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    AuthorTobin, Desmond J. (3)Whitaker, David J. (3)Bloj, Marina (2)Denyer, Morgan C.T. (2)Harris, J.M. (2)Heron, James (2)Singh, Suman K. (2)Youseffi, Mansour (2)Aaen-Stockdale, Craig (1)Abbas, Waqas A. (1)View MoreSubject
    Humans (17)
    REF 2014 (16)Adult (9)Female (8)Male (7)Middle aged (4)Aged (3)Auditory perception (2)Cell line (2)Cues (2)View MoreDate Issued2013 (1)2012 (5)2011 (2)2010 (3)2009 (1)2008 (4)2004 (1)

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    Duration channels mediate human time perception

    Heron, James; Aaen-Stockdale, Craig; Hotchkiss, John; Roach, N.W.; McGraw, Paul V.; Whitaker, David J. (2012)
    The task of deciding how long sensory events seem to last is one that the human nervous system appears to perform rapidly and, for sub-second intervals, seemingly without conscious effort. That these estimates can be performed within and between multiple sensory and motor domains suggest time perception forms one of the core, fundamental processes of our perception of the world around us. Given this significance, the current paucity in our understanding of how this process operates is surprising. One candidate mechanism for duration perception posits that duration may be mediated via a system of duration-selective 'channels', which are differentially activated depending on the match between afferent duration information and the channels' 'preferred' duration. However, this model awaits experimental validation. In the current study, we use the technique of sensory adaptation, and we present data that are well described by banks of duration channels that are limited in their bandwidth, sensory-specific, and appear to operate at a relatively early stage of visual and auditory sensory processing. Our results suggest that many of the computational principles the nervous system applies to coding visual spatial and auditory spectral information are common to its processing of temporal extent.
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    Elevated activity and microglial expression of myeloperoxidase in demyelinated cerebral cortex in multiple sclerosis

    Gray, E.; Thomas, T. L.; Betmouni, S.; Scolding, N.; Love, S. (2008)
    Recent studies have revealed extensive cortical demyelination in patients with progressive multiple sclerosis (MS). Demyelination in gray matter lesions is associated with activation of microglia. Macrophages and microglia are known to express myeloperoxidase (MPO) and generate reactive oxygen species during myelin phagocytosis in the white matter. In the present study we examined the extent of microglial activation in the cerebral cortex and the relationship of microglial activation and MPO activity to cortical demyelination. Twenty-one cases of neuropathologically confirmed multiple sclerosis, with 34 cortical lesions, were used to assess microglial activation. HLA-DR immunolabeling of activated microglia was significantly higher in demyelinated MS cortex than control cortex and, within the MS cohort, was significantly greater within cortical lesions than in matched non-demyelinated areas of cortex. In homogenates of MS cortex, cortical demyelination was associated with significantly elevated MPO activity. Immunohistochemistry revealed MPO in CD68-positive microglia within cortical plaques, particularly toward the edge of the plaques, but not in microglia in adjacent non-demyelinated cortex. Cortical demyelination in MS is associated with increased activity of MPO, which is expressed by a CD68-positive subset of activated microglia, suggesting that microglial production of reactive oxygen species is likely to be involved in cortical demyelination.
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    Recalibration of perceived time across sensory modalities

    Hanson, James Vincent Michael; Heron, James; Whitaker, David J. (2008)
    When formulating an estimate of event time, the human sensory system has been shown to possess a degree of perceptual flexibility. Specifically, the perceived relative timing of auditory and visual stimuli is, to some extent, a product of recent experience. It has been suggested that this form of sensory recalibration may be peculiar to the audiovisual domain. Here we investigate how adaptation to sensory asynchrony influences the perceived temporal order of audiovisual, audiotactile and visuotactile stimulus pairs. Our data show that a brief period of repeated exposure to asynchrony in any of these sensory pairings results in marked changes in subsequent temporal order judgments: the point of perceived simultaneity shifts toward the level of adaptation asynchrony. We find that the size and nature of this shift is very similar in all three pairings and that sensitivity to asynchrony is unaffected by the adaptation process. In light of these findings we suggest that a single supramodal mechanism may be responsible for the observed recalibration of multisensory perceived time.
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    Development, validation and application of a patient satisfaction scale for a community pharmacy medicines-management service

