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PubMed2001

Effect of a polyhexamethylene biguanide mouthrinse on bacterial counts and plaque.

OBJECTIVES: For various clinical applications, polyhexamethylene biguanide (PHMB) has been used for many years as an antiseptic in medicine. Little is known, however, of its antibacterial activity in the oral cavity and its ability to inhibit plaque formation. In this study, a 0.04% PHMB mouthrinse (A) was compared with a negative control placebo rinse (10% ethanol, flavour) (B) and 2 positive control chlorhexidine rinses, one of which was a 0.12% aqueous solution (C) and the other a commercially available mouthrinse (Skinsept mucosa) diluted to a chlorhexidine concentration of 0.12% (D). MATERIAL AND METHODS: The study was a double-blind, randomised 4-replicate 4 x 4 Latin square cross-over design. Plaque regrowth was assessed with the Turesky et al. modification of the Quigley and Hein plaque index. The in vivo antibacterial effect was assessed by taking bacterial counts from the tooth surface (smears from the lingual surface of 16) and mucosa (smears from the buccal mucosa) 4 h after the 1st rinse with the preparations on day 1 and prior to the clinical examination on day 5. 16 volunteers participated, and on day 1 of each study period were rendered plaque-free, ceased toothcleaning, and rinsed 2x daily with the allocated mouthrinse. On day 5, plaque was scored and smears were collected according to the protocol. Washout periods were 9 days. Data were analysed using ANOVA with Tukey HSD adjustment for multiple comparisons (significance level alpha=0.05). RESULTS: Mouthrinses A, C, and D were significantly more effective in inhibiting plaque than the placebo (B). Mouthrinse C was significantly better than mouthrinses A and D, while mouthrinses D and A were equally effective in inhibiting plaque. Bacterial count reductions on the tooth surface with mouthrinse C were significantly greater compared to mouthrinse A and the placebo (B). The reduction of bacterial counts on the mucosa with C was significantly greater than with A and B after 4 h and significantly greater than with A, B and D after 5 days. Mouthrinse A reduced bacteria on the mucosa significantly more effectively than the placebo (B) after 4 h and 5 days, while mouthrinse D was more effective than the placebo (B) after 4 h. CONCLUSION: The results indicate that a 0.04% PHMB mouthwash inhibits plaque regrowth and reduces oral bacterial counts, and may be used in preventive applications in the oral cavity.

Adult↗
PubMed2005

[Residual leukocyte counting in plasma by a real-time genomic amplification assay].

BACKGROUND: Systematic plasma leukoreduction, which was introduced in France in April 1st 2001, has given rise to more sensitive methods for residual leukocytes counting. The technologies in application at this moment (Nageotte hemocytometers and flow cytometry methods) have been modified by a thirty fold sample concentration prior to analysis, inducing frequent downgrading of the plasma unit. So, in order to improve the detection threshold, we developed a more sensitive assay using "Real-Time Polymerase Chain Reaction" technology. MATERIALS AND METHODS: Real-time polymerase chain reaction was performed on a highly conserved HLADQalpha1 gene sequence. In order to determine the analytical performances of the method (accuracy, sensitivity, linearity and specificity) serial dilutions series ranging from 10(4) to 1 cells/ml were performed. A total of 18 series were prepared from three leukocyte stock solutions, by 1 in 10 serial dilutions in three plasmas completely devoided of white cells (called negative plasmas). To examine the specificity of the assay two negative controls were analyzed in each run. RESULTS: a sensitivity of 10 cells/ml (10(4) leukocytes/l) was achieved and the assay was linear between 10 and 10(4) cells/ml. The slope (-3.72) of the average standard curve calculated from all series, showed an amplification yield of 92.85%. CONCLUSION: we developed a quantitative assay for residual leukocytes in leukodepleted plasma, that agreed with the quality control requirement specifications.

Benzothiazoles↗
PubMed2006

The impact of daily Duddingtonia flagrans application to lactating ewes on gastrointestinal nematodes infections in their lambs in the Netherlands.

