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Biomedical subjects

David L Wilson

Publications and source records attributed to David L Wilson.

36 records · Page 2Linked to original sources

Radio-frequency-induced thermal lesions: subacute magnetic resonance appearance and histological correlation.

PURPOSE: To investigate the relationship between subacute magnetic resonance (MR) images of radio-frequency (RF) ablation lesions and tissue viability as determined from histological tissue samples. MATERIALS AND METHODS: We generated lesions (N = 5) in a rabbit thigh model. Four days later, we obtained in vivo T(2)- and contrast-enhanced (CE) T(1)-weighted images and ex vivo histological samples approximately perpendicular to the electrode path. Using three-dimensional registration and warping, we spatially compared manually segmented boundaries apparent on MR images to boundaries separating distinct histological zones determined from hematoxylin and eosin (H&E) and Masson trichrome (MT) stains, as well as birefringence studies. RESULTS: Lesions have a characteristic MR appearance: an outer hyperintense margin (M2) separating background tissue (M3) from an inner core (M1), in both T(2) and CE T(1) images. Histologically, there are two zones of damage: an outer zone of likely nonviable cells (H2) separating background tissue (H3) from an inner core of coagulated nonviable cells (H1). We measured distances between automatically computed correspondence points along histological and MR boundaries. For T(2) and CE T(1) images, respectively, M1 vs. H1 distances were 0.72 +/- 0.99 mm (mean +/- SD) and 0.10 +/- 0.95 mm, while outer M2 vs. H2 boundary distances were 0.26 +/- 1.16 mm and 0.05 +/- 1.08 mm. The discrepancy between histological and MR boundaries was larger than the variability in segmenting MR images, but probably within registration error. There were no significant differences between T(2) and CE T(1) boundaries. CONCLUSION: Lesion boundaries apparent in both T(2)- and CE T(1)-weighted MR scans, performed several days postablation, similarly predict the histological response. That is, the lesion core (M1) corresponds to nonviable coagulated cells (H1), while the hyperintense margin (M2) corresponds to likely nonviable cells undergoing necrotic changes (H2).

Animals↗

Noninvasive monitoring of local drug release using X-ray computed tomography: optimization and in vitro/in vivo validation.

In vivo release profiles of drug-loaded biodegradable implants were noninvasively monitored and characterized using X-ray computed tomography (CT). The imaging method was adapted and optimized to quantitatively examine the release of an active agent from a model cylindrical PLGA device (the millirod) into rabbit livers over 48 h. Iohexol, a CT contrast agent, served as a model drug; optimization of CT acquisition parameters yielded a sensitivity of 0.21 mg/mL (or 95 microg iodine/mL) for this agent. In vitro validation of the method was carried out by tracking release of iohexol in gelatin gel phantoms. In vivo release in rabbit livers was characterized through quantitative analysis of CT images and compared with UV-Vis analysis of the explanted devices at three implantation times. After correction for respiratory motion, CT analysis correlates well with the extracted iohexol data at all time points. The percent error between the actual and experimental image data was below 10%. This study demonstrates the potential of using computed tomography to noninvasively quantify the rate of agent release from controlled delivery devices in vivo.

Animals↗

Automatic registration of MR and SPECT images for treatment planning in prostate cancer.

