Factors affecting the species of Campylobacter colonizing chickens reared for meat.

AIM: To investigate factors influencing Campylobacter spp. colonization of broiler chickens. METHODS AND RESULTS: Campylobacters were isolated from caeca from 319 flocks of two different breeds (199 Cobb and 120 Hubbard), reared as standard (199), Freedom Food/corn fed (57), free-range (47) or organic (16). The standard category exclusively used Cobb birds slaughtered at 38-41 days. The Freedom Food/corn-fed and free-range Hubbard birds were slaughtered at 49-56 days and the organic flocks at 70 days. Campylobacters were picked at random from direct plates. Both breed of chicken (Hubbard) and age at slaughter were independently associated with increased likelihood of colonization by Campylobacter coli rather than Campylobacter jejuni, but breed could not be separated from other aspects of husbandry with the data available. CONCLUSIONS: Chickens are frequently colonized by C. jejuni and C. coli and most human infections originate from poultry. In most developed countries approximately 90% of human infections are caused by C. jejuni, but fewer than 10% by C. coli. This might be due to C. coli being less pathogenic than C. jejuni to humans, and/or to chicken meat carrying fewer C. coli than C. jejuni. More investigations are needed into these aspects before it can be concluded that slaughtering older birds from slower-growing breeds would reduce the risk of human Campylobacter disease. SIGNIFICANCE AND IMPACT OF THE STUDY: Meat from certain breeds of poultry are predominantly colonized by C. coli rather than C. jejuni. More research is needed to understand the impact this may have on the number and severity of human campylobacter infections.

Involvement of homocysteine, homocysteine thiolactone, and paraoxonase type 1 (PON-1) in the etiology of defective human sperm function.

This study reports, for the first time, the significant (p ≤ 0.01) accumulation of homocysteine residues in low density, defective sperm suspensions isolated from patients attending an infertility clinic. This overabundance of homocysteine was not related to a deficiency in folate availability but may have been a reflection of the oxidative stress that characterizes such defective sperm populations. Direct addition of the homocysteine cyclic congener, homocysteine thiolactone, to human spermatozoa resulted in the rapid induction of mitochondrial reactive oxygen species (ROS) generation (p < 0.001), the stimulation of lipid peroxidation (p < 0.01), the promotion of tyrosine phosphorylation (p < 0.001), and the suppression of sperm motility (p < 0.001) in the absence of any significant impact on DNA integrity. The parent homocysteine molecule was less active and took 24 h to stimulate mitochondrial ROS production possibly because of the need to convert this compound to the corresponding thiolactone before it could exert a measureable biological effect. Thiolactone was also effective in suppressing the carboxymethylation of key proteins in the sperm tail, which are thought to be involved in the regulation of sperm movement. The major enzyme responsible for removing thiolactone from proteins, paraoxonase (PON-1), was shown to be a major target for alkylation by lipid aldehydes, such as 4-hydroxynonenal, generated as a consequence of oxidative stress. Exposure of human spermatozoa to such aldehydes resulted in a dose-dependent accumulation of homocysteine in spermatozoa (p < 0.03). These results suggest that one of the consequences of oxidative stress in mammalian spermatozoa is the inhibition of PON-1, which then enhances the availability of homocysteine thiolactone to interact with the epsilon-amino group of lysine residues on sperm proteins, triggering a raft of significant biological changes in these cells that ultimately compromise sperm function.

Pharmacokinetics of buprenorphine following intravenous and buccal administration in cats, and effects on thermal threshold.

