Comprehensive genetic and histopathologic study reveals three types of neuroblastoma tumors.

PURPOSE: To determine the relationship between multiple genetic features, tumor morphology, and prognosis in neuroblastoma. PATIENTS AND METHODS: The genetic alterations and morphologic features that underpin three histopathologic risk classifications were analyzed in 108 neuroblastoma patients. Tumors were subdivided into four groups based on the three most frequent and prognostically significant genetic alterations (17q gain, 1p deletion, and MYCN amplification), and all other genetic, morphologic, and clinical data were analyzed with respect to these groups. RESULTS: Our analyses identify three nonoverlapping tumor types with distinct genetic and morphologic features, defined here as types 1, 2, and 3. Type 1 tumors show none of the three significant genetic alterations and have good prognosis. Both type 2 (17q gain only or 17q gain and 1p del) and type 3 (17q gain, 1p del, and MYCN amplification) tumors progress. However, these tumor types are distinguished clinically by having significantly different median age at diagnosis and median progression-free survival (PFS). Multivariate analysis indicates that 17q gain is the only independent prognostic factor among all genetic, histopathologic, and clinical factors analyzed. Among histopathologic risk systems, the International Neuroblastoma Pathology Classification was the best predictor of PFS. CONCLUSION: Our results indicate that specific combinations of genetic changes in neuroblastoma tumors contribute to distinct morphologic and clinical features. Furthermore, the identification of two genetically and morphologically distinct types of progressing tumors suggests that possibilities for different therapeutic regimens should be investigated.

Solitary rectal ulcer syndrome in children. A report of three cases.

Solitary rectal ulcer syndrome (SRUS) is a rare condition in children. It is well recognised in the adult literature. We report our experience with 3 patients; two boys presented at a very young age (18 and 24 months, respectively) with a clinical picture simulating inflammatory bowel diseases. The other patient was a fifteen-year-old girl presenting with severe rectal stricture as a result of the syndrome, which is the first to be reported in the paediatric literature.

Lymphatic invasion in Spitz nevi.

Forty-nine Spitz nevi occurring in children were reviewed and sampled extensively in order to assess the incidence of vascular invasion. Evidence of vascular invasion was found in seven (14.3%) cases. The endothelium of such vessels was negative on immunoperoxidase staining for Factor VIII-related antigen suggesting the nevus cells to be in lymphatic channels and not blood vessels. No unusual histological or clinical features characterized the group. All patients are alive and well some years after local excision therapy. It is concluded that lymphatic invasion by nevus cells in Spitz nevi is not uncommon and its presence should not tempt the pathologist into a diagnosis of melanoma.

Recognizable inherited syndrome of progressive central nervous system degeneration and generalized intracranial calcification with overlapping phenotype of the syndrome of Aicardi and Goutières.

Five boys and two girls from a large consanguineous British Muslim family of Pakistani origin are described. All presented from infancy to early childhood with progressive moderate to severe developmental delay, postnatal microcephaly, spastic quadriplegia, refractory seizures, and visual handicap. Cerebrospinal fluid (CSF) pleocytosis was present in three children. Neuroimaging with computerized tomography on three boys and a girl showed generalized cortical atrophy, dilatation of the lateral, third, and fourth ventricles, widening of the surface CSF spaces, hypoplasia of the posterior fossa structures, and multiple and solitary calcifications in the cerebral cortex and punctate calcifications involving basal ganglia, cerebellum, and the Sylvian fissure. Histopathological examination of the brain from three boys and one girl confirmed generalized cortical and cerebellar atrophy with widespread calcifications within the cortical grey and white matter, the basal ganglia, the cerebellum, and in some areas along the capillaries. Investigations excluded a possible nongenetic cause. Parental consanguinity favor autosomal recessive inheritance. This appears to be a recognizable syndrome overlapping the syndrome of Aicardi and Goutières (MIM 225750).

Small intestinal mucosal abnormalities in post-perinatal deaths.

