102 research outputs found

    Saltstone Osmotic Pressure

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    Recent research into the moisture retention properties of saltstone suggest that osmotic pressure may play a potentially significant role in contaminant transport (Dixon et al., 2009 and Dixon, 2011). The Savannah River Remediation Closure and Disposal Assessments Group requested the Savannah River National Laboratory (SRNL) to conduct a literature search on osmotic potential as it relates to contaminant transport and to develop a conceptual model of saltstone that incorporates osmotic potential. This report presents the findings of the literature review and presents a conceptual model for saltstone that incorporates osmotic potential. The task was requested through Task Technical Request HLW-SSF-TTR-2013-0004. Simulated saltstone typically has very low permeability (Dixon et al. 2008) and pore water that contains a large concentration of dissolved salts (Flach and Smith 2013). Pore water in simulated saltstone has a high salt concentration relative to pore water in concrete and groundwater. This contrast in salt concentration can generate high osmotic pressures if simulated saltstone has the properties of a semipermeable membrane. Estimates of osmotic pressure using results from the analysis of pore water collected from simulated saltstone show that an osmotic pressure up to 2790 psig could be generated within the saltstone. Most semi-permeable materials are non-ideal and have an osmotic efficiency <1 and as a result actual osmotic pressures are less than theoretical pressures. Observations from laboratory tests of simulated saltstone indicate that it may exhibit the behavior of a semi-permeable membrane. After several weeks of back pressure saturation in a flexible wall permeameter (FWP) the membrane containing a simulated saltstone sample appeared to have bubbles underneath it. Upon removal from the FWP the specimen was examined and it was determined that the bubbles were due to liquid that had accumulated between the membrane and the sample. One possible explanation for the accumulation of solution between the membrane and sample is the development of osmotic pressure within the sample. Osmotic pressure will affect fluid flow and contaminant transport and may result in the changes to the internal structure of the semi-permeable material. BοΏ½nard et al. 2008 reported swelling of wet cured Portland cement mortars containing salts of NaNO{sub 3}, KNO{sub 3}, Na{sub 3}PO{sub 4}x12H {sub 2}O, and K{sub 3}PO{sub 4} when exposed to a dilute solution. Typically hydraulic head is considered the only driving force for groundwater in groundwater models. If a low permeability material containing a concentrated salt solution is present in the hydrogeologic sequence large osmotic pressures may develop and lead to misinterpretation of groundwater flow and solute transport. The osmotic pressure in the semi-permeable material can significantly impact groundwater flow in the vicinity of the semi-permeable material. One possible outcome is that groundwater will flow into the semi-permeable material resulting in hydrologic containment within the membrane. Additionally, hyperfiltration can occur within semi-permeable materials when water moves through a membrane into the more concentrated solution and dissolved constituents are retained in the lower concentration solution. Groundwater flow and transport equations that incorporate chemical gradients (osmosis) have been developed. These equations are referred to as coupled flow equations. Currently groundwater modeling to assess the performance of saltstone waste forms is conducted using the PORFLOW groundwater flow and transport model. PORFLOW does not include coupled flow from chemico-osmotic gradients and therefore numerical simulation of the effect of coupled flow on contaminant transport in and around saltstone cannot be assessed. Most natural semi-permeable membranes are non-ideal membranes and do not restrict all movement of solutes and as a result theoretical osmotic potential is not realized. Osmotic efficiency is a parameter in the coupled flow equation that accounts for the non-ideal behavior of most semi-permeable membranes. On order to evaluate the effects of osmotic potential on the hydraulic of a system the osmotic potential must be known. Several lab methods have been developed to measure osmotic efficiency for use in coupled flow analysis

    Research and Development of Automated Eddy Current Testing for Composite Overwrapped Pressure Vessels

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    Eddy current testing (ET) was used to scan bare metallic liners used in the fabrication of composite overwrapped pressure vessels (COPVs) for flaws which could result in premature failure of the vessel. The main goal of the project was to make improvements in the areas of scan signal to noise ratio, sensitivity of flaw detection, and estimation of flaw dimensions. Scan settings were optimized resulting in an increased signal to noise ratio. Previously undiscovered flaw indications were observed and investigated. Threshold criteria were determined for the system software's flaw report and estimation of flaw dimensions were brought to an acceptable level of accuracy. Computer algorithms were written to import data for filtering and a numerical derivative filtering algorithm was evaluated

    AUF1/hnRNP D Represses Expression of VEGF in Macrophages

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    Vascular endothelial growth factor (VEGF) is a regulator of vascularization in development and is a key growth factor in tissue repair. In disease, VEGF contributes to vascularization of solid tumors and arthritic joints. This study examines the role of the mRNA-binding protein AUF1/heterogeneous nuclear ribonucleoprotein D (AUF1) in VEGF gene expression. We show that overexpression of AUF1 in mouse macrophage-like RAW-264.7 cells suppresses endogenous VEGF protein levels. To study 3β€² untranslated region (UTR)–mediated regulation, we introduced the 3β€² UTR of VEGF mRNA into a luciferase reporter gene. Coexpression of AUF1 represses VEGF-3β€² UTR reporter expression in RAW-264.7 cells and in mouse bone marrow–derived macrophages. The C-terminus of AUF1 contains arginine–glycine–glycine (RGG) repeat motifs that are dimethylated. Deletion of the RGG domain of AUF1 eliminated the repressive effects of AUF1. Surprisingly, expression of an AUF1-RGG peptide reduced endogenous VEGF protein levels and repressed VEGF-3β€² UTR reporter activity in RAW-264.7 cells. These findings demonstrate that AUF1 regulates VEGF expression, and this study identifies an RGG peptide that suppresses VEGF gene expression

