19 resultados para Immunomagnetic Separation

em University of Queensland eSpace - Australia


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Aims: A survey to determine the prevalence and numbers of Salmonella in beef cattle presented for slaughter at abattoirs across Australia was conducted between September 2002 and January 2003. Methods and Results: Automated immunomagnetic separation (AIMS) was used for detection and isolation of Salmonella enriched from cattle faeces. Salmonella were enumerated from positive samples using a combination of the Most Probable Number (MPN) technique and AIMS. A total of 310 faecal samples were tested, 155 were from lot-fed cattle and 155 from grass-fed cattle. Salmonella spp. were isolated from 21 (6.8%) of the cattle and the prevalence amongst grass-fed cattle (4.5%) was not significantly different to that found in lot-fed cattle (9%). Counts of Salmonella in positive faeces varied from

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Aims: To determine the prevalence and concentration of Escherichia coli O157 shed in faeces at slaughter, by beef cattle from different production systems. Methods and Results: Faecal samples were collected from grass-fed (pasture) and lot-fed (feedlot) cattle at slaughter and tested for the presence of E. coli O157 using automated immunomagnetic separation (AIMS). Escherichia coli O157 was enumerated in positive samples using the most probable number (MPN) technique and AIMS and total E. coli were enumerated using Petrifilm. A total of 310 faecal samples were tested (155 from each group). The geometric mean count of total E. coli was 5 x 10(5) and 2.5 x 10(5) CFU g(-1) for lot- and grass-fed cattle, respectively. Escherichia coli O157 was isolated from 13% of faeces with no significant difference between grass-fed (10%) and lot-fed cattle (15%). The numbers of E. coli O157 in cattle faeces varied from undetectable (

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Fecal culture for Escherichia coli 0157:H7 was compared to rectoanal mucosal swab (RAMS) culture in dairy heifers over a 1-year period. RAMS enrichment culture was as sensitive as fecal culture using immunomagnetic separation (IMS) (P = 0.98, as determined by a chi-square test). RAMS culture is less costly than fecal IMS culture and can yield quantitative data.

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The two sets of connected membranes induced in Kunjin virus-infected cells are characterized by the presence of NS3 helicase/protease in both, and by RNA-dependent RNA polymerase (RdRp) activity plus the associated double-stranded RNA (dsRNA) template in vesicle packets (VP), or by the absence of both the VP-specific markers in the convoluted membranes/paracrystalline arrays (CM/PC). Attempts were made to separate flavivirus-induced membranes by sedimentation or flotation analyses in density gradients of sucrose or iodixanol, respectively, after treatment of cell lysates by sonication, osmotic shock, or tryptic digestion. Only osmotic shock treatment provided suggestive evidence of separation. This was explored by flow cytometry analysis (FCA) of RdRp active membrane fractions from a sucrose gradient, using dual fluorescent labelling via antibodies to NS3 and dsRNA. FCA revealed the presence of a dual labelled membrane population indicative of VP, and in a faster sedimenting fraction a membrane population able to be labelled only in NS3, representative of CM/PC and associated (R)ER. It was postulated that osmotic shock ruptured the bounding membrane of the VP, releasing the enclosed small vesicles associated with the Kunjin virus replication complex characterized previously. Notably, the presence of the full spectrum of nonstructural proteins in some membrane fractions was not a reliable marker for RdRp activity. These experiments may provide the opportunity for isolation of relatively pure flavivirus replication complexes in their native membrane-associated state by fluorescence-activated cell sorting. (C) 2004 Elsevier B.V. All rights reserved.

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Reasons for performing study: Acute laminitis is characterised by hoof lamellar dermal-epidermal separation at the basement membrane (BM) zone. Hoof lamellar explants cultured in vitro can also be made to separate at the basement membrane zone and investigating how this occurs may give insight into the poorly understood pathophysiology of laminitis. Objectives: To investigate why glucose deprivation and metalloproteinase (MMP) activation in cultured lamellar explants leads to dermo-epidermal separation. Methods: Explants, cultured without glucose or with the MMP activator p-amino-phenol-mercuric acetate (APMA), were subjected to tension and processed for transmission electron microscopy (TEM). Results: Without glucose, or with APMA, explants under tension separated at the dermo-epidermal junction. This in vitro separation occurred via 2 different ultrastructural processes. Lack of glucose reduced hemidesmosomes (HDs) numbers until they disappeared and the basal cell cytoskeleton collapsed. Anchoring filaments (AFs), connecting the basal cell plasmalemma to the BM, were unaffected although they failed under tension. APMA activation of constituent lamellar MMPs did not affect HDs but caused AFs to disappear, also leading to dermo-epidermal separation under tension. Conclusions: Natural laminitis may occur in situations where glucose uptake by lamellar basal cells is compromised (e.g. equine Cushing's disease, obesity, hyperlipaemia, ischaemia and septicaemia) or when lamellar MMPs are activated (alimentary carbohydrate overload). Potential relevance: Therapies designed to facilitate peripheral glucose uptake and inhibit lamellar MMP activation may prevent or ameliorate laminitis.

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Miscibility and phase separation in the blends of phenolphthalein poly(aryl ether ketone) (PPAEK) and poly(ethylene oxide) (PEO) were investigated by means of differential scanning calorimetry (DSC). The PPAEK/PEO blends prepared by solution casting from N,N-dimethylformamide (DMF) displayed single composition-dependent glass transition temperatures (T-g), intermediate between those of the pure components, suggesting that the blend system is miscible in the amorphous state at all compositions. All the blends underwent phase separation at higher temperatures and the system exhibited a lower critical solution temperature (LCST) behavior. A step-heating thermal analysis was designed to determine the phase boundaries with DSC. The significant changes in the thermal properties of blends were utilized to judge the mixing status for the blends and the phase diagram was thus established. (C) 2004 Elsevier B.V. All rights reserved.

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Quantum optics experiments on bright beams are based on the spectral analysis of field fluctuations and typically probe correlations between radio-frequency sideband modes. However, the extra degree of freedom represented by this dual-mode picture is generally ignored. We demonstrate the experimental operation of a device which can be used to separate the quantum sidebands of an optical field. We use this device to explicitly demonstrate the quantum entanglement between the sidebands of a squeezed beam.

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Marriage breakdown through separation and divorce is a pervasive feature of Australian society. But little research investigates the social factors associated with marital breakdown in Australia. This study builds on and extends Australian research by using survival analysis models to examine patterns of association among temporal, life-course, attitudinal and economic factors associated with marital breakdown. Using data from the Household Income and Labour Dynamics in Australia (HILDA) survey, we find marital breakdown in Australia is socially patterned in similar ways to other Western countries. But our findings point to several directions for future research into marriage breakdown in Australia, and we identify certain unique features of Australian marriage breakdown that warrant a more detailed investigation, such as the relationship between ethnic origin and the risk of marital breakdown.

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Criticism of religiously motivated contributions to public policy debate is largely misconceived. It assumes that the mischief which constitutional separation of church and state is supposed to cure is a domination of the state by the church. This presents only one side of the story. Subservience by the church to the slate should also be avoided. The law of a liberal state is legitimate to the extent that it does not conflict with the basic moral values of its citizens. Therefore, an ongoing conversation about basic values is necessary. Allowing churches and individual believers the freedom to make distinctive 'religious' contributions to this conversation is consistent with the separation of church and state. It is an aspect of the liberal democratic state's obligation to listen to all perspectives on difficult moral issues. A close relationship between church and state, on the other hand, has the capacity to impede the conversation.