Assessment of an ultrasound bladder scanner in prostate radiotherapy: A validation study and analysis of bladder filling variability.

INTRODUCTION: During prostate radiotherapy treatment, it is important to ensure the position of the bladder and prostate is consistent between treatments. The aim of this study was to provide a quantitative basis for incorporating ultrasound bladder volume estimates into local practice for prostate radiotherapy. METHODS: Agreement between bladder volume estimates obtained using computed tomography (CT) and ultrasound was assessed. Analysis of bladder volumes between planning and treatment scans was used to quantify expected variations in bladder volume over the course of radiotherapy. Dose-volume statistics were estimated and compared to planned dose constraints to propose a target bladder volume and tolerance. RESULTS: Bladder volume measurements were obtained from 19 radiotherapy patients using ultrasound and CT. Ultrasound underestimated bladder volume compared to CT with a mean bias of -28 ± 30 ml. Pre-treatment (planning) bladder volumes varied from 71 to 383 ml with a mean of 200 ml. Treatment bladder volumes reduced by more than half in 9% of patients during the course of their treatment, potentially leading to a 30% increase in mean bladder dose. Patients with pre-treatment bladder volumes < 200 ml were most likely to exhibit differences in bladder volume, resulting in 'out of tolerance' increases in dose. CONCLUSIONS: A pragmatic individualised drinking protocol, aimed at achieving a minimum ultrasound bladder volume of 200 ml at planning CT, may be beneficial to reproducibility in radiotherapy treatment. Ultrasound measurements prior to treatment should ideally confirm that bladder volume is at least half the volume measured at planning.

Biosecurity-based interventions and strategies to reduce Campylobacter spp. on poultry farms.

The prevention and control of Campylobacter colonization of poultry flocks are important public health strategies for the control of human campylobacteriosis. A critical review of the literature on interventions to control Campylobacter in poultry on farms was undertaken using a systematic approach. Although the focus of the review was on aspects appropriate to the United Kingdom poultry industry, the research reviewed was gathered from worldwide literature. Multiple electronic databases were employed to search the literature, in any language, from 1980 to September 2008. A primary set of 4,316 references was identified and scanned, using specific agreed-upon criteria, to select relevant references related to biosecurity-based interventions. The final library comprised 173 references. Identification of the sources of Campylobacter in poultry flocks was required to inform the development of targeted interventions to disrupt transmission routes. The approach used generally involved risk factor-based surveys related to culture-positive or -negative flocks, usually combined with a structured questionnaire. In addition, some studies, either in combination or independently, undertook intervention trials. Many of these studies were compromised by poor design, sampling, and statistical analysis. The evidence for each potential source and route of transmission on the poultry farm was reviewed critically, and the options for intervention were considered. The review concluded that, in most instances, biosecurity on conventional broiler farms can be enhanced and this should contribute to the reduction of flock colonization. However, complementary, non-biosecurity-based approaches will also be required in the future to maximize the reduction of Campylobacter-positive flocks at the farm level.

Potential sources of Campylobacter infection on chicken farms: contamination and control of broiler-harvesting equipment, vehicles and personnel.

AIMS: To test the efficacy of enhanced biosecurity measures on poultry farms for reducing environmental contamination with Campylobacter during partial depopulation of broiler flocks prior to normal slaughter age. The study has also evaluated the risk of infection from live-bird transport crates that are routinely cleaned at the slaughterhouse, but may remain contaminated. METHODS AND RESULTS: On-farm sampling and Campylobacter isolation was undertaken to compare the prevalence of contamination on vehicles, equipment and catching personnel during farm visits that took place under normal or enhanced biosecurity. Campylobacters were found in almost all types of sample examined and enhanced biosecurity reduced the prevalence. However, the additional measures failed to prevent colonisation of the flocks. For transport crates, challenge trials involved exposure of broilers to commercially cleaned crates and genotyping of any campylobacters isolated. The birds were rapidly colonised with the same genotypes as those isolated from the cleaned crates. CONCLUSIONS: The enhanced biosecurity measures were insufficient to prevent flock colonisation, and the problem was exacerbated by inadequate cleaning of transport crates at the slaughterhouse. SIGNIFICANCE AND IMPACT OF THE STUDY: Current commercial practices in the United Kingdom facilitate the spread of campylobacters among broiler chicken flocks. Prevention of flock infection appears to require more stringent biosecurity than that studied here.

An assessment of the microbiological risks involved with egg washing under commercial conditions.

