Comparing real-time and conventional PCR to culture-based methods for detecting and quantifying Escherichia coli O157 in cattle feces.

Detection of Escherichia coli O157 in cattle feces has traditionally used culture-based methods; PCR-based methods have been suggested as an alternative. We aimed to determine if multiplex real-time (mq) or conventional PCR methods could reliably detect cattle naturally shedding high (≥10(4) CFU/g of feces) and low (∼10(2) CFU/g of feces) concentrations of E. coli O157. Feces were collected from pens of feedlot cattle and evaluated for E. coli O157 by culture methods. Samples were categorized as (i) high shedders, (ii) immunomagnetic separation (IMS) positive after enrichment, or (iii) culture negative. DNA was extracted pre- and postenrichment from 100 fecal samples from each category (high shedder, IMS positive, culture negative) and subjected to mqPCR and conventional PCR assays based on detecting three genes, rfbE, stx1, and stx2. In feces from cattle determined to be E. coli O157 high shedders by culture, 37% were positive by mqPCR prior to enrichment; 85% of samples were positive after enrichment. In IMS-positive samples, 4% were positive by mqPCR prior to enrichment, while 43% were positive after enrichment. In culture-negative feces, 7% were positive by mqPCR prior to enrichment, and 40% were positive after enrichment. The proportion of high shedder-positive and culture-positive (high shedder and IMS) samples were significantly different from mqPCR-positive samples before and after enrichment (P < 0.01). Similar results were observed for conventional PCR. Our data suggest that mqPCR and conventional PCR are most useful in identifying high shedder animals and may not be an appropriate substitute to culture-based methods for detection of E. coli O157 in cattle feces.

Evidence of non-O157 Shiga toxin-producing Escherichia coli in the feces of meat goats at a U.S. slaughter plant.

Shiga toxin-producing Escherichia coli (STEC) are important human pathogens, and attention to non-O157 serogroups has increased in recent years. Although cattle are normally considered the primary reservoir for STEC, recent illnesses associated with goat contact have indicated that these animals are important potential reservoirs for the organisms. The prevalence of STEC, particularly non-O157 serogroups, in U.S. goats has not been well described. Our objective was to determine the prevalence of six major non-O157 STEC serogroups in the feces of meat goats. Rectal contents from 296 goats were collected postevisceration at a slaughter plant in the southeastern United States over 9 days during a 12-week period from August through October 2012. Samples were enriched in E. coli broth, and DNA was extracted and used as template in an 11-gene multiplex PCR that detected six non-O157 serogroups (O26, O45, O103, O121, O111, and O145) and virulence genes. Samples were considered positive when at least one non-O157 STEC serotype was present with either stx1 or stx2. All six non-O157 serogroups were detected by PCR in our samples, and 14.5% of samples were positive for at least one serogroup. Prevalence of O26 was highest, with 6.4% of goat fecal samples positive. The prevalence of O45 was 3.4%, O103 was 4.4%, O111 was 4.1%, O121 was 1.4%, and O145 was 3.0%. Twenty-two (7.4%) of 296 fecal samples had more than one non-O157 serogroup detected in the feces. Two samples had evidence of three non-O157 STEC serogroups. Goats appear to be an important reservoir for non-O157 STEC, and further work to understand the characteristics, epidemiology, and ecology of STEC in these animals is warranted.

Prevalence and relatedness of Escherichia coli O157:H7 strains in the feces and on the hides and carcasses of U.S. meat goats at slaughter.

We determined the prevalences of Escherichia coli O157:H7 in feces, hide, and carcasses of meat goats at a U.S. processing plant. Prevalences were 11.1%, 2.7%, and 2.7%, respectively. Sixteen pulsed-field gel electrophoresis (PFGE) subtypes were identified among 49 E. coli O157:H7 isolates, some of which were present on multiple sample types or collection days.

A mathematical model for patient skin dose assessment in cardiac catheterization procedures.

A mathematical model has been developed for the assessment of patient skin doses from cardiac catheterization procedures. This uses exposure and projection data stored in the DICOM image files. Since these contain only information about the acquisition runs, a correction is needed to estimate and include the contribution from fluoroscopy. Maximum skin doses calculated by the model were found to correlate well with those measured on Kodak EDR2 film. Three methods for including the contribution from fluoroscopy were investigated, and all successfully identified patients receiving skin doses in excess of 1 Gy. It is hoped to automate this tool for routine assessment of skin doses in our cardiac catheterization laboratories.

Kodak EDR2 film for patient skin dose assessment in cardiac catheterization procedures.

