Evaluation of the usability of a brief computerized cognitive screening test in older people for epidemiological studies.

BACKGROUND: Computerized cognitive testing has the potential to be an effective way to assess and monitor cognition in large neuroepidemiological studies. CogState is a game-like computerized test with demonstrated validity and reliability that has shown sensitivity to decline in older individuals over time. This study aimed to evaluate the serial usability of the test specifically within an older community cohort. METHODS: The test battery was administered to healthy volunteers aged 50 years and above at 3-month intervals over 12 months in a community setting. Test usability was examined in terms of acceptability, efficiency and stability. RESULTS: Of 301 subjects (age: 61.9 +/- 7.2 years), 87% completed the study. In addition, 85% completed the first test within the allowed time and passed integrity criteria with their performance improving and stabilizing at subsequent visits. The computerized battery required 15 min for administration on average, allowing 263 patients to be assessed on 5 occasions by 2 assessors. All tasks showed stability and a high test-retest reliability with serial administration. CONCLUSIONS: This computerized test was shown to have good acceptability, efficiency and stability for the repeated assessment of cognitive function in older people. Together with its demonstrated sensitivity to cognitive impairment and cognitive change, these data suggest that it would be a useful tool for application in neuroepidemiological studies.

Metabolites of a tobacco-specific lung carcinogen in the urine of elementary school-aged children.

Limited data are available in the literature on carcinogen uptake by children exposed to environmental tobacco smoke (ETS). In this study, we quantified metabolites of the tobacco-specific lung carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in the urine of elementary school-aged children participating in the School Health Initiative: Environment, Learning, Disease study, a school-based investigation of the environmental health of children. The metabolites of NNK are 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and its glucuronide (NNAL-Gluc). We also measured cotinine and its glucuronide (total cotinine). Urine samples were collected from 204 children. Seventy (34.3%) of these had total cotinine > or =5 ng/ml. NNAL or NNAL-Gluc was detected in 52 of 54 samples with total cotinine > or =5 ng/ml and in 10 of 20 samples with total cotinine < 5 ng/ml. Levels of NNAL plus NNAL-Gluc and total cotinine were significantly higher when exposure to ETS was reported than when no exposure was reported. However, even when no exposure to ETS was reported, levels of NNAL, NNAL-Gluc, and NNAL plus NNAL-Gluc were higher than in children with documented low exposure to ETS, as determined by cotinine levels < 5 ng/ml. Levels of NNAL, NNAL-Gluc, and cotinine were not significantly different in samples collected twice from the same children at 3-month intervals. Levels of NNAL plus NNAL-Gluc in this study were comparable with those observed in our previous field studies of adults exposed to ETS. There was a 93-fold range of NNAL plus NNAL-Gluc values in the exposed children. The results of this study demonstrate widespread and considerable uptake of the tobacco-specific lung carcinogen NNK in this group of elementary school-aged children, raising important questions about potential health risks. Our data indicate that objective biomarkers of carcinogen uptake are important in studies of childhood exposure to ETS and cancer later in life.

A school-based strategy to assess children's environmental exposures and related health effects in economically disadvantaged urban neighborhoods.

