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At least 523 records · Page 29Linked to original sources

3-D particle tracking in a two-photon microscope: application to the study of molecular dynamics in cells.

We developed a method for tracking particles in three dimensions designed for a two-photon microscope, which holds great promise to study cellular processes because of low photodamage, efficient background rejection, and improved depth discrimination. During a standard cycle of the tracking routine (32 ms), the laser beam traces four circular orbits surrounding the particle in two z planes above and below the particle. The radius of the orbits is half of the x,y-width of the point spread function, and the distance between the z planes is the z-width of the point spread function. The z-position is adjusted by moving the objective with a piezoelectric-nanopositioner. The particle position is calculated on the fly from the intensity profile obtained during the cycle, and these coordinates are used to set the scanning center for the next cycle. Applying this method, we were able to follow the motion of 500-nm diameter fluorescent polystyrene microspheres moved by a nanometric stage in either steps of 20-100 nm or sine waves of 0.1-10 microm amplitude with 20 nm precision. We also measured the diffusion coefficient of fluorospheres in glycerol solutions and recovered the values expected according to the Stokes-Einstein relationship for viscosities higher than 3.7 cP. The feasibility of this method for live cell measurements is demonstrated studying the phagocytosis of protein-coated fluorospheres by fibroblasts.

Animals↗

A method for quantifying differential expansion within hydrating hydrophilic matrixes by tracking embedded fluorescent microspheres.

A method is described for quantifying the pattern of deformation within a matrix and is demonstrated by analyzing the expansion of polymer hydrophilic matrix tablets. The fundamental features of the method are the incorporation of nondiffusing markers into the matrix and the subsequent tracking of these markers during deformation. Since the markers are too large to diffuse, their individual movement reflects the translocation of the surrounding matrix, and the separation between pairs of markers reveals any perturbation in the intervening area. By tracking many markers, the pattern of deformation within a matrix can be ascertained. The method was demonstrated on hydrating hydrophilic matrix tablets, using fluorescent microspheres as nondiffusing markers which were observed with a confocal laser scanning microscope. Analysis of the tracks showed a wave of expansion moving from the exterior toward the core, with the greatest and earliest expansion found in the outer regions. The results also showed that even as deeper layers started to expand the outer layers continued to swell.

Chemistry, Pharmaceutical↗

Novel antigen system for tracking epithelial cell migration in serum-free culture.

We report here a unique system for tracking normal human urothelial cell migration in serum-free culture medium (HMRI-1). The key observation was that urothelial cells deposited red blood cell surface antigen on the culture dish in a remarkable pattern. Scrutiny of this pattern showed that each migrating cell left behind antigen imprints which formed parallel tracks the width of the cell. Hence the previous migratory history of the cells was instantly mapped by simply visualizing the antigen tracks deposited by the cells on the dish. Apart from providing a simple method for tracking urothelial cells, this observation has wider implications for mechanistic studies of epithelial cell movement in general. It also highlights the complicating effects associated with the addition of serum as a traditional culture supplement, since the inclusion of serum in the HMRI-1 medium abolished the above effect by inhibiting cell migration.

ABO Blood-Group System↗

Flow cytometric analysis of platelets.

BACKGROUND AND OBJECTIVES: Despite recent progress in our understanding of platelet function in vitro, there remains a remarkable paucity of methods to study platelet function in vivo. MATERIALS AND METHODS: We have developed novel three color whole blood flow cytometric methods for tracking of infused platelets and measurement of their function in vivo. RESULTS: These methods were used to demonstrate that circulating P-selectin-positive (degranulated) platelets rapidly lose surface P-selectin to the plasma pool, but continue to circulate and function. CONCLUSIONS: 1) These studies strongly suggest that the measurement of platelet surface P-selectin in platelet concentrates stored in the blood bank should not be used as a predictor of platelet survival or function in vivo. 2) The described methods provide the means to answer many previously difficult to address questions about in vivo platelet function in transfusion medicine.

Animals↗

Improving performance of distribution tracking through background mismatch.

This paper proposes a new density matching method based on background mismatching for tracking of nonrigid moving objects. The new tracking method extends the idea behind the original density-matching tracker, which tracks an object by finding a contour in which the photometric density sampled from the enclosed region most closely matches a model density. This method can be quite sensitive to the initial curve placements and model density. The new method eliminates these sensitivities by adding a second term to the optimization: The mismatch between the model density and the density sampled from the background. By maximizing this term, the tracking algorithm becomes significantly more robust in practice. Furthermore, we show the enhanced ability of the algorithm to deal with target objects which possess smooth or diffuse boundaries. The tracker is in the form of a partial differential equation, and is implemented using the level-set framework. Experiments on synthesized images and real video sequences show our proposed methods are effective and robust; the results are compared with several existing methods.

