Genetic variation in adiponectin (ADIPOQ) and the type 1 receptor (ADIPOR1), obesity and prostate cancer in African Americans.

Adiponectin is a protein derived from adipose tissue suspected to have an important role in prostate carcinogenesis. Variants in the adiponectin gene (ADIPOQ) and its type 1 receptor (ADIPOR1) have been recently linked to risk of both breast and colorectal cancer. Therefore, we set out to examine the relationship between polymorphisms in these genes, obesity and prostate cancer in study of African-American men. Ten single-nucleotide polymorphisms (SNPs) in ADIPOQ and ADIPOR1 were genotyped in DNA samples from 131 African-American prostate cancer cases and 344 controls participating in the Flint Men's Health Study. Logistic regression was then used to estimate their association with prostate cancer and obesity. While no significant associations were detected between any of the tested SNPs and prostate cancer, the rs1501299 SNP in ADIPOQ was significantly associated with body mass (P=0.03). Genetic variation in ADIPOQ and ADIPOR1 did not predict risk of prostate cancer in this study of African-American men. However, the rs1501299 SNP in ADIPOQ was associated with obesity. Further investigation is warranted to determine if racial differences exist in the influence of the adiponectin pathway on prostate cancer risk.

Absence of truncating BRIP1 mutations in chromosome 17q-linked hereditary prostate cancer families.

BACKGROUND: In a genome-wide scan (GWS) of 175 multiplex prostate cancer (PCa) families from the University of Michigan Prostate Cancer Genetics Project (PCGP), linkage was observed to markers on chromosome 17q21-24, a region that includes two breast cancer susceptibility genes, BRCA1 and BRIP1. BRIP1 is a Fanconi anaemia gene (FANCJ) that interacts with the BRCT domain of BRCA1 and has a role in DNA damage repair. Protein truncating mutations in BRIP1 have been identified in hereditary breast and ovarian cancer families, and a recent report suggested that a recurrent truncating mutation (R798X) may have a role in PCa susceptibility. METHODS: We examined the role of BRIP1 mutations in hereditary PCa through sequence analysis of 94 individuals from PCGP families showing linkage to 17q. RESULTS: A total of 24 single-nucleotide polymorphisms, including 7 missense variants but no protein truncating mutations, were observed. CONCLUSION: The data presented here suggest that BRIP1 truncating mutations are uncommon in PCa cases and do not account for the linkage to chromosome 17q observed in our GWS. Additional investigation is needed to determine the significance, if any, of the observed BRIP1 missense variants in hereditary PCa.

Functional rescue of the nephrogenic diabetes insipidus-causing vasopressin V2 receptor mutants G185C and R202C by a second site suppressor mutation.

Mutations in the gene of the G protein-coupled vasopressin V2 receptor (V2 receptor) cause X-linked nephrogenic diabetes insipidus (NDI). Most of the missense mutations on the extracellular face of the receptor introduce additional cysteine residues. Several groups have proposed that these residues might disrupt the conserved disulfide bond of the V2 receptor. To test this hypothesis, we first calculated a structure model of the extracellular receptor domains. The model suggests that the additional cysteine residues may form a second disulfide bond with the free, nonconserved extracellular cysteine residue Cys-195 rather than impairing the conserved bond. To address this question experimentally, we used the NDI-causing mutant receptors G185C and R202C. Their Cys-195 residues were replaced by alanine to eliminate the hypothetical second disulfide bonds. This second site mutation led to functional rescue of both NDI-causing mutant receptors, strongly suggesting that the second disulfide bonds are indeed formed. Furthermore we show that residue Cys-195, which is sensitive to "additional cysteine" mutations, is not conserved among the V2 receptors of other species and that the presence of an uneven number of extracellular cysteine residues, as in the human V2 receptor, is rare among class I G protein-coupled receptors.

The role of conserved extracellular cysteine residues in vasopressin V2 receptor function and properties of two naturally occurring mutant receptors with additional extracellular cysteine residues.

