Molecular basis of non-syndromic hypospadias: systematic mutation screening and genome-wide copy-number analysis of 62 patients.

STUDY QUESTION: What percentage of cases with non-syndromic hypospadias can be ascribed to mutations in known causative/candidate/susceptibility genes or submicroscopic copy-number variations (CNVs) in the genome? SUMMARY ANSWER: Monogenic and digenic mutations in known causative genes and cryptic CNVs account for >10% of cases with non-syndromic hypospadias. While known susceptibility polymorphisms appear to play a minor role in the development of this condition, further studies are required to validate this observation. WHAT IS KNOWN ALREADY: Fifteen causative, three candidate, and 14 susceptible genes, and a few submicroscopic CNVs have been implicated in non-syndromic hypospadias. STUDY DESIGN, SIZE, DURATION: Systematic mutation screening and genome-wide copy-number analysis of 62 patients. PARTICIPANTS/MATERIALS, SETTING, METHODS: The study group consisted of 57 Japanese and five Vietnamese patients with non-syndromic hypospadias. Systematic mutation screening was performed for 25 known causative/candidate/susceptibility genes using a next-generation sequencer. Functional consequences of nucleotide alterations were assessed by in silico assays. The frequencies of polymorphisms in the patient group were compared with those in the male general population. CNVs were analyzed by array-based comparative genomic hybridization and characterized by fluorescence in situ hybridization. MAIN RESULTS AND THE ROLE OF CHANCE: Seven of 62 patients with anterior or posterior hypospadias carried putative pathogenic mutations, such as hemizygous mutations in AR, a heterozygous mutation in BNC2, and homozygous mutations in SRD5A2 and HSD3B2. Two of the seven patients had mutations in multiple genes. We did not find any rare polymorphisms that were abundant specifically in the patient group. One patient carried mosaic dicentric Y chromosome. LIMITATIONS, REASONS FOR CAUTION: The patient group consisted solely of Japanese and Vietnamese individuals and clinical and hormonal information of the patients remained rather fragmentary. In addition, mutation analysis focused on protein-altering substitutions. WIDER IMPLICATIONS OF THE FINDINGS: Our data provide evidence that pathogenic mutations can underlie both mild and severe hypospadias and that HSD3B2 mutations cause non-syndromic hypospadias as a sole clinical manifestation. Most importantly, this is the first report documenting possible oligogenicity of non-syndromic hypospadias. STUDY FUNDING/COMPETING INTERESTS: This study was funded by the Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology; by the Grant-in-Aid from the Japan Society for the Promotion of Science; by the Grants from the Ministry of Health, Labour and Welfare, from the National Center for Child Health and Development and from the Takeda Foundation. The authors have no competing interests to disclose. TRIAL REGISTRATION NUMBER: Not applicable.

HapMap SNP Scanner: an online program to mine SNPs responsible for cell phenotype.

Minor histocompatibility (H) antigens are targets of graft-vs-host disease and graft-vs-tumor responses after human leukocyte antigen matched allogeneic hematopoietic stem cell transplantation. Recently, we reported a strategy for genetic mapping of linkage disequilibrium blocks that encoded novel minor H antigens using the large dataset from the International HapMap Project combined with conventional immunologic assays to assess recognition of HapMap B-lymphoid cell line by minor H antigen-specific T cells. In this study, we have constructed and provide an online interactive program and demonstrate its utility for searching for single-nucleotide polymorphisms (SNPs) responsible for minor H antigen generation. The website is available as 'HapMap SNP Scanner', and can incorporate T-cell recognition and other data with genotyping datasets from CEU, JPT, CHB, and YRI to provide a list of candidate SNPs that correlate with observed phenotypes. This method should substantially facilitate discovery of novel SNPs responsible for minor H antigens and be applicable for assaying of other specific cell phenotypes (e.g. drug sensitivity) to identify individuals who may benefit from SNP-based customized therapies.

[Clinical study on chlormadinone acetate alone followed by combination with LH-RH analogue for prostatic cancer: effects on lipid metabolism].