RATIONALE AND OBJECTIVES: To aid in surgical and radiation therapy planning for prostate adenocarcinoma, a general-purpose automatic registration method that is based on mutual information was used to align magnetic resonance (MR) images and single photon emission computed tomographic (SPECT) images of the pelvis and prostate. MATERIALS AND METHODS: The authors assessed the effects of various factors on alignment between pairs of MR and SPECT images, including the use of particular pulse sequences in MR imaging, image voxel intensity scaling, the use of different regions on the MR-SPECT histogram, spatial masking of nonoverlapping visual data between images, and multiresolution optimization. A mutual information algorithm was used as the cost function for automatic registration. Automatic registration was deemed acceptable when it resulted in a transformation with less than 2 voxel units (6 mm) difference in translation and less than 2 degree difference in rotation from that obtained with manual registration performed independently by nuclear medicine radiologists. RESULTS: Paired sets of MR and SPECT image volumes from four of five patients were successfully registered. For successful registration, MR images must be optimal and registration must be performed at full spatial resolution and at the full intensity range. Masking, cropping, and the normalization of mutual information, used to register partially overlapping MR-SPECT volumes, were not successful. Multiresolution optimization had little effect on the accuracy and speed of the registration. CONCLUSION: Automatic registration between MR and SPECT images of the pelvis can be achieved when data acquisition and image processing are performed properly. It should prove useful for prostate cancer diagnosis, staging, and treatment planning.

Adenocarcinoma↗

Variation of the natural transformation frequency of Campylobacter jejuni in liquid shake culture.

Natural transformation, a mechanism that generates genetic diversity in Campylobacter jejuni, was studied in a novel liquid shake culturing system that allowed an approximately 10 000-fold increase in cell density. C. jejuni transformation frequency was analysed in this system under 10 %, 5.0 % and 0.7 % CO(2) atmospheres. At 5.0 % and 10 % CO(2) concentrations, when purified isogenic chromosomal DNA was used to assess competence, transformation frequency ranged from 10(-3) to 10(-4) at low cell concentrations and declined as cell density increased. Transformation frequency under a 0.7 % CO(2) atmosphere was more stable, maintaining 10(-3) levels at high cell densities, and was 10- to 100-fold higher than that under a 10 % CO(2) atmosphere. Three of four C. jejuni strains tested under a 5.0 % CO(2) atmosphere were naturally competent for isogenic DNA; competent strains demonstrated a lack of barriers to intraspecies genetic exchange by taking up and incorporating chromosomal DNA from multiple C. jejuni donors. C. jejuni showed a preference for its own DNA at the species level, and co-cultivation demonstrated that DNA transfer via natural transformation occurred between isogenic populations during short periods of exposure in liquid medium when cell density and presumably DNA concentrations were low. Transformation frequency during co-cultivation of isogenic populations was also influenced by CO(2) concentration. Under a 0.7 % CO(2) atmosphere, co-cultivation transformation frequency increased approximately 500-fold in a linear fashion with regard to cell density, and was 1000- to 10 000-fold higher during late-exponential-phase growth when compared to cultures grown under a 10 % CO(2) atmosphere.

Bacteriological Techniques↗

Slice-to-volume registration and its potential application to interventional MRI-guided radio-frequency thermal ablation of prostate cancer.

In this study, we registered live-time interventional magnetic resonance imaging (iMRI) slices with a previously obtained high-resolution MRI volume that in turn can be registered with a variety of functional images, e.g., PET, SPECT, for tumor targeting. We created and evaluated a slice-to-volume (SV) registration algorithm with special features for its potential use in iMRI-guided radio-frequency (RF) thermal ablation of prostate cancer. The algorithm features included a multiresolution approach, two similarity measures, and automatic restarting to avoid local minima. Imaging experiments were performed on volunteers using a conventional 1.5-T MR scanner and a clinical 0.2-T C-arm iMRI system under realistic conditions. Both high-resolution MR volumes and actual iMRI image slices were acquired from the same volunteers. Actual and simulated iMRI images were used to test the dependence of SV registration on image noise, receive coil inhomogeneity, and RF needle artifacts. To quantitatively assess registration, we calculated the mean voxel displacement over a volume of interest between SV registration and volume-to-volume registration, which was previously shown to be quite accurate. More than 800 registration experiments were performed. For transverse image slices covering the prostate, the SV registration algorithm was 100% successful with an error of <2 mm, and the average and standard deviation was only 0.4 mm +/- 0.2 mm. Visualizations such as combined sector display and contour overlay showed excellent registration of the prostate and other organs throughout the pelvis. Error was greater when an image slice was obtained at other orientations and positions, mostly because of inconsistent image content such as that from variable rectal and bladder filling. These preliminary experiments indicate that MR SV registration is sufficiently accurate to aid image-guided therapy.

