Pediatric and Developmental Pathology

Congenital diaphragmatic hernia (CDH) in humans carries high mortality/morbidity attributed to associated pulmonary hypoplasia. An understanding of the effects of CDH on fetal lung growth is important for development of successful treatments. This study aimed to quantitate structural differences between normal and CDH-affected preterm lamb lungs. We hypothesized that (a) pulmonary hypoplasia is present in preterm CDH-affected lungs; (b) the relative degree of pulmonary hypoplasia increases with gestation; and (c) the left upper lobe (LUL) is affected most.

Fetal lambs were allocated to two groups. One group underwent surgery (72–74 days gestation) inducing CDH. Both groups (n = 7, n = 7) were delivered by cesarean section at 129 days (term: 145–149). Lungs were obtained at autopsy, were inflation-fixed, processed for histology, and morphometry was performed.

Preterm lungs of CDH-affected lambs in comparison to those of normal lambs demonstrated a reduction in the following: lung weight (37.7 g vs. 116.3 g); lung weight:body weight (0.012 vs. 0.040); fixed lung volume (33.6 ml vs. 96.9 ml); gas-exchange surface area (4.56 m² vs. 13.70 m²); parenchyma:nonparenchyma (59:41 vs. 72:28); and parenchymal airspace:tissue (16:84 vs. 35: 65). Non-parenchyma connective tissue was increased (58%), airspaces were more numerous (1077/mm²) and smaller (perimeter 76.6 μm), gas-exchange surface density (2394 cm⁻¹) was greater and capillary loading (0.04 ml/m²) was reduced compared to preterm normal lung (49%; 778/mm²; 108.7μm; 2003 cm⁻¹, 0.11 ml/m², respectively). The LUL was affected most.

These data quantitate pulmonary hypoplasia in preterm CDH-affected lambs. Comparisons with published data indicate increasing relative hypoplasia as gestation proceeds. Fetal interventions will affect lung development, depending on timing, with intervention still likely to be worthwhile during late gestation.

The Pró-Natal project is a collaborative initiative that aims to improve maternal and infant health in a deprived community in Natal, Northeast Brazil. To assess the perinatal and infant mortality in this population of 40,000, we have collected over a 2-year period a consecutive series of 39 autopsy examinations on deaths under 1 year of age. During this period there were 2212 live births in the study population. The 14 perinatal deaths are described using the Wrigglesworth classification, and the 25 infant deaths, using a clinicopathological system.

The contribution of normally formed stillbirths was small (14%), which probably reflects the underreporting of stillbirths in this community. The most common cause of death in the live births was complications of prematurity (43%). Specific causes (22%) of perinatal deaths were predominantly infections, including one case of congenital syphilis. Perinatal asphyxia was diagnosed in 14%, and there was one case (7%) of a chromosome abnormality. Infant deaths were predominantly due to respiratory (45%) and gastrointestinal infections (28%), with chronic malnutrition as an underlying cause in 80% of cases.

Prenatal care could theoretically have prevented three of the perinatal deaths, and a further six deaths could have been avoided by improved management of labor and the immediate neonatal period. Prevention of malnutrition and improved treatment of acute infections would contribute to a reduction in infant mortality in this population. The Pró-Natal project will use these data to design preventative interventions to reduce perinatal and infant mortality in this community.

The expression of the antibody Melan-A in 27 benign melanocytic skin lesions (10 congenital nevi, 10 Spitz nevi, and 7 pigmented spindle cell nevi) was compared to that of S100 protein and HMB-45. To evaluate the role of Melan-A in differentiating melanocytic and nonmelanocytic lesions we assessed a number of benign nonmelanocytic skin lesions including neurofibromas, granular cell tumors, and dermatofibromas.

Melan-A had an identical staining pattern to S100 protein in the melanocyte population of all lesions, but had the advantage of only staining cells of melanocytic lineage and no other cell types. HMB-45, although staining the junctional components of all lesions with a junctional component, showed varied intensity and distribution in the dermal components. Melan-A is much cleaner than S100 protein, having no background staining, and in skin appears to be specific for melanocytes. The nonmelanocytic lesions did not express Melan-A.