The potential benefits of washing eggs is offset by a historical perception in the European Union that wetted eggs are prone to spoilage and water loss. This study describes the effects of spray jet washing under various processing conditions to shell surface counts of Salmonella and the presence of bacteria in egg contents. Experiments used eggs that were contaminated with Salmonella Enteritidis PT4 or Salmonella Typhimurium DT104 before cuticle hardening. Washing of contaminated eggs under optimum conditions resulted in a more than 5-log reduction of Salmonella counts from the shell surface. Salmonella was not isolated from the yolk or albumen of any egg washed by the optimal protocol, suggesting that when properly controlled, egg washing did not cause Salmonella to enter the contents. However, contamination did arise if strict control was not maintained over the wash and rinse water temperatures. Both Salmonella Enteritidis and Salmonella Typhimurium were shown to enter the egg contents when water temperatures were lowered, indicating that strict temperature control must be maintained in order to prevent the ingress of Salmonella into egg contents. Other washing machine parameters that were investigated did not significantly affect Salmonella entry into the egg contents but influenced shell surface kill levels to varying degrees.

A decision theoretic approach to sample size determination in clinical trials.

In this paper, we discuss a Behavioral Bayes approach to the determination of sample size in phase III clinical trials for which the data are assumed to come from a normal distribution for which the mean and variance are both unknown. Software is described which minimizes the expected net cost as a function of the sample size, thereby establishing the optimal sample size. This methodology extends previous work by the assumption of unknown variance. Numerical examples show that the more general model can have a large effect on the optimal sample size, compared with a procedure which uses the known variance model with an estimate of the variance.

Complications in foot and ankle arthroscopy.

Arthroscopy of the foot and ankle has become an important diagnostic and therapeutic tool for the orthopaedic surgeon. A thorough knowledge of foot and ankle anatomy and intraarticular anatomy is critical to avoid complications in foot and ankle arthroscopy. Numerous complications can occur in foot and ankle arthroscopy, such as neurologic, tendon, and ligament injuries, wound complications, infections, and instrument breakage. The most common complication is neurologic injury. The overall complication rate is 9%. Most complications associated with foot and ankle arthroscopy are transient and tend to resolve within 6 months. The only complication that persisted at 10 years followup was a neurologic injury, specifically, numbness at the incision site. Because the difficulty of procedures has increased, so has the complication rate. Knowledge of the more common complications in foot and ankle arthroscopy and improved techniques and instruments may reduce the overall complication rate.

Prediction of biological activity for high-throughput screening using binary kernel discrimination.

High-throughput screening has made a significant impact on drug discovery, but there is an acknowledged need for quantitative methods to analyze screening results and predict the activity of further compounds. In this paper we introduce one such method, binary kernel discrimination, and investigate its performance on two datasets; the first is a set of 1650 monoamine oxidase inhibitors, and the second a set of 101 437 compounds from an in-house enzyme assay. We compare the performance of binary kernel discrimination with a simple procedure which we call "merged similarity search", and also with a feedforward neural network. Binary kernel discrimination is shown to perform robustly with varying quantities of training data and also in the presence of noisy data. We conclude by highlighting the importance of the judicious use of general pattern recognition techniques for compound selection.

The DNA-binding domain of the gene regulatory protein AreA extends beyond the minimal zinc-finger region conserved between GATA proteins.

The AreA protein of Aspergillus nidulans regulates the activity of over 100 genes involved in the utilisation of nitrogen, and has a limited region of homology with the vertebrate family of GATA proteins around a zinc finger (Zf) motif. A 66 amino acid (a.a.) residue fragment (Zf(66)) corresponding to the zinc finger, a 91 a.a fragment (Zf(91)) containing an additional 25 a.a. at the C-terminus, and a much larger 728 a.a. sequence (3'EX) corresponding to the 3'exon have been over-expressed as fusion proteins in E. coli and purified. The DNA-protein complexes formed by these proteins have been examined by gel retardation analysis. The 91 a.a. protein forms a discrete shifted species with a GATA-containing DNA fragment with high affinity (K(d)=0.15 nM), whereas the 66 a.a. protein has very low ( approximately microM) affinity for the same sequence. The results show that the region of AreA required for high affinity DNA binding extends beyond the zinc finger motif that is homologous to GATA-1, requiring in addition a region within the 25 a.a. sequence C-terminal to the zinc finger. Using hydroxyl radical and ethylation interference footprinting, the minimal Zinc finger protein (Zf(66)) shows no appreciable interference effects whereas Zf(91) shows much stronger interference effects, identical to those of the larger protein. These effects extend over sequences up to two nucleotides either side of the GATA site, and indicate contacts additional to those observed in the three-dimensional structure of the complex of the minimal zinc-finger protein with DNA. We suggest that these additional contacts are responsible for the enhanced DNA binding affinity of the extended zinc-finger protein Zf(91).

Transgene expression driven by heterologous ribulose-1, 5-bisphosphate carboxylase/oxygenase small-subunit gene promoters in the vegetative tissues of apple (Malus pumila mill.).

