Development of a PCR-free electrochemical point of care test for clinical detection of methicillin resistant Staphylococcus aureus (MRSA).

An MRSA assay requiring neither labeling nor amplification of target DNA has been developed. Sequence specific binding of fragments of bacterial genomic DNA is detected at femtomolar concentrations using electrochemical impedance spectroscopy (EIS). This has been achieved using systematic optimisation of probe chemistry (PNA self-assembled monolayer film on gold electrode), electrode film structure (the size and nature of the chemical spacer) and DNA fragmentation, as these are found to play an important role in assay performance. These sensitivity improvements allow the elimination of the PCR step and DNA labeling and facilitate the development of a simple and rapid point of care test for MRSA. Assay performance is then evaluated and specific direct detection of the MRSA diagnostic mecA gene from genomic DNA, extracted directly from bacteria without further treatment is demonstrated for bacteria spiked into saline (10(6) cells per mL) on gold macrodisc electrodes and into human wound fluid (10(4) cells per mL) on screen printed gold electrodes. The latter detection level is particularly relevant to clinical requirements and point of care testing where the general threshold for considering a wound to be infected is 10(5) cells per mL. By eliminating the PCR step typically employed in nucleic acid assays, using screen printed electrodes and achieving sequence specific discrimination under ambient conditions, the test is extremely simple to design and engineer. In combination with a time to result of a few minutes this means the assay is well placed for use in point of care testing.

Spray drying of budesonide, formoterol fumarate and their composites--I. Physicochemical characterisation.

The objective of this work was to examine the physicochemical properties of spray dried budesonide, formoterol fumarate and their mixtures at two different weight ratios: 100:6 and 400:6 of budesonide and formoterol fumarate, respectively. A comparison of the thermal properties, crystalline/amorphous nature and particle size of the starting micronised as well as processed materials was carried out. The micronised drugs on their own and the physical mixtures were crystalline in contrast to the spray dried counterparts which were shown to be amorphous. The glass transition temperatures (T(g)s) of the processed actives were determined and appeared at 89.5 and 88 degrees C for budesonide and formoterol fumarate, respectively. As for the spray dried composites, an indication of miscibility and/or interactions between the components was indicated by differential scanning calorimetry and infrared analysis. The spray drying in all cases resulted in smooth, spherical microparticles of sizes suitable for inhalation.

Pilot study of objective cough monitoring in infants.

Cough is common in childhood, resulting in significant morbidity and frequent medical consultation. Despite this, little is known about the frequency or development of cough, particularly in infants and young children. Recent progress in monitoring has enabled cough to be measured objectively both day and night. However, to date, objective measurement has only been used in adults and older children. The aim of this study was to see whether such methods could be extended to allow objective cough monitoring in infants. Thirty infants were recruited: 13 with coughing illnesses (group 1), and 17 normal, healthy babies (group 2) born to nonatopic, nonsmoking parents. Group 2 infants were studied when well, several times in the first year of life. Coughs were recorded using an adapted commercial cough monitor (Logan Research LR100) and simultaneous infrared video and sound recording. Thirty-eight recordings with simultaneous cough monitor and video data were analyzed: 9 from group 1, and 29 from group 2. Overall, the sensitivity of the monitor when compared to video was 81%, with a positive predictive value of 0.8. There was good agreement between the two methods for infants with infrequent cough (<5 coughs per hour). Agreement in infants with more frequent cough was not as good, because more coughs were consistently identified by the cough monitor. The portability and small size of the cough monitor made it easy to use, although there were difficulties in keeping the leads attached in older, more mobile infants. In conclusion, objective assessment of cough is practical in infants, enabling the pattern of cough in illness and in health to be studied further.

Development of a bioassay for phytochemicals using Daphnia pulex.