This study reports the pharmacokinetics of buprenorphine, following i.v. and buccal administration, and the relationship between buprenorphine concentration and its effect on thermal threshold. Buprenorphine (20 µg/kg) was administered intravenously or buccally to six cats. Thermal threshold was determined, and arterial blood sampled prior to, and at various times up to 24 h following drug administration. Plasma buprenorphine concentration was determined using liquid chromatography/mass spectrometry. Compartment models were fitted to the time-concentration data. Pharmacokinetic/pharmacodynamic models were fitted to the concentration-thermal threshold data. Thermal threshold was significantly higher than baseline 44 min after buccal administration, and 7, 24, and 104 min after i.v. administration. A two- and three-compartment model best fitted the data following buccal and i.v. administration, respectively. Following i.v. administration, mean ± SD volume of distribution at steady-state (L/kg), clearance (mL·min/kg), and terminal half-life (h) were 11.6 ± 8.5, 23.8 ± 3.5, and 9.8 ± 3.5. Following buccal administration, absorption half-life was 23.7 ± 9.1 min, and terminal half-life was 8.9 ± 4.9 h. An effect-compartment model with a simple effect maximum model best predicted the time-course of the effect of buprenorphine on thermal threshold. Median (range) ke0 and EC50 were 0.003 (0.002-0.018)/min and 0.599 (0.073-1.628) ng/mL (i.v.), and 0.017 (0.002-0.023)/min and 0.429 (0.144-0.556) ng/mL (buccal).

Impact of the blood sampling site on time-concentration drug profiles following intravenous or buccal drug administration.

The aim of this study was to examine the effect of the sampling site on the drug concentration-time profile, following intravenous or buccal (often called 'oral transmucosal') drug administration. Buprenorphine (20 µg/kg) was administered IV or buccally to six cats. Blood samples were collected from the carotid artery and the jugular and medial saphenous veins for 24 h following buprenorphine administration. Buprenorphine concentration-time data were examined using noncompartmental analysis. Pharmacokinetic parameters were compared using the Wilcoxon signed rank test, applying the Bonferroni correction. Significance was set at P < 0.05. Following IV administration, no difference among the sampling sites was found. Following buccal administration, maximum concentration [jugular: 6.3 (2.9-9.8), carotid: 3.4 (1.9-4.9), medial saphenous: 2.5 (1.7-4.1) ng/mL], area under the curve [jugular: 395 (335-747), carotid: 278 (214-693), medial saphenous: 255 (188-608) ng·min/mL], and bioavailability [jugular: 47 (34-67), carotid: 32 (20-52), medial saphenous: 23 (16-55)%] were higher in the jugular vein than in the carotid artery and medial saphenous vein. Jugular venous blood sampling is not an acceptable substitute for arterial blood sampling following buccal drug administration.

The National Stroke Strategy - is it achievable?

INTRODUCTION: Timing of intervention in symptomatic carotid disease is critical. The UK Department of Health's National Stroke Strategy published in December 2007 recommends urgent carotid intervention within 48 h, in appropriate patients, who have suffered a transient ischaemic attack (TIA), amaurosis fugax or minor stroke. Despite the running of a rapid-access clinic for patients with symptoms of TIA, the time from symptom to surgery is rarely less than 2 weeks. To date, there has been little published research on the UK public response to the symptoms of TIA, and no study at all of the response of primary care to such patients. The aim of this study was to ascertain both these responses to see whether a 48-h target is achievable. PATIENTS AND METHODS: A total of 402 men attending our aortic aneurysm screening sessions were asked to complete a questionnaire requesting their most likely response to an episode of amaurosis fugax or TIA. All 45 GP practices in the hospital catchment area were asked how they would respond to patients requesting to be seen with the symptoms used in the questionnaire. RESULTS: Nearly one in six patients would ignore the symptom unless it recurred, approximately half would request a GP appointment and a third would see an optician if they had amaurosis fugax. The mean waiting time to see a GP was 2 days for a routine appointment and within 24 h for an emergency appointment. CONCLUSIONS: It is clear that a significant number of people would ignore the first symptom of carotid ischaemia; for those with amaurosis fugax, nearly a third would initially seek help from their optician. Those given a routine GP appointment would have to wait a minimum of 2 days. If the Department of Health is serious about reducing the incidence of stroke and introducing a target of 48 h from symptom to treatment, then there needs to be a wide-spread public and healthcare education programme, in particular alerting opticians and GP receptionists that these symptoms constitute a medical emergency.

