The authors present the results of accuracy measurements, obtained in both laboratory phantom studies and an in vivo assessment, for a technique of frameless stereotaxy. An instrument holder was developed to facilitate stereotactic guidance and enable introduction of frameless methods to traditional frame-based procedures. The accuracy of frameless stereotaxy was assessed for images acquired using 0.5-tesla or 1.5-tesla magnetic resonance (MR) imaging or 2-mm axial, 3-mm axial, or 3-mm helical computerized tomography (CT) scanning. A clinical series is reported in which biopsy samples were obtained using a frameless stereotactic procedure, and the accuracy of these procedures was assessed using postoperative MR images and image fusion. The overall mean error of phantom frameless stereotaxy was found to be 1.3 mm (standard deviation [SD] 0.6 mm). The mean error for CT-directed frameless stereotaxy was 1.1 mm (SD 0.5 mm) and that for MR image-directed procedures was 1.4 mm (SD 0.7 mm). The CT-guided frameless stereotaxy was significantly more accurate than MR image-directed stereotaxy (p = 0.0001). In addition, 2-mm axial CT-guided stereotaxy was significantly more accurate than 3-mm axial CT-guided stereotaxy (p = 0.025). In the clinical series of 21 frameless stereotactically obtained biopsies, all specimens yielded the appropriate diagnosis and no complications ensued. Early postoperative MR images were obtained in 16 of these cases and displacement of the biopsy site from the intraoperative target was determined by fusion of pre- and postoperative image data sets. The mean in vivo linear error of frameless stereotactic biopsy sampling was 2.3 mm (SD 1.9 mm). The mean in vivo Euclidean error was 4.8 mm (SD 2 mm). The implications of these accuracy measurements and of error in stereotaxy are discussed.

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Human amniotic fluid contains cells that potentially have important stem cell characteristics, yet the programs controlling their developmental potency are unclear. Here, we provide evidence that amniocytes derived from multiple patients are marked by heterogeneity and variability in expression levels of pluripotency markers. Clonal analysis from multiple patients indicates that amniocytes have large pools of self-renewing cells that have an inherent property to give rise to a distinct amniocyte phenotype with a heterogeneity of pluripotent markers. Significant to their therapeutic potential, genome-wide profiles are distinct at different gestational ages and times in culture, but do not differ between genders. Based on hierarchical clustering and differential expression analyses of the entire transcriptome, amniocytes express canonical regulators associated with pluripotency and stem cell repression. Their profiles are distinct from human embryonic stem cells (ESCs), induced-pluripotent stem cells (iPSCs), and newborn foreskin fibroblasts. Amniocytes have a complex molecular signature, coexpressing trophoblastic, ectodermal, mesodermal, and endodermal cell-type-specific regulators. In contrast to the current view of the ground state of stem cells, ESCs and iPSCs also express high levels of a wide range of cell-type-specific regulators. The coexpression of multilineage differentiation markers combined with the strong expression of a subset of ES cell repressors in amniocytes suggests that these cells have a distinct phenotype that is unlike any other known cell-type or lineage.

A polymerase chain reaction (PCR) method developed to detect varicella zoster virus (VZV) in clinical samples is based on amplifying sequences of viral gene 31, the coding region for glycoprotein II. Its sensitivity was evaluated by amplification from plasmid VZV DNA containing VZV gene 31; 45 copies were detected in 1 microgram of human DNA. In testing within 24 h after the onset of varicella exanthem, 21 (75%) of 28 lesion samples were positive by VZV PCR, whereas VZV was isolated from only 21% by a standard tissue culture method. Only 1 (3.3%) of 30 samples of oropharyngeal secretions but peripheral blood mononuclear cells from 8 (67%) of 12 patients were positive. The sensitivity and specificity of the VZV PCR method indicates its usefulness for investigating the pathogenesis of VZV infection. Direct contact with cutaneous lesions rather than respiratory secretions may be the most important route of VZV transmission from healthy individuals with acute varicella.