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The influence of experimental protocol on estimation of adherence of P. aeruginosa to buccal epithelial cells is highlighted. Use of membrane filtration to remove non-adherent bacteria was significantly affected by membrane pore size and rinse volume, but was not affected by inoculum size or test isolate. Even with optimum filtration conditions, over 58% of the non-adherent bacterial population were retained on the filters. Separation of non-adherent bacteria was greatly improved when density centrifugation was employed, with less than 30% of non-adherent bacteria present in the buccal epithelial suspension. Adherence values measured by a radiological and fluorometric method were significantly higher than those determined by microscopic counting. Radiological assays had the lowest variance. Gradient centrifugation was used to prepare a standard suspension of buccal epithelial cells. However, cells collected on different days showed significantly different adherence measurements. Adherence of bacteria to trypsinized buccal epithelial cells and to buccal epithelial cells from cystic fibrosis patients was similar, whereas adherence to normal buccal epithelial cells was significantly lower.

Arthroconidia of Trichophyton mentagrophytes demonstrated an exogenous type of dormancy under environmental conditions unsuitable for germination, provided that these conditions were not lethal. Germination did not occur at 4 degrees C unless arthroconidia were shifted up to 37 degrees C. At 45 degrees C germination was inhibited even when arthroconidia were shifted down to 37 degrees C. Germination was dependent on humidified incubation conditions. The presence of human corneocytes enhanced the germination of arthroconidia in the presence of moisture.

The emergence of germ tubes from arthroconidia of Trichophyton mentagrophytes on stripped sheets of stratum corneum from different body areas was measured. Arthroconidia increased in size and started germination by 4 h at 37 degrees C. Germ tubes originated from a point on the arthroconidium surface mid-way between the points of attachment to adjacent conidia. With further incubation arthroconidial germination increased and germ tubes extended across the stratum corneum. Histological staining of transverse sections of infected stratum corneum showed hyphae penetrating longitudinally and perpendicularly through the thickness of the stratum corneum. By 7 days' incubation hyphae started to form arthroconidia thereby completing the vegetative growth cycle of the fungus. Scanning electron microscopy revealed penetration of corneocytes by germ tubes resulting in damage to the corneocyte surface.

This longitudinal study identified processes of exercise behaviour change (POC) associated with movement between the stages of exercise behaviour change (SOC). Participants’ ( N = 312) physical activity, SOC and POC were recorded at baseline, one, three, six and 12 months post-test. Following baseline, participants received one of three physical activity interventions. The process of self-liberation was important at each stage movement. The process of stimulus control appeared important when progressing from contemplation to preparation. Progression from action into maintenance was associated with increased use of social liberation and helping relationships. The study indicated the transtheoretical model is applicable to a British population.

The objectives were to establish the origin of 2 acute phase proteins in milk during subclinical bovine mastitis and to characterize the relationship between those proteins in milk and blood. Haptoglobin (Hp) and mammary-associated serum amyloid A (M-SAA3) appear in milk during mastitis, whereas Hp and serum amyloid A increase in serum during mastitis. The concentrations of these proteins were determined in an experimental model using a field strain of Staphylococcus aureus to induce subclinical mastitis in dairy cows. The expression of mRNA coding for these proteins was assessed and the presence of M-SAA3 in mammary tissues was determined using immunocytochemistry. Increases of M-SAA3 and Hp in milk occurred within 12 h of Staphylococcus aureus infusion, with peak concentrations occurring 3 d after infusion of the bacteria. The increase of acute phase proteins in milk (15 h) preceded the increase in serum concentrations of both proteins (24 h). Expression of mRNA for M-SAA3 and Hp increased in both mammary and hepatic tissues 48 h after infusion of the mammary glands. In mammary tissue, the increase of M-SAA3 mRNA was greater than the increase in Hp mRNA expression, whereas in hepatic tissue, the increase in M-SAA3 mRNA was less than that for Hp mRNA. Immunocytochemistry demonstrated that M-SAA3 protein was present within secretory epithelial cells at significantly higher levels in infected mammary glands than in control tissues. These proteins, which have host defense and antibacterial activities, may play a significant role in the early response to invasion of mammary tissues by pathogenic bacteria.

