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Platelet estimation in whole blood.

An application of particle size analysis is described for the estimation of platelet counts in whole blood based on a variable size threshold and baseline. The simplicity of the sample preparation for this technique may enable its automation as a semiquantitative screening method. Platelets, defined as particles to the platelet side and above the minimum point between the platelet and red cell distributions were counted on specially designed electronics. The system divides the normal size distribution from the smallest platelet to the smallest red cell into four equal windows and performs three integrations to calculate the best result. Correlation of results obtained using this technique with results form the sedimentation method produced a coefficient of 0.93. In the trial period of a week in which all the samples (132) normally undergoing platelet count were also counted on the system descirbed, only six false negative results were found, four at the lower end of normal (150000) and two at the upper end (400000). The advantages of an MCA computer-based system are also discussed.

Blood Cell Count↗

A comparison of optimum detector spatial resolution in nuclear imaging based on statistical theory and on observer performance.

An expression for the expected image of a spherical tumour in a uniform background was derived in terms of background thickness and concentration of radioactivity, the tumour size, depth and uptake ratio, the gamma-ray energy and the detector response function. Three models of human observer performance for tumour detection were developed from different signal-to-noise ratio measures based on the statistical theory of detection. The optimum detector spatial resolution predicted by each model was then compared to that obtained from an observer performance study in which the subjects viewed computer-simulated scintigrams. The predictions from two of these models seem to be consistent with the results of the observer performance study. Model II involves a comparison of the counts integrated over the tumour region with the counts integrated over a background region of the same area. Model III compares the count density estimates of signal-plus-background and background obtained from application of non-uniform weighting functions to the image data.

Gamma Rays↗

Measurement of low neutron fluences with polycarbonate foils electrochemically etched with methyl alcohol-KOH solution.

Electrochemical etching of polycarbonate foils was performed using a KOH solution with a high concentration of methyl alcohol under different conditions of field strength, frequency, temperature and etching time. these studies showed that the highest neutron sensitivity relative to the inherent background in the foil was obtained under the following etching conditions: 52 kV/cm, 1 kHz, 30 degrees C, 30 min in a solution of 45 gm KOH + 80 cc CH3OH + 20 cc H2O. Under these conditions, 100 mrem of neutrons from a Ra-Be source gave 70 pits per cm2 while background was 7 +/- 3 pits per cm2 (10 +/- 5 mrem). The pit diameters were about 90 micrometers. This sensitivity (giving lowest measurable dose of 15 mrem) is quite sufficient for personnel neutron dosimetry applications and the size of the pits is large enough for easy counting using a microscope at a magnification of 40 X.

Electrochemistry↗

Evaluation of an alpha probe detector for in vitro cellular dosimetry.

This paper describes the design and testing of an alpha probe detector for the continuous measurement of the activity concentrations of alpha emitters in the culture media of in vitro cell suspension irradiation systems. The probe detector consists of a pen-size body housing a small silicon surface-barrier detector with a Mylar window. Theoretical calculations were performed to study the dependence of the alpha-energy spectrum on 1) the thickness of the Mylar barrier; 2) the Mylar-detector distance; and 3) the size of the detector window. These design parameters were selected by taking a compromise between the counting efficiency, the integrity of the detector, and its required range of application. The probe detector was tested using both chelated and unchelated 212Bi and 212Pb standard solutions; plate-out of these radionuclides on the Mylar barrier was observed for unchelated solutions. Alpha energy spectra were analyzed using a total integration technique. The measured activity concentrations and the calibrated values agree to within 4% for the chelated 212Bi and to within 6% for the unchelated 212Bi. The alpha probe detector can be used throughout an entire exposure time period to determine the total dose received by suspended cells, or at different time intervals to determine the dose rate in real time.

Alpha Particles↗

Noninvasive measurement of blood flow and extraction fraction.