It is desirable that the expression of transgenes in genetically modified crops is restricted to the tissues requiring the encoded activity. To this end, we have studied the ability of the heterologous ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) small-subunit (SSU) gene promoters, RBCS3CP (0.8 kbp) from tomato (hycopersion esculentum Mill.) and SRS1P (1.5 kbp) from soybean (Glycine max [h.] Mers.), to drive expression of the beta-glucuronidase (gusA) marker gene in apple (Malus pumila Mill.). Transgenic lines of cultivar Greensleeves were produced by Agrobacterium-mediated transformation and the level of gusA expression in the vegetative tissues of young plants was compared with that produced using the cauliflower mosaic virus (CaMV) 35S promoter. These quantitative GUS data were assessed for their relationship to the copy number of transgene loci. The precise location of GUS activity in leaves was identified histochemically. The heterologous SSU promoters were active primarily in the green vegetative tissues of apple, although activity in the roots was noticeably higher with the RBCS3C promoter than with the SRS1 promoter. The mean GUS activity in leaf tissue of the SSU promoter transgenics was approximately half that of plants containing the CaMV 35S promoter. Histochemical analysis demonstrated that GUS activity was localised to the mesophyll and palisade cells of the leaf. The influence of light on expression was also determined. The activity of the SRS1 promoter was strictly dependent on light, whereas that of the RBCS3C promoter appeared not to be. Both SSU promoters would be suitable for the expression of transgenes in green photosynthetic tissues of apple.

Electrical stimulation as a therapeutic option to improve eyelid function in chronic facial nerve disorders.

PURPOSE: To establish whether it is possible to improve orbicularis oculi muscle function in the eyelids of patients with a chronic seventh cranial nerve palsy by using transcutaneous electrical stimulation to the point at which electrical stimulation induces a functional blink. METHODS: Ten subjects (one woman, nine men) aged 36 to 76 with chronic, moderate to severe facial nerve palsy were recruited into the study. Voluntary and spontaneous eyelid movements were assessed, using an optical measuring system, before, during, and after a 3-month treatment period. Voluntary and spontaneous lid velocities were also measured and compared with eyelid kinematic data in normal subjects (12 women, 18 men; age range, 22-56 years). RESULTS: Therapeutic electrical stimulation applied over 3 months produced improvement in eyelid movement (>2 mm) in 8 of 10 patients during voluntary eyelid closure. However, there was no significant improvement recorded in spontaneous blink amplitudes or peak downward-phase velocity of the upper eyelid. This regimen of stimulation failed to recover function well enough that a functional blink could be induced in the paretic eyelid by electrical stimulation. CONCLUSIONS: Electrical stimulation using transcutaneous electrical nerve stimulators units can improve voluntary eye closure, apparently because of a reduction in stiffness of eyelid mechanics, rather than an improvement of muscle function. Investigation of alternative stimulation regimens is warranted.

Performance characteristics of the Pantak DXT-300 kilovoltage X-ray treatment machine.

The beam and performance characteristics of a new orthovoltage X-ray unit, the Pantak DXT-300 have been evaluated. Data were collated for four qualities: 3.27 mmAl, 7.15 mmAl, 1.65 mmCu and 3.51 mmCu half value layer (HVL) (SE = 0.04 mm). Parameters which were investigated included beam quality, central axis depth dose, relative output, backscatter factors, field uniformity, peripheral dose and head leakage. The calibration procedure and the performance of the dosimetry system have also been described.

A line imaging system for measurement of eyelid movements.

A line scan imaging system which provides a facility for comprehensive measurement and analysis of eyelid motion in human subjects is described. The device obtains parasagittal line images of the eye and eyelids by focusing an image of the eye onto a linear 256-element CCD array. Line images are acquired and digitized at a rate of 200 per second and stored in a buffer memory. The stored data are subsequently analysed on a PC to give measurements of eyelid displacement and velocity. Measurements of eyelid displacement and velocity during normal blinks in five subjects were comparable with published results from other measurement techniques.

Multiple mechanisms of membrane anchoring of Escherichia coli penicillin-binding proteins.

The major penicillin-binding proteins (PBPs) of Escherichia coli play vital roles in cell wall biosynthesis and are located in the inner membrane. The high M(r) PBPs 1A, 1B, 2 and 3 are essential bifunctional transglycosylases/transpeptidases which are thought to be type II integral inner membrane proteins with their C-terminal enzymatic domains projecting into the periplasm. The low M(r) PBP4 is a DD-carboxypeptidase/endopeptidase, whereas PBPs 5 and 6 are DD-carboxypeptidases. All three low M(r) PBPs act in the modification of peptidoglycan to allow expansion of the sacculus and are thought to be periplasmic proteins attached with varying affinities to the inner membrane via C-terminal amphiphilic alpha-helices. It is possible that the PBPs and other inner membrane proteins form a peptidoglycan synthesizing complex to coordinate their activities.