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At least 505 records · Page 28Linked to original sources

Measuring tissue oxygen tension: a review.

Because of technological advances in tissue oximetry, clinicians and scientists have a better understanding of the role of oxygen in wound healing. In wound care and hyperbaric medicine applications, an oximeter is principally used with vascular assessment to help determine amputation level and to estimate healing potential. With the current emphasis on cost savings in the managed care setting, transcutaneous oximetry (PtcO2) has gained importance as a tool for predicting potential candidates for hyperbaric oxygen (HBO2) therapy. It is used to identify the presence of hypoxia in wounded tissue, to predict the responders to hyperoxia and in some instances to determine when HBO2 treatment is complete. This literature review describes the principal current methods for measuring tissue O2 and the values obtained in normal and wounded tissue under both normobaric and hyperbaric conditions. The review includes the Jefferson C. Davis Wound Care and Hyperbaric Medicine Center protocol for PtcO2 assessment of potential HBO2 candidates and suggestions for obtaining reproducible PtcO2 data.

Blood Gas Monitoring, Transcutaneous↗

Ca2+ buffering in the heart: Ca2+ binding to and activation of cardiac myofibrils.

The measurement of cardiac Ca(2+) transients using spectroscopic Ca(2+) indicators is significantly affected by the buffering properties of the indicators. The aim of the present study was to construct a model of cardiac Ca(2+) buffering that satisfied the kinetic constraints imposed by the maximum attainable rates of cardiac contraction and relaxation on the Ca(2+) dissociation rate constants and which would account for the observed effects of (19)F-NMR indicators on the cardiac Ca(2+) transient in the Langendorff-perfused ferret heart. It is generally assumed that the Ca(2+) dependency of myofibril activation in cardiac myocytes is mediated by a single Ca(2+)-binding site on troponin C. A model based on 1:1 Ca(2+) binding to the myofilaments, however, was unable to reproduce our experimental data, but a model in which we assumed ATP-dependent co-operative Ca(2+) binding to the myofilaments was able to reproduce these data. This model was used to calculate the concentration and dissociation constant of the ATP-independent myofilament Ca(2+) binding, giving 58 and 2.0 microM respectively. In addition to reproducing our experimental data on the concentration of free Ca(2+) ions in the cytoplasm ([Ca(2+)](i)), the resulting Ca(2+) and ATP affinities given by fitting of the model also provided good predictions of the Ca(2+) dependence of the myofibrillar ATPase activity measured under in vitro conditions. Solutions to the model also indicate that the Ca(2+) mobilized during each beat remains unchanged in the presence of the additional buffering load from Ca(2+) indicators. The new model was used to estimate the extent of perturbation of the Ca(2+) transient caused by different concentrations of indicators. As little as 10 microM of a Ca(2+) indicator with a dissociation constant of 200 nM will cause a 20% reduction in peak-systolic [Ca(2+)](i) and 30 microM will cause approx. 50% reduction in the peak-systolic [Ca(2+)](i) in a heart paced at 1.0 Hz.

Adenosine Triphosphate↗

Utility of the hot-wire spirometer.

A comparative study has been carried out to investigate the performance of the hot-wire spirometer and the Rollingseal spirometer; the reproducibility of the data generated by an individual examiner; and the validity of data obtained by an "inexperienced examiner" using the hot-wire spirometer. The following results were obtained: 1. The experimental data given by the hot-wire spirometer were about 5-6% higher for FVC and FEV1, and about 10-12% higher for PFR, V50, and V25, respectively, when compared to the data generated by the Rollingseal spirometer. 2. The reproducibility of the data produced by the hot-wire spirometer operated by an "experienced examiner" was good, as the percent difference was about +/- 3%. 3. Reliable data were obtained with the spirometer even by an "inexperienced examiner" if he/she has gone through an on-site training concerning instrumentation and measurement methods for spirography.

Evaluation Studies as Topic↗

Interlaboratory validation of a CD71-based flow cytometric method (Microflow) for the scoring of micronucleated reticulocytes in mouse peripheral blood.