    Use of Modal Acoustic Emission to Monitor Damage Progression in Carbon Fiber/Epoxy Tows and Implications for Composite Structures

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    This slide presentation reviews the use of Modal Acoustic Emission to monitor damage progression to carbon fiber/epoxy tows. There is a risk for catastrophic failure of composite overwrapped pressure vessels (COPVs) due to burst-before-leak (BBL) stress rupture (SR) failure of carbon-epoxy (C/Ep) COPVs. A lack of quantitative nondestructive evaluation (NDE) is causing problems in current and future spacecraft designs. It is therefore important to develop and demonstrate critical NDE that can be implemented during stages of the design process since the observed rupture can occur with little of no advanced warning. Therefore a program was required to develop quantitative acoustic emission (AE) procedures specific to C/Ep overwraps, but which also have utility for monitoring damage accumulation in composite structure in general, and to lay the groundwork for establishing critical thresholds for accumulated damage in composite structures, such as COPVs, so that precautionary or preemptive engineering steps can be implemented to minimize of obviate the risk of catastrophic failure. A computed Felicity Ratio (FR) coupled with fast Fourier Transform (FFT) frequency analysis shows promise as an analytical pass/fail criterion. The FR analysis and waveform and FFT analysis are reviewe

    Interactions between visual and semantic processing during object recognition revealed by modulatory effects of age of acquisition

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    The age of acquisition (AoA) of objects and their names is a powerful determinant of processing speed in adulthood, with early-acquired objects being recognized and named faster than late-acquired objects. Previous research using fMRI (Ellis et al., 2006. Traces of vocabulary acquisition in the brain: evidence from covert object naming. NeuroImage 33, 958–968) found that AoA modulated the strength of BOLD responses in both occipital and left anterior temporal cortex during object naming. We used magnetoencephalography (MEG) to explore in more detail the nature of the influence of AoA on activity in those two regions. Covert object naming recruited a network within the left hemisphere that is familiar from previous research, including visual, left occipito-temporal, anterior temporal and inferior frontal regions. Region of interest (ROI) analyses found that occipital cortex generated a rapid evoked response (~ 75–200 ms at 0–40 Hz) that peaked at 95 ms but was not modulated by AoA. That response was followed by a complex of later occipital responses that extended from ~ 300 to 850 ms and were stronger to early- than late-acquired items from ~ 325 to 675 ms at 10–20 Hz in the induced rather than the evoked component. Left anterior temporal cortex showed an evoked response that occurred significantly later than the first occipital response (~ 100–400 ms at 0–10 Hz with a peak at 191 ms) and was stronger to early- than late-acquired items from ~ 100 to 300 ms at 2–12 Hz. A later anterior temporal response from ~ 550 to 1050 ms at 5–20 Hz was not modulated by AoA. The results indicate that the initial analysis of object forms in visual cortex is not influenced by AoA. A fastforward sweep of activation from occipital and left anterior temporal cortex then results in stronger activation of semantic representations for early- than late-acquired objects. Top-down re-activation of occipital cortex by semantic representations is then greater for early than late acquired objects resulting in delayed modulation of the visual response

    Efficient Allele-Specific Targeting of LRRK2 R1441 Mutations Mediated by RNAi

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    Since RNA interference (RNAi) has the potential to discriminate between single nucleotide changes, there is growing interest in the use of RNAi as a promising therapeutical approach to target dominant disease-associated alleles. Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene have been linked to dominantly inherited Parkinson's disease (PD). We focused on three LRRK2 mutations (R1441G/C and the more prevalent G2109S) hoping to identify shRNAs that would both recognize and efficiently silence the mutated alleles preferentially over the wild-type alleles. Using a luciferase-based reporter system, we identified shRNAs that were able to specifically target the R1441G and R1441C alleles with 80% silencing efficiency. The same shRNAs were able to silence specifically mRNAs encoding either partial or full-length mutant LRRK2 fusion proteins, while having a minimal effect on endogenous wild-type LRRK2 expression when transfected in 293FT cells. Shifting of the mutant recognition site (MRS) from position 11 to other sites (4 and 16, within the 19-mer window of our shRNA design) reduced specificity and overall silencing efficiency. Developing an allele-specific RNAi of G2019S was problematic. Placement of the MRS at position 10 resulted in efficient silencing of reporters (75–80%), but failed to discriminate between mutant and wild-type alleles. Shifting of the MRS to positions 4, 5, 15, 16 increased the specificity of the shRNAs, but reduced the overall silencing efficiency. Consistent with previous reports, these data confirm that MRS placement influences both allele-specificity and silencing strength of shRNAs, while further modification to hairpin design or MRS position may lead to the development of effective G2019S shRNAs. In summary, the effective shRNA against LRRK2 R1441 alleles described herein suggests that RNAi-based therapy of inherited Parkinson's disease is a viable approach towards developing effective therapeutic interventions for this serious neurodegenerative disease

