Reliability of Computer-Assisted Breast Density Estimation: Comparison of Interactive Thresholding, Semiautomated, and Fully Automated Methods.

OBJECTIVE: The purpose of this study was to investigate the reliability of computer-assisted methods of estimating breast density. MATERIALS AND METHODS: Craniocaudal mammograms of 100 healthy subjects were collected from a screening mammography database. Three expert readers independently assessed mammographic breast density twice in a 1-month period using interactive thresholding and semiautomated methods. In addition, fully automated breast density estimation software was used to generate objective breast density estimates. The reliability of the computer-assisted breast density estimation was assessed in terms of concordance correlation coefficients, limits of agreement, systematic difference, and reader variability. RESULTS: Statistically significant systematic bias (paired t test, p < 0.01) and variability (4.75-10.91) were found within and between readers for both the interactive thresholding and the semiautomated methods. Using the semiautomated method significantly reduced the within-reader bias of one reader (p < 0.02) and the between-reader variability of all three readers (p < 0.05). The breast density estimates obtained with the fully automated method had excellent agreement with those of the reference standard (concordance correlation coefficient, 0.93) without a significant systematic difference. CONCLUSION: Reader-dependent variability and systematic bias exist in breast density estimates obtained with the interactive thresholding method, but they may be reduced in part by use of the semiautomated method. Assessing reader performance may be necessary for more reliable breast density estimation, especially for surveillance of breast density over time. The fully automated method has the potential to provide reliable breast density estimates nearly free from reader-dependent systematic bias and reader variability.

Efficacy of Selective Brain Cooling Using a Nasopharyngeal Method in Piglets.

BACKGROUND: Mild hypothermia is an effective neuroprotective strategy for a variety of acute brain injuries. Cooling the nasopharynx may offer the capability to cool the brain selectively due to anatomic proximity of the internal carotid artery to the cavernous sinus. This study investigated the feasibility and efficiency of nasopharyngeal brain cooling by continuously blowing room temperature or cold air at different flow rates into the nostrils of normal newborn piglets. METHODS: Experiments were conducted on thirty piglets (n = 30, weight = 2.7 ± 1.5 kg). Piglets were anesthetized with 1­2% isoflurane and were randomized to receive one of four different nasopharyngeal cooling treatments: I. Room temperature at a flow rate of 3­4 L min(−1) (n = 6); II. −1 ± 2 °C at a flow rate of 3­4 L min(−1) (n = 6); III. Room temperature at a flow rate of 14­15 L min(−1) (n = 6); IV. −8 ± 2 °C at a flow rate of 14­15 L min(−1) (n = 6). To control for the normal thermal regulatory response of piglets without nasopharyngeal cooling, a control group of piglets (n = 6) had their brain temperature monitored without nasopharyngeal cooling. The duration of treatment was 60 min, with additional 30 min of observation. RESULTS: In group I, median cooling rate was 1.7 ± 0.9 °C/h by setting the flow rate of room temperature air to 3­4 L min(−1). Results of comparing different temperatures and flow rates in the nasopharyngeal cooling approach reveal that the brain temperature could be reduced rapidly at a rate of 5.5 ± 1.1 °C/h by blowing −8 ± 2 °C air at a flow rate of 14­15 L min(−1). CONCLUSIONS: Nasopharyngeal cooling via cooled insufflated air can lower the brain temperature, with higher flows and lower temperatures of insufflated air being more effective.

Object detection and tracking using a high-performance artificial intelligence-based 3D depth camera: towards early detection of African swine fever.