The aims of this study were to determine whether immunohistochemical staining for C9 can demonstrate myocardial necrosis in the fetus and neonate. Hearts from cases of stillbirth or neonatal death with confirmed myocardial necrosis (in neonates) or with ischemic lesions outside the heart (in neonates and stillborns) were stained immunohistochemically with antibodies to C9. All five cases with confirmed myocardial infarction showed positive immunohistochemical staining for C9, largely localized to the infarcted areas. The youngest subject was born at 24 weeks gestation and died at 4 days of age. One of two neonates without myocardial necrosis on H&E staining but with pathological evidence of ischemic lesions elsewhere showed staining of scattered fibers. Six out of ten hearts from macerated stillborn infants showed varying degrees of positive staining. Immunohistochemical staining for C9 detects myocardial necrosis in neonates of a gestational age of 24 weeks or more. C9 is also demonstrable immunohistochemically in macerated stillborns, and this is likely to represent myocardial necrosis. The method is of great potential value in the investigation of cardiac ischemia in the fetal and perinatal period.

The objective of this study was to test the hypothesis that macrosomic infants of nondiabetic mothers have beta-cell hyperplasia in their pancreases. Pancreatic tissues were examined from 10 macrosomic fetuses and liveborn infants and from 10 comparison cases matched for gestational age and gender. None of the mothers had a history of diabetes and all had normal glucose screening during pregnancy. Tissues were stained with hematoxylin and eosin and a monoclonal antibody against beta cells and were analyzed using an image analysis program to evaluate the size and surface area of beta-cell clusters. Brain/liver weight ratios were calculated and compared. The total surface area and cluster size of beta cells in the pancreases of macrosomic subjects were significantly larger than in the comparison pancreases. The study subjects lacked macroscopic and histopathologic findings expected in infants of diabetic mothers. We conclude that some macrosomic fetuses and infants of nondiabetic mothers manifest beta-cell hyperplasia. This corresponds to the higher insulin levels in macrosomic infants of nondiabetic mothers described in previous clinical studies. In macrosomic fetuses the stimulus for beta-cell hyperplasia may not involve aberrant maternal glucose levels.

The objective of this study was to determine if perioperative elevation of cardiac troponin I (cTnI) predicts mortality in infants and children after surgical correction of congenital heart defects. One hundred infants and children having open heart surgery were studied. Blood samples for cTnI analysis were collected before cardiopulmonary bypass (CPB) and at 4, 8, 12, and 24 h after initiation of CPB. Demographic information, cardiac defect, repair performed, duration of CPB, complications, and outcome were recorded. Cardiac defects were categorized as atrial septal defect (ASD), ventricular septal defect (VSD), hypoplastic left heart syndrome (HLHS), complex, and “other.” Baseline cTnI was significantly lower in survivors (mean 0.42 ng/ml, median 0.35 ng/ml) than in nonsurvivors (mean 1.89, median 1.30), p = 0.0001. Baseline cTnI was significantly higher in the HLHS group (mean 1.47, median 1.10) than in all other subgroups (mean 0.62, median 0.35), p ≤ 0.009. There were no significant differences between survivors and nonsurvivors at the remaining sampling times. Children who died from cardiac failure (n = 2) were more likely to have 4 h cTnI > 125 ng/ml compared to survivors (2 of 90). Within cardiac defect subgroups, 4 h cTnI was significantly higher in the complex group (mean = 53.51, median = 32.30) than in the ASD (mean = 23.84, median = 19.85) and other (mean = 21.59, median 21.50) subgroups. Perioperative measurement of cTnI identifies children within specific cardiac defect subgroups at risk of mortality after cardiac surgery. We speculate that detection of myocardial injury may decrease mortality and morbidity in children with complicated congenital cardiac lesions by leading to improvements in perioperative management.

We describe two prepubertal girls with oncogenic rickets. The first patient, 9 years of age, presented with recent-onset lower-extremity pain. The second girl, presented at 4 years of age following a 9-month period of muscle weakness, bone pain, and poor linear growth. Laboratory analyses in both patients revealed hypophosphatemia and hyperphosphaturia; elevated circulating alkaline phosphatase activity was present in one of them. Radiographic evidence of a generalized rachitic process was evident in both cases. Computerized tomography of the paranasal sinuses and facial bones in patient 1 revealed a small lesion eroding through the inner table of the left mandibular ramus. Microscopic examination of this mass revealed a spindle cell neoplasm with chondroid material, dystrophic calcification, and both osteoclast-like and fibroblast-like cells. Prominent vascularity and marked atypia were present. These features are consistent with a phosphaturic mesenchymal tumor of the mixed connective tissue variant. In the second patient, computerized tomography revealed a lytic lesion located in the right proximal tibia, with histologic features consistent with a phosphaturic mesenchymal tumor of the nonossifying fibroma-like variant. Resection of each tumor resulted in rapid correction of the phosphaturia and healing of the rachitic abnormalities. A careful search for small or occult tumors should be carried out in cases of acquired phosphaturic rickets.