Algorithms↗

Volume registration using needle paths and point landmarks for evaluation of interventional MRI treatments.

We created a method for three-dimensional (3-D) registration of medical images (e.g., magnetic resonance imaging (MRI) or computed tomography) to images of physical tissue sections or to other medical images and evaluated its accuracy. Our method proved valuable for evaluation of animal model experiments on interventional-MRI guided thermal ablation and on a new localized drug delivery system. The method computes an optimum set of rigid body registration parameters by minimization of the Euclidean distances between automatically chosen correspondence points, along manually selected fiducial needle paths, and optional point landmarks, using the iterative closest point algorithm. For numerically simulated experiments, using two needle paths over a range of needle orientations, mean voxel displacement errors depended mostly on needle localization error when the angle between needles was at least 20 degrees. For parameters typical of our in vivo experiments, the mean voxel displacement error was < 0.35 mm. In addition, we determined that the distance objective function was a useful diagnostic for predicting registration quality. To evaluate the registration quality of physical specimens, we computed the misregistration for a needle not considered during the optimization procedure. We registered an ex vivo sheep brain MR volume with another MR volume and tissue section photographs, using various combinations of needle and point landmarks. Mean registration error was always < or = 0.54 mm for MR-to-MR registrations and < or = 0.52 mm for MR to tissue section registrations. We also applied the method to correlate MR volumes of radio-frequency induced thermal ablation lesions with actual tissue destruction. In this case, in vivo rabbit thigh volumes were registered to photographs of ex vivo tissue sections using two needle paths. Mean registration errors were between 0.7 and 1.36 mm over all rabbits, the largest error less than two MR voxel widths. We conclude that our method provides sufficient spatial correspondence to facilitate comparison of 3-D image data with data from gross pathology tissue sections and histology.

Anatomy, Cross-Sectional↗

Determination of ciprofloxacin and nalidixic acid resistance in Campylobacter jejuni with a fluorogenic polymerase chain reaction assay.

A fluorogenic polymerase chain reaction assay for the gyrA gene was used to determine the frequency of a Thr-86 mutation in Campylobacter jejuni isolates from food animals and humans in northern Thailand and to investigate the correlation between this mutation and bacterial resistance to fluoroquinolones. Eighty-four isolates of C. jejuni were used: 65 from healthy chickens on farms, 16 from chickens at the slaughterhouse, 1 from chicken meat at the market, and 1 from a healthy farm worker. The microbroth dilution technique was used for in vitro susceptibility testing. MIC breakpoints established by the National Antimicrobial Resistance Monitoring System were used to categorize the resistance of C. jejuni to ciprofloxacin and nalidixic acid. Sixty of the 84 C. jejuni isolates tested carried the Thr-86 mutation in the gyrA gene. All isolates with ciprofloxacin MICs of > or = 2 mg/liter carried the mutation, and no isolates with nalidixic acid MICs of < or = 16 mg/liter carried the Thr-86-to-Ile mutation. There was a very strong association between ciprofloxacin resistance and the presence of the mutation (kappa = 0.971, P < 0.01). The association between the presence of the Thr-86-to-Ile mutation and nalidixic acid resistance was weaker (kappa 0.859: P < or = 0.01).

Abattoirs↗

Noninvasive monitoring of local drug release in a rabbit radiofrequency (RF) ablation model using X-ray computed tomography.

