Neuron-specific enolase in small-cell carcinoma of the lung: the value of combined immunocytochemistry and serum determination.

The value of neuron-specific enolase (NSE) as a marker for small-cell carcinoma of the lung has been the subject of several reports. Taken together, about 70 per cent of patients with small-cell carcinoma had a raised serum concentration of NSE, and in a majority of patients NSE could be detected immunocytochemically in tumor biopsy speciments. This study examined the diagnostic value of combined immunocytochemical detection and serum determination of NSE in 96 unselected patients with small-cell carcinoma. Seventy-one patients had raised serum concentrations and in 69 a positive immunoreaction for NSE was demonstrated in the biopsy cells. However, for 87 (91%) of the patients, both or either of the NSE assays were positive. The combined use of immunocytochemistry and serum determinations thus gave better information on tumor NSE expression than either method alone. When patients with small or mechanically maltreated biopsy specimens were excluded, the accuracy of the combined assays was even higher (47/49, 96%). We therefore conclude that NSE, although not a specific marker for small-cell carcinoma of the lung, is useful as a complement to conventional diagnostic procedures and, when assayed both in biopsy material and in patient sera, tumor NSE expression can be demonstrated with a high precision.

Endodermal sinus tumour in girls--improved survival in metastatic disease.

In girls, endodermal sinus tumours are rare and malignant germinal tumours. They are highly lethal and it has previously been considered in cases with distant metastases at the time of diagnosis, that there is no satisfactory therapy. However, the rapid development during the last years of highly effective chemotherapeutic agents seems to have improved the survival rate. Already there are a few reports of surviving patients with metastatic disease. In our material of five girls with endodermal sinus tumour, one with metastatic disease has been treated according to a new protocol with combination therapy (cis-VAB). Four years after the operation she is alive and well without any sign of recurrence.

S-100 protein and neuron-specific enolase in amniotic fluid as markers of abdominal wall and neural tube defects in the fetus.

The aim of this study was to determine whether there is increased leakage of neuron-specific enolase (NSE) and S-100 protein into amniotic fluid in pregnancies with neural tube defects, since both these proteins are produced by neural tissue, and to compare the value of these substances for detecting such defects with that of the more conventional techniques of alpha-fetoprotein (AFP) and acetylcholinesterase (AChE) gel electrophoresis. Amniotic samples from 25 mid-pregnancies (15-17 weeks' gestation) with neural tube defects (14 with open spina bifida and 11 with anencephaly) and from seven mid-pregnancies with abdominal wall defects were compared with a control material consisting of 80 amniotic fluid samples from 80 consecutive mid-pregnancy amniocenteses, with normal karyotypes and AFP concentrations. All of the above cases of abnormalities were primarily detected through increased AFP levels in the amniotic fluid. Amniotic fluid samples from 13 pregnancies with fetuses with autosomal chromosomal abnormalities and seven amniotic fluid samples contaminated with blood were also included in the investigation. It is concluded from the results that the conventional AFP assay combined with AChE gel electrophoresis is the best method for screening amniotic fluid for neural tube defects and defects of the abdominal wall. Neither NSE nor S-100 assay alone proved to be superior for the detection of these cases in mid-trimester amniotic fluid. The S-100 assay, however, could give additional information in cases where AChE gel electrophoresis is not decisive; for example, in samples contaminated with blood.

S-100 protein and neuron-specific enolase in cerebrospinal fluid and serum: markers of cell damage in human central nervous system.

The development of a radioimmunoassay for S-100 protein is described. This method was used in combination with a recently developed radioimmunoassay for neuron-specific enolase in cerebrospinal fluid and serum from 47 patients with cerebral infarction, transient ischemic attack, intracerebral hemorrhage, subarachnoid hemorrhage, and head injury. In cerebrospinal fluid, increased concentrations of both S-100 and neuron-specific enolase were found after large infarcts, whereas after small infarcts and transient ischemic attacks, only neuron-specific enolase increased. The increased concentrations of S-100 and/or neuron-specific enolase were noted 18 hours to 4 days after cerebral infarction and transient ischemic attacks. Cerebrospinal fluid concentrations of these proteins also reflected the severity of the disease in patients with intracerebral hematoma, subarachnoid hemorrhage, or head injury. Temporal changes in serum S-100 and neuron-specific enolase concentrations reflected the clinical course in 4 patients. In stroke patients, the S-100 and neuron-specific enolase concentrations may reflect the extent of brain damage and could be useful in selecting patients with major stroke for more aggressive treatment during the acute phase.

