA

Objective: To use deep sequencing to identify novel microRNAs in human osteoarthritic cartilage which have a functional role in chondrocyte phenotype or function. Design: A small RNA library was prepared from human osteoarthritic primary chondrocytes using in-house adaptors and analysed by Illumina sequencing. Novel candidate microRNAs were validated by northern blot and qRT-PCR. Expression was measured in cartilage models. Targets of novel candidates were identified by microarray and computational analysis, validated using 3’-UTR-luciferase reporter plasmids. Protein levels were assessed by western blot and functional analysis by cell adhesion. Results: We identified 990 known microRNAs and 1621 potential novel microRNAs in human osteoarthritic chondrocytes, 60 of the latter were expressed in all samples assayed. MicroRNA-140-3p was the most highly expressed microRNA in osteoarthritic cartilage. Sixteen novel candidate microRNAs were analysed further, of which 6 remained after northern blot analysis. Three novel microRNAs were regulated across models of chondrogenesis, chondrocyte differentiation or cartilage injury. One sequence (novel #11), annotated in rodents as microRNA-3085-3p, was preferentially expressed in cartilage, dependent on chondrocyte differentiation and, in man, is located in an intron of the cartilage-expressed gene CRTAC-1. This microRNA was shown to target the ITGA5 gene directly (which encodes integrin alpha5) and inhibited adhesion to fibronectin (dependent on alpha5beta1 integrin). Conclusion: Deep sequencing has uncovered many potential microRNA candidates expressed in human cartilage. At least three of these show potential functional interest in cartilage homeostasis and osteoarthritis. Particularly, novel #11 (microRNA-3085-3p) which has been identified for the first time in man

The PRH/Hex repressor protein causes nuclear retention of Groucho/TLE co-repressors

The Proline-Rich Homeodomain protein (PRH/Hex) is a transcription factor that functions as an important regulator of vertebrate development and many other processes in the adult including haematopoiesis. The Groucho/TLE family of co-repressor proteins also regulate development and modulate the activity of many DNA-binding transcription factors during a range of diverse cellular processes including haematopoiesis. We have shown previously that PRH is a repressor of transcription in haematopoietic cells and that an Eh-1 motif present within the N-terminal transcription repression domain of PRH mediates binding to Groucho/TLE proteins and enables co-repression. Here we demonstrate that PRH regulates the nuclear retention of TLE proteins during cellular fractionation. We show that transcriptional repression and the nuclear retention of TLE proteins requires PRH to bind to both TLE and DNA. In addition, we characterise a trans-dominant negative PRH protein that inhibits wild type PRH activity by sequestering TLE proteins to specific sub-nuclear domains. These results demonstrate that transcriptional repression by PRH is dependent on TLE availability and suggest that sub-nuclear localisation of TLE plays an important role in transcriptional repression by PRH

Microguards and micromessengers of the genome

The regulation of gene expression is of fundamental importance to maintain organismal function and integrity and requires a multifaceted and highly ordered sequence of events. The cyclic nature of gene expression is known as ‘transcription dynamics’. Disruption or perturbation of these dynamics can result in significant fitness costs arising from genome instability, accelerated ageing and disease. We review recent research that supports the idea that an important new role for small RNAs, particularly microRNAs (miRNAs), is in protecting the genome against short-term transcriptional fluctuations, in a process we term ‘microguarding’. An additional emerging role for miRNAs is as ‘micromessengers’—through alteration of gene expression in target cells to which they are trafficked within microvesicles. We describe the scant but emerging evidence that miRNAs can be moved between different cells, individuals and even species, to exert biologically significant responses. With these two new roles, miRNAs have the potential to protect against deleterious gene expression variation from perturbation and to themselves perturb the expression of genes in target cells. These interactions between cells will frequently be subject to conflicts of interest when they occur between unrelated cells that lack a coincidence of fitness interests. Hence, there is the potential for miRNAs to represent both a means to resolve conflicts of interest, as well as instigate them. We conclude by exploring this conflict hypothesis, by describing some of the initial evidence consistent with it and proposing new ideas for future research into this exciting topic

The microRNA-29 family in cartilage homeostasis and osteoarthritis

MicroRNAs have been shown to function in cartilage development and homeostasis, as well as in progression of osteoarthritis. The objective of the current study was to identify microRNAs involved in the onset or early progression of osteoarthritis and characterise their function in chondrocytes. MicroRNA expression in mouse knee joints post-DMM surgery was measured over 7 days. Expression of miR-29b-3p was increased at day 1 and regulated in the opposite direction to its potential targets. In a mouse model of cartilage injury and in end-stage human OA cartilage, the miR-29 family were also regulated. SOX9 repressed expression of miR-29a-3p and miR-29b-3p via the 29a/b1 promoter. TGFβ1 decreased expression of miR-29a, b and c (3p) in primary chondrocytes, whilst IL-1β increased (but LPS decreased) their expression. The miR-29 family negatively regulated Smad, NFκB and canonical WNT signalling pathways. Expression profiles revealed regulation of new WNT-related genes. Amongst these, FZD3, FZD5, DVL3, FRAT2, CK2A2 were validated as direct targets of the miR-29 family. These data identify the miR-29 family as microRNAs acting across development and progression of OA. They are regulated by factors which are important in OA and impact on relevant signalling pathways

