Gene expression via direct injection of plasmid vector into the thymus.
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Biomedical subjects
Publications and source records attributed to R Pastori.
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Digital radiography (DR) based on storage phosphor technology is progressively replacing conventional screen-film system radiographic techniques (CR). However, many questions about image quality and dose reduction are still open. Thus, since DR spatial resolution is always lower than that of high quality screen-film images and image noise is conversely higher, for general radiography examinations especially, the signal-to-noise ratio is always lower with DR than with CR at the same dose level. However, the wide dynamic range, the linear response of storage phosphor detectors and the automatic read-out control of the digital system, enable lower DR X-ray imaging efficiency to be overcome, producing a nearly perfect readout of the image data every time an exposure is made. Moreover, post-processing can make the detection of abnormalities easier. This is why in many ROC clinical accuracy studies DR performed equally or better than CR. Good quality DR images, similar to conventional ones at the same dose levels, are therefore achievable in chest, abdomen, bone and soft-tissues examinations. Dose evaluation protocols are strongly needed in DR since overexposures due to technical mistakes may be overlooked; moreover, doses can be adjusted to the peculiar diagnostic need. The S values shown on any hard-copy image can be used to this purpose, even though it must be pointed out that they do not express the actual mean dose to the detector.
To evaluate the diagnostic possibilities of Digital Radiography (DR) and of Conventional Radiography (CR) in the diagnosis of fine chest lesions, 100 patients with AIDS were examined with both techniques. The gold standards were 67Ga scintigraphy and bronchial washing in 100 patients and Computed Tomography (CT) in 50 patients. The data obtained from DR and CR examinations were separately evaluated by 4 readers and subsequently analyzed by ROC curves with Standard Error calculation and by comparing the areas below the characteristic curves with Student's paired t-test (p less than or equal to 0.05). Statistical analysis showed DR and CR to be of equivalent diagnostic value in the study of interstitial lung diseases.
To date, the skeletal imaging capabilities of digital radiography with storage phosphors have been poorly investigated, and the diagnostic accuracy of this technique has not been thoroughly assessed. To evaluate the performance of storage phosphor digital radiography we compared 66 conventional and 66 digital radiographs of small abnormalities of the extremities (fractures, erosions, calcifications). Conventional images were obtained with a low-speed screen-film system while digital ones were acquired with high-resolution (5 lp/mm max) phosphors and laser-printed on a 8" x 10" film. Two experienced radiologists defined the gold standard (389 abnormalities) and four radiologists scored the findings (1,556 observations) on a five-point discrete scale. ROC analysis indicated film and storage radiography to be equally effective in the overall detection of abnormalities. No difference was found in the individual performances of the four readers, in the site subclasses (wrist, hand), and in the specific detection of fractures and erosions. Digital radiography proved to be superior to conventional radiography in the detection of calcifications in all sites and particularly in the wrist (p less than 0.05). Storage phosphor radiography may replace conventional radiography of the extremities without causing any significant decrease in diagnostic accuracy.
Partial activation of Mucor rouxii cAMP-dependent protein kinase by cAMP was obtained when kemptide was used as substrate, but complete activation was attained with cAMP plus protamine or histone. Full activation could not be achieved by increasing kemptide or cAMP concentration. Complete activation by cAMP could be obtained by addition of 10 microM polylysine, 10 microM lysine-rich histone or 0.5 mM spermine plus spermidine. The degree of stimulation could be up to 5-fold, depending on the amount of enzyme in the assay. The same concentrations of polycations increased 1.5-2.3-fold the Vmax of kemptide phosphorylation by the free catalytic subunits of both Mucor and bovine heart protein kinases; 10 microM polyarginine inhibited completely the activity of both enzymes.
A digital radiographic system using storage phosphor detectors was employed in order to verify radiotherapy treatments with high energy photon beams (60Co-18 MV X-rays). The wide range linearity of the detectors and also the possibility of digital image post-processing allowed portal films to be obtained with sufficient contrast in all treatment techniques, particularly in the case of mantle fields. The use of digital radiography may become an easy and valuable procedure for therapy verification.
The polymeric structure of the cyclic AMP-dependent protein kinase (E.C.2.7.1.37) from the dimorphic fungus Mucor rouxii was analyzed through studies of gel filtration and sucrose gradient centrifugation of the holoenzyme and its subunits and by photoaffinity labeling of the regulatory subunit. It was demonstrated that it is a tetramer composed by two regulatory subunits (R) of mol. wt. 75 000 and two catalytic subunits (C) of mol. wt. 41 000 forming a holoenzyme R2C2 of mol. wt. 242 000. Frictional coefficients of 1.55 and 1.62 for the holoenzyme and for the regulatory dimer, respectively, indicate a significant degree of dimensional asymmetry in both molecules. A procedure for the purification of the catalytic subunit of the kinase is presented. The holoenzyme could be bound to a cyclic AMP-agarose column and the catalytic subunit could be eluted by 0.5 M NaCl, well resolved from the bulk of protein. This particular behaviour of the holoenzyme in cyclic AMP-agarose chromatography allowed the inclusion of this step in the purification of the catalytic subunit and corroborated that the holoenzyme was not dissociated by cyclic AMP alone. The isolated catalytic subunit displays Michaelis-Menten behavior towards kemptide, protamine and histone and is inhibited by sulfhydryl reagents, indicating that the molecule has at least one cysteine residue essential for enzyme activity. The catalytic activity of the isolated C subunit is inactivated by the mammalian protein kinase inhibitor, and is inhibited by the regulatory subunit from homologous and heterologous sources. In general, the properties of the catalytic subunit suggest a structural similarity between Mucor and mammalian C subunits.