The School Health Initiative: Environment, Learning, Disease (SHIELD) study is a novel school-based investigation of children's environmental health in economically disadvantaged urban neighborhoods of Minneapolis. This article describes the study design and summarizes lessons learned about recruiting and monitoring this historically understudied population. The SHIELD study focused on measuring children's exposures to multiple environmental stressors [volatile organic chemicals (VOCs), environmental tobacco smoke, allergens, bioaerosols, metals, pesticides, polychlorinated biphenyls (PCB), phthalates] and exploring related effects on respiratory health (e.g., lung function) and learning outcomes (e.g., standardized test scores, academic achievement). It involved intensive exposure monitoring, including environmental measurements inside and outside the children's schools and inside their homes, personal measurements with passive dosimeters worn by the children, and biological marker measurements in blood and urine. The SHIELD participants comprised a stratified random sample of 153 "index" children and 51 of their siblings enrolled in grades 2-5 at two adjacent elementary schools. The Minneapolis Public Schools (MPS) assisted with identifying, contacting, recruiting, and monitoring this population, which traditionally is difficult to study because families/children are highly mobile, speak a diversity of languages, frequently do not have a telephone, endure economic hardships, often do not trust researchers, and have a spectrum of unconventional lifestyles and living arrangements. Using a school-based approach, the overall SHIELD enrollment (response) rate was 56.7%, with a wide disparity between English-speaking (41.7%) and non-English-speaking (71.0%) families/children. Most children remained involved in the study through both monitoring sessions and exhibited an acceptable degree of compliance with study protocols, including providing blood and urine samples. Results indicate that it is both practical and affordable to conduct probability-based exposure studies in this population, but that it is also important to improve our understanding of factors (e.g., cultural, economic, psychological, social) affecting the willingness of families/children to participate in such studies, with special emphasis on developing cost-effective recruitment methods.

Selective synchronization of Tetrahymena pyriformis cell populations and cell growth kinetics during the cell cycle.

A selection synchronization technique based on ingestion of tantalum particles has been applied to obtain synchronized cultures of the filter feeding ciliate Tetrahymena pyriformis. Cell concentrations and cell volume distributions of synchronized cell populations have been monitored for more than four average generation times. A simple curve-fitting method has been used to decompose the cell volume distributions of a synchronous population into two populations representing cells before and after cell division. In this way, the time course of the growth of an initially synchronous culture is decomposed into the growth of successive generations. The data indicate that at any given time only two generations of cells are present in significant numbers. The measured cell volume distributions show that T. pyriformis has a complicated growth pattern during the cell cycle. Newborn T. pyriformis cells do not grow significantly at the beginning of the cell cycle. After the nongrowing stage, cells start growing in a possibly exponential rate before cells enter into a second nongrowing stage. The second nongrowing stage lasts until cell division. The presented data demonstrate that growing cell populations can be viewed as the composite of cells belonging to different generations. This concept has important implications for solving corpuscular models of cell growth.

The effect of non-insulin-dependent diabetes mellitus and obesity on glucose transport and phosphorylation in skeletal muscle.

Defects of glucose transport and phosphorylation may underlie insulin resistance in obesity and non-insulin-dependent diabetes mellitus (NIDDM). To test this hypothesis, dynamic imaging of 18F-2-deoxy-glucose uptake into midthigh muscle was performed using positron emission tomography during basal and insulin-stimulated conditions (40 mU/m2 per min), in eight lean nondiabetic, eight obese nondiabetic, and eight obese subjects with NIDDM. In additional studies, vastus lateralis muscle was obtained by percutaneous biopsy during basal and insulin-stimulated conditions for assay of hexokinase and citrate synthase, and for immunohistochemical labeling of Glut 4. Quantitative confocal laser scanning microscopy was used to ascertain Glut 4 at the sarcolemma as an index of insulin-regulated translocation. In lean individuals, insulin stimulated a 10-fold increase of 2-deoxy-2[18F]fluoro-D-glucose (FDG) clearance into muscle and significant increases in the rate constants for inward transport and phosphorylation of FDG. In obese individuals, the rate constant for inward transport of glucose was not increased by insulin infusion and did not differ from values in NIDDM. Insulin stimulation of the rate constant for glucose phosphorylation was similar in obese and lean subjects but reduced in NIDDM. Insulin increased by nearly twofold the number and area of sites labeling for Glut 4 at the sarcolemma in lean volunteers, but in obese and NIDDM subjects translocation of Glut 4 was attenuated. Activities of skeletal muscle HK I and II were similar in lean, obese and NIDDM subjects. These in vivo and ex vivo assessments indicate that impaired glucose transport plays a key role in insulin resistance of NIDDM and obesity and that an additional impairment of glucose phosphorylation is evident in the insulin resistance of NIDDM.

Multistaged corpuscular models of microbial growth: Monte Carlo simulations.