    Gentamicin Rapidly Inhibits Mitochondrial Metabolism in High-Frequency Cochlear Outer Hair Cells

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    Aminoglycosides (AG), including gentamicin (GM), are the most frequently used antibiotics in the world and are proposed to cause irreversible cochlear damage and hearing loss (HL) in 1/4 of the patients receiving these life-saving drugs. Akin to the results of AG ototoxicity studies, high-frequency, basal turn outer hair cells (OHCs) preferentially succumb to multiple HL pathologies while inner hair cells (IHCs) are much more resilient. To determine if endogenous differences in IHC and OHC mitochondrial metabolism dictate differential sensitivities to AG-induced HL, IHC- and OHC-specific changes in mitochondrial reduced nicotinamide adenine dinucleotide (NADH) fluorescence during acute (1 h) GM treatment were compared. GM-mediated decreases in NADH fluorescence and succinate dehydrogenase activity were observed shortly after GM application. High-frequency basal turn OHCs were found to be metabolically biased to rapidly respond to alterations in their microenvironment including GM and elevated glucose exposures. These metabolic biases may predispose high-frequency OHCs to preferentially produce cell-damaging reactive oxygen species during traumatic challenge. Noise-induced and age-related HL pathologies share key characteristics with AG ototoxicity, including preferential OHC loss and reactive oxygen species production. Data from this report highlight the need to address the role of mitochondrial metabolism in regulating AG ototoxicity and the need to illuminate how fundamental differences in IHC and OHC metabolism may dictate differences in HC fate during multiple HL pathologies

    Transcriptome dynamics and molecular cross-talk between bovine oocyte and its companion cumulus cells

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    <p>Abstract</p> <p>Background</p> <p>The bi-directional communication between the oocyte and its companion cumulus cells (CCs) is crucial for development and functions of both cell types. Transcripts that are exclusively expressed either in oocytes or CCs and molecular mechanisms affected due to removal of the communication axis between the two cell types is not investigated at a larger scale. The main objectives of this study were: 1. To identify transcripts exclusively expressed either in oocyte or CCs and 2. To identify those which are differentially expressed when the oocyte is cultured with or without its companion CCs and vice versa.</p> <p>Results</p> <p>We analyzed transcriptome profile of different oocyte and CC samples using Affymetrix GeneChip Bovine Genome array containing 23000 transcripts. Out of 13162 genes detected in germinal vesicle (GV) oocytes and their companion CCs, 1516 and 2727 are exclusively expressed in oocytes and CCs, respectively, while 8919 are expressed in both. Similarly, of 13602 genes detected in metaphase II (MII) oocytes and CCs, 1423 and 3100 are exclusively expressed in oocytes and CCs, respectively, while 9079 are expressed in both. A total of 265 transcripts are differentially expressed between oocytes cultured with (OO + CCs) and without (OO - CCs) CCs, of which 217 and 48 are over expressed in the former and the later groups, respectively. Similarly, 566 transcripts are differentially expressed when CCs mature with (CCs + OO) or without (CCs - OO) their enclosed oocytes. Of these, 320 and 246 are over expressed in CCs + OO and CCs - OO, respectively.</p> <p>While oocyte specific transcripts include those involved in transcription (<it>IRF6, POU5F1, MYF5, MED18</it>), translation (<it>EIF2AK1, EIF4ENIF1</it>) and CCs specific ones include those involved in carbohydrate metabolism (<it>HYAL1, PFKL, PYGL, MPI</it>), protein metabolic processes (<it>IHH, APOA1, PLOD1</it>), steroid biosynthetic process (<it>APOA1, CYP11A1, HSD3B1, HSD3B7</it>). Similarly, while transcripts over expressed in OO + CCs are involved in carbohydrate metabolism (<it>ACO1, 2</it>), molecular transport (<it>GAPDH, GFPT1</it>) and nucleic acid metabolism (<it>CBS, NOS2</it>), those over expressed in CCs + OO are involved in cellular growth and proliferation (<it>FOS, GADD45A</it>), cell cycle (<it>HAS2, VEGFA</it>), cellular development (<it>AMD1, AURKA, DPP4</it>) and gene expression (<it>FOSB, TGFB2</it>).</p> <p>Conclusion</p> <p>In conclusion, this study has generated large scale gene expression data from different oocyte and CCs samples that would provide insights into gene functions and interactions within and across different pathways that are involved in the maturation of bovine oocytes. Moreover, the presence or absence of oocyte and CC factors during bovine oocyte maturation can have a profound effect on transcript abundance of each cell types, thereby showing the prevailing molecular cross-talk between oocytes and their corresponding CCs.</p
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