Two experiments were performed in 2002 and 2003 to evaluate the effect of biological control of gastrointestinal nematodes in sheep through the daily feeding of 500,000 chlamydospores of Duddingtonia flagrans/kg bodyweight to lactating ewes during the first 9 weeks with their young lambs on pasture. In both experiments four groups of eight ewes and their April-borne lambs were used. They were turned out on four separate plots (plots A) at the beginning of May, moved to similar separate plots after 3 (plots B) and 6 weeks (plots C), respectively, and weaning occurred after 9 weeks. In both experiments, two groups were fed spores daily while the two other groups served as controls. The effect of D. flagrans application was evaluated through faecal egg counts of ewes and lambs, the yield of faecal cultures in ewes, pasture larval counts and worm counts of lambs and tracer lambs. The results demonstrated no effect of D. flagrans application during the first 5 (2002) or 4 (2003) weeks. Subsequently, fungus application strongly reduced the yield in faecal cultures of the ewes. This was, however, not reflected in the pasture larval counts, but lower worm burdens were observed in tracer lambs of 'treated' plots C in 2002 than on those of 'control' plots. In 2003 worm burdens in 'treated' lambs returned to plots B were lower than those of 'control' lambs and a tendency for the same was observed for plots C. However, in all groups, lambs and tracer lambs developed severe haemonchosis.

Animals↗
PubMedDate not supplied

Investigation of diagnostic importance of platelet closure times measured by Platelet Function Analyzer--PFA 100 in dogs with endotoxemia.

This study was performed to evaluate the diagnostic importance of the platelet closure times measured by the Platelet Function Analyzer (PFA-100) in dogs with endotoxemia. E. coli endotoxin was given intravenously once, at the dose of 0.02 mg/kg or 1 mg/kg in groups I (n=9) and II (n=8), respectively. Normal saline (0.1 ml/kg) was injected in group III (n=8). The dogs were monitored for 48 h, and venous blood samples were collected prior to (baseline) and at intervals of 0.5, 1, 2, 4, 6, 8, 12, 24 and 48 h subsequent to the treatments. The white blood cell (WBC), platelet counts, and hematocrit (Hct) values were recorded. Platelet closure times were determined, using collagen/epinephrine (CEPI) and collagen/adenosine diphosphate (CADP) cartridges. Within 0.5 h after the endotoxin application baseline WBC and platelet counts (mean +/-SD) decreased significantly (p<0.001) to 2000 +/- 500 and 1850 +/- 200 cells/microl or 69.000 +/- 12.500 and 27.000 +/- 6.400 cells/microl in groups I and II, respectively. Platelet counts remained low during the first 1-48 h, but the WBC count was high at the 8th-48th h, in groups I and II, compared with baselines (p<0.001). After the application of the endotoxin, Hct values increased from baseline values of 37 +/- 3 or 39 +/- 2% to 48 +/- 2 or 51 +/- 3%, within 1 h (p<0.001), in groups I and II, respectively. Hct values in group II were notably higher (p<0.001) than those of group I, during the 2nd-48th h. Hematological parameters and closure times did not differ significantly throughout the study in group III. Baseline closure time ranged from 79 +/- 5 seconds (s) to 86 +/- 5 s for CADP and 144 +/- 13 s to 159 +/- 14 s for CEPI in all dogs (n=25). At 0.5 h after the endotoxin, the closure times of CADP as well as CEPI declined to 62 +/- 6 s and 76 +/- 8 s in group I (p<0.001) and 57 +/- 5 s and 75 +/- 6 s in group II (p<0.001). Afterwards, closure time prolonged to the levels of 280 +/- 8 s (CADP) and 294 +/- 5 s (CEPI) by 48 h (p<0.001) in group II, but returned to the baseline limit in group I. In conclusion, our results show that the shortened closure times may serve as a very early diagnostic sign of endotoxemia, prolonged closure times however may be used as an index for the severity of endotoxemia.

Animals↗
PubMed1988

An improved procedure for background correction in autoradiography.

In the event of weak autoradiographic labelling, the proportion of truly labelled cells or structures can be calculated from the frequency distributions of grains per area or cell structure for i = 0, 1,..., n grains using the results obtained for an experimental group after the application of a radioactively labelled substance and those obtained for a control group without radioactivity. The principle of this computer-aided method is also applicable when the grain counts are related to varying areas in histological sections.

Animals↗
PubMed1981

Trypan blue dye uptake and lactate dehydrogenase in adult rat hepatocytes--freshly isolated cells, cell suspensions, and primary monolayer cultures.

Leakage of lactate dehydrogenase and staining by the vital dye trypan blue were investigated in adult rat hepatocytes at the time of isolation, in suspensions up to 3 h and in primary monolayer cultures up to 3 d. These two parameters of plasma membrane integrity were found to correlate closely in hepatocyte suspensions, but to a lesser degree in monolayer cultures. Functional activity was demonstrated in culture by glucose consumption and lactic acid production. There was a balance of total lactate dehydrogenase (LDH) activity over time for both hepatocyte suspensions and cultures. Loss of LDH activity in the cell fraction was accompanied by a corresponding increase in enzyme activity in the media fraction. Lactate dehydrogenase activity per dye-excluding hepatocyte was calculated to be 9.2 +/- 1.5 X 10(-6) IU assayed at 37 degrees C for 25 preparations of isolated hepatocytes. The results suggest that leakage of cytoplasmic enzyme and vital dye staining are of comparable sensitivity in evaluating hepatocyte preparations. Measurement of LDH leakage offers a less subjective alternative to cell counting procedures and is applicable to both attached and suspended cells.

