Results are presented on R&D efforts to design and build large size veto panels, optimized for underground low background experiments, in the most efficient and economical way using commercially ...available components. A variety of plastic scintillators, photomultiplier tubes, wavelength shifting fibers, and light reflector combinations were tested. Results of these studies and performance of a 2.2m long panel are presented.
Aim To assess whether there is a significant difference in the incidence of patients with metastases of rectal carcinoma at 1 year follow-up between magnetic resonance imaging (MRI)-detected ...extramural venous invasion (EMVI) and those without. Materials and methods A search of our institution's cancer registry revealed 788 patients with rectal carcinoma between January 2007 and April 2012. Those who were initially staged using MRI and computed tomography (CT) chest/abdomen/pelvis, and followed-up with a CT chest/abdomen/pelvis examination at 1 year were included in this retrospective study. Patients with synchronous metastases were excluded, leaving a cohort of 202 cases. Two consultant radiologists reviewed all MRI images and gave a consensus opinion regarding EMVI grade and vessel size involved. All CT images were reviewed for metastases. Results were analysed using chi-squared and Fisher's exact tests. Results There were 53 cases (26.2%) of EMVI-positive rectal carcinoma. Of the patients with EMVI, 24.5% developed metastases at 1 year follow-up, compared to 6.7% of those without. There is a significant difference in prognosis between those patients with and those without MRI-detected EMVI (χ2 = 12.29, p < 0·001). Those with EMVI have a 3.7 times increased relative risk of developing metachronous metastases within 1 year of diagnosis. Conclusion MRI-detected EMVI-positive rectal carcinomas are associated with an increased risk of metachronous metastases within 1 year of diagnosis. Currently, EMVI status does not directly influence the initial management of rectal carcinoma. This available and potentially prognostic feature could be used to guide treatment pathways to increase disease-free survival.
Misfolding and aggregation of huntingtin is one of the hallmarks of Huntington disease, but the overall structure of these aggregates and the mechanisms by which huntingtin misfolds remain poorly ...understood. Here we used site-directed spin labeling and electron paramagnetic resonance (EPR) spectroscopy to study the structural features of huntingtin exon 1 (HDx1) containing 46 glutamine residues in its polyglutamine (polyQ) region. Despite some residual structuring in the N terminus, we find that soluble HDx1 is highly dynamic. Upon aggregation, the polyQ domain becomes strongly immobilized indicating significant tertiary or quaternary packing interactions. Analysis of spin-spin interactions does not show the close contact between same residues that is characteristic of the parallel, in-register structure commonly found in amyloids. Nevertheless, the same residues are still within 20 Å of each other, suggesting that polyQ domains from different molecules come into proximity in the fibrils. The N terminus has previously been found to take up a helical structure in fibrils. We find that this domain not only becomes structured, but that it also engages in tertiary or quaternary packing interactions. The existence of spin-spin interactions in this region suggests that such contacts could be made between N-terminal domains from different molecules. In contrast, the C-terminal domain is dynamic, contains polyproline II structure, and lacks pronounced packing interactions. This region must be facing away from the core of the fibrils. Collectively, these data provide new constraints for building structural models of HDx1 fibrils.
Background: The structure of fibrils formed in Huntington disease remains unknown.
Results: Local structure and domain organization of huntingtin exon 1 fibrils were determined by EPR spectroscopy.
Conclusion: In contrast to the C terminus, the N terminus, and polyglutamine core regions become closely packed, without forming a parallel, in-register structure.
Significance:The results provide structural constraints and insight into how adjacent domains affect polyglutamine structure.
In comparison to most modern teleost fishes, sturgeons generally display muted stress responses. While a muted stress response appears to be ubiquitous across sturgeon species, the mechanisms ...unpinning this muted response have not been fully described. The objective of this study was to determine the patterns of hematological and transcriptomic change in muscle tissue following an acute high temperature stress (critical thermal maxima; CTmax) in two locally co-occurring but evolutionarily distant sturgeon species (Atlantic and shortnose sturgeon). The most striking pattern found was that Atlantic sturgeon launched a vigorous transcriptomic response at CTmax, whereas shortnose sturgeon did not. In contrast, shortnose sturgeon have significantly higher cortisol than Atlantics at CTmax, reconfirming that shortnose have a less muted cortisol stress response. Atlantic sturgeon downregulated a number of processes, included RNA creation/processing, methylation and immune processes. Furthermore, a number of genes related to heat shock proteins were differentially expressed at CTmax in Atlantic sturgeon but none of these genes were significantly changed in shortnose sturgeon. We also note that the majority of differentially expressed genes of both species are undescribed and have no known orthologues. These results suggest that, while sturgeons as a whole may show muted stress responses, individual sturgeon species likely use different inducible strategies to cope with acute high temperature stress.
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•Atlantic and shortnose sturgeon likely utilize divergent inducible responses to acute thermal stress.•Atlantic sturgeon launch a robust transcriptomic response to acute heat stress yet shortnose sturgeon do not.•Genes related to heat shock protein are differentially expressed at CTmax by Atlantic but not in shortnose sturgeon.•Contrarily, acute heat stress elicits a higher cortisol response in shortnose compared to Atlantic sturgeon.•RNA processing, transcription and methylation was downregulated in Atlantic sturgeon.
Patients with lower extremity peripheral artery disease (PAD) have decreased mobility, which is not fully explained by impaired blood supply to the lower limb. Additionally, reports are conflicted ...regarding fiber type distribution patterns in PAD, but agree that skeletal muscle mitochondrial respiration is impaired.
