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Small farm producers' sustenance depends on their alpaca herds and the production of fiber. Genetic improvement of fiber characteristics would increase their economic benefits and quality of life. The incorporation of molecular marker technology could overcome current limitations for the implementation of genetic improvement programs. Hence, the aim of this project was the generation of an alpaca single nucleotide polymorphism (SNP) microarray. A sample of 150 Huacaya alpacas from four farms, two each in Puno and Cerro de Pasco were used for SNP discovery by genotyping by sequencing (GBS). Reduced representation libraries, two per animal, were produced after DNA digestion with ApeK1 and double digestion with Pst1-Msp1. Ten alpaca genomes, sequenced at depths between 12× to 30×, and the VicPac3.1 reference genome were used for read alignments. Bioinformatics analysis discovered 76,508 SNPs included in the microarray. Candidate genes SNPs (302) for fiber quality and color are also included. The microarray SNPs cover 90.5% of the genome length with a density of about 39 ± 2.51 SNPs/Mb of DNA at an average interval of 26.45 ± 18.57 kbp. The performance was evaluated by genotyping 30 family trios and comparing them to their pedigrees, as well as comparing microarray to GBS genotypes. Concordance values of 0.93 and 0.94 for ApeK1 and Pst1-Msp1 generated SNPs were observed. Similarly, 290 fiber quality and color candidate gene SNPs were validated. Availability of this microarray will facilitate genome-wide association studies, marker-assisted selection and, in time, genomic selection.

Alpaca is a camelid species of broad economic, biological and biomedical interest, and an essential part of the cultural and historical heritage of Peru. Recently, efforts have been made to improve knowledge of the alpaca genome, and its genetics and cytogenetics, to develop molecular tools for selection and breeding. Here, we report cytogenetic mapping of 35 new markers to 19 alpaca autosomes and the X chromosome. Twenty-eight markers represent alpaca SNPs, of which 17 are located inside or near protein-coding genes, two are in ncRNA genes and nine are intergenic. The remaining seven markers correspond to candidate genes for fiber characteristics ( BMP4, COL1A2, GLI1, SFRP4) , coat color ( TYR ) and development ( CHD7, PAX7 ). The results take the tally of cytogenetically mapped markers in alpaca to 281, covering all 36 autosomes and the sex chromosomes. The new map assignments overall agree with human-camelid conserved synteny data, except for mapping BMP4 to VPA3, suggesting a hitherto unknown homology with HSA14. The findings validate, refine and correct the current alpaca assembly VicPac3.1 by anchoring unassigned sequence scaffolds, and ordering and orienting assigned scaffolds. The study contributes to the improvement in the alpaca reference genome and advances camelid molecular cytogenetics.

Objective: To evaluate the sensitivity and clinical utility of intraoperative mobile gamma camera (MGC) imaging in sentinel lymph node biopsy (SLNB) in melanoma., Background: The false-negative rate for SLNB for melanoma is approximately 17%, for which failure to identify the sentinel lymph node (SLN) is a major cause. Intraoperative imaging may aid in detection of SLN near the primary site, in ambiguous locations, and after excision of each SLN. The present pilot study reports outcomes with a prototype MGC designed for rapid intraoperative image acquisition. We hypothesized that intraoperative use of the MGC would be feasible and that sensitivity would be at least 90%., Methods: From April to September 2008, 20 patients underwent Tc99 sulfur colloid lymphoscintigraphy, and SLNB was performed with use of a conventional fixed gamma camera (FGC), and gamma probe followed by intraoperative MGC imaging. Sensitivity was calculated for each detection method. Intraoperative logistical challenges were scored. Cases in which MGC provided clinical benefit were recorded., Results: Sensitivity for detecting SLN basins was 97% for the FGC and 90% for the MGC. A total of 46 SLN were identified: 32 (70%) were identified as distinct hot spots by preoperative FGC imaging, 31 (67%) by preoperative MGC imaging, and 43 (93%) by MGC imaging pre- or intraoperatively. The gamma probe identified 44 (96%) independent of MGC imaging. The MGC provided defined clinical benefit as an addition to standard practice in 5 (25%) of 20 patients. Mean score for MGC logistic feasibility was 2 on a scale of 1-9 (1 = best)., Conclusions: Intraoperative MGC imaging provides additional information when standard techniques fail or are ambiguous. Sensitivity is 90% and can be increased. This pilot study has identified ways to improve the usefulness of an MGC for intraoperative imaging, which holds promise for reducing false negatives of SLNB for melanoma.