Animals↗
PubMed2000

Comparative evaluation of scatter correction techniques in 3D positron emission tomography.

Much research and development has been concentrated on the scatter compensation required for quantitative 3D positron emission tomography (PET). Increasingly sophisticated scatter correction procedures are under investigation, particularly those based on accurate scatter models and iterative reconstruction-based scatter compensation approaches. The main difference among the correction methods is the way in which the scatter component in the selected energy window is estimated. Monte Carlo methods provide further insight and might in themselves offer a possible correction procedure. Five scatter correction methods were compared in this study where applicable: the dual-energy window (DEW) technique, the convolution-subtraction (CVS) method, two variants of the Monte Carlo-based scatter correction technique (MCBSCI and MCBSC2) and our newly developed statistical reconstruction-based scatter correction (SRBSC) method. These scatter correction techniques were evaluated using Monte Carlo simulation studies, experimental phantom measurements and clinical studies. Accurate Monte Carlo modelling is still the gold standard since it allows the separation of scattered and unscattered events and comparison of the estimated and true unscattered component. In this study, our modified version of Monte Carlo-based scatter correction (MCBSC2) provided a good contrast recovery on the simulated Utah phantom, while the DEW method was found to be clearly superior for the experimental phantom studies in terms of quantitative accuracy at the expense of a significant deterioration in the signal-to-noise ratio. On the other hand, the immunity to noise in emission data of statistical reconstruction-based scatter correction methods makes them particularly applicable to low-count emission studies. All scatter correction methods gave very good activity recovery values for the simulated 3D Hoffman brain phantom, which averaged within 3%. The CVS and MCBSC 1 techniques tended to overcorrect while SRBSC undercorrected for scatter in most regions of this phantom. It was concluded that all correction methods significantly improve the image quality and contrast compared to the case where no correction is applied. Generally, it was shown that the differences in the estimated scatter distributions did not have a significant impact on the final quantitative results. The DEW method showed the best compromise between ease of implementation and quantitative accuracy, but entailed a significant deterioration in the signal-to-noise ratio.

Algorithms↗
PubMed1994

Computed detection and quantitative morphometry of Alzheimer senile plaques.

Senile plaques (SP) are the most characteristic neuropathologic lesions of Alzheimer's disease (AD) and studies of plaque cortical distribution, density, and morphology may lead to new information about the origin and pathogenesis of this disease. We have developed an automated computer image analysis program to detect SP (including diffuse and mature forms) and to measure SP size, shape, and fractional area or load in digital micrographs of silver-stained tissue sections. The plaques are detected with adaptive thresholding, requiring no user interaction. Measures of SP size, morphology, and load are readily calculated from the pixel values in the detected SP features. These measurements are achieved accurately and exhaustively, and this method offers an alternative to manual SP counting. We demonstrate its application to 4 cases spanning the full range of the severity of the disease.

Adult↗
PubMed2005

Screening for physical activity in family practice: evaluation of two brief assessment tools.

BACKGROUND: Physical activity (PA) is relevant to the prevention and management of many health conditions in family practice. There is a need for an efficient, reliable, and valid assessment tool to identify patients in need of PA interventions. METHODS: Twenty-eight family physicians in three Australian cities assessed the PA of their adult patients during 2004 using either a two- (2Q) or three-question (3Q) assessment. This was administered again approximately 3 days later to evaluate test-retest reliability. Concurrent validity was evaluated by measuring agreement with the Active Australia Questionnaire, and criterion validity by comparison with 7-day Computer Science Applications, Inc. (CSA) accelerometer counts. RESULTS: A total of 509 patients participated, with 428 (84%) completing a repeat assessment, and 415 (82%) accelerometer monitoring. The brief assessments had moderate test-retest reliability (2Q k=58.0%, 95% confidence interval [CI]=47.2-68.8%; 3Q k=55.6%, 95% CI=43.8-67.4%); fair to moderate concurrent validity (2Q k=46.7%, 95% CI=35.6-57.9%; 3Q k=38.7%, 95% CI=26.4-51.1%); and poor to fair criterion validity (2Q k=18.2%, 95% CI=3.9-32.6%; 3Q k=24.3%, 95% CI=11.6-36.9%) for identifying patients as sufficiently active. A four-level scale of PA derived from the PA assessments was significantly correlated with accelerometer minutes (2Q rho=0.39, 95% CI=0.28-0.49; 3Q rho=0.31, 95% CI=0.18-0.43). Physicians reported that the assessments took 1 to 2 minutes to complete. CONCLUSIONS: Both PA assessments were feasible to use in family practice, and were suitable for identifying the least active patients. The 2Q assessment was preferred by clinicians and may be most appropriate for dissemination.