We describe the theory of a technique for the noninvasive measurement of organ blood flow which is based on the principle of fractionation of cardiac output and is applicable with any recirculating gamma emitting tracer. The technique effectively determines the count rate that would be recorded over the organ if the tracer behaved like radiolabelled microspheres and was completely trapped in the organ's vascular bed on first pass. After correction for organ depth, the estimated first pass activity plateau, expressed as a fraction of the injected dose, is equal to the organ's fraction of the cardiac output (CO). By extending the theory, organ extraction fraction of extractable tracers or mean transit time of nonextractable tracers can be measured. Renal blood flow (RBF) to the normal left kidney in 18 subjects without evidence of renal disease was estimated by the technique to be 10.4% CO (S.D. 1.2), for the right kidney 9.0% CO(S.D. 1.1), and for both, 19.4% CO (S.D. 1.5). In a separate series of eight patients, RBF values were highly reproducible when obtained on successive days (standard deviation of change: 0.67% CO for the left kidney, 0.63% CO for the right and 0.78% CO for both). The extraction fraction of DTPA (filtration fraction) was 8.5% (1.7) in the left kidney and 9.7% (2.1) in the right kidney. This parameter was less reproducible than RBF, with standard deviations of the changes equal to 1.17% for the left kidney and 1.31% for the right. Stable, well-functioning, renal allografts in children had a mean blood flow of 20.6% CO (S.D. 3.7). Transplant blood flow in eight patients with rejection was less than 5.2% CO. In patients without splenomegaly, splenic blood flow, splenic extraction fraction of sulphur colloid and splenic red cell mean transit time were 4.3% CO (S.D. 0.9), 41% (S.D. 13.8), and 36.9 s (S.D. 4.6), respectively.

Blood Circulation↗

Kinetic Monte Carlo simulations with minimal searching.

Kinetic Monte Carlo (KMC) simulations are used to simulate epitaxial crystal growth. Presently, the fastest reported methods use binary trees to search through a list of rates in O(log(2) M) time, where M is the number of rates. These methods are applicable to an arbitrary set of rates, but typical KMC bond-counting schemes involve only a finite set of distinct rates. This allows one to construct a faster list-based algorithm with a computation time that is essentially independent of M. It is found that this algorithm typically reduces computation time by between 30% and 50% for typical simulations, with this factor increasing for larger simulations.

Journal Article↗

Efficient migration of complex off-line computer vision software to real-time system implementation on generic computer hardware.

This paper addresses the problem of migrating large and complex computer vision code bases that have been developed off-line, into efficient real-time implementations avoiding the need for rewriting the software, and the associated costs. Creative linking strategies based on Linux loadable kernel modules are presented to create a simultaneous realization of real-time and off-line frame rate computer vision systems from a single code base. In this approach, systemic predictability is achieved by inserting time-critical components of a user-level executable directly into the kernel as a virtual device driver. This effectively emulates a single process space model that is nonpreemptable, nonpageable, and that has direct access to a powerful set of system-level services. This overall approach is shown to provide the basis for building a predictable frame-rate vision system using commercial off-the-shelf hardware and a standard uniprocessor Linux operating system. Experiments on a frame-rate vision system designed for computer-assisted laser retinal surgery show that this method reduces the variance of observed per-frame central processing unit cycle counts by two orders of magnitude. The conclusion is that when predictable application algorithms are used, it is possible to efficiently migrate to a predictable frame-rate computer vision system.

Algorithms↗

Study of the efficacy of antiseptic handrub lotions with hand washing machines.

The effectiveness of quick handwashing in regards to four alcohol-based handrub lotions, including two products already on the market and our original lotions 1, 2, with two different kinds of hand washing machines was studied in vivo. We also tried to evaluate the efficacy of the four lotions in vitro. The in vivo testing of each lotion, including ethanol, was evaluated using two hand washing methods, first with and then without a previous soap wash. Computerized image analysis was used to calculate the bacterial count on the hand surface. The results showed that a 3 second application of the original lotion 2 (0.5% chlorhexidine in a 77% ethyl alcohol solution) preceded by a soap wash eradicated 89.3% of the bacteria on the hands; this was the highest reduction rate of all four detergents. AROKULIN-E (67.9% ethyl alcohol solution) without a prior handwashing produced the lowest reduction, 49.1% against hand surface bacteria. These results were compatible with those in vitro. It seems that an alcohol-based solution containing an effective antimicrobial detergent preceded by a soap wash is necessary to acceptably reduce hand surface bacteria.

