75 resultados para indirect fluorescent antibody (IFA)


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The synthesis and complete characterisation of the fluorescent ligand, 4-acridinol-1-sulphonic acid (the acridine analogue of 8-quinolinol-5-sulfonic acid) is described. Using a judicious array of nuclear magnetic resonance spectroscopy experiments, the structural elucidation and full assignment of all proton and carbon chemical shifts were afforded. The 4-acridinol-1-sulphonic acid was found to behave in a similar manner to 8-quinolinol-5-sulphonic acid, forming fluorescent complexes with magnesium(II) and zinc(II). The uncorrected emission maxima for the metal–acridinol complexes were found to be at around 620 nm compared to 505 nm for the respective quinolinol complexes. Unfortunately, preliminary spectrofluorimetric analytical figures of merit revealed that the detection limits of the new acridinol metal complexes were one and a half orders of magnitude poorer than those attained with the corresponding quinolinol ligand. However, in contrast to 8-quinolinol-5-sulphonic acid, the 4-acridinol-1-sulphonic acid ligand showed considerable selectivity for magnesium(II) and zinc(II) over aluminium(III).

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Arabidopsis thaliana ecotype Columbia-0 was transformed with a green fluorescent protein (GFP) gene under control of a phenylalanine ammonia-lyase (PAL) promoter. PAL is a key enzyme of the phenylpropanoid pathway and is induced to high levels during plant stress. Constitutive expression of PAL1 promoter-controlled GFP occurred in vascular tissues within stems, leaves and roots and in developing flowers. PAL1 promoter–GFP expression was examined in leaves of transgenic plants subjected to an abiotic elicitor, mechanical wounding or to inoculation with the pathogens Pseudomonas syringae pv. tomato or Peronospora parasitica. Wounding of leaves and treatment with an abiotic elicitor and compatible interactions produced low to moderate levels of GFP. However, in incompatible interactions there were high levels of GFP produced. In incompatible interactions, the intensity of GFP fluorescence was similar to that produced in transgenic plants expressing GFP driven by the CaMV promoter. The bright green fluorescence produced in live cells and tissues was readily visualised using conventional fluorescence microscopy and was quantified using spectroflourometry. This is the first report of the use of GFP as a reporter of defence gene activation against pathogens. It has several advantages over other reporter genes including real time analysis of gene expression and visualisation of defence gene activation in a non-invasive manner.

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Individuals typically believe that they are less likely than the average person to experience negative events. Such “unrealistic optimism” (UO) has been assessed in two ways: directly (via a single question, requiring comparison of own risk to that of the average person) and indirectly (via separate questions about own risk and that of the average person). The study examined the equivalence of the direct and indirect measures of female students' UO for unwanted pregnancy. Participants (N = 120) answered questions about their own risk, that of the average female student, and their own risk relative to that of the average female student; responses and response times were recorded. There was only a moderate association between direct- and indirect-UO. Direct-UO was strongly associated with estimate of own risk, but only moderately associated with estimate of the average student's risk. Response times for the comparative risk and own risk questions did not differ, but participants took significantly longer to answer the question about average student's risk. The results suggest that the two measures of UO are not equivalent, but, rather, that individuals answering a comparative risk question focus mainly on their own risk, at the expense of that of the average person.

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Although initially Australia saw some high-profile successes in antidiscrimination cases for workers with family responsibilities, recent trends in appellate decisions such as Schou suggest that indirect discrimination concepts do not succeed for workers with family responsibilities. The limitations of an individual complaint model to address systemic disadvantage such as that experienced by ‘workercarers’ is simply in too much tension with entrenched expectations surrounding the contract of employment and the ‘ideal’ or ‘unencumbered’ worker. A re-imagining of the employment relationship and the role of the employer will be necessary to achieve substantive equality for workers with family responsibilities.

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The new charge neutral 4-amino-1,8-naphthalimide based anion sensors 2 and 3 bind to both acetate and dihydrogenphosphate with 1:1 stoichiometry through hydrogen bonding to both thiourea N–H atoms and in the case of dihydrogenphosphate, the naphthalimide 4 amino N–H group as well. This was clearly established from 1H NMR titration experiments with H2PO4- in DMSO-d6 where a substantial shift in the resonance for the naphthalimide N–H was observed concomitant with the expected migration of the thiourea N–H chemical shifts. The binding constants determined from the titration studies indicate that the new sensors bind H2PO4- more strongly than AcO. Fluorescence titrations with sensor 3 indicate quenching of 59% and 36% upon addition of acetate and dihydrogenphosphate, respectively.

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The article comments on the paper of Manfred Müller and Sandra Danielzik about the indirect approaches to obesity. They are looking for alternatives that are both insighful and well-grounded in experience of single or multiple behavioral interventions on obesity prevalence. The author suggested various approaches including community capacity, counter commercialism, cultural change and contagion creation.

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Australia is considered the driest populated continent in the world. Despite this, we consume the largest amount of water, per capita. While little of this water is used for the operation of buildings, buildings are now being designed to use less water. Additionally, rainwater collection and grey water recycling systems offer the potential to significantly reduce demand for fresh water. However, little is known about the water required directly and indirectly (ie., embodied in) construction materials and products. Embodied water comprises the water required directly for construction itself and the water consumed indirectly in the production and delivery of materials, products and services to construction. Water required directly for construction is likely to be insignificant compared to the indirect water required for the manufacture of construction materials and products (ie., through materials and other products required to support construction). There is currently a lack of research into embodied water requirements by the construction sector. The relationship between the embodied water and the operational water is also unknown, apart from a handful of studies based solely on national average statistics known as 'input-output' data. The aim of this paper is therefore to model the water required directly and indirectly by construction, integrating currently available public domain industry data with input-output data. The coverage of the industry data relative to the input-output data was evaluated for a typical commercial building, and was found to be very low.