Patient skin doses were measured using Kodak EDR2 film for 20 coronary angiography (CA) and 32 percutaneous transluminal coronary angioplasty (PTCA) procedures. For CA, all skin doses were well below 1 Gy. However, 23% of PTCA patients received skin doses of 1 Gy or more. Dose-area product (DAP) was also recorded and was found to be an inadequate indicator of maximum skin dose. Practical compliance with ICRP recommendations requires a robust method for skin dosimetry that is more accurate than DAP and is applicable over a wider dose range than EDR2 film.

Film dosimetry for fluoroscopic procedures: potential errors.

Kodak EDR 2 film was calibrated across the range of beam qualities and exposure rates typically used in our cardiac catheterisation laboratory. Its dose-response curve was successfully modelled up to its saturation point of 1 Gy. The consistency of the film's response with film batch, time between exposure and processing processor and day-to-day variations in performance, was investigated. The effects of field size, exposure rate, beam quality and use of the dynamic wedge filter were quantified. The overall uncertainty in dose was estimated to be between -20% and +40%, at 160 mGy. This uncertainty increases as the film approaches its saturation point. In addition, some unusual artefacts were observed.

Glutamate-gated chloride channels and the mode of action of the avermectin/milbemycin anthelmintics.

The macrocyclic lactones are the biggest selling and arguably most effective anthelmintics currently available. They are good substrates for the P-glycoproteins, which might explain their selective toxicity for parasites over their vertebrate hosts. Changes in the expression of these pumps have been implicated in resistance to the macrocyclic lactones, but it is clear that they exert their anthelmintic effects by binding to glutamate-gated chloride channels expressed on nematode neurones and pharyngeal muscle cells. This effect is quite distinct from the channel opening induced by glutamate, the endogenous transmitter acting at these receptors, which produces rapidly opening and desensitising channels. Ivermectin-activated channels open very slowly but essentially irreversibly, leading to a very long-lasting hyperpolarisation or depolarisation of the neurone or muscle cell and therefore blocking further function. Molecular and genetic studies have shown that there are multiple GluCl isoforms in both free-living and parasitic nematodes: the exact genetic make-up and functions of the GluCl may vary between species. The known expression patterns of the GluCl explain most of the observed biological effects of treatment with the macrocyclic lactones, though the reason for the long-lasting inhibition of larval production in filarial species is still poorly understood.

Barium enema: use of increased copper filtration to optimize dose and image quality.

The purpose of this study was to determine and validate the optimum copper filtration for adult double contrast barium enema examinations. Entrance surface dose rates to polymethyl methacrylate slabs and corresponding image intensifier input kermas, were measured for various added copper filters. Image contrast was assessed using a Leeds TO.10 test object. Copper filter thickness of 0.3 mm was chosen, as this reduced entrance surface dose rate by 56%, without substantially degrading image contrast due to kV and mA saturation. 20 sets of clinical films taken with each of 0.3 mm copper, 0.1 mm copper and no copper were reviewed following randomization, by a specialist gastrointestinal radiologist. Each set of digital spot and conventional films was allocated a score for each of three regions of the bowel, on a scale of 0-3 for perceived barium coating. The Kruskal-Wallis test showed no significant difference in perceived coating between the three groups (Digital spot: sigmoid colon p=0.207, splenic flexure p=0.103, hepatic flexure p=0.894. Screen-film: left colon p=0.803, right colon p=0.487, transverse colon p=0.905). All examinations but one were classified as diagnostic. The remaining one was classified indeterminate, due to poor distension of the colon. On adding 0.3 mm copper filtration, the mean dose-area product per examination was reduced by 57%, from 17.7 Gy cm(2) to 7.6 Gy cm(2). The estimated reduction in effective dose was 11%, from 3.0 mSv to 2.7 mSv. X-ray tube loading increased by 30%, but this caused no overheating with our local examination protocol and schedule. Additional filtration of 0.3 mm copper for adult double contrast barium enemas has now been implemented in routine clinical use at our hospital.

Development of a neuronal pressure sensor.

Neuronal sensory systems are capable of performing very complex signal processing functions. Reconstruction of such sensory systems in vitro should enable whole-cell biological sensors to be generated that possess inherent signal processing capabilities. In this paper, the results of preliminary investigations to produce a mechanosensory neuronal network are presented. An in vitro network of rat dorsal root ganglion neurons has been produced on a microelectrode plate revealing an interesting rhythmical pattern of spontaneous discharges. This periodic activity has been shown to be disrupted following the application of a static pressure to the cell culture. These results indicate that neuronal networks represent a practical system that may be used for the development of intelligent, whole-cell, biological sensors.