Bacteria↗

Quantitative comparison of the transient rescue effects of neurotrophic factors on axotomized motoneurons in vivo.

A reproducible neuronal degeneration induced by nerve lesion in neonatal rats or mice provides a convenient in vivo assay for testing the survival-promoting activity of putative growth factors on motoneurons. The goal of this study was to compare the rescue effects of the four known neurotrophins [nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4 (NT-4)] and two of the cytokines [ciliary neurotrophic factor (CNTF) and leukaemia inhibitory factor (LIF)] in one particular experimental model of spinal motoneuron degeneration at two different survival times. The sciatic nerve was cut in neonatal rats and the factors were applied onto the nerve stump; bovine serum albumin was used in controls. Simultaneous application of the retrograde tracer fluoro-gold made it possible to count motoneurons specifically in the sciatic pool. One week after lesion, the neurotrophins BDNF, NT-3 and NT-4, but not NGF, equally enhanced motoneuron survival compared to controls; their effects were significantly better than those of the cytokines. However, the rescue from cell death was only transitory because a great number of the motoneurons died during the second week after nerve lesion. Additional BDNF and/or CNTF supplied by repeated subcutaneous injections (1 mg/ml) over 2 weeks could not prevent this delayed motoneuron loss. These results suggest that still other factors or alternative routes of administration may be required for permanent rescue of the lesioned immature motoneurons.

Animals↗

Changes in the number, size, and shape of synaptic vesicles in an experimental, projected cortical epileptic focus in the rat.

A projected epileptic focus was induced in the brains of experimental rats by applying penicillin G sodium to the contralateral hemisphere. Synapses of type I (after Gray) in the second cortical layer of the mirror area were analyzed morphometrically in the electron microscope 30 min after application of the epileptogenic drug. The number of agranular synaptic vesicles was counted per 0.01 micron2 over the active zone of the presynaptic membrane in an area 0.1 micron wide adjacent to the active zone and in an area of the same width 0.1 micron distant from the active zone. A statistically significant increase in the number of synaptic vesicles was found in the experimental animals as compared with the controls. The number of synaptic vesicles per 0.01 micron2 area was evaluated along the inactive presynaptic membrane, both in the area adjacent to the active zone of the presynaptic membrane and in other areas at successively greater distances from the active zone. The counts in some of these latter areas were statistically significantly higher in the experimental animals. The size and shape of synaptic vesicles were evaluated in three zones, each 0.1 micron wide, parallel to the membrane of the synaptic cleft. Zone I was directly adjacent to the membrane of the synaptic cleft; zone II was 0.1 micron distant; and zone III 0.1 micron distant from the membrane. The results show that the synaptic vesicles of both the experimental and control animals are significantly smaller in zone I than in the more distant zones. This trend is more marked in the controls than in the experimental animals. Histograms of vesical areas of zones II and III in the controls point to two populations of vesicles. The volumes of vesicles are approximately 10% greater in zone I of experimental animals than in the controls. The elongation of the vesicles in the three zones does not differ statistically either in controls or in experimental animals. Experimental animals, however, have statistically more rounded vesicles than the controls. Assuming that the synpatic vesicles contain the transmitters, our results might contribute to a better understanding of the increased excitability of the epileptic focus area.

Animals↗

Effect of insecticidal ear tags on populations of lice (Damalinia ovis) infesting sheep.

Polymer matrix ear tags containing 13.7% w/w tetrachlorvinphos or 8.5% w/w cypermethrin were applied to Merino wethers infested with lice and carrying two months wool. The cypermethrin tags reduced louse numbers by a maximum of 89% in comparison to controls at 16 weeks after treatment and by 85% at the conclusion of the experiment 38 weeks after application. Lice were not eradicated from any sheep. The number of lice counted on sheep treated with tetrachlorvinphos-impregnated tags was not significantly different from controls.

Analysis of Variance↗

Scanning-slit photon counting x-ray imaging system using a microchannel plate detector.

