927 resultados para Phosphorylation sites


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Matrix Metalloproteinases (MMP) play a key role in osteoarthritis (OA) development. The aim of the present study was to investigate whether, the cross-talk between subchondral bone osteoblasts (SBOs) and articular cartilage chondrocytes (ACCs) in OA alters the expression and regulation of MMPs, and also to test the potential involvement of mitogen activated protein kinase (MAPK) signalling pathway during this process.

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The multi-criteria decision making methods, Preference METHods for Enrichment Evaluation (PROMETHEE) and Graphical Analysis for Interactive Assistance (GAIA), and the two-way Positive Matrix Factorization (PMF) receptor model were applied to airborne fine particle compositional data collected at three sites in Hong Kong during two monitoring campaigns held from November 2000 to October 2001 and November 2004 to October 2005. PROMETHEE/GAIA indicated that the three sites were worse during the later monitoring campaign, and that the order of the air quality at the sites during each campaign was: rural site > urban site > roadside site. The PMF analysis on the other hand, identified 6 common sources at all of the sites (diesel vehicle, fresh sea salt, secondary sulphate, soil, aged sea salt and oil combustion) which accounted for approximately 68.8 ± 8.7% of the fine particle mass at the sites. In addition, road dust, gasoline vehicle, biomass burning, secondary nitrate, and metal processing were identified at some of the sites. Secondary sulphate was found to be the highest contributor to the fine particle mass at the rural and urban sites with vehicle emission as a high contributor to the roadside site. The PMF results are broadly similar to those obtained in a previous analysis by PCA/APCS. However, the PMF analysis resolved more factors at each site than the PCA/APCS. In addition, the study demonstrated that combined results from multi-criteria decision making analysis and receptor modelling can provide more detailed information that can be used to formulate the scientific basis for mitigating air pollution in the region.

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Technology has provided consumers with the means to control and edit the information that they receive and share effectively, especially in the online environment. Although previous studies have investigated advertising avoidance in traditional media and on the Internet, there has been little investigation of advertising on social networking sites. This exploratory study examines the antecedents of advertising avoidance on online social networking sites, leading to the development of a model. The model suggests that advertising in the online social networking environment is more likely to be avoided if the user has expectations of a negative experience, the advertising is not relevant to the user, the user is skeptical toward the advertising message, or the consumer is skeptical toward the advertising medium.