A method to apply the robust estimator of dispersion, Qn, to fully-nested designs in the analysis of variance of microbiological count data.

This note outlines a method for the application of Rousseeuw's robust estimator of scale, Q(n), to fully-nested designs such as those used to estimate the components of variance in collaborative trials employed in quantitative food microbiology. A computer program is freely available on-line.

Estimates of measurement uncertainty from proficiency testing schemes, internal laboratory quality monitoring and during routine enforcement examination of foods.

AIMS: To determine the levels of measurement uncertainty (MU) obtained in proficiency testing and routine microbiological analyses of foods and to compare these with estimates of MU obtained for results of analyses obtained in collaborative interlaboratory studies of microbiological methods. METHODS AND RESULTS: Raw data submitted by participants in the Food Examination Proficiency Assessment Scheme were obtained from the Central Science Laboratory (York). Internal quality monitoring data were obtained from Health Protection Agency (HPA) laboratories, together with data from routine food examinations undertaken in HPA laboratories. The data sets were analysed to determine the relative standard deviations of reproducibility (RSD(R)), based on log(10) colony count values, and thence the relative measures of expanded uncertainty. Analysis of proficiency test data showed extreme values of RSD(R) up to +/-30% depending upon the organism, the laboratory and the method of examination. RSD(R) values on routine samples averaged around +/-12% but ranged up to +/-41% in a few instances. Internal quality assessments for different organisms ranged up to +/-27%, depending upon the particular organism and examination procedure. The results show little difference in uncertainty for counts obtained using different plating systems (e.g. pour plates, spread plates or spiral plating) on the same dilutions of the same food samples. The data are compared with estimates of microbiological uncertainty derived in interlaboratory studies. CONCLUSIONS: The estimates of uncertainty ranged widely, both within and between laboratories, and appeared to bear little relationship to the foodstuff under examination. The extent of MU associated with many routine microbiological examinations is generally no worse than those produced in inter-laboratory trials, although notable exceptions were seen. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of the levels of MU may have wide impact on the establishment of international standard methods for microbiological examination of foods and the ability to set realistic microbiological criteria.

Influence of presentation on outcome in abdominal aortic aneurysm repair.

OBJECTIVES: In the absence of formal screening abdominal aortic aneurysms (AAA) are detected in an opportunistic manner. Many remain asymptomatic and undetected until they rupture. Incidentally discovered small AAAs are entered into a surveillance programme until they reach a suitable size for repair. The aim of this study was to examine trends in the management of AAA and whether the method of presentation had an effect on subsequent mortality. DESIGN: Observational study in UK district general hospital. MATERIALS/METHODS: This study reports a single surgeon case series identified using a prospectively maintained database. Data on mode of presentation, management and mortality were retrieved from case notes, PIMS hospital database and the Office of National Statistics. RESULTS: Two hundred and five patients were referred with AAAs between 1992 and 2004, 78% presenting in elective circumstances. The surveillance programme fed 33% of the operated cases. Two aneurysms ruptured whilst under surveillance. Overall elective operative mortality was 11.8% and has progressively decreased over time. Thirty-day operated mortality was significantly lower in patients having a period of surveillance than those having immediate elective repair (2.3 vs. 16.3%, p=0.018). A slight reduction in emergency AAA repairs was noted over the study period (r2=0.6) although registered aneurysm deaths continue to increase (r2=0.83). CONCLUSIONS: Elective mortality following AAA surgery decreased over the study period. Outcome was better in those patients who had surgery for aneurysms that had been under surveillance. Despite opportunistic screening the population adjusted mortality rate of aortic aneurysms showed a progressive increase. A reduction in deaths from aneurysms is unlikely without a formal screening programme.

Estimating the precision of serial dilutions and colony counts: contribution of laboratory re-calibration of pipettes.