    Tinelli, M.; Blenkinsopp, Alison; Bond, C. (2011)
    OBJECTIVE: To develop, validate and apply a scale to measure patient satisfaction in a randomised controlled trial of community pharmacy service. METHODS: Published scales were reviewed to inform development of the patient satisfaction scale. Questionnaires were sent to patients in the control (n=500) and intervention (n=941) groups of a randomised controlled trial of community pharmacy-led management of coronary heart disease at baseline and 12-month follow-up. Any underlying main factors were assessed with exploratory factor analysis. Reliability and construct validity were tested. The 15-item scale was used to compare patient satisfaction across arms with their most recent pharmacy visit. RESULTS: Response rates were 92% (461/500) for control and 96% (903/941) for intervention groups at baseline and 85% control (399/472) and intervention (810/941) at follow-up. At baseline satisfaction was very similar in the intervention and control groups (median scores of 42). At follow-up mean satisfaction had significantly improved for the intervention compared with the control (median scores of 46 compared with 43; P<0.01); intervention females were more likely to be satisfied with the service than males (49 compared with 44; P<0.01). Three main factors explained the majority of the data variance. Cronbach's alpha was 0.7-0.9 for both groups over time for all factors and total scale. An increase in the overall satisfaction corresponding to a decrease in subjects wanting that particular service to be provided during their next visit indicated construct validity of the scale. CONCLUSION: A new scale of patient satisfaction with community pharmacy services was developed and shown to be reliable and valid. Its application showed increased satisfaction in the intervention group receiving a new pharmacy service.
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    Imaging of the cell surface interface using objective coupled widefield surface plasmon microscopy

    Jamil, M. Mahadi Abdul; Denyer, Morgan C.T.; Youseffi, Mansour; Britland, Stephen T.; Liu, S.; See, C.W.; Somekh, M.G.; Zhang, J. (2008)
    We report on the development and on the first use of the widefield surface plasmon (WSPR) microscope in the examination of the cell surface interface at submicron lateral resolutions. The microscope is Kohler illuminated and uses either a 1.45 numerical aperture (NA) oil immersion lens, or a 1.65 NA oil immersion lens to excite surface plasmons at the interface between a thin gold layer and a glass or sapphire cover slip. Like all surface plasmon microscope systems the WSPR has been proven in previous studies to also be capable of nanometric z-scale resolutions. In this study we used the system to image the interface between HaCaT cells and the gold layer. Imaging was performed in air using fixed samples and the 1.45 NA objective based system and also using live cells in culture media using the 1.65 NA based system. Imaging in air enabled the visualisation of high resolution and high-contrast submicron features identified by vinculin immunostaining as component of focal contacts and focal adhesions. In comparison, imaging in fluid enabled cell surface interfacial interactions to be tracked by time-lapse video WSPR microscopy. Our results indicate that the cell surface interface and thus cell signalling mechanisms may be readily interrogated in live cells without the use of labelling techniques.
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    An interaction between KSHV ORF57 and UIF provides mRNA-adaptor redundancy in herpesvirus intronless mRNA export

    Jackson, B.R.; Boyne, James R.; Noerenberg, M.; Taylor, A.; Hautbergue, G.M.; Walsh, M.J.; Wheat, R.; Blackbourn, D.J.; Wilson, S.A.; Whitehouse, A. (2011)
    The hTREX complex mediates cellular bulk mRNA nuclear export by recruiting the nuclear export factor, TAP, via a direct interaction with the export adaptor, Aly. Intriguingly however, depletion of Aly only leads to a modest reduction in cellular mRNA nuclear export, suggesting the existence of additional mRNA nuclear export adaptor proteins. In order to efficiently export Kaposi's sarcoma-associated herpesvirus (KSHV) intronless mRNAs from the nucleus, the KSHV ORF57 protein recruits hTREX onto viral intronless mRNAs allowing access to the TAP-mediated export pathway. Similarly however, depletion of Aly only leads to a modest reduction in the nuclear export of KSHV intronless mRNAs. Herein, we identify a novel interaction between ORF57 and the cellular protein, UIF. We provide the first evidence that the ORF57-UIF interaction enables the recruitment of hTREX and TAP to KSHV intronless mRNAs in Aly-depleted cells. Strikingly, depletion of both Aly and UIF inhibits the formation of an ORF57-mediated nuclear export competent ribonucleoprotein particle and consequently prevents ORF57-mediated mRNA nuclear export and KSHV protein production. Importantly, these findings highlight that redundancy exists in the eukaryotic system for certain hTREX components involved in the mRNA nuclear export of intronless KSHV mRNAs.
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    Bone morphogenetic proteins differentially regulate pigmentation in human skin cells

    Singh, Suman K.; Abbas, Waqas A.; Tobin, Desmond J. (2012)
    Bone morphogenetic proteins (BMPs) are a large family of multi-functional secreted signalling molecules. Previously BMP2/4 were shown to inhibit skin pigmentation by downregulating tyrosinase expression and activity in epidermal melanocytes. However, a possible role for other BMP family members and their antagonists in melanogenesis has not yet been explored. In this study we show that BMP4 and BMP6, from two different BMP subclasses, and their antagonists noggin and sclerostin were variably expressed in melanocytes and keratinocytes in human skin. We further examined their involvement in melanogenesis and melanin transfer using fully matched primary cultures of adult human melanocytes and keratinocytes. BMP6 markedly stimulated melanogenesis by upregulating tyrosinase expression and activity, and also stimulated the formation of filopodia and Myosin-X expression in melanocytes, which was associated with increased melanosome transfer from melanocytes to keratinocytes. BMP4, by contrast, inhibited melanin synthesis and transfer to below baseline levels. These findings were confirmed using siRNA knockdown of BMP receptors BMPR1A/1B or of Myosin-X, as well as by incubating cells with the antagonists noggin and sclerostin. While BMP6 was found to use the p38MAPK pathway to regulate melanogenesis in human melanocytes independently of the Smad pathway, p38MAPK, PI3-K and Smad pathways were all involved in BMP6-mediated melanin transfer. This suggests that pigment formation may be regulated independently of pigment transfer. These data reveal a complex involvement of regulation of different members of the BMP family, their antagonists and inhibitory Smads, in melanocytes behaviour.
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    Development of a novel liquid crystal based cell traction force transducer system