Airborne gamma spectrometry (AGS) is being increasingly recognised as an important means for mapping environmental radioactivity in emergency response. Progress has been made in recent years towards methodological convergence and cooperation between European teams. Recently, an international comparison was undertaken in SW Scotland in 2002 to evaluate AGS and ground-based methods. Teams from 18 institutions in 10 European countries attended, collecting some 140,000 AGS spectra, with 750 laboratory gamma spectrometry analyses and 120 in situ observations from the ground sites. Comparisons between AGS and ground-based methods have confirmed the validity of AGS protocols. A composite mapping task, where AGS teams recorded data over adjacent parts of a 90 x 40 km 2 area within a few days, confirmed the ability of teams to work together in an effective manner. This paper provides a summary of the results of the exercise. These demonstrate the operational capabilities of European AGS teams and confirm the quantitative nature of the method.

The climate shift towards wetter conditions at the transition from Subboreal to Subatlantic in NW-Europe (ca 850 cal. yrs BC; caused by a decline of solar activity), is also evident in South Siberia. Areas that initially were hostile semi-deserts changed into attractive steppe landscapes with a high biomass production, and therefore high carrying capacity. We focus on south-central Siberia where an acceleration of cultural development and an increase in the density of nomadic Scythian populations took place shortly after 850 BC. We hypothesize a causal relationship between the Scythian expansion and migration, and the early Subatlantic shift towards increased humidity.

An in vitro method of determining the activity of antibiotics in combination which is simple and convenient to perform and which could be used routinely in clinical microbiology laboratories is desirable. We investigated the activity, against Pseudomonas aeruginosa and Burkholderia cepacia complex clinical isolates, of ceftazidime and tobramycin in combination using a broth macrodilution sensitivity method based on breakpoint minimum inhibitory concentrations and compared the results obtained using this method with those obtained using the microtitre checkerboard method. There was good agreement in interpretation of results between the two methods for both P. aeruginosa (90%) and B. cepacia complex isolates (70%) with tobramycin and for P. aeruginosa isolates (70%) with ceftazidime. As the breakpoint combination sensitivity testing method employs only four tubes and does not require initial determination of individual antibiotic minimum inhibitory concentrations, it is simpler and more convenient for determining the activity of antibiotics in combination than the microtitre checkerboard method. The use of this method in routine microbiology laboratories to determine the activity of antibiotic combinations against clinical isolates should optimise treatment of infection by ensuring that appropriate antibiotic combinations are prescribed.

Stimulation of long lasting, protective immunity to respiratory viruses is often difficult to achieve with conventional respiratory vaccines. Polymeric nanoparticles, incorporating viral proteins have been shown to offer sustained release of antigen, with consequent prolongued stimulation of the respiratory immune system. In this paper the efficacy of two nanoparticle vaccines (poly-lactide-co-glycolide, PLGA; polymethylmethacrylate, PMMA), incorporating proteins of bovine parainfluenza type 3 virus (BPI-3) was investigated. As a preliminary to experiments in calves, it was considered essential to demonstrate immunogenicity of the experimental vaccine in mice. Mice immunised with PLGA nanoparticles, containing BPI-3 proteins, developed higher levels of virus-specific antibody than mice immunised with the PMMA vaccine or with soluble viral proteins alone. Immunoblotting using serum from the vaccinated mice, demonstrated strong reactions against the major BPI-3 proteins.

Background/aims Tail docking neonatal piglets remains a controversial animal welfare issue. Although banned in the UK, it is widely practiced in many countries as a safeguard against tail biting among pigs reared in intensive systems. Concerns exist whether tail docking can induce chronic pain in later life. This preliminary study examined the effects of partial tail amputation on activating transcription factor 3 (ATF3), a marker of peripheral nerve injury and regeneration and NMDA-glutamate receptor NR2B subunit which participates in the mediation of chronic pain. Methods Procedures were performed according to the ethical guidelines for the study of experimental pain in animals. Six piglets (2–3 days old) were tail-docked (a portion of the tail amputated with sterile surgical cutters), six piglets (2–3 days old) were sham-docked. Three animals from each treatment were euthanised 7 and 56 days post-amputation. Coccygeal dorsal root ganglia (DRG) and spinal cord were collected post-mortem for immunohistochemistry. Results ATF3 immunoreactivity (IR) was significantly increased (p p < 0.05) in dorsal horn neurons in tail-docked piglets compared with intact piglets 7 days after docking. There was no difference in NR2B-IR in neurons 56 days post amputation, compared with intact piglets. Conclusions Increased ATF3 and NR2B-IR 7 days after tail-docking suggests that injury to the peripheral nerves in the tail was sufficient to trigger neuronal regeneration and altered dorsal horn signaling respectively, however the effects of tail-docking on neuronal regeneration and nociceptive signaling were relatively short lasting. Tail-docking neonatal piglets does not cause sustained changes in ATF3, which might suggest ongoing nerve fibre damage and NR2B which might be implicated in chronic pain.