A new framework is developed by extending the existing population balance framework for modeling the growth of microbial populations. The new class of multistaged corpuscular models allows further structuring of the microbial life cycle into separate phases or stages and thus facilitates the incorporation of cell cycle phenomena to population models. These multistaged models consist of systems of population balance equations coupled by appropriate boundary conditions. The specific form of the equations depend on the assumed forms for the transition rate functions, the growth rate functions, and the partitioning function, which determines how the biological material is distributed at division. A growth model for ciliated protozoa is formulated to demonstrate the proposed framework. To obtain a solution to the system of the partial integro differential equations that results from such formulation, we adopted a Monte Carlo simulation technique which is very stable, versatile, and insensitive to the complexity of the model. The theory and implementation of the Monte Carlo simulation algorithm is analyzed and results from the simulation of the ciliate growth model are presented. The proposed approach seems to be promising for integrating single-cell mechanisms into population models.

Feeding heterogeneity in ciliate populations: Effects of culture age and nutritional state.

We have used a novel approach in conjunction with flow cytometry to quantify the biological heterogeneity of populations of the ciliate Tetrahymena pyriformis. It was found that the rate of particle uptake of exponentially growing cells is not uniform among cells and partially correlated with cell size. The physiological state and growth history of the culture was found to affect to a large degree the population's feeding heterogeneity. Stationary phase populations exhibited more uniform feeding behavior, as cell aging affects all cells and effectively reduces their ability to feed. Cells that were removed from the growth medium and resuspended in nonnutritive medium exhibited a more heterogeneous feeding behavior. The starved cells were stimulated to feed at considerably higher rates, and the stimulatory effect was more pronounced for larger cells. It is therefore demonstrated that population heterogeneity has to be evaluated in conjunction with the populations growth state as it is determined by the history of the population's growth and nutritional state. (c) 1994 John Wiley & Sons, Inc.

Determination of cellular rate distributions in microbial cell populations: feeding rates of ciliated protozoa.

A novel procedure is proposed for determining distributions of rate properties and correlations of rate with state properties of microbial cell populations. The procedure is novel in that it uses transient data, and thus, it does not require that the population be in balanced growth, although it requires that the population structure does not change during the short transient experiment. The procedure is applied to populations of the ciliated protozoan Tetrahymena to determine ingestion rate variability. The number of ingested microspheres per cell and the single-cell protein content-an indicator of cell size-were directly determined with dual-color flow cytometry. The proposed technique revealed the correlation pattern of the particle ingestion rate with cell size. In particular, ingestion rate was found to be positively correlated with cell size for the smaller feeding cells and to be uncorrelated with size for the larger cells. Using the fact that particle uptake from dilute particle suspensions is a Poisson random process, we determined that the coefficient of variation of the distribution of ingestion rates within the feeding population is about 50%. It was concluded that the dynamics of particle ingestion can be accurately described only if it is realized that particle ingestion rates are distributed.

A statistical analysis of flow cytometric determinations of phagocytosis rates.

Uptake of particles by phagocytosing cells is a process that exhibits variability of its rate. This variability is inherent in the mechanism of particle uptake and in the mechanisms that determine the distribution of physiological states within a population of phagocytosing cells. When numbers of particles ingested by cells are determined flow cytometrically an additional measurement variability is superimposed on and interacts with the aforementioned biological variability. In one method of determining population phagocytosis parameters, which involves fitting theoretical equations to experimental time course data on the fractions of cells which have ingested 0, 1, 2, ... particles, the effects of measurement variability are circumvented, although this usually has the cost of not using all the sample data obtained. However, in a second, simpler, method which is based on determining the time course of the number of particles ingested by an average cell, measurement variability is not circumvented and its effects must be considered. An analysis of the combined effects of biological and measurement variability on the results obtained with the simpler method is presented in this paper. Experimental results for phagocytosis of latex microspheres of uniform size and fluorochrome content by populations of the ciliate Tetrahymena pyriformis show that, for this system, measurement variability is entirely negligible in comparison with biological variability. This conclusion might not apply to other systems, however, and situations which might make measurement variability of some significance are mentioned in the paper. The equations given can be used for the analysis of such situations.

