Estimating radiation doses from multidetector CT using Monte Carlo simulations: effects of different size voxelized patient models on magnitudes of organ and effective dose.

The purpose of this work is to examine the effects of patient size on radiation dose from CT scans. To perform these investigations, we used Monte Carlo simulation methods with detailed models of both patients and multidetector computed tomography (MDCT) scanners. A family of three-dimensional, voxelized patient models previously developed and validated by the GSF was implemented as input files using the Monte Carlo code MCNPX. These patient models represent a range of patient sizes and ages (8 weeks to 48 years) and have all radiosensitive organs previously identified and segmented, allowing the estimation of dose to any individual organ and calculation of patient effective dose. To estimate radiation dose, every voxel in each patient model was assigned both a specific organ index number and an elemental composition and mass density. Simulated CT scans of each voxelized patient model were performed using a previously developed MDCT source model that includes scanner specific spectra, including bowtie filter, scanner geometry and helical source path. The scan simulations in this work include a whole-body scan protocol and a thoracic CT scan protocol, each performed with fixed tube current. The whole-body scan simulation yielded a predictable decrease in effective dose as a function of increasing patient weight. Results from analysis of individual organs demonstrated similar trends, but with some individual variations. A comparison with a conventional dose estimation method using the ImPACT spreadsheet yielded an effective dose of 0.14 mSv mAs(-1) for the whole-body scan. This result is lower than the simulations on the voxelized model designated 'Irene' (0.15 mSv mAs(-1)) and higher than the models 'Donna' and 'Golem' (0.12 mSv mAs(-1)). For the thoracic scan protocol, the ImPACT spreadsheet estimates an effective dose of 0.037 mSv mAs(-1), which falls between the calculated values for Irene (0.042 mSv mAs(-1)) and Donna (0.031 mSv mAs(-1)) and is higher relative to Golem (0.025 mSv mAs(-1)). This work demonstrates the ability to estimate both individual organ and effective doses from any arbitrary CT scan protocol on individual patient-based models and to provide estimates of the effect of patient size on these dose metrics.

Proline-dependent oligomerization with arm exchange.

BACKGROUND: Oligomerization is often necessary for protein activity or regulation and its efficiency is fundamental for the cell. The quaternary structure of a large number of oligomers consists of protomers tightly anchored to each other by exchanged arms or swapped domains. However, nothing is known about how the arms can be kept in a favourable conformation before such an oligomerization. RESULTS: Upon examination of such quaternary structures, we observe an extremely frequent occurrence of proline residues at the point where the arm leaves the protomer. Sequence alignment and site-directed mutagenesis confirm the importance of these prolines. The conservation of these residues at the hinge regions can be explained by the constraints that they impose on polypeptide conformation and dynamics: by rigidifying the mainchain, prolines favour extended conformations of arms thus favouring oligomerization, and may prevent interaction of the arms with the core of the protomer. CONCLUSIONS: Hinge prolines can be considered as 'quaternary structure helpers'. The presence of a proline should be considered when searching for a determinant of oligomerization with arm exchange and could be used to engineer synthetic oligomers or to displace a monomers to oligomers equilibrium by mutation of this proline residue.

Crystallization and preliminary X-ray data of a phleomycin-binding protein from Streptoalloteichus hindustanus.

Large single crystals of a glycopeptide resistance protein encoded by the SH-ble gene from Streptoalloteichus hindustanus have been grown using vapor diffusion techniques with ammonium sulfate as the precipitant. The diffraction pattern extends to 2.0 A resolution. The crystals belong to space group P4(1)2(1)2 or P4(3)2(1)2 and have unit cell dimensions of a = 48.8 A and c = 112.5 A.

Site-directed mutagenesis of active site contact residues in a hydrolytic abzyme: evidence for an essential histidine involved in transition state stabilization.