The basic (inherent) precision of serial dilutions and of colony counts made from them may be reliably estimated by reference solely to the pipette-manufacturer's specifications. Such estimates do not include external sources of variation and may be regarded as minima. The quality of estimation can be improved by using information gained by laboratory ('in-house') re-calibration of the pipettes. The degree of improvement was assessed by comparison with similar series made without re-calibration. It was found that improvement was minimal for colony counts but worthwhile for homogeneous solutions.

Horner's syndrome following excision of a vagal paraganglionoma.

We report a case of excision of a vagal paraganglionoma resulting in Horner's syndrome. The case was initially misdiagnosed as a carotid body tumour and demonstrates the need for adequate preoperative imaging and patient counselling for likely complications of surgery.

Severe panniculitis caused by homozygous ZZ alpha1-antitrypsin deficiency treated successfully with human purified enzyme (Prolastin).

Severe panniculitis caused by alpha1-antitrypsin deficiency is very rare even though the ZZ phenotype occurs in 1 : 3500 of the population of northern Europe. We describe a 33-year-old woman with rapidly progressing panniculitis and extensive skin necrosis with multiple life-threatening complications. Initial treatment followed by maintenance therapy with human purified enzyme (Prolastin, Bayer, Bridgend, U.K.) has been life-saving.

Estimating the precision of serial dilutions and viable bacterial counts.

The propagation of error in serial dilutions was investigated theoretically and by means of computer simulations. The principal aim of the study was, given only the pipette manufacturer's specification, to estimate the variance of any step in a dilution series both of pure solutions and of homogeneous bacterial suspensions by means of simple formulae. The study was extended to include bacterial plate counts by both the standard and the Miles and Misra methods. It was found that such estimation was possible and that the distributions approximated the normal sufficiently for the construction of confidence intervals (Cls) by the usual method. Such intervals can be regarded as minima which could be inflated by other, possibly undetermined, factors. It is suggested that laboratories could construct tables such as that reported here for pipettes and methods in common use to facilitate estimation. While replication of the final sampling step of a plate count increases the precision of estimation, averaging across dilutions may decrease precision and is not recommended for the standard pour-plate count.

Axillary artery pseudoaneurysm.

Axillary artery aneurysms are extremely rare. We report a case of an axillary artery aneurysm in a keen rugby player who had previous internal fixation of the shoulder for recurrent dislocation.

Antimicrobial activity of fluoroquinolone photodegradation products determined by parallel-line bioassay and high performance liquid chromatography.

The fluoroquinolones produce multiple photodegradation products. Little is known about these products, particularly whether any possess antimicrobial activity. To investigate this, we used the parallel-line bioassay to investigate discrepancies in zone of inhibition size in conjunction with high performance liquid chromatography (HPLC) analysis. A continuous flow photochemical reaction unit ('Beam-Boost') was used to partially photodegrade the fluoroquinolones ofloxacin, levofloxacin, ciprofloxacin and moxifloxacin (0.02 mM) by between 15 and 89%, as confirmed by HPLC. The concentration of residual parent fluoroquinolone in each irradiated sample was measured by HPLC and a non-irradiated control solution was prepared at the same concentration. These were compared by parallel-line bioassays using Escherichia coli, Enterobacter cloacae and Klebsiella oxytoca. With ofloxacin and levofloxacin, the zone size for the control solution was significantly less than that of the irradiated solutions, with >15% photodegradation in at least two of the indicator organisms, indicating that the photodegradation products possess antimicrobial activity. No difference was seen with ciprofloxacin at any level of photodegradation with any of the indicator organisms, nor with moxifloxacin at 30 and 54% photodegradation. A significant difference was observed with E. cloacae only, at 83% photodegradation.

Mortality from ruptured abdominal aortic aneurysm in Wales.

