Segmentation of the prostate from suprapubic ultrasound images

We present a technique for semiautomated segmentation of human prostates using suprapubic ultrasound (US) images. In this approach, a speckle reducing anisotropic diffusion (SRAD) is applied to enhance the images and the instantaneous coefficient of variation (ICOV) is utilized for edge detection. Segmentation is accomplished via a parametric active contour model in a polar coordinate system that is tailored to the application. The algorithm initially approximates the prostate boundary in two stages. First a primary contour is detected using an elliptical model, followed by a primary contour optimization using an area-weighted mean-difference binary flow geometric snake model. The algorithm was assessed by comparing the computer-derived contours with contours produced manually by three sonographers. The proposed method has application in radiation therapy planning and delivery, as well as in automated volume measurements for ultrasonic diagnosis. The average root mean square discrepancy between computed and manual outlines is less than the inter-observer variability. Furthermore, 76% of the computer-outlined contour is less than 1 sigma manual outline variance away from "true" boundary of prostate. We conclude that the methods developed herein possess acceptable agreement with manually contoured prostate boundaries and that they are potentially valuable tools for radiotherapy treatment planning and verification.     

Construction of a high resolution linkage disequilibrium map to evaluate common genetic variation in TP53 and neural tube defect risk in an Irish population

Genetic and environmental factors contribute to the etiology of neural tube defects (NTDs). While periconceptional folic acid supplementation is known to significantly reduce the risk of NTDs, folate metabolic pathway related factors do not account for all NTDs. Evidence from mouse models indicates that the tumor protein p53 (TP53) is involved in implantation and normal neural tube development. To determine whether genetic variation in TP53 might contribute to NTD risk in humans, we constructed a high resolution linkage disequilibrium (LD) map of the TP53 genomic region based on genotyping 21 markers in an Irish population. We found that nine of these variants can be used to capture the majority of common variation in the TP53 genomic region. In contrast, the 3-marker haplotype commonly reported in the TP53 literature offers limited coverage of the variation in the gene. We used the expanded set of polymorphisms to measure the influence of TP53 on NTDs using both case-control and family based tests of association. We also assayed a functional variant in the p53 regulator MDM2 (rs2279744). Alleles of three noncoding TP53 markers were associated with NTD risk. A case effect was seen with the GG genotype of rs1625895 in intron 6 (OR = 1.37 [1.04-1.79], P = 0.02). A maternal effect was seen with the 135/135 genotype of the intron 1 VNTR (OR = 1.86 [1.16-2.96], P = 0.01) and the TT genotype of rs1614984 (RR = 0.58 [0.37-0.91], P = 0.02). As multiple comparisons were made, these cannot be considered definitive positive findings and additional investigation is required.     

Evaluation of common genetic variants in 82 candidate genes as risk factors for neural tube defects

ABSTRACT: BACKGROUND: Neural tube defects (NTDs) are common birth defects (~1 in 1000 pregnancies in the US and Europe) that have complex origins, including environmental and genetic factors. A low level of maternal folate is one well-established risk factor, with maternal periconceptional folic acid supplementation reducing the occurrence of NTD pregnancies by 50-70%. Gene variants in the folate metabolic pathway (e.g., MTHFR rs1801133 (677 C > T) and MTHFD1 rs2236225 (R653Q)) have been found to increase NTD risk. We hypothesized that variants in additional folate/B12 pathway genes contribute to NTD risk. METHODS: A tagSNP approach was used to screen common variation in 82 candidate genes selected from the folate/B12 pathway and NTD mouse models. We initially genotyped polymorphisms in 320 Irish triads (NTD cases and their parents), including 301 cases and 341 Irish controls to perform case-control and family based association tests. Significantly associated polymorphisms were genotyped in a secondary set of 250 families that included 229 cases and 658 controls. The combined results for 1441 SNPs were used in a joint analysis to test for case and maternal effects. RESULTS: Nearly 70 SNPs in 30 genes were found to be associated with NTDs at the p < 0.01 level. The ten strongest association signals (p-value range: 0.0003-0.0023) were found in nine genes (MFTC, CDKN2A, ADA, PEMT, CUBN, GART, DNMT3A, MTHFD1 and T (Brachyury)) and included the known NTD risk factor MTHFD1 R653Q (rs2236225). The single strongest signal was observed in a new candidate, MFTC rs17803441 (OR = 1.61 [1.23-2.08], p = 0.0003 for the minor allele). Though nominally significant, these associations did not remain significant after correction for multiple hypothesis testing. CONCLUSIONS: To our knowledge, with respect to sample size and scope of evaluation of candidate polymorphisms, this is the largest NTD genetic association study reported to date. The scale of the study and the stringency of correction are likely to have contributed to real associations failing to survive correction. We have produced a ranked list of variants with the strongest association signals. Variants in the highest rank of associations are likely to include true associations and should be high priority candidates for further study of NTD risk.     