An experimental prototype of a novel photon counting x-ray imaging system was evaluated. This system is based on an "edge-on" microchannel plate (MCP) detector and utilizes scanning slit imaging configuration. The detector is capable of photon counting, direct conversion, high spatial resolution, controllable physical charge amplification, quantum limited and scatter free operation. The detector provides a 60 mm wide field of view (FOV) and its count rate is 200 kHz for the entire FOV. The count rate of the current system is limited by the position encoding electronics, which has a single input for all events from the entire detector, and incorporates a single channel ADC with 1 micros conversion time. It is shown that the count rate can potentially be improved to clinically acceptable levels using multichannel application specific integrated circuit (ASIC) electronics and multi-slit image acquisition geometry. For a typical acquisition time used in this study, the image noise was measured to be less than the typically acceptable noise level for medical x-ray imaging. It is anticipated that the noise level will be also low after the implementation of the ASIC electronics. The quantum efficiency of the detector was measured to be 40%-56% for an energy range of 50-90 kVp for MCPs used in this study and can be improved to > 80% using MCPs with the optimized parameters. Images of resolution and anthropomorphic phantoms were acquired at an x-ray tube voltage of 50 kVp. The value of contrast transfer function for the detector was measured to be 0.5 at a spatial frequency of 5 lp/mm. The intrinsic spatial resolution of the system is 28 microm FWHM and was limited by the accuracy of the time-to-digital conversion of the position encoding electronics. Given the advantages of the edge-on MCP detector such as direct conversion and physical charge amplification, it can potentially be applied to mammography and chest radiography.

Equipment Design↗

Extension of x-ray imaging linear systems analysis to detectors with energy discrimination capability.

A figure of merit, the broad-spectrum generalized detective quantum efficiency, which describes the performance of digital detectors designed for broad-spectrum x-ray imaging is derived from linear response theory. This measure of the imaging efficacy of an x-ray sensor is obtained when detector contrast modulation in the domain of x-ray energy is introduced in the Fourier-based analysis of digital systems. A method is proposed to scale existing figures of merit according to the energy-dependent response of the detector and the spectral shape of the x-ray beam. The new figure of merit obtained with this method provides an extended description of system performance when comparing energy-integrating, single-photon counting, and future energy-sensitive x-ray imaging sensors. The applicability of this linear system analysis is restricted to the tasks of low-contrast object detection in radiography. The method for scaling the figure of merit to take into consideration broad-spectrum conditions is applied to mammography for future energy-dependent detectors. An approximation valid in the typical mammographic x-ray energy range is used to calculate the broad-spectrum generalized detective quantum efficiency at zero spatial frequency, for several mammographic x-ray spectra. X-ray energy weighting in mammography is investigated in the context of simulated tumors and microcalcifications detection by comparing this figure of merit, calculated for different detector technologies, under ideal imaging conditions, at zero spatial frequency.

Algorithms↗

Novel real-time monitoring system for human cytomegalovirus-infected cells in vitro that uses a green fluorescent protein-PML-expressing cell line.

Promyelocytic leukemia (PML) bodies are discrete nuclear foci that are intimately associated with many DNA viruses. In human cytomegalovirus (HCMV) infection, the IE1 (for "immediate-early 1") protein has a marked effect on PML bodies via de-SUMOylation of PML protein. Here, we report a novel real-time monitoring system for HCMV-infected cells using a newly established cell line (SE/15) that stably expresses green fluorescent protein (GFP)-PML protein. In SE/15 cells, HCMV infection causes specific and efficient dispersion of GFP-PML bodies in an IE1-dependent manner, allowing the infected cells to be monitored by fluorescence microscopy without immunostaining. Since a specific change in the detergent solubility of GFP-PML occurs upon infection, the infected cells can be quantified by GFP fluorescence measurement after extraction. With this assay, the inhibitory effects of heparin and neutralizing antibodies were determined in small-scale cultures, indicating its usefulness for screening inhibitory reagents for laboratory virus strains. Furthermore, we established a sensitive imaging assay by counting the number of nuclei containing dispersed GFP-PML, which is applicable for titration of slow-growing clinical isolates. In all strains tested, the virus titers estimated by the GFP-PML imaging assay were well correlated with the plaque-forming cell numbers determined in human embryonic lung cells. Coculture of SE/15 cells and HCMV-infected fibroblasts permitted a rapid and reliable method for estimating the 50% inhibitory concentration values of drugs for clinical isolates in susceptibility testing. Taken together, these results demonstrate the development of a rapid, sensitive, quantitative, and specific detection system for HCMV-infected cells involving a simple procedure that can be used for titration of low-titer clinical isolates.

