Functional inactivation of the WTX gene is not a frequent event in Wilms' tumors.

For many years the precise genetic etiology of the majority of Wilms' tumors has remained unexplained. Recently, the WTX gene, mapped to chromosome Xq11.1, has been reported to be lost or mutated in approximately one-third of Wilms' tumors. Moreover, in female cases, the somatically inactivated alleles were found to invariantly derive from the active chromosome X. Consequently, WTX has been proposed as a 'one-hit' tumor suppressor gene. To provide further insights on the contribution of WTX to the development of the disease, we have examined 102 Wilms' tumors, obtained from 43 male and 57 female patients. Quantitative PCR analyses detected WTX deletions in 5 of 45 (11%) tumors from males, whereas loss of heterozygosity at WTX-linked microsatellites was observed in 9 tumors from 50 informative females (19%). However, in the latter group, using a combination of HUMARA assay and bisulfite-modified DNA sequencing, we found that the deletion affected the active chromosome X only in two cases (4%). Sequence analyses detected an inactivating somatic mutation of WTX in a single tumor, in which a strongly reduced expression of the mutant allele respect to the wild-type allele was observed, a finding not consistent with its localization on the active chromosome X. Overall, a functional somatic nullizygosity of the WTX gene was ascertained only in seven of the Wilms' tumors included in the study (approximately 7%). Our findings indicate that previously reported estimates on the proportion of Wilms' tumors due to WTX alterations should be reconsidered.

[Cesarean section, techniques and skin suture materials]. / Taglio cesareo, tecniche e materiali di sutura della cute.

AIM: The aim of the study was to compare the outcome, complications and costs of three skin suture techniques after Caesarean section. METHODS: The study sample was 310 women who underwent caesarean section between 2003 and 2007. The sample was divided into three groups: an intradermal suture with non-reabsorbable thread was applied in 98; metallic clips were placed in 90; 2-octyl-cyanoacrylate (2-OCA) glue was used for wound closure in 112. The sutures were checked at 4 days and 2 months after the operation to determine cosmetic outcome, patient compliance, strength of incision closure, allergic reactions, suture infection, and total cost of each technique. RESULTS: No substantial differences in strength of incision closure or cosmetic outcome between the techniques were found. Compliance was better in the group that received 2-OCA, while the total cost of suture alone was lower in the group that received the non-reabsorbable intradermal suture. CONCLUSION: The results suggests that following Caesarean section according to Stark, skin suture with 2-OCA glue has the advantage of greater patient compliance, while intradermal thread suture is less costly with a cosmetic outcome comparable to that of the other two techniques.

Refinement within single yeast artificial chromosome clones of a minimal region commonly deleted on the short arm of chromosome 7 in Wilms tumours.

Cytogenetic and molecular data indicate an involvement of genes mapped to the proximal portion of the short arm of chromosome 7 (7p) in Wilms tumours (WTs). We have analysed 38 WTs using a panel of eight microsatellite markers mapped to proximal 7p. Loss of heterozygosity (LOH) in tumour, compared with matched constitutional DNA, was identified in eight cases. To define better the minimal region commonly deleted in these tumours, they were analysed with nine additional markers, mapped within the region of interest. One tumour (case 30) showed LOH for only one marker (D7S510), while maintaining heterozygosity for the two immediately flanking loci (D7S555 and D7S668). This result was confirmed by fluorescence in situ hybridisation analysis, which showed that in the majority (65%) of nuclei from tumour 30 hybridising with a bacterial artificial chromosome clone containing the D7S510 locus, only one signal was visible. Noticeably, both markers defining the limits of the observed deleted region are simultaneously present within two distinct overlapping yeast artificial chromosome (YAC) clones mapped to chromosome bands 7p13-p14. This suggests that the maximum length of the missing DNA fragment was approximately 1.3 Mb, corresponding to the length of the smaller of the two YAC clones. In all other cases that showed LOH, the deletion encompassed the 7p13-p14 region. For this reason, we speculate that the identified interval contains a gene whose inactivation is important for the development of at least a fraction of WTs.

Cytogenetic and molecular characterization of T-cell acute lymphoblastic leukemia as a second tumor after anaplastic large-cell lymphoma in a boy.

We report a case of acute T-cell lymphoblastic leukemia which developed in a boy 8.5 years after successful treatment for anaplastic large-cell lymphoma. Cytogenetic and molecular characterizations of the second tumor were performed. The cytogenetic investigation revealed a complex pattern of karyotypic alterations, including double minutes, ring chromosomes, and a duplication of the p21-32 region of chromosome 1. The microsatellite DNA analysis excluded rearrangement or deletion of the TAL1 gene in the tumor cells; rearrangements of the MLL gene were excluded by Southern blot analysis. To the best of our knowledge, this is the first report of T-cell lymphoblastic leukemia arising after treatment of CD 30+ anaplastic large-cell lymphoma. The different T-cell receptor rearrangement evidenced in the two tumors indicates that this second malignancy most likely emerged de novo, but was plausibly related to the previous radiation and chemotherapy.