Algorithms↗

Real-time tracking using trust-region methods.

Optimization methods based on iterative schemes can be divided into two classes: line-search methods and trust-region methods. While line-search techniques are commonly found in various vision applications, not much attention is paid to trust-region ones. Motivated by the fact that line-search methods can be considered as special cases of trust-region methods, we propose to establish a trust-region framework for real-time tracking. Our approach is characterized by three key contributions. First, since a trust-region tracking system is more effective, it often yields better performances than the outcomes of other trackers that rely on iterative optimization to perform tracking, e.g., a line-search-based mean-shift tracker. Second, we have formulated a representation model that uses two coupled weighting schemes derived from the covariance ellipse to integrate an object's color probability distribution and edge density information. As a result, the system can address rotation and nonuniform scaling in a continuous space, rather than working on some presumably possible discrete values of rotation angle and scale. Third, the framework is very flexible in that a variety of distance functions can be adapted easily. Experimental results and comparative studies are provided to demonstrate the efficiency of the proposed method.

Algorithms↗

Kinetic analysis of the mitotic cycle of living vertebrate cells by atomic force microscopy.

The atomic force microscope (AFM) is becoming an important tool for qualitative and quantitative analyses of biological material. However, the difficulties involved in maintaining long-term, steady-state physiologic conditions and the problems associated with analyzing force curves generated from highly viscoelastic biological structures impede the use of the AFM for studies of kinetic processes in living vertebrate cells. In this report, we describe a simple method to track reproducibly kinetic changes in the localized stiffness of vertebrate cells. We tested our method on a study of vertebrate cells in mitosis and found a marked but transient decrease in stiffness occurs in the mitotic spindle region during anaphase. We propose that physical-chemical changes in the mitotic apparatus, most probably, changes in the state of polymerization of interzonal spindle fibers which also have been reported to undergo a marked reduction in birefringence during anaphase, are responsible for the observed decrease in stiffness. Our methodology affords a new approach to studying mitotic events and should be applicable to studies of a variety of viscoelastic properties of living cells.

Anaphase↗

Diagnostic errors.

One of the most complex and challenging tasks facing physicians is the need to make a diagnosis. Recent research has focused attention on medical errors alleged to have resulted in increased patient morbidity and mortality. A number of authorities have suggested methods to track and prevent errors. Most have dealt with systems' changes and fail-safe methods to prevent medication errors and commission of errors during treatment. Few have addressed methods to find and prevent diagnostic errors. Unless diagnostic error is either avoided or corrected early, fail-safe methods to prevent medication and treatment errors will ultimately fail to improve patient outcome. American medical literature, particularly postmortem studies, have documented diagnostic error since at least 1912. European literature shows the problem is worldwide. The limits of human memory and errors in both observation and processing of information during problem solving contribute to the commission of errors. The purpose of this article is to examine the thinking patterns and cognitive errors that can result in diagnostic error, and suggest instructional strategies that can be used to alert residents and attending physicians to these potential problems so they can be avoided.

Diagnostic Errors↗

Fluorescent derivatization of a protease antigen to track antigen uptake and processing in human cell lines.

BACKGROUND: We have devised a simple and efficient fluorescence-based method to track antigen uptake and processing in human B lymphoblastoid cells (B-LCL). Fluorescein labelled subtilisin was used to optimize antigen uptake conditions and identify processed peptides from human cell lines. RESULTS: Fluorescein labelled subtilisin conjugates had 0.06 to 2 moles of fluorescein per subtilisin molecule. High performance liquid chromatography and mass spectrometry (NanoESI-LC/MS/MS) analysis identified fluorescein conjugated to K141, K256, and the N terminus. Conjugates retained antigenic specificity to subtilisin specific antibodies and could be processed by whole cell extracts into low molecular weight fragments at pH 5.2. Maximal antigen uptake and processing occurred when PMSF (phenylmethylsulfonyl fluoride) inhibited subtilisin conjugate was incubated with cells at 100-200 microg/ml for 16 to 24 hr. Once optimal uptake conditions were established, processed subtilisin peptides were isolated and identified from human cell lines. CONCLUSION: Our studies show that FITC-conjugation provides an efficient tool to track the uptake and processing of this protease antigen and to facilitate identification of processed antigenic peptides from human cell lines.

Antigens↗

Projection flow imaging by bolus tracking using stimulated echoes.

