993 resultados para Transferência lateral de genes


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The flexural capacity of of a new cold-formed hollow flange channel section known as LiteSteel beam (LSB) is limited by lateral distortional buckling for intermediate spans, which is characterised by simultaneous lateral deflection, twist and web distortion. Recent research has developed suitable design rules for the member capacity of LSBs. However, they are limited to a uniform moment distribution that rarely exists in practice. Many steel design codes have adopted equivalent uniform moment distribution factors to accommodate the effect of non-uniform moment distributions in design. But they were derived mostly based on the data for conventional hot-rolled, doubly symmetric I-beams subject to lateral torsional buckling. The effect of moment distribution for LSBs, and the suitability of the current steel design code rules to include this effect for LSBs are not yet known. This paper presents the details of a research study based on finite element analyses of the lateral buckling strength of simply supported LSBs subject to moment gradient effects. It also presents the details of a number of LSB lateral buckling experiments undertaken to validate the results of finite element analyses. Finally, it discusses the suitability of the current design methods, and provides design recommendations for simply supported LSBs subject to moment gradient effects.

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Interferometry is a sensitive technique for recording tear film surface irregularities in a noninvasive manner. At the same time, the technique is hindered by natural eye movements resulting in measurement noise. Estimating tear film surface quality from interferograms can be reduced to a spatial-average-localized weighted estimate of the first harmonic of the interference fringes. However, previously reported estimation techniques proved to perform poorly in cases where the pattern fringes were significantly disturbed. This can occur in cases of measuring tear film surface quality on a contact lens on the eye or in a dry eye. We present a new estimation technique for extracting the first harmonic from the interference fringes that combines the traditional spectral estimation techniques with morphological image processing techniques. The proposed technique proves to be more robust to changes in interference fringes caused by natural eye movements and the degree of dryness of the contact lens and corneal surfaces than its predecessors, resulting in tear film surface quality estimates that are less noisy

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The purpose of this paper is to determine the prevalence of the toxic shock toxin gene (tst) and to enumerate the circulating strains of methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) in Australian isolates collected over two decades. The aim was to subtype these strains using the binary genes pvl, cna, sdrE, pUB110 and pT181. Isolates were assayed using real-time polymerase chain reaction (PCR) for mecA, nuc, 16 S rRNA, eight single-nucleotide polymorphisms (SNPs) and for five binary genes. Two realtime PCR assays were developed for tst. The 90 MRSA isolates belonged to CC239 (39 in 1989, 38 in 1996 and ten in 2003), CC1 (two in 2003) and CC22 (one in 2003). The majority of the 210 MSSA isolates belonged to CC1 (26), CC5 (24) and CC78 (23). Only 18 isolates were tst-positive and only 15 were pvl-positive. Nine MSSA isolates belonged to five binary types of ST93, including two pvlpositive types. The proportion of tst-positive and pvl-positive isolates was low and no significant increase was demonstrated. Dominant MSSA clonal complexes were similar to those seen elsewhere, with the exception of CC78. CC239 MRSA (AUS-2/3) was the predominant MRSA but decreased significantly in prevalence, while CC22 (EMRSA-15) and CC1 (WA-1) emerged. Genetically diverse ST93 MSSA predated the emergence of ST93- MRSA (the Queensland clone).

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The LiteSteel Beam (LSB) is a new hollow flange section developed by OneSteel Australian Tube Mills using their patented dual electric resistance welding and automated continuous roll-forming technologies. It has a unique geometry consisting of torsionally rigid rectangular hollow flanges and a relatively slender web. It has found increasing popularity in residential, industrial and commercial buildings as flexural members. The LSB is considerably lighter than traditional hot-rolled steel beams and provides both structural and construction efficiencies. However, the LSB flexural members are subjected to a relatively new lateral distortional buckling mode, which reduces their member moment capacities. Unlike the commonly observed lateral torsional buckling of steel beams, the lateral distortional buckling of LSBs is characterised by simultaneous lateral defection, twist and cross sectional change due to web distortion. The current design rules in AS/NZS 4600 (SA, 2005) for flexural members subject to lateral distortional buckling were found to be conservative by about 8% in the inelastic buckling region. Therefore, a new design rule was developed for LSBs subject to lateral distortional buckling based on finite element analyses of LSBs. The effect of section geometry was then considered and several geometrical parameters were used to develop an advanced set of design rules. This paper presents the details of the finite element analyses and the design curve development for hollow flange sections subject to lateral distortional buckling.