The freshwater cladoceran Daphnia pulex was explored as an alternative to Artemia salina for the biological screening of phytochemicals. This paper reports on the results of screening 27 compounds, and comparisons are made with screens using Artemia salina. The effect of miniaturisation of the assay system was investigated. Petri-dish based tests were performed for the 27 compounds using 10 daphnids at each of 5 concentrations (1 - 1000 mg l(-1)). Potassium dichromate was used as a control and the number immobile after 24 hours were counted. Results were expressed as EC(50) values. The active compounds (EC(50) < 25 mg l(-1)) comprised mainly adrenoceptor agonist alkaloids, while the cytotoxins vincristine and colchicine, and several antibiotics were relatively non-toxic towards Daphnia. The method was validated by comparative bioassay of A. belladonna fractions. Overall, this screen compared favourably with those based on Artemia. Daphnia were sensitive to a wide range of biologically active molecules including CNS-stimulants, anti-malarials, narcotics and anti-spasmodics, and the sensitivity was broader than previously seen with Artemia. The screen is perhaps limited by the lack of sensitivity to cytotoxins, but this might be overcome by altering the test criteria.

Relationship between knee extension force and stand-up performance in community-dwelling elderly women.

OBJECTIVE: To describe the relationship between knee extension force and stand-up performance. DESIGN: Prospective, cross-sectional, descriptive, and correlational. SETTING: Community. PARTICIPANTS: Fifty-five community-dwelling women 60 or more years old. INTERVENTIONS: Information pertaining to stand-up performance was gathered via structured questions. The time to complete 1 sit-to-stand from a bench of known height (16in) was obtained in conjunction with subjects' ratings of perceived exertion (RPEs) relative to the task. Bilateral lower extremity knee extension strength was measured with a hand-held dynamometer and normalized against body weight. MAIN OUTCOME MEASURES: Total knee extension forces (raw and body-weight normalized values) generated by each subject were correlated with 3 measures of stand-up performance. RESULTS: Low to moderate correlations (r = -.323 to -.526) were found between knee extension forces and the stand-up performance measures of bench stand time and stand-up difficulties. Normalized knee extension force had a higher correlation than raw knee extension force with stand-up difficulties and RPE. Correlations were higher when the curvilinear nature of the relationship was taken into account. The highest correlation was between normalized knee extension force and the RPE associated with the bench stand task (r = -.767). CONCLUSION: Knee extension force offers an important but incomplete explanation of stand-up performance from low surfaces by elderly women. Interventions leading to increased knee extension force may improve stand-up performance.

Phenylalkylamine abuse among opiate addicts attending a methadone treatment programme in the Republic of Ireland.

Since the early 1990s ring-substituted derivatives of amphetamine have been abused widely in the Republic of Ireland. The main ring-substituted amphetamines being abused include methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA) and methylenedioxyethamphetamine (MDEA). A newer illicit synthetic analogue, which has been seized to a lesser extent by Irish police, is N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB). The work presented here involved the determination of the type of ring-substituted amphetamines being abused by a group of recovering opiate abusers participating in a methadone maintenance programme in a Dublin Drug Rehabilitation Centre. Urine samples which tested positive for amphetamines and ring-substituted amphetamines via EMIT immunoassay were subjected to further analysis using GC-MS with MBTFA flash derivatization. It was found that the methylenedioxypropanamines were being abused, as was amphetamine itself. However, no abuse of methylenedioxybutanamines or thioamphetamines was observed.

A broad range of forces is encompassed by the maximum manual muscle test grade of five.

This retrospective study summarizes the range of knee extension forces associated with the manual muscle test grade of 5. The range of the forces (>560 Newtons) was large and encompassing of more than 86% of measurable forces. These findings help to explain the insensitivity of manual muscle testing and why it has such a profound ceiling effect.

Tuberculosis in children at the Royal Hospital for Sick Children, Glasgow. January 1995-April 1998.

