949 resultados para Tight junction


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We synthesized nanoscale TiO2-RuO2 alloys by atomic layer deposition (ALD) that possess a high work function and are highly conductive. As such, they function as good Schottky contacts to extract photogenerated holes from n-type silicon while simultaneously interfacing with water oxidation catalysts. The ratio of TiO2 to RuO2 can be precisely controlled by the number of ALD cycles for each precursor. Increasing the composition above 16% Ru sets the electronic conductivity and the metal work function. No significant Ohmic loss for hole transport is measured as film thickness increases from 3 to 45 nm for alloy compositions >= 16% Ru. Silicon photoanodes with a 2 nm SiO2 layer that are coated by these alloy Schottky contacts having compositions in the range of 13-46% Ru exhibit average photovoltages of 525 mV, with a maximum photovoltage of 570 mV achieved. Depositing TiO2-RuO2 alloys on nSi sets a high effective work function for the Schottky junction with the semiconductor substrate, thus generating a large photovoltage that is isolated from the properties of an overlying oxygen evolution catalyst or protection layer.

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Hepatitis C virus is a positive-sense single-stranded RNA virus. The gene junction partitioning the viral glycoproteins E1 and E2 displays concurrent sequence evolution with the 3′-end of E1 highly conserved and the 5′-end of E2 highly heterogeneous. This gene junction is also believed to contain structured RNA elements, with a growing body of evidence suggesting that such structures can act as an additional level of viral replication and transcriptional control. We have previously used ultradeep pyrosequencing to analyze an amplicon library spanning the E1/E2 gene junction from a treatment naïve patient where samples were collected over 10 years of chronic HCV infection. During this timeframe maintenance of an in-frame insertion, recombination and humoral immune targeting of discrete virus sub-populations was reported. In the current study, we present evidence of epistatic evolution across the E1/E2 gene junction and observe the development of co-varying networks of codons set against a background of a complex virome with periodic shifts in population dominance. Overtime, the number of codons actively mutating decreases for all virus groupings. We identify strong synonymous co-variation between codon sites in a group of sequences harbouring a 3 bp in-frame insertion and propose that synonymous mutation acts to stabilize the RNA structural backbone.

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With increasingly complex engineering assets and tight economic requirements, asset reliability becomes more crucial in Engineering Asset Management (EAM). Improving the reliability of systems has always been a major aim of EAM. Reliability assessment using degradation data has become a significant approach to evaluate the reliability and safety of critical systems. Degradation data often provide more information than failure time data for assessing reliability and predicting the remnant life of systems. In general, degradation is the reduction in performance, reliability, and life span of assets. Many failure mechanisms can be traced to an underlying degradation process. Degradation phenomenon is a kind of stochastic process; therefore, it could be modelled in several approaches. Degradation modelling techniques have generated a great amount of research in reliability field. While degradation models play a significant role in reliability analysis, there are few review papers on that. This paper presents a review of the existing literature on commonly used degradation models in reliability analysis. The current research and developments in degradation models are reviewed and summarised in this paper. This study synthesises these models and classifies them in certain groups. Additionally, it attempts to identify the merits, limitations, and applications of each model. It provides potential applications of these degradation models in asset health and reliability prediction.