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Heart failure is a complex disorder, characterized by activation of the sympathetic nervous system, leading to dysregulated Ca2+ homeostasis in cardiac myocytes and tissue remodeling. In a variety of diseases, cardiac malfunction is associated with aberrant fluxes of Ca2+ across both the surface membrane and the internal Ca2+ store, the sarcoplasmic reticulum (SR). One prominent hypothesis residues is that in heart failure, the activity of the ryanodine receptor (RyR2) Ca2+ release channel in the SR is increased due to excess phosphorylation and that this contributes to excess SR Ca2+ leak in diastole, reduced SR Ca2+ load and decreased contractility (Huke & Bers, 2008). There is controversy over which serine residues in RyR2 are hyperphosphorylated in animal models of heart failure and whether this is via the CaMKII or the PKA-linked signaling pathway. S2808, S2814 and S2030 in RyR2 have been variously claimed to be hyperphosphorylated. Our aim was to examine the degree of phosphorylation of these residues in RyR2 from failing human hearts. The use of human tissue was approved by the Human Research Ethics Committee, The Prince Charles Hospital, EC28114. Left ventricular tissue samples were obtained from an explanted heart of a patient with endstage heart failure (Emery Dreifuss Muscular Dystrophy with cardiomyopathy) and non-failing tissue was from a patient with cystic fibrosis undergoing heart-lung transplantation with no history of heart disease. SR vesicles were prepared as described by Laver et al. (1995) and examined with SDS-Page and Western Blot. Transferred proteins were probed with antibodies to detect total protein phosphorylation, phosphorylation of RyR2 serine residues S2808, S2814, S2030 and for the key proteins calsequestrin, triadin, junctin and FKBP12.6. To avoid membrane stripping artifact, each membrane was exposed to one phosphorylation-specific antibody and signal densities quantified using Bio-Rad Quantity One software. We found no distinguishable difference between failing and healthy hearts in the protein expression levels of RyR2, triadin, junctin or calsequestrin. We found an expected upregulation of total RyR2 phosphorylation in the failing heart sample, compared to a matched amount of RyR2 (quantified using densiometry) in healthy heart. Probing with antibodies detecting only the phosphorylated form of the specific RyR2 residues showed that the increase in total RyR2 phosphorylation in the failing heart was due to hyperphosphorylation of S2808 and S2814. We found that S2030 phosphorylation levels were unchanged in human heart failure. Interestingly, we found that S2030 has a basal level of phosphorylation in the healthy human heart, different from the absence of basal phosphorylation recently reported in rodent heart (Huke & Bers, 2008). Finally, preliminary results indicate that less FKBP 12.6 is associated with RyR2 in the failing heart, possibly as a consequence of PKA activation. In conclusion, residues S2808 and S2814 are hyperphosphorylated in human heart failure, presumably due to upregulation of the CaMKII and/or PKA signaling pathway as a result of chronic activation of the sympathetic nervous system. Such changes in RyR2 phosphorylation are believed to contribute to the leaky RyR2 phenotype associated with heart failure, which increases the incidence of arrhythmia and contributes to the severely impaired contractile performance of the failing heart. Huke S & Bers DM. (2008). Ryanodine receptor phosphorylation at serine 2030, 2808 and 2814 in rat cardiomyocytes. Biochemical and Biophysical Research Communications 376, 80-85. Laver DR, Roden LD, Ahern GP, Eager KR, Junankar PR & Dulhunty AF. (1995). Cytoplasmic Ca2+ inhibits the ryanodine receptor from cardiac muscle. Journal of Membrane Biology 147, 7-22. Proceedings

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We have recently demonstrated the geographic isolation of rice tungro bacilliform virus (RTBV) populations in the tungro-endemic provinces of Isabela and North Cotabato, Philippines. In this study, we examined the genetic structure of the virus populations at the tungro-outbreak sites of Lanao del Norte, a province adjacent to North Cotabato. We also analyzed the virus populations at the tungro-endemic sites of Subang, Indonesia, and Dien Khanh, Vietnam. Total DNA extracts from 274 isolates were digested with EcoRV restriction enzyme and hybridized with a full-length probe of RTBV. In the total population, 22 EcoRV-restricted genome profiles (genotypes) were identified. Although overlapping genotypes could be observed, the outbreak sites of Lanao del Norte had a genotype combination distinct from that of Subang or Dien Khanh but a genotype combination similar to that identified earlier from North Cotabato, the adjacent endemic province. Sequence analysis of the intergenic region and part of the ORF1 RTBV genome from randomly selected genotypes confirms the geographic clustering of RTBV genotypes and, combined with restriction analysis, the results suggest a fragmented spatial distribution of RTBV local populations in the three countries. Because RTBV depends on rice tungro spherical virus (RTSV) for transmission, the population dynamics of both tungro viruses were then examined at the endemic and outbreak sites within the Philippines. The RTBV genotypes and the coat protein RTSV genotypes were used as indicators for virus diversity. A shift in population structure of both viruses was observed at the outbreak sites with a reduced RTBV but increased RTSV gene diversity