BACKGROUND: Inspection of livestock farms using surveillance cameras is emerging as a means of early detection of transboundary animal disease such as African swine fever (ASF). Object tracking, a developing technology derived from object detection aims to the consistent identification of individual objects in farms. OBJECTIVES: This study was conducted as a preliminary investigation for practical application to livestock farms. With the use of a high-performance artificial intelligence (AI)-based 3D depth camera, the aim is to establish a pathway for utilizing AI models to perform advanced object tracking. METHODS: Multiple crossovers by two humans will be simulated to investigate the potential of object tracking. Inspection of consistent identification will be the evidence of object tracking after crossing over. Two AI models, a fast model and an accurate model, were tested and compared with regard to their object tracking performance in 3D. Finally, the recording of pig pen was also processed with aforementioned AI model to test the possibility of 3D object detection. RESULTS: Both AI successfully processed and provided a 3D bounding box, identification number, and distance away from camera for each individual human. The accurate detection model had better evidence than the fast detection model on 3D object tracking and showed the potential application onto pigs as a livestock. CONCLUSIONS: Preparing a custom dataset to train AI models in an appropriate farm is required for proper 3D object detection to operate object tracking for pigs at an ideal level. This will allow the farm to smoothly transit traditional methods to ASF-preventing precision livestock farming.

Imaging of gene expression in live pancreatic islet cell lines using dual-isotope SPECT.

UNLABELLED: We are combining nuclear medicine with molecular biology to establish a sensitive, quantitative, and tomographic method with which to detect gene expression in pancreatic islet cells in vivo. Dual-isotope SPECT can be used to image multiple molecular events simultaneously, and coregistration of SPECT and CT images enables visualization of reporter gene expression in the correct anatomic context. We have engineered pancreatic islet cell lines for imaging with SPECT/CT after transplantation under the kidney capsule. METHODS: INS-1 832/13 and alphaTC1-6 cells were stably transfected with a herpes simplex virus type 1-thymidine kinase-green fluorescent protein (HSV1-thymidine kinase-GFP) fusion construct (tkgfp). After clonal selection, radiolabel uptake was determined by incubation with 5-(131)I-iodo-1-(2-deoxy-2-fluoro-beta-d-arabinofuranosyl)uracil ((131)I-FIAU) (alphaTC1-6 cells) or (123)I-FIAU (INS-1 832/13 cells). For the first set of in vivo experiments, SPECT was conducted after alphaTC1-6/tkgfp cells had been labeled with either (131)I-FIAU or (111)In-tropolone and transplanted under the left kidney capsule of CD1 mice. Reconstructed SPECT images were coregistered to CT. In a second study using simultaneous acquisition dual-isotope SPECT, INS-1 832/13 clone 9 cells were labeled with (111)In-tropolone before transplantation. Mice were then systemically administered (123)I-FIAU and data for both (131)I and (111)In were acquired simultaneously. RESULTS: alphaTC1-6/tkgfp cells showed a 15-fold greater uptake of (131)I-FIAU, and INS-1/tkgfp cells showed a 12-fold greater uptake of (123)I-FIAU, compared with that of wild-type cells. After transplantation under the kidney capsule, both reporter gene expression and location of cells could be visualized in vivo with dual-isotope SPECT. Immunohistochemistry confirmed the presence of glucagon- and insulin-positive cells at the site of transplantation. CONCLUSION: Dual-isotope SPECT is a promising method to detect gene expression in and location of transplanted pancreatic cells in vivo.

Isolating human islets of Langerhans causes loss of decay accelerating factor (CD55) on beta-cells.