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Sex hormone-binding globulin (SHBG) is a homodimeric plasma glycoprotein that is the major sex steroid carrier-protein in the bloodstream and functions also as a key regulator of steroid bioavailability within target tissues, such as the prostate. Additionally, SHBG binds to prostatic cell membranes via the putative and unidentified SHBG receptor (RSHBG), activating a signal transduction pathway implicated in stimulating both proliferation and expression of prostate specific antigen (PSA) in prostate cell lines in vitro. A yeast-two hybrid assay suggested an interaction between SHBG and kallikrein-related protease (KLK) 4, which is a serine protease implicated in the progression of prostate cancer. The potential interaction between these two proteins was investigated in this PhD thesis to determine whether SHBG is a proteolytic substrate of KLK4 and other members of the KLK family including KLK3/PSA, KLK7 and KLK14. Furthermore, the effects from SHBG proteolytic degradation on SHBG-regulated steroid bioavailability and the activation of the putative RSHBG signal transduction pathway were examined in the LNCaP prostate cancer cell line. SHBG was found to be a proteolytic substrate of the trypsin-like KLK4 and KLK14 in vitro, yielding several proteolysis fragments. Both chymotrypsin-like PSA and KLK7 displayed insignificant proteolytic activity against SHBG. The kinetic parameters of SHBG proteolysis by KLK4 and KLK14 demonstrate a strong enzyme-substrate binding capacity, possessing a Km of 1.2 ± 0.7 µM and 2.1 ± 0.6 µM respectively. The catalytic efficiencies (kcat/Km) of KLK4 and KLK14 proteolysis of SHBG were 1.6 x 104 M-1s-1 and 3.8 x 104 M-1s-1 respectively, which were comparable to parameters previously reported for peptide substrates. N-terminal sequencing of the fragments revealed cleavage near the junction of the N- and C-terminal laminin globulin-like (G-like) domains of SHBG, resulting in the division of the two globulins and ultimately the full degradation of these fragments by KLK4 and KLK14 over time. Proteolytic fragments that may retain steroid binding were rapidly degraded by both proteases, while fragments containing residues beyond the steroid binding pocket were less degraded over the same period of time. Degradation of SHBG was inhibited by the divalent metal cations calcium and zinc for KLK4, and calcium, zinc and magnesium for KLK14. The human secreted serine protease inhibitors (serpins), α1-antitrypsin and α2-antiplasmin, inhibited KLK4 and KLK14 proteolysis of SHBG; α1-antichymotrypsin inhibited KLK4 but not KLK14 activity. The inhibition by these serpins was comparable and in some cases more effective than general trypsin protease inhibitors such as aprotinin and phenylmethanesulfonyl fluoride (PMSF). The binding of 5α-dihydrotestosterone (DHT) to SHBG modulated interactions with KLK4 and KLK14. Steroid-free SHBG was more readily digested by both enzymes than DHT-bound SHBG. Moreover, a binding interaction exists between SHBG and pro-KLK4 and pro-KLK14, with DHT strengthening the binding to pro-KLK4 only. The inhibition of androgen uptake by cultured prostate cancer cells, mediated by SHBG steroid-binding, was examined to assess whether SHBG proteolysis by KLK4 and KLK14 modulated this process. Proteolytic digestion eliminated the ability of SHBG to inhibit the uptake of DHT from conditioned media into LNCaP cells. Therefore, the proteolysis of SHBG by KLK4 and KLK14 increased steroid bioavailability in vitro, leading to an increased uptake of androgens by prostate cancer cells. Interestingly, different transcriptional responses of PSA and KLK2, which are androgen-regulated genes, to DHT-bounsd SHBG treatment were observed between low and high passage number LNCaP cells (lpLNCaP and hpLNCaP respectively). HpLNCaP cells treated with DHT-bound SHBG demonstrated a significant synergistic upregulation of PSA and KLK2 above DHT or SHBG treatment alone, which is similar to previously reported downstream responses from RSHBG-mediated signaling activation. As this result was not seen in lpLNCaP cells, only hpLNCaP cells were further investigated to examine the modulation of potential RSHBG activity by KLK4 and KLK14 proteolysis of SHBG. Contrary to reported results, no increase in intracellular cAMP was observed in hpLNCaP cells when treated