The main aim of this study was to assess the effects of a fitness assessment and exercise consultation on physical activity over 1 year in non-regularly active participants drawn from a socially and economically deprived community. Of 3000 people invited to volunteer for either intervention, 225 fitness assessment volunteers were randomly assigned to an experimental or control group; 145 exercise consultation volunteers were similarly assigned. Physical activity was measured at baseline, 4 weeks, 3 months (plus an intervention re-test), 6 months and 1 year. Analysis of variance and follow-up Bonferroni analysis showed that, for those not regularly active at baseline, physical activity increased significantly to 4 weeks, was maintained to 6 months but had fallen by 1 year. Only those receiving an exercise consultation significantly increased their physical activity after 1 year. Compared with fitness assessments, chi-square analysis showed that significantly more non-regularly active participants volunteered for an exercise consultation and those receiving an exercise consultation had significantly better long-term study adherence than those receiving a fitness assessment. The study also showed that, contrary to popular opinion, those in a socially and economically deprived community are not 'hard to reach' and respond well to physical activity interventions.

A composite scale for assessing pain in dogs in a hospital setting has been developed on the basis of observations of their behaviour. Initially, 279 words and expressions suggested by 69 veterinary surgeons were reduced into 47 words and expressions which were allocated into seven behaviour categories: demeanour and response to people, posture, mobility, activity, response to touch, attention to painful area and vocalisation. Three statistical methods, hierarchical agglomerative cluster analysis, Cronbach's alpha coefficient, and analysis of variance with multiple comparisons and empirical cumulative distributions, were used to validate these procedures, and a questionnaire accompanied by a list of definitions was designed around the expressions. The new composite scale is more detailed than previously reported scales for assessing pain in dogs on the basis of their behaviour, and the methods used in its development are based on sound scientific principles.

An innovative approach using a multiple-study research programme that incorporated five individual projects, each of which can be interpreted independently or as an integral part of the whole

OBJECTIVE: To determine the quality of life of elderly patients with osteoarthritis (OA) compared with that of their peers with no chronic illnesses and to investigate the associations between analgesic use and quality of life. SUBJECTS: Patients >65 years of age with OA taking analgesics with (n = 33) and without (n = 26) comorbidities, and control patients with no chronic illness and not taking analgesics (n = 37). METHODS: Quality of life was assessed by the SF-36 and level of pain was measured by a visual analog scale. Multiple regression analysis was used to determine the medication characteristics and additional patient factors significantly associated with SF-36 scores. RESULTS: OA patients had significantly (p < 0.05) lower scores than control patients in all quality-of-life domains. OA patient scores were lowest for the domains of role-physical, bodily pain, and physical functioning. OA patients with comorbidities also had poorer general and mental health. Pain- and analgesic-related factors were significantly (p < 0.05) associated with physical health status, reduced vitality, general health, and social functioning. A better quality of life was associated with noncompliance, fewer visits to the physician, and taking oral nonsteroidal antiinflammatory agents. Presence of adverse drug reactions and sleep disturbance did not influence SF-36 scores significantly. CONCLUSIONS: Quality of life of elderly OA patients with and without additional comorbidities was significantly poorer than that of their healthy peers, particularly in the domains associated with physical status, but also affecting vitality, social functioning, and general health. Level of pain suffered and perceived effectiveness of analgesic medication in pain control were important factors associated with quality of life.

