Spinal deformity after intra-operative radiotherapy for paediatric patients.

The purpose of this study was to clarify the incidence and characteristics of late-onset complications of the spine in children who underwent intra-operative radiation therapy (IORT) for common paediatric malignant tumours. 12 children with more than 4 years of follow-up after IORT were included and, in 11 of these, thoracic and/or lumbar spines were irradiated. To compare doses of irradiation to the spine with the resulting deformities, dose simulations of IORT were carried out on two selected cases using a radiation treatment planning system with a pencil-beam algorithm. The mean follow-up period was 135 months (range, 53-234 months). Radiographic reviews found spinal deformity in six patients. Only one patient was symptomatic and the spinal deformity was severe (Grade 3), whereas spinal deformity was mild in the remaining five patients without clinical symptoms (Grade 1). In all of the six patients, anterior wedge-shaped deformity was dominant, and scoliosis was found in only two patients. In one particular case with nephrectomy, irradiation had penetrated much deeper than usual at the site of nephrectomy, and dose distribution was asymmetric, causing clinically significant spinal deformity with scoliosis. In conclusion, specific deformities of the spine observed after IORT can be explained on the basis of dose distribution of the electron beam to the spine.

Light-induced pigment aggregation in xanthophores of the medaka, Oryzias latipes.

The response mechanism of medaka xanthophores to light was examined at the cellular level. Innervated and denervated xanthophores of adult medakas responded to light (9,000 lux) within 30 sec by pigment aggregation, and this aggregation was not mediated through alpha-adrenoceptors on the cell membrane. Maximum sensitivity to light was at wavelengths of 410-420 nm, and the direct effect of light was reversible. Xanthophore responsiveness to light in summer was higher than that in winter. Ca2+ and calmodulin were not involved in the response, but rather, an important role for cAMP and phosphodiesterase (PDE) was suggested. It seems likely that photoreception by visual pigment which is sensitive to light at wavelengths of 410-420 nm increases PDE activity, probably via a G-protein, such as occurs with visual cells in the retina, which causes a decrease in levels of cytosolic cAMP, in turn leading to pigment aggregation within medaka xanthophores.

Heterotopic pancreas as a rare cause of gastrointestinal hemorrhage in the newborn: report of a case.

We report herein a female infant in whom a heterotopic pancreas in the stomach caused gastrointestinal hemorrhage during the newborn period. Endoscopy was essential for ruling out other causes of hemorrhage and to follow the patient until the time of elective surgery at 6 months of age. Heterotopic pancreas should be borne in mind as a rare cause of gastrointestinal hemorrhage in the newborn when an unexplained gastrointestinal hemorrhage continues, and the lesion was discussed in relation to our experience of upper gastrointestinal endoscopies and a review of the literature.

[Endocrinological and metabolic disorders in the patients with massive bowel resection in the newborn period].

Massive (70%) resection of the small bowel was performed in seven newborn infants. Follow-up study was undertaken in four of these patients and the growth and endocrinological status were evaluated. Body weight and height of three patients were below 50th percentile. On glucose tolerance test, peak IRI was low and insulinogenic indices were below normal range. These findings suggest that there are some mechanisms which suppress the secretion of insulin. Pituitary and thyroid function were within normal limits but secondary sexual manifestation was not seen in a fifteen year old boy. Hyperoxaluria was seen in three children, and one had kidney stone with some impairment of renal function.

[Late complications of Hirschsprung's disease and anorectal manometry].

In 84 patients treated with Soave-Denda method, informations of their bowel habits were obtained through a questionnaire. These patients were classified into 4 groups (A; less than 3 years, B; 3 to 6 years, C; 7 to 9 years and D; more than 10 years after the operation) and analyzed by comparing their bowel habits with those of normal children in corresponding age group. Some late complications were demonstrated although 80% of patients followed for more than 10 years had a normal bowel habit. Frequent incontinence (more than once a week) was found in 18.2% of group B and 5% of group C. No patients has incontinence in group D. Frequent soiling (more than once a week) was demonstrated in 58.8% of group B, 30% of group C and 23.1% of group D although the incidence of frequent soiling ranged from 0.5 to 2.1% in normal children of these age group. Manometry to detect a rectonal reflex was useful to evaluate anorectal function because patients who showed normal reflex had excellent bowel habits without incontinence. These unexpected high incidence of late complications indicate that postoperative training of defecation is extremely important to give normal bowel habit to the patients with Hirschsprung's disease.

The effect of fibronectin on cholinergic differentiation of the fetal colon.

In an effort to determine the factors which affect cholinergic development after completion of migration of neural crest cells to the colon, the extracellular matrix constituents, fibronectin, collagen IV, laminin, and heparan sulfate were studied during this later postmigration stage of differentiation. Distal colons of the 14 1/2 day embryo rat were incubated in vitro with antibodies to the above constituents or with fibronectin alone. Cholinergic function of the colon was assessed by acetylcholinesterase staining and choline acetyltransferase activity. When 100 micrograms/mL of fibronectin was added to the medium, the choline acetyltransferase activity was enhanced; when antibody to fibronectin (50 or 100 micrograms/mL) was added, acetylcholinesterase staining and choline acetyltransferase activity were inhibited. Addition of anti-laminin, anti-collagen IV, or anti-heparan sulfate did not affect either acetylcholinesterase staining or choline acetyltransferase activity. Fibronectin may be an important factor in cholinergic differentiation of the enteric nervous system during the postmigration stage of development.