with SHBG in the presence and absence of either DHT or estradiol. As a result, the modulation of RSHBG-mediated signaling activation could not be determined. Finally, the identification of the RSHBG from both breast (MCF-7) and prostate cancer (LNCaP) cell lines was attempted. Fluorescently labeled peptides corresponding to the putative receptor binding domain (RBD) of SHBG were shown to be internalized by MCF-7 cells. Crosslinking of the RBD peptide to the cell surfaces of both MCF-7 and LNCaP cells, demonstrated the interaction of the peptide with several targets. These targets were then captured using RBD peptides synthesized onto a hydrophilic scaffold and analysed by mass spectrometry. The samples captured by the RBD peptide returned statistically significantly matches for cytokeratin 8, 18 and 19 as well as microtubule-actin crosslinking factor 1, which may indicate a novel interaction between SHBG and these proteins, but ultimately failed to detect a membrane receptor potentially responsible for the putative RSHBG-mediated signaling. This PhD project has reported the proteolytic processing of SHBG by two members of the kallikrein family, KLK4 and KLK14. The effect of SHBG proteolysis by KLK4 and KLK14 on RSHBG-mediated signaling activation was unable to be determined as the reported signal transduction pathway was not activated after treatment with SHBG, in combination with either DHT or estradiol. However, the digestion of SHBG by these two proteases positively regulated androgen bioavailability to prostate cancer cells in vitro. The increased uptake of androgens is deleterious in prostate cancer due to the promotion of proliferation, metastasis, invasion and the inhibition of apoptosis. The increased bioavailability of androgens, from SHBG proteolysis by KLK4 and KLK14, may therefore promote both carcinogenesis and progression of prostate cancer. Finally, this information may contribute to the development of therapeutic treatment strategies for prostate cancer by inhibiting the proteolysis of SHBG, by KLK4 and KLK14, to prevent the increased uptake of androgens by hormone-dependent cancerous tissues.

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The degradation of high voltage electrical insulation is a prime factor that can significantly influence the reliability performance and the costs of maintaining high voltage electricity networks. Little information is known about the system of localized degradation from corona discharges on the relatively new silicone rubber sheathed composite insulators that are now being widely used in high voltage applications. This current work focuses on the fundamental principles of electrical corona discharge phenomena to provide further insights to where damaging surface discharges may localize and examines how these discharges may degrade the silicone rubber material. Although water drop corona has been identified by many authors as a major cause of deterioration of silicone rubber high voltage insulation until now no thorough studies have been made of this phenomenon. Results from systematic measurements taken using modern digital instrumentation to simultaneously record the discharge current pulses and visible images associated with corona discharges from between metal electrodes, metal electrodes and water drops, and between waters drops on the surface of silicone rubber insulation, using a range of 50 Hz voltages are inter compared. Visual images of wet electrodes show how water drops can play a part in encouraging flashover, and the first reproducible visual images of water drop corona at the triple junction of water air and silicone rubber insulation are presented. A study of the atomic emission spectra of the corona produced by the discharge from its onset up to and including spark-over, using a high resolution digital spectrometer with a fiber optic probe, provides further understanding of the roles of the active species of atoms and molecules produced by the discharge that may be responsible for not only for chemical changes of insulator surfaces, but may also contribute to the degradation of the metal fittings that support the high voltage insulators. Examples of real insulators and further work specific to the electrical power industry are discussed. A new design concept to prevent/reduce the damaging effects of water drop corona is also presented.

