Mechanism and Function of Actin Pedestal Formation by Enterohemorrhagic \u3cem\u3eEscherichia coli\u3c/em\u3e O157:H7: A Dissertation

Enterohemorrhagic Escherichia coli O157:H7 (EHEC) and enteropathogenic E. coli O127:H7 (EPEC) induce characteristic F-actin rich pedestals on infected mammalian cells. Each pathogen delivers its own translocated intimin receptor (Tir) to the host cell to act as a receptor for the bacterial outer membrane adhesin, intimin. Interaction of translocated Tir with intimin is essential for mammalian cell binding and host colonization, as well as to induce actin pedestal formation in vitro. In spite of these parallels, EHEC and EPEC Tir appear to generate actin pedestals by distinct mechanisms. Further, while the ability to form actin pedestals is a striking phenotype, the function of pedestals during infection remains unclear. To address these issues, a systematic and quantitative analysis of Tir-mediated actin assembly was conducted. We identified a three-residue Tir sequence involved in actin pedestal formation for both EHEC and EPEC, and developed evidence that the two pathogens trigger a common pathway for actin assembly. Further, the ability of these bacteria to promote actin assembly appears to promote both intimin-mediated bacterial binding in vitro and optimal colonization during experimental animal infection

Shared Approach to Performance Assessment: Final Report on the Demonstration Projects

The aim of the overall project is:- To explore and document the ways in which professionals from disciplines other than social work can contribute to the performance assessment of social work students. It was agreed by the project team that we should also be exploring the converse of this, where other professionals may be less able to contribute to the shared assessment. The earlier practice audit and literature review, available on the Institute website, identified a number of themes that have formed the backdrop for the demonstration projects. These included:- • Shared assessment already takes place on social work courses, for example law and social sciences, and that in itself is not contentious. • There is a difference between sharing subject teaching and learning for collaborative practice. • The literature shows that to date, the preoccupation of educators has been with the what and how of teaching, as opposed to assessment. • Shared learning and assessment took place in a range of forms and sizes. As a general rule the larger the student groups the more complex the planning and management. However, small scale examples showed that successful outcomes could be achieved for less work. • Assessments ranged from the traditional essay and case studies to presentations and community profiling. • More was known about University based units and very little about assessment on practice placements. Therefore, we needed to focus on the following question in the demonstrations: What is complicated and contentious about shared assessment

Comparison of rapid laboratory tests for failure of passive transfer in the bovine

peer-reviewedBackground Failure of passive transfer of maternal immunity via colostrum can occur in the bovine, and a number of blood tests have been developed to test calves for this failure. It is not clear which test is most suitable for this purpose. The objective was to examine the most commonly used tests for failure of passive transfer and to decide which is most suitable for routine laboratory use. 126 serum samples were taken from calves of dairy cows after birth but prior to colostrum feeding, and at 48 h of age. Five different tests were compared against radial immunodiffusion which is considered the appropriate reference method. These tests were serum gamma-glutamyltransferase levels, serum protein levels, serum globulin levels, an enzyme linked immunosorbent assay and the zinc sulphate turbidity test. Results The tests examined displayed high sensitivity but widely varying specificity. Examination of the use of different cut-off points allowed some improvement in specificity at the expense of sensitivity, but the tests which had performed best at the original cut-off points still displayed the best performance. Gamma-glutamyltransferase levels as a measure of colostrum absorption returned, in this study, the best balance between sensitivity and specificity. The ELISA used in this study and serum globulin levels displayed performance similar to the gamma-glutamyltransferase levels. Serum total protein was less successful than others examined at providing both sensitivity and specificity but may, when performed via refractometer, be useful for on-farm testing. As currently performed the poor sensitivity for which the zinc sulphate turbidity test is most often criticized is evident. Modification of the cut-off point to increase specificity is less successful at balancing these parameters than the ELISA, gamma-glutamyltransferase levels, and globulin levels. Conclusions Gamma-glutamyltransferase levels, ELISA testing and circulating globulin levels performed best in detecting failure of passive transfer in serum samples, although all three had some practical considerations