An interlaboratory study was performed to validate an anti-CD71/flow cytometry-based technique for enumerating micronucleated reticulocytes (MN-RETs) in mouse peripheral blood. These experiments were designed to address International Workshop on Genotoxicity Test Procedures validation criteria by evaluating the degree of correspondence between MN-RET measurements generated by flow cytometry (FCM) with those obtained using traditional microscopy-based methods. In addition to these cross-methods data, flow cytometric MN-RET measurements for each blood sample were performed at two separate sites in order to evaluate the reproducibility of data between laboratories. In these studies, groups of male CD-1 mice were treated with vehicle (saline or vegetable oil), a negative control (saline or vegetable oil), or four dose levels of five known genotoxicants (clastogens: cyclophosphamide, benzo[a]pyrene, 5-fluorouracil, methotrexate; aneugen: vincristine sulfate). Exposure occurred on 3 consecutive days via intraperitoneal injection, and blood samples were obtained approximately 24 hr after the final treatment. MN-RET frequencies were determined for each sample based on the analysis of 2,000 (microscopy) and 20,000 (FCM) reticulocytes. Regardless of the method utilized, each genotoxic agent was observed to cause statistically significant increases in the frequency of MN-RETs, and each response occurred in a dose-dependent manner. Spearman's correlation coefficient (rs) for FCM versus microscopy-based MN-RET measurements (nine experiments, 252 paired measurements) was 0.740, indicating a high degree of correspondence between methods. The rs value for all flow cytometric MN-RET measurements performed at the two independent sites was 0.857 (n = 248), suggesting that the automated method is highly transferable between laboratories. Additionally, the flow cytometric system offered advantages relative to microscopy-based scoring, including a greater number of cells analyzed, much faster analysis times, and a greater degree of objectivity. Collectively, data presented in this report suggest that the overall performance of mouse peripheral blood micronucleus tests is enhanced by the use of the flow cytometric scoring procedure.

Animals↗

International validation study for the determination of chloramphenicol in bovine muscle.

A gas chromatographic (GC) procedure for the quantitation and GC/negative ion chemical ionization mass spectrometric (NICIMS) confirmation of chloramphenicol in calf muscle tissue was the subject of a validation study. Five analysts representing 5 laboratories in 4 countries participated in the quantitative method and analyzed 7 randomly numbered blind triplicates at 4 fortified and 3 incurred tissue concentrations on 3 separate days. The chloramphenicol concentrations ranged from 0 to 2.5 ppb. All data were reported to 3 significant figures. The coefficients of variation were 9.5-28.7% for repeatability and 14.6-38% over the study range for reproducibility. NICIMS data representing 3 laboratories in 3 countries successfully confirmed chloramphenicol in samples at 0.6 ppb or greater with no false positives in blank tissues.

Animals↗

Longitudinal study of intrahepatic portal flow: a novel approach using Doppler technique.

The present study investigates whether the application of the summation of portal blood flow (SPBF), measured from the left umbilical portion and the right anterior branch by Doppler, could be of use in the longitudinal study of intrahepatic portal flow or not. Forty-five subjects were divided into two groups. Group I subjects included males and post-menopausal females. Thirty-seven subjects (19 normal volunteers, 12 subjects with chronic hepatitis, six cirrhotics) were investigated by the more experienced doctor A at 9 day intervals for an intra-observer study. Among them, seven subjects (four normal volunteers, three with chronic hepatitis) were also investigated consecutively at 10 min apart by doctor B on days 1 and 10 for an interobserver study. Group II subjects included eight normal subjects with a regular menstrual cycle. They were investigated by doctor A during menstruation, mid-follicular, ovulatory and mid-luteal phases of the menstrual cycle to evaluate the possible impact of the menstrual cycle on SPBF measurement. The reproducibility of measurements was assessed by the coefficient of variation (CV) and the reproducibility coefficient. Data are expressed as mean +/- s.d. The CV of the SPBF for intra-observer and interobserver study were 4.88 +/- 3.36 and 4.19 +/- 2.53, respectively. All differences of SPBF were within their corresponding reproducibility limits. The results of SPBF measurements obtained from different phases of the menstrual cycle were variable. Only the comparison between the data obtained from mid-follicular and mid-luteal phases could demonstrate acceptable CV values (3.98 +/- 3.12%) and all the differences in SPBF measurements were within the repeatability limit. In conclusion, Doppler SPBF measurement may be useful in the longitudinal evaluation of intrahepatic portal flow changes.