In this study, X-ray computed tomography (CT) was utilized as a noninvasive method to directly examine local drug release kinetics in livers before and following radiofrequency thermal ablation. Iohexol, a CT contrast agent, was used as a drug-mimicking molecule. Release of iohexol in healthy and ablated rabbit livers over 48 h was quantified and correlated with the release profiles from phosphate-buffered saline (PBS) in vitro. The results show that iohexol release in ablated livers is significantly slower than both release in normal livers and in vitro. The time at which 50% of the drug was released (t(1/2)) into ablated liver (20.6+/-5.9 h) was 1.7 times longer than in normal liver (12.1+/-5.4 h) and approximately two times longer than that in PBS (10.1+/-1.2 h). The slower release in ablated livers is a result of severe tissue damage inflicted by thermal ablation, as supported by histological examination. This data suggests that a noninvasive imaging method provides a superior measurement over in vitro release studies in accurately quantifying the local release kinetics of an agent in an altered physiological system in vivo. Because the development of a successful local drug therapy is dependent on the understanding of the agent release kinetics at the implantation site, the noninvasive data may be indispensable in effectively predicting the implant behavior in a physiological system.

Animals↗

Automatic MR volume registration and its evaluation for the pelvis and prostate.

A three-dimensional (3D) mutual information registration method was created and used to register MRI volumes of the pelvis and prostate. It had special features to improve robustness. First, it used a multi-resolution approach and performed registration from low to high resolution. Second, it used two similarity measures, correlation coefficient at lower resolutions and mutual information at full resolution, because of their particular advantages. Third, we created a method to avoid local minima by restarting the registration with randomly perturbed parameters. The criterion for restarting was a correlation coefficient below an empirically determined threshold. Experiments determined the accuracy of registration under conditions found in potential applications in prostate cancer diagnosis, staging, treatment and interventional MRI (iMRI) guided therapies. Images were acquired in the diagnostic (supine) and treatment position (supine with legs raised). Images were also acquired as a function of bladder filling and the time interval between imaging sessions. Overall studies on three patients and three healthy volunteers, when both volumes in a pair were obtained in the diagnostic position under comparable conditions, bony landmarks and prostate 3D centroids were aligned within 1.6 +/- 0.2 mm and 1.4 +/- 0.2 mm, respectively, values only slightly larger than a voxel. Analysis suggests that actual errors are smaller because of the uncertainty in landmark localization and prostate segmentation. Between the diagnostic and treatment positions, bony landmarks continued to register well, but prostate centroids moved towards the posterior 2.8-3.4 mm. Manual cropping to remove voxels in the legs was necessary to register these images. In conclusion, automatic, rigid body registration is probably sufficiently accurate for many applications in prostate cancer. For potential iMRI-guided treatments, the small prostate displacement between the diagnostic and treatment positions can probably be avoided by acquiring volumes in similar positions and by reducing bladder and rectal volumes.

Algorithms↗

Automatic 3D registration for interventional MRI-guided treatment of prostate cancer.

The goal of this research is to register real-time interventional magnetic resonance imaging (iMRI) slice images with a previously obtained high-resolution MRI image volume, which in turn can be registered with functional images such as those from SPECT. The immediate application is in iMRI-guided treatment of prostate cancer, where additional images are desired to improve tumor targeting. In this article, simulation experiments are performed to demonstrate the feasibility of slice-to-volume registration for this application. We acquired 3D volume images from a 1.5-T MRI system and simulated low-field iMRI image slices by creating thick slices and adding noise. We created a slice-to-volume mutual information registration algorithm with special features to improve robustness. Features included a multiresolution approach, two similarity measures, and automatic restarting to avoid local minima. To assess the quality of registration, we calculated 3D displacements on a voxel-by-voxel basis over a volume of interest between slice-to-volume registration and volume-to-volume registration, which was previously shown to be quite accurate. More than 800 registration experiments were performed on MR images of three volunteers. The slice-to-volume registration algorithm was very robust and accurate for transverse slice images covering the prostate, with a registration error of only 0.4 +/- 0.2 mm. Error was greater at other slice orientations and positions. The automatic slice-to-volume mutual information registration algorithm is robust and probably sufficiently accurate to aid in iMRI-guided treatment of prostate cancer.