Examination of small intestinal mucosa from cases of post-perinatal death in Sheffield between September 1980 and September 1981 showed mucosal changes before death in 18 of 78 cases (20%). There was no significant difference in prevalence between explained and unexplained deaths, nor was there any positive association with viral isolation from the small intestine. The lesion was much more common in males than females and showed a strong association with bottle feeding--no infant wholly breast fed showed an enteropathy. There was a low incidence of symptoms referrable to the gastrointestinal tract among affected infants, and no appreciable evidence of failure to thrive, as reflected by the postmortem body weight, was present. Mucosal changes of the small intestine in cases of sudden infant death syndrome have previously been reported and attributed to heatstroke. Although the finding of similar lesions in infants who died explicably does not appear to support this view, overheating is difficult to exclude as most of the explained deaths with a mucosal lesion occurred at home.

Upper oesophageal gastric heterotopia: a prospective necropsy study in children.

Three hundred specimens of oesophagus obtained at necropsy from infants and children aged from 0 to 14 years, dying from a variety of causes, were examined prospectively for evidence of gastric heterotopia. Gastric heterotopia was observed in 63 (21%) of the whole series, representing a much higher incidence than reported previously. Excluding deaths in the perinatal period, the incidence appeared to be inversely related to age. Heterotopia was restricted to the subcricoid level of the oesophagus and often showed a close association with lymphoid tissue. There was no association with congenital malformation, and heterotopia occurred more commonly in those infants whose deaths were unexplained, although the reason for this association was unclear.

Sudden infant death and cytomegalovirus inclusion disease.

Four infants, apparently thriving and without clinical evidence of disease, died suddenly at ages ranging from 2 to 6 months. Inclusions bearing cells pathognomonic of cytomegalovirus infection were shown microscopically in a small number of extraneural organs. In view of the lack of associated tissue destruction on microscopy and the apparent well being of the infants before death whether the function of these organs had been impaired to any important degree was questionable: such limited disease, consequently, could not have contributed substantially to the cause of death. The brainstem, on the other hand, consistently showed small numbers of glial nodules. Damage to strategically located neurones associated potentially with the organisation of vital function was a possible basis of sudden death. Alternatively, the small number of glial nodules may have represented a residue of previous more severe brainstem disease, which had possibly started while the baby was in the uterus.

Visceral brown fat necrosis in postperinatal mortality.

Fat necrosis was present in 22 of 400 cases of consecutive postperinatal mortalities investigated to assess the presence and pattern of deep fat necrosis. In just over 50% of the cases of fat necrosis the cause of death was categorised as sudden infant death syndrome, which also showed more severe degrees of necrosis. The mechanism of necrosis may be vascular hypoperfusion, possibly related to shock, and brown adipose tissue, on account of its high metabolic activity and rich capillary plexus, may be particularly vulnerable to infarction. The occurrence of fat necrosis in association with other causes of death did not provide any definite clue as to the nature of the alleged shock.

Splenic macrophages in preterm infants: a necropsy study.

AIMS: To study the frequency and nature of histiocytes in the splenic red pulp of infants who died following complicated immaturity/prematurity. METHODS: Twenty four preterm/immature infants were investigated. Frozen sections of formalin fixed splenic tissue were stained with Oil Red O. Paraffin wax sections from the same tissue were conventionally stained with haematoxylin and eosin. Immunohistochemistry was carried out for a number of macrophage markers. The administration of Intralipid was compared with the presence and extent of tissue macrophages. RESULTS: The spleens of 10 infants showed varying degrees of Oil Red O positivity ranging from mild to strong. In all these cases varying numbers of macrophages were confirmed in the splenic parenchyma in ordinary sections. The immunomarkers indicated that the histiocytes belonged to the macrophage phagocytic system. Of the 10 cases with splenic macrophages all had received Intralipid. Of those not receiving Intralipid none showed splenic macrophages. Seven had received Intralipid but did not have splenic macrophages; they had either only received small amounts of Intralipid or Intralipid was discontinued before death. CONCLUSIONS: Splenic macrophages are common at necropsy in immature/preterm infants. The macrophages are most lucidly demonstrated using Oil Red O staining in frozen sections. There is a strong association between the presence of splenic macrophages and Intralipid administration.

Histochemical demonstration of acetylcholinesterase in neuroblastoma.