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Over the past decade, plants have been used as expression hosts for the production of pharmaceutically important and commercially valuable proteins. Plants offer many advantages over other expression systems such as lower production costs, rapid scale up of production, similar post-translational modification as animals and the low likelihood of contamination with animal pathogens, microbial toxins or oncogenic sequences. However, improving recombinant protein yield remains one of the greatest challenges to molecular farming. In-Plant Activation (InPAct) is a newly developed technology that offers activatable and high-level expression of heterologous proteins in plants. InPAct vectors contain the geminivirus cis elements essential for rolling circle replication (RCR) and are arranged such that the gene of interest is only expressed in the presence of the cognate viral replication-associated protein (Rep). The expression of Rep in planta may be controlled by a tissue-specific, developmentally regulated or chemically inducible promoter such that heterologous protein accumulation can be spatially and temporally controlled. One of the challenges for the successful exploitation of InPAct technology is the control of Rep expression as even very low levels of this protein can reduce transformation efficiency, cause abnormal phenotypes and premature activation of the InPAct vector in regenerated plants. Tight regulation over transgene expression is also essential if expressing cytotoxic products. Unfortunately, many tissue-specific and inducible promoters are unsuitable for controlling expression of Rep due to low basal activity in the absence of inducer or in tissues other than the target tissue. This PhD aimed to control Rep activity through the production of single chain variable fragments (scFvs) specific to the motif III of Tobacco yellow dwarf virus (TbYDV) Rep. Due to the important role played by the conserved motif III in the RCR, it was postulated that such scFvs can be used to neutralise the activity of the low amount of Rep expressed from a “leaky” inducible promoter, thus preventing activation of the TbYDV-based InPAct vector until intentional induction. Such scFvs could also offer the potential to confer partial or complete resistance to TbYDV, and possibly heterologous viruses as motif III is conserved between geminiviruses. Studies were first undertaken to determine the levels of TbYDV Rep and TbYDV replication-associated protein A (RepA) required for optimal transgene expression from a TbYDV-based InPAct vector. Transient assays in a non-regenerable Nicotiana tabacum (NT-1) cell line were undertaken using a TbYDV-based InPAct vector containing the uidA reporter gene (encoding GUS) in combination with TbYDV Rep and RepA under the control of promoters with high (CaMV 35S) or low (Banana bunchy top virus DNA-R, BT1) activity. The replication enhancer protein of Tomato leaf curl begomovirus (ToLCV), REn, was also used in some co-bombardment experiments to examine whether RepA could be substituted by a replication enhancer from another geminivirus genus. GUS expression was observed both quantitatively and qualitatively by fluorometric and histochemical assays, respectively. GUS expression from the TbYDV-based InPAct vector was found to be greater when Rep was expected to be expressed at low levels (BT1 promoter) rather than high levels (35S promoter). GUS expression was further enhanced when Rep and RepA were co-bombarded with a low ratio of Rep to RepA. Substituting TbYDV RepA with ToLCV REn also enhanced GUS expression but more importantly highest GUS expression was observed when cells were co-transformed with expression vectors directing low levels of Rep and high levels of RepA irrespective of the level of REn. In this case, GUS expression was approximately 74-fold higher than that from a non-replicating vector. The use of different terminators, namely CaMV 35S and Nos terminators, in InPAct vectors was found to influence GUS expression. In the presence of Rep, GUS expression was greater using pInPActGUS-Nos rather than pInPActGUS-35S. The only instance of GUS expression being greater from vectors containing the 35S terminator was when comparing expression from cells transformed with Rep, RepA and REnexpressing vectors and either non-replicating vectors, p35SGS-Nos or p35SGS-35S. This difference was most likely caused by an interaction of viral replication proteins with each other and the terminators. These results indicated that (i) the level of replication associated proteins is critical to high transgene expression, (ii) the choice of terminator within the InPAct vector may affect expression levels and (iii) very low levels of Rep can activate InPAct vectors hence controlling its activity is critical. Prior to generating recombinant scFvs, a recombinant TbYDV Rep was produced in E. coli to act as a control to enable the screening for Rep-specific antibodies. A bacterial expression vector was constructed to express recombinant TbYDV Rep with an Nterminal His-tag (N-His-Rep). Despite investigating several purification techniques including Ni-NTA, anion exchange, hydrophobic interaction and size exclusion chromatography, N-His-Rep could only be partially purified using a Ni-NTA column under native conditions. Although it was not certain that this recombinant N-His-Rep had the same conformation as the native TbYDV Rep and was functional, results from an electromobility shift assay (EMSA) showed that N-His-Rep