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Biomedical subjects

J Keen

Publications and source records attributed to J Keen.

At least 37 records · Page 2Linked to original sources
PubMed1999

Evaluation of a hospital picture archiving and communication system.

OBJECTIVES: To establish the net costs to the hospital and the broad range of benefits associated with a hospital-wide picture archiving and communication system (PACS) that comprised digital acquisition, storage and transmission of radiological images via a hospital-wide network to 150 workstations. METHODS: 'Before and after' comparisons and time series analyses at Hammersmith Hospital (London, UK), and comparison with five other British hospitals where PACS was not being installed. The cost analysis considered implementation costs and changes in key elements of hospital running costs, including the impact of changes in the length of inpatient stays. A range of benefit measures were investigated, including image availability, avoidance of repeat imaging, avoidance of exposure to radiation, patient turn-round speed, time from examination to image availability in intensive care, avoidance of diagnostic 'errors' by casualty doctors, the additional diagnostic value of PACS-based images and clinician satisfaction. RESULTS: The annual equivalent capital cost of the PACS was 1.7 million Pounds (annual equivalent replacement cost: 0.8 million Pound). Overall, the PACS substantially increased running costs. No convincing evidence of a PACS-induced change in length of inpatient stay was found. PACS was associated with some improvements in the performance of the radiology department: improved image availability (97.7% versus 86.9%), lower repeat imaging rate (7.3% versus 9.9%) and 20% lower total radiation doses for examinations of the lateral lumbar spine. No improvements were identified in the quality of the radiology reporting service. Benefits outside radiology included shorter time from examination to image availability for routine uses in intensive care (19 versus 37 minutes), and a lower rate of diagnostic 'errors' in casualty (0.65% versus 1.51%). High levels of satisfaction with PACS were found amongst both providers and clinical users. CONCLUSIONS: PACS was almost universally preferred by users and brought many operational and clinical benefits. However, these advantages came at a significant capital and net running cost.

Cost-Benefit Analysis↗
PubMed1998

Evidence for the existence of a novel pregnancy-associated soluble variant of the vascular endothelial growth factor receptor, Flt-1.

Angiogenesis is essential in physiological processes including ovulation, implantation and pregnancy. One of the most potent regulators is the cytokine vascular endothelial growth factor (VEGF). We provide evidence for a novel pregnancy-associated soluble variant of the VEGF receptor Flt-1. VEGF ranged from undetectable to 157.3 pg/ml (mean 49.9 pg/ml, SD 48.4 pg/ml) in plasma samples from normal volunteers (n = 10), but was undetectable in plasma from pregnant women (n = 12) and amniotic fluid (n = 10). Recoveries of spiked VEGF were poor in pregnancy-related samples, indicating the presence of VEGF-binding activity which was confirmed using biosensor and chromatographic techniques. Partial purification and protein sequencing indicated a novel soluble form of Flt-1 with a subunit size of 150 kDa. Normally present as a multimeric structure of approximately 400-550 kDa, complexes of 600-700 kDa were formed following binding of multiple VEGF molecules. Reverse transcriptase polymerase chain reaction of Flt-1 in placenta, amnion, chorion, human umbilical vein endothelial cells and cord blood samples produced bands of the predicted sizes but failed to identify any additional RNA species, and possible reasons for this are discussed. Soluble Flt-1 may be important in regulating the actions of VEGF in angiogenesis and trophoblast invasion and may have therapeutic implications in diseases with inappropriate angiogenesis such as proliferative retinopathies and cancer.

Amino Acid Sequence↗
PubMed1998

Development of a blocking enzyme-linked immunosorbent assay for detection of serum antibodies to O157 antigen of Escherichia coli.

The O157 antigen of Escherichia coli shares structural elements with lipopolysaccharide (LPS) antigens of other bacterial species, notably Brucella abortus and Yersinia enterocolitica 09, a fact that confounds the interpretation of assays for anti-O157 antibodies. To address this problem, a blocking enzyme-linked immunosorbent assay (bELISA) was designed with E. coli O157:H7 LPS as the antigen and a monoclonal antibody specific for E. coli O157, designated 13B3, as the competing antibody. The bELISA had equivalent sensitivity to, and significantly higher specificity than, the indirect ELISA (iELISA), detecting anti-O157 antibodies in sera from cattle experimentally inoculated with O157:H7. Only 13% of sera from naive heifers vaccinated for or experimentally infected with B. abortus had increased anti-O157 bELISA titers, while 61% of anti-O157 iELISA titers were increased. The bELISA is a sensitive and specific method for the detection of serum antibodies resulting from exposure to E. coli O157.