Algorithms↗

Three-dimensional model of lesion geometry for evaluation of MR-guided thermal ablation therapy.

RATIONALE AND OBJECTIVES: High-radiofrequency energy is used clinically to ablate pathologic tissue with interventional magnetic resonance (MR) imaging. For many tissues, resulting lesions have a characteristic appearance on contrast-enhanced T1- and T2-weighted MR images, with two boundaries enclosing an inner hypointense region and an outer hyperintense margin. Geometric modeling of three-dimensional thermal lesions in animal experiments and patient treatments would improve analyses and visualization. MATERIALS AND METHODS: The authors created a model with two quadric surfaces and 12 parameters to describe both lesion surfaces. Parameters were estimated with iterative optimization to minimize the sum of the squared shortest distances from segmented points to the model surface. The authors validated the estimation process with digital lesion phantoms that simulated varying levels of segmentation error and missing surface information. They also applied their method to in vivo images of lesions in a rabbit model. RESULTS: For simulated phantom lesions, the lesion geometry was accurate despite manual segmentation error and incomplete surface data. Even when 50% of the surface was missing, the median error was less than 0.5 mm. For all in vivo lesions, the median distance from the model surface to data was no more than 0.58 mm for both inner and outer surfaces, less than a voxel width (0.7 mm). The interquartile range was 0.89 mm or less for all data. CONCLUSION: The authors' model provides a good approximation of actual lesion geometry and is highly resistant to missing segmentation information. It should prove useful for three-dimensional lesion visualization, volume estimation, automated segmentation, and volume registration.

Abdominal Muscles↗

X-ray computed tomography methods for in vivo evaluation of local drug release systems.

Recent advances in drug delivery techniques have necessitated the development of tools for in vivo monitoring of drug distributions. Gamma emission imaging and magnetic resonance imaging suffer from problems of resolution and sensitivity, respectively. We propose that the combination of X-ray CT imaging and image analysis techniques provides an excellent method for the evaluation of the transport of platinum-containing drugs from a localized, controlled release source. We correlated local carboplatin concentration with CT intensity, producing a linear relationship with a sensitivity of 62.6 microg/mL per Hounsfield unit. As an example application, we evaluated the differences in drug transport properties between normal and ablated rabbit liver from implanted polymer millirods. The use of three-dimensional visualization provided a method of evaluating the placement of the drug delivery device in relation to the surrounding anatomy, and registration and reformatting allowed the accurate comparison of the sequence of temporal CT volumes acquired over a period of 24 h. Taking averages over radial lines extending away from the center of the implanted millirods and integrating over clinically appropriate regions, yielded information about drug release from the millirod and transport in biological tissues. Comparing implants in normal and ablated tissues, we found that ablation prior to millirod implantation greatly decreased the loss of drug from the immediate area, resulting in a higher average dose to the surrounding tissue. This work shows that X-ray CT imaging is a useful technique for the in vivo evaluation of the pharmacokinetics of platinated agents.

Animals↗

Image quality evaluation of flat panel and image intensifier digital magnification in x-ray fluoroscopy.

