Microbial source tracking (MST) results, obtained using identical sample sets and pulsed field gel electrophoresis (PFGE), repetitive element PCR (rep-PCR) and ribotyping techniques were compared. These methods were performed by six investigators in analysis of duplicate, blind sets of water samples spiked with feces from five possible sources (sewage, human, dog, cow and seagull). Investigators were provided with samples of the fecal material used to inoculate the water samples for host origin database construction. All methods correctly identified the dominant source in the majority of the samples. Modifications of some of these methods correctly identified the dominant sources in over 90% of the samples; however, false positive rates were as high as 57%. The high false positive rates appeared to be indirectly proportional to the levels of stringency applied in pattern analysis. All the methods produced useful data but the results highlighted the need to modify and optimize these methods in order to minimize sources of error.
Amyloid β (Aβ) is thought to promote neuronal cell loss in Alzheimer's disease (AD), in part through the generation of reactive oxygen species (ROS) and subsequent activation of mitogenactivated protein kinase (MAPK) pathways. Protein phosphatase 5 (PP5) is a ubiquitously expressed serine/threonine phosphatase which has been implicated in several cell stress response pathways and shown to inactivate MAPK pathways through key dephosphorylation events. Therefore we examined whether PP5 protects dissociated embryonic rat cortical neurons in vitro from cell death evoked by Aβ. As predicted, neurons in which PP5 expression was decreased by siRNA treatment were more susceptible to Aβ toxicity. In contrast, overexpression of PP5, but not the inactive PP5 mutant, H304Q, prevented MAPK phosphorylation and neurotoxicity induced by Aβ. PP5 also prevented cell death caused by direct treatment with H 2 O 2 , but did not prevent Aβ-induced production of ROS. Thus, the neuroprotective effect of PP5 requires its phosphatase activity and lies downstream of Aβ-induced generation of ROS. In summary, our data indicate that PP5 plays a pivotal neuroprotective role against cell death induced by Aβ and oxidative stress. Consequently, PP5 might be an effective therapeutic target in AD and other neurodegenerative disorders in which oxidative stress is implicated.
Frequent outbreaks of foodborne illness have been increasing the need for simple, rapid and
sensitive methods to detect foodborne pathogens. Conventional methods for pathogen detection and
identification are labor-intensive and take days to complete. Some immunological rapid assays are
developed, but these assays still require prolonged enrichment steps. Biosensors have shown great
potential for the rapid detection of foodborne pathogens. Among the biosensors, fiber-optic methods
have much potential because they can be very sensitive and simple to operate. Fiber-optic biosensors
typically use a light transmittable, tapered fiber to send excitation laser light to the detection surface
and receive emitted fluorescent light. The fluorescent light excited by an evanescent wave generated
by the laser is quantitatively related to fluorophor-labeled biomolecules immobilized on the fiber
surface. A portable and automated fiber-optic biosensor, RAPTOR (Research International, Monroe,
WA), was used to detect Salmonella enteritidis in food samples. A binding inhibition assay based on
the biosensor was developed to detect the bacteria in hot dog samples. The biosensor and the binding
inhibition assay could detect 104 cfu/ml of bacteria in less than 10 min of assay time.
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