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Biomedical subjects

P Munro

Publications and source records attributed to P Munro.

At least 19 recordsLinked to original sources

Emulsion strategies in the microencapsulation of cells: pathways to thin coherent membranes.

Microencapsulation of cell spheroids in an immunoselective, highly biocompatible, biomembrane offers a way to create viable implantation options in the treatment of insulin-dependent diabetes mellitus (IDDM). Traditionally the encapsulation process has been achieved through the injection/extrusion of alginate/cell mixtures into a calcium chloride solution to produce calcium alginate capsules around the cells. A novel alternative is explored here through a procedure using an emulsion process to produce thin adherent calcium alginate membranes around cell spheroids. In this study, a thorough investigation has been used to establish the emulsion process parameters that are critical to the formation of a coherent alginate coat both on a model spheroid system and subsequently on cell spheroids. Optical and fluorescence microscopy are used to assess the morphology and coherence of the calcium alginate/poly-L-ornithine/alginate (APA) capsules produced.

Alginates↗

Bacterial toxins activating Rho GTPases.

The CNF1 toxin is produced by some uropathogenic (UPECs) andmeningitis-causing Escherichia coli strains. It belongs to a large family of bacterial virulence factors and toxins modifying cellular regulators of the actin cytoskeleton, namely the Rho GTPases. CNF1 autonomously enters the host cell cytosol, where it catalyzes the constitutive activation of Rho GTPases by deamidation. This activation is, however, attenuated because of activated Rho protein ubiquitin-mediated proteasomal degradation. Both Rho protein activation and deactivation confer phagocytic properties on epithelial and endothelial cells, as well as epithelial cell motility and cell-cell junction dynamics. Transcriptome analysis using DNA microarray revealed that endothelial cells respond to high doses of CNF1 by launching a genetic program of host alarm. This host cell reaction to CNF1 intoxication also indicates that degradation of activated Rho proteins by the proteasome may lead to a lowering of the threshold of the intoxicated cell inflammatory response. These results are consistent with growing evidence that Rho proteins control the cell inflammatory responses. It is tempting to assume that Rho deregulation may participate in various immunological disorders also involved in cancer.

Animals↗

Monte Carlo simulation of the image formation process in portal imaging.

We have written Monte Carlo programs to simulate the formation of radiological images. Our code is used to propagate a simulated x-ray fluence through each component of an existing video-based portal imaging system. This simulated fluence consists of a 512 x 512 pixel image containing both contrast-detail patterns as well as checker patterns to assess spatial resolution of the simulated portal imager. All of the components of the portal imaging system were modeled as a cascade of eight linear stages. Using this code, one can assess the visual impact of changing components in the imaging chain by changing the appropriate probability density function. Virtual experiments were performed to assess the visual impact of replacing the lens and TV camera by an amorphous silicon array, and the effect of scattered radiation on portal images.

Computer Simulation↗

Kilovision: thermal modeling of a kilovoltage x-ray source integrated into a medical linear accelerator.

The thermal and thermo-mechanical (fatigue) properties of a stationary-anode kilovoltage x-ray source that can be integrated into the head of a medical linear accelerator have been modeled. A finite element program has been used to model two new target designs. The first design makes minor modifications to the existing target assembly of a Varian medical linear accelerator, while the second design adds an additional cooling tube, changes the target angle, and uses a tungsten-rhenium alloy rather than tungsten as the kilovoltage target material. The thermal calculations have been used to generate cyclic stress/strain values from which estimates of fatigue in the target designs have been made. Both kilovoltage and megavoltage operation have been studied. Analysis of the megavoltage operation shows that there are only small differences in the thermal and fatigue characteristics after the target assembly is modified to include a kilovoltage target. Thus, megavoltage operation should not be compromised. The first kilovoltage target design can handle a 900 W heat load (e.g., 120 kVp, 7.5 mA, 2 x 2 mm2 source size); the heat load being limited by the temperature at the surface of the cooling tubes and mechanical fatigue at the surface of the target. The second design can handle a 1250 W heat load (e.g., 120 kVp, approximately 10.4 mA, 2 x 2 mm2 source size). Our calculations show that installation of a kilovoltage x-ray target is practical from thermal and thermo-mechanical perspectives.

Computer Simulation↗

High sensitivity of mouse neuronal cells to tetanus toxin requires a GPI-anchored protein.