Immunocytochemical demonstration of neuron-specific enolase for differential diagnosis of retroperitoneal tumours in children.

Six patients with malignant retroperitoneal tumours causing considerable diagnostic difficulties are reported. In a retrospective investigation the presence of neuron-specific enolase (NSE) was analysed immunocytochemically in specimens of these tumours. In all the patients who had finally got the definite diagnosis neuroblastoma, tumour specimens had a strong reaction for NSE whereas all other tumours did not react. An analysis of NSE already at the primary investigation could have led to an earlier definite diagnosis.

Expression of gamma-subunit of enolase, neuron-specific enolase, in human non-neuroendocrine tumors and derived cell lines.

The gamma-subunit of 2-phospho-D-glycerate hydrolyase, E.C. 4.2.1.11 (enolase), neuron-specific enolase (NSE), is present at high concentrations in neurons and neuroendocrine cells and has therefore recently been introduced as a marker for neuroendocrine tumors. By the indirect methods, immunocytochemistry and radioimmunoassay, NSE has been detected also in some nonneuroendocrine tumors, a finding that could reflect technical artifacts or the capacity for NSE expression in nonneuroendocrine tumor cells. This paper reports on the expression of NSE in human neuroendocrine and nonneuroendocrine tumor specimens and in a panel of permanent human cell lines, by using a direct (enzymatic) and an indirect (radioimmunoassay) method for determination of NSE. We detected NSE in all tested tumor specimens and neuroendocrine tumor cell lines and in a majority (21 of 24) of the nonneuroendocrine tumor cell lines. In general, neuroendocrine tumor specimens and derived tumor cell lines contained more NSE than the nonneuroendocrine tumor specimens and cell lines. However, some of the cultured hematopoietic cell lines (T leukemia and Epstein-Barr virus immortalized B lymphoblastoid cell lines) had NSE levels comparable to those found in some neuroblastoma and small-cell lung carcinoma cell lines. We conclude that NSE is not exclusively expressed in neuroendocrine tumor cells.

Immunocytochemical demonstration of neuron-specific enolase (NSE) in human lung cancers.

An anti-serum against human neuron-specific enolase (NSE), raised in sheep, has been characterized and used for immunocytochemical localization of NSE in paraffin sections of normal tissues and lung cancers. Of the small cell carcinomas (SCC), 69 out of 99 (70%) cases stained with the anti-serum. Maltreated biopsies showed a lower frequency of positive staining (19/39), indicating the importance of well-preserved biopsies. There was no clear difference in the staining between the oat cell and intermediate cell type of SCC. A majority (14 out of 21) of the non-SCC:s (large cell, adenocarcinoma and squamous cell carcinoma) were also stained by the anti-serum. Generally, this staining was weak and it could be blocked by preabsorption of the anti-serum by purified NSE. It is concluded that NSE expression, in conjunction with traditional histology, serves as a useful but not exclusive marker for SCC.

Significance of collateral arterial supply to Wilms' tumours.

The presence of collateral arterial supply was examined by angiography in 19 children with Wilms' tumour. Collateral arterial supply was related to tumour size. Ten of 14 tumours displaying collateral circulation were entirely intrarenal at operation, confirmed by histopathology. Angiography in Wilms' tumour is indicated when the results of urography, ultrasonography or computed tomography are equivocal or extrarenal tumour growth is suggested.

Neuron-specific enolase in mucosal endocrine cells and carcinoid tumours of the small intestine: a comparative study with neuron-specific enolase immunocytochemistry and silver stains.

Endocrine cells of human small intestinal mucosa, small intestinal carcinoids and carcinoid liver metastases were stained with an immunocytochemical technique using an antiserum against neuron-specific enolase (NSE), with the argyrophil technique of Grimelius and with the argentaffin technique of Masson. In the normal mucosa, scattered NSE-immunoreactive cells were seen mainly in the deeper parts of the crypts. These cells, as shown in the same sections, corresponded to the argentaffin and/or argyrophil cells indicating that they were of endocrine type. All intestinal carcinoids (16 cases) displayed NSE immunoreactivity. However, this reaction did not correlate on the cellular level with the silver techniques employed. Thus, many tumour cells were NSE immunoreactive but lacked an argentaffin or argyrophil reaction and vice versa. On the light microscopical level the silver techniques reveal the presence of neurohormonal granules in the tumour cells, while the NSE immunoreactivity appears to disclose neuroendocrine differentiation of the tumour cells irrespective of their hormone and granular content. Out of 13 carcinoid liver metastases, eight displayed strong NSE immunoreactivity, three were weakly stained and two were unreactive. Consecutive or the same tumour sections showed an argentaffin and argyrophil reaction in all carcinoid metastases. Since silver staining provides one type of information and NSE immunocytochemistry another, they provide in combination a good discriminator for neuroendocrine tumours.

