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131.
Christopher I. Richards Rahul Srinivasan Cheng Xiao Elisha D. W. Mackey Julie M. Miwa Henry A. Lester 《The Journal of biological chemistry》2011,286(36):31241-31249
We employed a pH-sensitive GFP analog, superecliptic phluorin, to observe aspects of nicotinic acetylcholine receptor (nAChR) trafficking to the plasma membrane (PM) in cultured mouse cortical neurons. The experiments exploit differences in the pH among endoplasmic reticulum (ER), trafficking vesicles, and the extracellular solution. The data confirm that few α4β4 nAChRs, but many α4β2 nAChRs, remain in neutral intracellular compartments, mostly the ER. We observed fusion events between nAChR-containing vesicles and PM; these could be quantified in the dendritic processes. We also studied the β4R348C polymorphism, linked to amyotrophic lateral sclerosis (ALS). This mutation depressed fusion rates of α4β4 receptor-containing vesicles with the PM by ∼2-fold, with only a small decrease in the number of nAChRs per vesicle. The mutation also decreased the number of ER exit sites, showing that the reduced receptor insertion results from a change at an early stage in trafficking. We confirm the previous report that the mutation leads to reduced agonist-induced currents; in the cortical neurons studied, the reduction amounts to 2–3-fold. Therefore, the reduced agonist-induced currents are caused by the reduced number of α4β4-containing vesicles reaching the membrane. Chronic nicotine exposure (0.2 μm) did not alter the PM insertion frequency or trafficking behavior of α4β4-laden vesicles. In contrast, chronic nicotine substantially increased the number of α4β2-containing vesicle fusions at the PM; this stage in α4β2 nAChR up-regulation is presumably downstream from increased ER exit. Superecliptic phluorin provides a tool to monitor trafficking dynamics of nAChRs in disease and addiction. 相似文献
132.
Stevens EL Heckenberg G Roberson ED Baugher JD Downey TJ Pevsner J 《PLoS genetics》2011,7(9):e1002287
It is an assumption of large, population-based datasets that samples are annotated accurately whether they correspond to known relationships or unrelated individuals. These annotations are key for a broad range of genetics applications. While many methods are available to assess relatedness that involve estimates of identity-by-descent (IBD) and/or identity-by-state (IBS) allele-sharing proportions, we developed a novel approach that estimates IBD0, 1, and 2 based on observed IBS within windows. When combined with genome-wide IBS information, it provides an intuitive and practical graphical approach with the capacity to analyze datasets with thousands of samples without prior information about relatedness between individuals or haplotypes. We applied the method to a commonly used Human Variation Panel consisting of 400 nominally unrelated individuals. Surprisingly, we identified identical, parent-child, and full-sibling relationships and reconstructed pedigrees. In two instances non-sibling pairs of individuals in these pedigrees had unexpected IBD2 levels, as well as multiple regions of homozygosity, implying inbreeding. This combined method allowed us to distinguish related individuals from those having atypical heterozygosity rates and determine which individuals were outliers with respect to their designated population. Additionally, it becomes increasingly difficult to identify distant relatedness using genome-wide IBS methods alone. However, our IBD method further identified distant relatedness between individuals within populations, supported by the presence of megabase-scale regions lacking IBS0 across individual chromosomes. We benchmarked our approach against the hidden Markov model of a leading software package (PLINK), showing improved calling of distantly related individuals, and we validated it using a known pedigree from a clinical study. The application of this approach could improve genome-wide association, linkage, heterozygosity, and other population genomics studies that rely on SNP genotype data. 相似文献
133.
Elisha P. Renne 《Visual Anthropology: Published in cooperation with the Commission on Visual Anthropology》2013,26(2):139-155
In the past seventy years the food‐spread, Vegemite, has become a symbol of Australian national identity. This paper examines the historical process by which this identity has been constituted, focussing on advertisements of Vegemite in print and on television from approximately 1923 to 1960. The marketing of Vegemite during this period coincided both with the desire for a distinctive national identity and with technological developments in food processing, communications, and health care. The promoters of these images and messages, drawing upon an egalitarian ideal of good health for all and the belief in the benefits of science, have also emphasized Vegemite's contribution to the health of Australian children. These early childhood associations with Vegemite and school suggest how the everyday intimacies of food contribute to a sense of a national identity. 相似文献
134.
Catalase activity increased in Peridinium gatunense (formerly P. cinctum fa. westii) cells during the decline of the seasonal spring bloom period in Lake Kinneret. This was correlated with the low ambient total CO2 concentration. The relationship was confirmed in laboratory experiments where maximum catalase activity occurred under an atmosphere composed of 30% O2 and 0.003% CO2. Conversely, high CO2 concentrations inhibited catalase activity. The rise in catalase activity was not directly due to increasing environmental pH, as in vitro and in vivo measurements showed a characteristic broad pH curve with a constant activity from pH 6–10 for catalase. Photoinhibition of catalase occurred above 250 μmol photons · m?2· s?1. However, at high photoinactivating irradiances, photoinhibition was ameliorated under high pO2/pCO2. Such conditions prevail in the Kinneret at the end of the spring. We propose that the enhancement of photorespiration (under high pO2/pCO2) induces a temporary burst in catalase activity despite the progressively photoinhibitory conditions of early summer. 相似文献
135.