BACKGROUND: The management of paediatric tuberculosis can be complicated. Since 1995, children referred to the Royal Hospital for Sick Children have been managed at a specialist tuberculosis clinic, in an attempt to standardise diagnosis, treatment and follow-up. AIMS: To review the management of paediatric tuberculosis at the Royal Hospital for Sick Children, Glasgow, from January 1995 to April 1998. METHODS: Retrospective study using information from the management database of all children treated for tuberculosis since January 1995. RESULTS: Seventy two children have been treated (median age 5.9 [range 2.2-9.2] years; sex male:female 40:32; 76% Caucasians): 43 for tuberculosis disease, 21 for tuberculosis infection (chemoprophylaxis) and eight as tuberculosis contacts. Sixty three per cent were identified by contact tracing eighty one per cent of Mantoux tests were documented accurately. Of those with disease, 93% had gastric washings taken, and positive microbiological confirmation was obtained in 33%. All patients with disease were commenced on the chemotherapy regimen recommended by the British Thoracic Society. 88% completed treatment. Eighty four per cent of those with disease had been notified. Treatment of those in the infection or contact groups were treated more aggressively than the British Thoracic Society guidelines, with six months of treatment with isoniazid and rifampicin. Directly observed therapy was used in 17 children (24%). Only 53% of Indo-asian children had received BCG immunisation. CONCLUSIONS: Most children were managed according to the British Thoracic Society guidelines. A management system, with a computerised database, has facilitated both patient management and audit, and ensured the delivery of quality care.

Hepatic enzyme abnormalities in children on triple therapy for tuberculosis.

Standard chemotherapy for tuberculosis (TB) in children uses hepatotoxic drugs. Published data and guidelines on monitoring of liver function during TB treatment are often contradictory and not directly relevant to the pediatric population. We carefully monitored 43 children (age 6.6 years, 0.7-15.1 [median, range]; 49% male; 72% Caucasian) being treated for TB infection (n = 8) or disease (n = 35) with triple therapy, using pyrazinamide, rifampicin, and isoniazid in standard recommended doses. Children on other hepatotoxic drugs were excluded. Measurements of liver function tests (LFT) included aspartate transaminase (AST), alanine transaminase (ALT), and bilirubin, and they were checked before and a median of 5 times (1-23) during treatment. Only one child had mildly abnormal LFTs pretreatment. Thirteen children (n = 13, [30%]; age 7.6 years, 1.8-10.9; 54% male; 77% Caucasian) developed abnormal LFTs (> mean + 2 SD) and of these 10 had TB disease. Eight of the 13 had mildly elevated enzymes (< twice upper limit of normal) while in five, all with disease, the enzymes were more markedly raised. In the group with normal LFTs (n = 30, [70%]; age 6.6 years 0.7-15.1; 47% male; 70% Caucasian) 25 had disease (83%). Liver enzyme elevation occurred early (1.65 weeks, 0.6-16.6). Only two children had symptoms (one jaundice, one pruritus) with treatment being stopped temporarily only in the jaundiced child. Otherwise, LFTs normalized without interrupting treatment. We conclude that elevated liver enzymes are not uncommon in children receiving triple therapy for TB, generally occurring early in treatment. Symptoms are rare. Current British Thoracic Society and American Thoracic Society guidelines (that if LFTs are normal prior to treatment then further monitoring should only be performed if clinically indicated) seem adequate for children.

Experimental measurements of sidebranching in thermal dendrites under terrestrial-gravity and microgravity conditions.

