Novel Assays of Thrombogenic Pathogenicity in the Antiphospholipid Syndrome Based on the Detection of Molecular Oxidative Modification of the Major Autoantigen β2-Glycoprotein I

Objective. Beta-2-glycoprotein I (beta(2)GPI) constitutes the major autoantigen in the antiphospholipid syndrome (APS), a common acquired cause of arterial and venous thrombosis. We recently described the novel observation that beta(2)GPI may exist in healthy individuals in a free thiol (biochemically reduced) form. The present study was undertaken to quantify the levels of total, reduced, and posttranslationally modified oxidized beta(2)GPI in APS patients compared to various control groups.Methods. In a retrospective multicenter analysis, the proportion of beta(2)GPI with free thiols in serum from healthy volunteers was quantified. Assays for measurement of reduced as well as total circulating beta(2)GPI were developed and tested in the following groups: APS (with thrombosis) (n = 139), autoimmune disease with or without persistent antiphospholipid antibodies (aPL) but without APS (n = 188), vascular thrombosis without APS or aPL (n = 38), and healthy volunteers (n = 91).Results. Total beta(2)GPI was significantly elevated in patients with APS (median 216.2 mu g/ml [interquartile range 173.3-263.8]) as compared to healthy subjects (median 178.4 mu g/ml [interquartile range 149.4-227.5] [P < 0.0002]) or control patients with autoimmune disease or vascular thrombosis (both P < 0.0001). The proportion of total beta(2)GPI in an oxidized form (i.e., lacking free thiols) was significantly greater in the APS group than in each of the 3 control groups (all P < 0.0001).Conclusion. This large retrospective multicenter study shows that posttranslational modification of beta(2)GPI via thiol-exchange reactions is a highly specific phenomenon in the setting of APS thrombosis. Quantification of posttranslational modifications of beta(2)GPI in conjunction with standard laboratory tests for APS may offer the potential to more accurately predict the risk of occurrence of a thrombotic event in the setting of APS

Seven HCI Grand Challenges

This article aims to investigate the Grand Challenges which arise in the current and emerging landscape of rapid technological evolution towards more intelligent interactive technologies, coupled with increased and widened societal needs, as well as individual and collective expectations that HCI, as a discipline, is called upon to address. A perspective oriented to humane and social values is adopted, formulating the challenges in terms of the impact of emerging intelligent interactive technologies on human life both at the individual and societal levels. Seven Grand Challenges are identified and presented in this article: Human-Technology Symbiosis; Human-Environment Interactions; Ethics, Privacy and Security; Well-being, Health and Eudaimonia; Accessibility and Universal Access; Learning and Creativity; and Social Organization and Democracy. Although not exhaustive, they summarize the views and research priorities of an international interdisciplinary group of experts, reflecting different scientific perspectives, methodological approaches and application domains. Each identified Grand Challenge is analyzed in terms of: concept and problem definition; main research issues involved and state of the art; and associated emerging requirements

Introduction of <it>in vitro </it>transcribed <it>ENO1 </it>mRNA into neuroblastoma cells induces cell death

Abstract Background Neuroblastoma is a solid tumour of childhood often with an unfavourable outcome. One common genetic feature in aggressive tumours is 1p-deletion. The α-enolase (ENO1) gene is located in chromosome region 1p36.2, within the common region of deletion in neuroblastoma. One alternative translated product of the ENO1 gene, known as MBP-1, acts as a negative regulator of the c-myc oncogene, making the ENO1 gene a candidate as a tumour suppressor gene. Methods Methods used in this study are transfection of cDNA-vectors and in vitro transcribed mRNA, cell growth assay, TUNEL-assay, real-time RT-PCR (TaqMan) for expression studies, genomic sequencing and DHPLC for mutation detection. Results Here we demonstrate that transfection of ENO1 cDNA into 1p-deleted neuroblastoma cell lines causes' reduced number of viable cells over time compared to a negative control and that it induces apoptosis. Interestingly, a similar but much stronger dose-dependent reduction of cell growth was observed by transfection of in vitro transcribed ENO1 mRNA into neuroblastoma cells. These effects could also be shown in non-neuroblastoma cells (293-cells), indicating ENO1 to have general tumour suppressor activity. Expression of ENO1 is detectable in primary neuroblastomas of all different stages and no difference in the level of expression can be detected between 1p-deleted and 1p-intact tumour samples. Although small numbers (11 primary neuroblastomas), there is some evidence that Stage 4 tumours has a lower level of ENO1-mRNA than Stage 2 tumours (p = 0.01). However, mutation screening of 44 primary neuroblastomas of all different stages, failed to detect any mutations. Conclusion Our studies indicate that ENO1 has tumour suppressor activity and that high level of ENO1 expression has growth inhibitory effects.</p