Adult↗

[Reproducibility of measuring retinal nerve fiber density. Comparison of optical coherence tomography with the nerve fiber analyzer and the Heidelberg retinal tomography device].

BACKGROUND: Glaucoma is associated with a progressive loss of retinal ganglion cell axons and the evaluation of the thickness of the retinal nerve fibre layer (RNFL) is important for early diagnosis and follow-up. Results of RNFL thickness measurements may vary according to the devices used. Therefore, the present study has analysed and compared the reproducibility of data obtained with three different instruments. METHODS: RNFL thickness was determined using a circular identification line around the optic nerve head by optical coherence tomography (OCT), a Heidelberg Retina Tomograph (HRT) and a Nerve Fiber Analyser (NFA). Each eye was tested six times. The values for RNFL thickness were calculated for the superior, inferior, nasal and temporal areas. A one-way analysis of variance with random effects was chosen to estimate the variance components. The intraclass correlation coefficient (ICC) as a measure of reliability and the coefficient of variation (CV%) as a measure of precision were determined to estimate the reproducibility of RNFL thickness measurements. RESULTS: With all three instruments, the reproducibility of measurements between the superior, inferior, temporal and nasal areas did not differ significantly suggesting that reproducibility does not depend on the thickness of the RNFL. Data obtained by NFA showed the highest ICC values (0.9) and lowest CV% values (7.0), followed by those obtained by HRT (reliability ICC 0.8; precision CV% 12). A markedly lower degree of reproducibility was found for the OCT compared to NFA and HRT (reliability ICC 0.5; precision CV% 15). CONCLUSION: The comparison of RNFL thickness measurements with OCT, NFA, and HRT showed that the NFA yielded the most reliable results followed by HRT.

Adolescent↗

Reproducibility of swallowing over time in healthy elderly adults.

The purpose of this study was to evaluate the reproducibility of aspects of swallowing assessed over time by the Exeter Dysphagia Assessment Technique. Fourteen healthy elderly adults were assessed during the morning on four consecutive days. During day 1, the assessment was repeated ten times. During days 2, 3, and 4 the assessment was carried out once and at the same time of the day. This allowed the reproducibility of the data to be examined between sessions and between days. The results show that the reproducibility of data between sessions was moderate to very good for all of the items evaluated (median kappas = 0.56-0.82, ICCs = 0.7-0.8). Between-day reproducibility was also moderate to very good for nine of the variables measured (median kappas = 0.58-0.96, ICCs = 0.53-0.84) and poor for one (ICC = 0.15). We conclude that the majority of variables examined by the Exeter Dysphagia Assessment Technique are reliable when swallowing is assessed repeatedly and on different days.

Aged↗

A metabolic model describing the H2O2 elimination by mammalian cells including H2O2 permeation through cytoplasmic and peroxisomal membranes: comparison with experimental data.

We have constructed a metabolic model describing the H2O2 elimination by mammalian cells. It comprises three compartments (medium, cytosol, and peroxisome) separated by cytoplasmic and peroxisomal membranes, and H2O2 moves across the membranes with different permeation rate constants. Catalase localizes to peroxisomes, while glutathione peroxidase (GPx) and GSH recycling system (glutathione reductase (GR) and the oxidative pentose phosphate pathway (PPP)) localize to cytosol. The rates of individual enzyme reactions were computed using the experimentally determined activities and rate equations known for mammalian enzymes. Using the model, the concentration dependence of H2O2 elimination rate was obtained by numerical simulation and was compared with experimental data obtained previously with cultured mammalian cells (fibroblasts, human umbilical vein endothelial cells (HUVEC), and PC12 cells). The model was shown to be able to reproduce the data well by assuming appropriate values for the permeability rate constants. The H2O2 permeability coefficients thus estimated for cytoplasmic and peroxisomal membranes were in the same order of magnitude, except that the value for cytoplasmic membrane of PC12 cell was significantly smaller. The results suggest that the membrane permeability is one of the rate-limiting factors in the H2O2 elimination by mammalian cells. Using the model and estimated parameter values, we have examined the rate-limiting enzyme of the metabolic system, as well as the intracellular H2O2 concentration under steady-state and non-steady-state conditions.