Twenty-four previously untreated patients with a diagnosis of prostatic cancer were treated with chlormadinone acetate (CMA) alone (100 mg/day) for 4 weeks, and luteinizing hormone-releasing hormone analogue (LH-RHa) was added for the next 24 weeks. Marked decreases in blood LH, testosterone (T), prostate specific antigen (PSA), gamma-seminoprotein (gamma-Sm), and prostatic acid phosphatase (PAP) were observed after a single dose of CMA. T levels were significantly increased 3 days after the initial dose of LH-RHa, and did not return to the pretreatment level. There were no significant increases in any of the markers, nor were there any flare-up cases. Triglyceride levels, which were slightly elevated before the start of treatment, were significantly decreased 24 weeks after the completion of combined therapy. PSA was evaluated as partial response (PR) or better in 86.7% of the patients. Overall evaluation showed PR or better in 75.0% of the patients. These findings suggest that prior administration of CMA followed by combined administration with LH-RHa is useful in the treatment of prostatic cancer. No negative effects on lipid metabolism were observed at any time during the treatment period.

The effect of renal transplantation on a major endogenous ligand retained in uremic serum.

The effects of renal transplantation on serum concentrations of 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF) and indole-3-acetic acid (IAA), which are endogenous ligands retained in uremic serum, and on phenytoin binding to serum protein were investigated. IAA, a weakly bound ligand, was rapidly excreted by the transplanted kidney during the first one to three days after renal transplantation, but CMPF, a strongly bound ligand, was slowly excreted. The binding defect of phenytoin was partially corrected by transplantation during the period of study. The results suggested that the prolonged drug binding defect observed despite successful renal transplantation is caused by a slower decrease of strongly bound ligands such as CMPF retained in uremic serum; hypoalbuminemia, usually observed after transplantation, may also contribute to this phenomenon.

Infiltrating ductal carcinoma developing within cystosarcoma phyllodes--a case report.

Malignancy in cystosarcoma phyllodes is uncommon and often confined to the stromal element. An extremely rare case of infiltrating ductal carcinoma developing within the stroma of cystosarcoma phyllodes is reported herein. A breast tumor with a diameter of 15 cm, which was diagnosed as cystosarcoma phyllodes, was excised from the right breast of a 47-year old woman. The histopathological examination revealed that hyperplastic ductal epithelial cells with dark cytoplasm and enlarged hyperchromatic nuclei were infiltrating the stroma. Thus, a diagnosis of ductal carcinoma within cystosarcoma was made. Subsequently, a standard radical mastectomy was performed. No recurrence or metastasis has been observed over the post-operative period of 5 years and 6 months.

The successful outcome of second kidney transplantation and its contributing factors.

Fourteen out of 301 patients who underwent allogeneic kidney transplantations, between April, 1970 and December, 1987, received second kidney allografts, including 4 living and 10 cadaveric grafts. The survival of the second grafts transplanted in those 14 recipients was superior to that of the first grafts transplanted in the other 287 recipients. Furthermore, the survival of the second grafts from 4 years onwards was significantly higher than that of the first grafts, in spite of a higher population of cadaveric grafts used in the second transplantation than in the first. Although it was impossible to determine the main factor which induced the improved survival of the second grafts when compared with that of the first, a combination of beneficial factors, such as a high rate of living related transplantation resulting in long-term graft survival of the first transplantation, the administration of immunosuppressive drugs during the period of re-hemodialysis and blood transfusion prior to the second transplantation, was considered to be the reason why successful second graft survival was achieved.

Results of kidney transplantation in 25 pediatric patients.

Outcome, long-term prognosis, growth activity and rehabilitation after kidney transplantation were studied in 25 pediatric patients transplanted with a kidney graft from one-haplotype identical parent. Excellent patient and graft survival with low incidences of acute rejection or serious complications could be achieved in this population, as compared with the results of adult recipients. Growth retardations in height and weight were observed in these patients before transplantation, and were significantly correlated with the duration of low or no kidney function. In 12 recipients who were transplanted at ages of younger than 15 years and followed up over two years, a dramatic increase in weight appeared within one year after transplantation and a greater increase in height was exhibited in the second and third year than in the first. Increase in height was significantly greater in those children transplanted at ages of younger than 10 years than in those transplanted at ages of older than 11 years. Catch-up growth was observed in one-third of these children. Retrospectively, there was no difference in the doses of prednisolone given between the two groups of patients with, and without catch-up growth, but the incidence of acute rejection was higher in the group without catch-up growth. Currently, 18 recipients have functioning grafts and 16 (88.9 per cent) of them are in full-time school or working. From these results it is concluded that kidney transplantation is the first feasible manoeuvre for those children with chronic renal failure and it should be performed as soon an possible in order to preserve their growth activity.