Elisha P. Renne 《The journal of the Royal Anthropological Institute》2002,8(3):551-569
This article examines the place of religion in the process of 'becoming modern', associated with conversion to Christianity and with literacy, in one Ekiti Yoruba town in southwestern Nigeria. It questions theories of modernization that presume a shift from communal 'traditional' to private, secularized religious practice, focusing on beliefs about the genesis of life, specifically cosmology and fertility. This approach provides a means for examining assumptions about the separation of the secular and religious, evidenced through an examination of local interpretations of biblical texts associated with barrenness and extraordinary births. How these stories have been interpreted by Ekiti Yoruba women and men offers a perspective on the processes whereby forms of 'modern hybrids' are constructed, altered, and reconstructed, as well as on how the fundamental bases of fertility are understood in particular social contexts. 相似文献
136.
137.
We report the encapsulation of MIN6 cells, a pancreatic beta-cell line, using thermally induced gelable materials. This strategy uses aqueous solvent and mild temperatures during encapsulation, thereby minimizing adverse effects on cell function and viability. Using a 2:1 mixture of PNIPAAm-PEG-PNIPAAm tri-block copolymer and PNIPAAm homopolymer that exhibit reversible sol-to-gel transition at approximately 30 degrees C, gels were formed that exhibit mechanical integrity, and are stable in H(2)O, PBS and complete DMEM with negligible mass loss at 37 degrees C for 60 days. MTT assays showed undetectable cytotoxicity of the polymers towards MIN6 cells. A simple microencapsulation process was developed using vertical co-extrusion and a 37 degrees C capsule collection bath containing a paraffin layer above DMEM. Spherical capsules with diameters ranging from 500 to 900 microm were formed. SEM images of freeze-dried capsules with PBS as the core solution showed homogenous gel capsule membranes. Confocal microscopy revealed that the encapsulated cells tended to form small aggregates over 5 days, and staining for live and dead cells showed high viability post-encapsulation. A static glucose challenge with day-5 cultured microencapsulated cells exhibited glucose-dependent insulin secretion comparable to controls of free MIN6 cells grown in monolayers. These results demonstrate the potential use of these thermo-responsive polymers as cell encapsulation membranes. 相似文献
138.
Shaffer Christopher A. Milstein Marissa S. Lindsey Laramie L. Wolf Tiffany M. Suse Philip Marawanaru Elisha Kipp Evan J. Garwood Tyler Travis Dominic A. Terio Karen A. Larsen Peter A. 《International journal of primatology》2022,43(2):253-272
International Journal of Primatology - Zoonotic disease risk is greatly influenced by cultural practices and belief systems. Yet, few studies have investigated how different ways of knowing are... 相似文献
139.
Ian R Kelsall Elisha H McCrory Yingqi Xu Cheryl L Scudamore Sambit K Nanda Paula ManceboGamella Nicola T Wood Axel Knebel Stephen J Matthews Philip Cohen 《The EMBO journal》2022,41(8)
HOIL‐1, a component of the linear ubiquitin chain assembly complex (LUBAC), ubiquitylates serine and threonine residues in proteins by esterification. Here, we report that mice expressing an E3 ligase‐inactive HOIL‐1[C458S] mutant accumulate polyglucosan in brain, heart and other organs, indicating that HOIL‐1’s E3 ligase activity is essential to prevent these toxic polysaccharide deposits from accumulating. We found that HOIL‐1 monoubiquitylates glycogen and α1:4‐linked maltoheptaose in vitro and identify the C6 hydroxyl moiety of glucose as the site of ester‐linked ubiquitylation. The monoubiquitylation of maltoheptaose was accelerated > 100‐fold by the interaction of Met1‐linked or Lys63‐linked ubiquitin oligomers with the RBR domain of HOIL‐1. HOIL‐1 also transferred pre‐formed ubiquitin oligomers to maltoheptaose en bloc, producing polyubiquitylated maltoheptaose in one catalytic step. The Sharpin and HOIP components of LUBAC, but not HOIL‐1, bound to unbranched and infrequently branched glucose polymers in vitro, but not to highly branched mammalian glycogen, suggesting a potential function in targeting HOIL‐1 to unbranched glucosaccharides in cells. We suggest that monoubiquitylation of unbranched glucosaccharides may initiate their removal from cells, preventing precipitation as polyglucosan. 相似文献
140.
Carbon-13 NMR Studies of Salt Shock-Induced Carbohydrate Turnover in the Marine Cyanobacterium Agmenellum quadruplicatum 总被引:1,自引:0,他引:1
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Carbon turnover in response to abrupt changes in salinity, including the mobilization of glycogen for use in osmoregulation was studied with pulse-chase strategies utilizing nuclear magnetic resonance (NMR)-silent and NMR-detectable 12C and 13C isotopes, respectively. Growth of Agmenellum quadruplicatum in 30%-enriched13C bicarbonate provided sufficient NMR-detectability of intracellular organic osmoregulants for these studies. A comparison of NMR spectra of intact cells and their ethanol extracts showed that the intact cell data were suitable for quantitative work, and, when combined with ESR measurements of cell volumes, yielded intracellular glucosylglycerol concentrations without disrupting the cells. NMR pulse-chase experiments were used to show that 13C-enriched glycogen, which had previously been accumulated by the cells under nitrogen-limited growth at low salinities, could be utilized for the synthesis of glucosylglycerol when the cells were abruptly transferred to hypersaline media, but only in the light. It was also shown that the accumulation of glucosylglycerol in the light occurred on a time scale similar to that of cell doubling. Depletion of glucosylglycerol when cells abruptly transferred to lower salinities appeared to be rapid—the intracellular pool of this osmoregulant was decreased 2-fold within 2 hours of hypotonic shock. 相似文献