Infected Cell Killing by HIV-1 Protease Promotes NF-κB Dependent HIV-1 Replication

Acute HIV-1 infection of CD4 T cells often results in apoptotic death of infected cells, yet it is unclear what evolutionary advantage this offers to HIV-1. Given the independent observations that acute T cell HIV-1 infection results in (1) NF-κB activation, (2) caspase 8 dependent apoptosis, and that (3) caspase 8 directly activates NF-κB, we questioned whether these three events might be interrelated. We first show that HIV-1 infected T cell apoptosis, NF-κB activation, and caspase 8 cleavage by HIV-1 protease are coincident. Next we show that HIV-1 protease not only cleaves procaspase 8, producing Casp8p41, but also independently stimulates NF-κB activity. Finally, we demonstrate that the HIV protease cleavage of caspase 8 is necessary for optimal NF-κB activation and that the HIV-1 protease specific cleavage fragment Casp8p41 is sufficient to stimulate HIV-1 replication through NF-κB dependent HIV-LTR activation both in vitro as well as in cells from HIV infected donors. Consequently, the molecular events which promote death of HIV-1 infected T cells function dually to promote HIV-1 replication, thereby favoring the propagation and survival of HIV-1

Estimation of hydraulic conductivity and its uncertainty from grain-size data using GLUE and artificial neural networks

peer reviewedaudience: researcher, professionalVarious approaches exist to relate saturated hydraulic conductivity (Ks) to grain-size data. Most methods use a single grain-size parameter and hence omit the information encompassed by the entire grain-size distribution. This study compares two data-driven modelling methods, i.e.multiple linear regression and artificial neural networks, that use the entire grain-size distribution data as input for Ks prediction. Besides the predictive capacity of the methods, the uncertainty associated with the model predictions is also evaluated, since such information is important for stochastic groundwater flow and contaminant transport modelling. Artificial neural networks (ANNs) are combined with a generalized likelihood uncertainty estimation (GLUE) approach to predict Ks from grain-size data. The resulting GLUE-ANN hydraulic conductivity predictions and associated uncertainty estimates are compared with those obtained from the multiple linear regression models by a leave-one-out cross-validation. The GLUE-ANN ensemble prediction proved to be slightly better than multiple linear regression. The prediction uncertainty, however, was reduced by half an order of magnitude on average, and decreased at most by an order of magnitude. This demonstrates that the proposed method outperforms classical data-driven modelling techniques. Moreover, a comparison with methods from literature demonstrates the importance of site specific calibration. The dataset used for this purpose originates mainly from unconsolidated sandy sediments of the Neogene aquifer, northern Belgium. The proposed predictive models are developed for 173 grain-size -Ks pairs. Finally, an application with the optimized models is presented for a borehole lacking Ks data

HIV-1 Nef Induces Proinflammatory State in Macrophages through Its Acidic Cluster Domain: Involvement of TNF Alpha Receptor Associated Factor 2

Background: HIV-1 Nef is a virulence factor that plays multiple roles during HIV replication. Recently, it has been described that Nef intersects the CD40 signalling in macrophages, leading to modification in the pattern of secreted factors that appear able to recruit, activate and render T lymphocytes susceptible to HIV infection. The engagement of CD40 by CD40L induces the activation of different signalling cascades that require the recruitment of specific tumor necrosis factor receptor-associated factors (i.e. TRAFs). We hypothesized that TRAFs might be involved in the rapid activation of NF-kappa B, MAPKs and IRF-3 that were previously described in Nef-treated macrophages to induce the synthesis and secretion of proinflammatory cytokines, chemokines and IFN beta to activate STAT1, -2 and -3. Methodology/Principal Findings: Searching for possible TRAF binding sites on Nef, we found a TRAF2 consensus binding site in the AQEEEE sequence encompassing the conserved four-glutamate acidic cluster. Here we show that all the signalling effects we observed in Nef treated macrophages depend on the integrity of the acidic cluster. In addition, Nef was able to interact in vitro with TRAF2, but not TRAF6, and this interaction involved the acidic cluster. Finally silencing experiments in THP-1 monocytic cells indicate that both TRAF2 and, surprisingly, TRAF6 are required for the Nef-induced tyrosine phosphorylation of STAT1 and STAT2. Conclusions: Results reported here revealed TRAF2 as a new possible cellular interactor of Nef and highlighted that in monocytes/macrophages this viral protein is able to manipulate both the TRAF/NF-kappa B and TRAF/IRF-3 signalling axes, thereby inducing the synthesis of proinflammatory cytokines and chemokines as well as IFN beta