Tetanus neurotoxin (TeNT) produced by Clostridium tetani specifically cleaves VAMP/synaptobrevin (VAMP) in central neurons, thereby causing inhibition of neurotransmitter release and ensuing spastic paralysis. Although polysialogangliosides act as components of the neurotoxin binding sites on neurons, evidence has accumulated indicating that a protein moiety is implicated as a receptor of TeNT. We have observed that treatment of cultured mouse neuronal cells with the phosphatidylinositol-specific phospholipase C (PIPLC) inhibited TeNT-induced cleavage of VAMP. Also, we have shown that the blocking effects of TeNT on neuroexocytosis can be prevented by incubation of Purkinje cell preparation with PIPLC. In addition, treatment of cultured mouse neuronal cells with cholesterol sequestrating agents such as nystatin and filipin, which disrupt clustering of GPI-anchored proteins in lipid rafts, prevented intraneuronal VAMP cleavage by TeNT. Our results demonstrate that high sensitivity of neurons to TeNT requires rafts and one or more GPI-anchored protein(s) which act(s) as a pivotal receptor for the neurotoxin.

Animals↗

Technique charts for EC film: direct optical measurements to account for the effects of X-ray scatter.

PURPOSE: To develop a method of measuring technique charts for enhanced contrast (EC) film, to demonstrate how X-ray scatter changes the response of EC film, and to generate technique charts for general use. METHODS AND MATERIALS: We have developed a "digital cassette"-consisting of a metal plate/phosphor screen, a light guide, a photodiode sensor, and an electrometer-that can be used to measure the light generated in the phosphor screen of the film cassette. In turn, these measurements can be used to generate technique charts for EC film. The digital cassette has been used to measure technique charts for 4-MV and 6-MV X-ray beams for a variety of different phantom thicknesses, field sizes, and phantom-to-cassette air gaps. RESULTS AND DISCUSSION: We have observed that the signals generated in an ionization chamber located 9.4 cm behind a 30-cm-thick water-equivalent phantom increase by a factor of 1.9 when the field size is increased from 4 x 4 cm(2) to 40 x 40 cm(2) when irradiated by a 6-MV X-ray beam. However, the change in EC film response is a factor of 3.5 under the same conditions. Irradiations to optimally expose the EC film predicted by the digital cassette differ by up to 82% compared to those predicted by ion chamber measurements. Nevertheless, the technique charts measured using the digital cassette predict the response of the EC film to +/- 0.2 optical density. The overresponse of the EC film is most likely due to low-energy scattered photons, which interact with the high atomic number (Z = 64) phosphor screen of the enhanced contrast localization cassette. Therefore, simple solutions, such as placing a high atomic number material above the enhanced contrast localization cassette, can reduce this contribution by scattered photons to the signal generated in the cassettes. CONCLUSIONS: We have developed a digital cassette that can make more accurate measurements of the technique charts for EC films. Our measurements show that under some conditions, X-ray scatter can generate a large fraction of the signals recorded by the EC film. Technique charts have been generated at 4 MV and 6 MV, and these charts should have universal applicability.

Film Dosimetry↗

Epidemic dystonia in Cork.

We describe a case series of five patients with unusual presentations of acute dystonia seen over a 2-week period. Haloperidol, prescribed and allegedly purchased 'on the street', was thought to be a possible cause in each case. Bizarre clinical presentations and delayed onset of symptoms may make diagnosis difficult. A high index of suspicion combined with a careful drug history is essential.

Adolescent↗

Clinical use of electronic portal imaging: report of AAPM Radiation Therapy Committee Task Group 58.

AAPM Task Group 58 was created to provide materials to help the medical physicist and colleagues succeed in the clinical implementation of electronic portal imaging devices (EPIDs) in radiation oncology. This complex technology has matured over the past decade and is capable of being integrated into routine practice. However, the difficulties encountered during the specification, installation, and implementation process can be overwhelming. TG58 was charged with providing sufficient information to allow the users to overcome these difficulties and put EPIDs into routine clinical practice. In answering the charge, this report provides; comprehensive information about the physics and technology of currently available EPID systems; a detailed discussion of the steps required for successful clinical implementation, based on accumulated experience; a review of software tools available and clinical use protocols to enhance EPID utilization; and specific quality assurance requirements for initial and continuing clinical use of the systems. Specific recommendations are summarized to assist the reader with successful implementation and continuing use of an EPID.

Biophysical Phenomena↗

Deamidation of RhoA glutamine 63 by the Escherichia coli CNF1 toxin requires a short sequence of the GTPase switch 2 domain.