Molecular analysis of 1p32 genetic involvement in pediatric T-cell non-Hodgkin's lymphoma.

BACKGROUND AND OBJECTIVE: T-cell acute lymphoblastic leukemia (T-ALL) and lymphoblastic T-cell non-Hodgkin's lymphoma (T-NHL) are closely related disorders, and distinguishing between the two may be difficult. Cytogenetic investigations of large NHL series reported different recurring chromosomal alterations. Among these, aberrations of chromosome 1p seem to be associated with T-cell differentiation, the region most frequently involved in breakpoints being band 1p32-36. Deletions and translocations involving the same chromosomal region are frequently observed in T-ALL, in which one of the most common genetic changes is the breakage of the TAL1 gene, mapped to the 1p32 chromosomal region. The objective of this study was to assess the possibility of TAL1 involvement also in T-NHL. DESIGN AND METHODS: A series of 17 pediatric T-NHL patients was molecularly characterized by microsatellite markers analysis and by TAL1 gene microdeletions. RESULTS: TAL1 gene rearrangement was found in one case, while loss of heterozygosity (LOH) and microsatellite instability (MI) was identified in another case. INTERPRETATION AND CONCLUSIONS: Overall our findings indicate that, differently from T-ALL, neither TAL1 gene involvement nor LOH or MI at 1p32 appear particularly relevant in the oncogenic process of T-NHL transformation.

Retrospective analysis of ploidy in primary osseous and extraosseous Ewing family tumors in children.

AIMS: To retrospectively study the DNA content in a series of childhood Ewing Family Tumors (EFT), and to investigate its prognostic value. METHODS: The study was performed on a series of 27 EFTs (osseous Ewing's sarcoma, 18 cases; extraosseous Ewing's sarcoma, 2; peripheral neuroepithelioma, 4; Askin Rosai tumors, 3). Ploidy was investigated using both flow cytometry (FCM) and image cytometry (ICM) on tumor cell suspensions from formalin-fixed paraffin-embedded specimens or fresh frozen tissue obtained from the primary tumor at diagnosis. RESULTS: Ploidy was evaluable by FCM in all cases, and by ICM in 23/27. When fresh frozen tissue and paraffin-embedded samples from the same tumor were available for analysis, they yielded equal results. The rate of agreement between FCM and ICM was 82%. The majority of cases were diploid, and in the present series aneuploidy seemed to be associated with a poor outcome. CONCLUSIONS: These results suggest that aneuploidy could be an indicator of a bad prognosis in EFT; however, the small number of cases precludes any conclusion of statistical value. Larger retrospective studies on ploidy using archival material could be performed and their reliability is supported by the concordance of results from fresh and formalin-fixed tissue.

No evidence of WT1 involvement in a Burkitt's lymphoma in a patient with Denys-Drash syndrome.

BACKGROUND: We previously reported the case of a patient affected with Denys-Drash syndrome (DDS), who developed disseminated EBV-related Burkitt's lymphoma (BL) after kidney transplantation. Here, we describe the molecular characterisation of the WT1 gene in the constitutional and tumour DNA of this patient. PATIENTS AND METHODS: WT1 exons 2 to 10 were sequenced in constitutional and tumour DNAs. By Southern blotting the latter was also investigated for the presence of gene rearrangements. Gene expression analysis in tumour cells was performed by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: A germline missense mutation affecting one of the zinc finger domains of the gene, and previously reported in other DDS cases, was observed. No alterations of the constitutionally wild-type WT1 allele and no expression of the gene were observed in BL cells. A small group of BLs from other paediatric patients showed a variable expression of WT1. CONCLUSIONS: Our findings indicate that WT1 is unlikely to be involved in the onset of BL in our case. However, a possible role of the gene in at least a subset of these lymphoproliferative diseases may be suggested.

Disseminated Burkitt's lymphoma after kidney transplantation: a case report in a boy with Drash syndrome.

PURPOSE: We discuss the clinical, laboratory findings and treatment of a boy who developed Burkitt's lymphoma (BL) after renal transplant and some issues about lymphoproliferative disorders after transplantation. METHODS: A 6-year-old boy with Drash syndrome developed disseminated Burkitt's lymphoma 38 months after transplantation for renal failure. Immunosuppressive therapy had consisted of prednisolone and cyclosporine. Polychemotherapy was initiated. RESULTS: Polychemotherapy induced rapid and complete remission of the disease without major side effects despite the renal transplant allograft and prolonged immunosuppression. CONCLUSIONS: A child with posttransplantation B-cell high-grade lymphoma can be successfully treated with the same chemotherapy regimen used for ordinary cases.

Lipoprotein(a) and other lipids after oophorectomy and estrogen replacement therapy.