A generic approach to risk assessment for the Ionising Radiations Regulations 1999.

The Ionising Radiations Regulations 1999 (IRR99) require that an explicit risk assessment be carried out to control properly the risks associated with the use of ionising radiation. Prior to IRR99, risk was controlled implicitly via mechanisms such as Local Rules and staff monitoring, which were initiated by the Radiation Protection Adviser using professional judgement. To enable a consistent approach across our Trust's radiation users, a generic model of risk assessment was employed. This had been developed previously by the Trust in response to a national initiative within the NHS to manage risk. The methodology involves the generation, by a multidisciplinary staff brain-storming session, of a complete list of all possible risks. This is followed by a risk stratification process based upon severity of consequence and frequency of risk. Auditable controls are then applied and quantifiable outcome measures are monitored for continued compliance. Template risk assessments are presented to aid others in this process. It is estimated that even in a large teaching hospital with a full complement of medical radiation applications, the time resource necessary for this approach is of the order of a few days.

Dose-area product measurements in a range of common orthopaedic procedures and their possible use in establishing local diagnostic reference levels.

There is a national drive towards establishing reference doses for radiological procedures with the aim of optimizing patient doses. Furthermore, the establishment of diagnostic reference level doses became a legal requirement for all hospitals on 13 May 2000. However, to date there are little published data on patient radiation doses from fluoroscopic procedures during orthopaedic surgery. Data relating to patient dose for 492 patients undergoing fluoroscopic examinations during a range of surgical orthopaedic procedures in 1997-1998 have been analysed. Median dose-area product (DAP) readings and interquartile ranges for a variety of common fluoroscopic orthopaedic procedures are presented. In general, the median DAP for procedures on limbs and extremities was quite low (0.04-1.62 Gy cm2), with screening times in the range 0.2-2.0 min, whilst for procedures involving the hips and spine the median DAP was considerably higher (0.4-10.2 Gy cm2), although overall screening times were similar, in the range 0.2-1.4 min. Approximate effective doses have been estimated. For procedures involving the limbs and extremities, the effective dose was generally less than 10 microSv, and for procedures involving the hips and spine it was found that the effective dose could rise to about 1 mSv. Collective doses for each procedure have been calculated to inform prioritization of procedures for local dose reduction strategies.

Prophylactic nitroglycerin did not reduce myocardial ischemia during accelerated recovery management of coronary artery bypass graft surgery patients.

OBJECTIVE: To evaluate the use of a high dose of nitroglycerin (NTG) for prophylaxis against myocardial ischemia and infarction in patients undergoing coronary artery bypass graft (CABG) surgery with accelerated recovery. DESIGN: Prospective, double-blind, placebo-controlled randomized study. SETTING: A university-based medical center. PARTICIPANTS: Forty adult patients presenting for elective CABG surgery. INTERVENTIONS: Forty patients were divided into 2 blinded study groups. Twenty patients received 2 microg/kg/min of NTG starting before induction of anesthesia and continuing for 6 hours after extubation in the intensive care unit. The placebo group (n = 20) received normal saline during this same interval. MEASUREMENTS AND MAIN RESULTS: Hemodynamics, incidence and severity of myocardial ischemia, and myocardial infarction rates were determined. There were no differences in hemodynamic parameters between groups. The incidence of ischemia was approximately 35% in each group. Myocardial infarction (as determined by elevated creatine kinase-MB fraction, troponin I, and electrocardiogram criteria) was 10% in the placebo group and 5% in the NTG group (p = 0.234). CONCLUSIONS: This study shows a high incidence of myocardial ischemia and infarction in patients presenting for CABG surgery with an accelerated recovery management scheme. NTG was well tolerated clinically; however, it was not found to be protective against myocardial ischemia or infarction in this setting.

Age and other risk factors for neuropsychologic decline in patients undergoing coronary artery bypass graft surgery.

OBJECTIVE: To evaluate if increasing age is a major determinant of long-term neuropsychologic (NP) dysfunction in patients undergoing coronary artery bypass graft (CABG) surgery. DESIGN: Prospective cohort study. SETTING: A single university hospital. PARTICIPANTS: Three hundred eighty-one patients undergoing elective CABG surgery. INTERVENTIONS: Baseline preoperative and intraoperative characteristics were assessed for all patients. A comprehensive NP test battery was conducted preoperatively and at 1 month postoperatively. MEASUREMENTS AND MAIN RESULTS: Of the initial 381 patients, 319 (84%) returned for the 1-month visit. The overall incidence of an NP deficit was 21.6%. There was no significant difference seen with increasing age, although a trend was observed with highest NP deficit rate in the older than 65 age group. Other preoperative risk factors, including gender, education level, and preexisting diseases, were nonsignificant. Intraoperative variables were evaluated and revealed no significant differences across the age groups. CONCLUSIONS: This cohort study did not find an increased incidence of NP deficits in elderly patients after CABG surgery. It has addressed the limitations of other studies, however, with (1) a comprehensive NP test battery, (2) longer-term follow-up, and (3) adequate sample size.