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This study examined the influence of trait resilience and trait anxiety on self-esteem and investigated the mediating role of positive and negative affect in this relationship. Specifically, it was proposed that trait resilience and trait anxiety may exert indirect effects on self-esteem by promoting increased positive and negative affect, respectively. The final sample comprised of 240 participants (age, M = 21.55, SD = 4.16) who completed questionnaire measures of trait resilience (CD-RISC; Connor & Davidson, 2003), trait anxiety (STAI-T; Spielberger, Gorsuch, Lushene, Vagg, & Jacobs, 1983), affect (PANAS; Watson, Clark, & Tellegen, 1988) and self-esteem (RSES; Rosenberg, 1965). Path analyses (AMOS), controlling for measurement error revealed significant indirect effects of trait anxiety and trait resilience on self-esteem via negative and positive affect. The indirect model provided a very close fit to the data; estimation of the full model (direct paths) did not yield a significantly better fit. It was concluded that the impact of trait anxiety and trait resilience on self-esteem may be due to their effects on regulating affective experiences which in turn may be more proximal predictors of individual feelings of self-worth.

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Water consumed directly by the construction industry is known to be of little importance. However, water consumed in the manufacture of goods and services required by construction may be significant in the context of a building's life cycle water requirements and the national water budget. This paper evaluates the significance of water embodied in the construction of individual buildings. To do this, an input-output-based hybrid embodied water analysis was undertaken on 17 Australian non-residential case studies. It was found that there is a considerable amount of water embodied in construction. The highest value was 20.1 kilolitres (kL)/m2 gross floor area (GFA), representing many times the enclosed volume of the building, and many years worth of operational water. The water required by the main construction process is minimal. However, the water embodied in building materials is considerable. These findings suggest that the selection of elements and materials has a great impact on a building's embodied water. This research allows the construction industry to evaluate design and construction in broad environmental terms to select options that might be cost neutral or possibly cost positive while retaining their environmental integrity. The research suggests policies focused on operational water consumption alone are inadequate.

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This mini review highlights the synthesis and photophysical evaluation of anion sensors, for nonaqueous solutions, that have been developed in our laboratories over the last few years. We have focused our research mainly on developing fluorescent photoinduced electron transfer (PET) sensors based on the fluorophore-spacer-anion receptor principle using several anthracene (emitting in the blue) and 1,8-naphthalimide (emitting in the green) fluorophores, with the aim of targeting biologically and industrially relevant anions such as acetates, phosphate and amino acids, as well as halides such as fluoride. The receptors and the fluorophore are separated by a short methyl or ethyl spacer, where the charge neutral anion receptors are either aliphatic or aromatic urea (or thiourea) moieties. For these, the anion recognition is through hydrogen bonding, yielding anion:receptor complexes. Such bonding gives rise to enhanced reduction potential in the receptor moieties which causes enhancement in the rate of PET quenching of the fluorophore excited state from the anion:receptor moiety. This design can be further elaborated on by incorporating either two fluorophores, or urea/thiourea receptors into the sensor structures, using anthracene as a fluorophore. For the latter design, the sensors were designed to achieve sensing of bis-anions, such as di-carboxylates or pyrophosphate, where the anion bridged the anthracene moiety. In the case of the naphthalimide based mono-receptor based PET sensors, it was discovered that in DMSO the sensors were also susceptible to deprotonation by anions such as F− at high concentrations. This led to substantial changes in the absorption spectra of these sensors, where the solution changed colour from yellow/green to deep blue, which was clearly visible to the naked eye. Hence, some of the examples presented can act as dual fluorescent-colorimetric sensors for anions. Further investigations into this phenomenon led to the development of simple colorimetric sensors for fluorides, which upon exposure to air, were shown to fix carbon dioxide as bicarbonate.

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This paper presents a system that employs random forests to formulate a method for subcellular localisation of proteins. A random forest is an ensemble learner that grows classification trees. Each tree produces a classification decision, and an integrated output is calculated. The system classifies the protein-localisation patterns within fluorescent microscope images. 2D images of HeLa cells that include all major classes of subcellular structures, and the associated feature set are used. The performance of the developed system is compared against that of the support vector machine and decision tree approaches. Three experiments are performed to study the influence of the training and test set size on the performance of the examined methods. The calculated classification errors and execution times are presented and discussed. The lowest classification error (2.9%) has been produced by the developed system.

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The visual systems of birds are hypothesized to have higher temporal resolution than those of humans, suggesting that they may be able to perceive the flicker emitted from conventional low-frequency fluorescent lights (LF; 100 Hz in Europe, 120 Hz in the U.S.A.). These lights are commonly used in the housing of captive birds and this may affect both their welfare and performance in experiments. We carried out mate choice experiments on European starlings, Sturnus vulgaris, under both low- and high-frequency fluorescent lights (HF; > 30 kHz, at which flicker is imperceptible). Indicators of male condition and size, together with the reflectance spectra and length of the males' throat feathers, were also recorded to ascertain which variables correlated with female preference. Females ranked males consistently under HF, but not LF, lighting, and individual females chose different males under the two lighting types. Under HF lighting, females chose to spend more time with males that had longer throat feathers. The flicker rate of the light clearly affected the choices made by the females, possibly because of nonspecific stress effects or decreased discrimination ability. Our results imply that careful interpretation of mate choice experiments is needed, especially with regard to the lighting types used, to elucidate the real cause behind any variation shown. (c) 2006 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.