Adolescent↗
PubMed2004

A new concept for ultrasensitive fluorescence measurements of molecules in solution and membrane: 1. Theory and a first application.

Just because there is an average of one molecule in the observation volume of a solution or membrane (single-phase), one cannot say that this is an individual molecule since many different single molecules measured one by one or the same single, individual molecule not leaving the detection volume on time average can cause a single-molecule event. The latter case is of interest and allows the continuous observation of one and the same single molecule without averaging over many 'different' single molecules. For the first time a universal theoretical and experimental framework is presented for the continuous observation of the same single, individual molecule without immobilization, hydrodynamic flow, or burst size histograms of fluorescence intensity traces. In this original article, the stochastic approach is derived and its main characteristics are demonstrated with the free fluorophore rhodamine-green in solution for simpler experimental realization. Single (solution)-phase single-molecule fluorescence auto- (or two-color cross-) correlation spectroscopy (SPSM-FCS) is used as a specific application in order to count the absolute number of molecules in the observation volume. The absolute number of molecules, the diffusion coefficient of the single fluorescent molecule, the lower limit of distance, and the molar concentration of the bulk phase (solution) were directly obtained from the measured auto- or (cross)-correlation curves of the SPSM-FCS experiments. For this purpose, the detection volume that was measured was less then 1 fl (10(-15) l). Then, a concentration of the bulk solution was chosen in such a way that the probability of detecting more than one molecule in the detection volume was very small. The Poisson probability was experimentally determined for the absolute number of molecules depending upon a specified bulk concentration. From the diffusion coefficient of the molecule, it was found that the probability of the molecule diffusing out of the probe volume during the measurements was negligibly small.

Fluorescent Dyes↗
PubMed2002

Base pair mismatch recognition using plasmon resonant particle labels.

We demonstrate the use of silver plasmon resonant particles (PRPs), as reporter labels, in a microarray-based DNA hybridization assay in which we screen for a known polymorphic site in the breast cancer gene BRCA1. PRPs (40-100 nm in diameter) image as diffraction-limited points of colored light in a standard microscope equipped with dark-field illumination, and can be individually identified and discriminated against background scatter. Rather than overall intensity, the number of PRPs counted in a CCD image by a software algorithm serves as the signal in these assays. In a typical PRP hybridization assay, we achieve a detection sensitivity that is approximately 60 x greater than that achieved by using fluorescent labels. We conclude that single particle counting is robust, generally applicable to a wide variety of assay platforms, and can be integrated into low-cost and quantitative detection systems for single nucleotide polymorphism analysis.

Base Pair Mismatch↗
PubMed2003

Subeschar clysis in deep burns.

Six hundred thirteen patients with deep burn of up to 50% total body surface area (TBSA) were treated with 0.25% povidone iodine subeschar clysis (PVP-SEC) in addition to surface application of povidone iodine + Neosporin in the form of "crust". The results were compared with those of 595 age, sex and percentage of burn, matched patients treated only by "crust application". The quantitative bacterial count showed significantly less incidence of infection on the 7th and 8th days post treatment (P<0.001). The organisms identified were predominately Staphylcocous aureus and Pseudomonas aeroginosa. Significantly more number of patients, with burns up to 50% TBSA, could be grafted within 20 days in the SEC group. The graft acceptance rate in this group was 90%.

Adult↗
PubMedDate not supplied

Radioactivity standardization in South Africa.

South Africa's national radioactivity measurement standard is maintained at a satellite laboratory in Cape Town by the National Metrology Laboratory (NML) of the Council for Scientific and Industrial Research. Standardizations are undertaken by a number of direct methods utilizing liquid scintillation counting (LSC). The successful application of LSC to the 4pi beta-gamma coincidence method is reviewed. The activity unit is maintained through radionuclide specific calibration factors relating to a pressurized re-entrant well type ionization chamber. A comparison is made between normalized factors given by the manufacturer and deduced factors obtained by a method used to transfer calibration factors from the International Reference System of the International Bureau of Weights and Measures based on the NML's own absolute standardizations.