Animals↗

Reproducibility of spinal back-contour measurements taken with raster stereography in adolescent idiopathic scoliosis.

200 children with a mean age of 12.7 years were measured with the Quantec Spinal Image System (QSIS), which uses computerized raster stereography technology. The aim of the study was to assess the intraobserver reproducibility of QSIS metrics in scoliosis patients and to quantify the effect of postural sway on the measurements. Children were randomly assigned to 1 of 2 groups: Group 1 contained 198 subjects, having 3 digitized measurements of 1 scan; Group 2 contained 200 subjects, undergoing 3 separate QSIS scans with 1 measurenent of each scan. Random-effects variance components models were fit to each outcome variable of interest (subject, scan or measurement) separately for the single scan dataset and the 3 scan dataset. They revealed that data from Group 2 contained greater reliability than data from Group 1 (reliability > or = 80%). The reliability of these parameters was perfect if one performed 3 scans and 3 measurements per scan. Results demonstrate that the QSIS with 3 scans and 1 measurement yields reproducible data from mild idiopathic scoliosis patients and that postural sway has minimal effect on data reproducibility.

Child↗

Carbohydrate metabolism in Dictyostelium discoideum: I. Model construction.

A model of the transitional changes which occur in fluxes and metabolite concentrations during the course of differentiation of Dictyostelium discoideum is constructed, based on extensive experimental data collected from D. discoideum. Input to the model includes metabolite compartmentation and fluxes, determined by radiotracer analyses, and enzyme mechanisms and constants. This transition model reproduced experimentally measured changes in metabolite concentrations over the last 15 hr of differentiation. The real test of a mathematical model is its ability to correctly predict the effects of an altered environment or a mutation. This model demonstrates that ability. Constraints on model parameters are also examined, and many of the altered parameters result in a model which is unable to reproduce experimental data. These results imply that the model represents many aspects of metabolism in vivo and will provide a means of examining rate-limiting events during differentiation and aging of this organism.

Animals↗

Rationalization of quantitative tooth surface loss data for epidemiological research.

Recent UK National Surveys have revealed a high prevalence of tooth surface loss (TSL) because of erosion in Children and Adolescents. Although digital surface mapping and surface matching techniques may be used to quantify its progression with time, reporting TSL of individuals as a function of either mean depth loss or volume loss, this can be inadequate when transferred for epidemiological analysis. For example, a tooth displaying multiple regions of depth loss may deserve to be distinguished from a tooth with a single localised area of erosion, although the total volume change may be the same. A potential solution, explored here, is to use both the quantitative wear data and colour coded surface representation plots generated by such methods to arrive at a single categorical wear score [1 (< or = 5% of surface exhibits TSL) to 5 (> or = 51% of surface exhibits TSL)]. Two examiners independently categorized, on two separate occasions, the TSL of 53 maxillary incisors (26 subjects observed at baseline and 9 months). Their performance was assessed for intra- and inter-examiner agreement by; (i) calculating the percentage of agreement, (ii) a Wilcoxon matched-pairs signed rank test. Thereafter, in cases of disagreement a consensus score was allocated and a regression analysis of this versus the numerically derived percentage of the surface that had undergone change was carried out. Intra- and inter-examiner agreement was 100 and 88.7% respectively and for the inter-examiner comparison P = 0.0456. The categorical scores and the quantitative wear data correlated linearly (R = 0.82). It is concluded that the method used to rationalize the quantitative TSL data is both reproducible and reflects the quantitative data. Its use should be helpful in epidemiological TSL studies but continued vigilance is required in cases of examiner disagreement.