Genetic analysis of the 3' untranslated region of the tumour necrosis factor shows a highly conserved region in rheumatoid arthritis affected and unaffected subjects

Tumour necrosis factor (TNF) is a key proinflammatory mediator in rheumatoid arthritis (RA). The TNF locus, situated in the class III region of the MHC, is flanked by five microsatellite markers. It has previously been shown that this region influences susceptibility to RA; two TNF microsatellite haplotypes were found to be associated with RA. Evidence from murine studies has indicated that variation in the TNF 3' untranslated region (UTR) could be associated with altered regulation of TNF biosynthesis. In order to identify possible RA associated polymorphisms, more than 800 bp of the TNF 3' UTR was genetically analysed in RA affected and unaffected subjects possessing specific RA and non-RA associated TNF microsatellite haplotypes. The TNF 3' UTR region was analysed using two mutation detection methods, PCR-SSCP and NIRCA analysis and DNA sequencing. No genetic differences were observed in the human TNF 3' UTR between subjects, that is, irrespective of RA status or TNF haplotype, and also compared with previously published TNF sequences from human sources. Therefore it can be concluded that the TNF 3' UTR in this population was highly conserved and did not influence susceptibility to RA.     

Treatment planning considerations and quality assurance for CT-guided transischiorectal implantation of the prostate.

A CT-guided technique for prostate brachytherapy has been developed which can be performed on patients with large prostates and which allows real time evaluation and modification of implant geometry. The patient is positioned prone and radioactive Pd 103 or I 125 seeds are implanted through the transischiorectal space. Needles are inserted through a template whose plane is oriented at a nominal angle of 26 degrees away from the horizontal to facilitate needle penetration through the ischiorectal space. The CT gantry is tilted 26 degrees, so that its plane is orthogonal to that of the template. The oblique geometry between the CT gantry and direction of couch translation must be considered during source position planning, implantation, and post-treatment evaluation. This consideration is presented along with discussion of relevant quality assurance procedures and recommended tolerances. The mechanical and radiological tolerances of the CT scanner must be consistent with the high level of precision required in radiation therapy. Special emphasis was placed on gantry laser and image plane alignment, sensitivity and radiation profiles, and spatial accuracy of image reconstruction, table translation, and gantry tilt.     

Glomus tympanicum chemodectomas: radiographic and clinical characteristics

Glomus tympanicum chemodectomas are benign neoplasms that develop from normal glomus bodies located along the Jacobson (tympanic) nerve in the middle ear. The medical charts and radiographic studies of 55 patients with these tumors were reviewed. Women outnumbered men in a ratio of 3.5:1, and the patients' average age when they initially reported symptoms was 52 years. Tinnitus, ear pulsations, and diminished hearing were the most frequent symptoms. No patient had a second chemodectoma, and none of seven patients who were tested had elevated neuroendocrine compounds. Review of the radiographic examinations showed that direct coronal, thin-section computed tomography (CT) was the most sensitive means of demonstrating glomus tympanicum chemodectomas. Magnification angiography was also a sensitive diagnostic study, typically depicting a trapezoidal, hypervascular, middle-ear mass that appeared initially in the middle-to-late arterial phase and quickly disappeared in the venous phase. Differentiation from an aberrant internal carotid artery is critical to prevent arterial biopsy.     