The presence of acetylcholinesterase in the tumour cells of neuroblastoma has been shown by enzyme histochemistry. For comparison, some other tumours likely to be found in children and commonly presenting histologically as small cell tumours have also been studied. Acetylcholinesterase activity was seen in rhabdomyosarcoma, but, compared with neuroblastoma, the activity was focal and sparse. One Ewing's tumour and a lymphoblastic lymphoma were negative for the enzyme reaction. Some of the ultrastructural features of neuroblastoma are correlated with the presence of this enzyme. Acetylcholinesterase enzyme histochemistry may provide a useful adjunct in the distinction of neuroblastoma from other small cell tumours.

Comparison of beta enolase and myoglobin as histological markers of rhabdomyosarcoma.

A comparative study of beta enolase and myoglobin as markers of muscle differentiation in rhabdomyosarcoma was carried out, using an immunoperoxidase peroxidase antiperoxidase technique. Material from 26 cases of childhood rhabdomyosarcoma was studied and subdivided into embryonal and alveolar types. Positive cytoplasmic staining for beta enolase was seen in 85% of tumours studied (91% alveolar, 79% embryonal), whereas positive staining for myoglobin was detected in only 69% of tumours (82% alveolar, 64% embryonal). beta Enolase and myoglobin are useful in the histological diagnosis of rhabdomyosarcoma, and of the two, beta enolase seems to be the more sensitive.

Vitreous humour and cerebrospinal fluid hypoxanthine concentration as a marker of pre-mortem hypoxia in SIDS.

AIMS: To assess the rate at which premortem hypoxia occurs in sudden infant death syndrome (SIDS) when compared with death in early childhood. METHODS: The hypoxanthine concentration was measured as a marker of premortem hypoxia in vitreous humour and cerebrospinal fluid samples obtained at necropsy from 119 children whose ages ranged from 1 week to 2 years. RESULTS: Increasing interval between death and necropsy was accompanied by an increase in the hypoxanthine concentration of vitreous humour for the first 24 hours, at a rate of 8.3 mumol/l/hour. Thereafter, there was little change with time, and the results wer corrected to 24 hours according to a regression equation. Cerebrospinal fluid concentrations showed no significant change with time following death. Patients were divided into three groups according to the cause of death: SIDS, cardiac or pulmonary disease, and others. Median values for the cerebrospinal fluid hypoxanthine concentrations were not significantly different among the groups and no difference could be shown between the vitreous humour hypoxanthine concentration in cases of SIDS and those children dying from other causes. Patients with established cardiac or pulmonary disease had a significantly reduced vitreous humour hypoxanthine concentration which may have reflected the premortem use of artificial ventilation. CONCLUSIONS: The results of this study do not support the view that pre-mortem hypoxia is a common feature in SIDS when compared with other causes of death.

An investigation of beta enolase as a histological marker of rhabdomyosarcoma.

Sections from 21 tumours diagnosed as primary or metastatic rhabdomyosarcoma were stained for alpha and beta enolase. The cases were subdivided into embryonal and alveolar subtypes (38% and 62%, respectively). Positive cytoplasmic staining for alpha enolase was seen in all but one case, and cytoplasmic staining for beta enolase was seen in some cells in 18 of the 21 cases (86% of the total, 88% of the alveolar subgroup, and 85% of the embryonal subgroup). No cells stained positively for beta enolase in the control series of neuroblastomas, fibrosarcomas, Wilms' sarcomas, and an osteosarcoma. The results show that beta enolase is a sensitive marker of muscular differentiation in rhabdomyosarcoma.

Small intestinal mucosal fat in childhood enteropathies.

A sequential series of 100 small bowel mucosal biopsies from children was studied to assess the frequency and pattern of mucosal fat staining, and to compare patterns of fat distribution with mucosal structure and clinical diagnosis. Deep mucosal fat was commonly associated with those clinical groups showing normal mucosal structure. While fine granular surface epithelial fat was common in normal and abnormal biopsies, the presence of large fat globules in the surface epithelium was almost entirely limited to biopsies showing villous shortening. Large fat globules in the surface epithelium in coeliac disease and cow's milk sensitive enteropathy were probably related to the more severe degrees of villous abnormality encountered in these clinical groups. However, large fat globules in surface epithelium were also found in a few cases of cow's milk sensitive enteropathy with normal or minimal villous blunting. Fat staining may be a useful additional histological marker to aid in the interpretation of small intestinal mucosal biopsies.

Biochemical and histological assessment of hepatic lipid in sudden infant death syndrome.