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A new cold-formed steel beam, known as the LiteSteel Beam (LSB), has the potential to transform the low-rise building industry. The new beam is effectively a channel section with two rectangular hollow flanges and a slender web, and is manufactured using a simultaneous cold-forming and electric resistance welding process. Research into the flexural behaviour of single LSB members showed that the LSBs are susceptible to lateral distortional buckling effects and their moment capacities are significantly reduced for intermediate spans. Build-up LSB sections are expected to improve their flexural capacity and to enhance their applications. They are also likely to mitigate the detrimental effects of lateral distortional buckling observed with single LSB members of intermediate spans. However, the behaviour of build up beams is not well understood. Currently available design rules were found to be inadequate to predict the member moment capacities of back to back LSBs. Therefore a research project based on both experimental and numerical studies was undertaken to investigate the flexural behaviour of back to back LSBs with various longitudinal connection spacings under a uniform moment. New design rules were developed using the moment capacity data obtained using finite element analyses and experimental tests. This paper presents the details of the development of design rules for the back to back LSB sections.

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Cold-formed steel beams are increasingly used as floor joists and bearers in residential, industrial and commercial buildings. Their structural behaviour and moment capacities are influenced by lateral-torsional buckling and hence a research study was undertaken to investigate the lateral-torsional buckling behaviour of cold-formed steel lipped channel beams at ambient and elevated temperatures. For this purpose a finite element model of a simply supported cold-formed steel lipped channel beam under uniform bending was developed first and validated using available numberical and experimental results. It was then used in a detailed parametric study to simulate the lateral-torsional behaviour of cold-formed steel beams under varying conditions. The moment capacity results were then compared with the predictions from the current ambient temperature design rules in Australia, New Zealand, American and European codes for cold-formed steel structures. Some very interesting results have been obtained. European design rules are found to be conservative while Australian and American design rules are unsafe. This paper presents the results of finite element analyses for ambient temperature conditions, and the comparison with the current design rules.

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This project set out to investigate the behaviour of a pole frame house subjected to a lateral wind load. The behaviour of poles embedded in the ground was examined. The existing theoretical methods for determining lateral load capacity of an embedded pole were reviewed, and three common methods of pole embedment were tested at different depths to gauge the response of poles and types of pole embedment to a lateral load. The most suitable embedment method was used in the foundation for a full-scale model pole house, which was constructed and tested at various stages during the construction to examine the response of a pole house to lateral wind load. The full scale testing was also used to monitor the effect of the various structural components on the overall stiffuess of the house. The results from the full scale tests were used to calibrate a computer model of a pole house which could then be used to predict the behaviour of different configurations of pole house construction without the need for further expensive full scale tests.

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One approach to reducing the yield losses caused by banana viral diseases is the use of genetic engineering and pathogen-derived resistance strategies to generate resistant cultivars. The development of transgenic virus resistance requires an efficient banana transformation method, particularly for commercially important 'Cavendish' type cultivars such as 'Grand Nain'. Prior to this study, only two examples of the stable transformation of banana had been reported, both of which demonstrated the principle of transformation but did not characterise transgenic plants in terms of the efficiency at which individual transgenic lines were generated, relative activities of promoters in stably transformed plants, and the stability of transgene expression. The aim of this study was to develop more efficient transformation methods for banana, assess the activity of some commonly used and also novel promoters in stably transformed plants, and transform banana with genes that could potentially confer resistance to banana bunchy top nanovirus (BBTV) and banana bract mosaic potyvirus (BBrMV). A regeneration system using immature male flowers as the explant was established. The frequency of somatic embryogenesis in male flower explants was influenced by the season in which the inflorescences were harvested. Further, the media requirements of various banana cultivars in respect to the 2,4-D concentration in the initiation media also differed. Following the optimisation of these and other parameters, embryogenic cell suspensions of several banana (Musa spp.) cultivars including 'Grand Nain' (AAA), 'Williams' (AAA), 'SH-3362' (AA), 'Goldfinger' (AAAB) and 'Bluggoe' (ABB) were successfully generated. Highly efficient transformation methods were developed for both 'Bluggoe' and 'Grand Nain'; this is the first report of microprojectile bombardment transformation of the commercially important 'Grand Nain' cultivar. Following bombardment of embryogenic suspension cells, regeneration was monitored from single transfom1ed cells to whole plants using a reporter gene encoding the green fluorescent protein (gfp). Selection with kanamycin enabled the regeneration of a greater number of plants than with geneticin, while still preventing the regeneration of non-transformed plants. Southern hybridisation confirmed the neomycin phosphotransferase gene (npt II) was stably integrated into the banana genome and that multiple transgenic lines were derived from single bombardments. The activity, stability and tissue specificity of the cauliflower mosaic virus 358 (CaMV 35S) and maize polyubiquitin-1 (Ubi-1) promoters were examined. In stably transformed banana, the Ubi-1 promoter provided approximately six-fold higher p-glucuronidase (GUS) activity than the CaMV 35S promoter, and both promoters remained active in glasshouse grown plants for the six months they were observed. The intergenic regions ofBBTV DNA-I to -6 were isolated and fused to either the uidA (GUS) or gfjJ reporter genes to assess their promoter activities. BBTV promoter activity was detected in banana embryogenic cells using the gfp reporter gene. Promoters derived from BBTV DNA-4 and -5 generated the highest levels of transient activity, which were greater than that generated by the maize Ubi-1 promoter. In transgenic banana plants, the activity of the BBTV DNA-6 promoter (BT6.1) was restricted to the phloem of leaves and roots, stomata and root meristems. The activity of the BT6.1 promoter was enhanced by the inclusion of intron-containing fragments derived from the maize Ubi-1, rice Act-1, and sugarcane rbcS 5' untranslated regions in GUS reporter gene constructs. In transient assays in banana, the rice Act-1 and maize Ubi-1 introns provided the most significant enhancement, increasing expression levels 300-fold and 100-fold, respectively. The sugarcane rbcS intron increased expression about 10-fold. In stably transformed banana plants, the maize Ubi-1 intron enhanced BT6.1 promoter activity to levels similar to that of the CaMV 35S promoter, but did not appear to alter the tissue specificity of the promoter. Both 'Grand Nain' and 'Bluggoe' were transformed with constructs that could potentially confer resistance to BBTV and BBrMV, including constructs containing BBTV DNA-1 major and internal genes, BBTV DNA-5 gene, and the BBrMV coat protein-coding region all under the control of the Ubi-1 promoter, while the BT6 promoter was used to drive the npt II selectable marker gene. At least 30 transgenic lines containing each construct were identified and replicates of each line are currently being generated by micropropagation in preparation for virus challenge.