Human erythrocyte antigens: II. The In(Lu) gene regulates expression of an antigen on an 80-kilodalton protein of human erythrocytes

We have previously shown that a murine monoclonal antibody (A3D8) identifies a human erythrocyte protein antigen whose expression is regulated by the Lutheran inhibitor [In(Lu)] gene. In the present study, we demonstrated by immunoprecipitation and Western blot techniques that the antigen defined by A3D8 was on an 80-kD erythrocyte membrane protein. A second 170-kD protein was coprecipitated with the 80-kD protein but failed to show antigen activity by Western blot analysis. The 170-kD protein, when analyzed by sodium dodecyl sulfate- polyacrylamide gel electrophoresis in two dimensions, was composed of 50- and 30-kD disulfide-linked subunits. In(Lu) Lu[a-b-) erythrocytes differed from Lu(a+b+) or Lu(a-b+) erythrocytes in that In(Lu) deoxycholate erythrocyte membrane extracts contained trace amounts of immunoprecipitable 80-kD protein compared with detergent-solubilized erythrocyte membrane extracts prepared from Lu(a+b+) or Lu(a-b+) subjects.     

The effects of magnesium on calcium inhibition of parathyroid adenylate cyclase

cAMP has been shown to mediate the response of the parathyroid gland to a number of agonists and appears to take part in the regulation of this gland by divalent cations as well. We have studied the effects of the concentrations of free magnesium (Mg+2) and ionic calcium (Ca+2) on the kinetic properties of normal porcine parathyroid adenylate cyclase. In a previous study we obtained evidence for two calcium inhibition sites in this enzyme complex. In the present study we observed that the Mg+2 concentration influences the relative contribution of these sites to the overall calcium inhibition. At a high Mg+2 concentration (10 mM), the high affinity site contributes less than 50% of the total calcium inhibitable activity, whereas at a Mg+2 concentration in the low physiological range (0.5 mM), the high affinity site accounts for all the calcium inhibitable activity. Mg+2 was found to be a potent activator of porcine parathyroid adenylate cyclase, with a Ka of Mg of 0.8-2 mM. Ionic calcium at low concentrations (0.2-5 microM) acts as a competitive inhibitor with respect to Mg+2 activation. The calcium inhibition constant was estimated to be 2-3 microM. The Km for ATPMg-2 was 0.3 mM, which is similar to that found in other studies of adenylate cyclase activity in parathyroid tissue. The effects of Ca+2 on enzymatic activity with respect to the ATPMg-2 concentration showed noncompetitive inhibition. The calcium inhibition constant with respect to its effect on Vmax (KIv) was 3 microM; the calcium inhibition constant with respect to its effect on the binding of ATPMg-2 (KIs) was 10 microM. It is concluded from these results that the concentrations of intracellular Ca+2 reported to be present in parathyroid cells could inhibit adenylate cyclase activity. The mode of calcium inhibition that involves competition with magnesium would be particularly significant at low intracellular Mg+2 concentrations, and this phenomenon may account for the parathyroid secretory defect which is a characteristic feature of the magnesium-deficient state.     

The formation of Ca++-dependent complexes of platelet membrane glycoproteins IIb and IIIa in solution as determined by crossed immunoelectrophoresis

Triton X-100 soluble proteins from 125I-labeled human platelets were studied by crossed immunoelectrophoresis employing a multispecific rabbit antibody raised against whole normal platelets. Emphasis was placed upon an analysis of immunoprecipitates containing 125I-labeled major membrane glycoproteins, and in particular, a prominent immunoprecipitate containing a glycoprotein antigen (s) previously designated as protein 16. SDS-polyacrylamide gel electrophoresis of protein 16 precipitated by a monospecific alloantibody. IgG L . . . , confirmed the presence of both glycoproteins IIb and IIIa. 125I-IgG L . . . , at concentration below that capable of precipitating protein 16 by itself, bound specifically to the precipitate containing protein 16 produced by the multispecific rabbit antibody. No other precipitates formed by the rabbit antibody contained either glycoprotein IIb or IIIa. When platelet proteins, incubated with optimum concentrations of ethylenediamine tetraacetic acid (EDTA) or ethyleneglycol bis (B- aminoethylether) NN1-tetraacetic acid (EGTA), were electrophoresed against the rabbit antibody, previously unobserved immunoprecipitates that contained either free glycoprotein IIb or free glycoprotein IIIa were detected. Upon readdition of excess Ca++, but not Mg++, to the same protein samples, a single immunoprecipitate containing both glycoproteins was once again observed. It is thus demonstrated that glycoproteins IIb and IIIa can form Ca++-dependent complexes (protein 16) in Triton X-100 extracts of normal platelets. The potential significance of the reversible association of these glycoproteins to normal platelet function is discussed.