Apparent homozygosity of p.Phe508del in CFTR due to a large gene deletion of exons 4–11

We report a classic cystic fibrosis (CF) boy with a large deletion of exons 4–11 in the cystic fibrosis transmembrane conductance regulator (CFTR) gene on one allele and p.Phe508del in exon 10 on the second allele. Both parents of Georgian and Ukrainian background had no personal or family history of the disease. The initial molecular diagnostic investigation identified the patient as homozygous for the p.Phe508del and not compatible with his parent’s genetic status. The possibility of nonpaternity or uniparental disomy (UPD7) was investigated and excluded using microsatellite analysis of highly polymorphic markers on chromosome 7. Array-CGH was also performed on the patient and revealed a male profile with a subtle deletion within the CFTR gene on the long arm (q-arm) of chromosome 7 (7q31.2). The deletion was confirmed by MLPA extending from probe L02380 to probe L14978 (28.7 kb) and that was inherited from his father, while p.PheF508del was inherited from his mother. These data highlight the need for additional testing for large deletions in patients with apparent homozygosity for a mutated CFTR allele that do not match the carrier status of the parents. Not testing can lead to misdiagnosis and misinterpretation of mutation carrier status and the expected penetrance of the disorder

The Ledra Palace project: Using emerging technologies to communicate exhibition content - Evaluation of results

During the past couple of decades, museums resort to using innovative technological solutions in their permanent collections or temporary exhibitions aiming to enhance visitor experience. In the current study, we tested three emerging technologies (i.e., Interactive Book, Interactive Table, Immersive Virtual Reality) that were created to show in a museum exhibition, content related to ‘difficult heritage’ and ‘difficult history’. In a questionnaire administrated at the end of the exhibition, visitors were first asked to evaluate whether the content of the exhibition was better communicated through these interactive technologies than through non-technological (conventional) installations, and then to assess the usability of these technologies. Results revealed that technological installations were as engaging and successful in the presentation and communication of the content of the exhibition as the non-technological installations. Finally, with respect to the usability of the above three technological installations, results were remarkably high (Mdn: 87), with all visitors reporting a clear preference for the immersive virtual reality installation. Studies such as the current one support that interactive technologies should not aim to substitute conventional installations but instead, to complement them to enhance visitor experience and provide alternativ

A new bacteriophage P1-derived vector for the propagation of large human DNA fragments

We have designed a P1 vector (pCYPAC−1) for the introduction of recombinant DNA into E. coli using electroporation procedures. The new cloning system, P1−derived arteficial chromosomes (PACs), was used to establish an initial 15,000 clone library with an average insert size of 130−150 kilobase pairs (kb). No chimaerism has been observed in 34 clones, by fluorescence in situ hybridization. Similarly, no insert instability has been observed after extended culturing, for 20 clones. We conclude that the PAC cloning system will be useful in the mapping and detailed analysis of complex genomes

-mut and -constructs with translation starts (ATG) and mutated (ATG→ATC) sites in -mut at positions 374 and 383 indicated

<p><b>Copyright information:</b></p><p>Taken from "Introduction of transcribed mRNA into neuroblastoma cells induces cell death"</p><p>BMC Cancer 2005;5():161-161.</p><p>Published online 16 Dec 2005</p><p>PMCID:PMC1327688.</p><p>Copyright © 2005 Ejeskär et al; licensee BioMed Central Ltd.</p> The mutations introduce I and I restriction enzyme sites respectively. Size determination of cDNA-constructs in pIRES-EGFP-vector by I/I-digest. Confirmation of mutations in -mut DNA by I and I digests