Cross-calibration, precision and patient dose measurements in preparation for clinical trials using dual energy X-ray absorptiometry of the lumbar spine.

Large, multicentre clinical trials using DXA to monitor bone density following intervention are now common. At the same time, several different bone densitometers and calibration phantoms are currently in use. The aim of this study was to document the technical information required on cross-calibration of equipment, reproducibility and patient dose before commencing a multicentre clinical trial. To this end, we obtained an in vitro and in vivo cross-calibration of two machines (a Hologic QDR 2000 and a Lunar DPX-L) that were not significantly different. Interobserver and intraobserver precision, and radiation dose were also measured and three commonly used phantoms assessed for their usefulness in cross-calibration and quality assurance. Measured in vitro precision on the two machines (0.3-0.7%) was better than that specified by the manufacturers. In vivo precision was worse (1.4-2.1%), as might be expected in patients with reduced bone mass. Mean entrance skin radiation doses on each machine were 280 microSv for the QDR 2000 and 38 microSv for the DPX-L. No one phantom is ideal, but the European Spine Phantom or Lunar Aluminium Spine Phantom will provide an adequate cross-calibration for a clinical trial. This study demonstrates that an adequate cross-calibration can be obtained for use in groups of patients and that the equipment used is reproducible with a low radiation output.

Risk factors and solutions for the development of neurobehavioral changes after coronary artery bypass grafting.

BACKGROUND: As operative mortality for coronary artery bypass grafting has decreased, greater attention has focused on neurobehavioral complications of coronary artery bypass grafting and cardiopulmonary bypass. METHODS: To assess risk factors and to evaluate changes in surgical technique, between 1991 and 1994 we evaluated 395 patients undergoing coronary artery bypass grafting with an 11-part neurobehavioral battery administered preoperatively and at 1 and 6 weeks postoperatively. Patients were instrumented with 5-MHz focused continuous-wave carotid Doppler transducers intraoperatively to estimate cerebral microembolism as an instantaneous perturbation of the velocity signal. Microembolism data were quantitated and compared with surgical technical maneuvers during operation and with neurobehavioral deficit (> or = 20% decline from preoperative performance on two or more neurobehavioral tests) postoperatively. These data and patient demographics were statistically analyzed (chi2, t test) and the results at 2 years (1991 and 1992; group A) were used to influence surgical technique in 1993 and 1994 (group B). RESULTS: Significantly associated with new neurobehavioral deficits were increasing patient age (p < 0.05), more than 100 emboli per case (p < 0.04), and palpable aortic plaque (p < 0.02). Group B patients had a significant decline in the neurobehavioral event rate (group A, 69%, 140/203; versus group B, 60%, 115/192; p < 0.05) of postoperative neurobehavioral deficits at 1 week and at 1 month (group A, 29%, 52/180; versus group B, 18%, 35/198; p < 0.01). The stroke rate was less than 2% in both groups (p = not significant). Modifications of surgical technique used in group B patients included increased use of single cross-clamp technique, increased venting of the left ventricle, and application of transesophageal and epiaortic ultrasound scanning to locate and avoid trauma to aortic atherosclerotic plaques. CONCLUSIONS: Neurobehavioral changes after coronary artery bypass grafting are common and associated with cerebral microembolization. Surgical technical maneuvers designed to reduce emboli production may improve neurobehavioral outcome.

External radiation protection audit in National Health Service hospitals. Radiation protecgtion advisors (RPA) of the Nottingham City Hospital NHS Trust and the Oxford Radcliffe Hospital NHS Trust.

Radiation protection advisers are accustomed to carrying out internal audit in organizations for which they provide services but, traditionally, external audit has not been a part of the radiation protection adviser function. To provide an external audit function, two teams of radiation protection advisers undertook external radiation protection audits of their respective organizations, including the radiation protection adviser function itself. These audits were more representative of audits or inspections carried out by government agencies, for example the Health and Safety Executive and the former Inspectorate of Pollution, now the Environment Agency. Subsequently, reports, including recommendations for remedial action, were prepared for each management by the external auditors. The process of setting up and carrying out the external audits is described. Guidelines are suggested for those seeking to carry out an external audit.