was able to interact with the TbYDV LIR and was, therefore, possibly functional. Two hybridoma cell lines from mice, immunised with a synthetic peptide containing the TbYDV Rep motif III amino acid sequence, were generated by GenScript (USA). Monoclonal antibodies secreted by the two hybridoma cell lines were first screened against denatured N-His-Rep in Western analysis. After demonstrating their ability to bind N-His-Rep, two scFvs (scFv1 and scFv2) were generated using a PCR-based approach. Whereas the variable heavy chain (VH) from both cell lines could be amplified, only the variable light chain (VL) from cell line 2 was amplified. As a result, scFv1 contained VH and VL from cell line 1, whereas scFv2 contained VH from cell line 2 and VL from cell line 1. Both scFvs were first expressed in E. coli in order to evaluate their affinity to the recombinant TbYDV N-His-Rep. The preliminary results demonstrated that both scFvs were able to bind to the denatured N-His-Rep. However, EMSAs revealed that only scFv2 was able to bind to native N-His-Rep and prevent it from interacting with the TbYDV LIR. Each scFv was cloned into plant expression vectors and co-bombarded into NT-1 cells with the TbYDV-based InPAct GUS expression vector and pBT1-Rep to examine whether the scFvs could prevent Rep from mediating RCR. Although it was expected that the addition of the scFvs would result in decreased GUS expression, GUS expression was found to slightly increase. This increase was even more pronounced when the scFvs were targeted to the cell nucleus by the inclusion of the Simian virus 40 large T antigen (SV40) nuclear localisation signal (NLS). It was postulated that the scFvs were binding to a proportion of Rep, leaving a small amount available to mediate RCR. The outcomes of this project provide evidence that very high levels of recombinant protein can theoretically be expressed using InPAct vectors with judicious selection and control of viral replication proteins. However, the question of whether the scFvs generated in this project have sufficient affinity for TbYDV Rep to prevent its activity in a stably transformed plant remains unknown. It may be that other scFvs with different combinations of VH and VL may have greater affinity for TbYDV Rep. Such scFvs, when expressed at high levels in planta, might also confer resistance to TbYDV and possibly heterologous geminiviruses.

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The inquiry documented in this thesis is located at the nexus of technological innovation and traditional schooling. As we enter the second decade of a new century, few would argue against the increasingly urgent need to integrate digital literacies with traditional academic knowledge. Yet, despite substantial investments from governments and businesses, the adoption and diffusion of contemporary digital tools in formal schooling remain sluggish. To date, research on technology adoption in schools tends to take a deficit perspective of schools and teachers, with the lack of resources and teacher ‘technophobia’ most commonly cited as barriers to digital uptake. Corresponding interventions that focus on increasing funding and upskilling teachers, however, have made little difference to adoption trends in the last decade. Empirical evidence that explicates the cultural and pedagogical complexities of innovation diffusion within long-established conventions of mainstream schooling, particularly from the standpoint of students, is wanting. To address this knowledge gap, this thesis inquires into how students evaluate and account for the constraints and affordances of contemporary digital tools when they engage with them as part of their conventional schooling. It documents the attempted integration of a student-led Web 2.0 learning initiative, known as the Student Media Centre (SMC), into the schooling practices of a long-established, high-performing independent senior boys’ school in urban Australia. The study employed an ‘explanatory’ two-phase research design (Creswell, 2003) that combined complementary quantitative and qualitative methods to achieve both breadth of measurement and richness of characterisation. In the initial quantitative phase, a self-reported questionnaire was administered to the senior school student population to determine adoption trends and predictors of SMC usage (N=481). Measurement constructs included individual learning dispositions (learning and performance goals, cognitive playfulness and personal innovativeness), as well as social and technological variables (peer support, perceived usefulness and ease of use). Incremental predictive models of SMC usage were conducted using Classification and Regression Tree (CART) modelling: (i) individual-level predictors, (ii) individual and social predictors, and (iii) individual, social and technological predictors. Peer support emerged as the best predictor of SMC usage. Other salient predictors include perceived ease of use and usefulness, cognitive playfulness and learning goals. On the whole, an overwhelming proportion of students reported low usage levels, low perceived usefulness and a lack of peer support for engaging with the digital learning initiative. The small minority of frequent users reported having high levels of peer support and robust learning goal orientations, rather than being predominantly driven by performance goals. These findings indicate that tensions around social validation, digital learning and academic performance pressures influence students’ engagement with