583 research outputs found
Positron annihilation studies of the AlOₓ/SiO₂/Si interface in solar cell structures
Film and film/substrate interfacecharacteristics of 30 and 60 nm-thick AlOₓfilmsgrown on Si substrates by thermal atomic layer deposition(ALD), and 30 nm-thick AlOₓfilms by sputtering, have been probed using variable-energy positron annihilation spectroscopy (VEPAS) and Doppler-broadened spectra ratio curves. All samples were found to have an interface which traps positrons, with annealing increasing this trapping response, regardless of growth method. Thermal ALD creates an AlOₓ/SiOₓ/Si interface with positron trapping and annihilation occurring in the Si side of the SiOₓ/Si boundary. An induced positive charge in the Si next to the interface reduces diffusion into the oxides and increases annihilation in the Si. In this region there is a divacancy-type response (20 ± 2%) before annealing which is increased to 47 ± 2% after annealing.Sputtering seems to not produce samples with this same electrostatic shielding; instead, positron trapping occurs directly in the SiOₓinterface in the as-deposited sample, and the positron response to it increases after annealing as an SiO₂ layer is formed. Annealing the film has the effect of lowering the film oxygen response in all film types. Compared to other structural characterization techniques, VEPAS shows larger sensitivity to differences in film preparation method and between as-deposited and annealed samples.T-T.A.L., S.R., and A.C. acknowledge funding from the
Australian Research Council
AFM imaging reveals the assembly of a P2X receptor complex containing P2X2, P2X4 and P2X6 subunits
Seven P2X purinergic receptor subunits have been identified: P2X1-P2X7. All except P2X6 assemble as homotrimers, and six heteromeric receptors (P2X1/2, P2X1/4, P2X1/5, P2X2/3, P2X2/6 and P2X4/6) have been described. In addition, P2X4 homomers associate with P2X2 or P2X7 homomers as dimers of trimers. The various P2X receptors show individual functional properties, suggesting distinct physiological roles. The overlapping expression of P2X2, P2X4 and P2X6 subunits has been shown in different cell types, and functional analysis of P2X receptors in Leydig cells suggests that the three subunits interact
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Ultra-flat Gold QCM Electrodes Fabricated with Pressure Forming Template Stripping for Protein Studies at the Nanoscale
Single-molecule imaging of proteins using atomic force microscopy (AFM) is crucially dependent on protein attachment to ultra-flat substrates. The technique of template stripping (TS), which can be used to create large areas of atomically flat gold, has been used to great effect
for this purpose. However, this approach requires an epoxy which can swell in solution, causing surface roughening and substantially increasing the thickness of any sample, preventing its use on acoustic resonators in liquid. Diffusion bonding techniques should circumvent this problem
but cannot be used on samples containing patterned features with mismatched heights due to cracking and poor transfer. Here, we describe a new technique called pressure forming template stripping (PTS) which permits an ultra-flat (0.35 ± 0.05 nm root-mean-square roughness) layer
of gold to be transferred to the surface of a patterned substrate at low temperature and pressure. We demonstrate this technique by modifying a quartz crystal microbalance (QCM) sensor to contain an ultra-flat gold surface. Standard QCM chips have substantial roughness, preventing
AFM imaging of proteins on the surface after measurement. With our approach there is no need to run samples in parallel: the modified QCM chip is flat enough to permit high-contrast AFM imaging after adsorption studies have been conducted. The PTS-QCM chips are then used to
demonstrate adsorption of bovine serum albumin in comparison to rough QCM chips. The ability to attach thin layers of ultra-flat metals to surfaces of heterogeneous nature without epoxy will have many applications in diverse fields where there is a requirement to observe nanoscale phenomena with multiple techniques, including surface and interfacial science, optics, and biosensing
Association of depression and anxiety with clinical, sociodemographic, lifestyle and environmental factors in South Asian and white European people at high risk of diabetes
