Prenatal diagnosis of a rare variant of hypospadias and review of the literature.

We present a case of a variant of hypospadias, diagnosed prenatally at 21 weeks' gestation, in which the penis and scrotum appeared normal but there was a cyst-like lesion and a urethrocutaneous fistula on the ventral side of the penis. Detailed sonographic examination and karyotyping confirmed this was an isolated lesion and helped the parents to decide on continuation of the pregnancy.

Umbilical cord blood transplantation for Maroteaux-Lamy syndrome (mucopolysaccharidosis type VI).

Severe Maroteaux-Lamy syndrome (mucopolysaccharidosis type VI) is usually fatal by early adulthood. Bone marrow transplantation is the only form of definitive enzyme replacement therapy available. A 5-year-old boy with Maroteaux-Lamy syndrome has successful recovery of bone marrow and enzymatic functions after umbilical cord blood transplant from his unaffected HLA-identical brother. Busulphan (16 mg/kg) and cyclophosphamide (200 mg/kg) were used as preparative chemotherapy with short methotrexate and long cyclosporin as prophylaxis against graft-versus-host disease (GVHD). A total of 6.08 x 10(7)/kg nucleated cells and 2.92 x 10(5)/kg CD34+ cells were transplanted with neutrophil engraftment achieved on day 26. There was no evidence of acute and chronic GVHD. Fifteen months after transplant, a normal level of N-acetylgalactosamine-4-sulphatase activity was achieved despite mixed chimerism. There was clinical improvement of hepatosplenomegaly, facial and skin features, joint mobility and resolution of suppurative middle ear effusion. He returned to school and continued to perform well in academic studies. We report here the first successful umbilical cord blood transplant as treatment of Maroteaux-Lamy syndrome.

Up-regulation of cell growth associated with an extra Y chromosome in a child with beta-thalassemia major having undergone hematopoietic stem cell transplant.

An extra Y chromosome(s) is occasionally found in patients with various hematologic neoplasias; however, an association with hereditary blood diseases is unknown. In a child with beta-thalassemia who received a transplant with sex-mismatched umbilical cord blood and neonatal blood, fluorescent in situ hybridization (FISH) with probes to X and Y chromosomes revealed an extra Y signal in 19.3% of the hepatocytes and in 38.9% of the pretransplant peripheral blood cells, yielding YY/Y ratios of 0.24 and 0.64, respectively. In granulocyte colony-stimulating factor-mobilized autologous peripheral blood stem cells, the FISH ratio of interphase XYY cells to XY cells was 1.64, and there were 3.4 times more G-banded XYY metaphases than normal metaphases. Both FISH and polymerase chain reaction persistently demonstrated posttransplant mixed chimerism. There was a greater proportion of residual autologous XYY cells than XY cells with a mean posttransplant YY/Y ratio of 1.56 (n = 13) (1 SD = 0.33; range, 1.10 to 2.17). In vitro clonogenic assays yielded a normal number of colony-forming units but no growth in cultures without growth factor supplement. This study suggests that hematopoietic stem cells with an extra Y chromosome may upregulate cell growth in response to cytokine stimulation. A posttransplant preponderance of XYY cells might be attributable to an extra Y chromosome in hematopoietic stem cells withstanding the myeloablative conditioning regimen.

Characterization of Ca(2+) flows essential in ornithine decarboxylase induction by L-asparagine in rat hepatoma cells using Ca(2+) flow inhibitors.

L-Asparagine stimulates bi-directional Ca(2+) flows and induces ornithine decarboxylase in Reuber H-35 hepatoma cells. Previously it has been shown that these effects are completely, but reversibly inhibited by lanthanum chloride. In this study we examined the role(s) of Ca(2+) flows using more specific Ca(2+) flow inhibitors. It was shown that ornithine decarboxylase induction was inhibited by CdCl(2) and verapamil at concentrations above 1 mu M and 100 mu M respectively, but was unaffected by as much as 300 mu M NiCl(2), 1 mM nifedipine, or 10 mu M omega-conotoxin. Enzyme induction was blocked by the Ca(2+)-ATPase pump antagonists vanadate and Compound 48/80 in a dose-dependent manner. These results, taken together with the observations that extracellular Ca(2+) is essential for enzyme induction but a substantial elevation of cytoplasmic [Ca(2+)] is not, suggest that Ca(2+) inflow independent of the receptor-activated Ca(2+) channels, and the Ca(2+)-ATPase mediated Ca(2+) out-flow, are both important factors in the action of L-asparagine.

[Studies on mode of human infection with Echinochasmus liliputanus].

Epidemiological survey and experimental studies on the mode of human infection with Echinochasmus liliputanus were carried out in Chenqiaozhou Village, Hexian County, Anhui Province in 1992. The results showed that humans could be infected through drinking unboiled water containing E.liliputanus cercaria or eating raw fish containing the metacercaria. The infection rates of E. liliputanus were 1.5% in the inhabitants who did not drink raw water, and 20.1% in those who used to drink raw water. It was only the cercaria, not the metacercaria that was detected in the water from ponds of this village. None of the investigated inhabitants ate uncooked fish. The results indicated that drinking raw water containing E.liliputanus cercaria was a major way through which a humans were infected and eating food containing the metacercaria was a minor mode of human infection.

Independent actions of asparagine and high levels of free Ca2+ in the induction of ornithine decarboxylase.

During growth stimulation of cells, Ca2+ and amino acids of the A, ASC and N transport systems are important for the induction of ornithine decarboxylase (ODC, L-ornithine carboxylase, EC 4.1.1.17). In order to clarify the relationship between Ca2+ and amino acids, we studied the induction of ODC by asparagine under three different Ca2+ states in H-35 rat hepatoma cells. First, in normal cells, extracellular Ca2+ above 0.1 mM and 10 mM asparagine separately stimulated ODC activity and their effects were approximately additive. In these normal cells, asparagine could act in the absence of medium Ca2+. TMB-8, a sequestered-Ca2+ release antagonist, had no effect on ODC induction whilst the asparagine action is sensitive to treatment with W7, a Ca-calmodulin antagonist, or lanthanum, a Ca2+ antagonist. Secondly, in cells treated with 0.5 mM EGTA in Ca(2+)-free medium, the asparagine action on ODC induction was blocked but the inhibition could be reversed by the addition of Ca2+ to the medium. Thirdly, ionomycin treatment in the absence of medium Ca2+ did not block the asparagine effect. Furthermore, in ionomycin-treated cells, the presence of high levels of medium Ca2+ increased ODC activity, but this increase was additive to, and could not replace, the action of asparagine. Our results indicate that the asparagine action does not depend on an increase of intracellular free-Ca2+.

Preparation of high purity IgG and its application in screening for clones positive for Vibrio succinogenes L-asparaginase gene.

A previous paper reported application of L-asparaginase IgG from Vibrio succinogenes in screening for positive clones. This paper describes the detailed procedure for preparation of high-purity IgG, an essential step of which involves coupling of a cell-free extract of host E. coli Y 1090 to Sepharose 4B beads and adsorption of the non-specific antigen components by passage of the polyvalent IgG through this affinity column. The IgG thus obtained exhibited improved specificity and was utilized as a radioimmunologic and horseradish peroxidase-linked immunologic probe in screening for positive clones. These IgG probes had the advantages of low background, high sensitivity, and good reliability.