Traditionally, sensitivity and uncertainty analyses in environmental modelling contribute quantitative descriptions of the relative importances of individual parameters and processes, highlighting areas of significant contributors to the overall uncertainties of model predictions and giving markers for areas requiring substantial improvement (perhaps through directed experimental work). Both sensitivity and uncertainty analysis require extensive re-sampling from input data and simulation of model response and there is a large and growing literature concerning their use in environmental modelling. There is however growing interest in the sensitivity of predictions to model structure, and in evaluating the contribution of model structural uncertainty to the overall uncertainty within the general framework of sensitivity analysis (Draper, 1995, Beven, 1993, Beven and Binley, 1992). Sensitivity and uncertainty analysis contribute to all stages of model development, testing and assessment and their impact on model reliability and validity will be described. After a general introduction to sensitivity and uncertainty analysis and discussion of their contributions, some applications of sensitivity analysis to several environmental modelling studies will be presented.

A new in vitro model for the study of nail invasion by dermatophyte fungi was developed. The dermatophyte Trichophyton mentagrophytes, and fragments of finger-nails and toe-nails were used. Arthroconidia were inoculated on the ventral surface of the nails. After 6 h, adherence and germination of arthroconidia was observed. By 16 h, small germ tubes with side branches were evident. At about 24 h, micro-colonies had become established. At 48 h, a mycelium had formed, and at about 72 h most of the nail fragment was covered with fungal growth. Nail penetration occurred from the ventral surface through the intercellular spaces, and with longer incubation all three layers were invaded by arthroconidia growing through channels. Nail invasion occurred in the absence of added nutrients. Dermatophyte fungi appeared to invade the nail by a combination of mechanical and chemical factors. The model provides a substrate to study the pharmacokinetics and bioavailability of new antifungal agents in situ.

Time-survival studies were conducted to estimate the effects of azlocillin and tobramycin on Pseudomonas aeruginosa NCIMB 8295 (in the exponential phase of growth) at concentrations ranging from one-quarter to twice the MIC. The effects of the individual agents and their combinations were determined by measuring the viable counts (CFU per milliliter) over a 24-h period. The typical pattern observed from the plot of the logarithm of the CFU per milliliter against time was an initial rapid killing; this was followed by a period of stasis and regrowth. Initial rates of killing by tobramycin were concentration dependent, whereas this was not the case with azlocillin. From the time-survivor plots, the area under the curve for viable bacteria was also calculated. It offered a useful method of interpreting the results of time-kill studies, taking the overall pattern of killing and regrowth into consideration. The area under the curve for viable bacteria was concentration dependent for both antibiotics. A 2(2) factorial experimental design was used to analyze the joint effects of azlocillin and tobramycin. In such a factorial experiment, an interaction between two factors, in this case, azlocillin and tobramycin concentrations, is shown by a change in the slope of the plot when the concentration of the interactant is changed. Analysis of variance showed that the combination was synergistic at low concentrations, but this was not significant when the concentration of either interactant was increased.

The following bibliography was compiled as part of a European Concerted Action on environmental airborne gamma-spectrometry (Project F14P-CT95-0017) conducted between 1996 and 1999 under the Fourth Framework Nuclear Fission Safety Research Programme. Between the time of the Chernobyl reactor accident in 1986, and the end of the project, a significant number of European groups have made developments in the radiometrics area, and have conducted environmental surveys. In addition to publications in open or peer-reviewed literature, there is a substantial body of material in the form of technical reports, which are for the most available from the originators on request. The bibliography reproduced here comprises titles and citation references to the material presented to the EU concerted action by the European AGS teams which were active during the study period. It is deliberately intended to be an inclusive resource rather than a selective one, which defines the range and scope of environmental AGS. An extended form of this information, including concise abstracts for many items, is included within the final report of project F14P-CT95-0017.

D. GILLILAND, A. LI WAN PO AND E. SCOTT. 1992. The antimicrobial effects of methyl and propyl parabens are investigated, with Escherichia coli as test organism, with a view to determining whether the parabens act synergistically. At appropriate concentrations, the parabens killed E. coli cells according to first order kinetics and the bactericidal effects were quantified by the first order kill rate constants. Combinations of methyl or propyl parabens, at concentrations which slow down or inhibit bacterial growth when used singly, produced definite kill. In this sense, the parabens are therefore synergistic since in combination they produce an effect which is not observed when they are used singly. This effect is not true synergism as shown by the results of our experiments with a factorial design. Analysis of variance indicated no significant interaction between the two parabens.