CNF1, a toxin produced by pathogenic Escherichia coli strains, deamidates the RhoA GTP-binding protein glutamine 63 and impairs RhoGAP-mediated GTP hydrolysis resulting in RhoA permanent activation. Using peptides derived from the RhoA sequence, we found that DTAGQEDYDRL (corresponding to RhoA 59-69 residues) was the minimum RhoA-derived peptide which could be deamidated in vitro by the CNF1 catalytic domain (CNF1-Cter). Site-directed mutagenesis outside the RhoA 59-69 sequence had no influence on glutamine 63 deamidation by CNF1-Cter. RhoA proteins with substitutions L57G, D65G, Y66G, or R70G were not affected in their ability to be deamidated by CNF1-Cter, whereas this was abolished by the R68G substitution. Arginine 68 is part of the DYDRL motif that is strictly conserved in Rho, Rac, and Cdc42 but not in other small GTP-binding proteins consistent with the observation that only Rho, Rac, and Cdc42 can be modified by CNF1.

Amino Acid Sequence↗

Subjectivity in interpretation of portal films.

PURPOSE: We have measured the variability in identifying geometric errors and the variability in clinical decision making when using portal films. METHODS AND MATERIALS: Eight observers (four radiation oncologists and four radiation therapists) viewed 40 film pairs from 40 different patients. All films, which were acquired using conventional simulator and portal film cassettes, were selected retrospectively from a large clinical database. The observers compared the simulator and portal films under standard conditions, identified the field placement errors, and decided whether adjustments in treatment were required. In addition, all films were digitized and the field placement errors were measured objectively using image registration software. RESULTS: There was much variability in identifying field placement errors and even more variability in the number of recommended adjustments. The field placement errors identified by the different observers differed by up to 50 mm for the same film pair. The number of adjustments of treatment or block position recommended by the observers also varied between 8 and 25 for the same set of films. The average field placement error, before correction, for AP lung, AP pelvis, and lateral pelvis films was 5.7 mm, 6.3 mm, and 8.9 mm, while the average error after correction (i.e., correcting all errors identified by the observers) was 5.5 mm, 4.9 mm, and 5.7 mm, respectively. Thus, for lateral pelvis films, where the initial errors were larger, the observers were able to make an improvement in patient setup. CONCLUSIONS: The results suggest that human observers have difficulty identifying field placement errors accurately when the errors are around 5 mm or smaller. Although there is some evidence that experience influenced the performance of the observers, the effect of experience is not large. In routine clinical environments, the use of visual inspection will detect large field placement errors. However, tools other than visual inspection will be required if field placement errors 5 mm or smaller are to be identified accurately.

Decision Making↗

Biofilm on scleral explants with and without clinical infection.

PURPOSE: Biofilm is a glycocalyx matrix secreted by microorganisms that confers protection against host defenses and antimicrobial treatment. Biofilms have been implicated in the persistence of scleral buckle infections. This study aimed to evaluate the incidence of biofilm growth on scleral explants and the relationship to explant infection. METHODS: Scleral explants were obtained following removal for infection or extrusion or during repeat surgery. Explants were fixed with rhuthenium red and examined by scanning electron microscopy to visualize the glycocalyx. RESULTS: A total of 28 explants were analyzed. Ten were removed because of either infection or extrusion and 18 were removed during repeat surgery. The mean time to removal of explants was 36 months in the infection/extrusion group and 12 months in the others. Biofilm was identified on five explants-two removed because of infection/extrusion and three for surgical indications. Bacterial elements were identified in all biofilms. CONCLUSIONS: Biofilm was identified on explants removed because of infection or exposure and on explants removed for technical reasons at repeat surgery. This implies that bacterial contamination and biofilm formation occur without exposure of the explant, probably due to inoculation at the time of initial surgery. Biofilms may contribute to the persistence of scleral explant infections but a causative role in buckle extrusion is unproved.

Biofilms↗

Toxins from anaerobic bacteria: specificity and molecular mechanisms of action.

Major advances have been made in the past five years in the identification of cellular targets of toxins produced by anaerobic bacteria. These targets include the vesicular membrane docking and fusion apparatus, the actin cytoskeleton, the signal transduction machinery and the cell membrane. The recent discovery that large clostridial toxins (Clostridium difficile A and B toxins, C. sordellii lethal and hemorrhagic toxins, and alpha C. novyi toxin) are monoglucosyltransferases, together with the establishment of the perfringolysin crystal structure, has led to new insights in the field of toxins from anaerobic bacteria.

Bacterial Toxins↗

An ultrastructural and systemic analysis of glycosaminoglycans in thyroid-associated ophthalmopathy.