Specific molecular interactions involved in catalysis by antibody 6D9 were investigated by site-directed mutagenesis. The catalytic antibody 6D9, which was generated against a transition state analog (III), hydrolyzes a non-bioactive chloramphenicol monoester derivative (I) to produce chloramphenicol (II). Construction of a three-dimensional molecular model of 6D9 and sequence comparison within a panel of related antibodies suggested candidates for catalytic residues, His (L27d), Tyr (L32), Tyr (H58) and Arg (H100b); these were targeted for the site-directed mutagenesis study. The Y-H58-F and R-H100b-A mutants possessed catalytic activities comparable to that of the wild-type, and the Y-H58-H and Y-L32-F mutant displayed an approximately fivefold decrease in k(cat)/Km. In the transition state analysis, the plots of logK(TSA) versus log(k(cat)/Km) for the mutants are linear, with a slope of approximately 1.0, indicating that the entire hapten-binding energy in the mutants is also utilized to bind the transition state and to accelerate the catalysis. In addition, a dramatic change in the catalytic activity was observed when the histidine residue (27d) in the CDR1 light chain was replaced with alanine. The H-L27d-A mutant had no detectable catalytic activity. This mutation led to a large, 40-fold reduction in transition state binding, with no change in substrate binding. Coupled with the previous kinetic studies and chemical modifications of the intact 6D9 antibody, this mutagenesis study has demonstrated that His L27d plays an essential role in stabilization of the transition state, the mechanism of catalysis by the 6D9 antibody.

A Monte Carlo based method to estimate radiation dose from multidetector CT (MDCT): cylindrical and anthropomorphic phantoms.

The purpose of this work was to extend the verification of Monte Carlo based methods for estimating radiation dose in computed tomography (CT) exams beyond a single CT scanner to a multidetector CT (MDCT) scanner, and from cylindrical CTDI phantom measurements to both cylindrical and physical anthropomorphic phantoms. Both cylindrical and physical anthropomorphic phantoms were scanned on an MDCT under the specified conditions. A pencil ionization chamber was used to record exposure for the cylindrical phantom, while MOSFET (metal oxide semiconductor field effect transistor) detectors were used to record exposure at the surface of the anthropomorphic phantom. Reference measurements were made in air at isocentre using the pencil ionization chamber under the specified conditions. Detailed Monte Carlo models were developed for the MDCT scanner to describe the x-ray source (spectra, bowtie filter, etc) and geometry factors (distance from focal spot to isocentre, source movement due to axial or helical scanning, etc). Models for the cylindrical (CTDI) phantoms were available from the previous work. For the anthropomorphic phantom, CT image data were used to create a detailed voxelized model of the phantom's geometry. Anthropomorphic phantom material compositions were provided by the manufacturer. A simulation of the physical scan was performed using the mathematical models of the scanner, phantom and specified scan parameters. Tallies were recorded at specific voxel locations corresponding to the MOSFET physical measurements. Simulations of air scans were performed to obtain normalization factors to convert results to absolute dose values. For the CTDI body (32 cm) phantom, measurements and simulation results agreed to within 3.5% across all conditions. For the anthropomorphic phantom, measured surface dose values from a contiguous axial scan showed significant variation and ranged from 8 mGy/100 mAs to 16 mGy/100 mAs. Results from helical scans of overlapping pitch (0.9375) and extended pitch (1.375) were also obtained. Comparisons between the MOSFET measurements and the absolute dose value derived from the Monte Carlo simulations demonstrate agreement in terms of absolute dose values as well as the spatially varying characteristics. This work demonstrates the ability to extend models from a single detector scanner using cylindrical phantoms to an MDCT scanner using both cylindrical and anthropomorphic phantoms. Future work will be extended to voxelized patient models of different sizes and to other MDCT scanners.

Characterization of the unlinked 16S rDNA and 23S-5S rRNA operon of Wolbachia pipientis, a prokaryotic parasite of insect gonads.

The rRNA-encoding genes (rDNAs) have been cloned and characterized from Wolbachia pipientis (Wp), the gonadial bacteria-like parasite of the mosquito Culex pipiens (Cp) and the moth Ephestia cautella (Ec). In Wp from both insect species the rDNAs are organized in a way which appears to be very unusual. The rRNAs are encoded by two unlinked transcription units, each present in a single copy per genome. One contains the 16S rDNA only, while the other is an operon encoding both the 23S and 5S rDNAs. Each transcription unit contains two putative upstream promoters, and downstream a Rho-independent terminator. The 16S rDNA, as well as the 23S-5S rRNA operon are not linked to any tRNA-encoding sequence and lack the antitermination boxes which are usually present immediately downstream from eubacterial promoters of rDNAs. Wp infecting Ec and Cp are highly similar taking as criteria the rDNAs and their flanking sequences. However, it clearly appears that each insect species harbours a different and specific Wp strain, or even subspecies. Phylogenetic relationships deduced from the complete sequences of their rDNAs undoubtedly confirm that Wp from Cp and Ec belong to the alpha-group of Proteobacteria, and are closely related to the Rickettsia.