the Web 2.0 learning initiative. The qualitative phase that followed provided insights into these tensions by shifting the analytics from individual attitudes and behaviours to shared social and cultural reasoning practices that explain students’ engagement with the innovation. Six indepth focus groups, comprising 60 students with different levels of SMC usage, were conducted, audio-recorded and transcribed. Textual data were analysed using Membership Categorisation Analysis. Students’ accounts converged around a key proposition. The Web 2.0 learning initiative was useful-in-principle but useless-in-practice. While students endorsed the usefulness of the SMC for enhancing multimodal engagement, extending peer-topeer networks and acquiring real-world skills, they also called attention to a number of constraints that obfuscated the realisation of these design affordances in practice. These constraints were cast in terms of three binary formulations of social and cultural imperatives at play within the school: (i) ‘cool/uncool’, (ii) ‘dominant staff/compliant student’, and (iii) ‘digital learning/academic performance’. The first formulation foregrounds the social stigma of the SMC among peers and its resultant lack of positive network benefits. The second relates to students’ perception of the school culture as authoritarian and punitive with adverse effects on the very student agency required to drive the innovation. The third points to academic performance pressures in a crowded curriculum with tight timelines. Taken together, findings from both phases of the study provide the following key insights. First, students endorsed the learning affordances of contemporary digital tools such as the SMC for enhancing their current schooling practices. For the majority of students, however, these learning affordances were overshadowed by the performative demands of schooling, both social and academic. The student participants saw engagement with the SMC in-school as distinct from, even oppositional to, the conventional social and academic performance indicators of schooling, namely (i) being ‘cool’ (or at least ‘not uncool’), (ii) sufficiently ‘compliant’, and (iii) achieving good academic grades. Their reasoned response therefore, was simply to resist engagement with the digital learning innovation. Second, a small minority of students seemed dispositionally inclined to negotiate the learning affordances and performance constraints of digital learning and traditional schooling more effectively than others. These students were able to engage more frequently and meaningfully with the SMC in school. Their ability to adapt and traverse seemingly incommensurate social and institutional identities and norms is theorised as cultural agility – a dispositional construct that comprises personal innovativeness, cognitive playfulness and learning goals orientation. The logic then is ‘both and’ rather than ‘either or’ for these individuals with a capacity to accommodate both learning and performance in school, whether in terms of digital engagement and academic excellence, or successful brokerage across multiple social identities and institutional affiliations within the school. In sum, this study takes us beyond the familiar terrain of deficit discourses that tend to blame institutional conservatism, lack of resourcing and teacher resistance for low uptake of digital technologies in schools. It does so by providing an empirical base for the development of a ‘third way’ of theorising technological and pedagogical innovation in schools, one which is more informed by students as critical stakeholders and thus more relevant to the lived culture within the school, and its complex relationship to students’ lives outside of school. It is in this relationship that we find an explanation for how these individuals can, at the one time, be digital kids and analogue students.

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Background: It has been proposed that adenosine triphosphate (ATP) released from red blood cells (RBCs) may contribute to the tight coupling between blood flow and oxygen demand in contracting skeletal muscle. To determine whether ATP may contribute to the vasodilatory response to exercise in the forearm, we measured arterialised and venous plasma ATP concentration and venous oxygen content in 10 healthy young males at rest, and at 30 and 180 seconds during dynamic handgrip exercise at 45% of maximum voluntary contraction (MVC). Results: Venous plasma ATP concentration was elevated above rest after 30 seconds of exercise (P < 0.05), and remained at this higher level 180 seconds into exercise (P < 0.05 versus rest). The increase in ATP was mirrored by a decrease in venous oxygen content. While there was no significant relationship between ATP concentration and venous oxygen content at 30 seconds of exercise, they were moderately and inversely correlated at 180 seconds of exercise (r = -0.651, P = 0.021). Arterial ATP concentration remained unchanged throughout exercise, resulting in an increase in the venous-arterial ATP difference. Conclusions: Collectively these results indicate that ATP in the plasma originated from the muscle microcirculation, and are consistent with the notion that deoxygenation of the blood perfusing the muscle acts as a stimulus for ATP release. That ATP concentration was elevated just 30 seconds after the onset of exercise also suggests that ATP may be a contributing factor to the blood flow response in the transition from rest to steady state exercise.