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The use of social networking sites (SNS) by online citizens to share photos, update friends, play games and to connect with the world has exploded, with SNS and blogs now eclipsing email traffic (eMarketer 2009). Just one popular application on one SNS, (Farmville on Facebook) acquired more than 63 million users since its launch in June 2009 (Marketing 2009. The major global social networks are Facebook, Twitter, YouTube and MySpace, with Facebook claiming that it passed 350 million users in November (Marketing 2009). As usage increases and competition intensifies, the major sites must strategically position themselves to develop a competitive advantage in order to maintain or grow their share of the pie. So how do the major SNS position their brands, and do users perceive significant differences among the big players? This presentation answers these questions by reporting the results of an empirical study of SNS usage by Australian adults. Like other brands, aligning brand positioning strategies with user knowledge and perceptions of SNS is an important ingredient to achieving success (Keller 1993). Furthermore we compare the types of value for three different SNS to identify the relationships between the value derived by users and the stated positioning of the site.

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This article examines the social networking phenomenon that has been so readily embraced by school-age adolescents, in the context of its potential to contribute further to the mechanisms for and incidence of cyberbullying amongst school students. Cyberbullying in these online for a, as a misuse of technology to harass, intimidate, tease, threaten, abuse or otherwise terrorise peers, teachers and/or the school in general, is discussed from both the psychological perspective and in terms of its legal ramifications (both criminal and civil) in Australia. Some recommendations for proactive and preventative measures, education and policy adoptions are provided, together with general advice to parents, schools and adolescents on awareness of the risks involved and how young people might better protect themselves in light of that knowledge.

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Aims: This study determined whether the visibility benefits of positioning retroreflective strips in biological motion configurations were evident at real world road worker sites. ---------- Methods: 20 visually normal drivers (M=40.3 years) participated in this study that was conducted at two road work sites (one suburban and one freeway) on two separate nights. At each site, four road workers walked in place wearing one of four different clothing options: a) standard road worker night vest, b) standard night vest plus retroreflective strips on thighs, c) standard night vest plus retroreflective strips on ankles and knees, d) standard night vest plus retroreflective strips on eight moveable joints (full biomotion). Participants seated in stationary vehicles at three different distances (80m, 160m, 240m) rated the relative conspicuity of the four road workers using a series of a standardized visibility and ranking scales. ---------- Results: Adding retroreflective strips in the full biomotion configuration to the standard night vest significantly (p<0.001) enhanced perceptions of road worker visibility compared to the standard vest alone, or in combination with thigh retroreflective markings. These visibility benefits were evident at all distances and at both sites. Retroreflective markings at the ankles and knees also provided visibility benefits compared to the standard vest, however, the full biomotion configuration was significantly better than all of the other configurations. ---------- Conclusions: These data provide the first evidence that the benefits of biomotion retroreflective markings that have been previously demonstrated under laboratory and closed- and open-road conditions are also evident at real work sites.

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hSSB1 is a newly discovered single-stranded DNA (ssDNA)-binding protein that is essential for efficient DNA double-strand break signalling through ATM. However, the mechanism by which hSSB1 functions to allow efficient signalling is unknown. Here, we show that hSSB1 is recruited rapidly to sites of double-strand DNA breaks (DSBs) in all interphase cells (G1, S and G2) independently of, CtIP, MDC1 and the MRN complex (Rad50, Mre11, NBS1). However expansion of hSSB1 from the DSB site requires the function of MRN. Strikingly, silencing of hSSB1 prevents foci formation as well as recruitment of MRN to sites of DSBs and leads to a subsequent defect in resection of DSBs as evident by defective RPA and ssDNA generation. Our data suggests that hSSB1 functions upstream of MRN to promote its recruitment at DSBs and is required for efficient resection of DSBs. These findings, together with previous work establish essential roles of hSSB1 in controlling ATM activation and activity, and subsequent DSB resection and homologous recombination (HR).

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A number of instructors have recently adopted social network sites (SNSs) for learning. However, the learning design of SNSs often remains at a preliminary level similar to a personal log book because it does not properly include reflective learning elements such as individual reflection and collaboration. This article looks at the reflective learning process and the public writing process as a way of improving the quality of reflective learning on SNSs. It proposes a reflective learning model on SNSs based on two key pedagogical concepts for social networking: individual expression and collaborative connection. It is expected that the model would be helpful for instructors in designing a reflective learning process on SNSs in an effective and flexible way.