Animals↗
PubMed1998

Radiology report times: impact of picture archiving and communication systems.

OBJECTIVE: We investigated the impact on radiologist reporting time of the change from conventional film to hard-copy computerized radiography and of the subsequent move to soft-copy images on picture archiving and communication system (PACS) workstations. MATERIALS AND METHODS: A controlled before and after research design was undertaken. Data were collected on four occasions: two relating to conventional film, one relating to hard-copy computerized radiography, and one relating to soft-copy PACS images. Data collection was by direct observation of radiology reporting sessions by independent health service researchers. Data were collected on report times, details of images viewed, characteristics of the radiologist, and details of interruptions. To control for potential biases in the before and after comparisons, ordinary least squares multiple regression analysis was used. The principal comparison was between reports with PACS and reports with computerized radiography hard-copy because no change was noted in the organization of the reporting process between these two data collection rounds other than the introduction of the PACS. RESULTS: Data were collected on a total of 5568 report observations. Report time in the PACS data collection period was not significantly different (p = .32) than that in the computerized radiography hard-copy period. Reporting with the PACS was associated with significantly more (p < .01) historical images (i.e., images of the same patient obtained in previous examinations) being viewed. CONCLUSION: Report time was not lengthened by the introduction of the PACS. The finding that more historical images were viewed when the PACS was in use indicates that the PACS brought about a positive change in reporting practice.

Bias↗
PubMed1997

A nested set of C-terminal deletions of the alpha subunit of Escherichia coli RNA polymerase define regions concerned with assembly, proteolysis, stabilization and transcriptional activation in vivo.

BACKGROUND: The alpha subunit of eubacterial RNA polymerase comprises an N-terminal assembly domain and a mobile C-terminal domain which provides an activation contact site for class I transcription activators. One particular C-terminal alpha mutant, rpoA341, impairs the response of Escherichia coli RNA polymerase to several activators, including MelR. RESULTS: The in vivo properties of a set of C-terminally truncated alpha variants were investigated. Derivatives of 230 amino acids or longer were assembled into functional RNA polymerase. However, derivatives greater than 271 residues in length were sensitised to proteolysis near K271. Deletion of only 13 C-terminal amino acids impaired the response to CRP at a class I promoter whereas the complete removal of the alpha C-terminal domain did not prevent complementation of MelR-dependent PmelAB activity in the rpoA341 mutant. CONCLUSIONS: Our results refine the C-terminal limit of the alpha assembly domain to between residues 221 and 230. The 13 extreme C-terminal amino acids are exposed in the holoenzyme and participate in the protection of an otherwise proteolytically sensitive bond near K271. Their presence is also essential for transcription activation at class I CRP-dependent promoters. The rpoA341-mediated substitution, K271E, does not define an activation contact site for MelR.

Bacterial Proteins↗
PubMed1997

Release of atrial natriuretic factor prohormone peptides 1-30, 31-67 and 99-126 from freshwater- and seawater-acclimated perfused trout (Oncorhynchus mykiss) hearts

Atrial natriuretic factor (ANF), a 28-amino-acid peptide hormone produced in the heart, circulates in both freshwater and seawater rainbow trout. In mammals, two other peptide hormones, proANF 1-30 and proANF 31-67, derived from the same 126-amino-acid prohormone as ANF (amino acids 99&shyp;126), circulate and have natriuretic and diuretic properties. It has never been determined whether these peptides circulate in fish. The present investigation was designed to determine (1) whether proANF 1-30 and/or proANF 31-67 circulate in perfused hearts from freshwater- and seawater-acclimated rainbow trout (Oncorhynchus mykiss) in situ, and (2) if they do, to determine whether increasing the filling pressure of the heart causes their release in trout as it does in mammals. High-performance gel-permeation chromatography of fish plasma revealed that both proANF 1-30 and 31-67 circulate in freshwater- and seawater-acclimated trout plasma at threefold higher concentrations than does ANF. The basal rates of release of ANF and proANF 1-30 and 31-67 were similar in both freshwater and seawater trout, with the rate of release of proANF 1-30 being 10 times higher and that of proANF 31-67 20 times higher than that of ANF. When the filling pressure was increased to the peak of the Starling curve (max), the rate of release of ANF and proANFs 1-30 and 31-67 increased fivefold for each peptide in the freshwater trout, while in seawater trout the rates of release increased six- to ninefold. We conclude that proANF 1-30 and 31-67, as well as ANF, circulate in both freshwater-and seawater-acclimated trout and do so at concentrations higher than that of ANF. Increasing the filling pressure to the trout heart was found to cause a similar increase in the release rates for each of these peptides, but the maximal increase was higher in the seawater-acclimated trout, apparently because they showed a larger increase in cardiac output.