OBJECTIVE: To assess the effect of surgical menopause and subsequent estrogen replacement therapy (ERT) on lipoprotein(a) [Lp(a)] and common lipids. METHODS: In 24 healthy premenopausal women, lipids (total cholesterol, low-density lipoprotein [LDL] and high-density lipoprotein [HDL] cholesterol, and triglycerides) and gonadotropins (FSH, LH) were measured the day before hysterectomy with bilateral oophorectomy and then after 1, 2, and 3 months. Blood was also drawn after 2, 4, 6, and 15 days to exclude the effect of surgery on Lp(a). In 19 women who volunteered for ERT, the lipid profile was assayed again after 3, 6, and 12 months of treatment. RESULTS: Lipoprotein(a) levels rose significantly over the 3 months after surgery, from a mean +/- standard deviation (SD) 5.7 +/- 6.1 mg/dL to 10.4 +/- 9.2 mg/dL. Total cholesterol and LDL cholesterol levels increased significantly over the first 2 months; HDL cholesterol decreased significantly during the 3 months of follow-up (by 10, 17, and 20%) (P < .001). Plasma triglycerides did not change after surgery. Three months following ERT, Lp(a) and total cholesterol were significantly decreased (28 and 11%, respectively), as was LDL cholesterol (33%) after 6 months. High-density lipoprotein cholesterol increased by 24% after 6 months of treatment, and triglycerides rose significantly in the year of therapy (37%). CONCLUSION: These findings suggest that surgical menopause induces atherogenic changes in the lipid profile in 3 months and that ERT soon reverses them.

Mapping of a putative tumor suppressor locus to proximal 7p in Wilms tumors.

Different findings suggest that alterations of chromosome 7 genes play a role in the development of Wilms tumors. To define the positions of these genes, we have accomplished a combined cytogenetic and molecular study on 11 sporadic Wilms tumors. In one case, where both chromosomes 7 were rearranged, the karyotypic picture was consistent with the presence of a tumor suppressor gene at 7p15. To test this hypothesis, a loss of heterozygosity analysis was performed using microsatellite markers. This revealed a common region of allele losses mapped to the proximal short arm of chromosome 7 and defined the position of the gene(s) involved in Wilms tumors within an interval of approximately 25 cM.

Comparison of postoperative complications after Küstner and Pfannenstiel transverse suprapubic incisions.

We compared postoperative morbidity with the Küstner (n = 53) and Pfannenstiel (n = 131) incisions in a consecutive series of women undergoing surgery for benign gynecological conditions. The incidence of febrile morbidity (15.1% vs 23.7%, chi 2(1) = 1.19, P = 0.28) and wound infection (5.7% vs 9.2%, P = 0.56, Fisher's exact test) were higher in the Pfannenstiel then in the Küstner group, but neither difference was statistically significant. One suprafascial hematoma was observed after a Küstner incison compared with eight subfascial hematomas after a Pfannenstiel incision (1.9% vs 6.1%, P = 0.45, Fisher's exact test). The postoperative hospital stay was statistically significantly lower in the Küstner than in the Pfannenstiel group (6.3 +/- 1.4 vs 7.1 +/- 1.2 days, P < 0.01, Student's t test). The Küstner incision warrants further evaluation and usage.

Allelotyping in Wilms tumors identifies a putative third tumor suppressor gene on chromosome 11.

An analysis of loss of heterozygosity for markers on both the short and the long arm of chromosome 11 was performed in 24 sporadic Wilms tumors. Six cases (25%) showed allelic losses involving the entire chromosome. In one case (4%) the loss was restricted solely to the WT1 gene on band p13. Two cases (8%) displayed allelic losses for WT1 and for markers on band p15.5, where the putative tumor suppressor gene WT2 has been mapped, but retained heterozygosity for markers on the long arm. In three tumors (13%) the loss of heterozygosity involved markers mapped to chromosomal regions p15.5 and q23.3-qter, but did not affect WT1 and markers on q12-q13. Altogether, the proportion of cases showing allelic losses at the distal region of 11q (37%) was comparable to that of cases with LOH affecting the WT1 (37%) or the WT2 (46%) loci, thus suggesting the existence of a third chromosome 11 tumor suppressor gene involved in the pathogenesis of Wilms tumors.

[Behavior of Candida albicans during a period of nutritional deprivation. Preliminary results]. / Comportamento di Candida albicans durante un periodo di carenza nutrizionale. Risultati preliminari.

Cells of Candida albicans, after 24 hours of growth in YM, were starved, alternatively, in citrate buffer, physiological solution, MMS deprived of glucose or ammonium sulphate. The eventual growth was monitored by determining the absorbance at 675 nm. Simultaneously, the cell morphology was also controlled. In a second series of experiments, the C. albicans cells taken from YM were starved for 72 hours in one of the mediums as stated above, and then reinoculated in MMS liquid without, alternatively, glucose or ammonium sulphate. Again the eventual growth was monitored as in the above method. The achieved results indicate the presence of a reserve of nitrogen, which can be utilized when a source of C is given to the cell. We therefore discuss the apparent lack of glucidic reserve and we propose a method for the consumption of nitrogen reserve. The aim of the work is to define how to obtain cells that contain the smallest amount possible of endogenous reserve.