Mammography film processor replenishment rate: bromide level monitoring.

The effects of the mammography film processing replenishment rate on contrast and speed are studied sensitometrically. Two experiments studied decreasing replenishment rates in the Kodak RP developer and quantified changes in the developer by measuring bromide ion concentrations. First, values of NaBr concentration from 1.7 to 8.4 g/L, achieved by reducing the replenishment rate, were tested with sensitometry strips. Second, the developer replenishment rate of a high volume dedicated mammography processor was reduced by one-third, to 20 cm3/1560 cm2, so that the NaBr concentration rose from 2.0 to 12.36. Sensitometric results for four film types and patient films were tested for changes from standard values as NaBr concentration was restored to 3.31 g/L. Fifty-five clinical images obtained at 7.3-9.3 NaBr g/L were compared to their matching previous films, with NaBr levels of 2-3 g/L, for contrast and visibility of the skin line. For the range of the NaBr ion from 1.7 to 7 g/L, no significant sensitometric differences were found. Above 7 g/L, different film types had different sensitometric results. From 7.3 to 9.3 NaBr g/L, 47.5% of the clinical films reviewed by four radiologists had less contrast compared to previous films. Dedicated mammography processors with high film volume (i.e., those that do not have excessive oxidation or foreign dye problems) can operate at lower replenishment rates than are currently employed. All common mammography film types are stable at these lower replenishment rates up to 7.0 NaBr g/L.

A Monte Carlo-based method to estimate radiation dose from spiral CT: from phantom testing to patient-specific models.

The purpose of this work is to develop and test a method to estimate the relative and absolute absorbed radiation dose from axial and spiral CT scans using a Monte Carlo approach. Initial testing was done in phantoms and preliminary results were obtained from a standard mathematical anthropomorphic model (MIRD V) and voxelized patient data. To accomplish this we have modified a general purpose Monte Carlo transport code (MCNP4B) to simulate the CT x-ray source and movement, and then to calculate absorbed radiation dose in desired objects. The movement of the source in either axial or spiral modes was modelled explicitly while the CT system components were modelled using published information about x-ray spectra as well as information provided by the manufacturer. Simulations were performed for single axial scans using the head and body computed tomography dose index (CTDI) polymethylmethacrylate phantoms at both central and peripheral positions for all available beam energies and slice thicknesses. For comparison, corresponding physical measurements of CTDI in phantom were made with an ion chamber. To obtain absolute dose values, simulations and measurements were performed in air at the scanner isocentre for each beam energy. To extend the verification, the CT scanner model was applied to the MIRD V model and compared with published results using similar technical factors. After verification of the model, the generalized source was simulated and applied to voxelized models of patient anatomy. The simulated and measured absolute dose data in phantom agreed to within 2% for the head phantom and within 4% for the body phantom at 120 and 140 kVp; this extends to 8% for the head and 9% for the body phantom across all available beam energies and positions. For the head phantom, the simulated and measured absolute dose data agree to within 2% across all slice thicknesses at 120 kVp. Our results in the MIRD phantom agree within 11% of all the different organ dose values published by the UK's ImPACT group for a scan using an equivalent scanner, kVp, collimation, pitch and mAs. The CT source model was shown to calculate both a relative and absolute radiation dose distribution throughout the entire volume in a patient-specific matrix geometry. Results of initial testing are promising and application to patient models was shown to be feasible.

Varying kVp as a means of reducing CT breast dose to pediatric patients.