The effect of temperature on the kill rate of Escherichia coli by methyl and propyl parabens was studied. The kill kinetics was first order. It was shown that the Arrhenius equation provided a good model for describing the relationship between the first order rate constant and the temperature. The activation energy was found to be 274 kJ/mol for exponential phase cells and 168 kJ/mol for stationary phase cells. Exponential phase cells were much more susceptible to the lethal effects of the parabens than were the stationary phase cells. For example, at 34 degrees C stationary phase cells, in chemically defined media, had a kill rate constant of 0.072/h while the corresponding value for exponential phase cells was 0.238/h. In water the rate of kill for exponential phase cells was even faster giving a rate constant of 5.25/h at 34 degrees C. Non-isothermal kinetic testing was not found to be useful for modelling bacterial kill kinetics because we could not achieve the precision required in bacterial enumeration.

Five serial exposures of mucoid Pseudomonas aeruginosa from patients with cystic fibrosis to subinhibitory concentrations of ciprofloxacin resulted in stepwise increases in the MIC, with a mean proportional increase of 10. MICs were significantly lower in an iron-limited chemically defined medium than in Iso-Sensitest broth. The mucoid phenotype was maintained in chemically defined medium. Acquired resistance was retained either partially or completely in 85% of the isolates following 10 transfers in drug-free media. In cases in which susceptibility was regained, an increase in the MIC was observed on one further exposure to ciprofloxacin.

Focal points □ A self-selected set of pharmacists participated in a formal women's health care advice programme □ The tools (continuing education needs assessment grid and job competency wheels) proved useful in identifying training needs and assisted the implementation of identified training solutions □ Regular meetings of the set were an essential component of the process □ Motivation and encouragement by peers should not be underestimated □ Action learning in association with an organised programme is a successful and appropriate means of facilitating CPD

Focal points □ The pilot study evaluated the outcome of a pharmacy-based initiative for women starting or having recently started HRT □ At the end of the study over twice as many women in the control group had stopped taking HRT than in the intervention group □ Kupperman menopausal index scores used to indicate control of menopausal symptoms had improved significantly more in the intervention group compared with the control group □ Intervention group patients were more knowledgeable of the benefits of HRT, and of osteoporosis risk factors and preventive measures □ The control group expressed significantly more concerns about HRT at the end of the study compared with the intervention group

The emergence and development of chemical and thermal resistance in spores of Bacillus subtilis was examined. The chemicals studied were of the disinfectant type: glutaraldehyde, hypochlorite, hypochlorite-methanol and povidone-iodine. Growth and sporulation were followed by electron microscopy and resistance assigned to specific stages in relation to 45Ca and DPA accumulation. A sequential development of resistance was observed with thermal resistance appearing first at early Stage V corresponding to maturation of cortex and deposition of rudimentary spore coat material. Chemical resistance coincided with middle to late Stage V dependent on the chemical concerned. A progressive development of resistance was observed on prolonged incubation in sporulation medium and was affected by inclusion of lysozyme in the spore washing sequence.

SummaryIf Athabaskan Indians are subdivided by linguistic group, a wide diversity in gene frequencies is disclosed. This diversity approximated that found when linguistically unrelated groups were compared. It was greater than that found for Eskimo-Aleuts, even though the latter are more heterogeneous linguistically and subject to a wider variety of environmental conditions. Contiguity, geographic distance, and linguistic similarity were not reflected in similarity of gene frequencies. The gene frequenices found for Athabaskans appear to be the result of random processes—survivor effects and genetic drift of small isolated groups. They appear to be of no value in detecting ethnic relationships.

Spores of Bacillus subtilis NCTC 10073 were produced using three different methods on solid and liquid defined media. Thermal resistance at 90dC was examined in relation to the effects of cleaning procedures such as number of washings, speed and temperature of centrifugation on the spores. Ion-exchange spore forms (hydrogen and calcium) and coat-defective forms (urea-mercaptoethanol and urea-dithiothreitol-sodium dodecyl sulphate) were prepared from spores produced by the different spore production methods and compared for thermal resistance. The resistance patterns observed are discussed in terms of the spore properties of germinability. colony formation, dipicolinic acid and calcium contents.

