Shared Copy Number Variation in Simultaneous Nephroblastoma and Neuroblastoma due to Fanconi Anemia.

Concurrent emergence of nephroblastoma (Wilms Tumor; WT) and neuroblastoma (NB) is rare and mostly observed in patients with severe subtypes of Fanconi anemia (FA) with or without VACTER-L association (VL). We investigated the hypothesis that early consequences of genomic instability result in shared regions with copy number variation in different precursor cells that originate distinct embryonal tumors. We observed a newborn girl with FA and VL (aplasia of the thumbs, cloacal atresia (urogenital sinus), tethered cord at L3/L4, muscular ventricular septum defect, and horseshoe-kidney with a single ureter) who simultaneously acquired an epithelial-type WT in the left portion of the kidney and a poorly differentiated adrenal NB in infancy. A novel homozygous germline frameshift mutation in PALB2 (c.1676_c1677delAAinsG) leading to protein truncation (pGln526ArgfsX1) inherited from consanguineous parents formed the genetic basis of FA-N. Spontaneous and induced chromosomal instability was detected in the majority of cells analyzed from peripheral lymphocytes, bone marrow, and cultured fibroblasts. Bone marrow cells also showed complex chromosome rearrangements consistent with the myelodysplastic syndrome at 11 months of age. Array-comparative genomic hybridization analyses of both WT and NB showed shared gains or amplifications within the chromosomal regions 11p15.5 and 17q21.31-q25.3, including genes that are reportedly implicated in tumor development such as IGF2, H19, WT2, BIRC5, and HRAS.

The role of RET genomic variants in infantile hypertrophic pyloric stenosis.

Infantile hypertrophic pyloric stenosis (IHPS) is a common childhood pathology affecting approximately 1-5 children pro 1000 newborns, with a genetic background as suggested by the familial occurrence. RET is a candidate gene for IHPS due to its role in the development of the intrinsic innervation and ganglia of the smooth musculature and the association of RET variants with another motility disorder (Hirschsprung's disease). Accordingly, we investigated RET-IHPS associations through sequencing of the complete RET coding region in 32 IHPS patients. Genotype frequencies were compared between patients and 48 controls using the Cochran-Armitage trend test or Fischer's test for exact p-values. We found 19 RET variants in IHPS, including polymorphisms in the promoter region (c.-200 G>A and c.-196 C>A). There was no statistically significant difference between the frequencies of the variants in both groups. There was no deviation from the Hardy-Weinberg equilibrium, yet a significant correlation (linkage disequilibrium) for variants in the promoter region, in exons 11, 13, 14 and 19 and in the 3' UTR. We conclude that RET variants are present in IHPS patients yet show no significant statistical association with the IHPS phenotype, suggesting at best an adjuvant role for RET in IHPS.

[Diagnosis of appendicitis using harmonic ultrasound imaging]. / Sonografische Diagnostik der Appendizitis bei Kindern - Verbesserte Darstellung mittels Harmonic Imaging.

INTRODUCTION: Harmonic Imaging (HI) is a new Ultrasound (US) modality with high contrast and spatial resolution. Aim of this study was to compare fundamental imaging (FI) and HI in the diagnosis of appendicitis. PATIENTS AND METHODS: In 50 children (male : female, 25 : 25; mean age: 9.9 years) prior to an appendectomy US of the right lower quadrant was performed in both FI and HI (Tissue Harmonic Imaging - THI - Sonoline Elegra, Siemens, 7.5 MHz linear). The images were compared with respect to delineation of surrounding, contour, wall and content of the appendix. RESULTS: In 43 / 50 (86 %) patients diagnosis of appendicitis was confirmed histologically (sensitivity 98 %, specificity 94 %). In 37 / 43 patients the appendix was depicted using FI. With HI this number was 40 / 43. HI was significantly better in delineating the contour, wall, mucosa and content of the appendix (p < 0.01). HI also exceeded in demonstrating free fluid, mesenterial lymph nodes and surrounding echogenicity. CONCLUSION: HI of appendicitis provides images with significantly better quality than FI. When both imaging options are available harmonic imaging modality should be the preferred choice for scanning the appendix.

Transient pseudohypoaldosteronism with complex malformation of internal genitalia. A case report.

At the age of 3 weeks, a girl presenting with acute dehydration was admitted to our hospital. Clinical and laboratory findings revealed malformations of the genitourinary tract, an acute urinary infection and electrolyte disturbances (severe hyponatremia at 115 mmol/l and mild hyperkalemia at 5.6 mmol/l). According to anamnestic data, the child was born to healthy, nonconsanguineous parents. Vaccum extraction was done in the 38th gestational week due to pathological cardiotocography (CTG) findings. Auxological parameters were within the normal range for gestational age. Normal values for 17-OH progesterone and ACTH ruled out congenital adrenal hyperplasia (CAH). Pathologically high aldosterone and plasma renin activity (PRA) confirmed the diagnosis of pseudohypoaldosteronism with salt-wasting crisis which proved to be transient.

Synthesis of beta-difluorine-containing amino acids.