Test Device and Process for Assessment of Smithfield Facility Aging

● Establishing structural objective tests and recommendations for hog production facilities. ● The risks are to the livestock, growers, and Smithfield for potential collapses of hog facilities in NW Iowa. ● We assess our solution to be applicable to a large portion of the hog production industry to quickly and objectively measure truss deterioration and provide detailed monetary and structural guidance to growers

Developing Objective Tests and Repair Guidelines for Hog Facility Trusses

• Predicting naturally ventilated hog barns life expectancy • Increase useful life of facilities and provide options to growers • Barn collapses and safety of personnel, livestock, facilities, and other assets • Provide cost breakdown of complete replacement vs. truss repair option

Alumina-Forming Austenitics: A New Approach to Thermal and Degradation Resistant Stainless Steels for Industrial Use

A series of developmental AFA alloys was selected for study based on: 25 Ni wt.% (alloys A-F), 20 wt% Ni (alloys G-H), and 12 Ni wt.% (alloys I-L). An emphasis in this work was placed on the lower alloy content direction for AFA alloys to reduce alloy raw material cost, rather than more highly alloyed and costly AFA alloys for higher temperature performance. Alloys A-D explored the effects of Al (3-4 wt.%) and C (0.05-0.2 wt.%) in the Fe-25Ni-14Cr-2Mn-2Mo-1W-1Nb wt.% base range; alloys E and F explored the effects of removing costly Mo and W additions in a Fe-25Ni-14Cr-4Al-2.5Nb-2Mn-0.2C base, alloys G and H examined Nb (1-2.5wt.%) and removal of Mo, W in a Fe-20Ni-14Cr-3Al-2Mn-0.2 C wt.% base; and alloys I-L examined effects of C (0.1-0.2 wt.%) and Mn (5-10 wt.%) on a low cost Fe-14Cr-12Ni-3Cu-2.5Al wt.% base (no Mo, W additions). Creep testing resulted in elemental trends that included the beneficial effect of higher carbon and lower niobium in 20-25%Ni AFA alloys and, the beneficial of lower Mn in 12%Ni AFA alloys. Corrosion tests in steam and sulfidation-oxidation environments showed, in general, these alloys were capable of a ten-fold improvement in performance when compared to conventional austenitic stainless steels. Also, corrosion test results in metal-dusting environments were promising and, warrant further investigation

Enhanced Actin Pedestal Formation by Enterohemorrhagic Escherichia coli O157:H7 Adapted to the Mammalian Host

Upon intestinal colonization, enterohemorrhagic Escherichia coli (EHEC) induces epithelial cells to generate actin “pedestals” beneath bound bacteria, lesions that promote colonization. To induce pedestals, EHEC utilizes a type III secretion system to translocate into the mammalian cell bacterial effectors such as translocated intimin receptor (Tir), which localizes in the mammalian cell membrane and functions as a receptor for the bacterial outer membrane protein intimin. Whereas EHEC triggers efficient pedestal formation during mammalian infection, EHEC cultured in vitro induces pedestals on cell monolayers with relatively low efficiency. To determine whether growth within the mammalian host enhances EHEC pedestal formation, we compared in vitro-cultivated bacteria with EHEC directly isolated from infected piglets. Mammalian adaptation by EHEC was associated with a dramatic increase in the efficiency of cell attachment and pedestal formation. The amounts of intimin and Tir were significantly higher in host-adapted than in in vitro-cultivated bacteria, but increasing intimin or Tir expression, or artificially increasing the level of bacterial attachment to mammalian cells, did not enhance pedestal formation by in vitro-cultivated EHEC. Instead, a functional assay suggested that host-adapted EHEC translocate Tir much more efficiently than does in vitro-cultivated bacteria. These data suggest that adaptation of EHEC to the mammalian intestine enhances bacterial cell attachment, expression of intimin and Tir, and translocation of effectors that promote actin signaling