We perform sidebranch measurements on pure succinonitrile dendrites grown in both microgravity and terrestrial-gravity conditions for a set of supercoolings within the range 0.1-1.0 K. Two distinct types of sidebranch regions, uniform and coarsening, exist, and are characterized by the distance from the tip at which the region began, D(i), and the average spacing of sidebranches within that region, lambda(i). There does not appear to be any significant dependence on either gravity level or supercooling when D(i) or lambda(i) are normalized with respect to the radius of curvature of the tip, R. The apparently constant normalized proportionality factor between D(i), lambda(i), and R, regardless of the relative importance of diffusion and convective heat transport, demonstrates self-similarity between dendrites of different length scales propagating under various heat transfer conditions. However, when the form of the sidebranch envelope is defined by a power law relating the amplitude and relative positions of the sidebranches normalized to the radius of the tip, the form is seen to have significant variations with supercooling between the terrestrial gravity and microgravity growth dendrites. Furthermore, both the amplitude coefficient and exponent from the power-law regressions of the microgravity data are statistically different (95% confidence level) than their terrestrial counterparts.

Comparison of resistance of fungal spores to gamma and electron beam radiation.

The irradiation sensitivity of fungal spores to either gamma or electron beam irradiation was evaluated in distilled water. The D10 (the dose required to reduce the initial population by 90%) gamma values ranged from 0.236 to 0.416 kGy and from 0.209 to 0.319 kGy for Penicillium and Aspergillus species, respectively. The D10 electron beam values ranged from 0.194 to 0.341 and from 0.198 to 0.243 kGy for Penicillium and Aspergillus species, respectively. Of the aspergilli species evaluated, only half exhibited significantly (P < 0.05) greater sensitivity to the electron beam treatment compared to gamma irradiation. Four of the six penicillia species evaluated also exhibited significantly (P < 0.05) higher sensitivities to electron beam treatment.

The effects of aerobic and anaerobic exercise conditioning on resting metabolic rate and the thermic effect of a meal.

This study examined resting metabolic rate (RMR) and thermic effect of a meal (TEM) among athletes who had participated in long-term anaerobic or aerobic exercise. Nine collegiate wrestlers were matched for age, weight, and fat-free weight with 9 collegiate swimmers. Preliminary testing included maximal oxygen consumption, maximal anaerobic capacity (MAnC) for both the arms and the legs, and percent body fat. On two separate occasions, RMR and TEM were measured using indirect calorimetry. VO2max was significantly higher in the swimmers while MAnC was significantly higher in the wrestlers for both the arms and the legs. RMR adjusted for fat-free weight was not significantly different between groups. The differences in total and percentage of TEM between the groups were not statistically significant, and there were no differences in baseline thyroid hormones. These data suggest that despite significant differences in VO2max and WAnT values following long-term aerobic and anaerobic exercise training, resting energy expenditure does not differ between these college athletes.

Two seasons of weight cycling does not lower resting metabolic rate in college wrestlers.

Over the course of 2 yr, we prospectively studied the effect on resting metabolic rate (RMR) of multiple cycles of weight loss followed by regain in six weight cycling collegiate wrestlers (WC) (mean age = 19.0 yr) compared with 12 weight stable, physically active, nonwrestling controls (C) (mean age = 20.9 yr), whose body composition was similar to the wrestlers. Furthermore, during the second year of the investigation, a group of six nonweight cycling collegiate wrestlers (NWC) (mean age = 18.8 yr) were included in the analyses. The WC had previously undergone at least three seasons of weight cycling and continued this pattern during each year of the study. For the WC, RMR was determined by indirect calorimetry before and after a 6-month season of weight cycling for each of two consecutive years. A similar time frame was followed for measurement of RMR in the C, while for the NWC, pre- and postseason RMRs were measured only during the second year. During the 2 yr, the WC had significantly higher (P < 0.05) pre- and postseason measures of RMR compared with the C. A separate analysis comparing all three groups during the second year showed that RMR was not different for WC and NWC, and that RMR was higher for both wrestling groups compared with C. In this prospective study, weight cycling did not increase the thermogenic efficiency of collegiate wrestlers compared with either nonweight cycling wrestlers or weight stable controls, who were not wrestlers.

Hyperparathyroid crisis and posterior mediastinal parathyroid adenoma: a case for preoperative localization.