A convenient strategy was developed to prepare several beta-difluoroamino acids. As exemplified by the synthesis of 3,3-difluoro-L-homocysteine, 3,3-difluoro-L-homoserine and 3,3-difluoro-L-methionine, the reaction sequence all started from L-isoascorbic acid. This approach holds potential to be extended to make other beta-difluorine-containing amino acids.

Affinity labeling of Escherichia coli glucosamine-6-phosphate synthase with a fructose 6-phosphate analog--evidence for proximity between the N-terminal cysteine and the fructose-6-phosphate-binding site.

Glucosamine-6-phosphate synthase (GlcNP-synthase) catalyzes the formation of glucosamine 6-phosphate from fructose 6-phosphate using the gamma-amide functionality of glutamine as the nitrogen source. In the absence of glutamine, GlcNP-synthase was recently found to catalyze the formation of glucose 6-phosphate corresponding to a phosphoglucoisomerase-like activity. Here we report active-site directed, irreversible inhibition of Escherichia coli GlcNP-synthase (k(inact) = 0.60 +/- 0.05 min(-1), Kirr = 1.40 +/- 0.20 mM) by anhydro-1,2-hexitol 6-phosphates previously known as irreversible inhibitors of phosphoglucoisomerase. Enzyme inactivation with the tritiated affinity label, followed by tryptic digestion and purification of the radioactive fragments, allowed identification of three peptides. Two of them, accounting for 54% of the recovered radioactivity, are believed to result from the nucleophilic attack of side-chain carboxylates of Glu255 and Glu258 and thiol of Cys300 of the fructose-6-phosphate-binding site on the epoxide functionality of the inhibitor. The major peptide corresponds to derivatization of the N-terminal cysteine from the glutamine-binding site by the inhibitor. These results provide evidence for the close proximity of glutamine and fructose-6-phosphate-binding sites recently suggested by Bearne [Bearne, S. L. (1996) J. Biol. Chem. 271, 3052-3057].

Value of a score in the electromanometrical diagnosis of neuronal intestinal dysplasia.

Anorectal manometry was performed in a series of 8 children where NID was histochemically confirmed or suspected. The results were transferred to a electromanometric score theoretically worked out previously. Clinical symptoms as well as histological and manometric findings have been compared concerning the meaningfulness of the electromanometric score in the diagnosis of NID. It was impossible to make a statistical statement due to the small number of patients we examined. It seems, however, that certain electromanometric parameters in NID appear regularly which, when combined in a score, make manometric diagnosis of NID sufficiently reliable.

Day case surgery in childhood from the parents' point of view.

294 parents whose children underwent day case surgery received a questionnaire covering the topics postoperative analgesia, memory of pain, sleep patterns and assessment of the time spent in the hospital. A total of 80.6% of the questionnaires were returned. We were not able to recognize any indications of behavioral disturbances, in particular disturbances of sleep. The need for analgesics was surprisingly low following release from the hospital. However, even on the third postoperative day 16.8% of the children still had unpleasant or strong memories of the pain they had experienced. Improvement in this area might be achieved by the application of nerve blocks, possibly in combination with analgesics. Further possibilities for improvement exist with premedication. The long waiting periods between hospital admittance and commencement of surgery were found to be very unpleasant. During surgery psychological counseling of the parents should be made possible.

[Adrenocortical nodular hyperplasia as a cause of Cushing syndrome in the neonatal period]. / Adreno-corticale noduläre Hyperplasie als Ursache eines Cushing-Syndroms in der Neugeborenenperiode.

The case report of a female child born preterm (30th wk of gestation) who developed symptoms of Cushing's syndrome beginning in the neonatal phase is presented. The disease was caused by a unilateral adreno-cortical nodular hyperplasia and was successfully treated by unilateral adrenalectomy. Preoperative treatment with ketoconazole and metyrapone proved to be effective. Symptoms of Cushing's disease including hypertrophic cardiomyopathy were completely reversible within one month after surgery.

Studies to be conducted before projected operations from the pediatric surgeon's point of view.

To rationally discuss the "need for preoperative laboratory investigations prior to performing an operation that can be pre-planned", two questions must be considered: 1. What kind of complications were encountered during or after 25,872 anesthesias performed in a 20-year period? 2. What kind of diseases were recognized only postoperatively which apparently could not be diagnosed preoperatively via case history and clinical examination? The results of these analyses are tabulated and assessed. It could be shown that the case history and the clinical examination of the patient on the day of surgery rank before all laboratory tests. With regard to peroperative respiratory disturbances it could be proven that children within the first trimester are particularly susceptible to such alterations. Furthermore, it could be demonstrated that within this group premature babies are more susceptible to this kind of complication than babies born at date. With regard to the incidence of preoperative respiratory disturbances in children with or without infection of the upper respiratory tract, it was shown that children without such symptoms who had however undergone an infection three to four weeks before the operation, were more susceptible to peroperative respiratory disturbances than children without any infection or even children suffering from an acute infection. Taking the physiological Hb decrease at the end of the first trimester as an example, questions on the optimal date for surgery are discussed. Preoperative handling is discussed on the basis of the blood glucose level of 500 children after a fasting period of 3-4 hours.