Optimization of exposure parameters (target, filter, and kVp) in digital mammography necessitates maximization of the image signal-to-noise ratio (SNR), while simultaneously minimizing patient dose. The goal of this study is to compare, for each of the major commercially available full field digital mammography (FFDM) systems, the impact of the selection of technique factors on image SNR and radiation dose for a range of breast thickness and tissue types. This phantom study is an update of a previous investigation and includes measurements on recent versions of two of the FFDM systems discussed in that article, as well as on three FFDM systems not available at that time. The five commercial FFDM systems tested, the Senographe 2000D from GE Healthcare, the Mammomat Novation DR from Siemens, the Selenia from Hologic, the Fischer Senoscan, and Fuji's 5000MA used with a Lorad M-IV mammography unit, are located at five different university test sites. Performance was assessed using all available x-ray target and filter combinations and nine different phantom types (three compressed thicknesses and three tissue composition types). Each phantom type was also imaged using the automatic exposure control (AEC) of each system to identify the exposure parameters used under automated image acquisition. The figure of merit (FOM) used to compare technique factors is the ratio of the square of the image SNR to the mean glandular dose. The results show that, for a given target/filter combination, in general FOM is a slowly changing function of kVp, with stronger dependence on the choice of target/filter combination. In all cases the FOM was a decreasing function of kVp at the top of the available range of kVp settings, indicating that higher tube voltages would produce no further performance improvement. For a given phantom type, the exposure parameter set resulting in the highest FOM value was system specific, depending on both the set of available target/filter combinations, and on the receptor type. In most cases, the AECs of the FFDM systems successfully identified exposure parameters resulting in FOM values near the maximum ones, however, there were several examples where AEC performance could be improved.

Breast scintigraphy is a technique by which the biological properties of breast lesions can be assessed using an injected radiopharmaceutical. It may be particularly useful for women with radiographically dense breasts, in whose mammograms, lesions are often obscured by breast tissue. We are evaluating a dual modality breast scanner developed at the University of Virginia for its ability to distinguish between benign and malignant lesions. The scanner obtains a digital mammogram and a gamma ray emission image in quick succession with the breast held under mild compression, resulting in a fused image in which structures in the digital mammogram can be directly correlated with those in the scintigram. Our experience has shown that radiopharmaceutical uptake by normal breast tissue can sometimes obscure uptake by small lesions. It would therefore be advantageous to correct for this background uptake if possible. One potential way of accomplishing this is to use the information from the digital mammogram to help predict the background radiopharmaceutical distribution. With this in mind, we retrospectively investigated the degree of spatial correlation between the distribution of background activity and the distribution of radiodense breast tissue in normal breasts. Using a histogram-based analysis, we have quantified the degree of correlation in 16 images obtained from a total of 8 patients. We also used the mammographic images to quantify the radiographic density of each breast. Our results suggest that spatial correlation between areas of high radiopharmaceutical uptake and parenchymal density exists in the most dense regions of the breast for either extremely dense or heterogeneously dense breasts. High correlation was also observed for some homogeneously fatty breasts. In the latter case however, variation in breast thickness appeared to be the cause of the increased correlation. Correlation properties are approximately equal in both right and left breasts for a particular patient, except in cases exhibiting focal radiotracer uptake in a lesion. Although our preliminary results suggest that correlation between radiopharmaceutical uptake and parenchymal density exists, the number of cases thus far is too small for definitive conclusions. In addition, the planar nature of the dual modality scans imposes an inherent limitation on our ability to take into account attenuation of the emitted gamma radiation, which thus constitutes an uncontrolled variable in the correlation analysis. In principle, this problem can be eliminated by 3-dimensional imaging.

Scintimammography, or single gamma nuclear imaging of the breast, has shown promise as a way of characterizing certain biological properties of suspicious breast masses. Conventional scintimammography, performed using large clinical gamma cameras and prone patient positioning suffers from several drawbacks including poor sensitivity for small (> 1 cm) lesions and no reliable method for correlating scintigraphic findings with those of other imaging modalities. We are developing a system designed to overcome some of these problems. The system combines x-ray mammography with scintimammography on a common gantry. The x-ray and gamma ray images are obtained in quick succession, with the breast in a common configuration under mild compression. A digital x-ray detector is used, permitting rapid assessment of lesion location prior to gamma imaging, and enabling fusion of the x-ray transmission and gamma emission information in a single digital image. In a pilot clinical diagnostic study, the system has demonstrated high pathology-proven accuracy in differentiating benign and malignant masses.