Hyperparathyroid crisis is a rare, life-threatening condition that requires prompt removal of the involved parathyroid gland(s). This report describes a patient with hyperparathyroid crisis caused by a mediastinal parathyroid adenoma, which was localized before operation and removed at the initial operation, without neck exploration. Surgical approaches to hyperparathyroid crisis and the importance of preoperative localization studies are discussed.

Gross efficiency responses to exercise conditioning in adult males of various ages.

This study investigated gross efficiency changes in a group of 60 adult males (mean age 39.2 +/- 1.2 years) resulting from endurance training and age-related responses to such training in sub-groups (each n = 20) of younger (30.7 +/- 0.7 years), intermediate (38.3 +/- 0.5 years) and older (48.6 +/- 1.1 years) subjects. Gross efficiency (%) was calculated from work output, oxygen consumption and RER energy equivalents following 10 min standard cycle ergometry exercise at 100 W and 50 rev min-1. Measurements were made at pre-, mid- and post-8 months of training, which involved progressive walking/jogging activities designed to enhance endurance capacity. In the total group, VO2 decreased pre- to post-training from 2.15 +/- 0.02 to 1.93 +/- 0.01 1 min-1 (P less than 0.01). In the sub-groups, both the younger and older subjects showed a significantly reduced VO2, from 2.17 +/- 0.01 to 1.98 +/- 0.04 1 min-1 and 2.05 +/- 0.08 to 1.86 +/- 0.03 1 min-1 respectively (P less than 0.05), but no significant changes were noted at mid-training. In the intermediate age subjects, while there were trends towards a reduced VO2, none was significant. The ANOVA revealed increased mean gross efficiency in the total group from pre- (14.3 +/- 0.1%) to post- (15.5 +/- 0.2%) (P less than 0.05) but not at mid-training (14.8 +/- 0.2%). While similar trends were observed in the sub-groups, gross efficiency increases were not significant, although changes in gross efficiency were reflected in VO2. The findings suggest that during standardized exercise, oxygen cost may be reduced and gross efficiency increased in adult males following endurance training and that such changes may take place over a variety of age ranges.

Gynaecological day surgery at an area hospital 1983-1987.

An eight bedded self-contained day unit for minor gynaecological surgery was opened in November 1982 at Craigavon Area Hospital. In the five years from 1 January 1983 to 31 December 1987, 2091 minor procedures have been performed in the unit, or 32.85% of the total gynaecological operations. The waiting time from consultation until admission for minor surgery was reduced from 2-3 months in 1983 to 2-3 weeks in 1987 and major surgery from 5-6 months to 1-2 months. Allied to this improvement in patient service has been an increase in the in-patient major surgery operation rate, resulting in 100 extra major operations per year. Although costs of treatment in the day procedure unit were lower than the average cost for a 24 hour in-patient stay, the cost per patient is dependent upon the total patient throughput of the unit and the number of days it is used. Complication rates (ie transfer to in-patient facilities for complications of surgery or anaesthesia) are low at 1.85%.

Resting metabolic rate in weight-cycling collegiate wrestlers compared with physically active, noncycling control subjects.

To determine the effect of multiple cycles of weight loss and regain on resting metabolic rate (RMR), we compared RMR between 12 weight-cycling collegiate wrestlers (means age 19.4 y) with a minimum of three previous seasons of weight cycling and 13 weight-stable nonwrestlers of similar weight and body composition (means age 20.6 y). RMR was measured before, during, and after a 6-mo wrestling season. Wrestlers exhibited a significantly higher baseline RMR compared with the control subjects (p less than 0.05). During the season when wrestlers had lost weight for competition, RMR was reduced (p less than 0.05) but was not significantly lower than that of the weight-stable control subjects. After a season of weight cycling and the final weight regain, the wrestler's postseason RMR was similar to preseason values and higher than the postseason RMR of the control subjects (p less than 0.05). Participation in numerous cycles of weight loss and regain did not lower RMR in these competitive athletes, as has been previously suggested.