Previous investigators have employed the concept of bolus tracking using either spin echoes or gradient echoes. In this paper we introduce two methods of bolus tracking using planar- and volume-selective stimulated echoes. The planar method employs a selective 90 degrees rf pulse which tags all spins in a particular plane. At a time tau 1 later, a nonselective 90 degrees rf pulse is employed, followed after a time tau 2, by another nonselective rf pulse. Only spins which experience all three rf pulses form a stimulated echo at time tau 1 after the third rf pulse. A balanced pair of flow-compensated dephasing (crusher) gradients further ensures that the stimulated echo is due only to the effect of all three rf pulses while minimizing flow dephasing. The first part of this gradient pair is applied after the initial rf pulse in the first tau 1 period to dephase the tagged spins. The second part of this gradient pair is applied after the third rf pulse to rephase the spins. Since the plane of the excited slice is orthogonal to the readout direction, flowing spins are imaged in an angiographic manner as they move away from the excited slice. A modification to this basic sequence excites only a small volume. In this manner, the suppression of stationary spins is effected by volume-selective excitation. In both the planar- and the volume-selective techniques, the excited spins undergo T1 and T2 relaxation during the tau 1 period but only T1 relaxation in the tau 2 period. In blood, where T1 is much greater than T2, keeping tau 1 as short as possible minimizes signal loss due to T2 dephasing. These methods demonstrate increased sensitivity compared to similar bolus tracking methods using either spin echoes or gradient echoes.

Blood Flow Velocity↗

Potential use of a host associated molecular marker in Enterococcus faecium as an index of human fecal pollution.

Several genotypic and phenotypic microbial source tracking (MST) methods have been proposed and utilized to differentiate groups of microorganisms, usually indicator organisms, for the purpose of tracking sources of fecal pollution. Targeting of host-specific microorganisms is one of the approaches currently being tested. These methods are useful as they circumvent the need to isolate individual microorganisms and do not require the establishment of reference databases. Several studies have demonstrated that the presence and distribution of Enterococcus spp. in feces seems to be influenced by the host species. Here, we present a method for detection of genetic sequences in culturable enterococci capable of identifying human sources of fecal pollution in the environment. The human fecal pollution marker designed in this study targets a putative virulence factor, the enterococcal surface protein (esp), in Enterococcus faecium. This gene was detected in 97% of sewage and septic samples but was not detected in any livestock waste lagoons or in bird or animal fecal samples. Epidemiological studies in recreational and groundwaters have shown enterococci to be useful indicators of public health risk for gastroenteritis. By identifying the presence of human fecal pollution, and therefore the possible presence of human enteric pathogens, this marker allows for further resolution of the source of this risk.

DNA, Bacterial↗

Computer-monitored radionuclide tracking of three-dimensional mandibular movements. Part II: experimental setup and preliminary results--Posselt diagram.

This article described a new method to track mandibular movements using a computer-assisted radionuclide kinematics technique. The usefulness of various image-enhancement techniques is discussed, and the reproduction of physiologic displacements is shown. Vertical, lateral, and protrusive envelopes of motion of a point on a tooth of a complete denture mounted on a semiadjustable articulator were measured. A demonstrative example of the validity of this approach is reproducing the motion of the mental point, which clearly evidences the Posselt diagram.

Computers↗

Weight loss composition: the effects of exercise following obesity surgery as measured by bioelectrical impedance analysis.

BACKGROUND: Sudden weight loss following bariatric operations for morbid obesity, such as the duodenal switch (DS), can result in a concurrent decrease in lean body mass. Several methods for tracking body composition, such as bioelectrical impedance analysis (BIA), are available to monitor these changes. One method to offset the negative effects of sudden weight loss on body mass composition may be exercise. METHODS: 100 patients who had undergone the DS operation for morbid obesity were classified as exercisers and non-exercisers based on self-reporting. Their body mass compositions were measured using BIA preoperatively and at 0.75, 1.5, 3, 6, 9, 12, and 18 months postoperatively. RESULTS: At no study interval did postoperative percent changes in weight loss differ between the exercise and non-exercise groups. At 18 months postoperatively, the exercise group showed a 28% higher loss of fat mass and an 8% higher gain in lean body mass than the non-exercise group. CONCLUSION: Exercise positively influences body mass composition following the DS. BIA can be successfully employed to monitor changes, diagnose deficiencies, and formulate treatment recommendations.

Adult↗

Can we track the impact of Australian mental health research?