It has previously been reported that human decay accelerating factor (DAF; CD55) is not expressed on cells isolated from human islets. We have investigated if this absence is caused by the islet isolation procedure and/or the single cell isolation technique. We focused on loss of DAF expression on beta-cells within the intact islet and on isolated individual beta-cells. We established that DAF was expressed in islets and on beta-cells prior to isolation by in situ analysis in the intact pancreas. In situ immunohistochemistry (IHC) was used to examine DAF expression on human pancreatic islets and isolated islets. A reverse transcriptase-polymerase chain reaction (RT-PCR) specific for human DAF mRNA was developed to measure mRNA levels in situ in islets within the intact pancreas, isolated islets, and purified beta-cells. beta-Cells were purified by fluorescence-activated cell sorting. DAF protein expression on these purified cells was measured using flow cytometry. Expression of DAF protein was present on the islets, including beta-cells within the human pancreas; however, comparative data from IHC and flow cytometry revealed the absence of DAF protein on beta-cells in both isolated islets and single cell preparations. Furthermore, compared to mRNA levels detected by in situ RT-PCR in the intact pancreas and in human HEK 293 cells, isolated islets, and purified human beta-cells showed downregulation of DAF mRNA. mRNA was detectable in both of these preparations by RT-PCR; levels were lower following both the islet isolation process (53%) and single cell preparation (a further 62%) compared to HEK 293 controls. Human islet allotransplantation might be more successful if either de novo transfer of DAF onto the isolated islets or novel techniques for islet isolation preserving DAF could be developed.

Imaging islets labeled with magnetic nanoparticles at 1.5 Tesla.

We have developed a magnetic resonance imaging (MRI) technique for imaging Feridex (superparamagnetic iron oxide [SPIO])-labeled islets of Langerhans using a standard clinical 1.5-Tesla (T) scanner and employing steady-state acquisition imaging sequence (3DFIESTA). Both porcine and rat islets were labeled with SPIO by a transfection technique using a combination of poly-l-lysine and electroporation. Electron microscopy demonstrated presence of SPIO particles within the individual islet cells, including beta-cells and particles trapped between cell membranes. Our labeling method produced a transfection rate of 860 pg to 3.4 ng iron per islet, dependent on the size of the islet. The labeling procedure did not disrupt either the function or viability of the islets. In vitro 3DFIESTA magnetic resonance images of single-labeled islets corresponded with their optical images. In vivo T2*-weighted scan using 1.5 T detected as few as 200 SPIO-labeled islets transplanted under rat kidney capsule, which correlated with immunohistochemistry of the transplant for insulin and iron. Ex vivo 3DFIESTA images of kidneys containing 200, 800 or 2,000 SPIO-labeled islet isografts showed good correlation between signal loss and increasing numbers of islets. These data provide evidence that islets can be labeled with SPIO and imaged using clinically available 1.5- T MRI.

Assessment of acute antivascular effects of vandetanib with high-resolution dynamic contrast-enhanced computed tomographic imaging in a human colon tumor xenograft model in the nude rat.

Tumor size is not a reliable marker for the assessment of early antivascular effects of antiangiogenics. In the present study, we used 200-microm in-plane high-resolution dynamic contrast-enhanced computed tomography (DCE-CT) to noninvasively assess the immediate antivascular effects of vandetanib in a subcutaneous human colon cancer (LoVo) xenograft model in nude rats and to investigate correlation between changes in CT perfusion parameters and tumor volume or immunohistochemical end points. At 3 to 4 weeks after LoVo cell implantation, the animal was gavaged with either vandetanib (50 mg/kg) or vehicle twice (22 hours apart) and scanned with a preclinical DCE-CT scanner before (0 hour) and after treatment (24 hours). Quantitative maps of blood flow (BF) and volume (BV) of the tumor were calculated from the acquired DCE-CT images. The rats were divided into nonhypovascular, hypovascular, and combined (regardless of vascularity) groups. In the nonhypovascular group, significant decreases in both tumor BF and BV were observed in the vandetanib-treated rats compared with increases in the vehicle-treated rats. A significant decrease in BV was detected in the vandetanib-treated rats in the combined group as well. No differences in tumor growth, vascular endothelial growth factor expression, microvessel density, or apoptosis were observed between vandetanib- and vehicle-treated rats in all three groups. These results demonstrate that BF and BV imaging biomarkers from DCE-CT imaging can be used for rapid monitoring of immediate (24 hours after) antimicrovascular effects of vandetanib on tumors, even in the absence of significant changes of tumor volume or clinically relevant immunohistochemical end points.