To test the hypothesis that reduced muscle fiber oxidative activity and type I distribution are negatively associated with walking performance in PAD, calf muscle biopsies from non-PAD (n = 7) and PAD participants (n = 26) were analyzed immunohistochemically for fiber type and size, oxidative activity, markers of autophagy, and capillary density. Data were analyzed using analysis of covariance.
There was a wide range in fiber type distribution among subjects with PAD (9-81 % type I fibers) that did not correlate with walking performance. However, mean type I fiber size correlated with 4-min normal- and fastest-paced walk velocity (r = 0.4940, P = 0.010 and r = 0.4944, P = 0.010, respectively). Although intensity of succinate dehydrogenase activity staining was consistent with fiber type, up to 17 % of oxidative fibers were devoid of mitochondria in their cores, and the core showed accumulation of the autophagic marker, LC3, which did not completely co-localize with LAMP2, a lysosome marker.
Calf muscle type I fiber size positively correlates with walking performance in PAD. Accumulation of LC3 and a lack of co-localization of LC3 with LAMP2 in the area depleted of mitochondria in PAD fibers suggests impaired clearance of damaged mitochondria, which may contribute to reduced muscle oxidative capacity. Further study is needed to determine whether defective mitophagy is associated with decline in function over time, and whether interventions aimed at preserving mitochondrial function and improving autophagy can improve walking performance in PAD.
Although expanded polyglutamine (polyQ) repeats are inherently toxic, causing at least nine neurodegenerative diseases, the protein context determines which neurons are affected. The polyQ expansion ...that causes Huntington's disease (HD) is in the first exon (HDx-1) of huntingtin (Htt). However, other parts of the protein, including the 17 N-terminal amino acids and two proline (polyP) repeat domains, regulate the toxicity of mutant Htt. The role of the P-rich domain that is flanked by the polyP domains has not been explored. Using highly specific intracellular antibodies (intrabodies), we tested various epitopes for their roles in HDx-1 toxicity, aggregation, localization, and turnover. Three domains in the P-rich region (PRR) of HDx-1 are defined by intrabodies: MW7 binds the two polyP domains, and Happ1 and Happ3, two new intrabodies, bind the unique, P-rich epitope located between the two polyP epitopes. We find that the PRR-binding intrabodies, as well as V(L)12.3, which binds the N-terminal 17 aa, decrease the toxicity and aggregation of HDx-1, but they do so by different mechanisms. The PRR-binding intrabodies have no effect on Htt localization, but they cause a significant increase in the turnover rate of mutant Htt, which V(L)12.3 does not change. In contrast, expression of V(L)12.3 increases nuclear Htt. We propose that the PRR of mutant Htt regulates its stability, and that compromising this pathogenic epitope by intrabody binding represents a novel therapeutic strategy for treating HD. We also note that intrabody binding represents a powerful tool for determining the function of protein epitopes in living cells.
This book argues that the British government's repression of the 1790s rivals the French Revolution as the most important historical event for our understanding the development of Romantic ...literature. Romanticism has long been associated with both rebellion and escapism, and much Romantic historicism traces an arc from the outburst of democratic energy in British culture triggered by the French Revolution to a dwindling of enthusiasm later in the 1790s, when things in France turned violent. Writers such as Wordsworth and Coleridge can then be seen as "apostates" who turned from radical politics to a poetics of transcendence. Bugg argues instead for a poetics of silence, and his book is set against the backdrop of the so-called Gagging Acts and other legislation of William Pitt, which in literature manifests itself stylistically as silence, stuttering, fragmentation, and encoding. Mining archives of unpublished documents, including manuscripts, diaries, and letters, where authors were more candid, as well as rereading the work of both major and minor figures, a number of whom were subject to prison sentences,Five Long Winters offers a new way of approaching the literature of the Romantic era.
Proteolytic processing of mutant huntingtin (mHtt), the protein that causes Huntington's disease (HD), is critical for mHtt toxicity and disease progression. mHtt contains several caspase and calpain ...cleavage sites that generate N-terminal fragments that are more toxic than full-length mHtt. Further processing is then required for the degradation of these fragments, which in turn, reduces toxicity. This unknown, secondary degradative process represents a promising therapeutic target for HD.
We have used intrabodies, intracellularly expressed antibody fragments, to gain insight into the mechanism of mutant huntingtin exon 1 (mHDx-1) clearance. Happ1, an intrabody recognizing the proline-rich region of mHDx-1, reduces the level of soluble mHDx-1 by increasing clearance. While proteasome and macroautophagy inhibitors reduce turnover of mHDx-1, Happ1 is still able to reduce mHDx-1 under these conditions, indicating Happ1-accelerated mHDx-1 clearance does not rely on these processes. In contrast, a calpain inhibitor or an inhibitor of lysosomal pH block Happ1-mediated acceleration of mHDx-1 clearance. These results suggest that mHDx-1 is cleaved by calpain, likely followed by lysosomal degradation and this process regulates the turnover rate of mHDx-1. Sequence analysis identifies amino acid (AA) 15 as a potential calpain cleavage site. Calpain cleavage of recombinant mHDx-1 in vitro yields fragments of sizes corresponding to this prediction. Moreover, when the site is blocked by binding of another intrabody, V(L)12.3, turnover of soluble mHDx-1 in living cells is blocked.
These results indicate that calpain-mediated removal of the 15 N-terminal AAs is required for the degradation of mHDx-1, a finding that may have therapeutic implications.
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