AIM: To investigate the prevalence and correlates of depressive and anxiety symptoms within South Asian and white European populations at high risk of developing Type 2 diabetes. METHODS: Data were collected at baseline, and at 12, 24 and 36 months from 1429 white European people (age 64±7 years, 35.8% women) and 160 South Asian people (age 59±9 years, 30.6% women) who were at high risk of Type 2 diabetes and who took part in two Type 2 diabetes prevention trials in Leicestershire, UK. The Hospital Anxiety and Depression Scale was administered during each study visit. Clinical, sociodemographic, lifestyle and environmental data were collected. RESULTS: At baseline, the burden of depressive symptoms varied by ethnic group and gender, with 9.9% of white European men, 14.9% of white European women, 23.6% of South Asian men and 29.2% of South Asian women exceeding the cut-off score for mild-to-severe depression. During the course of the study and after adjustment for clinical, sociodemographic, lifestyle and environmental factors, depressive symptoms remained higher in the South Asian compared to the white European participants [score higher by 1.5, 95% CI 0.9-2.1]. Levels of anxiety were also higher in the South Asian participants, although associations were attenuated after adjustment. Social deprivation, BMI, proximity to fast-food outlets and physical activity were correlates for depression in both the South Asian and white European participants. CONCLUSIONS: A higher burden of depressive symptoms was consistently evident among the South Asian participants, even after adjustment for multiple covariates. It is important to understand both the reasons why these differences are present, to help reduce health inequalities, and whether higher levels of depressive symptoms affect the uptake of and retention rates in diabetes prevention programmes in South Asian communities. This article is protected by copyright. All rights reserved
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Syncollin is an antibacterial polypeptide
Funder: AstraZeneca; Id: http://dx.doi.org/10.13039/100004325Funder: David James Studentship, Department of Pharmacology, University of CambridgeAbstract: Syncollin is a 16‐kDa protein found predominantly in the zymogen granules of pancreatic acinar cells, with expression at lower levels in intestinal epithelial cells and neutrophils. Here, we used Strep‐tagged syncollin isolated from the supernatant of transiently transfected mammalian cells to test the hypothesis that syncollin has antibacterial properties, which might enable it to play a role in host defence in the gut and possibly elsewhere. We show that syncollin is an exceptionally thermostable protein with a circular dichroism spectrum consistent with a predominantly beta‐sheet structure. Syncollin binds to bacterial peptidoglycan and restricts the growth of representative Gram‐positive (Lactococcus lactis) and Gram‐negative (Escherichia coli) bacteria. Syncollin induces propidium iodide uptake into E. coli (but not L. lactis), indicating permeabilisation of the bacterial membrane. It also causes surface structural damage in both L. lactis and E. coli, as visualised by scanning electron microscopy. We propose that syncollin is a previously unidentified member of a large group of antimicrobial polypeptides that control the gut microbiome. Take Aways: Syncollin is a 16‐kDa protein found in pancreatic zymogen granules. Syncollin is highly thermostable and has a predominantly beta‐sheet structure. Syncollin binds peptidoglycan and restricts the growth of L. lactis and E. coli. Syncollin causes propidium iodide uptake into E. coli (but not L. lactis). Syncollin causes surface structural damage in both L. lactis and E. coli
Unusual structures are present in DNA fragments containing super-long Huntingtin CAG repeats.
BACKGROUND: In the R6/2 mouse model of Huntington's disease (HD), expansion of the CAG trinucleotide repeat length beyond about 300 repeats induces a novel phenotype associated with a reduction in transcription of the transgene. METHODOLOGY/PRINCIPAL FINDINGS: We analysed the structure of polymerase chain reaction (PCR)-generated DNA containing up to 585 CAG repeats using atomic force microscopy (AFM). As the number of CAG repeats increased, an increasing proportion of the DNA molecules exhibited unusual structural features, including convolutions and multiple protrusions. At least some of these features are hairpin loops, as judged by cross-sectional analysis and sensitivity to cleavage by mung bean nuclease. Single-molecule force measurements showed that the convoluted DNA was very resistant to untangling. In vitro replication by PCR was markedly reduced, and TseI restriction enzyme digestion was also hindered by the abnormal DNA structures. However, significantly, the DNA gained sensitivity to cleavage by the Type III restriction-modification enzyme, EcoP15I. CONCLUSIONS/SIGNIFICANCE: "Super-long" CAG repeats are found in a number of neurological diseases and may also appear through CAG repeat instability. We suggest that unusual DNA structures associated with super-long CAG repeats decrease transcriptional efficiency in vitro. We also raise the possibility that if these structures occur in vivo, they may play a role in the aetiology of CAG repeat diseases such as HD
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