We investigated the possibility of reducing radiation dose to the breast tissue of pediatric females by using multiple tube voltages within a single CT examination. The peak kilovoltage (kVp) was adjusted when the x-ray beam was directly exposing the representative breast tissue of a 5-year-old, 10-year-old, and an adult female anthropomorphic phantom; this strategy was called kVp splitting and was emulated by using a different kVp over the anterior and posterior tube angles. Dose savings from kVp splitting were calculated relative to using a fixed kVp over all tube angles and the results indicated savings in all three phantoms when using 80 kVp over the posterior tube angles regardless of the anterior kVp. Monte Carlo (MC) simulations with and without kVp splitting were performed to estimate absorbed breast dose in voxelized models constructed from the CT images of pediatric female patients; 80 kVp was used over the posterior tube angles. The MC simulations revealed breast dose savings of between 9.8% and 33% from using kVp splitting compared to simulations using a fixed kVp protocol with the anterior technique. Before this strategy could be implemented clinically, the development of suitable image reconstruction algorithms and the image quality of scans with kVp splitting would need further study.

A new family of sugar-inducible expression vectors for Escherichia coli.

A set of 11 expression vectors was constructed, each of them harbouring a cloning cassette under the control of the araB promoter. Some of these vectors enable expression of foreign proteins in the cytoplasm, while others include a synthetic sequence coding for a very efficient secretion signal sequence. Other features are an f1 origin of replication (in plus or minus orientation) and a promoter(up) mutation that enhances the already very high level of expression from these vectors. With such a versatile vector family, cloning, sequencing and site-directed mutagenesis can be performed on the same vector, and the level of expression can be defined according to the specific constraints of a given protein.

Crystal structure and site-directed mutagenesis of a bleomycin resistance protein and their significance for drug sequestering.

The antibiotic bleomycin, a strong DNA cutting agent, is naturally produced by actinomycetes which have developed a resistance mechanism against such a lethal compound. The crystal structure, at 2.3 A resolution, of a bleomycin resistance protein of 14 kDa reveals a structure in two halves with the same alpha/beta fold despite no sequence similarity. The crystal packing shows compact dimers with a hydrophobic interface and involved in mutual chain exchange. Two independent solution studies (analytical centrifugation and light scattering) showed that this dimeric form is not a packing artefact but is indeed the functional one. Furthermore, light scattering also showed that one dimer binds two antibiotic molecules as expected. A crevice located at the dimer interface, as well as the results of a site-directed mutagenesis study, led to a model wherein two bleomycin molecules are completely sequestered by one dimer. This provides a novel insight into antibiotic resistance due to drug sequestering, and probably also into drug transport and excretion.

CM-gag, a transposable-like element reiterated in the genome of Culex pipiens mosquitoes, contains only a gag gene.

CM-gag elements constitute an homogeneous family of sequences that are reiterated in the genome of Culex pipiens strains from different continents. Apparently complete 1.75 kb CM-gag copies are flanked by target-site duplications and have a polyadenylation signal near their 3' end. They potentially contain a unique gene encoding a putative protein that displays homologies with nucleic acid binding proteins and the gag polypeptide of retroviruses and retrotransposons, but that does not encode a reverse transcriptase. CM-gag elements are similar in their genetic organization to the telomeric transposable sequences Het-A from Drosophila melanogaster, but Southern-hybridization patterns indicate that the former are more probably dispersed in various areas of the mosquito genome. The homogeneity of CM-gag copies that are distributed worldwide suggests that they have most probably been amplified recently. Furthermore, selective constraints against amino acid changes have been acting on these sequences, suggesting that they need to encode the gag-like protein to be incorporated into the chromosomes.

Exposure rates in high-level-control fluoroscopy for image enhancement.

High-level fluoroscopic boost options that exceed conventional exposure limits are available as a means of reducing quantum mottle during angiography. Federal law does not specify exposure limits for such high-level controls but requires specific means of activation to safeguard against inadvertent use. The American Association of Physicists in Medicine recently recommended that high-level exposure rates not exceed 2.58 mC/kg/min (10R/min). At six institution surveyed, maximum exposure rates ranged from 5.42 to 24 mC/kg/min (21-93 R/min). Activation of high-level capability varied from a simple foot switch to a keyed interlock requiring a second operator to engage. There appears to be no industry coherence in high-level control exposure limits as yet, although the Center for Devices and Radiological Health recently initiated an investigatory program.