Adolescent↗

Normal brain volume measurements using multispectral MRI segmentation.

The performance of a supervised k-nearest neighbor (kNN) classifier and a semisupervised fuzzy c-means (SFCM) clustering segmentation method are evaluated for reproducible measurement of the volumes of normal brain tissues and cerebrospinal fluid. The stability of the two segmentation methods is evaluated for (a) operator selection of training data, (b) reproducibility during repeat imaging sessions to determine any variations in the sensor performance over time, (c) variations in the measured volumes between different subjects, and (d) variability with different imaging parameters. The variations were found to be dependent on the type of measured tissue and the operator performing the segmentations. The variability during repeat imaging sessions for the SFCM method was < 3%. The absolute volumes of the brain matter and cerebrospinal fluid between subjects varied quite large, ranging from 9% to 13%. The intraobserver and interobserver reproducibility for SFCM were < 4% for the soft tissues and 6% for cerebrospinal fluid. The corresponding results for the kNN segmentation method were higher compared to the SFCM method.

Adult↗

Application of array CGH on archival formalin-fixed paraffin-embedded tissues including small numbers of microdissected cells.

Array-based comparative genomic hybridisation (aCGH) has diverse applications in cancer gene discovery and translational research. Currently, aCGH is performed primarily using high molecular weight DNA samples and its application to formalin-fixed and paraffin-embedded (FFPE) tissues remains to be established. To explore how aCGH can be reliably applied to archival FFPE tissues and whether it is possible to apply aCGH to small numbers of cells microdissected from FFPE tissue sections, we have systematically performed aCGH on 15 pairs of matched frozen and FFPE astrocytic tumour tissues using a well-established in-house human 1 Mb BAC/PAC genomic array. By spiking tumour DNA with normal DNA, we demonstrated that at least 70% of tumour DNA was required for reliable aCGH analysis. Using aCGH data from frozen tissue as a reference, it was found that only FFPE astrocytic tumour tissues that supported PCR amplification of >300 bp DNA fragment provided high quality, reproducible aCGH data. The presence of necrosis in a tissue specimen had an adverse effect on the quality of aCGH, while fixation in formalin for up to 96 h of fresh tissue did not appear to affect the quality of the result. As little as 10-20 ng DNA from frozen or FFPE tissues could be readily used for aCGH analysis following whole genome amplification (WGA). Furthermore, as few as 2000 microdissected cells from haematoxylin-stained slides of archival FFPE tissues could be successfully used for aCGH investigations when WGA was used. By careful assessment of DNA integrity and review of histology, to exclude necrosis and select specimens with a high proportion of tumour cells, it is feasible to preselect archival FFPE tissues adequate for aCGH analysis. With the help of microdissection and WGA, it is also possible to apply aCGH to histologically defined lesions, such as carcinoma in situ.

Astrocytoma↗

Initial statistical clustering: definition of temporary clusters of monoclonal antibodies for the duration of the workshop.

Statistical clustering was performed on flow cytometry data generated from the reactivities of 189 mAbs with 56 cell preparations or cell lines (targets). Six targets were excluded from the statistical clustering because of non-reproducibility of data from the internal controls. Thirty-six temporary clusters (TC) were formed which were subjected to further examination in the second phase of the workshop.

Animals↗

The Dutch EPIC food frequency questionnaire. I. Description of the questionnaire, and relative validity and reproducibility for food groups.