Neurone specific enolase: a useful diagnostic serum marker for small cell carcinoma of the lung.

Among lung cancers small cell carcinoma is the most sensitive to chemotherapy and radiation. This has emphasised the importance of an accurate diagnosis of this cell type, and the present study examined the use of serum neurone specific enolase (NSE) as a diagnostic marker for small cell carcinoma. NSE was measured in pretreatment sera from 103 patients with small cell carcinoma and in sera from relevant controls, including patients with other lung cancers, non-malignant lung diseases, and healthy adults. Serum NSE concentration was raised (greater than 25 ng/ml) in 72% of patients with small cell carcinoma. Ninety one per cent of patients with extensive disease and 50% of patients with limited disease were serum NSE positive. Patients with extensive disease in general had higher serum NSE concentrations than patients with limited disease. No definite difference in serum NSE positivity could be shown between oat cell and intermediate cell subtypes. Out of 51 patients with other lung cancers, four (8%) had a raised serum concentration, whereas all patients with non-malignant diseases and healthy individuals had normal serum NSE concentrations. Serum NSE determination seems to be a valuable tool for the diagnosis of small cell carcinoma.

12-O-tetradecanoyl-phorbol-13-acetate-induced differentiation of human neuroblastoma cells is not accompanied by an increase in the intracellular concentration of cyclic AMP.

Cultured human SH-SY5Y neuroblastoma cells could be induced to differentiate morphologically and biochemically followed by growth inhibition, by treatment with 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The cells showed a limited differentiation when treated with substances known to increase the intracellular concentration of cyclic AMP. When these substances were combined with TPA, morphological differentiation and growth inhibition of the cells were potentiated. In contrast, these substances inhibited the TPA-induced increase in noradrenaline concentration and the relative activity of neuron-specific enolase. Both the intracellular concentration of cyclic AMP and the cytosolic level of cyclic AMP-binding components were similar in control and TPA-treated cells. It is suggested that cyclic AMP has a limited and non-regulatory role in the initiation of differentiation of SH-SY5Y cells. The effect of cyclic AMP is probably coupled mainly to the polymerization of microtubules, thus enhancing the morphological differentiation of the cells.

Neuron-specific enolase as a marker for intestinal neurons. An immunocytochemical study of the human intestinal tract.

Surgical specimens from various parts of the human intestinal tract as well as suction biopsy specimens, including mucosa and submucosa of the rectum, were fixed in formalin and embedded in paraffin by routine procedures. The distribution of immunoreactive areas indicating the presence of neuron-specific enolase (NSE) was then determined by using a sheep anti-human-NSE antiserum prepared in our laboratory. The immunocytochemical method revealed, in distinct contrast to other tissue components, the cell bodies of ganglion cells in the submucosa (Meissner's plexus) and in the muscle layers (Auerbach's plexus). The nerve bundles of the submucosa, of the muscle layers, and of the subserosal connective tissue were also stained, whereas the thin nerve processes of the mucosa were identified only rarely. The smooth muscle cells were stained weakly, but this reaction did not interfere with the identification of the neurons and their processes. Immunocytochemical demonstration of NSE is obviously a valuable additional method for visualization of the intrinsic intestinal innervation. It might well be that this technique will be of advantage in the diagnosis of pathologic processes, such as those occurring in Hirschsprung's disease and allied conditions.

Cystic ovaries in a pre-term newborn infant.

Oestrogen-producing ovarian cysts in a very pre-term infant led to swelling of the labia majora and of the lower abdominal wall. Following surgical removal of the left ovary the serum concentration of oestradiol decreased temporarily. Less than 2 weeks after surgery several new cysts appeared in the right ovary. Treatment with medroxyprogesterone acetate resulted in disappearance of these cysts. Simultaneously the serum concentration of oestradiol fell to a normal level.

Retinoic acid-induced differentiation of cultured human neuroblastoma cells: a comparison with phorbolester-induced differentiation.