Interventional devices used in radiology often have dimensions on the order of a pixel, and radiologists resort to image magnification to better visualize such small devices. Traditional image intensifier (II) systems use analog magnification with x-ray exposure inversely proportional to the area of field of view (FOV) so as to maintain light output for the camera. Analog magnification is impossible with flat panel (FP) detectors, and images must be magnified using digital interpolation that does not reduce the pixel partial area effect for small devices. We quantitatively investigated image quality of digital and analog magnification using a clinically relevant task of detecting a partially deployed stent in x-ray fluoroscopy image sequences that were created using realistic detector models. Using the standard exposure strategy for II analog magnification, exposure was increased from a nominal 43.65 nGy (5.0 microR) per frame at 23 cm FOV to 79.9 nGy (9.15 microR) per frame and 117.81 nGy (13.49 microR) per frame at 17 cm and 14 cm FOV, respectively. Contrast sensitivity improved significantly (p<0.1) by 43.5+/-6.5% and 64.1+/-7.3% with the 17 cm and 14 cm FOV, respectively. Exposure for digitally magnified images was varied in an adaptive forced choice experiment so as to match performance with II analog magnification. For digital magnification, bilinear interpolation was used to give magnified stents sizes equivalent to those in the analog magnified images. For equivalent image quality, FP required 34.87+/-2.59, 80.16+/-5.37, and 84.08+/-5.59 nGy per frame at normal, and the two magnification modes, respectively. Hence, FP with digital magnification gives significant (p<0.1) dose savings of 20+/-6% and 27+/-5% at the normal and highest magnification modes, respectively. Digitally magnified II images required exposures of 110.85+/-8.07 and 103.34+/-5.90 nGy per frame for the two magnifications levels, respectively, giving no significant (p>0.1) dose savings. A spatiotemporal human observer model based on signal detection theory successfully predicted the human data and was used to predict other conditions associated with image magnification. Model predictions quantitatively showed that magnification is most useful when signal size is relatively small and that FP digital magnification can improve image quality for the stent deployment task without increasing exposure. In conclusion, the results show that FP digital magnification can be useful and dose efficient as compared to analog magnification.

Humans↗

Chromosomal location plays a role in regulation of aflatoxin gene expression in Aspergillus parasiticus.

The nor-1 gene in the filamentous fungus Aspergillus parasiticus encodes a ketoreductase involved in aflatoxin biosynthesis. To study environmental influences on nor-1 expression, we generated plasmid pAPGUSNNB containing a nor-1 promoter-beta-glucuronidase (GUS) (encoded by uidA) reporter fusion with niaD (encodes nitrate reductase) as a selectable marker. niaD transformants of A. parasiticus strain NR-1 (niaD) carried pAPGUSNNB integrated predominantly at the nor-1 or niaD locus. Expression of the native nor-1 and nor-1::GUS reporter was compared in transformants grown under aflatoxin-inducing conditions by Northern and Western analyses and by qualitative and quantitative GUS activity assays. The timing and level of nor-1 promoter function with pAPGUSNNB integrated at nor-1 was similar to that observed for the native nor-1 gene. In contrast, nor-1 promoter activity in pAPGUSNNB and a second nor-1::GUS reporter construct, pBNG3.0, was not detectable when integration occurred at niaD. Because niaD-dependent regulation could account for the absence of expression at niaD, a third chromosomal location was analyzed using pAPGUSNP, which contained nor-1::GUS plus pyrG (encodes OMP decarboxylase) as a selectable marker. GUS expression was detectable only when pAPGUSNP integrated at nor-1 and was not detectable at pyrG, even under growth conditions that required pyrG expression. nor-1::GUS is regulated similarly to the native nor-1 gene when it is integrated at its homologous site within the aflatoxin gene cluster but is not expressed at native nor-1 levels at two locations outside of the aflatoxin gene cluster. We conclude that the GUS reporter system can be used effectively to measure nor-1 promoter activity and that nor-1 is subject to position-dependent regulation in the A. parasiticus chromosome.

Aflatoxins↗

Quantitative assessment of image quality enhancement due to unsharp-mask processing in x-ray fluoroscopy.