    Soon, Chin Fhong; Youseffi, Mansour; Berends, Rebecca F.; Blagden, Nicholas; Denyer, Morgan C.T. (2013)
    Keratinocyte traction forces play a crucial role in wound healing. The aim of this study was to develop a novel cell traction force (CTF) transducer system based on cholesteryl ester liquid crystals (LC). Keratinocytes cultured on LC induced linear and isolated deformation lines in the LC surface. As suggested by the fluorescence staining, the deformation lines appeared to correlate with the forces generated by the contraction of circumferential actin filaments which were transmitted to the LC surface via the focal adhesions. Due to the linear viscoelastic behavior of the LC, Hooke's equation was used to quantify the CTFs by associating Young's modulus of LC to the cell induced stresses and biaxial strain in forming the LC deformation. Young's modulus of the LC was profiled by using spherical indentation and determined at approximately 87.1+/-17.2kPa. A new technique involving cytochalasin-B treatment was used to disrupt the intracellular force generating actin fibers, and consequently the biaxial strain in the LC induced by the cells was determined. Due to the improved sensitivity and spatial resolution ( approximately 1mum) of the LC based CTF transducer, a wide range of CTFs was determined (10-120nN). These were found to be linearly proportional to the length of the deformations. The linear relationship of CTF-deformations was then applied in a bespoke CTF mapping software to estimate CTFs and to map CTF fields. The generated CTF map highlighted distinct distributions and different magnitude of CTFs were revealed for polarized and non-polarized keratinocytes.
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    The acoustic and visual factors influencing the construction of tranquil space in urban and rural environments tranquil spaces-quiet places?

    Pheasant, Robert J.; Horoshenkov, Kirill V.; Watts, Gregory R.; Barrett, Brendan T. (2008)
    Prior to this work no structured mechanism existed in the UK to evaluate the tranquillity of open spaces with respect to the characteristics of both acoustic and visual stimuli. This is largely due to the fact that within the context of "tranquil" environments, little is known about the interaction of the audio-visual modalities and how they combine to lead to the perception of tranquillity. This paper presents the findings of a study in which visual and acoustic data, captured from 11 English rural and urban landscapes, were used by 44 volunteers to make subjective assessments of both their perceived tranquillity of a location, and the loudness of five generic soundscape components. The results were then analyzed alongside objective measurements taken in the laboratory. It was found that the maximum sound pressure level (L(Amax)) and the percentage of natural features present at a location were the key factors influencing tranquillity. Engineering formulas for the tranquillity as a function of the noise level and proportion of the natural features are proposed.
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    Enhanced DNA binding capacity on up-regulated epidermal wild-type p53 in vitiligo by H2O2-mediated oxidation: a possible repair mechanism for DNA damage

    Salem, Mohamed M.A.; Shalbaf, Mohammad; Gibbons, Nick C.; Chavan, Bhavan; Thornton, M. Julie; Schallreuter, Karin U. (2009)
    Vitiligo is characterized by a patchy loss of inherited skin color affecting approximately 0.5% of individuals of all races. Despite the absence of the protecting pigment and the overwhelming evidence for hydrogen peroxide (H(2)O(2))-induced oxidative stress in the entire epidermis of these patients, there is neither increased photodamage/skin aging nor a higher incidence for sun-induced nonmelanoma skin cancer. Here we demonstrate for the first time increased DNA damage via 8-oxoguanine in the skin and plasma in association with epidermal up-regulated phosphorylated/acetylated p53 and high levels of the p53 antagonist p76(MDM2). Short-patch base-excision repair via hOgg1, APE1, and polymerasebeta DNA repair is up-regulated. Overexpression of Bcl-2 and low caspase 3 and cytochrome c levels argue against increased apoptosis in this disease. Moreover, we show the presence of high epidermal peroxynitrite (ONOO(-)) levels via nitrotyrosine together with high nitrated p53 levels. We demonstrate by EMSA that nitration of p53 by ONOO(-) (300 x 10(-6) M) abrogates DNA binding, while H(2)O(2)-oxidized p53 (10(-3) M) enhances DNA binding capacity and prevents ONOO(-)-induced abrogation of DNA binding. Taken together, we add a novel reactive oxygen species to the list of oxidative stress inducers in vitiligo. Moreover, we propose up-regulated wild-type p53 together with p76(MDM2) as major players in the control of DNA damage/repair and prevention of photodamage and nonmelanoma skin cancer in vitiligo.
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