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Introduction: Osteoarthritis (OA) is the most common musculoskeletal disorder and represents a major health burden to society. In the course of the pathological development of OA, articular cartilage chondrocytes (ACCs) undergo a typical phenotype changes characterized by the expression of hypertrophic differentiation markers. Also, the adjacent subchondral bone shows signs of abnormal mineral density and enhanced production of bone turnover markers, indicative of osteoblast dysfunction. However, the mechanism(s) by which these changes occur during the OA development are not completely understood. Materials and Methods: ACCs and subchondral bone osteoblasts (SBOs) were harvested from OA and healthy patients for the cross-talk studies between normal and OA ACCs and SBOs. The involvement of mitogen activated protein kinase (MAPK) signalling pathway during the cell-cell interactions was analysed by zymography, ELISA and western blotting methods. Results: The direct and in-direct co-culture studies showed that OA (ACCs and SBOs) cells induced osteoarthritic changes of normal (ACC and SBOs) cells. This altered cell interaction induced by OA cells significantly aggravated the proteolytic activity, which resulted cartilage degeneration. The altered cell interaction appeared to significantly activate ERK 1/2 phosphorylation and inhibition of MAPK-ERK 1/2 pathway reversed the osteoarthrtitic phenotypic changes. Discussion and Conclusion: Our study has demonstrated that the altered bi-directional communication of SBOs and ACCs are critical for initiation and progression of OA related changes and that this process is mediated by MAPK signalling pathways. Targeting these altered interactions by the use of MAPK inhibitors may provide the scientific rationale for the development of novel therapeutic strategies in the treatment and management of OA related disorders.

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On obstacle-cluttered construction sites, understanding the motion characteristics of objects is important for anticipating collisions and preventing accidents. This study investigates algorithms for object identification applications that can be used by heavy equipment operators to effectively monitor congested local environment. The proposed framework contains algorithms for three-dimensional spatial modeling and image matching that are based on 3D images scanned by a high-frame rate range sensor. The preliminary results show that an occupancy grid spatial modeling algorithm can successfully build the most pertinent spatial information, and that an image matching algorithm is best able to identify which objects are in the scanned scene.

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Visual recording devices such as video cameras, CCTVs, or webcams have been broadly used to facilitate work progress or safety monitoring on construction sites. Without human intervention, however, both real-time reasoning about captured scenes and interpretation of recorded images are challenging tasks. This article presents an exploratory method for automated object identification using standard video cameras on construction sites. The proposed method supports real-time detection and classification of mobile heavy equipment and workers. The background subtraction algorithm extracts motion pixels from an image sequence, the pixels are then grouped into regions to represent moving objects, and finally the regions are identified as a certain object using classifiers. For evaluating the method, the formulated computer-aided process was implemented on actual construction sites, and promising results were obtained. This article is expected to contribute to future applications of automated monitoring systems of work zone safety or productivity.

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Australian construction and building workers are exposed to serious workplace risks - including injury, illness and death - and although there have been improvements in occupational health and safety (OHS) performance over the past 20 years, the injury and fatality rate in the Australian construction industry remains a matter of concern. The concept of safety culture is rapidly being adopted in the industry, including recognising the critical role that organisational leaders play in overall safety performance. This paper reviews recent research in construction safety leadership and provides some examples and applications relevant to risk reduction in the workforce. By focusing on developing safety competency in those that fulfil safety critical roles, and clearly articulating the relevant safety management tasks, leaders can positively influence the organisation’s safety culture. Finally, some promising research on Safety Effectiveness Indicators (SEIs) may be an industry-friendly solution to reducing workplace risks across the industry, by providing a credible, accurate, and timely measure of safety performance.