Gorman, S.P. Scott, E.M. Hutchinson, E.P. 1984. Interaction of the Bacillus subtilis spore protoplast, cortex, ion-exchange and coatless forms with glutaraldehyde. Journal of Applied Bacteriology56, 95–102. Bacillus subtilis spores with altered ionic content were tested for their susceptibility to lysis with lysozyme or sodium nitrite following treatment with glutaraldehyde. The Ca-form was more sensitive to glutaraldehyde (pH 4.0.and pH 7.9) than the untreated or H-form. Removal of spore coat dramatically increased sensitivity of the spore to glutaraldehyde. Pretreatment of spores, the coats of which had been extensively removed, with glutaraldehyde (pH 7.9) reduced the rate of lysis by lysozyme and by sodium nitrite, whereas glutaraldehyde at pH 4.0.had little effect. Glutaraldehyde pretreatment (pH 4.0 and pH 7.9) reduced the amount of hexosamine released by lysozyme but not by nitrite from isolated cortical fragments. Spore protoplasts were more susceptible to 0.01% (w/v) glutaraldehyde at pH 4.0 and isolated spore coats adsorbed alkaline glutaraldehyde more rapidly. These results are discussed in terms of a possible mode of action of glutaraldehyde on the bacterial spore.

Estimation of the palaeo-dose and its associated error in TL dating requires assumptions to be made concerning the error structures of both the TL signal and dose. In this paper, we consider the sensitivity of palaeo-dose estimation to different error structures and describe techniques for estimation of the resultant palaeo-dose errors. We also indicate how the validity of the assumptions may be verified. We have taken the approach that procedures for the analysis of a glow curve at a single temperature must first be proved prior to their inclusion in a full analysis of the glow curves over a series of temperatures. Thus, the paper deals mainly with the analysis of the univariate structure of glow curves, i.e. estimation of the palaeo-dose and error at each temperature, but in the final section of the paper, we discuss briefly the multivariate structure of the results including automatic detection of plateau regions.

The resistance of spores of Bacillus subtilis NCTC 10073 to glutaraldehyde, sodium hypochlorite and povidone-iodine was compared. Revival of treated spores was examined by use of defined germination media and conditions, protein denaturing agents, ultrasonics and heat. Revival, obtained after treatment with each of the three chemical agents, originated under different sets of conditions and was of two recognizably distinct types. The results, including the evidence of electron microscopy, are discussed in terms of chemical-spore reactivity and the implications on their use and suitability as chemical sterilizers.

The resistance of spores of Bacillus subtilis NCTC 10073 to sodium hypochlorite and sodium hypochlorite-methanol combinations was compared with reference to concentration and pH. Activity against ion-exchange spores (H- and Ca-form) and coat-defective spores (SLS-, UME-, UMS-, UDT- and UDS-treated) was examined in relation to degree of coat protein solubilization. Methanol greatly enhanced hypochlorite activity at all stages of coat extraction. The degree of spore penetration and site of action is discussed.

An investigation of a possible relationship between Diphyllobothrium (fish tapeworm) infection and microcytic anaemia among Eskimos was carried out during 1957–1958 at the village of Hooper Bay in western Alaska. Such infections did not contribute to anaemia in residents of this region, a finding in agreement with those of Totterman 1947 in Finland. Species of diphyllobothriid cestodes recorded from the lower Kuskokwim River region are listed, and other helminths recorded from man in this area are briefly discussed. Schistocephalus solidus (Mueller 1776) is reported from man, apparently for the first time.

Increased bactericidal activity was observed on addition of inorganic cations to both acid and alkaline glutaraldehyde solutions. Mg2+ ion in particular potentiated activity and this was attributed to cell surface effects due to osmotic forces resulting in greater uptake of disinfectant, reduction in cell volume and interference with enzyme activity associated with the cell wall.

Spores of Bacillus subtilis NCTC 10073 were examined for susceptibility to two proprietary brands of povidone-iodine: an aqueous solution, Betadine and an alcoholic solution, Videne. Spores were converted to ion-exchange (Ca, H) and coat-defective (SLS-, UME-, UMS-, UDT- and UDS-treated) forms. The resistance of these to povidone-iodine was compared and related to uptake. Effects on spore protoplasts and cortex in relation to hexosamine release were also examined. The degree of spore penetration and site of action of povidone-iodine is discussed.