The G protein-coupled vasopressin V2 receptor (V2 receptor) contains a pair of conserved cysteine residues (C112 and C192) which are thought to form a disulfide bond between the first and second extracellular loops. The conserved cysteine residues were found to be important for the correct formation of the ligand binding domain of some G protein-coupled receptors. Here we have assessed the properties of the V2 receptor after site-directed mutagenesis of its conserved cysteine residues in transiently transfected human embryonic kidney (HEK 293) cells. Mutant receptors (C112S, C112A and C192S, C192A) were non-functional and located mostly in the cell's interior. The conserved cysteine residues of the V2 receptor are thus not only important for the structure of the ligand binding domain but also for efficient intracellular receptor transport. In addition to the functional significance of the conserved cysteine residues, we have also analyzed the defects of two mutant V2 receptors which cause X-linked nephrogenic diabetes insipidus (NDI) by the introduction of additional cysteine residues into the second extracellular loop (mutants G185C, R202C). These mutations are assumed to impair normal disulfide bond formation. Mutant receptor G185C and R202C were efficiently transported to the plasma membrane but were defective in ligand binding. Only in the case of the mutant receptor R202C, the more sensitive adenylyl cyclase activity assay revealed vasopressin-stimulated cAMP formation with a 35-fold increased EC(50) value and with a reduced EC(max), indicating that ligand binding is not completely abolished. Taking the unaffected intracellular transport of both NDI-causing mutant receptors into account, our results indicate that the observed impairment of ligand binding by the additional cysteine residues is not due to the prevention of disulfide bond formation between the conserved cysteine residues.

Comparisons of a proposed five-seed assay method with the single-seed and batch assay methods for I-125 seeds in ultrasound-guided prostate implants.

A simple five-seed assay method was proposed and investigated. A commercial well ion chamber system with an NIST-traceable single-seed calibration constant was used for the single-seed assays. A batch seed holder was used for batch assays. For the five-seed assays, a second single-seed holder was modified such that all five seeds were loaded in a central region of the well ion chamber. Compared with the same seed in the standard single-seed holder, the relative chamber responses for the five seed positions were 0.993, 0.993, 1.000, 1.001, and 0.977, respectively, indicating little or no position-dependent chamber response and no self-attenuation among seeds. Subsequent comparison of assays with the single-seed and five-seed methods indicated only 0.4% difference in charge collection. The five-seed calibration constant was therefore taken to be the same as the single-seed calibration constant. The reproducibility of the five-seed assay method was found to be better than 0.8%. When a dummy seed replaced an active seed, a nearly 20% reduction in charge was found, indicating that the proposed five-seed assay method can detect a dead seed. Clinical comparison of all three assay methods showed that they produced qualitatively the same assay results when the batch assay method was performed with extra care. Compared with the single-seed assay method, the five-seed method is equally simple, rigid, and reproducible, but it demands much less assay time. Compared with the batch assay method, the five-seed method is much more reproducible and reliable because of its rigid assay geometry; it only demands a moderate amount of assay time and can detect dead seeds. The American Association of Physicists in Medicine Task Group 40 (AAPM TG40) states that, for brachytherapy, ideally every (i.e., 100%) loose seed should be calibrated. For procedures involving large number of loose seeds, it then recommends that 10% of seeds be calibrated. The proposed five-seed assay is very simple to implement. It will facilitate the compliance of the "10%" recommendation from the AAPM TG40; it will make the "ideally 100%" statement from AAPM TG40 a more realistic and practical QA procedure in seed assaying.

Balloon dilatation and stent placement of suprahepatic caval anastomotic stenosis following liver transplantation.

We present an unusual case of suprahepatic caval anastomotic stenosis after adult liver transplantation in the early postoperative period. Color flow Doppler sonography and cavography were used to confirm the clinical diagnosis. Successful nonoperative treatment of the obstruction was achieved by balloon dilatation and subsequent implantation of a vascular stent.