PURPOSE: To determine the ultrastructural localisation of glycosaminoglycans (GAGs) in the extraocular muscles (EOMs) of patients with thyroid-associated ophthalmopathy (TAO) and to see whether the quantity and type of GAGs present in blood and urine are markers of the disease. METHODS: Biopsies of affected EOMs were taken and studied by transmission electron microscopy (TEM). These were either fixed conventional for TEM, or in 0.5% tannic acid and others for immunogold staining. Serum hyaluronan (HA) was measured using a radioimmunoassay in patients with TAO as well as control subjects, and urinary GAG levels assessed by photometric quantitation of hexuronic acid after reaction with carbazole. The excretion pattern of the urinary GAGs was determined by discontinuous electrophoresis. RESULTS: TEM showed that there is a marked expansion of the endomysial space in TAO EOM biopsies as compared with non-TAO strabismus specimens. This is caused by an increased number of collagen fibres, interspersed with a granular amorphous material surrounding striated collagen fibres shown to be hyaluronan by immunogold staining. In contrast, serum hyaluronan concentrations were similar in TAO and control patients, although there was a statistically significant difference in the urinary GAG excretion between the two groups of patients examined. By discontinuous electrophoresis, chondroitin sulphate and heparan sulphate were present in both patients and controls. CONCLUSION: GAGs and in particularly HA are present at the EOM level in patients with recently inactive TAO. However, serum levels of HA and urinary GAGs are not sensitive indicators for their presence within the EOMs.

Adult↗

A Monte Carlo study of verification imaging in high dose rate brachytherapy.

We have been evaluating the practicality of monitoring the position of an 192Ir source during high dose rate (HDR) brachytherapy treatments using x-ray fluoroscopy. The EGS4 Monte Carlo code has been used to simulate the interactions of 192Ir photons with the patient and the CsI phosphor of an x-ray image intensifier to predict what signals will be generated by these 192Ir photons. The calculations show that it is the 192Ir photons scattered within the patient that are mainly responsible for generating the spurious signals in the x-ray image intensifier that degrade image quality. The scattered 192Ir photons are distributed in the energy range (15-200 keV), which is markedly lower than the average energy of the primaries (360 keV), and therefore interact more efficiently with the CsI phosphor of the x-ray image intensifier. Experimental measurements support these observations, demonstrating that spurious signals produced by the 192Ir source become appreciably larger when the 192Ir source is located within a scattering object rather than air. For a 10 cm airgap, the signal-to-noise ratio (SNR) can decrease by factors ranging between 3 and 10 (no antiscatter grid), depending on the position of a 7 Ci 192Ir source inside a 30 cm thick water phantom. In typical clinical situations, a focused grid (Pb, 12:1, 40 lines/cm) can increase the SNR by about a factor of 2. Furthermore, the SNR rapidly increases with increasing airgap, such that a 20 cm airgap can be as effective as a 12:1 air interspaced grid in eliminating the spurious signals. Our results suggest that use of a high-current x-ray fluoroscopy technique, a large airgap, and a well-designed anti-scatter grid can make the fluoroscopic monitoring of source position in HDR brachytherapy feasible. This, in turn, can improve the quality assurance of such treatments.

Algorithms↗

X-ray quantum limited portal imaging using amorphous silicon flat-panel arrays.

We have measured the linearity, spatial resolution (MTF), noise (NPS), and signal-to-noise characteristics (DQE) of an electronic portal imaging device (EPID) based on an amorphous silicon flat-panel array. The array has a 128 x 128-pixel matrix and each pixel is 0.75 x 0.75 mm2 in dimension so the array covers an area of 96 x 96 mm2. The array acts like a large area light sensor and records the optical signals generated in a metal plate/phosphor screen x-ray detector when the detector is irradiated by a megavoltage x-ray beam. In addition, approximately 0.5% of the total signal is generated by nonoptical processes. The noise measurements show that the device is quantum noise limited with the noise power generated by the x-ray quanta being up to 100 times greater than the noise added by the external readout electronics and flat-panel light sensor itself. However, the flat-panel light sensor does reduce the spatial resolution (compared to a perfect optical sensor with infinitesimal pixel size) because of its moderate pixel size and because optical spread can occur in the transparent glues used to attach the phosphor screen to the flat-panel light sensor. The response of the sensor is very linear and does not suffer from the glare phenomenon associated with TV camera-based EPIDs--characteristics which suggest that the amorphous silicon EPID will be well suited to transit dosimetry. Nevertheless, some limitations need to be overcome before these devices can be used clinically. These include developing larger flat-panel light sensors, the elimination of "noisy" pixels with high dark signal, and improvements in the uniform sensitivity of the sensors. This last requirement is only needed for transit dosimetry applications where it would greatly simplify calibration of the device. In addition, an image acquisition scheme must be developed to eliminate artifacts created by the pulsed x-ray beam generated by linear accelerators. Despite these limitations, our studies suggest that the amorphous silicon EPIDs are very well suited to portal imaging.