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Background The purpose of this study was to identify candidate metastasis suppressor genes from a mouse allograft model of prostate cancer (NE-10). This allograft model originally developed metastases by twelve weeks after implantation in male athymic nude mice, but lost the ability to metastasize after a number of in vivo passages. We performed high resolution array comparative genomic hybridization on the metastasizing and non-metastasizing allografts to identify chromosome imbalances that differed between the two groups of tumors. Results This analysis uncovered a deletion on chromosome 2 that differed between the metastasizing and non-metastasizing tumors. Bioinformatics filters were employed to mine this region of the genome for candidate metastasis suppressor genes. Of the 146 known genes that reside within the region of interest on mouse chromosome 2, four candidate metastasis suppressor genes (Slc27a2, Mall, Snrpb, and Rassf2) were identified. Quantitative expression analysis confirmed decreased expression of these genes in the metastasizing compared to non-metastasizing tumors. Conclusion This study presents combined genomics and bioinformatics approaches for identifying potential metastasis suppressor genes. The genes identified here are candidates for further studies to determine their functional role in inhibiting metastases in the NE-10 allograft model and human prostate cancer.

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Background The androgen receptor is a ligand-induced transcriptional factor, which plays an important role in normal development of the prostate as well as in the progression of prostate cancer to a hormone refractory state. We previously reported the identification of a novel AR coactivator protein, L-dopa decarboxylase (DDC), which can act at the cytoplasmic level to enhance AR activity. We have also shown that DDC is a neuroendocrine (NE) marker of prostate cancer and that its expression is increased after hormone-ablation therapy and progression to androgen independence. In the present study, we generated tetracycline-inducible LNCaP-DDC prostate cancer stable cells to identify DDC downstream target genes by oligonucleotide microarray analysis. Results Comparison of induced DDC overexpressing cells versus non-induced control cell lines revealed a number of changes in the expression of androgen-regulated transcripts encoding proteins with a variety of molecular functions, including signal transduction, binding and catalytic activities. There were a total of 35 differentially expressed genes, 25 up-regulated and 10 down-regulated, in the DDC overexpressing cell line. In particular, we found a well-known androgen induced gene, TMEPAI, which wasup-regulated in DDC overexpressing cells, supporting its known co-activation function. In addition, DDC also further augmented the transcriptional repression function of AR for a subset of androgen-repressed genes. Changes in cellular gene transcription detected by microarray analysis were confirmed for selected genes by quantitative real-time RT-PCR. Conclusion Taken together, our results provide evidence for linking DDC action with AR signaling, which may be important for orchestrating molecular changes responsible for prostate cancer progression.

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There are several noninvasive techniques for assessing the kinetics of tear film, but no comparative studies have been conducted to evaluate their efficacies. Our aim is to test and compare techniques based on high-speed videokeratoscopy (HSV), dynamic wavefront sensing (DWS), and lateral shearing interferometry (LSI). Algorithms are developed to estimate the tear film build-up time TBLD, and the average tear film surface quality in the stable phase of the interblink interval TFSQAv. Moderate but significant correlations are found between TBLD measured with LSI and DWS based on vertical coma (Pearson's r2=0.34, p<0.01) and higher order rms (r2=0.31, p<0.01), as well as between TFSQAv measured with LSI and HSV (r2=0.35, p<0.01), and between LSI and DWS based on the rms fit error (r2=0.40, p<0.01). No significant correlation is found between HSV and DWS. All three techniques estimate tear film build-up time to be below 2.5 sec, and they achieve a remarkably close median value of 0.7 sec. HSV appears to be the most precise method for measuring tear film surface quality. LSI appears to be the most sensitive method for analyzing tear film build-up.