952 research outputs found

    The expression and function of midkine in the vertebrate retina

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    Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/102704/1/bph12495.pd

    Intermitochondrial cement (IMC) harbors piRNA biogenesis machinery and exonuclease domain-containing proteins EXD1 and EXD2 in mouse spermatocytes

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    Background Germ granules are large cytoplasmic ribonucleoprotein complexes that emerge in the germline to participate in RNA regulation. The two most prominent germ granules are the intermitochondrial cement (IMC) in meiotic spermatocytes and the chromatoid body (CB) in haploid round spermatids, both functionally linked to the PIWI-interacting RNA (piRNA) pathway. Aims In this study, we clarified the IMC function by identifying proteins that form complexes with a well-known IMC protein PIWIL2/MILI in the mouse testis. Results The PIWIL2 interactome included several proteins with known functions in piRNA biogenesis. We further characterized the expression and localization of two of the identified proteins, Exonuclease 3′–5′ domain-containing proteins EXD1 and EXD2, and confirmed their localization to the IMC. We showed that EXD2 interacts with PIWIL2, and that the mutation of Exd2 exonuclease domain in mice induces misregulation of piRNA levels originating from specific pachytene piRNA clusters, but does not disrupt male fertility. Conclusion Altogether, this study highlights the central role of the IMC as a platform for piRNA biogenesis, and suggests that EXD1 and EXD2 function in the IMC-mediated RNA regulation in postnatal male germ cells.We would like to thank all Kotaja lab members for their support and help. The Turku BioScience Cell Imaging and Proteomics Core Facilities are acknowledged for their services and Turku Center for Disease Modeling (TCDM) and Turku Central Animal Facility for providing expertise on animal experimentation. This study was supported by the Academy of Finland, the Sigrid Jusélius Foundation, the Novo Nordisk Foundation, the Jalmari and Rauha Ahokas Foundation, Turku Doctoral Programme of Molecular Medicine (TuDMM), and the Finnish Cultural Foundation

    Technology Learning Curves for Energy Policy Support

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    The European Commission's Joint Research Centre and the Energy Research Centre of the Netherlands (ECN) organised an expert workshop on 'Learning Curves for Policy Support' in Amsterdam on 8 March 2012. It aimed to assess the challenges in the application of the two-factor learning curve, or alternative solutions in supporting policy decision making in the framework of the European Strategic Energy Technology Plan, and explored options for improvement. The workshop gathered distinguished experts in the field of scientific research on learning curves and policy researchers from the European Commission and ECN to assess the challenges in the application of the two-factor-learning curve, or alternative solutions in supporting policy decision making, and to provide options for improvement. This paper forms the summary of outcomes from the workshop. Due to the very different nature of the One-Factor-Learning concept and the Two-Factor-Learning concept, these are discussed in separate parts. In each of these parts the context and the methodology are introduced, methodological and data challenges are described and the problems associated with the application of the concept in models is discussed.JRC.F.6-Energy systems evaluatio

    Fabrication of Silver Nanoparticles Dispersed in Palm Oil Using Laser Ablation

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    In this study we used a laser ablation technique for preparation of silver nanoparticles. The fabrication process was carried out by ablation of a silver plate immersed in palm oil. A pulsed Nd:YAG laser at a wavelength of 1064 nm was used for ablation of the plate at different times. The palm coconut oil allowed formation of nanoparticles with very small and uniform particle size, which are dispersed very homogeneously within the solution. The obtained particle sizes for 15 and 30 minute ablation times were 2.5 and 2 nm, respectively. Stability study shows that all of the samples remained stable for a reasonable period of time

    Ejaculate Economics: Testing the Effects of Male Sexual History on the Trade-Off between Sperm and Immune Function in Australian Crickets

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    Trade-offs between investment into male sexual traits and immune function provide the foundation for some of the most prominent models of sexual selection. Post-copulatory sexual selection on the male ejaculate is intense, and therefore trade-offs should occur between investment into the ejaculate and the immune system. Examples of such trade-offs exist, including that between sperm quality and immunity in the Australian cricket, Teleogryllus oceanicus. Here, we explore the dynamics of this trade-off, examining the effects that increased levels of sexual interaction have on the viability of a male's sperm across time, and the concomitant effects on immune function. Males were assigned to a treatment, whereby they cohabited with females that were sexually immature, sexually mature but incapable of copulation, or sexually mature and capable of copulation. Sperm viability of each male was then assessed at two time points: six and 13 days into the treatment, and immune function at day 13. Sperm viability decreased across the time points, but only for males exposed to treatment classes involving sexually mature females. This decrease was similar in magnitude across both sexually mature classes, indicating that costs to the expression of high sperm viability are incurred largely through levels of pre-copulatory investment. Males exposed to immature females produced sperm of low viability at both time points. Although we confirmed a weak negative association between sperm viability and lytic activity (a measure of immune response to bacterial infection) at day 13, this relationship was not altered across the mating treatment. Our results highlight that sperm viability is a labile trait, costly to produce, and subject to strategic allocation in these crickets

    Analysis of the extreme diversity of salivary alpha-amylase isoforms generated by physiological proteolysis using liquid chromatography-tandem mass spectrometry

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    Saliva is a crucial biofluid for oral health and is also of increasing importance as a non-invasive source of disease biomarkers. Salivary alpha-amylase is an abundant protein in saliva, and changes in amylase expression have been previously associated with a variety of diseases and conditions. Salivary alpha-amylase is subject to a high diversity of post-translational modifications, including physiological proteolysis in the oral cavity. Here we developed methodology for rapid sample preparation and non-targeted LC-ESI-MS/MS analysis of saliva from healthy subjects and observed an extreme diversity of alpha-amylase proteolytic isoforms. Our results emphasize the importance of consideration of post-translational events such as proteolysis in proteomic studies, biomarker discovery and validation, particularly in saliva

    RNA Oxidation Adducts 8-OHG and 8-OHA Change with Aβ42 Levels in Late-Stage Alzheimer's Disease

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    While research supports amyloid-β (Aβ) as the etiologic agent of Alzheimer's disease (AD), the mechanism of action remains unclear. Evidence indicates that adducts of RNA caused by oxidation also represent an early phenomenon in AD. It is currently unknown what type of influence these two observations have on each other, if any. We quantified five RNA adducts by gas chromatography/mass spectroscopy across five brain regions from AD cases and age-matched controls. We then used a reductive directed analysis to compare the RNA adducts to common indices of AD neuropathology and various pools of Aβ. Using data from four disease-affected brain regions (Brodmann's Area 9, hippocampus, inferior parietal lobule, and the superior and middle temporal gyri), we found that the RNA adduct 8-hydroxyguanine (8-OHG) decreased, while 8-hydroxyadenine (8-OHA) increased in AD. The cerebellum, which is generally spared in AD, did not show disease related changes, and no RNA adducts correlated with the number of plaques or tangles. Multiple regression analysis revealed that SDS-soluble Aβ42 was the best predictor of changes in 8-OHG, while formic acid-soluble Aβ42 was the best predictor of changes in 8-OHA. This study indicates that although there is a connection between AD related neuropathology and RNA oxidation, this relationship is not straightforward
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