BACKGROUND: A self-administered food frequency questionnaire was developed for the Dutch cohort of the European Prospective Investigation into Cancer and Nutrition (EPIC). Habitual consumption of 178 food items can be calculated from the questionnaire data. METHODS: Reproducibility and relative validity for food group intake were investigated in a population of 121 Dutch men and Women. The questionnaire was administered three times at 6-month intervals in order to determine the reproducibility. To assess the relative validity 12 monthly 24-hour recalls served as the reference method. RESULTS: Spearman rank order correlation coefficients between estimates of food group intake assessed by repeated questionnaires ranged from 0.45 to 0.92. For men, Spearman correlation coefficients between estimates of food group intake based on the questionnaire and those based on 24-hour recalls ranged from 0.21 for cooked vegetables to 0.78 for sugar and sweet products, with a range of 0.61. For women the median was 0.53, with a minimum of 0.31 for vegetables and a maximum of 0.87 for alcoholic beverages. The photographs in the questionnaire for the estimation of portion sizes contributed little to the relative validity of the ranking of subjects. However, on the group level most median food group estimates based on photographic portion sizes were closer to the median intakes as assessed by 24-hour recalls than those based on standard portion sizes. CONCLUSIONS: The questionnaire seems adequate for ranking Dutch EPIC subjects according to intake of most food groups, although the relative validity for some food groups, such as vegetables and fish, remains of concern.

Adult↗

Evaluation of a 12-lead digital Holter system for 24-hour QT interval assessment.

BACKGROUND: Drug induced QT prolongation may precipitate life threatening cardiac arrhythmias. Evaluation of the QT prolonging effect of new pharmaceutical agents in a 'thorough QT/QTc study' is being mandated by FDA. The purpose of this study was to evaluate an automated 12-lead digital Holter system for a thorough QT/QTc study. METHODS: Five healthy volunteers underwent 24-hour digital Holter monitoring. Each recording underwent a fully automated QT analysis (AQA) followed by an onscreen complete manual over read (MOR). Each recording was analyzed twice at least 2 weeks apart. The effect of data sampling (5-min segment/hour), the system sensitivity to detect 5-ms increase in QT, and the ability to assess circadian variation were evaluated. RESULTS: The AQA resulted in identical QT for the first and second analyses, but with obvious errors in QT measurements. Compared to the complete onscreen MOR, the mean QT was longer with AQA (416 +/- 41 vs. 387 +/- 30 ms, p < 0.001), correlation; r = 0.3. The reproducibility of AQA with complete MOR was very good (QT: 387 +/- 30 vs. 387 +/- 30 ms, coefficient of variation: 0.2%, r = 0.986. The 5-min mean QT intervals correlated well with the hourly mean QT intervals (r = 0.994, p < 0.001, coefficient of variation = 1 ms) and both showed a similar circadian variation. The system was sensitive to detect a 5-ms change in QT intervals (5 +/- 2 ms, coefficient of variation = 0.6%, r = 0.998, p < 0.001). CONCLUSIONS: The AQA is not an acceptable method, while the automatic analysis with complete MOR is a highly sensitive and reproducible method. Data sampling by analyzing 5-min segments per hour is sensitive and reproducible.

Adult↗

Evaluation of qsars for ecotoxicity: a method for assigning quality and confidence.

Validation of a quantitative structure-activity relationship (QSAR) is now considered as an integral part of its development. Assessment of the quality of a QSAR and the confidence that may be placed in predictions from it are vital to any validation procedure. A number of terms associated with the quality of a QSAR, confidence in that QSAR, or both may be quantified. These terms include the: (1) goodness of fit of the model (r2); (2) predictivity of the model (Q2); (3) stability of the model described as the difference between fit and predictivity (Dfp); (4) number of compounds used in the training set (Nc); (5) number of descriptors used in the model (Nd); (6) range of toxicity values (Tr); (7) number of mechanisms of toxic action covered by the training set (Nm), as well as two factors associated with the biological data-confidence associated with, (8) reproducibility of the data (Rconf) and (9) confidence in the source of the data (Sconf). While all these factors may influence the quality of, and/or confidence in a particular QSAR, each varies within different limits. To enable a quantitative assessment of quality and confidence in a QSAR, the terms deemed to be important were weighed and combined to create a Confidence Index (CI): ((r2)4 x 6) x ((Q2)4 x 6) x (ln(Nc/10)) x (Tr) x (Sconf)0.5 (ln(N2d + 2)) x (ln(N2m + 2)) x ((r2)4 x 6) - ((Q2)4 x 6) + 1) x (Rconf)

Algorithms↗