Journal Article↗
PubMed1996

Location of essential sequence elements at the Escherichia coli melAB promoter.

The Escherichia coli melAB promoter has been cloned on a short DNA fragment and subjected to deletion mutagenesis, random mutagenesis and site-directed mutagenesis. In previous work we had shown that expression from the melAB promoter is triggered by melibiose and that this requires the MelR transcription activator. Melibiose-dependent expression is suppressed by deletions that remove both DNA-binding sites for MelR and by point mutations in the -10 hexamer, the -35 hexamer and the region just upstream of the -35 hexamer. The point mutations identify promoter elements that are essential for triggering the melAB promoter. The importance of these elements was confirmed by site-directed mutagenesis. The results show that the organization of the melAB promoter is fundamentally different from the organization of other bacterial promoters controlled by homologues of MelR.

Base Sequence↗
PubMed1996

Ovine lentivirus antibody detection in serum, colostrum and milk using a recombinant transmembrane protein ELISA.

An enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against ovine lentivirus (OLV) in serum, colostrum, and milk from naturally infected sheep. The assay used OLV recombinant transmembrane envelope protein (rTM) as a test antigen. Matched serum/colostrum and serum/milk samples were collected at 24h, 4 weeks (mid-lactation), and 8 weeks (weaning) post-lambing. Among 129 paired samples collected at 24 h post-lambing, there was overall test agreement (concordance) of 82.9% and a kappa value of 0.658 between serum and colostrum rTM ELISA results. Among 130 mid-lactation samples, the milk ELISA had 100% specificity and 64.9% sensitivity relative to the serum ELISA, there was concordance of 79.2%, and a kappa value of 0.602. At mid-lactation, the serum agar gel immunodiffusion test had a sensitivity of 0.390 and 0.560 relative to the serum and milk rTM ELISAs, respectively. Matched serum and milk rTM ELISA results at weaning were very similar to those at mid-lactation. Finally, increased occurrence and severity of subclinical mastitis at weaning was found in ELISA-seropositive compared with ELISA-seronegative ewes. Both subclinical mastitis and ewe OLV infection had a negative impact on lamb growth and weaning weights. Compared with blood, colostrum and milk are easier and less expensive to sample and store. These results suggest that rTM ELISA testing of colostrum and milk could be used to supplement serologic testing in OLV screening or eradication programs.

Animals↗
PubMed1996

Characterization of enzootic nasal tumor virus capsid antigen.

The RT-PCR was carried out on tumor tissue from sheep with enzootic nasal tumor (ENT), using primers designed from conserved amino acid regions of related type D retroviruses. A 591 bp PCR fragment, corresponding to 90% of the capsid antigen was cloned, sequenced and expressed in E. coli. Alignment with ovine pulmonary carcinoma (OPC) virus showed 93% nucleotide and 96% amino acid homology. No amplification occurred when DNA from ovine fetal cell line was used as template. The recombinant protein, highly expressed in prokaryotic system, reacted in immunoblot with mouse antiserum to Mason Pfizer monkey virus (MPMV) p27, as well as sera from OPC and ENT diseased animals. Preliminary application of this antigen in ELISA suggested its potential use to detect seropositive animals in infected flocks.

Animals↗
PubMed1995

Case study evaluation.

Case study evaluations, using one or more qualitative methods, have been used to investigate important practical and policy questions in health care. This paper describes the features of a well designed case study and gives examples showing how qualitative methods are used in evaluations of health services and health policy.