Cultured human SH-SY5Y neuroblastoma cells differentiated in the presence of retinoic acid (RA) or 12-0-tetradecanoyl-phorbol-13-acetate (TPA). In both cases, the cells acquired long cell processes and the cell growth was partially inhibited. Treatment with RA or TPA resulted in an increased neuron-specific enolase activity, relative to the total cellular enolase activity. At the optimal concentration, TPA induced a 200-fold increase in the concentration of noradrenalin, whereas in RA-treated cells the corresponding increase was only fourfold. Cells treated with a combination of RA and TPA were morphologically differentiated and growth inhibited and had a high relative activity of neuron-specific enolase. The increase in the concentration of noradrenalin induced by TPA was inhibited by RA in a concentration-dependent fashion. However, despite this result there seemed to be no general antagonistic effect of RA on the TPA-induced differentiation. The phenotypes of the cells treated by RA, TPA, or the combination of RA and TPA, did, on the other hand, differ from each other. Our results suggest that RA and TPA induce the SH-SY5Y cells to differentiate along different pathways.

Neuron-specific enolase as a marker for neuroblastoma and small-cell carcinoma of the lung.

The use of neuron-specific enolase (NSE, E.C. 4.2.1.11) as a clinical marker for neuroblastoma and small-cell carcinoma of lung (SCCL) is presented. Both tumors have a high content of NSE as demonstrated enzymatically or by immunocytochemistry. Other retroperitoneal tumors in children and other lung tumors had insignificant NSE concentrations. NSE can thus be used in the differential diagnosis of neuroblastoma and SCCL. 73% of patients with SCCL had elevated serum NSE levels. The corresponding figure for patients with other types of lung cancer was 3%. There was a good correlation between serum NSE levels and the clinical course of patients with SCCL.

Purification and characterization of human neuron-specific enolase: radioimmunoassay development.

Human neuron-specific enolase, NSE, (E.C.4.2.1.11), has been purified from the brain. The method includes a salt precipitation and column chromatography on DEAE-Sephacel, Sephacryl S-300, DEAE-Sepharose and Blue-Sepharose. The specific activity of the purified NSE was 51 units/mg protein and the yield was 28%. The molecular weight of human NSE was compared to the bovine form purified by the same procedure. On Sephadex G 150 both human and bovine NSE chromatographed, in the dimeric form, as 77,000 Dalton proteins, while the molecular weight of the monomer was 45,000 Daltons as determined by SDS poly-acrylamide gel electrophoresis. An antiserum, specific for NSE, was raised in rabbits and a radioimmunoassay developed. The contribution of lyzed blood cells to the serum NSE levels in eight healthy donors was determined and correlated with the haemoglobin content. Minimal haemolysis can add 5-10 ng NSE/ ml to the true serum level. For accurate serum NSE determinations, serum samples must be free from haemolysis.

Surgical implications of congenital chloride diarrhoea.

Congenital chloride diarrhoea (CCD) is an inherited inborn error of metabolism. Hydramnios, premature birth, no passage of meconium and a distended abdomen are the typical features of this disease in the neonate. Loss of Cl- from the intestines leads to urine-like diarrhoea, severe dehydration and alkalosis. Without treatment the mortality rate is high. The abdominal distension may erroneously be ascribed to an intestinal obstruction and thereby lead to unnecessary operations and delay of adequate treatment. There is an overrepresentation of intestinal volvulus in reported cases. Three of the five cases of CCD seen at our departments exemplify these surgical implications.

Neuron-specific enolase as a differential marker in a case of retroperitoneal tumour.

In some cases of retroperitoneal tumour, preoperative investigations fail to conclusively differentiate between neuroblastoma and Wilms' tumour. Diagnostic difficulties can be encountered also intraoperatively, and even the histopathological presentation can be equivocal. According to earlier results, neuron-specific enolase (NSE) in tumour extract can be used as a marker for neuronal tissue and thus offers a new diagnostic criterion differing neuroblastoma from e.g. Wilms' tumour. This paper presents a case of Wilms' tumour where a low content of NSE in the tumour extract correctly indicated a non-neuronal tumour, whereas preoperative presentation and primary histopathology suggested a neuroblastoma.

Experience of the Cohen method for ureteral reimplantation in complicated cases.

Thirty-seven infants and children (41 ureters), a majority with complicating factors (low age, neurogenic bladder, duplication, reoperative surgery), were treated with ureteral reimplantation according to Cohen. The results were good in 90% of the cases. Minor deviations from the original technique, i. e. Dexon instead of chromic catgut as suturing material, catheter splinting of ureter in only 40% and short length of submucosal tunnel in some cases, did not seem to have a negative influence on the results. The method, which is easy and safe to perform, is recommended, particularly in complicated cases.