Subrenal capsule assay to test the viability of parenterally delivered müllerian inhibiting substance.

Production of bovine müllerian inhibiting substance (MIS) has been increased to allow generation of large quantities of biologically active purified material. The limited MIS previously available allowed only pretreatment of tumors prior to colony inhibition or implanting in nude mice. In preparation for posttransplantation tumor treatment, a subrenal capsule assay, which was first used against human tumors heterotransplanted into nude mice and subsequently against those heterotransplanted into immunocompetent mice, was adapted to determine (1) if MIS preparations could traverse the bloodstream without degradation and (2) the optimal dose required to produce a biologic effect. Urogenital ridges from female 14-day-old rat embryos were transferred atraumatically to small pouches beneath the renal capsule of the immunocompetent male CDF1 mice. The cranial-caudal orientation of the ridge with its müllerian duct was maintained. Over the next 72 hours, the mice were injected via the tail vein with 0.1 mL of an MIS-containing solution over a 100-fold concentration range. After three days, the kidneys were removed and shaved just below the ridge, which was then placed in soft agar for orientation and subsequent serial sectioning. After fixation, dehydration, and paraffin embedding, sections were stained and regression of the müllerian duct was graded and compared according to concentration and number of MIS doses administered. Regression diminished from almost complete (4+) at the highest dose, to minimal (1 to 2+) at 1/100 of that dose. Heat-inactivated and vehicle controls caused no regression of the müllerian ducts.(ABSTRACT TRUNCATED AT 250 WORDS)

Phosphorylation events during Müllerian duct regression.

Regression of the fetal rat Müllerian duct in vitro was stimulated by sodium fluoride in the absence of Müllerian inhibiting substance. The action of Müllerian inhibiting substance was inhibited by sodium vanadate, adenosine 5'-triphosphate, and several related nucleotides in the presence of manganese ions. Epidermal growth factor specifically inhibited the substance, but only with manganese ions present. Insulin, platelet-derived growth factor, and nerve growth factor had no effect. These results suggest that dephosphorylation of membrane proteins mediates the action of Müllerian inhibiting substance.

Production of monoclonal antibodies for affinity purification of bovine mullerian inhibiting substance activity.

Two monoclonal antibodies (IG8 and IG10) specific for Mullerian inhibiting substance (MIS) were obtained from the fusion between myeloma cell line SP2/0 and spleen cells from an A/J mouse immunized with partially purified MIS. The resulting hybridomas were screened by a solid-phase RIA and two lines were selected and cloned. Both MAbs IG8 and IG10 subsequently demonstrated specificity for MIS by their ability to inhibit biologically active MIS by precipitation with a second antibody, directly block MIS activity in the organ culture assay, and adsorb and elute active MIS when coupled to a solid support. SDS-polyacrylamide gel electrophoresis of affinity purified MIS demonstrated a major band at 140 kD in unreduced gels and two bands with approximate molecular weights of 70 and 74 KD following reduction. Protein bands were localized either directly by silver staining or on immunoblots developed with radiolabeled anti-MIS MA.

Extended dissection of the portahepatis and creation of an intussuscepted ileocolic conduit for biliary atresia.

This paper introduces a new operation for biliary atresia that establishes successful biliary drainage by extending the portahepatic dissection, and decreases ascending cholangitis by incorporating a nonrefluxing intussuscepted draining conduit. An analysis of the postoperative results is presented. The usual dissection of the portahepatis is extended to include the area between and beneath the branches of the right portal vein to incorporate all potentially usable remnants of the intrahepatic ducts. Ascending cholangitis is decreased by interposing an intussuscepted ileocolic segment between the portahepatis and the abdominal wall. This extended dissection of the portahepatis was performed since 1978 in 15 infants with noncorrectable biliary atresia, and bile drainage was achieved in all. In ten infants an intussuscepted ileocolic conduit was constructed. Ascending cholangitis in these ten infants has been either completely absent or easily controlled by antibiotics. The draining bile was highly concentrated due to the water-absorbing capacity of the interposed colonic segment; therefore, fluid and electrolyte disturbances, which develop frequently in patients having jejunal conduits, have never been encountered.

Mullerian inhibiting substance fractionation by dye affinity chromatography.

Mullerian inhibiting substance (MIS), a large glycoprotein secreted by the fetal and neonatal testis, is responsible for regression of the Mullerian ducts in the male embryo. This fetal growth regulator has been purified more than 2000-fold from crude testicular incubation medium following fractionation on a triazinyl dye affinity support. A high yield of 60% recovered activity was achieved in the absence of exogenous carrier protein by stabilizing MIS with 2-mercaptoethanol, EDTA, and Nonidet-P40 and eliminating losses in the handling and concentration of MIS fractions. Although affinity elution with nucleotides has proved successful in other systems, MIS could not be eluted with ATP, GTP, or AMP, with or without divalent metal ions. Nucleotide elution, however, does remove contaminating proteins prior to MIS recovery with high ionic strength. The 2000-fold-purified MIS fraction, although not homogeneous, shows a reduction-sensitive band after SDS-gel electrophoresis that has been proposed to be the MIS dimer.