Cyclic AMP binding to Mucor rouxii protein kinase holoenzyme and free regulatory subunit was measured by the classical membrane filtration technique and by equilibrium dialysis. The results obtained demonstrate that the filtration method can be used without loss of any cyclic AMP binding site. Both methods unambiguously demonstrate that the number of molecules of cyclic AMP bound to the holoenzyme are half of those bound to the regulatory subunit. This result suggests that unshielding of new cyclic AMP binding sites occurs upon dissociation of the ternary complex holoenzyme-cyclic AMP.
The clinical utility was evaluated of a computed radiographic system in urography. The system (FCR 101, Philips Medical Systems, Inc., Shelton, CT) is based on a photo-stimulatable phosphor screen (imaging plate) for X-ray image detection and storage. The X-ray information recorded on the imaging plate is converted into digital from and processed by means of a computer. After processing is completed, the digitized image is reversed back to analogic signals, which modulate the intensity of a laser beam scanning the image on a single-emulsion film (Fuji CR 633). Two hundred IVP's were obtained in four groups, of 50 patients each, with normal azotemic values by rapid infusion of a low osmolality contrast medium (iopamidol 150 mgI/ml). While conventional radiographs were performed on the first group of patients with the injection of 0.6 gI/kg body weight of contrast medium, digital examinations were carried out, in the remaining three groups, with the injection of 0.6, 0.3 and 0.12 gI/kg, respectively. The digital images were processed with the "Abdomen-routine" program. A specific algorithm was implemented in order to reduce the excessive contrast resolution of the bladder, which is due to the characteristics of the nonionic contrast medium and enhanced by the reading program. The image details were evaluated by two observers and then statistically analyzed with nonparametric tests. Statistical analysis did not show any difference in the quality of digital and screen-film images. Image processing improved some inadequate images, by reducing the contrast resolution of the bladder, and allowed a better detection of some details. Low doses (0.3 gI/kg) of a low osmolality (150 mgI/kg) contrast medium were enough to obtain good images. Another biological advantage was obtained by a consistent radiation dose reduction (about 40%).
Aim of this work is to evaluate the diagnostic accuracy of different modalities of digital radiology, compared with conventional radiology, in the detection of lung nodules. Forty images of an anthropomorphic chest phantom, on which simulated nodules had been previously set, were analyzed by 6 radiologists with 4 different viewing modalities: a) conventional radiography; b) photostimulable plates digital radiography; c) photostimulable plates digital image on dedicated monitor; d) conventional radiography digitized with Film Laser Digitizer and viewed on Display Workstation. For each image the radiologists had to transfer on a sketch the site of the detected nodules giving an opinion on their detectability (uncertain-presumable-probable-certain presence) with a 1 to 4 score. The statistical analysis of the results was made using modified ROC curves. No statistically significant difference was found between the areas under a) and c) ROC curves (p = 0.262). A statistically significant difference resulted between the areas under a) and d) ROC curves (p < 0.05) and, particularly, between the a) and b) areas (p < 0.001), in favor of a).
Conventional and digital mammographic images obtained with storage phosphors were compared. The digital images were acquired with high-resolution 3rd-generation (HR III) screens and specifically adapted algorithms. The experience was made both on a phantom and in vivo. The phantom study was carried out by comparing conventional with digital radiographs acquired with: a) same kV (28) and same mAs; b) same kV (24) and same mAs; c) 28 kV with 30% mAs reduction, in digital images only. The results obtained upon counting the amount of recognizable details per image demonstrated slight loss of information in digital radiographs only when mAs was reduced by 30%. Two hundred patients were studied; they were divided into four groups according to the type of breast (medium inherent contrast or solid breast) and to exposure factors. After conventional mammography, an additional digital radiograph was performed using one of the three techniques previously employed on the phantom. Separately, 10 ductogalactographies and 10 magnification radiographs of microcalcifications were compared; the same exposure factors were used in these images too. The radiographs were viewed by three expert mammographic radiologists; contrast quality and spatial resolution were investigated and a score was given to each image on a 3-grade scale (insufficient, sufficient, good). The comparison of the mean values obtained showed higher contrast and better enhancement of nodular lesions on digital images, on which, however, the depiction of microcalcifications was worse than on conventional radiographs, especially with lower radiation doses, even though the number of detected microcalcifications was the same.