Anticoagulants↗

Quantitative study of nodulation competitiveness in Rhizobium strains.

We compared the nodulation competitiveness of three strains of Rhizobium leguminosarum by counting the number of nodules formed on faba bean plants after the application at sowing time of different concentrations of the strains to soils already containing Rhizobium strains of the same species. A relationship of type y = ax was found to exist between the ratio of the nodules formed by the applied inoculum strain to the nodules formed by the soil strains and the ratio of Rhizobium cells in the inoculum to the cells in the soil. This relationship was also confirmed in another competition experiment in which two R. meliloti strains of identical competitiveness were mixed in various proportions. The relationship can also be applied to the majority of results reported in the literature. Should it prove to be more widely applicable, it could be used to estimate the relative competitiveness of Rhizobium strains and thus predict the performance of an inoculum in a given soil.

Journal Article↗

Megacolon in rats.

Four months of sigmoid colon intrinsic denervation of young male Wistar rats resulted in typical megacolon. Average 46.0% denervation of the Auerbach plexus was obtained by serosal application of benzalkonium chloride (0.2% v/v). Denervation was assessed by ganglion cell counts in a 480-nm extension of the treated segment. Increased thickness of the megacolon wall was due to muscle hypertrophy (+117.4%) and mucosal hyperplasia (+53.9%) as shown by morphometry. The potential usefulness of this experimental model is emphasized.

Animals↗

The need to quantify right-to-left shunt in acute ischemic stroke: a case-control study.

BACKGROUND AND PURPOSE: Although right-to-left shunt (RLSh) has been reported to be significantly more frequent in young stroke patients with cryptogenic stroke, its relevance in a nonselected population of acute ischemic stroke is not well known. The aim of this study was to determine the importance of the RLSh magnitude as a risk factor for stroke in nonselected patients. METHODS: Two hundred eight patients hospitalized consecutively with transient ischemic attack or acute cerebral infarction and 100 healthy control subjects were studied. Transcranial Doppler ultrasonography (TCD) was performed in both middle cerebral arteries (MCAs) after intravenous application of agitated saline solution. The magnitude of RLSh was quantified by counting the number of signals in 1 MCA during a Valsalva maneuver. RLSh was classified as "no shunt," "small" (< 10 signals), and "large" (> 10 signals), with the latter including the "shower" (> 25 signals) and "curtain" (uncountable signals) patterns. Extensive investigations, including contrast transesophageal echocardiography, were carried out on patients diagnosed as suffering from stroke of an uncertain etiology. The importance of RLSh for stroke was assessed by logistic regression analysis. RESULTS: Contrast TCD detected a large RLSh in 40 (19.7%) patients and in 21 (21%) control subjects, all with cardiac RLSh characteristics. A large RLSh was present in 4.7% of atherothrombotic strokes, 10.5% of cardioembolic strokes, 15.4% of lacunar strokes, and 45.3% of cryptogenic strokes (P<0.001). Although the overall frequency of RLSh was not significantly different between patients and control subjects, the detection of curtain or shower patterns by contrast TCD was associated with a higher risk of stroke (odds ratio, 3.5; 95% confidence interval, 1.29 to 9.87), particularly with cryptogenic stroke (odds ratio, 12.4; 95% confidence interval, 4.08 to 38.09) after adjustment for concomitant vascular risk factors. CONCLUSIONS: It is essential to quantify RLSh by contrast TCD during the Valsalva maneuver given that only those with shower and curtain patterns are associated with a higher risk of ischemic stroke in a nonselected population.

Acute Disease↗