We explored the mechanism of normal hepatic growth in children by evaluating the proliferation index (with Ki-67 antibody) and apoptosis index (with the ApopTag^™ kit). The proliferative index is almost constant, but the level of apoptosis is very low in childhood and increases with age. Thus the growth of the liver in children is not due to a high proliferation index, but to a very low apoptosis index.

Inborn metabolic errors causing lysosomal storage, such as β-galactosidase deficiency (G_m1 gangliosidosis [G_m1]), have well-recognized effects on cellular function and morphology. In some classically “neuronal” storage diseases, including G_M1, neuroradiologic observations of infants have suggested a delay in myelination on the basis of persistently “immature” signal intensities monitored over time. We sought to evaluate in a semiquantitative fashion the pattern and degree of myelination in two infantile G_m1 patients, one boy and one girl, autopsied at 15 months of age. We assigned myelination degrees for defined sites on an ordinal scale of 0 to 4, and compared them to published population-based values for autopsied infants. In both patients, earlier-myelinating structures were comparable in development to that expected for postconceptional age, whereas later-myelinating structures were delayed. These data correlate well with the neuroradiologic diagnosis of myelination delay in these infants and suggest that the metabolic defect has a primary influence on myelin development, in addition to effects related to neuronal storage. Furthermore, our analysis by light and electron microscopy and lectin histochemistry of both CNS and systemic tissues, several of which had not been described, add to the understanding of the stored material in different cell types.

We report a unique case of a minute, occult synovial sarcoma of the lung detected intraoperatively during a pneumothorax repair in a 17-year-old boy. No alternative primary site could be detected upon complete body imaging studies and physical examinations. The diagnosis was confirmed by demonstration of the characteristic SYT/SSX gene fusion by reverse transcriptase polymerase chain reaction (RT-PCR) performed upon RNA extracted from the paraffin block of the biopsy. This case demonstrates the utility of this technique in diagnostic pathology.

We report a case of juxtaposed Wilms' tumor (WT) and cystic nephroma (CN) in a 21-month-old girl which gave rise to radiological diagnostic difficulty. Preoperative chemotherapy was given, resulting in marked tumor necrosis but the cystic nephroma remained untouched. Histological examination showed characteristic features of a triphasic WT and a CN; the two lesions were separated by a thick fibrous capsule. While everybody agrees that WT and cystic partially differentiated nephroblastoma (CPDN) are closely related, there are two opposite views about their relationship to CN. One is that CN may represent the final step in maturation of WT and CPDN. Other authors argue that there is no evidence to support this theory but believe CN might have something in common with nephrogenic rests. We suggest that the two lesions in the present case may have originated from two intralobar nephrogenic rests, which would strengthen the latter view.

A pregnancy with one normal female fetus and a placenta that was divided into halves, one normal the other molar, is described. Genetic analysis shows the molar component to be hyperdiploid/tetraploid but having an identical DNA composition as the normal twin. Because there was no trophoblastic proliferation and the hyperdiploid cells were confined to the villous stroma, and because the molar component was still being perfused by diploid vessels from the normal twin, we believe the mole is derived from polyploidization of the mesenchymal epiblast in a monozygotic twin pregnancy.

This case provides a complete light microscopic, immunophenotypic, and molecular genetic analysis of autoimmune lymphoproliferative syndrome (ALPS), a rare benign cause of dramatic lymphadenopathy with atypical histology and phenotype that may be mistaken for malignancy. The patient is 3-year-old child who was first clinically evaluated at the age of 16 months because of marked generalized lymphadenopathy and hepatosplenomegaly Histologic sections of a cervical lymph node demonstrated a marked paracortical proliferation of occasional small and intermediate-sized lymphocytes and numerous large immunoblasts, the majority of which displayed a CD3⁺, CD43⁺, CD45RO^- (OPD4, UCHL1) CD4^-, CD8^- phenotype on paraffin sections, and which had a CD2⁺, CD3⁺, CD5⁺, CD56^-, Tδ1^-, [CD4^-, CD8^-] double negative profile on flow cytometric analysis. Southern blot analysis did not identify a clonal T or B cell population, and sequencing of the fas gene identified a mutation that caused a single amino acid substitution in the intracytoplasmic death domain of this protein. An enriched population of CD45RO-negative naive T cells in the paracortex may explain the atypical histologic and immunophenotypic features of this case. Greater awareness of this heritable lymphoproliferative disorder will facilitate its recognition and minimize the possibility of misdiagnosis.