Spatial unsharp-mask processing and its variants are commonly used in x-ray radiography to enhance image contrast. We investigated the effect of three unsharp-masking filter kernels of different sizes on the detection of an advanced guidewire tip in simulated x-ray fluoroscopy image sequences. To isolate the effect of visual temporal processing, we repeated the experiments on single images. Filter gains were selected so that all three kernels increased the contrast of a 0.018-in. (0.457-mm) guidewire by a factor of 2 but had different effects on image noise and signal profiles. There was no statistically significant effect of unsharp masking on human-observer performance in single images. However, all three kernels significantly improved average performance in image sequences, and the guidewire contrast required for detection was reduced by 32%-40%. A prewhitening channelized observer model predicted the disparity between sequences and single images and fitted measurements at different kernel sizes well. A nonprewhitening observer model did not. We conclude that unsharp masking is a simple and effective method of improving guidewire visualization in fluoroscopically guided interventional procedures and that quantitative image quality studies are essential for evaluation of image-processing techniques in sequences such as x-ray fluoroscopy.

Adult↗

Identification of Campylobacter jejuni isolates from cloacal and carcass swabs of chickens in Thailand by a 5' nuclease fluorogenic polymerase chain reaction assay.

A rapid 5' nuclease fluorogenic polymerase chain reaction (PCR) assay for identifying Campylobacter jejuni was applied to Campylobacter isolates from chicken cloacal and carcass swabs collected from three chicken farms and a slaughterhouse in Thailand. The primers and the probe were based on the sequence of the gyrA gene in C jejuni. C. jejuni isolates were identified by fluorogenic PCR assay of bacterial cells directly from Campylobacter-selective agar medium. This assay allowed the identification of C. jejuni within 1 day after colonies appeared on selective media. The fluorogenic PCR assay yielded results comparable to those of the conventional test kit (kappa = 0.76) but required less time. When the two methods disagreed with regard to species identification, results were confirmed by PCR restriction fragment length polymorphism of 23S rRNA genes. In these instances, the fluorogenic PCR assay correctly identified more isolates of C. jejuni than did the conventional test kit (six of seven isolates were unidentifiable by the conventional test kit). The fluorogenic PCR assay is a rapid and specific method that outperforms the conventional test kit in the identification of C. jejuni from environmental samples.

Animals↗

A comparative study of warping and rigid body registration for the prostate and pelvic MR volumes.

A three-dimensional warping registration algorithm was created and compared to rigid body registration of magnetic resonance (MR) pelvic volumes including the prostate. The rigid body registration method combines the advantages of mutual information (MI) and correlation coefficient at different resolutions. Warping registration is based upon independent optimization of many interactively placed control points (CP's) using MI and a thin plate spline transformation. More than 100 registration experiments with 17 MR volume pairs determined the quality of registration under conditions simulating potential interventional MRI-guided treatments of prostate cancer. For image pairs that stress rigid body registration (e.g. supine, the diagnostic position, and legs raised, the treatment position), both visual and numerical evaluation methods showed that warping consistently worked better than rigid body. Experiments showed that approximately 180 strategically placed CP's were sufficiently expressive to capture important features of the deformation.

Algorithms↗

Sustained in vivo gene delivery from agarose hydrogel prolongs nonviral gene expression in skin.

Prolonging gene expression in skin using safe, nonviral gene delivery techniques could impact skin regeneration and wound healing, decrease infection, and potentially improve the success of tissue-engineered skin. To this end, an injectable, agarose-based delivery system was tested and shown to prolong nonviral gene expression in the skin. DNA was compacted with polylysine to improve DNA stability in the presence of nucleases. Up to 25 microg of compacted luciferase plasmid with or without agarose hydrogel was injected intradermally in rodents. Bioluminescence imaging was used for longitudinal, noninvasive monitoring of gene expression in vivo for 35 days. Injections of DNA in solution produced gene expression for only 5-7 days, whereas the sustained release of compacted DNA from the agarose system prolonged expression, with more than 500 pg (20% of day 1 levels) of luciferase per site for at least 35 days. Southern blotting confirmed that the agarose system extended DNA retention, with significant plasmid present through day 7, as compared with DNA in solution, which had detectable DNA only on day 1. Histology revealed that agarose invoked a wound-healing response through day 14. Tissue-engineering and wound-healing applications may benefit from the agarose gene delivery system.

Absorbable Implants↗