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Actuators with deliberately added compliant elements in the transmission system are often described as improving the safety of the actuator at the detriment of the performance. We show that our variant of the Series Elastic Actuator topology, the Velocity Sourced Series Elastic Actuator, has well defined performance characteristics that make for improvements in safety and performance over conventional high impedance actuators. The improvement in performance was principally achieved by having tight velocity control of the DC motor that acts as the mechanical power source for the actuator. Results for performance are given for point to point transition times, while results for safety are based on empirical assessment of the Head Injury Criterion during collisions.

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Current-voltage (I-V) curves of Poly(3-hexyl-thiophene) (P3HT) diodes have been collected to investigate the polymer hole-dominated charge transport. At room temperature and at low electric fields the I-V characteristic is purely Ohmic whereas at medium-high electric fields, experimental data shows that the hole transport is Trap Dominated - Space Charge Limited Current (TD-SCLC). In this regime, it is possible to extract the I-V characteristic of the P3HT/Al junction showing the ideal Schottky diode behaviour over five orders of magnitude. At high-applied electric fields, holes’ transport is found to be in the trap free SCLC regime. We have measured and modelled in this regime the holes’ mobility to evaluate its dependence from the electric field applied and the temperature of the device.

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A point interpolation method with locally smoothed strain field (PIM-LS2) is developed for mechanics problems using a triangular background mesh. In the PIM-LS2, the strain within each sub-cell of a nodal domain is assumed to be the average strain over the adjacent sub-cells of the neighboring element sharing the same field node. We prove theoretically that the energy norm of the smoothed strain field in PIM-LS2 is equivalent to that of the compatible strain field, and then prove that the solution of the PIM- LS2 converges to the exact solution of the original strong form. Furthermore, the softening effects of PIM-LS2 to system and the effects of the number of sub-cells that participated in the smoothing operation on the convergence of PIM-LS2 are investigated. Intensive numerical studies verify the convergence, softening effects and bound properties of the PIM-LS2, and show that the very ‘‘tight’’ lower and upper bound solutions can be obtained using PIM-LS2.

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LiteSteel beam (LSB) is a new cold-formed steel hollow flange channel beam. The unique LSB section is produced by a patented manufacturing process involving simultaneous cold-forming and dual electric resistance welding. To date, limited research has been undertaken on the shear buckling behaviour of LSBs with torsionally rigid, rectangular hollow flanges. For the shear design of LSB web panels, their elastic shear buckling strength must be determined accurately including the potential post-buckling strength. Currently the elastic shear buckling coefficients of web panels are determined by assuming conservatively that the web panels are simply supported at the junction between the flange and web elements. Therefore finite element analyses were carried out to investigate the elastic shear buckling behaviour of LSB sections including the effect of true support conditions at the junction between their flange and web elements. An improved equation for the higher elastic shear buckling coefficient of LSBs was developed and included in the shear capacity equations of Australian cold-formed steel codes. Predicted ultimate shear capacity results were compared with available experimental results, both of which showed considerable improvement to the shear capacities of LSBs. A study on the shear flow distribution of LSBs was also undertaken prior to the elastic buckling analysis study. This paper presents the details of this investigation and the results including the shear flow distribution of LSBs. Keywords: LiteSteel beam, Elastic shear buckling, Shear flow, Cold-formed steel structures, Slender web, Hollow flanges.