Health Resources↗
PubMed1995

Comparison of ovine lentivirus detection by conventional and recombinant serological methods.

Recombinant (r) transmembrane protein (TM), major capsid protein P25, and matrix protein P16 of ovine lentivirus (OLV) were used as solid phase antigens in enzyme-linked immunosorbent assays (ELISAs) for the detection of specific antibodies against OLV in sheep sera. Sensitivity, specificity, and agreement of these three recombinant assays were compared with each other and with two currently available conventional OLV serological assays, the agar gel immunodiffusion (AGID) test and a whole-virus (WV) ELISA. Field sera from a total of 412 Midwestern United States sheep were tested and compared by the five OLV detection methods, including visibly healthy sheep selected for public sale (Group A, n = 171), samples from a breeding flock of Finnsheep and Finn-cross ewes (Group B, n = 184) and moribund sheep with clinical signs associated with OLV (Group C, n = 57). The rTM ELISA was the most sensitive OLV detection assay, both overall and within each group. Sera from 48.1% (198/412) of field samples were rTM ELISA positive. By contrast, positive rates for the rP25, rP16, and WV ELISAs and AGID test were 34.2%, 32.3%, 36.9%, and 26.9%, respectively. The rTM ELISA reactivity was 36.8% for Group A sera, 50.0% for Group B sera, and 75.4% for Group C sera. Among the 21 Group C sheep possessing OLV lung lesions at necropsy, 20 (95.2%) were rTM ELISA positive. The greatest test agreement occurred between the rP25 and the rP16 ELISAs. The data suggest that the recombinant TM immunoassay is the most accurate and sensitive of the five methods evaluated for the detection of serum anti-OLV antibodies in sheep, both at the subclinical infection and overt clinical disease stages.

Aging↗
PubMed1995

Development and application of an antibody ELISA for the marker protein of ovine pulmonary carcinoma.

Ovine pulmonary carcinoma (OPC) is a contagious pulmonary neoplasia with a suspected retroviral etiology. The major core protein (P27) of the putative OPC virus cross-reacts with antibodies to P27 of the Mason-Pfizer monkey virus (MPMV), a type-D retrovirus. This serological reactivity serves as the only accepted biological marker for OPC. In order to make a useful reagent for the detection of the OPC marker for serodiagnosis and epidemiological studies, the MPMV-P27 coding region was cloned and expressed in Escherichia coli. Gel purified recombinant MPMV-P27 protein was used to develop an immunoassay. This recombinant enzyme-linked immunosorbent assay (ELISA) was then used to screen 223 sera from US sheep and 176 sera from Italian sheep. In this study, we found: (1) a high prevalence of infection with the putative OPC retrovirus in sheep with chronic pneumonia; (2) a subclinical infection with OPC virus may be more common in US sheep than indicated by the rare recorded occurrence of pulmonary carcinoma; (3) an apparent association between ovine lentivirus (OLV) and OPC infection.

Adenocarcinoma, Bronchiolo-Alveolar↗
PubMed1995

Issues in the evaluation of picture archiving and communication systems.

Picture archiving and communication systems (PACS) are an example of the application of computer technology in the medical field. PACS automates image handling in a hospital and has the potential to transform the way radiology is currently performed. This paper focuses on the evaluation of the PACS technology, and considers the claims that have been made for PACS, how these claims might be turned into questions to be addressed by evaluation and the appropriate methods for the evaluation of PACS. A distinction is drawn between evaluation questions for which the hospital is the appropriate focus and those for which the patient is the appropriate focus. The preferred research design is different for hospital focused PACS evaluation and patient-focused evaluation of small scale PACS systems. A contemporaneous experimental comparison within hospitals is the preferred design for the patient-focused evaluation of small scale PACS systems. The patient-focused evaluation of large scale systems and the hospital-focused evaluation of all PACS systems could feasibly be conducted as contemporaneous experimental comparisons between hospitals but the large research costs implied by such a design almost certainly mean that non-contemporaneous, non-experimental comparisons within hospitals are more realistic. The current situation for the PACS technology is that it has potential, but as yet unproven, benefits and a large capital cost. Thus, the primary purpose of funding additional PACS implementations must be to add to the currently small body of evaluation evidence.

Cost Savings↗