Segregated, structured, distributed models and their role in microbial ecology: A case study based on work done on the filter-feeding ciliateTetrahymena pyriformis.

Microbial populations are composed of individual organisms each of which, if environmental circumstances are favorable, is undergoing change of its internal state through the operation of the set of processes that we call the cell cycle. The rate of progression through the cycle is subject to internal controls as well as external influences, and exhibits random as well as deterministic features. Microorganisms of the same species in different stages of the cell cycle have different internal states, and thus, the operation of the cell cycle is by itself sufficient to produce a distribution of states among the individual organisms of a population. In turn, the distribution of states produces distributions of the rates at which the cells of a population carry on their activities. Mathematical models of microbial growth that take the operation of the cell cycle and its consequences into account are more complicated than the kinds of models that are often used in microbial ecology. This paper gives some account of the nature, formulation, and uses of complex growth models. The account is illustrated by work done by the author and his collaborators H. M. Tsuchiya and more recently F. Srienc, as well as by others, on the filter-feeding ciliateTetrahymena pyriformis.

A discrete, stochastic model for microbial filter feeding: a model for feeding of ciliated protists on spatially uniform, nondepletable suspensions.

Suspension-feeding ciliates, either bacteriovorous or planktonic, are adapted to feed on particulate food matter of size much smaller than their own size. These microorganisms collect their prey by generating water currents that draw prey toward their capture surfaces. Under such conditions food particles are treated in bulk, and captures of individual food particles from a suspension by individual single-celled organisms are discrete events that occur at random intervals of time. Each such event is followed by a sequence of additional events that also occur at random intervals of time. This sequence culminates in the incorporation of the digestible portion of the food particle into the cell's cytoplasm and the expulsion of the indigestible portion from the cell. In theory, the rate of the overall ingestion-digestion process can be limited by the passage of particles through any stage of this sequence of events. In this paper, we assume that only the initial events in the sequence, those that occur in the oral region of the cell, limit the rate of the ingestion-digestion process, and we develop a discrete, stochastic model of filter feeding based on that assumption. We use the model to show how advanced instrumentation, such as flow cytometry, can be used to measure parameters of the model and also to answer a number of important questions about the mechanism of filter feeding. We show also how the model can be applied to nonhomogeneous cell populations for which parameters of the model are distributed.

Flow cytometric measurement of rates of particle uptake from dilute suspensions by a ciliated protozoan.

Flow cytometry is used to measure rates of ingestion of particles from dilute monodisperse suspensions by the ciliate Tetrahymena pyriformis. The particles used are polystyrene microspheres containing a fluorescent dye. Measurements were made directly, that is, by determining the fluorescence intensities from microspheres ingested by cells in samples collected from the experimental feeding apparatus. The fact that fluorescence intensities from individual cells can be grouped into discrete classes based on the numbers of fluorescent particles associated with the cells makes it possible to calibrate the flow cytometer and convert fluorescence measurements into numbers of particles ingested by average cells. At low particle concentration or high ciliate concentration, ingestion data must be corrected for depletion of particles during the assay, and a method for doing this is described. Experiments at various ciliate concentrations show that ingestion rates are not affected by this concentration. The methods developed should allow measurements of rates of ingestion of particles from concentrated and polydisperse suspensions. For such measurements, nonfluorescent particles together with a fraction of fluorescent tracer particles would be used.

Growth of microbial populations in nonminimal media: some considerations for modeling.