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OneSteel Australian Tube Mills has recently developed a new hollow flange channel cold-formed section, known as the LiteSteel Beam (LSB). The innovative LSB sections have the beneficial characteristics of torsionally rigid closed rectangular flanges combined with economical fabrication processes from a single strip of high strength steel. They combine the stability of hot-rolled steel sections with the high strength to weight ratio of conventional cold-formed steel sections. The LSB sections are commonly used as flexural members in residential, industrial and commercial buildings. In order to ensure safe and efficient designs of LSBs, many research studies have been undertaken on the flexural behaviour of LSBs. However, no research has been undertaken on the shear behaviour of LSBs. Therefore this thesis investigated the ultimate shear strength behaviour of LSBs with and without web openings including their elastic buckling and post-buckling characteristics using both experimental and finite element analyses, and developed accurate shear design rules. Currently the elastic shear buckling coefficients of web panels are determined by assuming conservatively that the web panels are simply supported at the junction between the web and flange elements. Therefore finite element analyses were conducted first to investigate the elastic shear buckling behaviour of LSBs to determine the true support condition at the junction between their web and flange elements. An equation for the higher elastic shear buckling coefficient of LSBs was developed and included in the shear capacity equations in the cold-formed steel structures code, AS/NZS 4600. Predicted shear capacities from the modified equations and the available experimental results demonstrated the improvements to the shear capacities of LSBs due to the presence of higher level of fixity at the LSB flange to web juncture. A detailed study into the shear flow distribution of LSB was also undertaken prior to the elastic buckling analysis study. The experimental study of ten LSB sections included 42 shear tests of LSBs with aspect ratios of 1.0 and 1.5 that were loaded at midspan until failure. Both single and back to back LSB arrangements were used. Test specimens were chosen such that all three types of shear failure (shear yielding, inelastic and elastic shear buckling) occurred in the tests. Experimental results showed that the current cold-formed steel design rules are very conservative for the shear design of LSBs. Significant improvements to web shear buckling occurred due to the presence of rectangular hollow flanges while considerable post-buckling strength was also observed. Experimental results were presented and compared with corresponding predictions from the current design rules. Appropriate improvements have been proposed for the shear strength of LSBs based on AISI (2007) design equations and test results. Suitable design rules were also developed under the direct strength method (DSM) format. This thesis also includes the shear test results of cold-formed lipped channel beams from LaBoube and Yu (1978a), and the new design rules developed based on them using the same approach used with LSBs. Finite element models of LSBs in shear were also developed to investigate the ultimate shear strength behaviour of LSBs including their elastic and post-buckling characteristics. They were validated by comparing their results with experimental test results. Details of the finite element models of LSBs, the nonlinear analysis results and their comparisons with experimental results are presented in this thesis. Finite element analysis results showed that the current cold-formed steel design rules are very conservative for the shear design of LSBs. They also confirmed other experimental findings relating to elastic and post-buckling shear strength of LSBs. A detailed parametric study based on validated experimental finite element model was undertaken to develop an extensive shear strength data base and was then used to confirm the accuracy of the new shear strength equations proposed in this thesis. Experimental and numerical studies were also undertaken to investigate the shear behaviour of LSBs with web openings. Twenty six shear tests were first undertaken using a three point loading arrangement. It was found that AS/NZS 4600 and Shan et al.'s (1997) design equations are conservative for the shear design of LSBs with web openings while McMahon et al.'s (2008) design equation are unconservative. Experimental finite element models of LSBs with web openings were then developed and validated by comparing their results with experimental test results. The developed nonlinear finite element model was found to predict the shear capacity of LSBs with web opening with very good accuracy. Improved design equations have been proposed for the shear capacity of LSBs with web openings based on both experimental and FEA parametric study results. This thesis presents the details of experimental and numerical studies of the shear behaviour and strength of LSBs with and without web openings and the results including the developed accurate design rules.