Journal Article↗
PubMed2000

A multimethodic and multiparametric system for standardisation of radionuclides

A measuring system with simultaneous application of four independent counting methods for standardization of solutions of different radionuclides is presented. The result obtained using four methods allows for mutual verification and gives the final result with a higher confidence level. The multidetection scintillation head for 4pi(LS)-gamma coincidence and anticoincidence with double-parametric registration, as well as the electronic diagram, is described. The four methods, comprised in the presented system, are described. Using the multimethodic and multiparametric system, massic activity of the solutions of the following radionuclides with various decay schemes has been measured: 133Ba, 90Sr, 99Mo, 85Sr, 65Zn, 139Ce, 192Ir, 204Tl, 60Co, 241Am, 152Eu, 169Yb. The results of the methods were analyzed using the figure of merit evaluated following the ISO/IEC guide 47-1. The system seems to allow the analysis of the correctness of the extrapolation curves, activity vs. the LSC efficiency, applied in 4pi(LS)-gamma methods.

Journal Article↗
PubMed2006

Application of capture-recapture models to estimation of protein count in MudPIT experiments.

MudPIT is an automated shotgun proteomics approach that enhances the separation of peptides for sequencing by mass spectrometry analysis.We here adapt a mathematical model from ecology, namely, the capture-recapture model with a closed population and time-varying and heterogeneous individual probabilities of capture, to model the number of peptide identifications across the various cycles of a typical MudPIT experiment. In the absence of any prior information on abundance levels, the model can be used to estimate the total number of proteins in the experimental sample. We apply the model to a recent MudPIT-based experiment to estimate the total number of rat lung endothelial cell surface proteins. The model provides some practical guidelines for planning MudPIT experiments.

Animals↗
PubMed1995

Exceptional motifs in different Markov chain models for a statistical analysis of DNA sequences.

Identifying exceptional motifs is often used for extracting information from long DNA sequences. The two difficulties of the method are the choice of the model that defines the expected frequencies of words and the approximation of the variance of the difference T(W) between the number of occurrences of a word W and its estimation. We consider here different Markov chain models, either with stationary or periodic transition probabilities. We estimate the variance of the difference T(W) by the conditional variance of the number of occurrences of W given the oligonucleotides counts that define the model. Two applications show how to use asymptotically standard normal statistics associated with the counts to describe a given sequence in terms of its outlying words. Sequences of Escherichia coli and of Bacillus subtilis are compared with respect to their exceptional tri- and tetranucleotides. For both bacteria, exceptional 3-words are mainly found in the coding frame. E. coli palindrome counts are analyzed in different models, showing that many overabundant words are one-letter mutations of avoided palindromes.

Bacillus subtilis↗
PubMed1982

Determination of the number of superhelical turns by the hyperchromicity of partially denatured covalently-closed DNA molecules.

A novel method of determining the number of superhelical turns of a covalently-closed plasmid DNA is described. It relies on the determination of the hyperchromicity, and hence the proportion of unstacked basepairs, of a partially heat-denatured sample which co-migrates during electrophoresis with nicked circular duplex DNA. The values obtained for plasmid pBR beta G DNA at 4 degrees C (-29.8 and -33.5 in the two buffers used) agree closely with the values obtained in parallel by topoisomer band-counting. Our method is less precise than band-counting but is readily applicable to determining the superhelicity of very large DNA molecules. Our results confirm earlier findings that magnesium-containing buffers cause an increase in the duplex winding angle, and hence an increase in the number of negative superhelical turns.

Animals↗
PubMedDate not supplied

Microbial contamination in allografted wound beds in patients with burns.

Microbial wound contamination has been recognized as a cause of autograft skin graft failure. Human cadaver allograft is believed to decrease or control microbial wound contamination, and by its adherence to the wound bed, to indicate a sufficiently low microbial count to allow successful application of autograft. Suboptimal "take" of more fragile cultured skin grafts after removal of adherent allograft led us to reexamine the microbial population of these wound beds. Immediately before autografts were placed, wound beds beneath adherent allograft were biopsied for quantitative microbiologic analysis. Eighty tissue biopsy specimens from 21 patients were examined. No patients had signs of infection or sepsis at the time of biopsy. Fifty-seven percent of all patients had positive cultures beneath adherent allograft skin. Twenty-one percent of all cultures revealed "infected" wound beds (greater than 100,000 colonies per gram), and 30% of all wounds were "colonized" (less than 100,000 colonies per gram). These data suggest that take and vascularization of allograft does not guarantee that a wound bed is free of microbial contamination.

Adolescent↗