Artifacts↗

Optimal phosphor thickness for portal imaging.

A theoretical approach known as quantum accounting diagram (QAD) analysis has been used to calculate the spatial-frequency-dependent detective quantum efficiency (DQE) of two portal imaging systems: one based on a video camera and another based on an amorphous silicon array. The spatial frequency-dependent DQEs have then been used to determine indices of displayed and perceived image quality. These indices are figures of merit that can be used to optimize the design of linear imaging systems. We have used this approach to determine which of eight phosphor screen thicknesses (ranging between 67 and 947 mg/cm2) is optimal for the two designs of portal imaging systems. The physical characteristics (i.e., detection efficiencies, gains, and MTFs) of each of the eight x-ray detectors have been measured and combined with the physical characteristics of the remaining components to calculate the theoretical DQEs. In turn, the DQEs have been used to calculate theoretical indices of displayed and perceived image quality for two types of objects: a pelvis object and a pointlike object. The maximal indices of displayed and perceived image quality were obtained with screen thickness ranging between 358 and 947 mg/cm2, depending upon the imaging system design and the object being imaged. Importantly, the results showed that there is no single optimal screen thickness. The optimal thickness depended upon imaging task (e.g., detecting large, low-contrast structures, or detecting edges and small structures). Nevertheless, the results showed that there were only modest improvements in the indices of image quality for phosphor screens thicker than 350-400 mg/cm2.

Biophysical Phenomena↗

A quantum accounting and detective quantum efficiency analysis for video-based portal imaging.

The quality of images generated with radiographic imaging systems can be degraded if an inadequate number of secondary quanta are used at any stage before production of the final image. A theoretical technique known as a "quantum accounting diagram" (QAD) analysis has been developed recently to predict the detective quantum efficiency (DQE) of an imaging system as a function of spatial frequency based on an analysis of the propagation of quanta. It is used to determine the "quantum sink" stage(s) (stages which degrade the DQE of an imaging system due to quantum noise caused by a finite number of quanta), and to suggest design improvements to maximize image quality. We have used this QAD analysis to evaluate a video-based portal imaging system to determine where changes in design will have the most benefit. The system consists of a thick phosphor layer bonded to a 1 mm thick copper plate which is viewed by a T.V. camera. The imaging system has been modeled as ten cascaded stages, including: (i) conversion of x-ray quanta to light quanta; (ii) collection of light by a lens; (iii) detection of light quanta by a T.V. camera; (iv) the various blurring processes involved with each component of the imaging system; and, (v) addition of noise from the T.V. camera. The theoretical DQE obtained with the QAD analysis is in excellent agreement with the experimental DQE determined from previously published data. It is shown that the DQE is degraded at low spatial frequencies (< 0.25 cycles/mm) by quantum sinks both in the number of detected x rays and the number of detected optical quanta. At higher spatial frequencies, the optical quantum sink becomes the limiting factor in image quality. The secondary quantum sinks can be prevented, up to a spatial frequency of 0.5 cycles/mm, by increasing the overall system gain by a factor of 9 or more, or by improving the modulation transfer function (MTF) of components in the optical chain.

Biophysical Phenomena↗

Evaluation of a high-density scintillating glass for portal imaging.

One of the main factors that limits the performance of T.V. camera-based portal imaging systems is the poor light-collection efficiency of the lens and T.V. camera. An x-ray detector that produces more light per incident x ray would help overcome this limitation. We have been evaluating a high-density (3.8 g/cm3), thick (12 mm) glass scintillator for its suitability as an x-ray detector for T.V. camera-based portal imaging systems. The light output and spatial resolution of the glass scintillator has been compared to that of a copper plate/phosphor screen detector using radiographic film and the T.V. camera of our portal imaging system. The film measurements show that the light output of the glass scintillator is 82% of that of the copper plate/phosphor screen, while the T.V. camera measurements show that this value is 48%. A theoretical model of light transport described in this paper suggests that this discrepancy is due to refraction at the glass-air interface. Our measurements of the modulation transfer function (MTF) show that the spatial resolution obtained with the glass scintillator is similar to that obtained with the copper plate phosphor screen. However, the spatial resolution obtained with the glass scintillator decreases as the angle of x-ray incidence increase; this decrease, which is not observed for the copper plate/phosphor screen detector, is due to the large thickness of the glass scintillator. Due to the limited light output and the variable spatial resolution, the transparent glass scintillator, in its current form, is not suitable for portal imaging.

Biophysical Phenomena↗