OBJECTIVE: Arguments are being made to increase research and development funding for mental health research in Australia. Consequently, the methods used to measure the results of increased investment require review. This study aimed to describe the status of Australian mental health research and to propose potential methods for tracking changes in research output. Specifically, we describe the research output of nations, Australian states, Australian and New Zealand institutions and Australian and New Zealand researchers using citation rates. METHOD: Information on research output was sourced from two international databases (Institute for scientific information [ISI] Essential Science Indicators and ISI Web of Science) and the ISI list of Highly Cited Researchers. RESULTS: In an international setting, Australia does not perform as well as other comparable countries such as New Zealand or Canada in terms of research output. Within Australia, the scientific performance of institutions apparently relates to the strength of some individual researchers or consolidated research groups. Highly cited papers are evident in the fields of syndrome definition, epidemiology and epidemiological methods, cognitive science and prognostic or longitudinal studies. CONCLUSIONS: Australian researchers need to consider the success of New Zealand and Canadian researchers, particularly given the relatively low investment in health and medical research in New Zealand. Although citation analyses are fraught with difficulties, they can be effectively complemented by other measures of responsiveness to clinical or population needs and community expectations and should be conducted regularly and independently to monitor the status of Australian mental health research.

Australia↗

Method for combining information from white matter fiber tracking and gray matter parcellation.

We introduce a method for combining fiber tracking from diffusion-tensor (DT) imaging with cortical gray matter parcellation from structural high-spatial-resolution 3D spoiled gradient-recalled acquisition in the steady state images. We applied this method to a tumor case to determine the impact of the tumor on white matter architecture. We conclude that this new method for combining structural and DT imaging data is useful for understanding cortical connectivity and the localization of fiber tracts and their relationship with cortical anatomy and brain abnormalities.

Adult↗

Three-dimensional localization of sperm whales using a single hydrophone.

A three-dimensional localization method for tracking sperm whales with as few as one sensor is demonstrated. Based on ray-trace acoustic propagation modeling, the technique exploits multipath arrival information from recorded sperm whale clicks and can account for waveguide propagation physics like interaction with range-dependent bathymetry and ray refraction. It also does not require ray identification (i.e., direct, surface reflected) while utilizing individual ray arrival information, simplifying automation efforts. The algorithm compares the arrival pattern from a sperm whale click to range-, depth-, and azimuth-dependent modeled arrival patterns in order to estimate whale location. With sufficient knowledge of azimuthally dependent bathymetry, a three-dimensional track of whale motion can be obtained using data from a single hydrophone. Tracking is demonstrated using data from acoustic recorders attached to fishing anchor lines off southeast Alaska as part of efforts to study sperm whale depredation of fishing operations. Several tracks of whale activity using real data from one or two hydrophones have been created, and three are provided to demonstrate the method, including one simultaneous visual and acoustic localization of a sperm whale actively clicking while surfaced. The tracks also suggest that whales' foraging is shallower in the presence of a longline haul than without.

Acoustics↗

Excitation-emission matrix fluorescence spectroscopy in conjunction with multiway analysis for PAH detection in complex matrices.

A field portable, single exposure excitation-emission matrix (EEM) fluorometer has been constructed and used in conjunction with parallel factor analysis (PARAFAC) to determine the sub part per billion (ppb) concentrations of several aqueous polycyclic aromatic hydrocarbons (PAHs), such as benzo(k)fluoranthene and benzo(a)pyrene, in various matrices including aqueous motor oil extract and asphalt leachate. Multiway methods like PARAFAC are essential to resolve the analyte signature from the ubiquitous background in environmental samples. With multiway data and PARAFAC analysis it is shown that reliable concentration determinations can be achieved with minimal standards in spite of the large convoluting fluorescence background signal. Thus, rapid fieldable EEM analyses may prove to be a good screening method for tracking pollutants and prioritizing sampling and analysis by more complete but time consuming and labor intensive EPA methods.

Journal Article↗

RNA dynamics in live Escherichia coli cells.

We describe a method for tracking RNA molecules in Escherichia coli that is sensitive to single copies of mRNA, and, using the method, we find that individual molecules can be followed for many hours in living cells. We observe distinct characteristic dynamics of RNA molecules, all consistent with the known life history of RNA in prokaryotes: localized motion consistent with the Brownian motion of an RNA polymer tethered to its template DNA, free diffusion, and a few examples of polymer chain dynamics that appear to be a combination of chain fluctuation and chain elongation attributable to RNA transcription. We also quantify some of the dynamics, such as width of the displacement distribution, diffusion coefficient, chain elongation rate, and distribution of molecule numbers, and compare them with known biophysical parameters of the E. coli system.

Diffusion↗