Ramkrishna and his co-workers have developed so-called cybernetic models which purport to describe, among other things, how microorganisms make choices when presented with two or more functionally equivalent, or substitutable, nutrients that are sources of carbon and available energy. In general, however, organisms are presented with choices not just between nutrients that are substitutable for one another, but also between sets of nutrients some of which are by no means substitutable for one another. It is postulated herein that the main ideas of cybernetic modeling apply to these more general choices as they seem to apply to the choices considered by Ramkrishna and his co-workers. Some consequences of the postulate are worked out for steady-state growth situations where two, or in one case three, nutrients limit or potentially limit growth rate. If predicted phenomena are observed experimentally so as to verify the postulate, a significantly improved basis for understanding growth of microorganisms in practical fermentation media as well as in natural situations will be provided by this application of cybernetic modeling ideas.

Clinical and analytical evaluation of two immunoassays for direct measurement of creatine kinase MB with monoclonal anti-CK-MB antibodies.

We examined the clinical and analytical performance of two immunoassays (Becton Dickinson CK-MB; Ciba-Corning Magic Lite CK-MB) in which monoclonal anti-CK-MB antibodies are used for directly measuring creatine kinase (EC 2.7.3.2) isoenzyme MB (CK-MB) in serum, and also one electrophoretic method (Ciba-Corning). Within- and between-assay precision for both immunoassays was good at the upper reference limits (less than 10% CV). Analytical recoveries ranged from 102 to 114%. Both immunoassays were free from interference by CK-BB, mitochondrial-CK, macro-CK, adenylate kinase, and CK-MM. Retrospectively, we evaluated four categories of patients, using both immunoassays and electrophoresis: normal controls, acute myocardial infarction (AMI) patients, severe skeletal muscle trauma patients, and acutely ill patients known not to have AMI. In general, there were excellent correlations among all three methods. CK-MB activity (U/L) measured by the Becton Dickinson immunoassay was approximately 50% of the mass concentration (microgram/L) of the Magic Lite immunoassay and 50% of the activity concentration (U/L) determined by electrophoresis. Both immunoassays were easy to perform and sensitive to the low CK-MB concentrations often found with low total-CK activities.

Limitation of growth rate by two complementary nutrients: some elementary but neglected considerations.

Experimental data in the literature show that the yield of biomass from a particular nutrient when that nutrient limits growth rate is often significantly different than the yield from the nutrient when some complementary nutrient limits growth rate. This article explores some possible consequences for bioreactor dynamics of dependence of yield coefficients on the identity of the nutrient that limits growth rate.

Peristance of bacteria in the presence of viable, nonencysting, bacterivorous ciliates.

Laboratory studies of the interactions between a bacterial population and a population of bacterivorous ciliates consistently show that the bacteria are able to persist in the presence of viable ciliates. Reproduction of the bacteria, presumably at the expense of substrates produced by death and lysis of the ciliates and/or by their metabolic activity, has been suggested to be a factor involved in the observed bacterial persistence. Rates and extents of growth ofEscherichia coli in broths of mixed cultures of this bacterium and the ciliateTetrahymena pyriformis were determined in order to provide some data necessary to assess the importance of the suggested factor. In addition, an attempt was made to suppress bacterial growth on produced substrates so that feeding of the ciliates could be studied free of this complication. However, the procedure tested-addition of the antibiotic chloramphenicol (CM) at a concentration of 150µg/ml-led to other complications that made it impossible to obtain the desired information about feeding.

Coexistence of bacteria and feeding ciliates: Growth of bacteria on autochthonous substrates as a stabilizing factor for coexistence.

There is good evidence that the specific feeding and growth rates of a population of suspension-feeding ciliates on a population of bacteria is described by the classic Monod model or something very like it. However, use of this model combined with the assumption that feeding of the ciliates is the only interaction between the populations leads to predictions grossly at variance with experimental observations. A second interaction between the populations, growth of the bacteria on products of lysis, and perhaps metabolism of the ciliates is known to occur and might be an important factor in determining the dynamics of ciliate-bacterial systems. Whether or not this is the case is examined in the present article. It is concluded that accounting for the second interaction does not remove the discrepancies between model predictions and experimental observations. It is concluded also that the second interaction is likely not important except in batchtype situations where the supply of available energy for supporting biological activity is not replenished.