BACKGROUND: The aim of this study was to identify the incidence of, and mortality in, patients with a ruptured abdominal aortic aneurysm (AAA) reaching hospital alive in Wales. METHODS: Patients who presented with a ruptured AAA between September 1996 and August 1997 were analysed. Data were collected prospectively by an independent body, observing strict confidentiality. RESULTS: Some 233 patients with a confirmed ruptured AAA were identified, giving an incidence of eight per 100 000 total population. Some 133 patients (57 per cent) underwent attempted operative repair; 85 (64 per cent) of these died within 30 days. Of the 233 patients, 92 were admitted under the care of a vascular surgeon and 141 under a non-vascular surgeon. Vascular surgeons operated on 82 patients (89 per cent), of whom 50 (61 per cent) died, whereas non-vascular surgeons operated on 51 patients (36 per cent), of whom 35 (69 per cent) died. DISCUSSION: This study is unique as it is an independent prospective study of mortality in patients with a ruptured AAA who reached hospital alive. Mortality was independent of the operating surgeon, but vascular surgeons turned down significantly fewer patients than non-vascular surgeons (11 versus 64 per cent, P < 0.001).

Ideal pain relief following laparoscopic cholecystectomy.

In a previous report the effectiveness of intraperitoneal bupivacaine in reducing pain following laparoscopic cholecystectomy was demonstrated. Other methods of pain relief are commonly used but none has been compared following laparoscopic cholecystectomy. In two further studies we have compared the analgesic effect of intraperitoneal bupivacaine against wound infiltration with bupivacaine, and against intraperitoneal bupivacaine with the addition of a non-steroidal anti-inflammatory drug (NSAID) in patients undergoing laparoscopic cholecystectomy. Two consecutive studies were performed. In the first, patients in group 1 were given 20 ml of 0.25% bupivacaine into the peritoneal cavity; patients in group 2 were given 20 ml of 0.25% bupivacaine injected into the trocar wounds. In the second study, patients in group 1 were given 20 ml of 0.25% bupivacaine into the peritoneal cavity; patients in group 2 were given 20 ml of 0.25% bupivacaine into the peritoneal cavity and a diclofenac suppository (100 mg) one hour before surgery. Postoperative pain was assessed with a visual analogue pain scale. There was no difference in pain scores in the two groups in either study. Intraperitoneal bupivacaine is as effective as wound infiltration. The addition of an NSAID makes no difference in the reduction of postoperative pain following laparoscopic cholecystectomy.

The pharmacokinetics of intravenous ciprofloxacin 400 mg 12 hourly in patients with severe sepsis: the effect of renal function and intra-abdominal disease.

Serum concentrations of ciprofloxacin were reviewed in 22 patients given ciprofloxacin 400 mg intravenously 12 hourly for severe infection. No dosage modifications were made in patients with renal impairment. Patients who had either bowel or liver pathology in addition to renal failure had significantly higher serum concentrations than all other patients. Dosage reduction of ciprofloxacin in patients with severe sepsis and impaired renal function is not required unless they have co-existent intra-abdominal disease.

Is an outpatient appointment required for patients referred for a minor operation under local anaesthetic?

The necessity of an outpatient (OP) consultation for patients referred for minor surgery under local anaesthetic (LA) was examined. Two separate prospective studies were performed. The first comprised 107 patients referred for minor surgery, who were assessed in the outpatient department (OPD), before being booked for minor operations. The second study comprised 106 patients whose names were placed directly on a minor operations list on the basis of the GP referral letter alone. In the first study, 97 (91 per cent) patients went on to undergo minor surgery. Five were not suitable for LA and in five an operation was considered unnecessary. Patients not suitable for LA included children, neck swellings, pre-auricular swellings and swellings described as lymph nodes. In the second study, the GP referral letters were screened to exclude the above and of 106 referrals 93 (88 per cent) underwent a minor operation. The benefit of the second study was twofold. First, 106 OP slots were available for other referrals and secondly, patients avoided the OP waiting list and did not lose time from work as a result of the OP visit. There were no adverse effects demonstrated during the second study. We believe the OP consultation can be avoided if the referral letters are carefully screened.