A biochemical and histological study of hepatic lipid in children dying from the sudden infant death syndrome (SIDS) and children of a similar age dying explicably are reported. Contrary to a previous report based on histological assessment of hepatic lipid, no significant increase of total lipid content in livers of children dying from SIDS was found. Analysis of hepatic phospholipid fatty acid esters, however, revealed a significant difference between SIDS and children of similar age dying acutely and explicably. The phospholipid abnormality found in SIDS was similar to that found in children dying subacutely with hypoxia and would be consistent with increased cell membrane fluidity. The implications of these findings in the pathogenesis of SIDS are discussed.

Necrosis of the pancreas in the haemolytic uraemic syndrome.

A case of haemolytic uraemic syndrome in a three year old boy is described. The clinical course was complicated by hyperglycaemia, and biochemical assay of plasma showed an almost total lack of pancreatic insulin. Extensive necrosis of the exocrine and endocrine pancreas was found at necropsy. The possible pathological physiology of this complication and its prognostic importance are discussed.

Persistent right umbilical vein--case report and review of literature.

Persistence of the proximal portion of the right umbilical vein is described in a premature infant. Infracardiac total anomalous pulmonary venous connexion and unilateral renal agenesis with ipsilateral phocomelia and unicornuate uterus were associated anomalies. The mother had received carbamazepine therapy for epilepsy. Previously reported cases of persistent proximal right umbilical vein have shown a variety of other malformations but a single umbilical artery has been the most consistent.

Age-related basement membrane thickening of the vocal cords in sudden infant death syndrome (SIDS).

Sudden infant death syndrome (SIDS) remains the leading cause of death in the Western world among infants between the ages of 1 month to 1 year. The diagnosis can only be established at autopsy, at which time no obvious or recognizable pathology is evident. A correlation between thickening of the basement membrane of the true vocal cords and the victim's age was observed in all 51 larynges examined, whereas no such finding was detected in 82 larynges of infants who died of other causes. Thus, the data support the authors' previous morphological results suggesting the use of basement membrane thickening as a marker of positive diagnosis of SIDS at autopsy. The possibility that an immune process may be associated with this syndrome merits further investigation and could prove to be of great importance.

Infant mortality, microglial nodules and parotid CMV-type inclusions.

Nine hundred and fifty-one paediatric autopsies showed 24 cases in which the parotid contained cytomegalovirus-(CMV-) type inclusions. Parotid inclusions were found only during the first two years after birth. Of those with parotid inclusions, 15 showed microglial nodules in the brainstem and/or cerebellum. All the infants with microglial nodules were less than 6 months old. Only four infants without parotid inclusions showed microglial nodules. In just over half of the cases with parotid inclusions and microglial nodules death had been recorded as cases of sudden infant death syndrome. Whether there was any connection between the cause of death and microglial nodules remains uncertain as the microglial nodules were usually sparse and widely distributed. Some theories are proposed in this regard and in connection with those dying with parotid inclusions in the absence of microglial nodules. Until the question is finally resolved it may be prudent to view separately all cases of unexplained sudden death in infancy with evidence of CMV infection.

The possible significance of cytomegaloviral parotitis in infant and early childhood deaths.

OBJECTIVE: To determine whether cytomegaloviral (CMV) parotitis reflects a disseminated disease that increases vulnerability to unexpected death. DESIGN: Necropsy-based cross-sectional study comparing incidences of brain stem microglial nodules and visceral lymphocytic infiltrates in patients with and without CMV parotitis. SUBJECTS: One hundred twelve infants and young children comprising a study group of 40 individuals with CMV parotitis (including 32 whose deaths remained unexplained) and two comparison groups comprising 40 explained and 32 unexplained deaths. MAIN OUTCOME MEASURES: Incidence and variation with age of brain stem microglial nodules and lymphocytic infiltrates in liver and kidneys. RESULTS: Brain stem microglial nodules and lymphocytic infiltrates in liver and kidneys are strongly associated with CMV parotitis; their incidence diminishes with increasing age. CONCLUSIONS: Disseminated disease frequently accompanies CMV parotitis in infants. The resolution of brain stem microglial nodules precedes that of parotitis. Active and previous brain stem involvement may increase vulnerability to unexpected death.